WorldWideScience

Sample records for chondrocyte grafts four-year

  1. Cartilage storage at 4 °C with regular culture medium replacement benefits chondrocyte viability of osteochondral grafts in vitro.

    Science.gov (United States)

    Qi, Jianhong; Hu, Zunjie; Song, Hongqiang; Chen, Bin; Xie, Di; Zhou, Lu; Zhang, Yanming

    2016-09-01

    Maintenance of articular cartilage allografts in culture media is a common method of tissue storage; however, the technical parameters of graft storage remain controversial. In this study, we examined the optimal temperature and culture medium exchange rate for the storage of osteochondral allografts in vitro. Cylindrical osteochondral grafts (n = 120), harvested from the talar joint surface of ten Boer goats, were randomly classified into four groups and stored under the following conditions: Group A1 was maintained at 4 °C in culture medium that was refreshed every 2 days; Group A2 was maintained at 4 °C in the same culture medium, without refreshing; Group B1, was maintained at 37 °C in culture medium that was refreshed every 2 days; Group B2, was maintained at 37 °C in the same culture medium, without refreshing. Chondrocyte viability in the grafts was determined by ethidium bromide/fluorescein diacetate staining on days 7, 21, and 35. Proteoglycan content was measured by Safranin-O staining. Group A1 exhibited the highest chondrocyte survival rates of 90.88 %, 88.31 % and 78.69 % on days 7, 21, and 35, respectively. Safranin O staining revealed no significant differences between groups on days 21 and 35. These results suggest that storage of osteochondral grafts at 4 °C with regular culture medium replacement should be highly suitable for clinical application.

  2. Evaluation and analysis of graft hypertrophy by means of arthroscopy, biochemical MRI and osteochondral biopsies in a patient following autologous chondrocyte implantation for treatment of a full-thickness-cartilage defect of the knee.

    Science.gov (United States)

    Niemeyer, Philipp; Uhl, Markus; Salzmann, Gian M; Morscheid, Yannik P; Südkamp, Norbert P; Madry, Henning

    2015-06-01

    Graft hypertrophy represents a characteristic complication following autologous chondrocyte implantation (ACI) for treatment of cartilage defects. Although some epidemiological data suggest that incidence is associated with first-generation ACI using autologous chondrocyte implantation, it has also been reported in other technical modifications of ACI using different biomaterials. Nevertheless, it has not been described in autologous, non-periosteum, implant-free associated ACI. In addition, little is known about histological and T2-relaxation appearance of graft hypertrophy. The present case report provides a rare case of extensive graft hypertrophy following ACI using an autologous spheres technique with clinical progression over time. Detailed clinical, MR tomographic and histological evaluation has been performed, which demonstrates a high quality of repair tissue within the hypertrophic as well as non-hypertrophic transplanted areas of the repair tissue. No expression of collagen type X (a sign of chondrocyte hypertrophy), only slight changes of the subchondral bone and a nearly normal cell-matrix ratio suggest that tissue within the hypertrophic area does not significantly differ from intact and high-quality repair tissue and therefore seems not to cause graft hypertrophy. This is in contrast to the assumption that histological hypertrophy might cause or contribute to an overwhelming growth of the repair tissue within the transplantation site. Data presented in this manuscript might contribute to further explain the etiology of graft hypertrophy following ACI.

  3. Astrobites: Four Years of Astro-blogging

    Science.gov (United States)

    Faesi, Christopher; Newton, Elisabeth R.; Drout, Maria; Rawls, Meredith L.; Montet, Benjamin; Sanders, Nathan; Astrobites Collaboration

    2015-01-01

    Astrobites (astrobites.com) is now entering its fifth year publishing readable and accessible blog posts summarizing recent research in astrophysics and serving as an online hub for science communication. We continue to focus on our primary goal of making active research accessible to undergraduates by helping to break down technical descriptions and define jargon. However, our readership includes not only undergraduates, but also graduate students and professionals within astronomy, astronomy enthusiasts, and educators. With an average of more than 1000 page views per day, Astrobites reaches a large and diverse audience. We provide not only paper summaries, but also career guidance, practical 'how-to' articles, reports on attending conferences (such as this one), and astronomy news. Astrobites continues to be authored, operated, and led exclusively by a graduate students in astronomy, with our author list spanning the United States and beyond. We reflect on the first four years of Astrobites, including our emphasis on providing both graduate students and undergraduates with opportunities to write about their research for broad audiences, and present results from our latest readership survey. We also look forward at our continued involvement in broader science communication efforts, including ComSciCon (comscicon.com), the series of science communication workshops for graduate students.

  4. Regulative mechanisms of chondrocyte adhesion

    DEFF Research Database (Denmark)

    Schmal, Hagen; Mehlhorn, Alexander T; Fehrenbach, Miriam;

    2006-01-01

    -matrix interaction, as well as collagen type II expression in the cartilage graft after two weeks of in vitro cultivation. Basic fibroblast growth factor (bFGF) treated chondrocytes showed increased adhesion to collagen types I and II, fibronectin, and fibrinogen. Attachment to these investigated proteins......%) increased. A cartilage construct was developed based on a clinically established collagen type I scaffold. In this matrix, more than 80% of the cells could be immobilized by mechanisms of adhesion, filtration, and cell entrapment. Confocal laser microscopy revealed focal adhesion sites as points of cell...... significantly enhanced cell proliferation. Matrix design in cartilage engineering must meet the biological demands of amplified cells, because adhesion of chondrocytes depends on their differentiation status and is regulated by bFGF....

  5. Four years in / four years out : two university buildings 2000-2008

    Energy Technology Data Exchange (ETDEWEB)

    Lovell, J. [Royal Inst. of British Architects, London (United Kingdom); Washington Univ., St. Louis, MO (United States). Graduate School of Architecture and Urban Design

    2009-07-01

    The University of Cambridge commissioned London-based Allies and Morrison to develop and design 2 new buildings for its Sidgwick campus, England. Two naturally ventilated buildings were completed and occupied in 2004. The Estate Management and Building Service at the University of Cambridge is now reviewing the actual energy performance of both buildings in terms of user comfort. The finding will be compared against original projections of the design team. The review will also compare actual running costs against projected costs and feedback from building users and facilities management. The approach taken by client and design team towards the life-time energy performance of the building was examined in terms of comfort and control and load management. A more complex system of sub-metering is now proposed for all new buildings. Post occupancy feedback data was used to evaluate the successes and failures of the project teams in terms of energy performance, thermal comfort and building management communication four years after completion. This paper noted that if viewed over 30 years, the initial construction budget of a building accounts for only a fraction of the true cost of a building. During its lifetime, primary expenditure will likely to be upon personnel costs; work performance and salaries, staff retention and absenteeism. Therefore, the success of a building must be seen in the context of its whole life and the comfort of its users. 14 refs., 9 figs.

  6. Motor Proficiency Predicts Cognitive Ability in Four-Year-Olds

    Science.gov (United States)

    Hernandez, Amanda Martinez; Caçola, Priscila

    2015-01-01

    Research has shown links between motor proficiency and cognition in school-age children, however, few have explored earlier ages. We aimed to determine the association between motor proficiency and cognitive ability in four-year-olds. Motor and cognitive skills were examined in 32 (15 males, 17 females) four-year-olds (±5.59 months) using the…

  7. PetroChina Sees Gas Pipeline Profit in Four Years

    Institute of Scientific and Technical Information of China (English)

    Xiao Lu

    2002-01-01

    @@ Sebei-Xining- Lanzhou pipeline put into operation PetroChina, the nation's largest gas producer,recently said that the US$280-million gas pipeline in Northwest China would generate profits within four years.

  8. Laterality and Motor Skills in Four-Year-Olds.

    Science.gov (United States)

    Tan, Lesley E.

    1985-01-01

    Compared four-year-old left-handed children and children lacking definite hand preference with right-handers on motor skills. Found no differences between left-handers and right-handers of either sex, but the children lacking hand preference had lower scores. Possible sex differences and implications for the education of children lacking…

  9. The Hidden Costs of Low Four-Year Graduation Rates

    Science.gov (United States)

    Sullivan, Daniel F.

    2010-01-01

    The single most important step colleges and universities--especially public colleges and universities--can take to lower the student and family cost of college attendance is to improve retention, thereby increasing the four-year graduation rate. The author believes that institutions with high rates of retention to graduation have those high rates…

  10. Four Years of Portability: Perspectives on a Laptop Program.

    Science.gov (United States)

    Barrett, Joanne

    2002-01-01

    Reviews a laptop computer technology program that has been in effect for nearly four years in a middle school in Bethesda, Maryland. Topics include initial opposition from parents; benefits to teachers, students, and learning; copyright issues; problems with plagiarism; accessibility; accountability; and email issues. (LRW)

  11. Partnerships Between TYCs and Four-Year Institutions

    Science.gov (United States)

    Clay, Keith; Sabella, Mel; Price, Edward

    2012-02-01

    We are faced with a severe shortage of certificated STEM teachers, especially in physics. At the present time, four year institutions are struggling to address this shortage with only a handful of schools graduating two or more physics educators each year. The science education community must therefore focus more attention on the important role of the two year college (TYC), where one third to one half of our STEM majors begin their college careers. In this session, we will explore the obstacles and prospects for recruiting math and science teachers from TYCs from the perspectives of three institutions who are interested in developing meaningful collaborations between the TYC and the four year institution. Group discussions will consider options and look for solutions.

  12. Video-EEG recording: a four-year clinical audit.

    LENUS (Irish Health Repository)

    O'Rourke, K

    2012-02-03

    In the setting of a regional neurological unit without an epilepsy surgery service as in our case, video-EEG telemetry is undertaken for three main reasons; to investigate whether frequent paroxysmal events represent seizures when there is clinical doubt, to attempt anatomical localization of partial seizures when standard EEG is unhelpful, and to attempt to confirm that seizures are non-epileptic when this is suspected. A clinical audit of all telemetry performed over a four-year period was carried out, in order to determine the clinical utility of this aspect of the service and to determine means of improving effectiveness in the unit. Analysis of the data showed a high rate of negative studies with no attacks recorded. Of the positive studies approximately 50% showed non-epileptic attacks. Strategies for improving the rate of positive investigations are discussed.

  13. Analyzing the Bitcoin Network: The First Four Years

    Directory of Open Access Journals (Sweden)

    Matthias Lischke

    2016-03-01

    Full Text Available In this explorative study, we examine the economy and transaction network of the decentralized digital currency Bitcoin during the first four years of its existence. The objective is to develop insights into the evolution of the Bitcoin economy during this period. For this, we establish and analyze a novel integrated dataset that enriches data from the Bitcoin blockchain with off-network data such as business categories and geo-locations. Our analyses reveal the major Bitcoin businesses and markets. Our results also give insights on the business distribution by countries and how businesses evolve over time. We also show that there is a gambling network that features many very small transactions. Furthermore, regional differences in the adoption and business distribution could be found. In the network analysis, the small world phenomenon is investigated and confirmed for several subgraphs of the Bitcoin network.

  14. The Romanian economy - four years after the EU accession

    Directory of Open Access Journals (Sweden)

    Laurian LUNGU

    2011-12-01

    Full Text Available The first four years of Romania’s membership confirmed that the accession to the EU had, overall, positive effects on the economy. Although the pace of structural reforms had slowed down after 2007, they picked up again after the start of financial crisis, at the end of 2008. Households’ wealth and purchasing power have increased despite the recent drop over the last two years. Looking ahead, Romania’s economy will need to enhance its competitiveness and pursue a growth model which would make it less dependent on external shocks. Economic strategies leading to both physical and human capital accumulation would need to be clearly defined and supported by all political parties. With monetary policy aimed at maintaining price stability, fiscal policy would have a defining role in preserving and enhancing competitiveness. The pursuance of sound fiscal and monetary policies would be paramount in achieving sustained growth in the years to come.

  15. Postural changes in dental hygienists. Four-year longitudinal study.

    Science.gov (United States)

    Barry, R M; Woodall, W R; Mahan, J M

    1992-01-01

    Numerous surveys identify the occurrence of musculoskeletal complaints as a concern in dentistry. However, no longitudinal data exist to indicate whether postural changes occur as a result of practicing dental hygiene. The purpose of this preliminary, four-year longitudinal study was to investigate whether any postural changes developed during the hygienists' clinical education and/or during subsequent dental hygiene practice after one and/or two years. It was anticipated that the awkward positions and intense physical demands placed on hygienists might initiate musculoskeletal problems, but that no postural changes would occur over this short period of time. Nine of 10 dental hygienists in the graduating class of 1987 were surveyed for existing musculoskeletal complaints, and the subjects were photographed for a measurement of postural change. Responses from participants indicated an increase in musculoskeletal-related complaints in each of the six areas investigated. The photographic findings indicated that one of the nine hygienists showed an increase in forward head posture, a postural change.

  16. [The health of industrial employees four years after compulsory redundancy].

    Science.gov (United States)

    Beale, N; Nethercott, S

    1987-09-01

    A controlled, longitudinal study of the health of workers made redundant when a meat products factory closed has been performed using morbidity data extracted from the records of a group general practice. Increases in consultation rates and the number of visits to hospital outpatient departments in the group made redundant are contrasted with opposite trends in a control group who remained securely employed. As in earlier findings, the increases in morbidity in the study group began when they learned that their jobs were in jeopardy.The subsequent employment history of those made redundant was obtained by questionnaire. In the four years after redundancy, 50 of the 76 men in-the study group found new full-time jobs. The other 26 men remained out of work for most of this time or were made redundant once again. This 'jobless' group consulted their general practitioners 57% more often about 13% more illnesses, were referred to hospital outpatient departments 63% more often and visited hospital 208% more frequently than when enjoying secure employment. During an intervening two-year period of job insecurity, there were increases of 45%, 9%, 25% and 28% respectively, for this jobless group.The implications of these findings for primary care, for the National Health Service and for future research are discussed in the present context of high levels of unemployment.

  17. Four years of Ulysses dust data 1996 to 1999

    CERN Document Server

    Krüger, H; Landgraf, M; Dermott, S; Fechtig, H; Gustafson, B A; Hamilton, D P; Hanner, M S; Horányi, M; Kissel, J; Lindblad, B A; Linkert, D; Linkert, G; Mann, I; McDonnell, J A M; Morfill, G E; Polanskey, C; Schwehm, G; Srama, R A; Zook, H A

    2001-01-01

    The Ulysses spacecraft is orbiting the Sun on a highly inclined ellipse ($ i = 79^{\\circ}$, perihelion distance 1.3 AU, aphelion distance 5.4 AU). Between January 1996 and December 1999 the spacecraft was beyond 3 AU from the Sun and crossed the ecliptic plane at aphelion in May 1998. In this four-year period 218 dust impacts were recorded with the dust detector on board. We publish and analyse the complete data set of both raw and reduced data for particles with masses $\\rm 10^{-16} g$ to $\\rm 10^{-8}$ g. Together with 1477 dust impacts recorded between launch of Ulysses and the end of 1995 published earlier \\cite{gruen1995c,krueger1999b}, a data set of 1695 dust impacts detected with the Ulysses sensor between October 1990 and December 1999 is now available. The impact rate measured between 1996 and 1999 was relatively constant with about 0.2 impacts per day. The impact direction of the majority of the impacts is compatible with particles of interstellar origin, the rest are most likely interplanetary parti...

  18. FACT. More than four years of blazar monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Dorner, Daniela [Universitaet Wuerzburg (Germany); Collaboration: FACT-Collaboration

    2016-07-01

    Since October 2011, the First G-APD Cherenkov Telescope (FACT) has been collecting more than 5500 hours of physics data. Thanks to the silicon based photosensors (SiPMs, aka G-APDs), observations during bright ambient light like full moon can be carried out without degradation of the sensors. Keeping the gain of the SiPMs stable using an online feedback system, a stable and homogeneous detector performance is achieved. Based on this and an automatic data taking procedure, an unbiased longterm data sample is collected. An automatic quick look analysis provides results shortly after the data are taken allowing to send flare alerts within the same night. The main targets for FACT are the bright TeV blazars Mrk 421 and Mrk 501 which are monitored since January 2012. In addition, several other sources like for example the Crab Nebula, 1ES 1959+650, 1ES 2344+54.1 are observed. In this presentation, the results from more than four years of monitoring are summarized. Several flares from Mrk 501 and Mrk 421 are discussed in the multi-wavelength (MWL) context. Mrk 501 underwent major outbursts in June 2012 and June 2014 during the yearly MWL campaigns. Mrk 421 showed a bright flare in April 2013 where also MWL observations are available. 1ES 1959+650 showed enhanced flux in autumn 2015. Results from these observations are discussed.

  19. Survey on the Transferability of Associate's Degree to Four-Year Institutions.

    Science.gov (United States)

    Rubi, David C.

    In January 1994, the Arizona State Board of Directors for Community Colleges (ASBDCC) conducted a survey of state four-year systems nationwide to determine the existence of standards regarding the transferability of associate degrees and associate degree students' academic standing at receiving four-year institutions. Questionnaires were mailed to…

  20. The Cost of College Attrition at Four-Year Colleges & Universities. Policy Perspectives

    Science.gov (United States)

    Raisman, Neal

    2013-01-01

    This report, an analysis of 1669 U.S. institutions, is a first-time study of the relationship of attrition to revenues lost in four-year public, private, and for-profit colleges and universities on an annual basis. The report is the result of a study investigating the financial impact of attrition on four-year colleges and universities. Other…

  1. Customized biomaterials to augment chondrocyte gene therapy.

    Science.gov (United States)

    Aguilar, Izath Nizeet; Trippel, Stephen; Shi, Shuiliang; Bonassar, Lawrence J

    2017-02-07

    A persistent challenge in enhancing gene therapy is the transient availability of the target gene product. This is particularly true in tissue engineering applications. The transient exposure of cells to the product could be insufficient to promote tissue regeneration. Here we report the development of a new material engineered to have a high affinity for a therapeutic gene product. We focus on insulin-like growth factor-I (IGF-I) for its highly anabolic effects on many tissues such as spinal cord, heart, brain and cartilage. One of the ways that tissues store IGF-I is through a group of insulin like growth factor binding proteins (IGFBPs), such as IGFBP-5. We grafted the IGF-I binding peptide sequence from IGFBP-5 onto alginate in order to retain the endogenous IGF-I produced by transfected chondrocytes. This novel material bound IGF-I and released the growth factor for at least 30days in culture. We found that this binding enhanced the biosynthesis of transfected cells up to 19-fold. These data demonstrate the coordinated engineering of cell behavior and material chemistry to greatly enhance extracellular matrix synthesis and tissue assembly, and can serve as a template for the enhanced performance of other therapeutic proteins.

  2. Evolution of Autologous Chondrocyte Repair and Comparison to Other Cartilage Repair Techniques

    Directory of Open Access Journals (Sweden)

    Ashvin K. Dewan

    2014-01-01

    Full Text Available Articular cartilage defects have been addressed using microfracture, abrasion chondroplasty, or osteochondral grafting, but these strategies do not generate tissue that adequately recapitulates native cartilage. During the past 25 years, promising new strategies using assorted scaffolds and cell sources to induce chondrocyte expansion have emerged. We reviewed the evolution of autologous chondrocyte implantation and compared it to other cartilage repair techniques. Methods. We searched PubMed from 1949 to 2014 for the keywords “autologous chondrocyte implantation” (ACI and “cartilage repair” in clinical trials, meta-analyses, and review articles. We analyzed these articles, their bibliographies, our experience, and cartilage regeneration textbooks. Results. Microfracture, abrasion chondroplasty, osteochondral grafting, ACI, and autologous matrix-induced chondrogenesis are distinguishable by cell source (including chondrocytes and stem cells and associated scaffolds (natural or synthetic, hydrogels or membranes. ACI seems to be as good as, if not better than, microfracture for repairing large chondral defects in a young patient’s knee as evaluated by multiple clinical indices and the quality of regenerated tissue. Conclusion. Although there is not enough evidence to determine the best repair technique, ACI is the most established cell-based treatment for full-thickness chondral defects in young patients.

  3. Chondrocyte physiopathology and drug efficacy.

    Science.gov (United States)

    Serni, U; Mannoni, A

    1991-01-01

    After a brief exposition on the physiopathology of cartilage, and characteristic features of chondrocytes and proteoglycans (PGs) in osteoarthritis (OA), it is underlined how different molecules of GAGs and aggregated PGs added to the culture media can prevent damage and reduction of GAGs and fibril production in chondrocytes cultured with NSAIDs and corticosteroids. In animal models of OA, the local or general administration of GAGPS reduces the proteinase activity, the level of uronic acid in synovial fluid and the number of inflammatory cells in synovia. In the Pond-Nuki dog, GAGPS improves the cartilage surface. These favourable events can also occur in human OA, where it is, moreover, difficult to monitor the patients. For this purpose, patients must be selected in the first two stages of primary OA, and followed using NMR, the only device able to scan cartilage and subchondral bone, to determine their consistency and thickness, and to provide information on water content.

  4. Articular chondrocyte metabolism and osteoarthritis

    Energy Technology Data Exchange (ETDEWEB)

    Leipold, H.R.

    1989-01-01

    The three main objectives of this study were: (1) to determine if depletion of proteoglycans from the cartilage matrix that occurs during osteoarthritis causes a measurable increase of cartilage proteoglycan components in the synovial fluid and sera, (2) to observe what effect intracellular cAMP has on the expression of matrix components by chondrocytes, and (3) to determine if freshly isolated chondrocytes contain detectable levels of mRNA for fibronectin. Canine serum keratan sulfate and hyaluronate were measured to determine if there was an elevation of these serum glycosaminoglycans in a canine model of osteoarthritis. A single intra-articular injection of chymopapain into a shoulder joint increased serum keratan sulfate 10 fold and hyaluronate less than 2 fold in 24 hours. Keratan sulfate concentrations in synovial fluids of dogs about one year old were unrelated to the presence of spontaneous cartilage degeneration in the joints. High keratan sulfate in synovial fluids correlated with higher keratan sulfate in serum. The mean keratan sulfate concentration in sera of older dogs with osteoarthritis was 37% higher than disease-free controls, but the difference between the groups was not statistically significant. Treatment of chondrocytes with 0.5 millimolar (mM) dibutyryl cAMP (DBcAMP) caused the cells to adopt a more rounded morphology. There was no difference between the amount of proteins synthesized by cultures treated with DBcAMP and controls. The amount of fibronectin (FN) in the media of DBcAMP treated cultures detected by an ELISA was specifically reduced, and the amount of {sup 35}S-FN purified by gelatin affinity chromatography decreased. Moreover, the percentage of FN containing the extra domain. A sequence was reduced. Concomitant with the decrease in FN there was an increase in the concentration of keratan sulfate.

  5. A spike at benefit exhaustion: Still possible after four years of unemployment

    DEFF Research Database (Denmark)

    Lyk-Jensen, Stéphanie; Weatherall, Cecilie Dohlmann

    European policymakers wonder whether the long-term unemployed can still find jobs if the benefit period is shortened. We investigate this question by analysing how long-term unemployed react to the threat of running out of unemployment insurance (UI) after receiving it for almost four years....... The empirical analysis is based on very precise administrative records of men’s unemployment spells. To identify the effect of UI exhaustion, we exploit a legislative change in the duration of benefits that progressively reduced UI entitlement from five to four years. Our results show a spike 3-6 months before...... benefit exhaustion for long-term unemployed....

  6. Librarian integration in a four-year medical school curriculum: a timeline.

    Science.gov (United States)

    MacEachern, Mark; Townsend, Whitney; Young, Kristen; Rana, Gurpreet

    2012-01-01

    The Taubman Health Sciences Library (THL) is integrated in all four years of the University of Michigan Medical School (UMMS) curriculum. Information resources are introduced at strategic points throughout the curriculum so that students receive training at times when they are most likely to need the resource. Most of the core instruction sessions are taught in teams that consist of librarians and UMMS faculty, which provides unique learning opportunities for students. This article describes each THL instruction activity in the four-year undergraduate UMMS curriculum and provides commentary on the overall effectiveness of this integrated approach to instruction.

  7. Posttraumatic Chondrocyte Apoptosis in the Murine Xiphoid

    Science.gov (United States)

    Davis, Christopher G.; Eisner, Eric; McGlynn, Margaret; Shelton, John M.; Richardson, James

    2013-01-01

    Objective. To demonstrate posttraumatic chondrocyte apoptosis in the murine xiphoid after a crush-type injury and to ultimately determine the pathway (i.e., intrinsic or extrinsic) by which chondrocytes undergo apoptosis in response to mechanical injury. Design. The xiphoids of adult female wild-type mice were injured with the use of a modified Kelly clamp. Postinjury xiphoid cartilage was analyzed via 3 well-described independent means of assessing apoptosis in chondrocytes: hematoxylin and eosin staining, terminal deoxynucleotidyl transferase dUTP nick end labeling assay, and activated caspase-3 staining. Results. Injured specimens contained many chondrocytes with evidence of apoptosis, which is characterized by cell shrinkage, chromatin condensation, nuclear fragmentation, and the liberation of apoptotic bodies. There was a statistically significant increase in the number of chondrocytes undergoing apoptosis in the injured specimens as compared with the uninjured specimens. Conclusions. Chondrocytes can be stimulated to undergo apoptosis as a result of mechanical injury. These experiments involving predominantly cartilaginous murine xiphoid in vivo establish a baseline for future investigations that employ the genetic and therapeutic modulation of chondrocyte apoptosis in response to mechanical injury. PMID:26069679

  8. Transduction of anti-cell death protein FNK suppresses graft degeneration after autologous cylindrical osteochondral transplantation.

    Science.gov (United States)

    Nakachi, Noriki; Asoh, Sadamitsu; Watanabe, Nobuyoshi; Mori, Takashi; Matsushita, Takashi; Takai, Shinro; Ohta, Shigeo

    2009-03-01

    This study shows that artificial super antiapoptotic FNK protein fused with a protein transduction domain (PTD-FNK) maintains the quality of osteochondral transplant by preventing chondrocyte death. Cylindrical osteochondral grafts were obtained from enhanced green fluorescent protein (EGFP)-expressing transgenic rats, in which living chondrocytes express green fluorescence, and submerged into medium containing PTD-FNK, followed by transplantation into cartilage defects of wild-type rats by impact insertion simulating autologous transplantation. The tissues were histologically evaluated by hematoxylin-eosin and Safranin-O staining. At 1 week, chondrocyte alignment was normal in the PTD-FNK treatment group, whereas all grafts without PTD-FNK treatment showed mixed cluster cell distribution. At 4 weeks, all grafts with PTD-FNK treatment showed almost normal matrix, whereas two grafts without PTD-FNK treatment showed fibrocartilage. Notably, all grafts with PTD-FNK retained high intensity of Safranin-O staining, but all grafts without PTD-FNK largely lost Safranin-O staining. PTD-FNK significantly suppressed a decrease in the survival rate and the density of EGFP-positive cells at 1 and 2 weeks, and this tendency continued at 4 weeks. The results of terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-nick end-labeling staining showed that PTD-FNK inhibited cell death, indicating that PTD-FNK protects chondrocyte death and suppresses graft degeneration.

  9. Four-Year Myth: Make College More Affordable. Restore the Promise of Graduating on Time

    Science.gov (United States)

    Complete College America, 2014

    2014-01-01

    In American higher education, it has become the accepted standard to measure graduation rates at four-year colleges on a six-year time frame. Evaluations of two-year community colleges are now based on three-year graduation rates. Metrics like these are unacceptable, especially when we consider that students and their families are trying…

  10. Building a Four Year Post-Secondary Curriculum in Educational Science and Technology.

    Science.gov (United States)

    Plomp, Tjeerd; Verhagen, Plon

    This report begins by providing background information on educational technology in the Netherlands, differences between a four-year program in the Netherlands and undergraduate programs in the United States, and the structure of Dutch university programs. The need for a Department of Educational Science and Technology at the Twente University of…

  11. The Transfer Playbook: Essential Practices for Two- and Four-Year Colleges

    Science.gov (United States)

    Wyner, Joshua; Deane, K. C.; Jenkins, Davis; Fink, John

    2016-01-01

    With their "open door" admissions policies and lower tuition, community colleges offer a gateway to higher education for millions of American students. Indeed, students who enter higher education through community colleges are much more likely than those who start at four-year institutions to be low-income or the first in their family to…

  12. Is Articulation from Two- to Four-Year Colleges an Allowed or a Forbidden Transition?

    Science.gov (United States)

    Susskind, Tamar Y.

    1997-01-01

    Argues that patterns of student flow between two- and four-year institutions are not linear and that the permutations of student situations affect the datasets that are used to both defend and question the effectiveness and transferability of two-year programs. (DDR)

  13. Swim Test Requirements at Four-Year Universities in the United States.

    Science.gov (United States)

    Ormond, Frank; And Others

    1997-01-01

    Surveys of 331 four-year universities nationwide examined swim test requirements and reasons for administering or not administering swim tests. Responses from 331 returned questionnaires indicated that 5% of the schools had swim test requirements, though 25% had them previously. Lack of a physical education requirement was the most commonly cited…

  14. Development of health over four years among middle-aged and older Europeans

    DEFF Research Database (Denmark)

    Vestergaard, Sonja; Eriksen, Mette Lindholm; Andersen-Ranberg, Karen

    2013-01-01

    Older adults in Eastern and Central European countries have a higher risk of developing poor self-rated health over four years. The same geographical pattern is seen for a higher risk of developing hypertension and diabetes. Low educational level significantly increases the risk of developing poor...

  15. Enhancing Software Engineering Education through Open Source Projects: Four Years of Students' Perspectives

    NARCIS (Netherlands)

    Papadopoulos, P.M.; Stamelos, I.G.; Meiszner, A.

    2015-01-01

    This paper presents the results after four years of running of an instructional method that utilizes free/libre open source software (FLOSS) projects as tools for teaching software engineering in formal education. In the last four academic years, a total of 408 juniors majoring in Informatics (in a

  16. Perceptions and Meanings Constructed by Participants in a Four-Year Instructional Coaching Project

    Science.gov (United States)

    Perry Hummons, Monica

    2012-01-01

    This qualitative study was conducted in the Midwestern U.S. It explored the stories of 27 educators who participated in a four-year instructional coaching project in a large urban school district. Subjects shared their perceptions of and meanings they constructed as a result of their experiences in the coaching project through participation in a…

  17. Accelerating the transition to employment at benefit exhaustion: Still possible after four years of unemployment?

    DEFF Research Database (Denmark)

    Vincent, Stéphanie; Weatherall, Cecilie Dohlmann; Geerdsen, Lars Pico

    2017-01-01

    This paper provides evidence of how the long-term unemployed react to the threat of running out of unemployment insurance (UI) after receiving it for nearly four years. To identify the effect of UI exhaustion, we make use of a 1999 Danish legislative change that progressively reduced potential be...

  18. Web Page Change and Persistence-A Four-Year Longitudinal Study.

    Science.gov (United States)

    Koehler, Wallace

    2002-01-01

    Discussion of changes in the topography of the Web focuses on changes to an existing set of Web documents over a four-year period. Highlights include the life cycle of Web objects; changes to Web objects; measures of change; Web page demise; and Web page changes, including hypertext links, content change, and structural change. (LRW)

  19. Self-Repair of Speech by Four-Year-Old Finnish Children

    Science.gov (United States)

    Salonen, Tuuli; Laakso, Minna

    2009-01-01

    The aim of this study was to examine what four-year-old children repair in their speech. For this purpose, conversational self-repairs (N = 316) made by two typically developing Finnish-speaking children (aged 4 ; 8 and 4 ; 11) were examined. The data comprised eight hours of natural interactions videotaped at the children's homes. The tapes were…

  20. Reliability of Objective Structured Clinical Examinations: Four Years of Experience in a Surgical Clerkship.

    Science.gov (United States)

    Mann, Karen V.; And Others

    1990-01-01

    Four years of experience with an objective structured clinical examination (OSCE) following an eight-week surgical clerkship (n=356 students) are reported, including data on mean student performance across years, reliability coefficients, and generalizability. Implications for improvement and development of OSCE are discussed. (Author/MSE)

  1. Four-year outcome after early withdrawal of antiepileptic drugs in childhood epilepsy

    NARCIS (Netherlands)

    Geerts, AT; Niermeijer, JMF; Arts, WFM; Brouwer, OF; Stroink, H; Peeters, EAJ; van Donselaar, CA

    2005-01-01

    Four-year follow-up of children with epilepsy included in a randomized trial of early withdrawal of antiepileptic drugs showed that 51% achieved a terminal remission of at least 2 years without medication and 21% with medication; 15% had seizures during the fourth year. Early medication withdrawal i

  2. Efficiency-Based Funding for Public Four-Year Colleges and Universities

    Science.gov (United States)

    Sexton, Thomas R.; Comunale, Christie L.; Gara, Stephen C.

    2012-01-01

    We propose an efficiency-based mechanism for state funding of public colleges and universities using data envelopment analysis. We describe the philosophy and the mathematics that underlie the approach and apply\\break the proposed model to data from 362 U.S. public four-year colleges and universities. The model provides incentives to institution…

  3. Performance of Maryland Community College Transfer Students at Public Four-Year College and Universities

    Science.gov (United States)

    Maryland Higher Education Commission, 2011

    2011-01-01

    This data report provides statistics about the number and percentage of Maryland community college students, who earned a bachelor's degree from a public four-year campus in the state within a specified period of years. These students are: (1) Community college students who transferred during the 2004-2005 academic year who received a bachelor's…

  4. The Fermi GBM Gamma-Ray Burst Spectral Catalog: Four Years of Data

    NARCIS (Netherlands)

    Gruber, D.; Goldstein, A.; Weller von Ahlefeld, V.; Bhat, N.P.; Bissaldi, E.; Briggs, M.S.; Byrne, D.; Cleveland, W.H.; Connaughton, V.; Diehl, R.; Fishman, G.J.; Fitzpatrick, G.; Foley, S.; Gibby, M.; Giles, M.M.; Greiner, J.; Guiriec, S.; van der Horst, A.J.; von Kienlin, A.; Kouveliotou, C.; Layden, E.; Lin, L.; Meegan, C.A.; McGlynn, S.; Paciesas, W.S.; Pelassa, V.; Preece, R.D.; Rau, A.; Wilson-Hodge, C.A.; Xiong, S.; Younes, G.; Yu, H-F.

    2014-01-01

    In this catalog we present the updated set of spectral analyses of gamma-ray bursts (GRBs) detected by the Fermi Gamma-Ray Burst Monitor during its first four years of operation. It contains two types of spectra, time-integrated spectral fits and spectral fits at the brightest time bin, from 943 tri

  5. A Four-Year Follow-Up Study of Underachieving College Freshmen

    Science.gov (United States)

    Valine, Warren J.

    1976-01-01

    A four-year follow-up study of underachieving college freshmen measured changes in the self-concept of those still in college and those who had dropped out. Significant differences generally favored college seniors. Findings also indicate that dropping out of college may be a positive experience. (Author)

  6. [Chondrocyte mecanobiology. Application in cartilage tissue engineering].

    Science.gov (United States)

    Stoltz, Jean François; Netter, Patrick; Huselstein, Céline; de Isla, Natalia; Wei Yang, Jing; Muller, Sylvaine

    2005-11-01

    Cartilage is a hydrated connective tissue that withstands and distributes mechanical forces within joints. Chondrocytes utilize mechanical signals to maintain cartilaginous tissue homeostasis. They regulate their metabolic activity through complex biological and biophysical interactions with the extracellular matrix (ECM). Some mechanotransduction mechanisms are known, while many others no doubt remain to be discovered. Various aspects of chondrocyte mechanobiology have been applied to tissue engineering, with the creation of replacement tissue in vitro from bioresorbable or non-bioresorbable scaffolds and harvested cells. The tissues are maintained in a near-physiologic mechanical and biochemical environment. This paper is an overview of both chondrocyte mechanobiology and cartilage tissue engineering

  7. ROCK inhibitor prevents the dedifferentiation of human articular chondrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Emi [Department of Orthopaedic Surgery, Science of Functional Recovery and Reconstruction, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikatacho, Kitaku, Okayama 700-8558 (Japan); Furumatsu, Takayuki, E-mail: matino@md.okayama-u.ac.jp [Department of Orthopaedic Surgery, Science of Functional Recovery and Reconstruction, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikatacho, Kitaku, Okayama 700-8558 (Japan); Kanazawa, Tomoko; Tamura, Masanori; Ozaki, Toshifumi [Department of Orthopaedic Surgery, Science of Functional Recovery and Reconstruction, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 2-5-1 Shikatacho, Kitaku, Okayama 700-8558 (Japan)

    2012-03-30

    Highlights: Black-Right-Pointing-Pointer ROCK inhibitor stimulates chondrogenic gene expression of articular chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor prevents the dedifferentiation of monolayer-cultured chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor enhances the redifferentiation of cultured chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor is useful for preparation of un-dedifferentiated chondrocytes. Black-Right-Pointing-Pointer ROCK inhibitor may be a useful reagent for chondrocyte-based regeneration therapy. -- Abstract: Chondrocytes lose their chondrocytic phenotypes in vitro. The Rho family GTPase ROCK, involved in organizing the actin cytoskeleton, modulates the differentiation status of chondrocytic cells. However, the optimum method to prepare a large number of un-dedifferentiated chondrocytes is still unclear. In this study, we investigated the effect of ROCK inhibitor (ROCKi) on the chondrogenic property of monolayer-cultured articular chondrocytes. Human articular chondrocytes were subcultured in the presence or absence of ROCKi (Y-27632). The expression of chondrocytic marker genes such as SOX9 and COL2A1 was assessed by quantitative real-time PCR analysis. Cellular morphology and viability were evaluated. Chondrogenic redifferentiation potential was examined by a pellet culture procedure. The expression level of SOX9 and COL2A1 was higher in ROCKi-treated chondrocytes than in untreated cells. Chondrocyte morphology varied from a spreading form to a round shape in a ROCKi-dependent manner. In addition, ROCKi treatment stimulated the proliferation of chondrocytes. The deposition of safranin O-stained proteoglycans and type II collagen was highly detected in chondrogenic pellets derived from ROCKi-pretreated chondrocytes. Our results suggest that ROCKi prevents the dedifferentiation of monolayer-cultured chondrocytes, and may be a useful reagent to maintain chondrocytic phenotypes in vitro for chondrocyte

  8. Effect of freezing on rabbit cultured chondrocytes

    Directory of Open Access Journals (Sweden)

    R.R Filgueiras

    2011-02-01

    Full Text Available This work evaluated the effect of freezing on chondrocytes maintained in culture, aiming the establishment of a cell bank for future application as heterologous implant. Chondrocytes extracted from joint cartilage of nine healthy New Zealand White rabbits were cultivated and frozen with the cryoprotector 5% dimethylsulfoxide for six months. Phenotypic and scanning electron microscopy analyses were carried out to identify morphological and functional differences between fresh and thawed cells. After enzymatic digestion, a total of 4.8x10(5cells per rabbit were obtained. Fresh chondrocytes showed a high mitotic rate and abundant matrix was present up to 60 days of culture. Loss of phenotypic stability was notable in the thawed chondrocytes, with a low labeling of proteoglycans and weak immunostaining of type II collagen. The present study showed important loss of chondrocyte viability under the freezing conditions. For future in vivo studies of heterologous implant, these results suggests that a high number of cells should be implanted in the host site in order to achieve an adequate number of viable cells. Furthermore, the chondrocytes should be implanted after two weeks of culture, when the highest viability rate is found

  9. Policy Commentary/Commentaire BC's Carbon Tax Shift Is Working Well after Four Years (Attention Ottawa)

    OpenAIRE

    Stewart Elgie; Jessica McClay

    2013-01-01

    British Columbia's introduction in 2008 of a revenue-neutral carbon tax shift was controversial. This analysis compares changes in fuel consumption, greenhouse gas emissions, and gross domestic product (GDP) between British Columbia and the rest of Canada. It finds that in the four years since the tax was introduced, British Columbia's per capita consumption of fuels subject to the tax has declined by 19 percent compared to the rest of Canada. At the same time, its economy has kept pace with ...

  10. Post-fire hydrologic response in Central Portugal. A four years study at microplot scale.

    Science.gov (United States)

    Vieira, Diana; Malvar, Maruxa; Martins, Martinho; Machado, Ana; Nunes, João; Keizer, Jacob

    2014-05-01

    Wildfires are a natural phenomenon in regions with a Mediterranean-type climate. However, their present-day widespread occurrence in southern Europe is unprecedented and strongly reflects human activity such as ignition, land-use changes, land abandonment and introduction of highly flammable plantations. Besides wildfires, post-fire management practices such as plowing, terracing, clearcutting and logging should also be considered, since their occurrence is getting increasingly common. And, in a long-term period these practices seem to be executed intercalated with repeated fire occurrences in the same site, sharing the impacts together with fire in an escalated degradational effect. In this sense, the work presented here concerns four years of runoff and erosion data at microplot scale after the wildfire, comparing different land management practices that occurred before the fire. Preliminary results indicate that in four years of monitoring, runoff is constantly higher in plowed sites than in the unplowed ones, with the exception of the first year. Regarding soil losses the plowed plots present always higher sediment rates than the unplowed ones. The comparison between two unplowed sites with different land uses, indicate higher runoff and erosion risk for pine comparatively to the eucalypt ones, however the reduced soil depth in the first can have an important role in these differences. Following these facts, the aim of the present work is to answer the following research questions: i) Do these four years of observations fit with the window of disturbance model presented by Prosser and Williams (1998). or the alternative version by Wittenberg and Inbar (2009)?; ii) Does pre-fire disturbances (wildfire, land use changes and land management practices) still have repercussions after wildfire?; In what sense does four years of intensive monitoring provides that one year couldn't provide?

  11. Do Cold Feet Warn of Trouble Ahead? Premarital Uncertainty and Four-Year Marital Outcomes

    OpenAIRE

    Lavner, Justin A.; Karney, Benjamin R.; Bradbury, Thomas N.

    2012-01-01

    Are the doubts that people feel prior to marriage signs of impending difficulties or normative experiences that can be safely ignored? To test these opposing views, we asked 464 recently-married spouses whether they had ever been uncertain about getting married and then compared four-year divorce rates and marital satisfaction trajectories among those partners with and without premarital doubts. Doubts were reported by at least one partner in two-thirds of couples. Women with premarital doubt...

  12. Contributions of Two-Year Institutions to Four-Year Completions. Snapshot™ Report, Spring 2015

    Science.gov (United States)

    National Student Clearinghouse, 2015

    2015-01-01

    In the 2013-14 academic year, 46 percent of students who completed a degree at a four-year institution were enrolled at a two-year institution at some point in the previous 10 years. This is a one percentage point increase over the comparable figure for degrees awarded in 2010-11. The prior two-year enrollment may have been brief (as little as a…

  13. Four-year dynamics of health indicators among the University students who practice Hatha Yoga regularly

    OpenAIRE

    Tolcheva, Hanna; Dudkina, Hanna; Dudkin, Mykhailo

    2016-01-01

    Purpose: to determine the dynamics of a four-year university students in health indicators, regular practice hatha-yoga. Material & Methods: the methods of theoretical analysis, synthesis and compilation of information about the study, physiological and educational testing, pedagogical experiment, methods of mathematical statistics. For experimental verification of developed on hatha-yoga program was formed by a group of 30 students enrolled in the first course of a classical university a...

  14. Enactment of third-party punishment by four-year-olds

    Directory of Open Access Journals (Sweden)

    Ben eKenward

    2012-10-01

    Full Text Available When prompted, preschoolers advocate punishment for moral transgressions against third parties, but little is known about whether and how they might act out such punishment. In this study, adult demonstrators enacted doll stories in which a perpetrator child doll made an unprovoked attack on a victim child doll, after which an adult doll punished either the perpetrator (consistent punishment or victim (inconsistent punishment. When asked to help retell the story, given free choice of their own preferred actions for the adult doll, four-year-olds (N = 32 were influenced by the demonstrated choice of target when selecting a target for punishment or admonishment. This influence was weak following inconsistent punishment, however, because the participants tended to change the story by punishing or admonishing the perpetrator when the demonstrator had punished the victim. Four-year-olds’ tendency to select a moral rule violator as a target for punishment is therefore stronger than their tendency to copy the specific actions of adults, which itself is known to be very strong. The evidence suggests that four-year-olds’ enactment of punishment is at least partially based on a belief that antisocial actions deserve to be punished.

  15. The four-year dynamics health indicators of students universities regularly practicing hatha-yoga

    Directory of Open Access Journals (Sweden)

    Hanna Tolchieva

    2016-04-01

    Full Text Available Purpose: to determine the dynamics of a four-year university students in health indicators, regular practice hatha-yoga. Material & Methods: the methods of theoretical analysis, synthesis and compilation of information about the study, physiological and educational testing, pedagogical experiment, methods of mathematical statistics. For experimental verification of developed on hatha-yoga program was formed by a group of 30 students enrolled in the first course of a classical university and expressed a desire for a four-year training visit this section three times a week. Age of students before the start of the experiment was 17–18 years old and they did not have special physical training. Results: it was found informative characteristics, which is possible to appreciate the level of health in the student's age. The content of training programs on hatha-yoga for university students who do not have special physical training. Experimentally tested program developed by hatha-yoga with students in universities for four years and recorded the dynamics of their health indicators. Conclusions: quantitative data of the study show the positive impact of regular exercise on hatha-yoga on the body of students. Positive dynamics is set according to the normalization of blood pressure and heart rate, improving posture, reducing excess body weight, improving physical qualities, enhancing the characteristics of the respiratory system.

  16. Effects of extracellular matrix proteins in chondrocyte-derived matrices on chondrocyte functions.

    Science.gov (United States)

    Hoshiba, Takashi; Lu, Hongxu; Kawazoe, Naoki; Yamada, Tomoe; Chen, Guoping

    2013-01-01

    Loss of cartilaginous phenotype during in vitro expansion culture of chondrocytes is a major barrier to the application of chondrocytes for tissue engineering. In previous study, we showed that dedifferentiation of chondrocytes during the passage culture was delayed by matrices formed by primary chondrocytes (P0-ECM). In this study, we investigated bovine chondrocyte functions when being cultured on isolated extracellular matrix (ECM) protein-coated substrata and P0-ECM. Low chondrocyte attachment was observed on aggrecan-coated substratum and P0-ECM. Cell proliferation on aggrecan- and type II collagen/aggrecan-coated substrata and P0-ECM was lower than that on the other ECM protein (type I collagen and type II collagen)-coated substrata. When chondrocytes were subcultured on aggrecan-coated substratum, decline of cartilaginous gene expression was delayed, which was similar to the cells subcultured on P0-ECM. These results indicate that aggrecan plays an important role in the regulation of chondrocyte functions and P0-ECM may be a good experimental control for investigating the role of each ECM protein in cartilage ECM.

  17. MR appearance of autologous chondrocyte implantation in the knee: correlation with the knee features and clinical outcome

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Tomoki [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Kumamoto University, Department of Orthopaedic and Neuro-Musculoskeletal Surgery, Kumamoto (Japan); Tins, Bernhard; McCall, Iain W.; Ashton, Karen [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Robert Jones and Agnes Hunt Orthopaedic Hospital NHS Trust, Department of Diagnostic Imaging, Oswestry, Shropshire (United Kingdom); Richardson, James B. [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); RJAH Orthopaedic Hospital, Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Takagi, Katsumasa [Department of Radiology and Institute of Orthopaedics, Oswestry, Shropshire (United Kingdom); Kumamoto Aging Research Institute, Kumamoto (Japan)

    2006-01-01

    To relate the magnetic resonance imaging (MRI) appearance of autologous chondrocyte implantation (ACI) in the knee in the 1st postoperative year with other knee features on MRI and with clinical outcome. Forty-nine examinations were performed in 49 patients at 1 year after ACI in the knee. Forty-one preoperative magnetic resonance (MR) examinations were also available. The grafts were assessed for smoothness, thickness in comparison with that of adjacent cartilage, signal intensity, integration to underlying bone and adjacent cartilage, and congruity of subchondral bone. Presence of overgrowth and bone marrow appearance beneath the graft were also assessed. Presence of osteophyte formation, further cartilage defects, appearance of the cruciate ligaments and the menisci were also recorded. An overall graft score was constructed, using the graft appearances. This was correlated with the knee features and the Lysholm score, a clinical self-assessment score. The data were analysed by a Kruskal-Wallis H test followed by a Mann-Whitney U test with Bonferroni correction as post-hoc test. Of 49 grafts, 32 (65%) demonstrated complete defect filling 1 year postoperatively. General overgrowth was seen in eight grafts (16%), and partial overgrowth in 13 grafts (26%). Bone marrow change underneath the graft was seen; oedema was seen in 23 grafts (47%), cysts in six grafts (12%) and sclerosis in two grafts (4%). Mean graft score was 8.7 (of maximal 12) (95% CI 8.0-9.5). Knees without osteophyte formation or additional other cartilage defects (other than the graft site) had a significantly higher graft score than knees with multiple osteophytes (P=0.0057) or multiple further cartilage defects (P=0.014). At 1 year follow-up improvement in the clinical scores was not significantly different for any subgroup. (orig.)

  18. Report on the green paper on energy. Four years of European initiatives

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2005-07-01

    In December 2000, the European Commission adopted a Green Paper on European Union energy policy. This brochure takes stock of the action undertaken in the following areas over the last four years:managing demand, diversifying internal energy sources,developing the internal energy market and the security of external supply. It presents: the Green Paper stakes, the progress made, the four political challenges, managing demand, diversifying European sources, the streamlined internal energy market, controlling external supply, future prospects for the Union, legislative developments and ten possibilities for economical energy use. (A.L.B.)

  19. Introduction of structural aseismic research in China in recent four years

    Institute of Scientific and Technical Information of China (English)

    罗奇峰; 石春香; 曹炳政; 杨树龙; 熊华; 李仕栋

    2003-01-01

    The paper mainly summarized the developments on structural aseismic theory, aseismic analysis and design ofreinforced concrete structure, lifeline system, several another kinds of structures, site and structure foundation,structure mitigation and isolation of vibration in China in recent four years. This is the introduction of recent re-search results of Chinese professionals for international organizations and professionals. At the same time, itprovides numerous abstract materials for colleagues to realize the trend of the structural aseismic theory and re-search range needing more study.

  20. Epilation for minor trachomatous trichiasis: four-year results of a randomised controlled trial.

    Directory of Open Access Journals (Sweden)

    Esmael Habtamu

    2015-03-01

    Full Text Available Trachomatous trichiasis (TT needs to be managed to reduce the risk of vision loss. The long-term impact of epilation (a common traditional practice of repeated plucking of lashes touching the eye in preventing visual impairment and corneal opacity from TT is unknown. We conducted a randomized controlled trial of epilation versus surgery for the management of minor TT (fewer than six lashes touching the eye in Ethiopia. Here we report the four-year outcome and the effect on vision and corneal opacity.1300 individuals with minor TT were recruited and randomly assigned to quality trichiasis surgery or repeated epilation using high quality epilation forceps by a trained person with good near vision. Participants were examined six-monthly for two-years, and then at four-years after randomisation. At two-years all epilation arm participants were offered free surgery. At four-years 1151 (88.5% were re-examined: 572 (88% and 579 (89% from epilation and surgery arms, respectively. At that time, 21.1% of the surgery arm participants had recurrent TT; 189/572 (33% of the epilation arm had received surgery, while 383 (67% declined surgery and had continued epilating ("epilation-only". Among the epilation-only group, 207 (54.1% fully controlled their TT, 166 (43.3% had minor TT and 10 (2.6% had major TT (>5 lashes. There were no differences between participants in the epilation-only, epilation-to-surgery and surgery arm participants in changes in visual acuity and corneal opacity between baseline and four-years.Most minor TT participants randomised to the epilation arm continued epilating and controlled their TT. Change in vision and corneal opacity was comparable between surgery and epilation-only participants. This suggests that good quality epilation with regular follow-up is a reasonable second-line alternative to surgery for minor TT for individuals who either decline surgery or do not have immediate access to surgical treatment.

  1. A four-year surveillance program for detection of Plasmodium falciparum chloroquine resistance in Honduras

    Directory of Open Access Journals (Sweden)

    Gustavo A Fontecha

    2014-07-01

    Full Text Available Countries could use the monitoring of drug resistance in malaria parasites as an effective early warning system to develop the timely response mechanisms that are required to avert the further spread of malaria. Drug resistance surveillance is essential in areas where no drug resistance has been reported, especially if neighbouring countries have previously reported resistance. Here, we present the results of a four-year surveillance program based on the sequencing of the pfcrt gene of Plasmodium falciparum populations from endemic areas of Honduras. All isolates were susceptible to chloroquine, as revealed by the pfcrt “CVMNK” genotype in codons 72-76.

  2. A four-year surveillance program for detection of Plasmodium falciparum chloroquine resistance in Honduras.

    Science.gov (United States)

    Fontecha, Gustavo A; Sanchez, Ana L; Mendoza, Meisy; Banegas, Engels; Mejía-Torres, Rosa E

    2014-07-01

    Countries could use the monitoring of drug resistance in malaria parasites as an effective early warning system to develop the timely response mechanisms that are required to avert the further spread of malaria. Drug resistance surveillance is essential in areas where no drug resistance has been reported, especially if neighbouring countries have previously reported resistance. Here, we present the results of a four-year surveillance program based on the sequencing of the pfcrt gene of Plasmodium falciparum populations from endemic areas of Honduras. All isolates were susceptible to chloroquine, as revealed by the pfcrt "CVMNK" genotype in codons 72-76.

  3. Four Years of Real-Time GRB Followup by BOOTES-1B (2005–2008

    Directory of Open Access Journals (Sweden)

    Martin Jelínek

    2010-01-01

    Full Text Available Four years of BOOTES-1B GRB follow-up history are summarised for the first time in the form of a table. The successfully followed events are described case by case. Further, the data are used to show the GRB trigger rate in Spain on a per-year basis, resulting in an estimate of 18 triggers and about 51 hours of telescope time per year for real-time triggers. These numbers grow to about 22 triggers and 77 hours per year if we include also the GRBs observable within 2 hours after the trigger.

  4. Pyridoxine neuropathy. A four-year electrophysiological and clinical follow-up of a severe case.

    Science.gov (United States)

    Santoro, L; Ragno, M; Nucciotti, R; Barbieri, F; Caruso, G

    1991-02-01

    In a 54-year-old male a severe sensory neuropathy was observed during treatment for pulmonary tuberculosis with isoniazid (400 mg/day) and pyridoxine (600 mg/day). Eight months after withdrawal from isoniazid the sensory symptomatology was still progressing, although muscle strength was never reduced. A sural nerve biopsy revealed marked loss of large myelinated fibres. Only when pyridoxine treatment was interrupted did a slow improvement begin. A clinical and electrophysiologic follow-up showed a very slow and still incomplete recovery after four years. The possibility of an unusual individual susceptibility to toxic effects of pyridoxine is considered.

  5. A FOUR-YEAR CHRONOLOGY WITH NATIONAL TEAM BOXING IN CANADA

    Directory of Open Access Journals (Sweden)

    Robert J. Schinke

    2007-10-01

    Full Text Available Applied sport psychologists tend to begin their consulting relationships with national teams having formalized skills, and often, limited contextual and sport- specific understanding. The present report overviews the first four years of a long-term consulting relationship one practitioner developed with the Canadian National Boxing Team. From the vantage of an applied sport psychology consultant, I overview how a limited consulting role expanded into increased responsibilities and opportunities. Suggestions are provided for the aspiring sport psychology consultant interested in working with athletes and coaches within the combative sport of elite amateur boxing

  6. Cartilage tissue engineering using pre-aggregated human articular chondrocytes

    Directory of Open Access Journals (Sweden)

    F Wolf

    2008-12-01

    Full Text Available In this study, we first aimed at determining whether human articular chondrocytes (HAC proliferate in aggregates in the presence of strong chondrocyte mitogens. We then investigated if the aggregated cells have an enhanced chondrogenic capacity as compared to cells cultured in monolayer. HAC from four donors were cultured in tissue culture dishes either untreated or coated with 1% agarose in the presence of TGFb-1, FGF-2 and PDGF-BB. Proliferation and stage of differentiation were assessed by measuring respectively DNA contents and type II collagen mRNA. Expanded cells were induced to differentiate in pellets or in Hyaff®-11 meshes and the formed tissues were analysed biochemically for glycosaminoglycans (GAG and DNA, and histologically by Safranin O staining. The amount of DNA in aggregate cultures increased significantly from day 2 to day 6 (by 3.2-fold, but did not further increase with additional culture time. Expression of type II collagen mRNA was about two orders of magnitude higher in aggregated HAC as compared to monolayer expanded cells. Pellets generated by aggregated HAC were generally more intensely stained for GAG than those generated by monolayer-expanded cells. Scaffolds seeded with aggregates accumulated more GAG (1.3-fold than scaffolds seeded with monolayer expanded HAC. In conclusion, this study showed that HAC culture in aggregates does not support a relevant degree of expansion. However, aggregation of expanded HAC prior to loading into a porous scaffold enhances the quality of the resulting tissues and could thus be introduced as an intermediate culture phase in the manufacture of engineered cartilage grafts.

  7. Increasing resistance to quinolones: A four-year prospective study of urinary tract infection pathogens

    Directory of Open Access Journals (Sweden)

    Orhiosefe Omigie

    2009-08-01

    Full Text Available Orhiosefe Omigie, Lawrence Okoror, Patience Umolu, Gladys IkuuhDepartment of Microbiology, Ambrose Alli University, Ekpoma, NigeriaAbstract: A four-year prospective study was carried out to determine the incidence and rate of development of resistance by common urinary tract infection (UTI pathogens to quinolone antimicrobial agents. Results show that there is high intrinsic resistance to the quinolones among strains of Pseudomonas aeruginosa (43.4%, Escherichia coli (26.3%, and Proteus spp. (17.1%. Over four years, rising rates of resistance were observed in P. aeruginosa (14.6% increase, Staphylococcus aureus (9.8%, and E. coli (9.7%. The highest potency was exhibited by ciprofloxacin (91.2%, levofloxacin (89.2%, and moxifloxacin (85.1%, while there were high rates of resistance to nalidixic acid (51.7% and pefloxacin (29.0%. Coliforms, particularly E. coli (>45%, remain the most prevalent causative agents of UTI while females within the age range of 20–50 years were most vulnerable to UTI.Keywords: UTI, microorganisms, antibiotics, resistance

  8. Local Interstellar Hydrogen's Disappearance at 1 Au: Four Years of IBEX in the Rising Solar Cycle

    CERN Document Server

    Saul, Lukas; Fuselier, Stephen; Kubiak, Marzena; McComas, Dave; Möbius, Eberhard; Sokół, Justina; Rodríguez, Diego; Scheer, Juergen; Wurz, Peter

    2013-01-01

    NASA's Interstellar Boundary Explorer (IBEX) mission has recently opened a new window on the interstellar medium (ISM) by imaging neutral atoms. One "bright" feature in the sky is the interstellar wind flowing into the solar system. Composed of remnants of stellar explosions as well as primordial gas and plasma, the ISM is by no means uniform. The interaction of the local ISM with the solar wind shapes our heliospheric environment with hydrogen being the dominant component of the very local ISM. In this paper, we report on direct sampling of the neutral hydrogen of the local ISM over four years of IBEX observations. The hydrogen wind observed at 1 AU has decreased and nearly disappeared as the solar activity has increased over the last four years; the signal at 1 AU has dropped off in 2012 by a factor of ~8 to near background levels. The longitudinal offset has also increased with time presumably due to greater radiation pressure deflecting the interstellar wind. We present longitudinal and latitudinal arriva...

  9. Follow-up of preterm children: II. Growth and development at four years of age.

    Science.gov (United States)

    Forslund, M; Bjerre, I

    1990-11-01

    In a long-term prospective study, 46 unselected infants born before 35 completed weeks of gestation were compared with 26 full-term infants. At four years of age, 44 preterms and 25 full-terms were available to follow-up. The preterms were somewhat shorter in stature and lighter in weight than the full-terms but fell well within the normal range for Swedish children. Head circumference in the preterm group was significantly correlated to neurological development. Psychometric evaluation with Griffiths' mental developmental scale showed the preterms to fall within the normal range, though their performance was inferior to that of the full-terms. The greatest differences between the groups were in scores for the subscales eye and hand coordination, performance, and practical reasoning. There was no correlation within the preterm group between the test results and birthweight, gestational age, prenatal score, perinatal score, or parental social status or education. Language development was delayed among the preterms, more of whom required speech therapy. Visual and hearing disorders were more common among preterms than expected. Although they were all in good health, more preterms than full-terms needed hospital care during the first four years of life.

  10. Overweight among Four-Year-Old Children in Relation to Early Growth Characteristics and Socioeconomic Factors

    Directory of Open Access Journals (Sweden)

    Jörgen Thorn

    2010-01-01

    Full Text Available Objectives. To assess early growth characteristics and socioeconomic factors of children in relation to body mass index (BMI and presence of overweight among four-year-old children. Methods. Two Child Health Centres (CHC participated in the study. They were selected to obtain two populations of children featuring divergent socio-economic characteristics. Growth data registered at the CHCs from birth to the 4-year check-up were recovered. Overweight was defined by the BMI cut-offs established by IOTF. BMI values expressed as BMI standard deviation score (BMISDS were used for analysis. Results. At the 4-year check-up, the BMISDS and the proportion of children with overweight (including the obese were significantly higher in the district with lower socio-economic status. High BMI at birth and low socio-economic status of the population in the CHC-district were shown to be independent determinants for overweight and BMISDS at four years of age. Conclusions. More research is needed to understand the mechanisms and how intervention programs should be designed in order to prevent the development of overweight and obesity in children.

  11. A Comparison of Four-Year Health Outcomes following Combat Amputation and Limb Salvage

    Science.gov (United States)

    Walker, Jay; Bhatnagar, Vibha; Richard, Erin; Sechriest, V. Franklin; Galarneau, Michael

    2017-01-01

    Little research has described the long-term health outcomes of patients who had combat-related amputations or leg-threatening injuries. We conducted retrospective analysis of Department of Defense and Department of Veterans Affairs health data for lower extremity combat-injured patients with (1) unilateral amputation within 90 days postinjury (early amputation, n = 440), (2) unilateral amputation more than 90 days postinjury (late amputation, n = 78), or (3) leg-threatening injuries without amputation (limb salvage, n = 107). Patient medical records were analyzed for four years postinjury. After adjusting for group differences, early amputation was generally associated with a lower or similar prevalence for adverse physical and psychological diagnoses (e.g., pain, osteoarthritis, posttraumatic stress disorder) versus late amputation and/or limb salvage. By contrast, early amputation was associated with an increased likelihood of osteoporosis during the first year postinjury. The prevalence of posttraumatic stress disorder increased for all patient groups over four years postinjury, particularly in the second year. The different clinical outcomes among combat extremity injured patients treated with early amputation, late amputation, or limb salvage highlight their different healthcare requirements. These findings can inform and optimize the specific treatment pathways that address the physical and psychological healthcare needs of such patients over time. PMID:28122002

  12. Oxygen tension affects lubricin expression in chondrocytes.

    Science.gov (United States)

    Hatta, Taku; Kishimoto, Koshi N; Okuno, Hiroshi; Itoi, Eiji

    2014-10-01

    We assessed the effects of oxygen tension on lubricin expression in bovine chondrocytes and cartilage explants and a role for hypoxia-inducible transcription factor (HIF)-1α in regulating lubricin expression was investigated using a murine chondroprogenitor cell line, ATDC5, and bovine chondrocytes isolated from superficial and middle/deep zones of femoral cartilage. ATDC5 cells and bovine chondrocytes were cultured in micromass under different oxygen tensions (21%, 5%, and 1%). ATDC5 cells and middle/deep zone chondrocytes that initially had low lubricin expression levels were also cultured with or without transforming growth factor (TGF)-β1. Quantitative reverse transcription (RT)-PCR was used to determine lubricin and chondrogenic marker gene mRNA levels and immunohistochemistry was used to assess lubricin protein expression. Explant cartilage plugs cultured under different oxygen tensions were also subjected to immunohistological analysis for lubricin. HIF-1α gene silencing was achieved by electroporatic transfer into ATDC5 cells. A low oxygen tension reduced lubricin gene expression levels in bovine superficial chondrocytes, TGF-β1-treated middle/deep zone chondrocytes, and TGF-β1-treated ATDC5 cells. Lubricin expression in explant cartilage was also suppressed under hypoxia. HIF-1α gene silencing in ATDC5 cells attenuated the lubricin expression response to the oxygen tension. These results corroborate with previous studies that the oxygen tension regulates lubricin gene expression and suggest that HIF-1α plays an important role in this regulation. The normal distribution of lubricin in articular cartilage may be due to the hypoxic oxygen environment of cartilage as it is an avascular tissue. An oxygen tension gradient may be a key factor for engineering cartilage tissue with a layered morphology.

  13. Variation across Hispanic Immigrant Generations in Parent Social Capital, College-Aligned Actions, and Four-Year College Enrollment

    Science.gov (United States)

    Ryan, Sarah; Ream, Robert K.

    2016-01-01

    Not beginning college at a four-year institution has been demonstrated as one key obstacle to equitable rates of bachelor's degree attainment among Hispanic individuals in the United States. Drawing on nationally representative longitudinal data and social capital theory, this research investigates the process of four-year college enrollment among…

  14. Dietary Fat Intake and Exercise among Two- and Four-Year College Students: Differences in Behavior and Psychosocial Factors

    Science.gov (United States)

    Berg, Carla J.; An, Lawrence C.; Ahluwalia, Jasjit S.

    2013-01-01

    Given the demographic differences among two-year colleges and four-year universities and the relatively limited access to health education and campus-based health resources, this study compares the frequency of limiting dietary fat intake and exercising among two- and four-year college students. A total of 2,265 undergraduate students aged 18-25…

  15. The Educational Attainment of Chicago Public Schools Students: A Focus on Four-Year College Degrees. Research Brief

    Science.gov (United States)

    Healey, Kaleen; Nagaoka, Jenny; Michelman, Valerie

    2014-01-01

    A four-year college degree increases a student's employment prospects and earning potential. Given this importance, it is not surprising that 75 percent of Chicago Public Schools (CPS) high school students aspire to obtain at least a four-year college degree. Yet, prior research showed that few achieved this goal: a 2006 University of Chicago…

  16. Liver Metastasis Four Years after Whipple's Resection for Solid-Pseudopapillary Tumor of the Pancreas

    Directory of Open Access Journals (Sweden)

    Srikrishna Nagri

    2007-03-01

    Full Text Available Context Solid-pseudopapillary tumor of the pancreas is a rare tumor which usually affects young females in their second and third decade of life. Metastasis is very rare after a resection of curative intent. Case report We report a case of a 65-yearold white female who presented with metastasis to the liver four years after Whipple’s resection for a solid-pseudopapillary tumor of the pancreas. Conclusions Solid-pseudopapillary tumors of the pancreas can present with metastasis a long time after resection of the primary tumor. Long term close follow up of these patients should be done. The survival rate even after liver metastasis is good.

  17. The Nuclear Organization and Management Analysis Concept methodology: Four years later

    Energy Technology Data Exchange (ETDEWEB)

    Haber, S.B.; Shurberg, D.A.; Barriere, M.T.; Hall, R.E.

    1992-08-01

    The Nuclear Organization and Management Analysis Concept was first presented at the IEEE Human Factors meeting in Monterey in 1988. In the four years since that paper, the concept and its associated methodology has been demonstrated at two commercial nuclear power plants (NPP) and one fossil power plant. In addition, applications of some of the methods have been utilized in other types of organizations, and products are being developed from the insights obtained using the concept for various organization and management activities. This paper will focus on the insights and results obtained from the two demonstration studies at the commercial NPPs. The results emphasize the utility of the methodology and the comparability of the results from the two organizations.

  18. The Nuclear Organization and Management Analysis Concept methodology: Four years later

    Energy Technology Data Exchange (ETDEWEB)

    Haber, S.B.; Shurberg, D.A.; Barriere, M.T.; Hall, R.E.

    1992-01-01

    The Nuclear Organization and Management Analysis Concept was first presented at the IEEE Human Factors meeting in Monterey in 1988. In the four years since that paper, the concept and its associated methodology has been demonstrated at two commercial nuclear power plants (NPP) and one fossil power plant. In addition, applications of some of the methods have been utilized in other types of organizations, and products are being developed from the insights obtained using the concept for various organization and management activities. This paper will focus on the insights and results obtained from the two demonstration studies at the commercial NPPs. The results emphasize the utility of the methodology and the comparability of the results from the two organizations.

  19. Photon Detection Efficiency Measurements of the VERITAS Cherenkov Telescope Photomultipliers after four Years of Operation

    CERN Document Server

    Gazda, Eliza; Otte, Nepomuk; Richards, Gregory

    2016-01-01

    The photon detection efficiency of two sets of R10560-100-20 superbialkali photomultiplier tubes from Hamamatsu were measured between 200 nm and 750 nm to quantify a possible degradation of the photocathode sensitivity after four years of operation in the cameras of the VERITAS Cherenkov telescopes. A sample of 20 photomultiplier tubes, which was removed from the telescopes was compared with a sample of 20 spare photomultiplier tubes, which had been kept in storage. It is found that the average photocathode sensitivity marginally increased below 300 nm and dropped by 10% to 30% above 500 nm. The average photocathode sensitivity folded with the Cherenkov spectrum emitted by particles in air showers, however, reveals a consistent detection yield of 18.9+/-0.2% and 19.1+/-0.2% for the sample removed from the telescope and the spare sample, respectively.

  20. ACGT: advancing clinico-genomic trials on cancer - four years of experience.

    Science.gov (United States)

    Martin, Luis; Anguita, Alberto; Graf, Norbert; Tsiknakis, Manolis; Brochhausen, Mathias; Rüping, Stefan; Bucur, Anca; Sfakianakis, Stelios; Sengstag, Thierry; Buffa, Francesca; Stenzhorn, Holger

    2011-01-01

    The challenges regarding seamless integration of distributed, heterogeneous and multilevel data arising in the context of contemporary, post-genomic clinical trials cannot be effectively addressed with current methodologies. An urgent need exists to access data in a uniform manner, to share information among different clinical and research centers, and to store data in secure repositories assuring the privacy of patients. Advancing Clinico-Genomic Trials (ACGT) was a European Commission funded Integrated Project that aimed at providing tools and methods to enhance the efficiency of clinical trials in the -omics era. The project, now completed after four years of work, involved the development of both a set of methodological approaches as well as tools and services and its testing in the context of real-world clinico-genomic scenarios. This paper describes the main experiences using the ACGT platform and its tools within one such scenario and highlights the very promising results obtained.

  1. Four-Year-Olds Use a Mixture of Spatial Reference Frames.

    Directory of Open Access Journals (Sweden)

    James Negen

    Full Text Available Keeping track of unseen objects is an important spatial skill. In order to do this, people must situate the object in terms of different frames of reference, including body position (egocentric frame of reference, landmarks in the surrounding environment (extrinsic frame reference, or other attached features (intrinsic frame of reference. Nardini et al. hid a toy in one of 12 cups in front of children, turned the array when they were not looking, and then asked them to point to the cup with the toy. This forced children to use the intrinsic frame (information about the array of cups to locate the hidden toy. Three-year-olds made systematic errors by using the wrong frame of reference, 4-year-olds were at chance, and only 5- and 6-year-olds were successful. Can we better understand the developmental change that takes place at four years? This paper uses a modelling approach to re-examine the data and distinguish three possible strategies that could lead to the previous results at four years: (1 Children were choosing cups randomly, (2 Children were pointing between the egocentric/extrinsic-cued location and the correct target, and (3 Children were pointing near the egocentric/extrinsic-cued location on some trials and near the target on the rest. Results heavily favor the last possibility: 4-year-olds were not just guessing or trying to combine the available frames of reference. They were using the intrinsic frame on some trials, but not doing so consistently. These insights suggest that accounts of improving spatial performance at 4 years need to explain why there is a mixture of responses. Further application of the selected model also suggests that children become both more reliant on the correct frame and more accurate with any chosen frame as they mature.

  2. Four-Year-Olds Use a Mixture of Spatial Reference Frames.

    Science.gov (United States)

    Negen, James; Nardini, Marko

    2015-01-01

    Keeping track of unseen objects is an important spatial skill. In order to do this, people must situate the object in terms of different frames of reference, including body position (egocentric frame of reference), landmarks in the surrounding environment (extrinsic frame reference), or other attached features (intrinsic frame of reference). Nardini et al. hid a toy in one of 12 cups in front of children, turned the array when they were not looking, and then asked them to point to the cup with the toy. This forced children to use the intrinsic frame (information about the array of cups) to locate the hidden toy. Three-year-olds made systematic errors by using the wrong frame of reference, 4-year-olds were at chance, and only 5- and 6-year-olds were successful. Can we better understand the developmental change that takes place at four years? This paper uses a modelling approach to re-examine the data and distinguish three possible strategies that could lead to the previous results at four years: (1) Children were choosing cups randomly, (2) Children were pointing between the egocentric/extrinsic-cued location and the correct target, and (3) Children were pointing near the egocentric/extrinsic-cued location on some trials and near the target on the rest. Results heavily favor the last possibility: 4-year-olds were not just guessing or trying to combine the available frames of reference. They were using the intrinsic frame on some trials, but not doing so consistently. These insights suggest that accounts of improving spatial performance at 4 years need to explain why there is a mixture of responses. Further application of the selected model also suggests that children become both more reliant on the correct frame and more accurate with any chosen frame as they mature.

  3. Atypical hemolytic uremic syndrome diagnosed four years after ABO-incompatible kidney transplantation.

    Science.gov (United States)

    Kawaguchi, Keiko; Kawanishi, Kunio; Sato, Masayo; Itabashi, Mitsuyo; Fujii, Akiko; Kanetsuna, Yukiko; Huchinoue, Shouhei; Ohashi, Ryuji; Koike, Junki; Honda, Kazuho; Nagashima, Yoji; Nitta, Kosaku

    2015-07-01

    Atypical hemolytic uremic syndrome (aHUS) in allograft kidney transplantation is caused by various factors including rejection, infection, and immunosuppressive drugs. We present a case of a 32 year old woman with aHUS four years after an ABO-incompatible kidney transplantation from a living relative. The primary cause of end-stage renal disease was unknown; however, IgA nephropathy (IgAN) was suspected from her clinical course. She underwent pre-emptive kidney transplantation from her 60 year old mother. The allograft preserved good renal function [serum creatinine (sCr) level 110-130 μmol/L] until a sudden attack of abdominal pain four years after transplant, with acute renal failure (sCr level, 385.3 μmol/L), decreasing platelet count, and hemolytic anemia with schizocytes. On allograft biopsy, there was thrombotic microangiopathy in the glomeruli, with a cellular crescent formation and mesangial IgA and C3 deposition. Microvascular inflammation, such as glomerulitis, peritubular capillaritis, and arteriole endarteritis were also detected. A disintegrin-like and metalloproteinase with thrombospondin type 1 motifs 13 (ADAMTS13) did not decrease and Shiga toxin was not detected. Donor-specific antibodies or autoantibodies, including anti-neutrophil cytoplasmic antibody and anti-glomerular basement membrane (anti-GBM) antibody, were negative. The patient was diagnosed with aHUS and received three sessions of plasmapheresis and methylprednisolone pulse therapy, followed by oral methylprednisolone (0.25-0.5 mg/kg) instead of tacrolimus. She temporarily required hemodialysis (sCr level, 658.3 μmol/L). Thereafter, her sCr level improved to 284.5 μmol/L without dialysis therapy. This case is clinically considered as aHUS after kidney transplantation, associated with various factors, including rejection, glomerulonephritis, and toxicity from drugs such as tacrolimus.

  4. Increased chondrocyte adhesion on nanotubular anodized titanium.

    Science.gov (United States)

    Burns, Kevin; Yao, Chang; Webster, Thomas J

    2009-03-01

    Previous studies have demonstrated increased osteoblast (bone-forming cells) functions (including adhesion, synthesis of intracellular collagen, alkaline phosphatase activity, and deposition of calcium-containing minerals) on titanium anodized to possess nanometer features compared with their unanodized counterparts. Such titanium materials were anodized to possess novel nanotubes also capable of drug delivery. Since titanium has not only experienced wide spread commercial use in orthopedic but also in cartilage applications, the objective of the present in vitro study was for the first time to investigate chondrocyte (cartilage synthesizing cells) functions on titanium anodized to possess nanotubes. For this purpose, titanium was anodized in dilute hydrofluoric acid at 20 V for 20 min. Results showed increased chondrocyte adhesion on anodized titanium with nanotube structures compared with unanodized titanium. Importantly, the present study also provided evidence why. Since material characterization studies revealed significantly greater nanometer roughness and similar chemistry as well as crystallinity between nanotubular anodized and unanodized titanium, the results of the present study highlight the importance of the nanometer roughness provided by anodized nanotubes on titanium for enhancing chondrocyte adhesion. In this manner, the results of the present in vitro study indicated that anodization might be a promising quick and inexpensive method to modify the surface of titanium-based implants to induce better chondrocyte adhesion for cartilage applications.

  5. Delineation and Scale Effect of Precision Agriculture Management Zones Using Yield Monitor Data Over Four Years

    Institute of Scientific and Technical Information of China (English)

    LI Xiang; PAN Yu-chun; GE Zhong-qiang; ZHAO Chun-jiang

    2007-01-01

    In this study, precision agriculture management zones were delineated using yield data over four years from the combine harvester equipped with yield monitor and DGPS receiver. Relative yields measured during each year were interpolated to 4 m2 grid size using ordinary kriging. The resultant interpolated yield maps were averaged across years to create a map of the mean relative yield, which was then used for cluster analysis. The mean yield map of post-classification was processed by applying majority filtering with window sizes that were equivalent to the grid sizes of 12, 20, 28, 36, 44, 52 and 60 m. The scale effect of management zones was evaluated using relative variance reduction, test of significant differences of the means of yield zones, spatial fragmentation, and spatial agreement. The results showed that the post-classification majority filtering (PCMF) eliminated lots of isolated cells or patches caused by random variation while preserving yield means, high variance reduction, general yield patterns, and high spatial agreement. The zoned result can be used as yield goal map for preplant or in-season fertilizer recommendation in precision agriculture.

  6. FOUR-YEAR-OLD NAMSAN TUNNEL CONGESTION PRICING SCHEME IN SEOUL

    Directory of Open Access Journals (Sweden)

    Bongsoo SON, Ph. D.

    2002-01-01

    Full Text Available The purpose of this paper is to evaluate the effectiveness of the congestion pricing scheme at Namsan #1 and #3 tunnels in downtown Seoul four years after its implementation. The effectiveness of the scheme was measured by the changes of various traffic impacts. The traffic volume of the two tunnels was reduced by up to 25% for the first month. After that time, the traffic volume started to increase again and then exceeded the previous volume level. However, average travel speed of the two tunnel corridors improved by up to 74%. The overall traffic volume of the four alternative routes was increased; nevertheless, their average travel speed increased as well. The number of carpool vehicles occupied by 3 or more persons including the driver during the peak periods was remarkably increased. Before the congestion fee charging, toll-charged vehicles amounted to 68.5% of the total traffic volume of the two tunnels, and then the share dropped to 29% afterwards. The empirical analysis results for the effectiveness of the congestion pricing scheme are very promising.

  7. The Impact of Impulsivity on Weight Loss Four Years after Bariatric Surgery

    Directory of Open Access Journals (Sweden)

    Kathrin Schag

    2016-11-01

    Full Text Available Bariatric surgery has serious implications on metabolic health. The reasons for a failure of bariatric surgery, i.e., limited weight loss, are multifactorial and include psychological factors. We established a theoretical model of how impulsivity is related to weight loss outcome. We propose that depressive symptoms act as a mediator between impulsivity and pathological eating behavior, and that pathological eating behavior has a direct impact on weight loss outcome. We calculated excessive weight loss (%EWL and assessed self-reported impulsivity (using the Baratt Impulsiveness Scale (BIS-15 total score, depressive symptoms (the Patient Health Questionnaire (PHQ-9 score, and pathological eating behavior (the Eating Disorder Inventory 2 (EDI-2 total score in 65 patients four years after laparoscopic sleeve gastrectomy. Regression and mediation analyses were computed to validate the theoretical model. The BIS-15, PHQ-9, and EDI-2 have medium to high correlations between each other, and EDI-2 correlated with %EWL. The mediation analysis yielded that the PHQ-9 represents a significant mediator between BIS-15 and EDI-2. The regression model between EDI-2 and %EWL was also significant. These results support our theoretical model, i.e., suggest that impulsivity has an indirect impact on weight loss outcome after bariatric surgery, mediated by depression and transferred through pathological eating behavior. Thus, the underlying psychological factors should be addressed in post-operative care to optimize weight loss outcome.

  8. The Effects of Inquiry-Based Integrated Information Literacy Instruction: Four-Year Trends

    Directory of Open Access Journals (Sweden)

    Lin Ching Chen

    2014-07-01

    Full Text Available The purpose of this study was to examine the effects of four-year integrated information literacy instruction via a framework of inquiry-based learning on elementary students’ memory and comprehension. Moderating factors of students’ academic achievement was another focus of this study. The subjects were 72 students who have participated in this study since they entered an elementary school in Chiayi district. This elementary school adopted the integrated information literacy instruction, designed by the researchers and elementary school teachers, and integrated it into various subject matters via a framework of inquiry-based learning, such as Super 3 and Big6 models. A series of inquiry-based integrated information literacy instruction has been implemented since the second semester of the subjects’ first grade. A total of seven inquiry learning projects has been implemented from grade one through grade four. Fourteen instruments were used as pretests and posttests to assess students’ factual recall and conceptual understanding of subject contents in different projects. The results showed that inquiry-based integrated information literacy instruction couldhelp students memorize facts and comprehend concepts of subject contents. Regardless ofacademic achievements, if students would like to devote their efforts to inquiry processes, their memory and comprehension of subject contents improvedeffectively. However, students of low-academic achievement might need more time to be familiar with the inquiry-based learning strategy.

  9. THE FERMI GBM GAMMA-RAY BURST SPECTRAL CATALOG: FOUR YEARS OF DATA

    Energy Technology Data Exchange (ETDEWEB)

    Gruber, David; Von Ahlefeld, Victoria Weller; Diehl, Roland; Greiner, Jochen; Von Kienlin, Andreas [Max-Planck-Institut für extraterrestrische Physik, Giessenbachstrasse 1, D-85748 Garching (Germany); Goldstein, Adam; Bhat, P. Narayana; Briggs, Michael S.; Connaughton, Valerie [University of Alabama in Huntsville, 320 Sparkman Drive, Huntsville, AL 35805 (United States); Bissaldi, Elisabetta [Istituto Nazionale di Fisica Nucleare, Sezione di Trieste, I-34127 Trieste (Italy); Byrne, Dave; Fitzpatrick, Gerard; Foley, Suzanne [School of Physics, University College Dublin, Belfield, Stillorgan Road, Dublin 4 (Ireland); Cleveland, William H. [Universities Space Research Association, 320 Sparkman Drive, Huntsville, AL 35805 (United States); Fishman, Gerald J.; Kouveliotou, Chryssa [Space Science Office, VP62, NASA/Marshall Space Flight Center, Huntsville, AL 35812 (United States); Gibby, Melissa; Giles, Misty M. [Jacobs Technology, Inc., Huntsville, AL (United States); Guiriec, Sylvain [NASA Goddard Space Flight Center, Greenbelt, MD 20771 (United States); Van der Horst, Alexander J. [Astronomical Institute, University of Amsterdam, Science Park 904, 1098 XH Amsterdam (Netherlands); and others

    2014-03-01

    In this catalog we present the updated set of spectral analyses of gamma-ray bursts (GRBs) detected by the Fermi Gamma-Ray Burst Monitor during its first four years of operation. It contains two types of spectra, time-integrated spectral fits and spectral fits at the brightest time bin, from 943 triggered GRBs. Four different spectral models were fitted to the data, resulting in a compendium of more than 7500 spectra. The analysis was performed similarly but not identically to Goldstein et al. All 487 GRBs from the first two years have been re-fitted using the same methodology as that of the 456 GRBs in years three and four. We describe, in detail, our procedure and criteria for the analysis and present the results in the form of parameter distributions both for the observer-frame and rest-frame quantities. The data files containing the complete results are available from the High-Energy Astrophysics Science Archive Research Center.

  10. The Western Kentucky University Blazar Monitoring Program: The First Four Years

    Science.gov (United States)

    Walters, R.; Barnaby, D.; Carini, M.

    2006-07-01

    In 2000, we began a blazar monitoring program utilizing the 0.6m telescope of the Bell Observatory at Western Kentucky University. We chose a sample of objects from across the αro - αox plane in order to characterize the variability characteristics of HBLs, IBLs and LBLs. The most rapid timescale variations, known as microvariability, provide information on differences in jet characteristics and observations of microvariability at different colors provides insight into the nature of the particle acceleration mechanisms in the different subclasses of blazars during different activity levels. On longer timescales (days to weeks) multiwavelength observations (via our participation in WEBT campaigns, VERITAS campaigns, etc) can be used to confront models describing the origin of the seed photons responsible for the observed high energy emission in these objects. Through our ability to obtain well sampled light curves, we will, at the longest timescales (weeks to decades), establish the differences in the variability properties between different sub-classes. We present here the results of the first fours years of this program.

  11. THE INFORMATION-COMMUNICATION TECHNOLOGY IN THE HOME ENVIRONMENT OF FOUR-YEAR-OLD PRESCHOOL CHILDREN

    Directory of Open Access Journals (Sweden)

    Jurka Lepičnik Vodopivec

    2014-01-01

    Full Text Available Today, preschool children live in an era of information-communication technologies (ICT. There exist various definitions of ICT. For the purposes of our research we thus used a broader definition of ICT, which stretches beyond mobiles and computers, and includes a variety of everyday technologies. Children already encounter them in their family environment. We wanted to find out which ICT children use the most, how they access them and if ICT has any influence on their development. Our sample consisted of 130 parents of four-year-old children who visit kindergarten. We were interested in the adults' – parents' influence and in the influence of the children’s gender on the access to and usage of ICT. We similarly wanted to know if these are influenced by the toy industry market and family values. A part of the research is also dedicated to the ICT’s influence on the perception of childhood and a plausible occurrence of the digital divide.

  12. The Fermi GBM Gamma-Ray Burst Spectral Catalog: Four Years Of Data

    CERN Document Server

    Gruber, David; von Ahlefeld, Victoria Weller; Bhat, P Narayana; Bissaldi, Elisabetta; Briggs, Michael S; Byrne, Dave; Cleveland, William H; Connaughton, Valerie; Fishman, Gerald J; Fitzpatrick, Gerard; Foley, Suzanne; Gibby, Melissa; Giles, Misty M; Greiner, Jochen; Guiriec, Sylvain; van der Horst, Alexander J; von Kienlin, Andreas; Kouveliotou, Chryssa; Layden, Emily; Lin, Lin; Meegan, Charles A; McGlynn, Sinéad; Paciesas, William S; Pelassa, Véronique; Preece, Robert D; Rau, Arne; Wilson-Hodge, Colleen A; Xiong, Shaolin; Yu, Hoi-Fung

    2014-01-01

    In this catalog we present the updated set of spectral analyses of GRBs detected by the Fermi Gamma-Ray Burst Monitor (GBM) during its first four years of operation. It contains two types of spectra, time-integrated spectral fits and spectral fits at the brightest time bin, from 943 triggered GRBs. Four different spectral models were fitted to the data, resulting in a compendium of more than 7500 spectra. The analysis was performed similarly, but not identically to Goldstein et al. 2012. All 487 GRBs from the first two years have been re-fitted using the same methodology as that of the 456 GRBs in years three and four. We describe, in detail, our procedure and criteria for the analysis, and present the results in the form of parameter distributions both for the observer-frame and rest-frame quantities. The data files containing the complete results are available from the High-Energy Astrophysics Science Archive Research Center (HEASARC).

  13. Who Benefits From Four-Year Higher Education in Turkey? A Case Study of Law School

    Directory of Open Access Journals (Sweden)

    Filiz GÖLPEK

    2013-01-01

    Full Text Available In many countries, a higher education system which is based on welfare state practices prevails. In this system, the services are substantially provided by the state free of charge or at quite low prices. Since the services are financed by taxes, additional sources cannot be provided; and because of the fact that it doesn't entail a real cost benefit comparison by the users, it leads to inefficiency. From the early 80s on, there has been a tendency towards the marketing mechanisms in the provision of many public services such as education due to both strong theoretical and empirical reason, and the restrictions in public financing. Together with this tendency, the question of who benefits from education services is tried to be answered in order to discuss the stimulation of activities and additional resources on a reasonable base. The aim of this study is to determine whether it is the individual or the society who benefits from this public service at the higher education level by calculating private and social return rates through internal return rate technique which is one of the cost benefit analysis method. Findings from calculations have shown that the individual gains more than the society after a four-year university education.

  14. [Platelet transfusion and allergic transfusion reactions: experiences at Lille Hospital over a four year period].

    Science.gov (United States)

    Wibaut, B; Vannier, V; Renom, P; Goudemand, J

    2000-04-01

    Among the immediate transfusion reactions caused by the utilization of blood products, those suggesting immuno-allergic mechanisms posed problems for frequency, gravity, laboratory diagnosis and safety. We report here the Lille Hospital's experience over a four-year period concerning these manifestations after platelet concentrate transfusion. Eight hundred and fifty-two immediate transfusion reactions have been declared, of which 230 were allergic, which appeared in 181 patients (27%). Among the most frequent clinical signs, rash was often described (158 cases: 68.7%); less frequent were respiratory problems such as dyspnea (34 cases: 14.8%) and hypotensive reactions (18 cases: 7.8%). Seven patients presented severe reactions (3%). Twenty percent of them presented multiple allergic reactions and in 43.2%, the recurrence was more serious than the initial problem in spite of preventive medication; the use of washed blood components was necessary. The age of platelet concentrates does not appear to play a part in provoking these events (67% of platelet concentrates had been collected within four days). These allergic transfusion reactions posed problems for those who prescribe medication, because they are frequent, sometimes serious, can recur and at present, the proposed medication prevention is not always efficient.

  15. Stromal cell-derived factor 1 regulates the actin organization of chondrocytes and chondrocyte hypertrophy.

    Directory of Open Access Journals (Sweden)

    Koichi Murata

    Full Text Available Stromal cell-derived factor 1 (SDF-1/CXCL12/PBSF plays important roles in the biological and physiological functions of haematopoietic and mesenchymal stem cells. This chemokine regulates the formation of multiple organ systems during embryogenesis. However, its roles in skeletal development remain unclear. Here we investigated the roles of SDF-1 in chondrocyte differentiation. We demonstrated that SDF-1 protein was expressed at pre-hypertrophic and hypertrophic chondrocytes in the newly formed endochondral callus of rib fracture as well as in the growth plate of normal mouse tibia by immunohistochemical analysis. Using SDF-1(-/- mouse embryo, we histologically showed that the total length of the whole humeri of SDF-1(-/- mice was significantly shorter than that of wild-type mice, which was contributed mainly by shorter hypertrophic and calcified zones in SDF-1(-/- mice. Actin cytoskeleton of hypertrophic chondrocytes in SDF-1(-/- mouse humeri showed less F-actin and rounder shape than that of wild-type mice. Primary chondrocytes from SDF-1(-/- mice showed the enhanced formation of philopodia and loss of F-actin. The administration of SDF-1 to primary chondrocytes of wild-type mice and SDF-1(-/- mice promoted the formation of actin stress fibers. Organ culture of embryonic metatarsals from SDF-1(-/- mice showed the growth delay, which was recovered by an exogenous administration of SDF-1. mRNA expression of type X collagen in metatarsals and in primary chondrocytes of SDF-1(-/- mouse embryo was down-regulated while the administration of SDF-1 to metatarsals recovered. These data suggests that SDF-1 regulates the actin organization and stimulates bone growth by mediating chondrocyte hypertrophy.

  16. Bone graft

    Science.gov (United States)

    ... around the area. The bone graft can be held in place with pins, plates, or screws. Why ... Orthopaedic Surgery, San Francosco, CA. Also reviewed by David Zieve, MD, MHA, Isla Ogilvie, PhD, and the ...

  17. Stomatal response of Pinus sylvestriformis to elevated CO2 concentrations during the four years of exposure

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yu-mei; HAN Shi-jie; LIU Ying; JIA Xia

    2005-01-01

    Four-year-old Pinus sylvestriformis were exposed for four growing seasons in open top chambers to ambient CO2 concentration (approx. 350 μmol·mol-1) and high CO2 concentrations (500 and 700 μmol·mol-1) at Research Station of Changbai Mountain Forest Ecosystems, Chinese Academy of Sciences at Antu Town, Jilin Province, China (42oN, 128oE). Stomatal response to elevated CO2 concentrations was examined by stomatal conductance (gs), ratio of intercellular to ambient CO2 concentration (ci/ca) and stomatal number. Reciprocal transfer experiments of stomatal conductance showed that stomatal conductance in high-[CO2]-grown plants increased in comparison with ambient-[CO2]-grown plants when measured at their respective growth CO2 concentration and at the same measurement CO2 concentration (except a reduction in 700 μmol·mol-1 CO2 grown plants compared with plants on unchambered field when measured at growth CO2 concentration and 350 μmol·mol-1CO2). High-[CO2]-grown plants exhibited lower ci/ca ratios than ambient-[CO2]-grown plants when measured at their respective growth CO2 concentration. However, ci/ca ratios increased for plants grown in high CO2 concentrations compared with control plants when measured at the same CO2 concentration. There was no significant difference in stomatal number per unit long needle between elevated and ambient CO2. However, elevated CO2 concentrations reduced the total stomatal number of whole needle by the decline of stomatal line and changed the allocation pattern of stomata between upper and lower surface of needle.

  18. Implementation of a four-year multimedia computer curriculum in cardiology at six medical schools.

    Science.gov (United States)

    Petrusa, E R; Issenberg, S B; Mayer, J W; Felner, J M; Brown, D D; Waugh, R A; Kondos, G T; Gessner, I H; McGaghie, W C

    1999-02-01

    The pressures of a changing health care system are making inroads on the commitment and effort that both basic science and clinical faculty can give to medical education. A tool that has the potential to compensate for decreased faculty time and thereby to improve medical education is multimedia computer instruction that is applicable at all levels of medical education, developed according to instructional design principles, and supported by evidence of effectiveness. The authors describe the experiences of six medical schools in implementing a comprehensive computer-based four-year curriculum in bedside cardiology developed by a consortium of university cardiologists and educational professionals. The curriculum consisted of ten interactive, patient-centered, case-based modules focused on the history, physical examination, laboratory data, diagnosis, and treatment. While an optimal implementation plan was recommended, each institution determined its own strategy. Major goals of the project, which took place from July 1996 to June 1997, were to identify and solve problems of implementation and to assess learners' and instructors' acceptance of the system and their views of its value. A total of 1,586 students used individual modules of the curriculum 6,131 times. Over 80% of students rated all aspects of the system highly, especially its clarity and educational value compared with traditional lectures. The authors discuss the aspects of the curriculum that worked, problems that occurred (such as difficulties in scheduling use of the modules in the third year), barriers to change and ways to overcome them (such as the type of team needed to win acceptance for and oversee implementation of this type of curriculum), and the need in succeeding years to formally assess the educational effectiveness of this and similar kinds of computer-based curricula.

  19. A four-year, systems-wide intervention promoting interprofessional collaboration

    Directory of Open Access Journals (Sweden)

    Braithwaite Jeffrey

    2012-04-01

    Full Text Available Abstract Background A four-year action research study was conducted across the Australian Capital Territory health system to strengthen interprofessional collaboration (IPC though multiple intervention activities. Methods We developed 272 substantial IPC intervention activities involving 2,407 face-to-face encounters with health system personnel. Staff attitudes toward IPC were surveyed yearly using Heinemann et al's Attitudes toward Health Care Teams and Parsell and Bligh's Readiness for Interprofessional Learning scales (RIPLS. At study's end staff assessed whether project goals were achieved. Results Of the improvement projects, 76 exhibited progress, and 57 made considerable gains in IPC. Educational workshops and feedback sessions were well received and stimulated interprofessional activities. Over time staff scores on Heinemann's Quality of Interprofessional Care subscale did not change significantly and scores on the Doctor Centrality subscale increased, contrary to predictions. Scores on the RIPLS subscales of Teamwork & Collaboration and Professional Identity did not alter. On average for the assessment items 33% of staff agreed that goals had been achieved, 10% disagreed, and 57% checked neutral. There was most agreement that the study had resulted in increased sharing of knowledge between professions and improved quality of patient care, and least agreement that between-professional rivalries had lessened and communication and trust between professions improved. Conclusions Our longitudinal interventional study of IPC involving multiple activities supporting increased IPC achieved many project-specific goals. However, improvements in attitudes over time were not demonstrated and neutral assessments predominated, highlighting the difficulties faced by studies targeting change at the systems level and over extended periods.

  20. Radiation survey on Fukushima Medical University premises about four years after the Fukushima nuclear disaster

    Science.gov (United States)

    Omori, Yasutaka; Wakamatsu, Hiroaki; Sorimachi, Atsuyuki; Ishikawa, Tetsuo

    2016-01-01

    Abstract This study was conducted on the Fukushima Medical University (FMU) premises (in Fukushima City, Fukushima Prefecture) about four years after the Fukushima Daiichi Nuclear Power Plant accident. Its objectives were (1) to create a map of the ambient gamma dose rate (air-kerma rate) distribution, (2) to evaluate the air-kerma rate originating from natural radionuclides, and (3) to investigate the effects of snow cover on changes in the air-kerma rate. This man-borne survey revealed that the air-kerma rate varies widely, ranging from 0.038 μGy h-1 to 0.520 μGy h-1, and is higher on grass than on the other investigated surface types, such as soil, asphalt, and bricks. In this area, the mean air-kerma rate from natural radiation was evaluated to be 0.03 ± 0.01 μGy h-1, which is close to 0.04 μGy h-1, which was measured in central Fukushima City by a local authority.Furthermore, snowfall was found to reduce the air-kerma rate by 5%-30%. This reduction was attributed to attenuation of the primary radiation while passing through the snow cover, and the measured contribution of scattered radiation to the air-kerma rate reduction was small. The reduction rate was found to depend on the initial snow depth but to maintain a similar value for a couple of days, after the snow had partially melted and its depth had decreased. Finally, analysis of the daily dose due to external exposure received on the FMU premises revealed that no further health effects due to chronic radiation exposure at this site are to be expected. PMID:26911302

  1. Four-year record of mercury wet deposition in one typical industrial city in southwest China

    Science.gov (United States)

    Qin, Caiqing; Wang, Yongmin; Peng, Yulong; Wang, Dingyong

    2016-10-01

    During the period from December 2010 to November 2014, long-term monitoring of Hg wet deposition was conducted at three sites in Chongqing. The four-year volume-weighted mean concentrations (VWC) of total mercury (THg) and methylmercury (MeHg) in precipitation were 34.25 ng L-1 and 0.48 ng L-1, respectively. The average annual wet deposition fluxes were 37.83 ± 11.53 μg m-2 yr-1 for THg and 0.61 ± 0.19 μg m-2 yr-1 for MeHg. Besides, the average proportion of MeHg in THg was 1.41%. These data were well ahead of values observed in most of other areas. The annual VWC of Hg rose remarkably in 2012 and then reached a plateau or climbed mildly in the following 2 years, while its annual wet deposition fluxes saw an upward trend throughout the whole period. In addition, the high figures of Hg concentration were generally found in the non-monsoon seasons, but its elevated wet deposition fluxes normally appeared in the rainy seasons. These characteristics of inter-annual and seasonal changes for VWC and fluxes were observed at every sampling site in this study as well. Besides, there were significantly spatial distributions for VWC and fluxes of THg in the descending order of the downtown (NA), the suburban (BB) and the controlled site (JY). While for MeHg, BB had the largest values, followed by NA and JY. So apart from THg load, the formation of MeHg was influenced by other factors in Chongqing, like the concentration of reactive mercury (RHg) in precipitation. Additionally, particulate bound mercury (PHg) is the dominant form among various Hg species, and atmospheric Hg is effectively eliminated during the initial period of the rain event and the below-cloud scavenging is the predominant mechanism contributing Hg to precipitation.

  2. Radiation survey on Fukushima Medical University premises about four years after the Fukushima nuclear disaster.

    Science.gov (United States)

    Omori, Yasutaka; Wakamatsu, Hiroaki; Sorimachi, Atsuyuki; Ishikawa, Tetsuo

    2016-06-08

    This study was conducted on the Fukushima Medical University (FMU) premises (in Fukushima City, Fukushima Prefecture) about four years after the Fukushima Daiichi Nuclear Power Plant accident. Its objectives were (1) to create a map of the ambient gamma dose rate (air-kerma rate) distribution, (2) to evaluate the air-kerma rate originating from natural radionuclides, and (3) to investigate the effects of snow cover on changes in the air-kerma rate. This man-borne survey revealed that the air-kerma rate varies widely, ranging from 0.038 μGy h(-1) to 0.520 μGy h(-1), and is higher on grass than on the other investigated surface types, such as soil, asphalt, and bricks. In this area, the mean air-kerma rate from natural radiation was evaluated to be 0.03 ± 0.01 μGy h(-1), which is close to 0.04 μGy h(-1), which was measured in central Fukushima City by a local authority.Furthermore, snowfall was found to reduce the air-kerma rate by 5%-30%. This reduction was attributed to attenuation of the primary radiation while passing through the snow cover, and the measured contribution of scattered radiation to the air-kerma rate reduction was small. The reduction rate was found to depend on the initial snow depth but to maintain a similar value for a couple of days, after the snow had partially melted and its depth had decreased. Finally, analysis of the daily dose due to external exposure received on the FMU premises revealed that no further health effects due to chronic radiation exposure at this site are to be expected.

  3. A four-year cardiovascular risk score for type 2 diabetic inpatients

    Directory of Open Access Journals (Sweden)

    Dolores Ramírez-Prado

    2015-06-01

    Full Text Available As cardiovascular risk tables currently in use were constructed using data from the general population, the cardiovascular risk of patients admitted via the hospital emergency department may be underestimated. Accordingly, we constructed a predictive model for the appearance of cardiovascular diseases in patients with type 2 diabetes admitted via the emergency department. We undertook a four-year follow-up of a cohort of 112 adult patients with type 2 diabetes admitted via the emergency department for any cause except patients admitted with acute myocardial infarction, stroke, cancer, or a palliative status. The sample was selected randomly between 2010 and 2012. The primary outcome was time to cardiovascular disease. Other variables (at baseline were gender, age, heart failure, renal failure, depression, asthma/chronic obstructive pulmonary disease, hypertension, dyslipidaemia, insulin, smoking, admission for cardiovascular causes, pills per day, walking habit, fasting blood glucose and creatinine. A cardiovascular risk table was constructed based on the score to estimate the likelihood of cardiovascular disease. Risk groups were established and the c-statistic was calculated. Over a mean follow-up of 2.31 years, 39 patients had cardiovascular disease (34.8%, 95% CI [26.0–43.6%]. Predictive factors were gender, age, hypertension, renal failure, insulin, admission due to cardiovascular reasons and walking habit. The c-statistic was 0.734 (standard error: 0.049. After validation, this study will provide a tool for the primary health care services to enable the short-term prediction of cardiovascular disease after hospital discharge in patients with type 2 diabetes admitted via the emergency department.

  4. Chondrocyte hypertrophy in skeletal development, growth, and disease.

    Science.gov (United States)

    Sun, Margaret Man-Ger; Beier, Frank

    2014-03-01

    Most of our bones form through the process of endochondral ossification, which is tightly regulated by the activity of the cartilage growth plate. Chondrocyte maturation through the various stages of growth plate physiology ultimately results in hypertrophy. Chondrocyte hypertrophy is an essential contributor to longitudinal bone growth, but recent data suggest that these cells also play fundamental roles in signaling to other skeletal cells, thus coordinating endochondral ossification. On the other hand, ectopic hypertrophy of articular chondrocytes has been implicated in the pathogenesis of osteoarthritis. Thus, a better understanding of the processes that control chondrocyte hypertrophy in the growth plate as well as in articular cartilage is required for improved management of both skeletal growth disorders and osteoarthritis. This review summarizes recent findings on the regulation of hypertrophic chondrocyte differentiation, the cellular mechanisms involved in hypertrophy, and the role of chondrocyte hypertrophy in skeletal physiology and pathophysiology.

  5. Hypothyroidism - A cause for dilated cardiomyopathy in dogs; four year study (2008-2011

    Directory of Open Access Journals (Sweden)

    Satish Kumar Karlapudi

    Full Text Available Aim: The study was carried out to understand the thyroid dysfunction and its association with dilated cardiomyopathy in dogs. Materials and Methods: The study was done at Teaching Veterinary Hospital, Bhoiguda, College of Veterinary Science, Hyderabad for four years, i.e., from 2008 to 2011. A total of 256 dogs of various breed, age and sex were presented with typical skin and coat abnormalities. Few were also exhibiting signs of low metabolic rate. Skin sample analysis was done to rule out the causes of dermatitis. Thyroid profile was estimated to diagnose hypothyroidism. Selected cases were also subjected for echocardiography to study the association of cardiomyopathy. Based on thyroid profile, hypothyroid dogs were treated with levothyroxine @20mcg/kg wt, once daily, orally on empty stomach and dilated cardiomyopathy (DCM associated patients were additionally supplemented with enalapril @0.5 mg/kg, twice daily, orally for 6 months. The hemato biochemical and echocardiographic aspects are discussed. Results: The classical signs that were recorded in almost all the thyroid dysfunction dogs (231 were bilateral alopecia, rat tail and pigmentation and whereas, dyspnoea at rest, exercise intolerance, obesity, pale mucosae and corneal lipidosis were the significant low metabolic rate signs noticed in 42 dogs. However, syncope and seizures were also recorded in 31 of these hypothyroid dogs. Echocardiographic evaluation revealed significantly (P<0.01 increased LVEDd and LVEDs along with decreased IVS and LVPW both at systole and diastole among 33 dogs. Normocytic, normochromic and non-regenerative anemia and significantly (P<0.05 low T3, fT4, tT4 along with elevated serum cholesterol, triglycerides, TSH, CKMB, LDH and ALP were the hemato – biochemical findings among these dogs. After the initiation of therapy, improvement in clinical signs was noticed from day 7 and complete clinical recovery by the end of therapy. However, a non significant

  6. A Four-Year Longitudinal Study on Restless Legs Syndrome in Parkinson Disease

    Science.gov (United States)

    Moccia, Marcello; Erro, Roberto; Picillo, Marina; Santangelo, Gabriella; Spina, Emanuele; Allocca, Roberto; Longo, Katia; Amboni, Marianna; Palladino, Raffaele; Assante, Roberta; Pappatà, Sabina; Pellecchia, Maria Teresa; Barone, Paolo; Vitale, Carmine

    2016-01-01

    dopaminergic pathways, and worse sleep and cardiovascular disturbances. Citation: Moccia M, Erro R, Picillo M, Santangelo G, Spina E, Allocca R, Longo K, Amboni M, Palladino R, Assante R, Pappatà S, Pellecchia MT, Barone P, Vitale C. A four-year longitudinal study on restless legs syndrome in Parkinson disease. SLEEP 2016;39(2):405–412. PMID:26564123

  7. Malaria Four-year Epidemiological Trends in Sistan and Baluchistan Province, Iran

    Science.gov (United States)

    Norouzinezhad, Faezeh; Ghaffari, Fatemeh; Raeisi, Ahmad; Norouzinejad, Abbas; Kaveh, Farzad

    2017-01-01

    Introduction Malaria is one of the foremost public health concerns in Iran, where more than 90% of malaria cases are reported in the southern and south-eastern areas of the country. The aim of this study was to assess the epidemiological trends of malaria over a four-year period in in the Sistan and Baluchistan province in south east of Iran. Methods This descriptive epidemiological study examined malaria trends in Sistan and Baluchistan province from 2011 to 2014. The study used data collected in accordance with the Iranian Ministry of Health’s malaria control and elimination protocol. This protocol has digitized the data reporting system for malaria, and all information were sent online to the Center of Disease Control in the Ministry of Health. In this manner, information on malaria cases in Sistan and Baluchistan were made available for the researchers to analyze. Descriptive and comparative analyses were conducted using the SPSS version 13. Results Annual incidence rates reported in 2011, 2012, 2013, and 2014 showed the prevalence of 89.9, 43.9, 38.3 and 36.6 (per 100,000 persons), respectively. Across all 4 years, the highest numbers of cases were found in persons 16–25 years old and among males. Most of the infected individuals were villagers and workers. In total, 64.8% of patients were Iranian and 29.5% were Pakistani. The highest number of cases was diagnosed in the cities of Sarbaz and Chabahar, with 1,742 and 1,707 cases, respectively. The results showed that over the last 4 years, 50.8% of cases have entered into Iran from foreign countries. The majority of cases involved parasites in the trophozoite stage of the life cycle. In terms of surveillance, passive care was reported in the majority of cases, and vivax malaria had the highest prevalence in comparison with other types. Conclusion The findings are showing that the care, control and treatment system applied to Sistan and Baluchistan province has had a positive effect on decreasing the

  8. Experimental study of tissue-engineered cartilage allograft with RNAi chondrocytes in vivo

    Directory of Open Access Journals (Sweden)

    Wang ZH

    2014-05-01

    Full Text Available Zhenghui Wang,1 Xiaoli Li,2 Xi-Jing He,3 Xianghong Zhang,1 Zhuangqun Yang,4 Min Xu,1 Baojun Wu,1 Junbo Tu,5 Huanan Luo,1 Jing Yan11Department of Otolaryngology – Head and Neck Surgery, 2Department of Dermatology, 3Department of Orthopedics, The Second Hospital, Xi’an Jiaotong University, 4Department of Plastic and Burns Surgery, The First Hospital, Xi’an Jiaotong University, 5Department of Oral and Maxillofacial Plastic Surgery, The Stomatological Hospital, Xi’an Jiaotong University, Xi’an, People’s Republic of ChinaPurpose: To determine the effects of RNA interference (RNAi on chondrocyte proliferation, function, and immunological rejection after allogenic tissue-engineered cartilage transplantation within bone matrix gelatin scaffolds.Methods: Seven million rat normal and RNAi chondrocytes were harvested and separately composited with fibrin glue to make the cell suspension, and then transplanted subcutaneously into the back of Sprague Dawley rats after being cultured for 10 days in vitro. Untransplanted animals served as the control group. The allograft and immunological response were examined at 1, 2, 4, 8, and 12 months postoperatively with hematoxylin and eosin histochemical staining, immunohistochemical staining (aggrecan, type II collagen, class I and II major histocompatibility complex, and flow cytometry for peripheral blood cluster of differentiation 4+ (CD4+ and CD8+ T-cells.Results: There was no infection or death in the rats except one, which died in the first week. Compared to the control group, the RNAi group had fewer eukomonocytes infiltrated, which were only distributed around the graft. The ratio of CD4+/CD8+ T-cells in the RNAi group was significantly lower than the normal one (P<0.05. There were many more positively stained chondrocytes and positively stained areas around the cells in the RNAi group, which were not found in the control group.Conclusion: The aggrecanase-1 and aggrecanase-2 RNAi for chondrocytes

  9. Chondrocytes from congenital microtia possess an inferior capacity for in vivo cartilage regeneration to healthy ear chondrocytes.

    Science.gov (United States)

    Gu, Yunpeng; Kang, Ning; Dong, Ping; Liu, Xia; Wang, Qian; Fu, Xin; Yan, Li; Jiang, Haiyue; Cao, Yilin; Xiao, Ran

    2016-11-15

    The remnant auricular cartilage from microtia has become a valuable cell source for ear regeneration. It is important to clarify the issue of whether the genetically defective microtia chondrocytes could engineer cartilage tissue comparable to healthy ear chondrocytes. In the current study, the histology and cell yield of native microtia and normal ear cartilage were investigated, and the biological characteristics of derived chondrocytes examined, including proliferation, chondrogenic phenotype and cell migration. Furthermore, the in vivo cartilage-forming capacity of passaged microtia and normal auricular chondrocytes were systematically compared by seeding them onto polyglycolic acid/polylactic acid scaffold to generate tissue engineered cartilage in nude mice. Through histological examinations and quantitative analysis of glycosaminoglycan, Young's modulus, and the expression of cartilage-related genes, it was found that microtia chondrocytes had a slower dedifferentiation rate with the decreased expression of stemness-related genes, and weaker migration ability than normal ear chondrocytes, and the microtia chondrocytes-engineered cartilage was biochemically and biomechanically inferior to that constructed using normal ear chondrocytes. This study provides valuable information for the clinical application of the chondrocytes derived from congenital microtia to engineer cartilage. Copyright © 2016 John Wiley & Sons, Ltd.

  10. Four-year clinical outcome in asymptomatic patients undergoing coronary computed tomography angiography

    Institute of Scientific and Technical Information of China (English)

    ZHANG Kai-yi; GAI Lu-yue; GAI Jing-jing; HE Bin; GUAN Zhi-wei

    2013-01-01

    Background Percutaneous coronary intervention (PCI) is indicated for angina with coronary stenosis.However,PCI for asymptomatic coronary stenosis remains controversial.We prospectively followed a group of patients for four years who underwent coronary computed tomography angiography (CCTA) for major adverse cardiac events (MACE).We hypothesized that the results of this trial would reliably reflect the natural outcome of the coronary disease.Methods Consecutive patients who underwent CCTA from June 2008 to May 2009 were selected.Those who could not be reached by telephone,had significant angina,had CT images that were not interpretable,or poor kidney and left ventricular (LV) function were excluded.The patients were divided into five groups:group A normal CCTA without stenosis,group B mild stenosis (1%-49%),group C moderate stenosis (50%-74%),group D severe stenosis (≥75%) and they were treated with optimal medical therapy (OMT) or PCI.The group E had PCI before the CCTA examination.The patients were then followed for MACE after different treatments.MACE included acute myocardial infarction (MI),heart failure (HF) and death.Results The patient population consisted of 419 patients.The follow-up time was (51±5) months.The age was (60±31)years.Male made up 67.78% of the population (n=284).A total of 51 cases of MACE occurred including 25 MI,eight HF and 18 all-cause deaths.There was no MACE in group A.Although MACE occurred in two patients in group B,they were not attributed to cardiac death.We further compared the MACE in groups C-E and no significant difference was found (P >0.05).However,a difference was detected among patients with unstable angina pectoris (UAP),stable angina pectoris (SAP),re-hospitalization,and cerebrovascular events from groups A-E (P <0.05).The plaque scores were used to predict MACE.The scores progressively increased significantly with lesion severity (P <0.05).Receiver operating curve (ROC) was performed to determine the

  11. A Measurement of Electron Neutrino Appearance in the MINOS Experiment After Four Years of Data

    Energy Technology Data Exchange (ETDEWEB)

    Cavanaugh, Steven [Harvard Univ., Cambridge, MA (United States)

    2010-05-01

    experiment. After two years of running with 3.14 x 1020 protons-on-target (POT), a limit was set at δCP = 0 for the normal (inverted) hierarchy of sin2(2θ13) < 0.29 (0.42) at 90% C.L. This study finds after four years of data with an accumulated intensity of 7 x 1020 POT that sin2(2θ13) < 0.12 (0.20) with δCP = 0 at 90% C.L. for the normal (inverted) hierarchy.

  12. Subjective memory complaints in primary care patients and death from all causes: a four-year follow-up

    DEFF Research Database (Denmark)

    Siersma, Volkert Dirk; Waldemar, Gunhild; Waldorff, Frans Boch

    2013-01-01

    -related quality of life were collected at the enrolment primary care consultation. Dates of death from all causes occurring during the four-year follow-up were retrieved from the national databases. Cox proportional hazard regression models on time to death (censored after four years) were used to examine......OBJECTIVE: To investigate the prognostic value of subjective memory complaints for all-cause mortality in order to determine whether elderly persons with subjective memory complaints may be regarded as a group of vulnerable patients who need close follow-up. DESIGN: Prospective cohort study...... the influence of subjective memory complaints on all-cause mortality. RESULTS: 88 patients died during the four-year follow-up. The association between subjective memory complaints and mortality had a statistically not significant hazard ratio (HR) of 0.91, adjusting for known confounders. Statistically...

  13. Chondrocyte-specific ablation of Osterix leads to impaired endochondral ossification

    Energy Technology Data Exchange (ETDEWEB)

    Oh, Jung-Hoon [Department of Molecular Medicine, Cell and Matrix Research Institute, BK21 Medical Education Program for Human Resources, Kyungpook National University School of Medicine, Daegu (Korea, Republic of); Park, Seung-Yoon [Department of Biochemistry, School of Medicine, Dongguk University, Gyeongju 780-714 (Korea, Republic of); Crombrugghe, Benoit de [Department of Genetics, University of Texas, M.D. Anderson Cancer Center, Houston (United States); Kim, Jung-Eun, E-mail: kjeun@knu.ac.kr [Department of Molecular Medicine, Cell and Matrix Research Institute, BK21 Medical Education Program for Human Resources, Kyungpook National University School of Medicine, Daegu (Korea, Republic of)

    2012-02-24

    Highlights: Black-Right-Pointing-Pointer Conditional ablation of Osterix (Osx) in chondrocytes leads to skeletal defects. Black-Right-Pointing-Pointer Osx regulates chondrocyte differentiation and bone growth in growth plate chondrocytes. Black-Right-Pointing-Pointer Osx has an autonomous function in chondrocytes during endochondral ossification. -- Abstract: Osterix (Osx) is an essential transcription factor required for osteoblast differentiation during both intramembranous and endochondral ossification. Endochondral ossification, a process in which bone formation initiates from a cartilage intermediate, is crucial for skeletal development and growth. Osx is expressed in differentiating chondrocytes as well as osteoblasts during mouse development, but its role in chondrocytes has not been well studied. Here, the in vivo function of Osx in chondrocytes was examined in a chondrocyte-specific Osx conditional knockout model using Col2a1-Cre. Chondrocyte-specific Osx deficiency resulted in a weak and bent skeleton which was evident in newborn by radiographic analysis and skeletal preparation. To further understand the skeletal deformity of the chondrocyte-specific Osx conditional knockout, histological analysis was performed on developing long bones during embryogenesis. Hypertrophic chondrocytes were expanded, the formation of bone trabeculae and marrow cavities was remarkably delayed, and subsequent skeletal growth was reduced. The expression of several chondrocyte differentiation markers was reduced, indicating the impairment of chondrocyte differentiation and endochondral ossification in the chondrocyte-specific Osx conditional knockout. Taken together, Osx regulates chondrocyte differentiation and bone growth in growth plate chondrocytes, suggesting an autonomous function of Osx in chondrocytes during endochondral ossification.

  14. Applications of Chondrocyte-Based Cartilage Engineering: An Overview

    Science.gov (United States)

    Eo, Seong-Hui; Abbas, Qamar; Ahmed, Madiha

    2016-01-01

    Chondrocytes are the exclusive cells residing in cartilage and maintain the functionality of cartilage tissue. Series of biocomponents such as different growth factors, cytokines, and transcriptional factors regulate the mesenchymal stem cells (MSCs) differentiation to chondrocytes. The number of chondrocytes and dedifferentiation are the key limitations in subsequent clinical application of the chondrocytes. Different culture methods are being developed to overcome such issues. Using tissue engineering and cell based approaches, chondrocytes offer prominent therapeutic option specifically in orthopedics for cartilage repair and to treat ailments such as tracheal defects, facial reconstruction, and urinary incontinence. Matrix-assisted autologous chondrocyte transplantation/implantation is an improved version of traditional autologous chondrocyte transplantation (ACT) method. An increasing number of studies show the clinical significance of this technique for the chondral lesions treatment. Literature survey was carried out to address clinical and functional findings by using various ACT procedures. The current study was conducted to study the pharmacological significance and biomedical application of chondrocytes. Furthermore, it is inferred from the present study that long term follow-up studies are required to evaluate the potential of these methods and specific positive outcomes. PMID:27631002

  15. Fibroblast-like synoviocyte-chondrocyte interaction in cartilage degradation

    NARCIS (Netherlands)

    Steenvoorden, M.M.C.; Bank, R.A.; Ronday, H.K.; Toes, R.E.M.; Huizinga, T.W.J.; Groot, J. de

    2007-01-01

    Objective: In vitro models for joint diseases often focus on a single cell type, such as chondrocytes in osteoarthritis (OA) or fibroblast-like synoviocytes (synoviocytes) in rheumatoid arthritis (RA). However, these joint diseases affect the whole joint and interaction between chondrocytes and syno

  16. Applications of Chondrocyte-Based Cartilage Engineering: An Overview

    Directory of Open Access Journals (Sweden)

    Abdul-Rehman Phull

    2016-01-01

    Full Text Available Chondrocytes are the exclusive cells residing in cartilage and maintain the functionality of cartilage tissue. Series of biocomponents such as different growth factors, cytokines, and transcriptional factors regulate the mesenchymal stem cells (MSCs differentiation to chondrocytes. The number of chondrocytes and dedifferentiation are the key limitations in subsequent clinical application of the chondrocytes. Different culture methods are being developed to overcome such issues. Using tissue engineering and cell based approaches, chondrocytes offer prominent therapeutic option specifically in orthopedics for cartilage repair and to treat ailments such as tracheal defects, facial reconstruction, and urinary incontinence. Matrix-assisted autologous chondrocyte transplantation/implantation is an improved version of traditional autologous chondrocyte transplantation (ACT method. An increasing number of studies show the clinical significance of this technique for the chondral lesions treatment. Literature survey was carried out to address clinical and functional findings by using various ACT procedures. The current study was conducted to study the pharmacological significance and biomedical application of chondrocytes. Furthermore, it is inferred from the present study that long term follow-up studies are required to evaluate the potential of these methods and specific positive outcomes.

  17. Expansion of human nasal chondrocytes on macroporous microcarriers enhances redifferentiation

    NARCIS (Netherlands)

    Malda, J.; Kreijveld, E.; Temenoff, J.S.; Blitterswijk, van C.A.; Riesle, J.

    2003-01-01

    Articular cartilage has a limited capacity for self-repair. To overcome this problem, it is expected that functional cartilage replacements can be created from expanded chondrocytes seeded in biodegradable scaffolds. Expansion of chondrocytes in two-dimensional culture systems often results in dedif

  18. Expansion of human nasal chondrocytes on macroporous macrocarriers enhances redifferentiation

    NARCIS (Netherlands)

    Malda, J.; Kreijveld, E.; Temenoff, J.; Blitterswijk, van C.A.; Riesle, J.

    2003-01-01

    Articular cartilage has a limited capacity for self-repair. To overcome this problem, it is expected that functional cartilage replacements can be created from expanded chondrocytes seeded in biodegradable scaffolds. Expansion of chondrocytes in two-dimensional culture systems often results in dedif

  19. Dicer-dependent pathways regulate chondrocyte proliferation and differentiation.

    Science.gov (United States)

    Kobayashi, Tatsuya; Lu, Jun; Cobb, Bradley S; Rodda, Stephen J; McMahon, Andrew P; Schipani, Ernestina; Merkenschlager, Matthias; Kronenberg, Henry M

    2008-02-12

    Small noncoding RNAs, microRNAs (miRNAs), bind to messenger RNAs through base pairing to suppress gene expression. Despite accumulating evidence that miRNAs play critical roles in various biological processes across diverse organisms, their roles in mammalian skeletal development have not been demonstrated. Here, we show that Dicer, an essential component for biogenesis of miRNAs, is essential for normal skeletal development. Dicer-null growth plates show a progressive reduction in the proliferating pool of chondrocytes, leading to severe skeletal growth defects and premature death of mice. The reduction of proliferating chondrocytes in Dicer-null growth plates is caused by two distinct mechanisms: decreased chondrocyte proliferation and accelerated differentiation into postmitotic hypertrophic chondrocytes. These defects appear to be caused by mechanisms downstream or independent of the Ihh-PTHrP signaling pathway, a pivotal signaling system that regulates chondrocyte proliferation and differentiation. Microarray analysis of Dicer-null chondrocytes showed limited expression changes in miRNA-target genes, suggesting that, in the majority of cases, chondrocytic miRNAs do not directly regulate target RNA abundance. Our results demonstrate the critical role of the Dicer-dependent pathway in the regulation of chondrocyte proliferation and differentiation during skeletal development.

  20. Chondrocyte behavior on nanostructured micropillar polypropylene and polystyrene surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Prittinen, Juha [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland); Jiang, Yu [Department of Chemistry, University of Eastern Finland, Joensuu (Finland); Ylärinne, Janne H. [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland); Pakkanen, Tapani A. [Department of Chemistry, University of Eastern Finland, Joensuu (Finland); Lammi, Mikko J., E-mail: mikko.lammi@uef.fi [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland); Qu, Chengjuan [Department of Applied Physics, University of Eastern Finland, Kuopio (Finland)

    2014-10-01

    This study was aimed to investigate whether patterned polypropylene (PP) or polystyrene (PS) could enhance the chondrocytes' extracellular matrix (ECM) production and phenotype maintenance. Bovine primary chondrocytes were cultured on smooth PP and PS, as well as on nanostructured micropillar PP (patterned PP) and PS (patterned PS) for 2 weeks. Subsequently, the samples were collected for fluorescein diacetate-based cell viability tests, for immunocytochemical assays of types I and II collagen, actin and vinculin, for scanning electronic microscopic analysis of cell morphology and distribution, and for gene expression assays of Sox9, aggrecan, procollagen α{sub 1}(II), procollagen α{sub 1}(X), and procollagen α{sub 2}(I) using quantitative RT-PCR assays. After two weeks of culture, the bovine primary chondrocytes had attached on both patterned PP and PS, while practically no adhesion was observed on smooth PP. However, the best adhesion of the cells was on smooth PS. The cells, which attached on patterned PP and PS surfaces synthesized types I and II collagen. The chondrocytes' morphology was extended, and an abundant ECM network formed around the attached chondrocytes on both patterned PP and PS. Upon passaging, no significant differences on the chondrocyte-specific gene expression were observed, although the highest expression level of aggrecan was observed on the patterned PS in passage 1 chondrocytes, and the expression level of procollagen α{sub 1}(II) appeared to decrease in passaged chondrocytes. However, the expressions of procollagen α{sub 2}(I) were increased in all passaged cell cultures. In conclusion, the bovine primary chondrocytes could be grown on patterned PS and PP surfaces, and they produced extracellular matrix network around the adhered cells. However, neither the patterned PS nor PP could prevent the dedifferentiation of chondrocytes. - Highlights: • Methods to avoid chondrocyte dedifferentiation would be useful for cartilage

  1. Practices of Early Childhood Development Practitioners for Poor and Vulnerable Children from Birth to Four Years in South Africa

    Science.gov (United States)

    Ebrahim, Hasina Banu; Killian, Bev; Rule, Peter

    2011-01-01

    This article explores the practices undertaken by early childhood development practitioners (ECDPs) to support poor and vulnerable children from birth to four years outside centre-based provision. The article draws on part of a UNICEF (United Nations International Children's Fund) commissioned evaluation on family and community-based ECD (Early…

  2. An Examination of Early Retirement Plans for Faculty at Four-Year Private Colleges in New England.

    Science.gov (United States)

    Kennedy, M. Patricia

    College faculty early retirement plans at four-year private colleges in New England are compared to determine requirements for an early retirement policy. Findings from survey responses from 82 colleges included: 20 of the colleges had an early retirement plan; 90% of schools with an early retirement plan had a tenure policy; 25% of the schools…

  3. An Academic Comparison of Third Grade Reading Scores: Reflecting the Impact of Four-Year-Old Kindergarten

    Science.gov (United States)

    Towns, Debra

    2013-01-01

    In Wisconsin, increased academic performance has been a general assumption supporting the institutionalization and public funding of early childhood instruction. The purpose of this study was to determine if there was a relationship between attendance in public four-year-old kindergarten and elementary level reading performance. This study…

  4. Top Gainers: Some Public Four-Year Colleges and Universities Make Big Improvements in Minority Graduation Rates. College Results Online

    Science.gov (United States)

    Engle, Jennifer; Theokas, Christina

    2010-01-01

    Each year, nearly 400,000 minority students enroll as freshmen in a four-year college, hoping to realize all of the opportunities that earning a bachelor's degree affords. Many arrive on campus having overcome underfunded high schools, an intimidating college-admissions process, and daunting financial circumstances. Yet despite their persistence…

  5. Prediction of Job Satisfaction Based on Workplace Facets for Adjunct Business Faculty at Four-Year Universities

    Science.gov (United States)

    Lewis, Vance Johnson

    2012-01-01

    The purpose of this study was to examine the job satisfaction of adjuncts in the curriculum area of business at four-year universities and to determine the roles that individual job facets play in creating overall job satisfaction. To explore which job facets and demographics predict job satisfaction for the population, participants were asked to…

  6. The Effects of Deep Approaches to Learning on Students' Need for Cognition over Four Years of College

    Science.gov (United States)

    Wang, Jui-Sheng

    2013-01-01

    This study examines the effect of deep approaches to learning on development of the inclination to inquire and lifelong learning over four years, as an essential graduated outcome that helps students face the challenges of a complex and rapidly changing world. Despite the importance of the inclination to inquire and lifelong learning, some…

  7. A comparison of four years of limnology in the Sawtooth Lakes with emphasis on fertilization in Redfish Lake

    Energy Technology Data Exchange (ETDEWEB)

    Budy, P.; Luecke, C.; Wurtsbaugh, W.A.

    1996-05-01

    Included in this section of the report on limnology of Lakes in the Snake River Plain are descriptions of four years of limnological sampling to compare inter and intra annual variability in lake productivity to evaluate potential rearing conditions for juvenile sockeyed salmon. Data was used to evaluate the effects of nutrient enhancement, annual weather patterns, and planktivore consumption on lake productivity.

  8. Using Time-on-Task Measurements to Understand Student Performance in a Physics Class: A Four-Year Study

    Science.gov (United States)

    Stewart, John; Stewart, Gay; Taylor, Jennifer

    2012-01-01

    Student use of out-of-class time was measured for four years in the introductory second-semester calculus-based physics course at the University of Arkansas. Two versions of the course were presented during the time of the measurement. In both versions, the total out-of-class time a student invested in the course explained less than 1% of the…

  9. In-School Work Experience and the Returns to Two-Year and Four-Year Colleges

    Science.gov (United States)

    Molitor, C.J.; Leigh, D.E.

    2005-01-01

    Building on the existing education literature, we address two interrelated questions. First, how sensitive are estimated returns to two-year and four-year colleges to the inclusion of in-school work experience? Second, do the estimated returns to in-school work experience vary by type of educational institution attended? Regarding the first…

  10. Preventing errors in administration of parenteral drugs: the results of a four-year national patient safety program.

    NARCIS (Netherlands)

    Blok, C. de; Schilp, J.; Wagner, C.

    2013-01-01

    Objectives: To evaluate the implementation of a four-year national patient safety program concerning the parenteral drug administration process in the Netherlands. Methods: Structuring the preparation and administration process of parenteral drugs reduces the number of medication errors. A nationwid

  11. Developments in Instruction in Selected Four Year College Reading Improvement Programs Throughout the United States, 1950-1974.

    Science.gov (United States)

    Howard, Virginia Pearl Hymes

    The purpose of this study was to examine the organization, program format and duration of college reading improvement instructional programs (CRIPs) in selected four-year American colleges and universities from 1950-1974, and to gather information which is vital to effective curricula and planning for higher education in the area of reading…

  12. What's Happened to the Price of College? Quality-Adjusted Net Price Indexes for Four Year Colleges

    Science.gov (United States)

    Schwartz, Amy Ellen; Scafidi, Benjamin

    2004-01-01

    Hedonic models of the price of college to construct quality-adjusted net price indexes for U.S. four-year colleges were estimated. A 22 percent decline in the estimated price index is reported by adjusting for financial aid, while quality adjusting results lead to a smaller decline, for academic years 1990-91 to 1994-95.

  13. Autologous chondrocyte implantation in children and adolescents

    DEFF Research Database (Denmark)

    Schmal, H; Pestka, J M; Salzmann, G

    2013-01-01

    PURPOSE: Autologous chondrocyte implantation (ACI) is a well-established treatment method for cartilage defects in knees. Age-related grouping was based on expression data of cartilage-specific markers. Specificities of ACI in the different populations were analysed. METHODS: Two hundred and sixt...... months after the operation. CONCLUSIONS: Age-related expression of cartilage-specific markers allows definition of adolescents in cartilage regenerating surgery. Chondromalacia in these patients is mainly caused by OCD or trauma. LEVEL OF EVIDENCE: Case series, Level IV....

  14. Stent graft placement for dysfunctional arteriovenous grafts

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, Gyeong Sik [Dept. of Radiology, CHA Bundang Medical Center, College of Medicine, CHA University, Seongnam (Korea, Republic of); Shin, Byung Seok; Ohm, Joon Young; Ahn, Moon Sang [Chungnam National University Hospital, Daejeon (Korea, Republic of)

    2015-07-15

    This study aimed to evaluate the usefulness and outcomes of stent graft use in dysfunctional arteriovenous grafts. Eleven patients who underwent stent graft placement for a dysfunctional hemodialysis graft were included in this retrospective study. Expanded polytetrafluoroethylene covered stent grafts were placed at the venous anastomosis site in case of pseudoaneurysm, venous laceration, elastic recoil or residual restenosis despite the repeated angioplasty. The patency of the arteriovenous graft was evaluated using Kaplan-Meier analysis. Primary and secondary mean patency was 363 days and 741 days. Primary patency at 3, 6, and 12 months was 82%, 73%, and 32%, respectively. Secondary patency at the 3, 6, 12, 24, and 36 months was improved to 91%, 82%, 82%, 50%, and 25%, respectively. Fractures of the stent graft were observed in 2 patients, but had no effect on the patency. Stent graft placement in dysfunctional arteriovenous graft is useful and effective in prolonging graft patency.

  15. Induced superficial chondrocyte death reduces catabolic cartilage damage in murine posttraumatic osteoarthritis.

    Science.gov (United States)

    Zhang, Minjie; Mani, Sriniwasan B; He, Yao; Hall, Amber M; Xu, Lin; Li, Yefu; Zurakowski, David; Jay, Gregory D; Warman, Matthew L

    2016-08-01

    Joints that have degenerated as a result of aging or injury contain dead chondrocytes and damaged cartilage. Some studies have suggested that chondrocyte death precedes cartilage damage, but how the loss of chondrocytes affects cartilage integrity is not clear. In this study, we examined whether chondrocyte death undermines cartilage integrity in aging and injury using a rapid 3D confocal cartilage imaging technique coupled with standard histology. We induced autonomous expression of diphtheria toxin to kill articular surface chondrocytes in mice and determined that chondrocyte death did not lead to cartilage damage. Moreover, cartilage damage after surgical destabilization of the medial meniscus of the knee was increased in mice with intact chondrocytes compared with animals whose chondrocytes had been killed, suggesting that chondrocyte death does not drive cartilage damage in response to injury. These data imply that chondrocyte catabolism, not death, contributes to articular cartilage damage following injury. Therefore, therapies targeted at reducing the catabolic phenotype may protect against degenerative joint disease.

  16. Serum-free media for articular chondrocytes in vitro expansion

    Institute of Scientific and Technical Information of China (English)

    SHAO Xin-xin; Neil A.Duncan; LIN Lin; FU Xin; ZHANG Ji-ying; YU Chang-long

    2013-01-01

    Background In vitro chondrocyte expansion is a major challenge in cell-based therapy for human articular cartilage repair.Classical culture conditions usually use animal serum as a medium supplement,which raises a number of undesirable questions.In the present study,two kinds of defined,serum-free media were developed to expand chondrocytes in monolayer culture for the purpose of cartilage tissue engineering.Methods Bovine chondrocytes were expanded in serum-free media supplemented with fibroblast growth factor-2 and platelet-derived growth factor or fibroblast growth factor-2 and insulin-like growth factor.Expansion culture in a conventional 10% fetal bovine serum (FBS) medium served as control.Fibronectin coating was used to help cell adhesion in serum-free medium.Next,in vitro three-dimensional pellet culture was used to evaluate the chondrocyte capacity.Cell pellets were expanded in different media to re-express the differentiated phenotype (re-differentiation) and to form cartilaginous tissue.The pellets were assessed by glycosaminoglycans contents,collagen II,collagen I and collagen X immunohistological staining.Results Chondrocytes cultured in serum-free media showed no proliferation difference than cells grown with 10% FBS medium.In addition,chondrocytes expanded in both serum-free media expressed more differentiated phenotypes at the end of monolayer culture,as indicated by higher gene expression ratios of collagen type Ⅱ to collagen type Ⅰ.Pellets derived from chondrocytes cultured in both serum-free media displayed comparable chondrogenic capacities to pellets from cells expanded in 10% FBS medium.Conclusion These findings provide alternative culture approaches for chondrocytes in vitro expansion,which may benefit the clinical use of autologous chondrocytes implantation.

  17. Chondrocyte-seeded type I/III collagen membrane for autologous chondrocyte transplantation

    DEFF Research Database (Denmark)

    Niemeyer, Philipp; Lenz, Philipp; Kreuz, Peter C;

    2010-01-01

    PURPOSE: We report the 2-year clinical results and identify prognostic factors in patients treated with autologous chondrocyte transplantation by use of a collagen membrane to seed the chondrocytes (ACT-CS). METHODS: This is a prospective study of 59 patients who were treated with ACT-CS...... Repair Society (ICRS) rating, the percentage of patients rated A (normal) and B (nearly normal) increased from 33.9% preoperatively to 92.5% at 24 months after ACT-CS. IKDC and Lysholm scores increased from 50.1 points (SD, 13.4) and 60.5 points (SD, 9.4), respectively, to 76.1 points (SD, 15.2) (P...... CS as a salvage procedure. The rate of failures in patients with isolated cartilage defects was 5.9%. CONCLUSIONS: ACT-CS...

  18. Comparison of performance in a four year graduate entry medical programme and a traditional five/six year programme.

    OpenAIRE

    Byrne, Annette T.; Arnett, Richard; Farrell, Tom; Sreenan, Seamus

    2014-01-01

    Background In 2006 the Royal College of Surgeons in Ireland, (RCSI), introduced the first four year Graduate Entry Programme (GEP) in medicine in Ireland in line with national policy to broaden access to medical education. One concern considered at the time, was whether the GEP students could be trained to the same standard as their undergraduate Direct Entry Programme (DEP, five/six year duration) counterparts in the shorter time frame. Since students from both cohorts undertake the same exa...

  19. Coping With Strain (CWS) course - its effects on depressive symptoms: A four-year longitudinal randomized controlled trial.

    Science.gov (United States)

    Saelid, Gry Anette; Czajkowski, Nikolai Olavi; Holte, Arne; Tambs, Kristian; Aarø, Leif Edvard

    2016-08-01

    The Coping With Strain (CWS) course is a modification of the Coping With Depression (CWD) course. CWD is by far the most studied psycho-educational intervention to reduce and prevent depression, but CWD has never been tested in a randomized controlled trial in the workplace. This study seeks to examine the extent to which CWS, on a short-term and a long-term basis, reduces depressive symptoms in employees. After advertising at workplaces, 119 employees were randomized into Intervention Group I (IG1), which immediately participated in CWS, or Intervention Group II (IG2), which functioned as a control group for six months until its participation in CWS. The follow up period lasted for four years in both IG1 and IG2. Linear mixed models were fitted to the data. Depressive symptoms were significantly reduced during the course. The reduction of depressive symptoms was maintained over a period of four years in both IG1 and IG2, although there is a slight increase towards the end of the follow-up period. CWS is effective in reducing depressive symptoms among employees. The effects are long lasting and may be maintained over a period of four years.

  20. Modifiable risk factors predicting major depressive disorder at four year follow-up: a decision tree approach

    Directory of Open Access Journals (Sweden)

    Christensen Helen

    2009-11-01

    Full Text Available Abstract Background Relative to physical health conditions such as cardiovascular disease, little is known about risk factors that predict the prevalence of depression. The present study investigates the expected effects of a reduction of these risks over time, using the decision tree method favoured in assessing cardiovascular disease risk. Methods The PATH through Life cohort was used for the study, comprising 2,105 20-24 year olds, 2,323 40-44 year olds and 2,177 60-64 year olds sampled from the community in the Canberra region, Australia. A decision tree methodology was used to predict the presence of major depressive disorder after four years of follow-up. The decision tree was compared with a logistic regression analysis using ROC curves. Results The decision tree was found to distinguish and delineate a wide range of risk profiles. Previous depressive symptoms were most highly predictive of depression after four years, however, modifiable risk factors such as substance use and employment status played significant roles in assessing the risk of depression. The decision tree was found to have better sensitivity and specificity than a logistic regression using identical predictors. Conclusion The decision tree method was useful in assessing the risk of major depressive disorder over four years. Application of the model to the development of a predictive tool for tailored interventions is discussed.

  1. Immortalization of human articular chondrocytes and induction of their phenotype

    Institute of Scientific and Technical Information of China (English)

    何清义; 李起鸿; 杨柳; 许建中

    2003-01-01

    Objective To immortalize human articular chondrocytes (HACs) using gene transfection and to maintain stable phenotype of transformed HACs after induction.Methods HACs were transfected with the retroviral vector pLXSN encoding human papillomavirus 16E7 (HPV16E7), and the transformed clones were sorted and proliferated. Karyotype analysis, clone forming tests and nude mice tumor forming tests were applied to check the characteristics of the transformation. Type Ⅱ collagen of transformed chondrocytes was inducted with free serum medium (FSM) supplemented with nutridoma-sp and ascorbate. Results Immortalized HACs were isolated with fifty passages achieved. The HPV16E7 transformed cells were confirmed to be benign. Induction of FSM with nutridoma-sp and ascorbate promoted type Ⅱ collagen of transformed chondrocytes to the high levels of normal chondrocytes. Conclusion HACs transformed with HPV16E7 survive for long periods in vitro, and type Ⅱ collagen can maintain stability after induction.

  2. Four years of crusade!

    Directory of Open Access Journals (Sweden)

    Adilson Beatriz

    2012-04-01

    Full Text Available Orbital - The Electronic Journal of Chemistry is entering its 4th year of existence which is a reason to celebrate. A great work has been devoted to preserve a scientific journal of international reputation. This work was not always without mistakes or any trouble. However, these very adversities stimulate additional efforts in order to overcome our errors and, continuing with our serious proposal of editing a journal which scientists from all over the world can submit online their original works manuscripts to be published in peer review and open-access style. Beginning this year, Orbital was classified by the Chemistry Coordination Area on the B5 rank of CAPES Qualis (Brazil and, leveled as B4 by Coordination Areas of Materials and Engineering II. These important details demonstrate that Orbital is progressively attaining the acknowledgment of the Brazilian scientific community. The recognition by CAPES Qualis is crucial as the system evaluates the generation of knowledge in the Brazilian universities and the method by which this same knowledge disseminated to society. Not all these were possible without endeavoring of editors and volunteer assistance of advisors from several national and international institutions. Recently, the young Professors Kleber Thiago de Oliveira (UFSCAR, Grégoire Demetz (USP-RP and Amilcar Machulek Júnior (UFMS have joined us to share the idea and philosophy of Orbital by accepting our invitation to be associated editors. The tasks and challenges are enormous and, we are sure that new participants will support us on the way to achieve indexation in new Databases systems as SCOPUS, ISI and Scielo. Consequently, the international success of this electronic journal will be assured. We sincerely hope that in the academic Brazilian scenario, Orbital can figure as a solid bridge to interchange real creative science that is happening in various locations around the world.

  3. Identification of the calcitonin receptor in osteoarthritic chondrocytes

    Directory of Open Access Journals (Sweden)

    Christensen Tjorbjoern

    2011-10-01

    Full Text Available Abstract Background Preclinical and clinical studies have shown that salmon calcitonin has cartilage protective effects in joint degenerative diseases, such as osteoarthritis (OA. However, the presence of the calcitonin receptor (CTR in articular cartilage chondrocytes is yet to be identified. In this study, we sought to further investigate the expression of the CTR in naïve human OA articular chondrocytes to gain further confirmation of the existents of the CTR in articular cartilage. Methods Total RNA was purified from primary chondrocytes from articular cartilage biopsies from four OA patients undergoing total knee replacement. High quality cDNA was produced using a dedicated reverse transcription polymerase chain reaction (RT-PCR protocol. From this a nested PCR assay amplifying the full coding region of the CTR mRNA was completed. Western blotting and immunohistochemistry were used to characterize CTR protein on protein level in chondrocytes. Results The full coding transcript of the CTR isoform 2 was identified in all four individuals. DNA sequencing revealed a number of allelic variants of the gene including two potentially novel polymorphisms: a frame shift mutation, +473del, producing a shorter form of the receptor protein, and a single nucleotide polymorphism in the 3' non coding region of the transcript, +1443 C>T. A 53 kDa protein band, consistent with non-glycosylated CTR isoform 2, was detected in chondrocytes with a similar size to that expressed in osteoclasts. Moreover the CTR was identified in the plasma membrane and the chondrocyte lacuna of both primary chondrocytes and OA cartilage section. Conclusions Human OA articular cartilage chondrocytes do indeed express the CTR, which makes the articular a pharmacological target of salmon calcitonin. In addition, the results support previous findings suggesting that calcitonin has a direct anabolic effect on articular cartilage.

  4. Spontaneous Redifferentiation of Dedifferentiated Human Articular Chondrocytes on Hydrogel Surfaces

    OpenAIRE

    2010-01-01

    Chondrocytes rapidly dedifferentiate into a more fibroblastic phenotype on a two-dimensional polystyrene substratum. This impedes fundamental research on these cells as well as their clinical application. This study investigated the redifferentiation behavior of dedifferentiated chondrocytes on a hydrogel substratum. Dedifferentiated normal human articular chondrocyte–knee (NHAC-kn) cells were released from the sixth-passage monolayer cultured on a polystyrene surface. These cells were then s...

  5. Linoleate impairs collagen synthesis in primary cultures of avian chondrocytes.

    Science.gov (United States)

    Watkins, B A; Xu, H; Turek, J J

    1996-06-01

    The effects of supplemental fatty acids, vitamin E (VIT E), and iron-induced oxidative stress on collagen synthesis, cellular injury, and lipid peroxidation were evaluated in primary cultures of avian epiphyseal chondrocytes. The treatments included oleic and linoleic acids (O or 50 microM) complexed with BSA and dl-alpha-tocopheryl acetate (VIT E at 0 or 100 microM). After 14 days of preculture, the chondrocytes were enriched with fatty acids for 8 days then cultured with VIT E for 2 days. The chondrocytes were then treated with ferrous sulfate (O or 20 microM) for 24 hr to induce oxidative stress. Collagen synthesis was the lowest and the activity of lactate dehydrogenase (LDH) was the highest in chondrocyte cultures treated with 50 microM linoleic acid and 0 VIT E. In contrast, VIT E supplemented at 100 microM partially restored collagen synthesis in the chondrocytes enriched with linoleic acid and lowered LDH activity in the media. The iron oxidative inducer significantly increased the values of thiobarbituric acid-reactive substances (TBARS) in the culture medium. The data showed that linoleic acid impaired chondrocyte cell function and caused cellular injury but that VIT E reversed these effects. Results from a previous study demonstrated that VIT E stimulated bone formation in chicks fed unsaturated fat, and the present findings in cultures of epiphyseal chondrocytes suggest that VIT E is important for chondrocyte function in the presence of polyunsaturated fatty acids. VIT E appears to be beneficial for growth cartilage biology and in optimizing bone growth.

  6. The properties of bioengineered chondrocyte sheets for cartilage regeneration

    Directory of Open Access Journals (Sweden)

    Ota Naoshi

    2009-03-01

    Full Text Available Abstract Background Although the clinical results of autologous chondrocyte implantation for articular cartilage defects have recently improved as a result of advanced techniques based on tissue engineering procedures, problems with cell handling and scaffold imperfections remain to be solved. A new cell-sheet technique has been developed, and is potentially able to overcome these obstacles. Chondrocyte sheets applicable to cartilage regeneration can be prepared with this cell-sheet technique using temperature-responsive culture dishes. However, for clinical application, it is necessary to evaluate the characteristics of the cells in these sheets and to identify their similarities to naive cartilage. Results The expression of SOX 9, collagen type 2, 27, integrin α10, and fibronectin genes in triple-layered chondrocyte sheets was significantly increased in comparison to those in conventional monolayer culture and in a single chondrocyte sheet, implying a nature similar to ordinary cartilage. In addition, immunohistochemistry demonstrated that collagen type II, fibronectin, and integrin α10 were present in the triple-layered chondrocyte sheets. Conclusion The results of this study indicate that these chondrocyte sheets with a consistent cartilaginous phenotype and adhesive properties may lead to a new strategy for cartilage regeneration.

  7. Effect of autophagy induced by dexamethasone on senescence in chondrocytes

    Science.gov (United States)

    Xue, Enxing; Zhang, Yu; Song, Bing; Xiao, Jun; Shi, Zhanjun

    2016-01-01

    The aim of the current study was to explore the effects of dexamethasone (DXM) on autophagy and senescence in chondrocytes. Collagen II and aggrecan were examined in normal chondrocytes isolated from Sprague-Dawley rats. Following stimulation with DXM, LysoTracker Red staining, monodansylcadaverine (MDC) staining, green fluorescent protein-red fluorescent protein-light chain 3 (LC3) and western blotting were used to detect autophagy levels in the chondrocytes. Mechanistic target of rapamycin (mTOR) pathway-associated molecules were investigated by western blotting. Cell senescence was analyzed by senescence-associated (SA)-β-galactosidase (β-gal) staining. A dose-dependent increase in the number of autophagic vacuoles was observed in the DXM-treated chondrocytes, as demonstrated by LysoTracker Red and MDC staining. A dose-dependent increase in autophagosome formation was observed in the DXM-treated chondrocytes. Expression of LC3-II and beclin-1 was increased by DXM, in particular in the cells treated with DXM for 4 days. However, P62 expression was reduced as a result of treatment. SA-β-gal staining indicated that DXM increased cell senescence. Notably, DXM-induced cell senescence was exacerbated by the autophagic inhibitor 3-MA. Autophagy induced by DXM protected chondrocytes from senescence, and it is suggested that the mTOR pathway may be involved in the activation of DXM-induced autophagy. PMID:27572674

  8. LOCAL INTERSTELLAR HYDROGEN'S DISAPPEARANCE AT 1 AU: FOUR YEARS OF IBEX IN THE RISING SOLAR CYCLE

    Energy Technology Data Exchange (ETDEWEB)

    Saul, Lukas; Rodriguez, Diego; Scheer, Juergen; Wurz, Peter [University of Bern, Bern (Switzerland); Bzowski, Maciej; Kubiak, Marzena; Sokol, Justina [Space Research Centre PAS, Warsaw (Poland); Fuselier, Stephen; McComas, Dave [Southwest Research Institute, San Antonio, TX (United States); Moebius, Eberhard [University of New Hampshire, Durham, NH (United States)

    2013-04-20

    NASA's Interstellar Boundary Explorer (IBEX) mission has recently opened a new window on the interstellar medium (ISM) by imaging neutral atoms. One ''bright'' feature in the sky is the interstellar wind flowing into the solar system. Composed of remnants of stellar explosions as well as primordial gas and plasma, the ISM is by no means uniform. The interaction of the local ISM with the solar wind shapes our heliospheric environment with hydrogen being the dominant component of the very local ISM. In this paper, we report on direct sampling of the neutral hydrogen of the local ISM over four years of IBEX observations. The hydrogen wind observed at 1 AU has decreased and nearly disappeared as the solar activity has increased over the last four years; the signal at 1 AU has dropped off in 2012 by a factor of {approx}8 to near background levels. The longitudinal offset has also increased with time presumably due to greater radiation pressure deflecting the interstellar wind. We present longitudinal and latitudinal arrival direction measurements of the bulk flow as measured over four years beginning at near solar minimum conditions. The H distribution we observe at 1 AU is expected to be different from that outside the heliopause due to ionization, photon pressure, gravity, and filtration by interactions with heliospheric plasma populations. These observations provide an important benchmark for modeling of the global heliospheric interaction. Based on these observations we suggest a further course of scientific action to observe neutral hydrogen over a full solar cycle with IBEX.

  9. Corneal endothelium after deep anterior lamellar keratoplasty and penetrating keratoplasty for keratoconus: A four-year comparative study

    Directory of Open Access Journals (Sweden)

    Anil Kubaloglu

    2012-01-01

    Full Text Available Purpose: To compare the status of corneal endothelium and central corneal thickness within the first four postoperative years after deep anterior lamellar keratoplasty (DALK and penetrating keratoplasty (PK in patients with keratoconus. Materials and Methods: Thirty-nine eyes (Group A which had PK and 44 eyes (Group B which had DALK for the treatment of keratoconus were included in this retrospective study. The endothelial cell density (ECD, the mean endothelial cell area and the coefficient of variation of cell area were assessed with a non-contact specular microscope, and the central corneal thickness (CCT was measured with an ultrasound pachymeter. Results: Mean ECD loss rate at two years was 36.24% in Group A and 18.12% in Group B (P<0.001. Mean ECD loss rate at four years was 47.82% in Group A and 21.62% in Group B (P<0.001. Mean annual ECD loss rate was calculated 14.12% per year in Group A and 5.78% per year in Group B. In the PK group, increase in mean CCT was 15.60% in two years and 15.03% in four years, while in the DALK group, mean CCT increased by 8.05% in two years and 9.31% in four years. Conclusions: As the majority of ectatic disorders such as keratoconus occur in young people, long-term endothelial cell survival following treatment with keratoplasty is essential for the long-term visual ability. Our finding that corneal endothelial cell loss in the DALK group occurs at a slower rate than in the PK group suggests DALK as a safer alternative to PK in these selected patients.

  10. Searches for Extended and Point-like Neutrino Sources with Four Years of IceCube Data

    DEFF Research Database (Denmark)

    Aartsen, M.G.; Ackermann, M.; Adams, J.

    2014-01-01

    We present results on searches for point-like sources of neutrinos using four years of IceCube data, including the first year of data from the completed 86 string detector. The total livetime of the combined data set is 1373 days. For an E –2 spectrum, the observed 90% C.L. flux upper limits are ...... of active galactic nuclei. In all cases, the data are compatible with the background-only hypothesis, and upper limits on the flux of muon neutrinos are reported for the sources considered....

  11. Using time-on-task measurements to understand student performance in a physics class: A four-year study

    OpenAIRE

    John Stewart; Gay Stewart†; Jennifer Taylor

    2012-01-01

    Student use of out-of-class time was measured for four years in the introductory second-semester calculus-based physics course at the University of Arkansas. Two versions of the course were presented during the time of the measurement. In both versions, the total out-of-class time a student invested in the course explained less than 1% of the variance in test average and in normalized gain on the Conceptual Survey in Electricity and Magnetism. The details of how students used their out-of-cla...

  12. Where to find a trustworthy source of information? four years of risk communication about a nuclear waste disposal

    Energy Technology Data Exchange (ETDEWEB)

    Findahl, O. [Dept. of Media and Communication, Umea University, Umea (Sweden)

    1998-07-01

    Full text of publication follows: in September 1997 there was a referendum, in a small local district in the north of Sweden, on the disposal of the swedish high intensive nuclear waste. A small majority of the 2500 voters stopped the examination of the conditions for a storage. Four years earlier the opinionagainst a local storage of nuclear waste had been much stronger but during the years that followed more people came to think that an examination of the conditions for a storage was OK; that will create new jobs, and what will come later we will see. The closer to the referendum the stronger this positive opinion became. They did not have any information problems. There were experts and information sources they could believe in. But the majority was still negative and they were dissatisfied. They could not find any trustworthy information. And though they in the end had prevented the localization of the nuclear waste storage to their commune they still felt that the decision had been taken high above their heads. Why is it like that? And what happened during these four years of experimentation in democratic risk communication? (author)

  13. Extension of general practice training from three to four years: experiences of a vocational training programme in Southern Ireland.

    LENUS (Irish Health Repository)

    Dowling, Stephanie

    2009-05-01

    The aim of this study was to evaluate the experiences of trainees taking part in an extended (four-year) general practice training programme introduced in the South Eastern region of the Republic of Ireland to replace the previous traditional (three-year) programme. In a qualitative design, eight homogeneous focus groups were held to determine the value of the additional year of training. The first cohort of trainees was interviewed towards the start and at the end of their fourth year. Trainees finishing the following year were also interviewed, as were graduates from the final three-year programme. GP trainers and the four members of the programme directing team comprised two further independent focus groups. Trainees reported that the integration of hospital posts and general practice attachments over the four years was particularly beneficial. The exposure to a variety of different general practices and the opportunity to take part in specialty clinics were considered extremely useful. The fourth year of training was felt to be less pressurised than previous years. Professional and personal development was enhanced; improved readiness to practise and confidence were noted. Perceived disadvantages of extended training included a lack of acknowledgment for doctors in their fourth year and excessive emphasis placed on research during the final year of training. The addition of an extra year of vocational training improves professional and personal development and changes the learning experience for doctors. Doctors felt more confident and ready to enter independent practice at the end of the fourth year of training.

  14. Low oxygen tension stimulates redifferentiation of dedifferentiated adult human nasal chondrocytes

    NARCIS (Netherlands)

    Malda, J.; Blitterswijk, van C.A.; Geffen, van M.; Martens, D.E.; Tramper, J.; Riesle, J.

    2004-01-01

    Objective: To determine the effect of dissolved oxygen tension (DO) on the redifferentiation of dedifferentiated adult human nasal septum chondrocytes cultured as pellets. Design: After isolation, human nasal chondrocytes were expanded in monolayer culture, which resulted in their dedifferentiation.

  15. Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes

    NARCIS (Netherlands)

    Malda, J.; Blitterswijk, van C.A.; Geffen, van M.; Martens, D.E.; Tramper, J.; Riesle, J.

    2004-01-01

    Objective: To determine the effect of dissolved oxygen tension (DO) on the redifferentiation of dedifferentiated adult human nasal septum chondrocytes cultured as pellets. - Design: After isolation, human nasal chondrocytes were expanded in monolayer culture, which resulted in their dedifferentiati

  16. Extracellular matrix domain formation as an indicator of chondrocyte dedifferentiation and hypertrophy

    NARCIS (Netherlands)

    Wu, Ling; Gonzalez, Stephanie; Shah, Saumya; Kyupelyan, Levon; Petrigliano, Frank A.; McAllister, David R.; Adams, John S.; Karperien, Marcel; Tuan, Tai-Lan; Benya, Paul D.; Evseenko, Denis

    2014-01-01

    Cartilage injury represents one of the most significant clinical conditions. Implantation of expanded autologous chondrocytes from noninjured compartments of the joint is a typical strategy for repairing cartilage. However, two-dimensional culture causes dedifferentiation of chondrocytes, making the

  17. Bone grafting: An overview

    Directory of Open Access Journals (Sweden)

    D. O. Joshi

    2010-08-01

    Full Text Available Bone grafting is the process by which bone is transferred from a source (donor to site (recipient. Due to trauma from accidents by speedy vehicles, falling down from height or gunshot injury particularly in human being, acquired or developmental diseases like rickets, congenital defects like abnormal bone development, wearing out because of age and overuse; lead to bone loss and to replace the loss we need the bone grafting. Osteogenesis, osteoinduction, osteoconduction, mechanical supports are the four basic mechanisms of bone graft. Bone graft can be harvested from the iliac crest, proximal tibia, proximal humerus, proximal femur, ribs and sternum. An ideal bone graft material is biologically inert, source of osteogenic, act as a mechanical support, readily available, easily adaptable in terms of size, shape, length and replaced by the host bone. Except blood, bone is grafted with greater frequency. Bone graft indicated for variety of orthopedic abnormalities, comminuted fractures, delayed unions, non-unions, arthrodesis and osteomyelitis. Bone graft can be harvested from the iliac crest, proximal tibia, proximal humerus, proximal femur, ribs and sternum. By adopting different procedure of graft preservation its antigenicity can be minimized. The concept of bone banking for obtaining bone grafts and implants is very useful for clinical application. Absolute stability require for successful incorporation. Ideal bone graft must possess osteogenic, osteoinductive and osteocon-ductive properties. Cancellous bone graft is superior to cortical bone graft. Usually autologous cancellous bone graft are used as fresh grafts where as allografts are employed as an alloimplant. None of the available type of bone grafts possesses all these properties therefore, a single type of graft cannot be recomm-ended for all types of orthopedic abnormalities. Bone grafts and implants can be selected as per clinical problems, the equipments available and preference of

  18. The national neonatal screening programme for congenital toxoplasmosis in Denmark: results from the initial four years, 1999-2002

    DEFF Research Database (Denmark)

    Schmidt, Dorte Remmer; Høgh, Birthe; Andersen, O;

    2006-01-01

    AIMS: To describe the outcome of four years' nationwide neonatal screening for congenital toxoplasmosis in liveborn newborns. METHODS: Congenital toxoplasmosis was diagnosed if specific Toxoplasma gondii IgM antibodies were detected in eluate from the PKU Guthrie filter paper card from a child....... Infants diagnosed with congenital toxoplasmosis were examined for intracranial and retinal lesions and treated for three months with sulphadiazine, pyrimethamine, and folinic acid continuously. RESULTS: Eluates from PKU-cards from 262 912 newborns were analysed. The birth prevalence of congenital...... toxoplasma infection was 2.1 per 10 000 liveborns. Congenital toxoplasmosis was suspected in 96 infants and confirmed in 55. Forty seven children were examined for intracranial and retinal lesions soon after birth; 12 had clinical signs at this first examination. Of these, 5 had intracranial calcifications...

  19. Recovery of a subtropical rocky shore is not yet complete, four years after a moderate sized oil spill.

    Science.gov (United States)

    Finlayson, Kimberly; Stevens, Tim; Arthur, James Michael; Rissik, David

    2015-04-15

    Little is known about the recovery trajectory from small to moderate spills (rocky shores. This study examines the recovery of the rocky shore community four years after the spill. Results indicate that recovery on Moreton Island is taking longer than the 3-4 years suggested by the literature. The upper shore is recovering faster than the mid shore and is nearly recovered while the mid shore is still in the recovery process. These results indicate that small to moderate sized spills can have environmental impacts on par with much larger spills and emphasizes the need for a clear definition of a recovery endpoint. Long term studies are required to gain a full understanding of trajectories of recovery after oil spill impacts.

  20. Mammary tumors and serum hormones in the bitch treated with medroxyprogesterone acetate or progesterone for four years

    Energy Technology Data Exchange (ETDEWEB)

    Frank, D.W.; Kirton, K.T.; Murchison, T.E.; Quinlan, W.J.; Coleman, M.E.; Gilbertson, T.J.; Feenstra, E.S.; Kimball, F.A.

    1978-01-01

    After four years of a long term contraceptive steroid safety study, the incidence and the histologic type of mammary dysplasia produced is similar in beagles treated with medroxyprogesterone acetate (medroxyprogesterone) or progesterone. Serum insulin, thyroid stimulating hormone (TSH), triiodothyronine, growth hormone, prolactin, 17..beta..-estradiol, progesterone, and cortisol were determined by radioimmunoassay on samples collected after 45 months of treatment. Serum growth hormone and insulin concentrations were elevated in a dose related manner in both treatment groups. Triiodothyronine, cortisol, and estradiol-17..beta.. (medroxyprogesterone only) were lowered. TSH and prolactin concentrations were not changed. Pituitary--gonadal hormone interaction in the pathogenesis of mammary neoplasia of the dog is discussed. Prolonged treatment of the beagle with massive doses of progesterone or medroxyprogesterone results in a dose related incidence of mammary modules.

  1. Species Diversity Effects on Productivity, Persistence and Quality of Multispecies Swards in a Four-Year Experiment

    Science.gov (United States)

    Søegaard, Karen; Cong, Wen-Feng; Eriksen, Jørgen

    2017-01-01

    Plant species diversity may benefit natural grassland productivity, but its effect in managed grassland systems is not well understood. A four-year multispecies grassland experiment was conducted to investigate the effect of species diversity–legumes and non-leguminous forbs–on productivity, persistence and sward quality under cutting or grazing regimes and with or without slurry application. Three mixtures were established– 3-mix: grass, red and white clover, 10-mix: 3-mix plus birdsfoot trefoil and six non-leguminous forbs, and 12-mix: 10-mix plus lucerne and festulolium. Species diversity increased sward production and yield persistence under cutting regime. The 12-mix had the highest yield from the second year onwards and no statistically significant yield reduction over four years, while annual yields in the 3-mix and 10-mix decreased significantly with increasing grassland age. The higher yield in the 12-mix was mainly due to the inclusion of high-yielding lucerne. The 10-mix and 12-mix had lower proportions of unsown species than the 3-mix, the difference being dependent on grassland age. Generally, the 3-mix had higher concentrations of in-vitro organic matter digestibility (IVOMD), neutral detergent fiber (NDF) and crude protein (CP), and a lower concentration of ash than the 10-mix and 12-mix. Slurry application increased annual yield production by 10% and changed the botanical composition, increasing the proportion of grass and decreasing the proportion of legumes. Compared to cutting, grazing increased forage production by 9% per cut on average and lowered legume and forb proportions in the mixtures, but yields did not differ among the three mixtures. Overall, our results suggest that species diversity increases sward productivity and persistence only under an ungrazed cutting regime. We conclude that increasing species diversity by selecting appropriate species with compatible management is key to achieving both high yields and high persistence

  2. ACL Revision in Synthetic ACL graft failure

    Science.gov (United States)

    Etcheto, H. Rivarola; Zordán, J.; Escobar, G.; Collazo, C.; Palanconi, M.; Autorino, C.; Salinas, E. Alvarez

    2017-01-01

    The development of synthetic grafts as an alternative to biological grafts for reconstruction of the anterior cruciate ligament dates from 1980. The interest is awakened due to the potential advantages of: The absence of morbidity associated with donor site, and early return to sport. However, this surgical technique has had multiple complications associated with graft: mechanical failures (synthetic graft failure, loss of fixation), synovial foreign body reaction, recurrent stroke, recurrent instability and ultimately, early osteoarthritis. Objectives: We describe the synthetic graft failure LCA, intraoperative findings and details of surgical technique. Methods: Patient 35 years old, with a history of ACL reconstruction four years of evolution in another health center, consultation with the Service knee arthroscopy for acute knee pain left knee during secondary sporting event to a rotation mechanism with fixed foot. On physical examination, presents and positive Lachman maneuver Pivot. Radiografia in a widening of the tibial tunnel is observed. NMR shows a discontinuity of fibers of synthetic graft. Results: First time arthroscopic revision where synthetic plastic LCA identifies with Disruption fiber pattern. Intraoperatively, hypertrophic chronic synovitis localized predominantly in intercóndilo is observed. debridement thereof is performed, and proceeds to the extraction of the synthetic ligament. Then he was made prior cruentado and revival of the edges of the tunnel, filling them with non-irradiated structural bone allograft. At four months as planned and after confirmation by studies incorporating bone graft was performed the second time with the new plastic ACL. It was planned like a primary graft surgery with autologous hamstring prepared in fourfold form, and fixation with modified transtibial technique Biotransfix system proximal and distal screw Biocomposite (arthrex®). A quadruple graft 9 mm was obtained, making good positioning of tunnels and stable

  3. Autophagy modulates articular cartilage vesicle formation in primary articular chondrocytes.

    Science.gov (United States)

    Rosenthal, Ann K; Gohr, Claudia M; Mitton-Fitzgerald, Elizabeth; Grewal, Rupinder; Ninomiya, James; Coyne, Carolyn B; Jackson, William T

    2015-05-22

    Chondrocyte-derived extracellular organelles known as articular cartilage vesicles (ACVs) participate in non-classical protein secretion, intercellular communication, and pathologic calcification. Factors affecting ACV formation and release remain poorly characterized; although in some cell types, the generation of extracellular vesicles is associated with up-regulation of autophagy. We sought to determine the role of autophagy in ACV production by primary articular chondrocytes. Using an innovative dynamic model with a light scatter nanoparticle counting apparatus, we determined the effects of autophagy modulators on ACV number and content in conditioned medium from normal adult porcine and human osteoarthritic chondrocytes. Healthy articular chondrocytes release ACVs into conditioned medium and show significant levels of ongoing autophagy. Rapamycin, which promotes autophagy, increased ACV numbers in a dose- and time-dependent manner associated with increased levels of autophagy markers and autophagosome formation. These effects were suppressed by pharmacologic autophagy inhibitors and short interfering RNA for ATG5. Caspase-3 inhibition and a Rho/ROCK inhibitor prevented rapamycin-induced increases in ACV number. Osteoarthritic chondrocytes, which are deficient in autophagy, did not increase ACV number in response to rapamycin. SMER28, which induces autophagy via an mTOR-independent mechanism, also increased ACV number. ACVs induced under all conditions had similar ecto-enzyme specific activities and types of RNA, and all ACVs contained LC3, an autophagosome-resident protein. These findings identify autophagy as a critical participant in ACV formation, and augment our understanding of ACVs in cartilage disease and repair.

  4. THE EFFECT OF PIROXICAM ON THE METABOLISM OF ISOLATED HUMAN CHONDROCYTES

    NARCIS (Netherlands)

    BULSTRA, SK; KUIJER, R; BUURMAN, WA; TERWINDTROUWENHORST, E; GUELEN, PJM; VANDERLINDEN, AJ

    1992-01-01

    The effect of piroxicam on the metabolism of healthy and osteoarthrotic (OA) chondrocytes was studied in vitro. The chondrocytes were obtained from five healthy, five moderately OA, and four severely OA hips or knees. The chondrocytes were cultured in a high-density, short-term in vitro model. In th

  5. File list: His.Bon.10.AllAg.Chondrocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  12. File list: ALL.Bon.20.AllAg.Chondrocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  18. File list: His.Bon.50.AllAg.Chondrocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  19. Cell manipulation in autologous chondrocyte implantation: from research to cleanroom.

    Science.gov (United States)

    Roseti, Livia; Serra, Marta; Tigani, Domenico; Brognara, Irene; Lopriore, Annamaria; Bassi, Alessandra; Fornasari, Pier Maria

    2008-04-01

    In the field of orthopaedics, autologous chondrocyte implantation is a technique currently used for the regeneration of damaged articular cartilage. There is evidence of the neo-formation of tissue displaying characteristics similar to hyaline cartilage. In vitro chondrocyte manipulation is a crucial phase of this therapeutic treatment consisting of different steps: cell isolation from a cartilage biopsy, expansion in monolayer culture and growth onto a three-dimensional biomaterial to implant in the damaged area. To minimise the risk of in vitro cell contamination, the manipulation must be performed in a controlled environment such as a cleanroom. Moreover, the choice of reagents and raw material suitable for clinical use in humans and the translation of research protocols into standardised production processes are important. In this study we describe the preliminary results obtained by the development of chondrocyte manipulation protocols (isolation and monolayer expansion) in cleanrooms for the application of autologous implantation.

  20. Astaxanthin reduces matrix metalloproteinase expression in human chondrocytes.

    Science.gov (United States)

    Chen, Wei-Ping; Xiong, Yan; Shi, Yong-Xiang; Hu, Peng-Fei; Bao, Jia-Peng; Wu, Li-Dong

    2014-03-01

    Astaxanthin is a red carotenoid pigment which exerts multiple biological activities. However, little is known about the effects of astaxanthin on matrix metalloproteinases (MMPs) in OA. The present study investigated the effects of astaxanthin on MMPs in human chondrocytes. Human chondrocytes were pretreated with astaxanthin at 1, 10 or 50μM, then, cells were stimulated with IL-1β (10ng/ml) for 24h. MMP-1, MMP-3 and MMP-13 were observed. We found that astaxanthin reduced the expression of MMP-1, MMP-3 and MMP-13 as well as the phosphorylation of two mitogen-activated protein kinases (MAPK) (p38 and ERK1/2) in IL-1β-stimulated chondrocytes. Astaxanthin also blocked the IκB-α degradation. These results suggest that astaxanthin may be beneficial in the treatment of OA.

  1. Epigenetic Regulation of Chondrocyte Catabolism and Anabolism in Osteoarthritis.

    Science.gov (United States)

    Kim, Hyeonkyeong; Kang, Donghyun; Cho, Yongsik; Kim, Jin-Hong

    2015-08-01

    Osteoarthritis (OA) is one of the most prevalent forms of joint disorder, associated with a tremendous socioeconomic burden worldwide. Various non-genetic and lifestyle-related factors such as aging and obesity have been recognized as major risk factors for OA, underscoring the potential role for epigenetic regulation in the pathogenesis of the disease. OA-associated epigenetic aberrations have been noted at the level of DNA methylation and histone modification in chondrocytes. These epigenetic regulations are implicated in driving an imbalance between the expression of catabolic and anabolic factors, leading eventually to osteoarthritic cartilage destruction. Cellular senescence and metabolic abnormalities driven by OA-associated risk factors appear to accompany epigenetic drifts in chondrocytes. Notably, molecular events associated with metabolic disorders influence epigenetic regulation in chondrocytes, supporting the notion that OA is a metabolic disease. Here, we review accumulating evidence supporting a role for epigenetics in the regulation of cartilage homeostasis and OA pathogenesis.

  2. The impact of tenofovir disoproxil fumarate on kidney function: four-year data from the HIV-infected outpatient cohort

    Directory of Open Access Journals (Sweden)

    Nadine Monteiro

    2014-11-01

    Full Text Available Introduction: With improvements in survival and disease progression in the era of combined antiretroviral therapy, complications such as kidney disease are becoming increasingly prevalent in HIV-infected patients. Tenofovir disoproxil fumarate (TDF has been associated with nephrotoxicity, including decline in glomerular filtration rate, proximal tubular damage and acute kidney injury. Objective: Characterize kidney safety of TDF-containing antiretroviral treatment (ART regimens in HIV-infected patients. Methods: Non-controlled, observational, retrospective study was based on the clinical files registry of HIV patients who started TDF between January and December 2008. We assessed outpatients followed at a single Portuguese center. Demographic, clinical, virological and immunological data at baseline were collected. Serum creatinine, estimated glomerular filtration rate (eGFR and creatinine clearance (CrCL were assessed at baseline, after six months and every year up to four years. CrCL and eGFR were calculated by Cockroft–Gault and Modification of Diet in Renal Disease equations, respectively. Results: A total of 176 patients (71.6% males with a mean age of 43 years were enrolled. Ninety-six (52% were ART-naive patients at TDF initiation. At baseline 12.5% had hypertension, 4% diabetes, 25% chronic hepatitis C and 9% chronic hepatitis B infections; 58% had normal renal function (eGFR ≥90 ml/min/1.73 m2, 36% had mild (eGFR 60-89 ml/min/1.73 m2 renal dysfunction and 2.3% had moderate (eGFR 30-59 ml/min/1.73 m2 renal dysfunction at initiation of TDF. Eighty-three (47% patients were on protease inhibitors and the remaining on NNRTIs containing regimens. During 48 months follow-up, 5% experienced moderate renal dysfunction and 1.7% severe renal dysfunction. Twenty-one (12% patients met the definition criteria of rapid decline of renal function (annual decline of eGFR ≥3 ml/min/1.73 m2 in two consecutive years. The development of kidney events

  3. Exploring the experiences of female students in introductory project-based engineering courses at two- and four-year institutions

    Science.gov (United States)

    Swan, Amy K.

    2011-12-01

    This qualitative study explored the experiential and contextual factors that shaped female students' pathways into introductory project-based engineering classes at two community colleges and one four-year institution, as well as female students' experiences within and outside of these classes. The study was framed by Social Cognitive Career Theory (SCCT) (Lent, Brown & Hackett, 1996) and Bronfenbrenner's (1979) ecological systems theory. Findings were based on analyses of data gathered through multiple methods: observations; individual interviews with female students; focus group interviews with project teams; and document collection. The findings of this study revealed that while positive experiences with math or science were a likely pre-cursor to engineering interest, experiential learning appeared to be a more powerful force in fostering students' engineering interest. Specifically, participants developed an interest in engineering through academic, professional, and extracurricular engineering- and design-related activities that familiarized them with the tasks and skills involved in engineering work and helped them develop a sense of selfefficacy with regard to this work. Interest and self-efficacy, in turn, played a role in students' postsecondary educational decision-making processes, as did contextual factors including families and finances. This study's findings also showed that participants' project teams were a critically important microsystem within participants' ecological environments. Within this sometimes "chilly" microsystem, female students negotiated intrateam processes, which were in some cases affected by gender norms. Intrateam processes that influenced female students' project-based learning experiences included: interpersonal dynamics; leadership; and division of labor. This study also identified several ways in which the lived experiences of participants at the community colleges were different from, or similar to, those of participants

  4. Efficacy and safety of early cyclosporine conversion to sirolimus with continued MMF-four-year results of the Postconcept study.

    Science.gov (United States)

    Lebranchu, Y; Thierry, A; Thervet, E; Büchler, M; Etienne, I; Westeel, P F; Hurault de Ligny, B; Moulin, B; Rérolle, J P; Frouget, T; Girardot-Seguin, S; Toupance, O

    2011-08-01

    Calcineurin inhibitor (CNI) withdrawal has been used as a strategy to improve renal allograft function. We previously reported that conversion from cyclosporine A (CsA) to sirolimus (SRL) 3 months after transplantation significantly improved renal function at 1 year. In the Postconcept trial, 77 patients in the SRL group and 85 in the CsA group were followed for 48 months. Renal function (Cockcroft and Gault) was significantly better at month 48 (M48) in the SRL group both in the intent-to-treat population (ITT): 62.6 mL/min/1.73 m(2) versus 57.1 mL/min/1.73 m(2) (p = 0.013) and in the on-treatment population (OT): 67.5 mL/min/1.73 m(2) versus 57.4 mL/min/1.73 m(2) (p = 0.002). Two biopsy proven acute rejection episodes occurred after M12 in each group. Graft and patient survival were comparable (graft survival: 97.4 vs. 100%; patient survival: 97.4 vs. 97.6%, respectively). The incidence of new-onset diabetes was numerically increased in the SRL group (7 vs. 2). In OT, three cancers occurred in the SRL group versus nine in the CsA group and mean proteinuria was increased in the SRL group (0.42 ± 0.44 vs. 0.26 ± 0.37; p = 0.018). In summary, the renal benefits associated with conversion of CsA to SRL, at 3 months posttransplantation, in combination with MMF were maintained for 4 years posttransplantation.

  5. User feedback on the MSF tele-expertise service after a four-year pilot trial –- a comprehensive analysis

    Directory of Open Access Journals (Sweden)

    Laurent eBONNARDOT

    2015-11-01

    Full Text Available We surveyed all users of the MSF tele-expertise service, approximately four years after it began operation. The survey contained 50 questions and was sent to 294 referrers and 254 specialists. There were 163 responses (response rate 30%. There were no significant differences between the responses from French and English users, so the responses were combined for subsequent analysis. Most of the responders were doctors (133 of 157 who answered that question, and most had completed field missions for MSF, i.e. both specialists and referrers. The majority stated that the system was user-friendly, and that they found it self-explanatory (i.e. they did not need to be shown how to use it. Almost all the referrers found that the telemedicine advice they received was helpful, and changed diagnosis, management and/or reassured the patient. Similar feedback came from the specialists, who also felt that there was educational value for the field doctor. Although there was general satisfaction with the service, the survey identified various problems. The main concerns -- raised by referrers was the lack of promotion of system at headquarters' level, and for specialists, the lack of feedback about patient follow up. Nonetheless, both referrers and specialists recognised the benefits of telemedicine in improving patient management, the provision of education and the reduction of isolation in the field.

  6. Searches for Extended and Point-like Neutrino Sources with Four Years of IceCube Data

    CERN Document Server

    Aartsen, M G; Adams, J; Aguilar, J A; Ahlers, M; Ahrens, M; Altmann, D; Anderson, T; Arguelles, C; Arlen, T C; Auffenberg, J; Bai, X; Barwick, S W; Baum, V; Beatty, J J; Tjus, J Becker; Becker, K -H; BenZvi, S; Berghaus, P; Berley, D; Bernardini, E; Bernhard, A; Besson, D Z; Binder, G; Bindig, D; Bissok, M; Blaufuss, E; Blumenthal, J; Boersma, D J; Bohm, C; Bos, F; Bose, D; Böser, S; Botner, O; Brayeur, L; Bretz, H -P; Brown, A M; Casey, J; Casier, M; Cheung, E; Chirkin, D; Christov, A; Christy, B; Clark, K; Classen, L; Clevermann, F; Coenders, S; Cowen, D F; Silva, A H Cruz; Danninger, M; Daughhetee, J; Davis, J C; Day, M; de André, J P A M; De Clercq, C; De Ridder, S; Desiati, P; de Vries, K D; de With, M; DeYoung, T; Díaz-Vélez, J C; Dunkman, M; Eagan, R; Eberhardt, B; Eichmann, B; Eisch, J; Euler, S; Evenson, P A; Fadiran, O; Fazely, A R; Fedynitch, A; Feintzeig, J; Felde, J; Feusels, T; Filimonov, K; Finley, C; Fischer-Wasels, T; Flis, S; Franckowiak, A; Frantzen, K; Fuchs, T; Gaisser, T K; Gallagher, J; Gerhardt, L; Gier, D; Gladstone, L; Glüsenkamp, T; Goldschmidt, A; Golup, G; Gonzalez, J G; Goodman, J A; Góra, D; Grandmont, D T; Grant, D; Gretskov, P; Groh, J C; Groß, A; Ha, C; Haack, C; Ismail, A Haj; Hallen, P; Hallgren, A; Halzen, F; Hanson, K; Hebecker, D; Heereman, D; Heinen, D; Helbing, K; Hellauer, R; Hellwig, D; Hickford, S; Hill, G C; Hoffman, K D; Hoffmann, R; Homeier, A; Hoshina, K; Huang, F; Huelsnitz, W; Hulth, P O; Hultqvist, K; Hussain, S; Ishihara, A; Jacobi, E; Jacobsen, J; Jagielski, K; Japaridze, G S; Jero, K; Jlelati, O; Jurkovic, M; Kaminsky, B; Kappes, A; Karg, T; Karle, A; Kauer, M; Kelley, J L; Kheirandish, A; Kiryluk, J; Kläs, J; Klein, S R; Köhne, J -H; Kohnen, G; Kolanoski, H; Koob, A; Köpke, L; Kopper, C; Kopper, S; Koskinen, D J; Kowalski, M; Kriesten, A; Krings, K; Kroll, G; Kroll, M; Kunnen, J; Kurahashi, N; Kuwabara, T; Labare, M; Larsen, D T; Larson, M J; Lesiak-Bzdak, M; Leuermann, M; Leute, J; Lünemann, J; Macías, O; Madsen, J; Maggi, G; Maruyama, R; Mase, K; Matis, H S; Maunu, R; McNally, F; Meagher, K; Medici, M; Meli, A; Meures, T; Miarecki, S; Middell, E; Middlemas, E; Milke, N; Miller, J; Mohrmann, L; Montaruli, T; Morse, R; Nahnhauer, R; Naumann, U; Niederhausen, H; Nowicki, S C; Nygren, D R; Obertacke, A; Odrowski, S; Olivas, A; Omairat, A; O'Murchadha, A; Palczewski, T; Paul, L; Penek, Ö; Pepper, J A; Heros, C Pérez de los; Pfendner, C; Pieloth, D; Pinat, E; Posselt, J; Price, P B; Przybylski, G T; Pütz, J; Quinnan, M; Rädel, L; Rameez, M; Rawlins, K; Redl, P; Rees, I; Reimann, R; Resconi, E; Rhode, W; Richman, M; Riedel, B; Robertson, S; Rodrigues, J P; Rongen, M; Rott, C; Ruhe, T; Ruzybayev, B; Ryckbosch, D; Saba, S M; Sander, H -G; Sandroos, J; Santander, M; Sarkar, S; Schatto, K; Scheriau, F; Schmidt, T; Schmitz, M; Schoenen, S; Schöneberg, S; Schönwald, A; Schukraft, A; Schulte, L; Schulz, O; Seckel, D; Sestayo, Y; Seunarine, S; Shanidze, R; Sheremata, C; Smith, M W E; Soldin, D; Spiczak, G M; Spiering, C; Stamatikos, M; Stanev, T; Stanisha, N A; Stasik, A; Stezelberger, T; Stokstad, R G; Stößl, A; Strahler, E A; Ström, R; Strotjohann, N L; Sullivan, G W; Taavola, H; Taboada, I; Tamburro, A; Tepe, A; Ter-Antonyan, S; Terliuk, A; Tešić, G; Tilav, S; Toale, P A; Tobin, M N; Tosi, D; Tselengidou, M; Unger, E; Usner, M; Vallecorsa, S; van Eijndhoven, N; Vandenbroucke, J; van Santen, J; Vehring, M; Voge, M; Vraeghe, M; Walck, C; Wallraff, M; Weaver, Ch; Wellons, M; Wendt, C; Westerhoff, S; Whelan, B J; Whitehorn, N; Wichary, C; Wiebe, K; Wiebusch, C H; Williams, D R; Wissing, H; Wolf, M; Wood, T R; Woschnagg, K; Xu, D L; Xu, X W; Yanez, J P; Yodh, G; Yoshida, S; Zarzhitsky, P; Ziemann, J; Zierke, S; Zoll, M

    2014-01-01

    We present results on searches for point-like sources of neutrinos using four years of IceCube data, including the first year of data from the completed 86-string detector. The total livetime of the combined dataset is 1,373 days. For an E$^{-2}$ spectrum the median sensitivity at 90\\% C.L. is $\\sim 10^{-12}$ TeV$^{-1}$cm$^{-2}$s$^{-1}$ for energies between 1 TeV$-$1 PeV in the northern sky and $\\sim 10^{-11}$ TeV$^{-1}$cm$^{-2}$s$^{-1}$ for energies between 100 TeV $-$ 100 PeV in the southern sky. The sensitivity has improved from both the additional year of data and the introduction of improved reconstructions compared to previous publications. In addition, we present the first results from an all-sky search for extended sources of neutrinos. We update results of searches for neutrino emission from stacked catalogs of sources, and test five new catalogs; two of Galactic supernova remnants and three of active galactic nuclei. In all cases, the data are compatible with the background-only hypothesis, and uppe...

  7. Integrating critical thinking and evidence-based dentistry across a four-year dental curriculum: a model for independent learning.

    Science.gov (United States)

    Marshall, Teresa A; Straub-Morarend, Cheryl L; Handoo, Nidhi; Solow, Catherine M; Cunningham-Ford, Marsha A; Finkelstein, Michael W

    2014-03-01

    Introducing critical thinking and evidence-based dentistry (EBD) content into an established dental curriculum can be a difficult and challenging process. Over the past three years, the University of Iowa College of Dentistry has developed and implemented a progressive four-year integrated critical thinking and EBD curriculum. The objective of this article is to describe the development and implementation process to make it available as a model for other dental schools contemplating introduction of critical thinking and EBD into their curricula. The newly designed curriculum built upon an existing problem-based learning foundation, which introduces critical thinking and the scientific literature in the D1 year, in order to expose students to the rationale and resources for practicing EBD in the D2 and D3 years and provide opportunities to practice critical thinking and apply the EBD five-step process in the D2, D3, and D4 years. All curricular content is online, and D3 and D4 EBD activities are integrated within existing clinical responsibilities. The curricular content, student resources, and student activities are described.

  8. Using time-on-task measurements to understand student performance in a physics class: A four-year study

    Directory of Open Access Journals (Sweden)

    John Stewart*

    2012-04-01

    Full Text Available Student use of out-of-class time was measured for four years in the introductory second-semester calculus-based physics course at the University of Arkansas. Two versions of the course were presented during the time of the measurement. In both versions, the total out-of-class time a student invested in the course explained less than 1% of the variance in test average and in normalized gain on the Conceptual Survey in Electricity and Magnetism. The details of how students used their out-of-class time explained from 21% to 36% of the variance in test average and 19% to 37% of the variance in normalized gain. The amount of time spent on the course increased as the course was made more difficult, but less than would be expected based on the changes in the course. Students changed their allocation of out-of-class time based on their performance within the course as the course progressed.

  9. Searches for extended and point-like neutrino sources with four years of IceCube data

    Energy Technology Data Exchange (ETDEWEB)

    Aartsen, M. G. [School of Chemistry and Physics, University of Adelaide, Adelaide SA, 5005 Australia (Australia); Ackermann, M.; Berghaus, P. [DESY, D-15735 Zeuthen (Germany); Adams, J. [Department of Physics and Astronomy, University of Canterbury, Private Bag 4800, Christchurch (New Zealand); Aguilar, J. A. [Département de physique Nucléaire et Corpusculaire, Université de Genève, CH-1211 Genève (Switzerland); Ahlers, M.; Arguelles, C.; BenZvi, S. [Department of Physics and Wisconsin IceCube Particle Astrophysics Center, University of Wisconsin, Madison, WI 53706 (United States); Ahrens, M. [Oskar Klein Centre and Department of Physics, Stockholm University, SE-10691 Stockholm (Sweden); Altmann, D. [Erlangen Centre for Astroparticle Physics, Friedrich-Alexander-Universität Erlangen-Nürnberg, D-91058 Erlangen (Germany); Anderson, T.; Arlen, T. C. [Department of Physics, Pennsylvania State University, University Park, PA 16802 (United States); Auffenberg, J. [III. Physikalisches Institut, RWTH Aachen University, D-52056 Aachen (Germany); Bai, X. [Physics Department, South Dakota School of Mines and Technology, Rapid City, SD 57701 (United States); Barwick, S. W. [Department of Physics and Astronomy, University of California, Irvine, CA 92697 (United States); Baum, V. [Institute of Physics, University of Mainz, Staudinger Weg 7, D-55099 Mainz (Germany); Beatty, J. J. [Department of Physics and Center for Cosmology and Astro-Particle Physics, Ohio State University, Columbus, OH 43210 (United States); Tjus, J. Becker [Fakultät für Physik and Astronomie, Ruhr-Universität Bochum, D-44780 Bochum (Germany); Becker, K.-H. [Department of Physics, University of Wuppertal, D-42119 Wuppertal (Germany); Berley, D. [Department of Physics, University of Maryland, College Park, MD 20742 (United States); Collaboration: IceCube Collaboration; and others

    2014-12-01

    We present results on searches for point-like sources of neutrinos using four years of IceCube data, including the first year of data from the completed 86 string detector. The total livetime of the combined data set is 1373 days. For an E {sup –2} spectrum, the observed 90% C.L. flux upper limits are ∼10{sup –12} TeV{sup –1} cm{sup –2} s{sup –1} for energies between 1 TeV and 1 PeV in the northern sky and ∼10{sup –11} TeV{sup –1} cm{sup –2} s{sup –1} for energies between 100 TeV and 100 PeV in the southern sky. This represents a 40% improvement compared to previous publications, resulting from both the additional year of data and the introduction of improved reconstructions. In addition, we present the first results from an all-sky search for extended sources of neutrinos. We update the results of searches for neutrino emission from stacked catalogs of sources and test five new catalogs; two of Galactic supernova remnants and three of active galactic nuclei. In all cases, the data are compatible with the background-only hypothesis, and upper limits on the flux of muon neutrinos are reported for the sources considered.

  10. Sex difference in response to stress by lunar month: A pilot study of four years' crisis-call frequency

    Directory of Open Access Journals (Sweden)

    Steffert Beverly

    2003-12-01

    Full Text Available Abstract Background This study is in response to the question of whether the moon can influence daily levels of stress. Method Four years of telephone-call frequency data were obtained from a single crisis-call centre. We used the method of lunar-day numbers 1 to 29 for analysis. We also tested the concept of 'strong moons' as occurring when the Sun was near to the lunar-node axis. Results An increase in calls was recorded from females and a decrease in calls by males during the new moon period, suggesting a sex difference in response, and there were proportionally more calls by males a fortnight later. A comparable swing in the male/female call-ratio on a weekly basis over Fridays and Saturdays was noted. Limitations of staffing at the call-centre prohibited comment on seasonal correlations. Conclusion Lunar-related studies of stress should endeavour to separate the data by sex or the effect can be lost. Distress-calls by women were more strongly linked to the lunar month than were those by men.

  11. Peripheral Giant Cell Granuloma in a Child Associated with Ectopic Eruption and Traumatic Habit with Control of Four Years

    Directory of Open Access Journals (Sweden)

    Luiz Evaristo Ricci Volpato

    2016-01-01

    Full Text Available Peripheral giant cell granuloma (PGCG is a nonneoplastic lesion that may affect any region of the gingiva or alveolar mucosa of edentulous and toothed areas, preferentially in the mandible and rarely occurring in children. This report describes the clinical and histopathological findings of a PGCG diagnosed in the maxilla of a 9-year-old boy associated with a tooth erupting improperly and a traumatic habit. The patient did not present anything noteworthy on extraoral physical examination or medical history, but the habit of picking his teeth and “poking” the gingiva. The oral lesion consisted of an asymptomatic, rounded, pink colored, smooth surface, soft tissue injury with fibrous consistency and approximated size of 1.5 cm located in the attached gingiva between the upper left permanent lateral incisor and the primary canine of the same side. Excisional biopsy was performed through curettage and removal of the periosteum, periodontal ligament, and curettage of the involved teeth with vestibular access. The histopathological analysis led to the diagnosis of PGCG. The prompt diagnosis and treatment of the PGCG resulted in a more conservative surgery and a reduced risk for tooth and bone loss and recurrence of the lesion. After four years of control, patient had no relapse of the lesion and good gingival and osseous health.

  12. Peripheral Giant Cell Granuloma in a Child Associated with Ectopic Eruption and Traumatic Habit with Control of Four Years

    Science.gov (United States)

    Leite, Cristhiane Almeida; Anhesini, Brunna Haddad; Aguilera, Jéssica Marques Gomes da Silva

    2016-01-01

    Peripheral giant cell granuloma (PGCG) is a nonneoplastic lesion that may affect any region of the gingiva or alveolar mucosa of edentulous and toothed areas, preferentially in the mandible and rarely occurring in children. This report describes the clinical and histopathological findings of a PGCG diagnosed in the maxilla of a 9-year-old boy associated with a tooth erupting improperly and a traumatic habit. The patient did not present anything noteworthy on extraoral physical examination or medical history, but the habit of picking his teeth and “poking” the gingiva. The oral lesion consisted of an asymptomatic, rounded, pink colored, smooth surface, soft tissue injury with fibrous consistency and approximated size of 1.5 cm located in the attached gingiva between the upper left permanent lateral incisor and the primary canine of the same side. Excisional biopsy was performed through curettage and removal of the periosteum, periodontal ligament, and curettage of the involved teeth with vestibular access. The histopathological analysis led to the diagnosis of PGCG. The prompt diagnosis and treatment of the PGCG resulted in a more conservative surgery and a reduced risk for tooth and bone loss and recurrence of the lesion. After four years of control, patient had no relapse of the lesion and good gingival and osseous health. PMID:27999690

  13. Lessons learned from four years of actively using River Forecast Center Ensemble Streamflow Predictions to inform reservoir management

    Science.gov (United States)

    Polebitski, A.; Palmer, R.; Meaker, B.

    2012-12-01

    The National Weather Service's River Forecast Centers (RFCs), located throughout the US, produce operational streamflow forecasts for short term application and long-term lead forecasts at selected locations. These forecasts are targeted for a variety of users, including water supply management, flood control, hydropower production, navigation, and recreation. This presentation highlights the challenges and successes associated with the use of RFC produced ensemble streamflow predictions (ESP) in generating system operations forecasts over the past four years for Snohomish County Public Utility District #1's (SnoPUD) Henry Jackson hydropower system. This research documents a multiyear collaboration between SnoPUD and academic researchers. The collaboration began with a proof of concept study in 2007 and evolved into a weekly decision support activity that has been ongoing since 2008 ( documented in Alemu et al. 2010). The Alemu et al. paper demonstrates the usefulness of ESP forecasts in hydropower operations decision making. This paper focuses on the value of forecasts and a decision support system (DSS) in improving skills in operating reservoir systems. During the application period, the model provided weekly guidance on meeting operational objectives and a probabilistic approach to quantifying system vulnerability during critical periods such as floods and drought. The ESP forecasts and the DSS were heavily used during periods of uncertainty and less so during periods of high system constraint or low system risk.

  14. Bone grafts in dentistry

    Directory of Open Access Journals (Sweden)

    Prasanna Kumar

    2013-01-01

    Full Text Available Bone grafts are used as a filler and scaffold to facilitate bone formation and promote wound healing. These grafts are bioresorbable and have no antigen-antibody reaction. These bone grafts act as a mineral reservoir which induces new bone formation.

  15. Chitosan-plasmid DNA nanoparticles encoding small hairpin RNA targeting MMP-3 and -13 to inhibit the expression of dedifferentiation related genes in expanded chondrocytes.

    Science.gov (United States)

    Zhao, Jingxin; Fan, Xiangli; Zhang, Qiang; Sun, Fangfei; Li, Xiaojian; Xiong, Chuan; Zhang, Chunli; Fan, Hongbin

    2014-02-01

    Overexpression of matrix metalloproteinase (MMP)-3 and -13 can lead to the dedifferentiation of expanded chondrocytes. After implanting dedifferentiated cells for cartilage defect repair, graft failure may occur. Short hairpin RNA (shRNA) is a powerful genetic tool to reduce the expression of target genes. This study investigated the effects of chitosan-plasmid DNA (pDNA) nanoparticles encoding shRNA targeting MMP-3 and -13 on the dedifferentiation of expanded chondrocytes. The objective was to optimize the parameters of chitosan-pDNA formulation for achieving higher efficiency of pDNA delivery and gene silencing. The chitosan-pDNA nanoparticles were prepared using a complex coacervation process. Then the characteristics including size, shape, stability, and transfection efficiency were compared in different groups. The results indicated that chitosan of 800 kDa at N/P ratio of 4 and pH 7.0 was optimal to prepare chitosan-pDNA nanoparticles. These nanoparticles showed high DNA loading efficiency (95.8 ± 1.5%) and high gene transfection efficiency (24.5 ± 1.6%). After the expanded chondrocytes were transfected by chitosan-pDNA nanoparticles, MMP-3-610 and MMP-13-2024 groups showed greater suppression in mRNA and protein levels. The results indicated that chitosan-pDNA nanoparticles encoding shRNA targeting MMP-3 and -13 had great potential in silencing the dedifferentiation-related genes for regenerating prolonged and endurable cartilage.

  16. Pilose antler polypeptides promote chondrocyte proliferation via the tyrosine kinase signaling pathway

    Directory of Open Access Journals (Sweden)

    Lin Jian-Hua

    2011-11-01

    Full Text Available Abstract Background Pilose antler polypeptides (PAP have been reported to promote chondrocyte proliferation. However, the underlying mechanism remains unclear. The present study was to investigate the effects of PAP on the proliferation of chondrocytes and its underlying mechanism. Methods Chondrocytes isolated from the knee of Zealand white rabbits were cultured. The second generation chondrocytes were collected and identified using safranin-O staining. The chondrocytes were divided into the following 4 groups including serum-free, PAP, genistein (an inhibitor of tyrosine kinases, and PAP plus genistein group. Cell viability was analyzed using the MTT assay. The cell cycle distribution of the chondrocytes was analyzed by flow cytometry. The expression levels of cyclin A was detected using immunocytochemical staining. Results No significant difference was observed between serum-free and genistein group. Treatment of the cultures with PAP produced a significant dose-dependent increase in cell viability, the percentage proportion of chondrocytes in the S phase and Cyclin A expression as well. However, the promoting effect of PAP on chondrocyte proliferation were dose-dependently inhibited by genistein, whereas genistein alone had no effect on proliferation of isolated chondrocytes. Conclusions The data demonstrate that PAP promotes chondrocyte proliferation with the increased cell number, percentage proportion of chondrocytes in S phase and expression of protein cyclin A via the TK signaling pathway.

  17. Growth characteristics and functional changes in rat chondrocytes cultured in porous tantalum in vitro

    Directory of Open Access Journals (Sweden)

    Ling ZHANG

    2014-08-01

    Full Text Available Objective To evaluate the growth characteristics and functional changes in rat chondrocytes cultured in porous tantalum in vitro. Methods The chondrocytes isolated from cartilage of 3-week old SD rats were cultured in vitro, then the 2nd passage cells were identified and implanted in porous tantalum scaffolds with a density of 1×106 cells/ml. The morphological characteristics of the chondrocytes cultured in porous tantalum were observed under inverted microscope, scanning electron microscope (SEM and transmission electron microscope (TEM, and the content of glycosaminoglycan (GAG in the chondrocytes was measured by chromatometry. Results The harvested cells were identified as chondrocytes by type Ⅱ collagen immunocytochemical staining, toluidine blue staining and safranin-O staining. Many chondrocytes adhering to the edge of porous tantalum were found by inverted microscope. Observation under SEM showed that chondrocytes spread well on the surface and distributed in the holes of porous tantalum, and they proliferated and secreted some extracellular matrixes. TEM observation showed that the ultrastructure of chondrocytes cultured in porous tantalum was similar to that of normal chondrocytes. Chromatometry determination showed that the chondrocytes in porous tantalum could secrete GAG continuously. Conclusion Porous tantalum is shown to have a satisfactory biocompatibility with chondrocytes in vitro, and may be used as a scaffold for cartilage tissue engineering. DOI: 10.11855/j.issn.0577-7402.2014.06.08

  18. Olecranon bone graft: revisited.

    Science.gov (United States)

    Mersa, Berkan; Ozcelik, Ismail Bulent; Kabakas, Fatih; Sacak, Bulent; Aydin, Atakan

    2010-09-01

    Autogenous bone grafts are frequently in use in the field of reconstructive upper extremity surgery. Cancellous bone grafts are applied to traumatic osseous defects, nonunions, defects after the resection of benign bone tumors, arthrodesis, and osteotomy procedures. Cancellous bone grafts do not only have benefits such as rapid revascularization, but they also have mechanical advantages. Despite the proximity to the primary surgical field, cancellous olecranon grafts have not gained the popularity they deserve in the field of reconstructive hand surgery. In this study, the properties, advantages, and technical details of harvesting cancellous olecranon grafts are discussed.

  19. Chondrogenic potential of articular chondrocytes depends on their original location

    NARCIS (Netherlands)

    Bekkers, Joris E J; Saris, Daniel B F; Tsuchida, Anika Iris; van Rijen, Mattie H P; Dhert, Wouter J A; Creemers, Laura B

    2014-01-01

    OBJECTIVE: This study aimed to investigate the regenerative capacity of chondrocytes derived from debrided defect cartilage and healthy cartilage from different regions in the joint to determine the best cell source for regenerative cartilage therapies. METHODS: Articular cartilage was obtained from

  20. MicroRNA-33 suppresses CCL2 expression in chondrocytes.

    Science.gov (United States)

    Wei, Meng; Xie, Qingyun; Zhu, Jun; Wang, Tao; Zhang, Fan; Cheng, Yue; Guo, Dongyang; Wang, Ying; Mo, Liweng; Wang, Shuai

    2016-06-01

    CCL2-mediated macrophage infiltration in articular tissues plays a pivotal role in the development of the osteoarthritis (OA). miRNAs regulate the onset and progression of diseases via controlling the expression of a series of genes. How the CCL2 gene was regulated by miRNAs was still not fully elucidated. In the present study, we demonstrated that the binding sites of miR-33 in the 3'UTR of CCL2 gene were conserved in human, mouse and rat species. By performing gain- or loss-of-function studies, we verified that miR-33 suppressed CCL2 expression in the mRNA and protein levels. We also found that miR-33 suppressed the CCL2 levels in the supernatant of cultured primary mouse chondrocytes. With reporter gene assay, we demonstrated that miR-33 targeted at AAUGCA in the 3'UTR of CCL2 gene. In transwell migration assays, we demonstrated that the conditional medium (CM) from miR-33 deficient chondrocytes potentiated the monocyte chemotaxis in a CCL2 dependent manner. Finally, we demonstrated that the level of miR-33 was decreased, whereas the CCL2 level was increased in the articular cartilage from the OA patients compared with the control group. In summary, we identified miR-33 as a novel suppressor of CCL2 in chondrocytes. The miR-33/CCL2 axis in chondrocytes regulates monocyte chemotaxis, providing a potential mechanism of macrophage infiltration in OA.

  1. Confocal microscopy indentation system for studying in situ chondrocyte mechanics.

    Science.gov (United States)

    Han, Sang-Kuy; Colarusso, Pina; Herzog, Walter

    2009-10-01

    Chondrocytes synthesize extracellular matrix molecules, thus they are essential for the development, adaptation and maintenance of articular cartilage. Furthermore, it is well accepted that the biosynthetic activity of chondrocytes is influenced by the mechanical environment. Therefore, their response to mechanical stimuli has been studied extensively. Much of the knowledge in this area of research has been derived from testing of isolated cells, cartilage explants, and fixed cartilage specimens: systems that differ in important aspects from chondrocytes embedded in articular cartilage and observed during loading conditions. In this study, current model systems have been improved by working with the intact cartilage in real time. An indentation system was designed on a confocal microscope that allows for simultaneous loading and observation of chondrocytes in their native environment. Cell mechanics were then measured under precisely controlled loading conditions. The indentation system is based on a light transmissible cylindrical glass indentor of 0.17 mm thickness and 1.64 mm diameter that is aligned along the focal axis of the microscope and allows for real time observation of live cells in their native environment. The system can be used to study cell deformation and biological responses, such as calcium sparks, while applying prescribed loads on the cartilage surface. It can also provide novel information on the relationship between cell loading and cartilage adaptive/degenerative processes in the intact tissue.

  2. Cartilage homeoprotein 1, a homeoprotein selectively expressed in chondrocytes.

    Science.gov (United States)

    Zhao, G Q; Zhou, X; Eberspaecher, H; Solursh, M; de Crombrugghe, B

    1993-09-15

    We identified a rat cDNA that encodes cartilage homeoprotein 1 (Cart-1). The deduced amino acid sequence of Cart-1 contains a paired-type homeodomain. Northern blot hybridization and RNase protection assay revealed that Cart-1 RNA was present at high levels in a well-differentiated rat chondrosarcoma tumor and in a cell line derived from this tumor. Cart-1 RNA was detected in primary mouse and rat chondrocytes but not in various fibroblasts including mouse 10T1/2 cells, NIH 3T3 cells, BALB 3T3 cells, and rat skin fibroblasts. It was also undetectable in mouse C2 myoblasts, S194 myeloma cells, and embryonic stem cells. Cart-1 RNA was present at a very low level in tested but was not detected in other soft tissues of 8-week-old rats. In situ hybridization of rat embryos between 14.5 and 16.5 days post coitum revealed relatively high levels of Cart-1 RNA in condensed prechondrocytic mesenchymal cells and in early chondrocytes of cartilage primordia. The levels of Cart-1 RNA were lower in mature chondrocytes. No hybridization was observed in brain, spinal cord, heart, spleen, gastrointestinal tract, liver, and muscle. We speculate that Cart-1 has a role in chondrocyte differentiation.

  3. In vitro exposure of human chondrocytes to pulsed electromagnetic fields

    Directory of Open Access Journals (Sweden)

    V Nicolin

    2009-08-01

    Full Text Available The effect of pulsed electromagnetic fields (PEMFs on the proliferation and survival of matrix-induced autologous chondrocyte implantation (MACI®-derived cells was studied to ascertain the healing potential of PEMFs. MACI-derived cells were taken from cartilage biopsies 6 months after surgery and cultured. No dedifferentiation towards the fibroblastic phenotype occurred, indicating the success of the surgical implantation. The MACI-derived cultured chondrocytes were exposed to 12 h/day (short term or 4 h/day (long term PEMFs exposure (magnetic field intensity, 2 mT; frequency, 75 Hz and proliferation rate determined by flow cytometric analysis. The PEMFs exposure elicited a significant increase of cell number in the SG2M cell cycle phase. Moreover, cells isolated from MACI® scaffolds showed the presence of collagen type II, a typical marker of chondrocyte functionality. The results show that MACI® membranes represent an optimal bioengineering device to support chondrocyte growth and proliferation in surgical implants. The surgical implant of MACI® combined with physiotherapy is suggested as a promising approach for a faster and safer treatment of cartilage traumatic lesions.

  4. Doxycycline inhibits collagen synthesis by differentiated articular chondrocytes.

    NARCIS (Netherlands)

    TeKoppele, J.M.; Beekman, B.; Verzijl, N.; Koopman, J.L.; Groot, J. de; Bank, R.A.

    1998-01-01

    Doxycycline (DOX) profoundly inhibited collagen synthesis by differentiated articular chondrocytes. At 25 microM, the rate of collagen synthesis was suppressed by more than 50% without affecting cell proliferation (DNA levels) and general protein synthesis (35S-Met and 35S-Cys incorporation). Steady

  5. Experiences of internet-delivered cognitive behavior therapy for social anxiety disorder four years later: A qualitative study

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    Camilla Olsson Halmetoja

    2014-07-01

    Full Text Available The current study is a qualitative follow-up of a study on guided internet-delivered cognitive behavior therapy (ICBT for social anxiety disorder (SAD, conducted four years after treatment completion. The main aim was to capture participants' description of their experiences of the treatment, their view on treatment effects, memories of the treatment, and whether they continued using the gained knowledge after treatment. Sixty participants were selected from the original study's treatment group. A criterion based sampling approach was used based on the obtained treatment effect, and with a minimum of five completed treatment modules. E-mail invitations were sent, with information about the follow-up and the instruction to respond if interested in participating. Twelve semi-structured interviews were made and the material was analyzed using an approach based on grounded theory. The results showed that all participants found the treatment to have some effect, but they also found it to be demanding, difficult, and hard. Many appreciated to hear of the experiences of other participants in the online forum. Under the theme of memory, most could describe the setup of the treatment in general terms. The exposure module was mentioned by all, cognitive restructuring by most, and some also reported memories of the psychoeducation. A core process was identified which involved how the attained treatment effect was viewed over the time, and how this view changed from treatment completion to current time. The findings outlined in this study describe how treatment effects can be sustained via an active approach to the treatment and the symptoms of SAD.

  6. Port Pirie Cohort Study: environmental exposure to lead and children's abilities at the age of four years

    Energy Technology Data Exchange (ETDEWEB)

    McMichael, A.J.; Baghurst, P.A.; Wigg, N.R.; Vimpani, G.V.; Robertson, E.F.; Roberts, R.J.

    1988-08-25

    We studied the effect of environmental exposure to lead on children's abilities at the age of four years in a cohort of 537 children born during 1979 to 1982 to women living in a community situated near a lead smelter. Samples for measuring blood lead levels were obtained from the mothers antenatally, at delivery from the mothers and umbilical cords, and at the ages of 6, 15, and 24 months and then annually from the children. Concurrently, the mothers were interviewed about personal, family, medical, and environmental factors. Maternal intelligence, the home environment, and the children's mental development (as evaluated with use of the McCarthy Scales of Children's Abilities) were formally assessed. The mean blood lead concentration varied from 0.44 mumol per liter in midpregnancy to a peak of 1.03 mumol per liter at the age of two years. The blood lead concentration at each age, particularly at two and three years, and the integrated postnatal average concentration were inversely related to development at the age of four. Multivariate analysis incorporating many factors in the children's lives indicated that the subjects with an average postnatal blood lead concentration of 1.50 mumol per liter had a general cognitive score 7.2 points lower (95 percent confidence interval, 0.3 to 13.2; mean score, 107.1) than those with an average concentration of 0.50 mumol per liter. Similar deficits occurred in the perceptual-performance and memory scores. Within the range of exposure studied, no threshold dose for an effect of lead was evident. We conclude that postnatal blood lead concentration is inversely related to cognitive development in children, although one must be circumspect in making causal inferences from studies of this relation, because of the difficulties in defining and controlling confounding effects.

  7. Donor-specific HLA antibodies in chronic renal allograft rejection: a prospective trial with a four-year follow-up.

    Science.gov (United States)

    Lachmann, Nils; Terasaki, Paul I; Schönemann, Constanze

    2006-01-01

    A total of 1,043 serum samples of kidney allograft recipients with functioning grafts at a mean of 6 years after transplantation were tested once for HLA abs and monitored for graft and patient survival for 4 years after testing. All grafts were transplanted between 1984 and 2004 at the Charité hospital (Berlin, Germany). 1. To our knowledge this is the first study with more than 1,000 patients designed to elucidate the impact of DSA and NDSA on late renal graft outcome. True donor specificity of HLA abs in 30% of antibody positive patients was determined by using a single HLA antigen coated bead assay. 2. The long-term graft survival was adversely impacted by the presence of HLA abs directed against mismatched HLA antigens and to a lesser extent also by NDSA. This difference was shown to be leveled at late stages of graft damage, suggesting NDSA as a predictor of adsorbed DSA killing the graft. 3. The deleterious effect of preexisting and de novo formed HLA abs on long-term graft function was shown to be equal. Despite a negative crossmatch at the time of transplantation sensitized recipients and those who produced HLA abs de novo posttransplant showed the same increased risk of late graft failure. 4. Among patients with DSA and additional NDSA the majority of 74% of assigned potential HLA class I epitopes were shared among DSA and NDSA suggesting mismatched donor antigens as the cause of detected NDSA.

  8. Bacterial urinary tract infection in renal transplant recipients and their antibiotic resistance pattern: A four-year study.

    Directory of Open Access Journals (Sweden)

    Azar Dokht Khosravi

    2014-04-01

    Full Text Available Urinary tract infections (UTIs are the most common infections in renal transplant recipients and are considered a potential cause of bacteremia, sepsis, and affects graft outcomes. The aim of the present study was to determine the incidence of UTI among renal transplant recipients and investigation of antimicrobial susceptibility pattern of causative agents.In total, 1165 patients from March 2009 to December 2012, in transplant center of Golestan Hospital, Ahvaz, Iran, were investigated. Qualitative urine cultures were performed for all cases, causative microorganisms were identified and colony count was performed according to the standard protocol. Antibiotic susceptibility testing was then performed to determine the susceptibility pattern of recovered bacteria from confirmed UTIs.UTI was diagnosed in 391 patients(33.56%. Gram-negative bacteria were the most prevalent isolated microorganisms with E. coli (43.53%, followed by Enterobacter spp. (35.37% as the major organisms. Among Gram positives, Coagulase-negative Staphylococci was isolated from 6.8% of cases. The rate of resistance to all tested antibiotics was highest in Enterobacter spp., however the most common resistance were seen against cefixime, cephalotin, and cotrimoxazole in all tested gram negatives.the rate of UTIs among renal transplant recipients was noticeable in this study with high antibiotic resistance. Multi-resistant bacterial infections are potentially life-threatening emerging problem in renal transplantation. Prophylactic measures must be applied to patients at greater risk.

  9. A global survey of aerosol-liquid water cloud overlap based on four years of CALIPSO-CALIOP data

    Directory of Open Access Journals (Sweden)

    A. Devasthale

    2010-09-01

    Full Text Available The presence of aerosols over highly reflective liquid water cloud tops poses a big challenge in simulating their radiative impacts. Particularly, absorbing aerosols, such as smoke, may have significant impact in such situations and even change the sign of net radiative forcing. Until now, it was not possible to obtain information on such overlap events realistically from the existing passive satellite sensors. However, the CALIOP instrument onboard NASA's CALIPSO satellite allows us to examine these events with an unprecedented accuracy.

    Using four years of collocated CALIPSO 5 km Aerosol and Cloud Layer Version 3 Products (June 2006–May 2010, we quantify, for the first time, the macrophysical characteristics of overlapping aerosol and water cloud layers globally. We investigate seasonal variability in these characteristics over six latitude bands to understand the hemispheric differences. We compute a the percentage cases when such overlap is seen globally and seasonally when all aerosol types are included (AAO case in the analysis, b the joint histograms of aerosol layer base height and cloud layer top height, and c the joint histograms of aerosol and cloud geometrical thicknesses in such overlap cases. We also investigate frequency of smoke aerosol-cloud overlap (SAO case.

    The results show a distinct seasonality in overlap frequency in both AAO and SAO cases. Globally, the frequency is highest during JJA months in AAO case, while for the SAO case, it is highest in SON months. The seasonal mean overlap frequency can regionally exceed 20% in AAO case and 10% in SAO case. There is a tendency that the vertical separation between aerosol and cloud layers increases from high to low latitude regions in the both hemispheres. In about 5–10% cases the vertical distance between aerosol and cloud layers is less than 100 m, while about in 45–60% cases it less than a kilometer in the annual means for different latitudinal bands

  10. PKS 2005-489 at VHE: four years of monitoring with HESS and simultaneous multi-wavelength observations

    Science.gov (United States)

    H.E.S.S. Collaboration; Acero, F.; Aharonian, F.; Akhperjanian, A. G.; Anton, G.; Barres de Almeida, U.; Bazer-Bachi, A. R.; Becherini, Y.; Behera, B.; Benbow, W.; Bernlöhr, K.; Bochow, A.; Boisson, C.; Bolmont, J.; Borrel, V.; Brucker, J.; Brun, F.; Brun, P.; Bühler, R.; Bulik, T.; Büsching, I.; Boutelier, T.; Chadwick, P. M.; Charbonnier, A.; Chaves, R. C. G.; Cheesebrough, A.; Chounet, L.-M.; Clapson, A. C.; Coignet, G.; Costamante, L.; Dalton, M.; Daniel, M. K.; Davids, I. D.; Degrange, B.; Deil, C.; Dickinson, H. J.; Djannati-Ataï, A.; Domainko, W.; O'C. Drury, L.; Dubois, F.; Dubus, G.; Dyks, J.; Dyrda, M.; Egberts, K.; Eger, P.; Espigat, P.; Fallon, L.; Farnier, C.; Fegan, S.; Feinstein, F.; Fiasson, A.; Förster, A.; Fontaine, G.; Füßling, M.; Gabici, S.; Gallant, Y. A.; Gérard, L.; Gerbig, D.; Giebels, B.; Glicenstein, J. F.; Glück, B.; Goret, P.; Göring, D.; Hauser, M.; Heinz, S.; Heinzelmann, G.; Henri, G.; Hermann, G.; Hinton, J. A.; Hoffmann, A.; Hofmann, W.; Hofverberg, P.; Holleran, M.; Hoppe, S.; Horns, D.; Jacholkowska, A.; de Jager, O. C.; Jahn, C.; Jung, I.; Katarzyński, K.; Katz, U.; Kaufmann, S.; Kerschhaggl, M.; Khangulyan, D.; Khélifi, B.; Keogh, D.; Klochkov, D.; Kluźniak, W.; Kneiske, T.; Komin, Nu.; Kosack, K.; Kossakowski, R.; Lamanna, G.; Lenain, J.-P.; Lohse, T.; Marandon, V.; Martineau-Huynh, O.; Marcowith, A.; Masbou, J.; Maurin, D.; McComb, T. J. L.; Medina, M. C.; Méhault, J.; Moderski, R.; Moulin, E.; Naumann-Godo, M.; de Naurois, M.; Nedbal, D.; Nekrassov, D.; Nicholas, B.; Niemiec, J.; Nolan, S. J.; Ohm, S.; Olive, J.-F.; de Oña Wilhelmi, E.; Orford, K. J.; Ostrowski, M.; Panter, M.; Paz Arribas, M.; Pedaletti, G.; Pelletier, G.; Petrucci, P.-O.; Pita, S.; Pühlhofer, G.; Punch, M.; Quirrenbach, A.; Raubenheimer, B. C.; Raue, M.; Rayner, S. M.; Renaud, M.; Rieger, F.; Ripken, J.; Rob, L.; Rosier-Lees, S.; Rowell, G.; Rudak, B.; Rulten, C. B.; Ruppel, J.; Sahakian, V.; Santangelo, A.; Schlickeiser, R.; Schöck, F. M.; Schwanke, U.; Schwarzburg, S.; Schwemmer, S.; Shalchi, A.; Sikora, M.; Skilton, J. L.; Sol, H.; Stawarz, Ł.; Steenkamp, R.; Stegmann, C.; Stinzing, F.; Superina, G.; Szostek, A.; Tam, P. H.; Tavernet, J.-P.; Terrier, R.; Tibolla, O.; Tluczykont, M.; van Eldik, C.; Vasileiadis, G.; Venter, C.; Venter, L.; Vialle, J. P.; Vincent, P.; Vivier, M.; Völk, H. J.; Volpe, F.; Wagner, S. J.; Ward, M.; Zdziarski, A. A.; Zech, A.

    2010-02-01

    Aims: Our aim is to study the very high energy (VHE; E>100 GeV) γ-ray emission from BL Lac objects and the evolution in time of their broad-band spectral energy distribution (SED). Methods: VHE observations of the high-frequency peaked BL Lac object PKS 2005-489 were made with the High Energy Stereoscopic System (HESS) from 2004 through 2007. Three simultaneous multi-wavelength campaigns at lower energies were performed during the HESS data taking, consisting of several individual pointings with the XMM-Newton and RXTE satellites. Results: A strong VHE signal, ~17σ total, from PKS 2005-489 was detected during the four years of HESS observations (90.3 h live time). The integral flux above the average analysis threshold of 400 GeV is ~3% of the flux observed from the Crab Nebula and varies weakly on time scales from days to years. The average VHE spectrum measured from ~300 GeV to ~5 TeV is characterized by a power law with a photon index, Γ = 3.20± 0.16_stat± 0.10_syst. At X-ray energies the flux is observed to vary by more than an order of magnitude between 2004 and 2005. Strong changes in the X-ray spectrum (ΔΓX ≈ 0.7) are also observed, which appear to be mirrored in the VHE band. Conclusions: The SED of PKS 2005-489, constructed for the first time with contemporaneous data on both humps, shows significant evolution. The large flux variations in the X-ray band, coupled with weak or no variations in the VHE band and a similar spectral behavior, suggest the emergence of a new, separate, harder emission component in September 2005. Supported by CAPES Foundation, Ministry of Education of Brazil.Now at Harvard-Smithsonian Center for Astrophysics, Cambridge, USA.Now at W.W. Hansen Experimental Physics Laboratory & Kavli Institute for Particle Astrophysics and Cosmology, Stanford University, Stanford, USA.

  11. A global survey of aerosol-liquid water cloud overlap based on four years of CALIPSO-CALIOP data

    Science.gov (United States)

    Devasthale, A.; Thomas, M. A.

    2011-02-01

    Simulating the radiative impacts of aerosols located above liquid water clouds presents a significant challenge. In particular, absorbing aerosols, such as smoke, may have significant impact in such situations and even change the sign of net radiative forcing. It is not possible to reliably obtain information on such overlap events from existing passive satellite sensors. However, the CALIOP instrument onboard NASA's CALIPSO satellite allows us to examine these events with unprecedented accuracy. Using four years of collocated CALIPSO 5 km Aerosol and Cloud Layer Version 3 Products (June 2006-May 2010), we quantify, for the first time, the characteristics of overlapping aerosol and water cloud layers globally. We investigate seasonal variability in these characteristics over six latitude bands to understand the hemispheric differences when all aerosol types are included in the analysis (the AAO case). We also investigate frequency of smoke aerosol-cloud overlap (the SAO case). Globally, the frequency is highest during the JJA months in the AAO case, while for the SAO case, it is highest in the SON months. The seasonal mean overlap frequency can regionally exceed 20% in the AAO case and 10% in the SAO case. In about 5-10% cases the vertical distance between aerosol and cloud layers is less than 100 m, while about in 45-60% cases it less than a kilometer in the annual means for different latitudinal bands. In about 70-80% cases, aerosol layers are less than a kilometer thick, while in about 18-22% cases they are 1-2 km thick. The frequency of aerosol layers 2-3 km thick is about 4-5% in the tropical belts during overlap events. Over the regions where high aerosol loadings are present, the overlap frequency can be up to 50% higher when quality criteria on aerosol/cloud feature detection are relaxed. Over the polar regions, more than 50% of the overlapping aerosol layers have optical thickness less than 0.02, but the contribution from the relatively optically thicker

  12. A global survey of aerosol-liquid water cloud overlap based on four years of CALIPSO-CALIOP data

    Directory of Open Access Journals (Sweden)

    A. Devasthale

    2011-02-01

    Full Text Available Simulating the radiative impacts of aerosols located above liquid water clouds presents a significant challenge. In particular, absorbing aerosols, such as smoke, may have significant impact in such situations and even change the sign of net radiative forcing. It is not possible to reliably obtain information on such overlap events from existing passive satellite sensors. However, the CALIOP instrument onboard NASA's CALIPSO satellite allows us to examine these events with unprecedented accuracy. Using four years of collocated CALIPSO 5 km Aerosol and Cloud Layer Version 3 Products (June 2006–May 2010, we quantify, for the first time, the characteristics of overlapping aerosol and water cloud layers globally. We investigate seasonal variability in these characteristics over six latitude bands to understand the hemispheric differences when all aerosol types are included in the analysis (the AAO case. We also investigate frequency of smoke aerosol-cloud overlap (the SAO case. Globally, the frequency is highest during the JJA months in the AAO case, while for the SAO case, it is highest in the SON months. The seasonal mean overlap frequency can regionally exceed 20% in the AAO case and 10% in the SAO case. In about 5–10% cases the vertical distance between aerosol and cloud layers is less than 100 m, while about in 45–60% cases it less than a kilometer in the annual means for different latitudinal bands. In about 70–80% cases, aerosol layers are less than a kilometer thick, while in about 18–22% cases they are 1–2 km thick. The frequency of aerosol layers 2–3 km thick is about 4–5% in the tropical belts during overlap events. Over the regions where high aerosol loadings are present, the overlap frequency can be up to 50% higher when quality criteria on aerosol/cloud feature detection are relaxed. Over the polar regions, more than 50% of the overlapping aerosol layers have optical thickness less than 0.02, but the contribution from

  13. Spatiotemporal Determinants of Urban Leptospirosis Transmission: Four-Year Prospective Cohort Study of Slum Residents in Brazil.

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    José E Hagan

    2016-01-01

    Full Text Available Rat-borne leptospirosis is an emerging zoonotic disease in urban slum settlements for which there are no adequate control measures. The challenge in elucidating risk factors and informing approaches for prevention is the complex and heterogeneous environment within slums, which vary at fine spatial scales and influence transmission of the bacterial agent.We performed a prospective study of 2,003 slum residents in the city of Salvador, Brazil during a four-year period (2003-2007 and used a spatiotemporal modelling approach to delineate the dynamics of leptospiral transmission. Household interviews and Geographical Information System surveys were performed annually to evaluate risk exposures and environmental transmission sources. We completed annual serosurveys to ascertain leptospiral infection based on serological evidence. Among the 1,730 (86% individuals who completed at least one year of follow-up, the infection rate was 35.4 (95% CI, 30.7-40.6 per 1,000 annual follow-up events. Male gender, illiteracy, and age were independently associated with infection risk. Environmental risk factors included rat infestation (OR 1.46, 95% CI, 1.00-2.16, contact with mud (OR 1.57, 95% CI 1.17-2.17 and lower household elevation (OR 0.92 per 10m increase in elevation, 95% CI 0.82-1.04. The spatial distribution of infection risk was highly heterogeneous and varied across small scales. Fixed effects in the spatiotemporal model accounted for the majority of the spatial variation in risk, but there was a significant residual component that was best explained by the spatial random effect. Although infection risk varied between years, the spatial distribution of risk associated with fixed and random effects did not vary temporally. Specific "hot-spots" consistently had higher transmission risk during study years.The risk for leptospiral infection in urban slums is determined in large part by structural features, both social and environmental. Our findings

  14. Conditional expression of constitutively active estrogen receptor {alpha} in chondrocytes impairs longitudinal bone growth in mice

    Energy Technology Data Exchange (ETDEWEB)

    Ikeda, Kazuhiro [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Tsukui, Tohru [Experimental Animal Laboratory, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Imazawa, Yukiko; Horie-Inoue, Kuniko [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Inoue, Satoshi, E-mail: INOUE-GER@h.u-tokyo.ac.jp [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Department of Geriatric Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo (Japan); Department of Anti-Aging Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo (Japan)

    2012-09-07

    Highlights: Black-Right-Pointing-Pointer Conditional transgenic mice expressing constitutively active estrogen receptor {alpha} (caER{alpha}) in chondrocytes were developed. Black-Right-Pointing-Pointer Expression of caER{alpha} in chondrocytes impaired longitudinal bone growth in mice. Black-Right-Pointing-Pointer caER{alpha} affects chondrocyte proliferation and differentiation. Black-Right-Pointing-Pointer This mouse model is useful for understanding the physiological role of ER{alpha}in vivo. -- Abstract: Estrogen plays important roles in the regulation of chondrocyte proliferation and differentiation, which are essential steps for longitudinal bone growth; however, the mechanisms of estrogen action on chondrocytes have not been fully elucidated. In the present study, we generated conditional transgenic mice, designated as caER{alpha}{sup ColII}, expressing constitutively active mutant estrogen receptor (ER) {alpha} in chondrocytes, using the chondrocyte-specific type II collagen promoter-driven Cre transgenic mice. caER{alpha}{sup ColII} mice showed retardation in longitudinal growth, with short bone lengths. BrdU labeling showed reduced proliferation of hypertrophic chondrocytes in the proliferating layer of the growth plate of tibia in caER{alpha}{sup ColII} mice. In situ hybridization analysis of type X collagen revealed that the maturation of hypertrophic chondrocytes was impaired in caER{alpha}{sup ColII} mice. These results suggest that ER{alpha} is a critical regulator of chondrocyte proliferation and maturation during skeletal development, mediating longitudinal bone growth in vivo.

  15. Changes in particulate matter physical properties during Saharan advections over Rome (Italy: a four-year study, 2001–2004

    Directory of Open Access Journals (Sweden)

    R. Sozzi

    2013-02-01

    Full Text Available Particulate matter mass concentrations measured in the city of Rome (Italy in the period 2001–2004 have been cross-analysed with concurrent Saharan dust advection events to infer the impact these natural episodes bear on the standard air quality parameter PM10 observed at two city stations and at one regional background station. Natural events as Saharan dust advections are associated to a definite health risk. At the same time, the Directive 2008/50/EC allows subtraction of PM exceedances caused by natural contributions from statistics used to determine air-quality of EU sites. In this respect, it is important to detect and characterize such advections by means of reliable, operational techniques. To assess the PM10 increase we used both the "regional-background method" suggested by EC Guidelines and a "local background" one, demonstrated to be most suited to this central Mediterranean region. The two approaches provided results within 20% from each other. The sequence of Saharan advections over the city has been either detected by Polarization Lidar (laser radar observations or forecast by the operational numerical regional mineral dust model BSC-DREAM8b of the Barcelona Supercomputing Centre. Lidar observations were also employed to retrieve the average physical properties of the dust clouds as a function of height. Along the four-year period, Lidar measurements (703 evenly distributed days revealed Saharan plumes transits over Rome on 28.6% of the days, with minimum occurrence in wintertime. Dust was observed to reach the ground on 17.5% of the days totalling 88 episodes. Most (90% of these advections lasted up to 5 days, averaging to ~3 days. Median time lag between advections was 7 days. Typical altitude range of the dust plumes was 0–6 km, with centre of mass at ~3 km a.g.l. BSC-DREAM8b model simulations (1461 days predicted Lidar detectable (532nm extinction coefficient >0.005 km−1 dust advections on 25.9% of the days, with ground

  16. Four-year measurement of methane flux over a temperate forest with a relaxed eddy accumulation method

    Science.gov (United States)

    Sakabe, A.; Kosugi, Y.; Ueyama, M.; Hamotani, K.; Takahashi, K.; Iwata, H.; Itoh, M.

    2013-12-01

    Forests are generally assumed to be an atmospheric methane (CH4) sink (Le Mer and Roger, 2001). However, under Asian monsoon climate, forests are subject to wide spatiotemporal range in soil water status, where forest soils often became water-saturated condition heterogeneously. In such warm and humid conditions, forests may act as a CH4 source and/or sink with considerable spatiotemporal variations. Micrometeorological methods such as eddy covariance (EC) method continuously measure spatially-representative flux at a canopy scale without artificial disturbance. In this study, we measured CH4 fluxes over a temperate forest during four-year period using a CH4 analyzer based on tunable diode laser spectroscopy detection with a relaxed eddy accumulation (REA) method (Hamotani et al., 1996, 2001). We revealed the amplitude and seasonal variations of canopy-scale CH4 fluxes. The REA method is the attractive alternative to the EC method to measure trace-gas flux because it allows the use of analyzers with an optimal integration time. We also conducted continuous chamber measurements on forest floor to reveal spatial variations in soil CH4 fluxes and its controlling processes. The observations were made in an evergreen coniferous forest in central Japan. The site has a warm temperate monsoon climate with wet summer. Some wetlands were located in riparian zones along streams within the flux footprint area. For the REA method, the sonic anemometer (SAT-550, Kaijo) was mounted on top of the 29-m-tall tower and air was sampled from just below the sonic anemometer to reservoirs according to the direction of vertical wind velocity (w). After accumulating air for 30 minutes, the air in the reservoirs was pulled into a CO2/H2O gas analyzer (LI-840, Li-Cor) and a CH4 analyzer (FMA-200, Los Gatos Research). Before entering the analyzers, the sampled air was dried using a gas dryer (PD-50 T-48; Perma Pure Inc.). The REA flux is obtained from the difference in the mean concentrations

  17. Access to Chondrocyte Culture, with Alginate, In Iran

    Directory of Open Access Journals (Sweden)

    Ebrahim Esfandiary

    2008-01-01

    Full Text Available In this study, chondrocyte culture was established for the first time in Iran,and calcium alginate was used for longer culture of chondrocyte in vitro. Thestudy was programmed in order to be used for future human chondrocytetransplantation. The cartilage specimen obtained from 50 patients whounderwent total knee and hip operations in Isfahan University of MedicalSciences. Cartilage specimens were used for monolayer as well as suspensionculture in alginate beads. Approximately 12±1 millions cells were harvestedfrom the 3rd passage. The cells were round with large euchromatic nucleusand several nucleoli and small vacuoles. The cells derived from passages 1to 4, which were grown up then, in alginate beads, showed higher stainingwith alcian blue. The harvested cells in some patients were immediately andsuccessfully used for autologus transplantation. This later work will be reportedseparately.

  18. Interplay between cytoskeletal polymerization and the chondrogenic phenotype in chondrocytes passaged in monolayer culture.

    Science.gov (United States)

    Parreno, Justin; Nabavi Niaki, Mortah; Andrejevic, Katarina; Jiang, Amy; Wu, Po-Han; Kandel, Rita A

    2017-02-01

    Tubulin and actin exist as monomeric units that polymerize to form either microtubules or filamentous actin. As the polymerization status (monomeric/polymeric ratio) of tubulin and/or actin have been shown to be important in regulating gene expression and phenotype in non-chondrocyte cells, the objective of this study was to examine the role of cytoskeletal polymerization on the chondrocyte phenotype. We hypothesized that actin and/or tubulin polymerization status modulates the chondrocyte phenotype during monolayer culture as well as in 3D culture during redifferentiation. To test this hypothesis, articular chondrocytes were grown and passaged in 2D monolayer culture. Cell phenotype was investigated by assessing cell morphology (area and circularity), actin/tubulin content, organization and polymerization status, as well as by determination of proliferation, fibroblast and cartilage matrix gene expression with passage number. Bovine chondrocytes became larger, more elongated, and had significantly (P  0.05) modulated, actin polymerization was increased in bovine P2 cells. Actin depolymerization, but not tubulin depolymerization, promoted the chondrocyte phenotype by inducing cell rounding, increasing aggrecan and reducing COL1 expression. Knockdown of actin depolymerization factor, cofilin, in these cells induced further P2 cell actin polymerization and increased COL1 gene expression. To confirm that actin status regulated COL1 gene expression in human P2 chondrocytes, human P2 chondrocytes were exposed to cytochalasin D. Cytochalasin D decreased COL1 gene expression in human passaged chondrocytes. Furthermore, culture of bovine P2 chondrocytes in 3D culture on porous bone substitute resulted in actin depolymerization, which correlated with decreased expression of COL1 and proliferation molecules. In 3D cultures, aggrecan gene expression was increased by cytochalasin D treatment and COL1 was further decreased. These results reveal that actin polymerization

  19. Metabolic Effects of Avocado/Soy Unsaponifiables on Articular Chondrocytes

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    Louis Lippiello

    2008-01-01

    Full Text Available Avocado/soy unsaponifiable (ASU components are reported to have a chondroprotective effect by virtue of anti-inflammatory and proanabolic effects on articular chondrocytes. The identity of the active component(s remains unknown. In general, sterols, the major component of unsaponifiable plant material have been demonstrated to be anti-inflammatory in vitro and in animal models. These studies were designed to clarify whether the sterol content of ASU preparations were the primary contributors to biological activity in articular chondrocytes. ASU samples were analyzed by high pressure liquid chromatography (HPLC and GC mass spectrometry. The sterol content was normalized between diverse samples prior to in vitro testing on bovine chondrocytes. Anabolic activity was monitored by uptake of 35-sulfate into proteoglycans and quantitation of labeled hydroxyproline and proline content after incubation with labeled proline. Anti-inflammatory activity was assayed by measuring reduction of interleukin-1 (IL-1-induced synthesis of PGE2 and metalloproteases and release of label from tissue prelabeled with S-35.All ASU samples exerted a similar time-dependent up-regulation of 35-sulfate uptake in bovine cells reaching a maximum of greater than 100% after 72 h at sterol doses of 1–10 μg/ml. Non-collagenous protein (NCP and collagen synthesis were similarly up-regulated. All ASU were equally effective in dose dependently inhibiting IL-1-induced MMP-3 activity (23–37%, labeled sulfate release (15–23% and PGE2 synthesis (45–58%. Up-regulation of glycosaminoglycan and collagen synthesis and reduction of IL-1 effects in cartilage are consistent with chondroprotective activity. The similarity of activity of ASU from diverse sources when tested at equal sterol levels suggests sterols are important for biologic effects in articular chondrocytes.

  20. Metabolic Effects of Avocado/Soy Unsaponifiables on Articular Chondrocytes

    Science.gov (United States)

    Nardo, Joseph V.; Harlan, Robert; Chiou, Tiffany

    2008-01-01

    Avocado/soy unsaponifiable (ASU) components are reported to have a chondroprotective effect by virtue of anti-inflammatory and proanabolic effects on articular chondrocytes. The identity of the active component(s) remains unknown. In general, sterols, the major component of unsaponifiable plant material have been demonstrated to be anti-inflammatory in vitro and in animal models. These studies were designed to clarify whether the sterol content of ASU preparations were the primary contributors to biological activity in articular chondrocytes. ASU samples were analyzed by high pressure liquid chromatography (HPLC) and GC mass spectrometry. The sterol content was normalized between diverse samples prior to in vitro testing on bovine chondrocytes. Anabolic activity was monitored by uptake of 35-sulfate into proteoglycans and quantitation of labeled hydroxyproline and proline content after incubation with labeled proline. Anti-inflammatory activity was assayed by measuring reduction of interleukin-1 (IL-1)-induced synthesis of PGE2 and metalloproteases and release of label from tissue prelabeled with S-35.All ASU samples exerted a similar time-dependent up-regulation of 35-sulfate uptake in bovine cells reaching a maximum of greater than 100% after 72 h at sterol doses of 1–10 μg/ml. Non-collagenous protein (NCP) and collagen synthesis were similarly up-regulated. All ASU were equally effective in dose dependently inhibiting IL-1-induced MMP-3 activity (23–37%), labeled sulfate release (15–23%) and PGE2 synthesis (45–58%). Up-regulation of glycosaminoglycan and collagen synthesis and reduction of IL-1 effects in cartilage are consistent with chondroprotective activity. The similarity of activity of ASU from diverse sources when tested at equal sterol levels suggests sterols are important for biologic effects in articular chondrocytes. PMID:18604259

  1. Epigenetic Regulation of Chondrocyte Catabolism and Anabolism in Osteoarthritis

    OpenAIRE

    Kim, Hyeonkyeong; Kang, Donghyun; Cho, Yongsik; Kim, Jin-Hong

    2015-01-01

    Osteoarthritis (OA) is one of the most prevalent forms of joint disorder, associated with a tremendous socioeconomic burden worldwide. Various non-genetic and lifestyle-related factors such as aging and obesity have been recognized as major risk factors for OA, underscoring the potential role for epigenetic regulation in the pathogenesis of the disease. OA-associated epigenetic aberrations have been noted at the level of DNA methylation and histone modification in chondrocytes. These epigenet...

  2. Doublecortin May Play a Role in Defining Chondrocyte Phenotype

    Directory of Open Access Journals (Sweden)

    Dongxia Ge

    2014-04-01

    Full Text Available Embryonic development of articular cartilage has not been well understood and the role of doublecortin (DCX in determination of chondrocyte phenotype is unknown. Here, we use a DCX promoter-driven eGFP reporter mouse model to study the dynamic gene expression profiles in mouse embryonic handplates at E12.5 to E13.5 when the condensed mesenchymal cells differentiate into either endochondral chondrocytes or joint interzone cells. Illumina microarray analysis identified a variety of genes that were expressed differentially in the different regions of mouse handplate. The unique expression patterns of many genes were revealed. Cytl1 and 3110032G18RIK were highly expressed in the proximal region of E12.5 handplate and the carpal region of E13.5 handplate, whereas Olfr538, Kctd15, and Cited1 were highly expressed in the distal region of E12.5 and the metacarpal region of E13.5 handplates. There was an increasing gradient of Hrc expression in the proximal to distal direction in E13.5 handplate. Furthermore, when human DCX protein was expressed in human adipose stem cells, collagen II was decreased while aggrecan, matrilin 2, and GDF5 were increased during the 14-day pellet culture. These findings suggest that DCX may play a role in defining chondrocyte phenotype.

  3. Growth differentiation factor-5 stimulates the growth and anabolic metabolism of articular chondrocytes

    Institute of Scientific and Technical Information of China (English)

    Xu Peng; Guo Xiong; Yao Jianfeng; Zhang Yingang; Klaus von der Mark

    2005-01-01

    Objective: To observe the effect of growth differentiation factor-5 (GDF-5) on the growth and anabolic metabolism of articular chondrocytes. Methods: The articular chondrocytes isolated from rats were treated with various concentrations of rmGDF-5, and the growth of chondrocytes measured by MTT assay, the cellular cartilage matrices formation detected sulfated glycosaminoglycan by Alcian blue staining and type Ⅱ collagen by RT-PCR,the collagen phenotypic expression of chondrocytes detected by immunofluorescence. Results: After 7 days culture,MTT assay showed that GDF-5 enhanced the growth of chondrocytes in a dose-dependent manner, RT-PCR showed that GDF-5 clearly induced the synthesis of type Ⅱ collagen because of the col2a1 mRNA band more and more strong in a dose-dependent. Chondrocytes were cultured with GDF-5 for 14 days, the intensity of Alcian blue staining was greatly enhanced, especially, at a high concentration of 1000ng/ml, and GDF-5 enhanced the accumulation of the Alcian blue-stainable material in a concentration-dependent manner and in a does-dependent manner. Chondrocytes were cultured with GDF-5 for 21 days, immunofluorescent staining of type Ⅱ collagen was clear, the type Ⅰ and X collagen were negative. Conclusion: GDF-5 enhanced the growth of mature articular chondrocytes, and stimulated the cellular cartilage matrices formation, but did not change the collagen phenotypic expression of chondrocytes in mono-layer culture.

  4. Chondrogenic differentiation of mouse embryonic stem cells promoted by mature chondrocytes

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    In order to direct embryonic stem (ES) cells to differentiate into chondrocytes, a chondrogenic envi-ronment provided by mature chondrocytes was investigated. Flk-1 positive cells sorted from pre-differentiated mouse ES cells were mixed with adult porcine articular chondrocytes, seeded on biodegradable scaffolds, and then implanted subcutaneously into nude mice. The cell-scaffold com-plexes formed cartilage tissues after 4 weeks, which was demonstrated by histology and anti-type II collagen antibody staining. Positive staining of mouse Major Histocompatibility Complex class I molecules confirmed that part of the chondrocytes were derived from mouse ES cells. The current study established a new approach for directing ES cell differentiation.

  5. Chondrogenic differentiation of mouse embryonic stem cells promoted by mature chondrocytes

    Institute of Scientific and Technical Information of China (English)

    XIE Feng; ZHANG WenJie; CHEN FanFan; ZHOU GuangDong; CUI Lei; LIU Wei; CAO YiLin

    2008-01-01

    In order to direct embryonic stem (ES) cells to differentiate into chondrocytes, a chondrogenic envi-ronment provided by mature chondrocytes was investigated. FIk-1 positive cells sorted from pre-differentiated mouse ES cells were mixed with adult porcine articular chondrocytes, seeded on biodegradable scaffolds, and then implanted subcutaneously into nude mice. The cell-scaffold com-plexes formed cartilage tissues after 4 weeks, which was demonstrated by histology and anti-type Ⅱ collagen antibody staining. Positive staining of mouse Major Histocompatibility Complex class Ⅰ molecules confirmed that part of the chondrocytes were derived from mouse ES cells. The current study established a new approach for directing ES cell differentiation.

  6. Transforming the Terminal Associates of Applied Science into a Four-Year Degree: A Win-Win Situation for Students, Community Colleges, Universities, and Businesses

    Science.gov (United States)

    Batts, David L.; Pagliari, Leslie R.

    2013-01-01

    Associates of Applied Science (AAS) degrees were once considered terminal degrees and were developed for people seeking technical skills to join the workforce. This paper discusses the transformation from a transferable degree into technical four-year baccalaureate degree. It also discusses survey results of students currently in a degree…

  7. A Comparison of Academic Administrators and Enrollment Managers' Perceptions of Undergraduate Enrollment Management Functions at a Subset of Four-Year Public Institutions

    Science.gov (United States)

    Cesarini, Lisa McHugh

    2011-01-01

    The purpose of this study was to examine perceptions of various enrollment management functions at a subset of four-year public institutions. Specifically, this study compared perceptions of academic administrators with enrollment managers as they related to the availability, need, and effectiveness of certain enrollment management functions. In…

  8. Prognosis and Continuity of Child Mental Health Problems from Preschool to Primary School: Results of a Four-Year Longitudinal Study

    Science.gov (United States)

    Beyer, Thomas; Postert, Christian; Muller, Jorg M.; Furniss, Tilman

    2012-01-01

    In a four-year longitudinal study, changes in and continuity of behavioral and emotional problems were examined in 814 subjects from kindergarten to primary school. Mental health problems were assessed by means of the Child Behavior Checklist (CBCL). The distribution of the CBCL broadband groups revealed a high level of continuity of internalizing…

  9. An Exploratory Study on the Purpose, Structure, Format and Use of Syllabi at a Midwest Four-Year Undergraduate Private University

    Science.gov (United States)

    Fink, Susan Jo Breakenridge

    2011-01-01

    The purpose of this study was to explore how instructors at a mid-sized Midwest four-year undergraduate private university view the purpose, structure, format and use of their course syllabi. The theory of structural functionalism and a quantitative research approach were employed. A group administration approach was used to distribute the paper…

  10. Examining Factors of Acculturative Stress on International Students as They Affect Utilization of Campus-Based Health and Counseling Services at Four-Year Public Universities in Ohio

    Science.gov (United States)

    Hofmann, Paul N.

    2010-01-01

    This study examined factors of acculturative stress experienced by international students as they affect utilization of campus-based health and counseling services. Eight hundred thirty-eight international students studying at 11 four-year public institutions in the State of Ohio were surveyed to determine how frequently they had experienced 20…

  11. Potholes on the Road to College: High School Effects in Shaping Urban Students' Participation in College Application, Four-Year College Enrollment, and College Match

    Science.gov (United States)

    Roderick, Melissa; Coca, Vanessa; Nagaoka, Jenny

    2011-01-01

    This article examines the extent to which indicators of the college-going climate of urban high schools are associated with students' application to, enrollment in, and choice among four-year colleges. The investigators examine two mechanisms by which high schools may shape college enrollment among low-income students in an urban school system:…

  12. Women in Leadership: Factors That Affect the Achievement of Women in Higher Education Administration at Four-Year Public and Private Universities in Texas

    Science.gov (United States)

    Ramirez, Dawn Marie

    2012-01-01

    The purpose of this quantitative study was to examine the factors that affect women administrators in higher education at four-year public and private universities in Texas. By comparing private and public universities, the research provided an assessment of similarities and differences of the factors impacting achievement of women in higher…

  13. Chondrocyte Morphology in Stiff and Soft Agarose Gels and the Influence of Fetal Calf Serum.

    Science.gov (United States)

    Karim, Asima; Hall, Andrew C

    2017-05-01

    Changes to chondrocyte volume/morphology may have deleterious effects on extracellular matrix (ECM) metabolism potentially leading to cartilage deterioration and osteoarthritis (OA). The factors controlling chondrocyte properties are poorly understood, however, pericellular matrix (PCM) weakening may be involved. We have studied the density, volume, morphology, and clustering of cultured bovine articular chondrocytes within stiff (2% w/v) and soft (0.2% w/v) three-dimensional agarose gels. Gels with encapsulated chondrocytes were cultured in Dulbecco's Modified Eagle's Medium (DMEM; fetal calf serum (FCS) 1-10%;380 mOsm) for up to 7 days. Chondrocytes were fluorescently labeled after 1, 3, and 7 days with 5-chloromethylfluorescein-diacetate (CMFDA) and propidium iodide (PI) or 1,5-bis{[2-(di-methylamino)ethyl]amino}-4,8-dihydroxyanthracene-9,10-dione (DRAQ5) to identify cytoplasmic space or DNA and imaged by confocal laser scanning microscopy (CLSM). Chondrocyte density, volume, morphology, and clustering were quantified using Volocity™ software. In stiff gels after 7 d with 10% FCS, chondrocyte density remained unaffected and morphology was relatively normal with occasional cytoplasmic processes. However, in soft gels by day 1, chondrocyte volume increased (P = 0.0058) and by day 7, density increased (P = 0.0080), along with the percentage of chondrocytes of abnormal morphology (P stiff gels. FCS exacerbated changes to density (P stiffness and/or increased FCS concentrations promoted chondrocyte proliferation and clustering, increased cell volume, and stimulated abnormal morphology, producing similar changes to those occurring in OA. The increased penetration of factors in FCS into soft gels may be important in the development of these abnormal chondrocyte properties. J. Cell. Physiol. 232: 1041-1052, 2017. © 2016 Wiley Periodicals, Inc.

  14. Autophagy protects end plate chondrocytes from intermittent cyclic mechanical tension induced calcification.

    Science.gov (United States)

    Xu, Hong-guang; Yu, Yun-fei; Zheng, Quan; Zhang, Wei; Wang, Chuang-dong; Zhao, Xiao-yn; Tong, Wen-xue; Wang, Hong; Liu, Ping; Zhang, Xiao-ling

    2014-09-01

    Calcification of end plate chondrocytes is a major cause of intervertebral disc (IVD) degeneration. However, the underlying molecular mechanism of end plate chondrocyte calcification is still unclear. The aim of this study was to clarify whether autophagy in end plate chondrocytes could protect the calcification of end plate chondrocytes. Previous studies showed that intermittent cyclic mechanical tension (ICMT) contributes to the calcification of end plate chondrocytes in vitro. While autophagy serves as a cell survival mechanism, the relationship of autophagy and induced end plate chondrocyte calcification by mechanical tension in vitro is unknown. Thus, we investigated autophagy, the expression of the autophagy genes, Beclin-1 and LC3, and rat end plate chondrocyte calcification by ICMT. The viability of end plate chondrocytes was examined using the LIVE/DEAD viability/cytotoxicity kit. The reverse transcription-polymerase chain reaction and western blotting were used to detect the expression of Beclin-1; LC3; type I, II and X collagen; aggrecan; and Sox-9 genes. Immunofluorescent and fluorescent microscopy showed decreased autophagy in the 10- and 20-day groups loaded with ICMT. Additionally, Alizarin red and alkaline phosphatase staining detected the palpable calcification of end plate chondrocytes after ICMT treatment. We found that increased autophagy induced by short-term ICMT treatment was accompanied by an insignificant calcification of end plate chondrocytes. To the contrary, the suppressive autophagy inhibited by long-term ICMT was accompanied by a more significant calcification. The process of calcification induced by ICMT was partially resisted by increased autophagy activity induced by rapamycin, implicating that autophagy may prevent end plate chondrocyte calcification.

  15. Reduction of Environmental Temperature Mitigates Local Anesthetic Cytotoxicity in Bovine Articular Chondrocytes

    Directory of Open Access Journals (Sweden)

    Tarik Onur, Alexis Dang

    2014-09-01

    Full Text Available The purpose of this study was to assess whether reducing environmental temperature will lead to increased chondrocyte viability following injury from a single-dose of local anesthetic treatment. Bovine articular chondrocytes from weight bearing portions of femoral condyles were harvested and cultured. 96-well plates were seeded with 15,000 chondrocytes per well. Chondrocytes were treated with one of the following conditions: ITS Media, 1x PBS, 2% lidocaine, 0.5% bupivacaine, or 0.5% ropivacaine. Each plate was then incubated at 37°C, 23°C, or 4°C for one hour and then returned to media at 37°C. Chondrocyte viability was assessed 24 hours after treatment. Chondrocyte viability is presented as a ratio of the fluorescence of the treatment group over the average of the media group at that temperature (ratio ± SEM. At 37°C, lidocaine (0.35 ± 0.04 and bupivacaine (0.30 ± 0.05 treated chondrocytes show low cell viability when compared to the media (1.00 ± 0.03 control group (p < 0.001. Lidocaine treated chondrocytes were significantly more viable at 23°C (0.84 ± 0.08 and 4°C (0.86±0.085 than at 37°C (p < 0.001. Bupivacaine treated chondrocytes were significantly more viable at 4°C (0.660 ± 0.073 than at 37°C or 23°C (0.330 ± 0.069 (p < 0.001 and p = 0.002 respectively. Reducing the temperature from 37°C to 23°C during treatment with lidocaine increases chondrocyte viability following injury. Chondrocytes treated with bupivacaine can be rescued by reducing the temperature to 4°C.

  16. Human serum provided additional values in growth factors supplemented medium for human chondrocytes monolayer expansion and engineered cartilage construction.

    Science.gov (United States)

    Chua, K H; Aminuddin, B S; Fuzina, N H; Ruszymah, B H I

    2004-05-01

    We have previously formulated an optimized human chondrocytes growth medium based on 2% fetal bovine serum supplementation. For clinical usage, the animal serum must be replaced by patient own serum. We investigated the effects of human serum concentration for human nasal septum chondrocytes monolayer culture and cartilage reconstruction. Human serum demonstrated a dose dependent manner in promoting chondrocytes growth and cartilage engineering.

  17. Preservation of the chondrocyte's pericellular matrix improves cell-induced cartilage formation

    NARCIS (Netherlands)

    Vonk, L.A.; Doulabi, B.Z.; Huang, C.L.; Helder, M.N.; Everts, V.; Bank, R.A.

    2010-01-01

    The extracellular matrix surrounding chondrocytes within a chondron is likely to affect the metabolic activity of these cells. In this study we investigated this by analyzing protein synthesis by intact chondrons obtained from different types of cartilage and compared this with chondrocytes. Chondro

  18. Preservation of the Chondrocyte's Pericellular Matrix Improves Cell-Induced Cartilage Formation

    NARCIS (Netherlands)

    Vonk, Lucienne A.; Doulabi, Behrouz Zandieh; Huang, ChunLing; Helder, Marco N.; Everts, Vincent; Bank, Ruud A.

    2010-01-01

    The extracellular matrix surrounding chondrocytes within a chondron is likely to affect the metabolic activity of these cells. In this study we investigated this by analyzing protein synthesis by intact chondrons obtained from different types of cartilage and compared this with chondrocytes. Chondro

  19. Cultured articular chondrocytes sheets for partial thickness cartilage defects utilizing temperature-responsive culture dishes

    Directory of Open Access Journals (Sweden)

    N Kaneshiro

    2007-05-01

    Full Text Available The extracellular matrix (ECM of articular cartilage has several functions that are unique to joints. Although a technique for transplanting cultured chondrocytes has already been introduced, it is difficult to collect intact ECM when using enzymes to harvest samples. Temperature-responsive culture dishes have already been clinically applied in the fields of myocardial and corneal transplantation. Earlier studies have shown that a sheet of cultured cells with intact ECM and adhesive factors can be harvested using such culture dishes, which allow the surface properties of the dish to be reversibly altered by changing the temperature. Human chondrocytes were subjected to enzymatic digestion and then were seeded in temperature-responsive culture dishes. A sheet of chondrocytes was harvested by only reducing the temperature after the cultured cells reached confluency. A real-time PCR analysis of the chondrocyte sheets confirmed that type II collagen, aggrecan, and fibronectin were present. These results suggested that, although chondrocytes undergo dedifferentiation in a monolayer culture, multilayer chondrocyte sheets grown in a similar environment to that of three-dimensional culture may be able to maintain a normal phenotype. A histological examination suggested that multilayer chondrocyte sheets could thus prevent the loss of proteoglycans because the area covered by the sheets was well stained by safranin-O. The present experiments suggested that temperature-responsive culture dishes are useful for obtaining cultured chondrocytes, which may then be clinically employed as a substitute for periosteal patches because such sheets can be applied without a scaffold.

  20. Growth Differentiation Factor-5 Stimulates the Growth and Anabolic Metabolism of Articular Chondrocytes

    Institute of Scientific and Technical Information of China (English)

    Xu Peng; Yao Jianfeng; Guo Xiong; Zhang Yingang; Klaus von der Mark

    2009-01-01

    Objective: To observe the effect of growth differentiation factor-5 (GDF-5) on the growth and anabolic metabolism of articular chondrocytes. Methods: The articular chondrocytes isolated from rats were treated with various concentrations of rmGDF-5, and the growth of chondrocytes measured by MTr assay, the cellular cartilage matrices formation detected sulfated glycosaminoglycan by Alcian blue staining and type 11 collagen by RT-PCR, the collagen phenotypic expression of chondrocytes detected by immunofluorescence. Results: After 7 days culture, MTF assay showed that GDF-5 enhanced the growth of ehondrocytes in a dose-dependent manner, RT-PCR showed that GDF-5 clearly induced the synthesis of type Ⅱ collagen because of the colal mRNA band more and more strong in a dose-dependent. Chondrocytes were cultured with GDF-5 for 14 days, the intensity of Alcian blue staining was gready enhanced, especially, at a high concentration of 1000ng/ml, and GDF-5 enhanced the accumulation of the Alcian blue-stainable material in a concentration-dependent manner and in a does-dependent manner. Chondrocytes were cultured with GDF-5 for 21 days, immunofluorescent staining of type Ⅱ collagen was clear, the type Ⅰ and Ⅹ collagen were negative. Conclusion: GDF-5 enhanced the growth of mature articular chon-drocytes, and stimulated the cellular cartilage matrices formation, but did not change the collagen phenotypic ex-pression of chondrocytes in mono-layer culture.

  1. Effect of oxygen tension on adult articular chondrocytes in microcarrier bioreactor culture

    NARCIS (Netherlands)

    Malda, J.; Brink, van den P.; Meeuwse, P.; Grojec, M.; Martens, D.E.; Tramper, J.; Riesle, J.; Blitterswijk, van C.A.

    2004-01-01

    Tissue-engineering approaches for cartilage repair hold promise for the treatment of cartilage defects. Various methods to prevent or reduce dedifferentiation during chondrocyte expansion are currently under investigation. In the present study we evaluated the effect of oxygen on chondrocyte prolife

  2. Chondrocyte hypertrophy and osteoarthritis: role in initiation and progression of cartilage degeneration?

    NARCIS (Netherlands)

    Kraan, P.M. van der; Berg, W.B. van den

    2012-01-01

    OBJECTIVE: To review the literature on the role and regulation of chondrocyte terminal differentiation (hypertrophy-like changes) in osteoarthritis (OA) and to integrate this in a conceptual model of primary OA development. METHODS: Papers investigating chondrocyte terminal differentiation in human

  3. Modulation of Hyaluronan Synthesis by the Interaction between Mesenchymal Stem Cells and Osteoarthritic Chondrocytes

    Directory of Open Access Journals (Sweden)

    Eliane Antonioli

    2015-01-01

    Full Text Available Bone marrow mesenchymal stem cells (BM-MSCs are considered a good source for cellular therapy in cartilage repair. But, their potential to repair the extracellular matrix, in an osteoarthritic environment, is still controversial. In osteoarthritis (OA, anti-inflammatory action and extracellular matrix production are important steps for cartilage healing. This study examined the interaction of BM-MSC and OA-chondrocyte on the production of hyaluronan and inflammatory cytokines in a Transwell system. We compared cocultured BM-MSCs and OA-chondrocytes with the individually cultured controls (monocultures. There was a decrease in BM-MSCs cell count in coculture with OA-chondrocytes when compared to BM-MSCs alone. In monoculture, BM-MSCs produced higher amounts of hyaluronan than OA-chondrocytes and coculture of BM-MSCs with OA-chondrocytes increased hyaluronan production per cell. Hyaluronan synthase-1 mRNA expression was upregulated in BM-MSCs after coculture with OA-chondrocytes, whereas hyaluronidase-1 was downregulated. After coculture, lower IL-6 levels were detected in BM-MSCs compared with OA-chondrocytes. These results indicate that, in response to coculture with OA-chondrocytes, BM-MSCs change their behavior by increasing production of hyaluronan and decreasing inflammatory cytokines. Our results indicate that BM-MSCs per se could be a potential tool for OA regenerative therapy, exerting short-term effects on the local microenvironment even when cell:cell contact is not occurring.

  4. Direct measurement of TRPV4 and PIEZO1 activity reveals multiple mechanotransduction pathways in chondrocytes

    Science.gov (United States)

    Rocio Servin-Vences, M; Moroni, Mirko; Lewin, Gary R; Poole, Kate

    2017-01-01

    The joints of mammals are lined with cartilage, comprised of individual chondrocytes embedded in a specialized extracellular matrix. Chondrocytes experience a complex mechanical environment and respond to changing mechanical loads in order to maintain cartilage homeostasis. It has been proposed that mechanically gated ion channels are of functional importance in chondrocyte mechanotransduction; however, direct evidence of mechanical current activation in these cells has been lacking. We have used high-speed pressure clamp and elastomeric pillar arrays to apply distinct mechanical stimuli to primary murine chondrocytes, stretch of the membrane and deflection of cell-substrate contacts points, respectively. Both TRPV4 and PIEZO1 channels contribute to currents activated by stimuli applied at cell-substrate contacts but only PIEZO1 mediates stretch-activated currents. These data demonstrate that there are separate, but overlapping, mechanoelectrical transduction pathways in chondrocytes. DOI: http://dx.doi.org/10.7554/eLife.21074.001 PMID:28135189

  5. Cartilage homeoprotein 1, a homeoprotein selectively expressed in chondrocytes.

    OpenAIRE

    Zhao, G. Q.; Zhou, X.; Eberspaecher, H; Solursh, M; de Crombrugghe, B

    1993-01-01

    We identified a rat cDNA that encodes cartilage homeoprotein 1 (Cart-1). The deduced amino acid sequence of Cart-1 contains a paired-type homeodomain. Northern blot hybridization and RNase protection assay revealed that Cart-1 RNA was present at high levels in a well-differentiated rat chondrosarcoma tumor and in a cell line derived from this tumor. Cart-1 RNA was detected in primary mouse and rat chondrocytes but not in various fibroblasts including mouse 10T1/2 cells, NIH 3T3 cells, BALB 3T...

  6. An Analysis on Thailand's Economic Situation During Thaksin'Four- year Administration%他信执政四年来泰国经济形势分析

    Institute of Scientific and Technical Information of China (English)

    谢丽梅

    2005-01-01

    Since Thaksin' s four - year rule, Thailand' s economy has been steered out of the shadow of financial crisis, moved into a new turn of accelerating growth and is now walking towards the continuous and steady track thanks to ,from the internal factor, his Administration' s dual - track economic policy and from the external factor, the recovery of the international economic situation and the accelerating growth of Chinese economy.

  7. Cyclic Equibiaxial Tensile Strain Alters Gene Expression of Chondrocytes via Histone Deacetylase 4 Shuttling.

    Directory of Open Access Journals (Sweden)

    Chongwei Chen

    Full Text Available This paper aims to investigate whether equibiaxial tensile strain alters chondrocyte gene expression via controlling subcellular localization of histone deacetylase 4 (HDAC4.Murine chondrocytes transfected with GFP-HDAC4 were subjected to 3 h cyclic equibiaxial tensile strain (CTS, 6% strain at 0.25 Hz by a Flexcell® FX-5000™ Tension System. Fluorescence microscope and western blot were used to observe subcellular location of HDAC4. The gene expression was analyzed by real-time RT-PCR. The concentration of Glycosaminoglycans in culture medium was quantified by bimethylmethylene blue dye; Collagen II protein was evaluated by western blot. Cells phenotype was identified by immunohistochemistry. Cell viability was evaluated by live-dead cell detect kit. Okadaic acid, an inhibitor of HDAC4 nuclear relocation, was used to further validate whether HDAC4 nuclear relocation plays a role in gene expression in response to tension stimulation.87.5% of HDAC4 was located in the cytoplasm in chondrocytes under no loading condition, but it was relocated to the nucleus after CTS. RT-PCR analysis showed that levels of mRNA for aggrecan, collagen II, LK1 and SOX9 were all increased in chondrocytes subjected to CTS as compared to no loading control chondrocytes; in contrast, the levels of type X collagen, MMP-13, IHH and Runx2 gene expression were decreased in the chondrocytes subjected to CTS as compared to control chondrocytes. Meanwhile, CTS contributed to elevation of glycosaminoglycans and collagen II protein, but did not change collagen I production. When Okadaic acid blocked HDAC4 relocation from the cytoplasm to nucleus, the changes of the chondrocytes induced by CTS were abrogated. There was no chondrocyte dead detected in this study in response to CTS.CTS is able to induce HDAC4 relocation from cytoplasm to nucleus. Thus, CTS alters chondrocytes gene expression in association with the relocation of HDAC4 induced by CTS.

  8. Salvianolic acid B regulates gene expression and promotes cell viability in chondrocytes.

    Science.gov (United States)

    Yang, Xiaohong; Liu, Shaojie; Li, Siming; Wang, Pengzhen; Zhu, Weicong; Liang, Peihong; Tan, Jianrong; Cui, Shuliang

    2017-02-28

    Articular chondrocytes reside in lacunae distributed in cartilage responsible for the remodelling of the tissue with limited ability of damage repairing. The in vitro expanded chondrocytes enhanced by factors/agents to obtain large numbers of cells with strengthened phenotype are essential for successful repair of cartilage lesions by clinical cell implantation therapies. Because the salvianolic acid B (Sal B), a major hydrophilic therapeutic agent isolated from Salvia miltiorrhiza, has been widely used to treat diseases and able to stimulate activity of cells, this study examines the effects of Sal B on passaged chondrocytes. Chondrocytes were treated with various concentrations of Sal B in monolayer culture, their morphological properties and changes, and mitochondrial membrane potential were analysed using microscopic analyses, including cellular biochemical staining and confocal laser scanning microscopy. The proteins were quantified by BCA and Western blotting, and the transcription of genes was detected by qRT-PCR. The passaged chondrocytes treated with Sal B showed strengthened cellular synthesis and stabilized mitochondrial membrane potential with upregulated expression of the marker genes for chondrocyte phenotype, Col2-α1, Acan and Sox9, the key Wnt signalling molecule β-catenin and paracrine cytokine Cytl-1. The treatments using CYTL-1 protein significantly increased expression of Col2-α1 and Acan with no effect on Sox9, indicating the paracrine cytokine acts on chondrocytes independent of SOX9. Sal B has ultimately promoted cell growth and enhanced chondrocyte phenotype. The chondrocytes treated with pharmaceutical agent and cytokine in the formulated medium for generating large number of differentiated chondrocytes would facilitate the cell-based therapies for cartilage repair.

  9. Indian hedgehog signals independently of PTHrP to promote chondrocyte hypertrophy.

    Science.gov (United States)

    Mak, Kinglun Kingston; Kronenberg, Henry M; Chuang, Pao-Tien; Mackem, Susan; Yang, Yingzi

    2008-06-01

    Chondrocyte hypertrophy is an essential process required for endochondral bone formation. Proper regulation of chondrocyte hypertrophy is also required in postnatal cartilage homeostasis. Indian hedgehog (Ihh) and PTHrP signaling play crucial roles in regulating the onset of chondrocyte hypertrophy by forming a negative feedback loop, in which Ihh signaling regulates chondrocyte hypertrophy by controlling PTHrP expression. To understand whether there is a PTHrP-independent role of Ihh signaling in regulating chondrocyte hypertrophy, we have both activated and inactivated Ihh signaling in the absence of PTHrP during endochondral skeletal development. We found that upregulating Ihh signaling in the developing cartilage by treating PTHrP(-/-) limb explants with sonic hedgehog (Shh) protein in vitro, or overexpressing Ihh in the cartilage of PTHrP(-/-) embryos or inactivating patched 1 (Ptch1), a negative regulator of hedgehog (Hh) signaling, accelerated chondrocyte hypertrophy in the PTHrP(-/-) embryos. Conversely, when Hh signaling was blocked by cyclopamine or by removing Smoothened (Smo), a positive regulator of Hh signaling, chondrocyte hypertrophy was delayed in the PTHrP(-/-) embryo. Furthermore, we show that upregulated Hh signaling in the postnatal cartilage led to accelerated chondrocyte hypertrophy during secondary ossification, which in turn caused reduction of joint cartilage. Our results revealed a novel role of Ihh signaling in promoting chondrocyte hypertrophy independently of PTHrP, which is particularly important in postnatal cartilage development and homeostasis. In addition, we found that bone morphogenetic protein (Bmp) and Wnt/beta-catenin signaling in the cartilage may both mediate the effect of upregulated Ihh signaling in promoting chondrocyte hypertrophy.

  10. A four-year follow-up controlled study of stress response and symptom persistence in Brazilian children and adolescents with attention deficit disorder and hyperactivity (ADHD).

    Science.gov (United States)

    Palma, Sonia Maria Motta; Natale, Ana Carolina Motta Palma; Calil, Helena Maria

    2015-12-15

    This study evaluated children and adolescents with Attention Deficit Disorder andHyperactivity Disorder (ADHD), reassessing them at a four-year follow-up. Their cortisol response to a stress stimulus was measured twice. ADHD symptom persistence, development of comorbidities, and psychostimulant usage were also reassessed. The initial sample consisted of 38 ADHD patients and 38 healthy controls, age ranging 6-14. At the follow-up, there were 37 ADHD patients and 22 healthy controls, age ranging 10-18. ADHD was classified as persistent if the patients fulfilled all DSM IV criteria for syndromic or subthreshold or had functional impairment. Salivary cortisol samples were collected prior to the application of a cognitive stressor (Continuous Performance Test - CPT), and at three time intervals afterwards at baseline and at the follow-up. Their reassessment showed that 75% had persistent symptoms, psychiatric comorbidities (oppositional defiant and behavioral disorders), functional and academic impairement. Only seven patients were on medication. The ADHD group's cortisol levels were lower than those measured four years earlier, but cortisol concentrations were similar for both ADHD and control groups at the four-year follow-up. The cortisol results suggest that HPA axis reactivity could be a marker differentiating ADHD from ADHD with comorbidities.

  11. Expression of type I collagen and tenascin C is regulated by actin polymerization through MRTF in dedifferentiated chondrocytes.

    Science.gov (United States)

    Parreno, Justin; Raju, Sneha; Niaki, Mortah Nabavi; Andrejevic, Katarina; Jiang, Amy; Delve, Elizabeth; Kandel, Rita

    2014-10-16

    This study examined actin regulation of fibroblast matrix genes in dedifferentiated chondrocytes. We demonstrated that dedifferentiated chondrocytes exhibit increased actin polymerization, nuclear localization of myocardin related transcription factor (MRTF), increased type I collagen (col1) and tenascin C (Tnc) gene expression, and decreased Sox9 gene expression. Induction of actin depolymerization by latrunculin treatment or cell rounding, reduced MRTF nuclear localization, repressed col1 and Tnc expression, and increased Sox9 gene expression in dedifferentiated chondrocytes. Treatment of passaged chondrocytes with MRTF inhibitor repressed col1 and Tnc expression, but did not affect Sox9 expression. Our results show that actin polymerization regulates fibroblast matrix gene expression through MRTF in passaged chondrocytes.

  12. Study of cryopreservation of articular chondrocytes using the Taguchi method.

    Science.gov (United States)

    Lyu, Shaw-Ruey; Wu, Wei Te; Hou, Chien Chih; Hsieh, Wen-Hsin

    2010-04-01

    This study evaluates the effect of control factors on cryopreservation of articular cartilage chondrocytes using the Taguchi method. Freeze-thaw experiments based on the L(8)(2(7)) two-level orthogonal array of the Taguchi method are conducted, and ANOVA (analysis of variables) is adopted to determine the statistically significant control factors that affect the viability of the cell. Results show that the type of cryoprotectant, freezing rate, thawing rate, and concentration of cryoprotectant (listed in the order of influence) are the statistically significant control factors that affect the post-thaw viability. The end temperature and durations of the first and second stages of freezing do not affect the post-thaw viability. Within the ranges of the control factors studied in this work, the optimal test condition is found to be a freezing rate of 0.61+/-0.03 degrees C/min, a thawing rate of 126.84+/-5.57 degrees C/min, Me(2)SO cryoprotectant, and a cryoprotectant concentration of 10% (v/v) for maximum cell viability. In addition, this study also explores the effect of cryopreservation on the expression of type II collagen using immunocytochemical staining and digital image processing. The results show that the ability of cryopreserved chondrocytes to express type II collagen is reduced within the first five days of monolayer culture.

  13. PTHrP regulates chondrocyte maturation in condylar cartilage.

    Science.gov (United States)

    Rabie, A B M; Tang, G H; Xiong, H; Hägg, U

    2003-08-01

    PTHrP is a key factor regulating the pace of endochondral ossification during skeletal development. Mandibular advancement solicits a cascade of molecular responses in condylar cartilage. However, the pace of cellular maturation and its effects on condylar growth are still unknown. The purpose of this study was to evaluate the pattern of expression of PTHrP and correlate it to cellular dynamics of chondrocytes in condylar cartilage during natural growth and mandibular advancement. We fitted 35-day-old Sprague-Dawley rats with functional appliances. Experimental animals with matched controls were labeled with bromodeoxyuridine 3 days before their death, so that mesenchymal cell differentiation could be traced. Mandibular advancement increased the number of differentiated chondroblasts and subsequently increased the cartilage volume. Higher levels of PTHrP expression in experimental animals coincided with the slowing of chondrocyte hypertrophy. Thus, mandibular advancement promoted mesenchymal cell differentiation and triggered PTHrP expression, which retarded their further maturation to allow for more growth.

  14. Antiangiogenic treatment delays chondrocyte maturation and bone formation during limb skeletogenesis.

    Science.gov (United States)

    Yin, Melinda; Gentili, Chiara; Koyama, Eiki; Zasloff, Michael; Pacifici, Maurizio

    2002-01-01

    Hypertrophic chondrocytes have important roles in promoting invasion of cartilage by blood vessels and its replacement with bone. However, it is unclear whether blood vessels exert reciprocal positive influences on chondrocyte maturation and function. Therefore, we implanted beads containing the antiangiogenic molecule squalamine around humeral anlagen in chick embryo wing buds and monitored the effects over time. Fluorescence microscopy showed that the drug diffused from the beads and accumulated in humeral perichondrial tissues, indicating that these tissues were the predominant targets of drug action. Diaphyseal chondrocyte maturation was indeed delayed in squalamine-treated humeri, as indicated by reduced cell hypertrophy and expression of type X collagen, transferrin, and Indian hedgehog (Ihh). Although reduced in amount, Ihh maintained a striking distribution in treated and control humeri, being associated with diaphyseal chondrocytes as well as inner perichondrial layer. These decreases were accompanied by lack of cartilage invasion and tartrate-resistant acid phosphatase-positive (TRAP+) cells and a significant longitudinal growth retardation. Recovery occurred at later developmental times, when in fact expression in treated humeri of markers such as matrix metalloproteinase 9 (MMP-9) and connective tissue growth factor (CTGF) appeared to exceed that in controls. Treating primary cultures of hypertrophic chondrocytes and osteoblasts with squalamine revealed no obvious changes in cell phenotype. These data provide evidence that perichondrial tissues and blood vessels in particular influence chondrocyte maturation in a positive manner and may cooperate with hypertrophic chondrocytes in dictating the normal pace and location of the transition from cartilage to bone.

  15. Softening Substrates Promote Chondrocytes Phenotype via RhoA/ROCK Pathway.

    Science.gov (United States)

    Zhang, Tao; Gong, Tao; Xie, Jing; Lin, Shiyu; Liu, Yao; Zhou, Tengfei; Lin, Yunfeng

    2016-09-01

    Due to its evascular, aneural, and alymphatic conditions, articular cartilage shows extremely poor regenerative ability. Thus, directing chondrocyte toward a desired location and function by utilizing the mechanical cues of biomaterials is a promising approach for effective tissue regeneration. However, chondrocytes cultured on Petri dish will lose their typical phenotype which may lead to compromised results. Therefore, we fabricated polydimethylsiloxane (PDMS) materials with various stiffness as culture substrates. Cell morphology and focal adhesion of chondrocytes displayed significant changes. The cytoskeletal tension of the adherent cells observed by average myosin IIA fluorescent intensity increased as stiffness of the underlying substrates decreased, consistent with the alteration of chondrocyte phenotype in our study. Immunofluorescent images and q-PCR results revealed that chondrocyte cultured on soft substrates showed better chondrocyte functionalization by more type II collagen and aggrecan expression, related to the lowest mRNA level of Rac-1, RhoA, ROCK-1, and ROCK-2. Taken together, this work not only points out that matrix elasticity can regulate chondrocyte functionalization via RhoA/ROCK pathway, but also provides new prospect for biomechanical control of cell behavior in cell-based cartilage regeneration.

  16. Forty Four Years a King.

    Science.gov (United States)

    Minneapolis Public Schools, Minn. Dept. of Intergroup Education.

    This curriculum unit contains guidelines for Martin Luther King Jr. Day, January 15th. Items included in the guide are: a list of suggested activities, a short biography of Dr. King, excerpts from some of his speeches including the famous, "I Have A Dream" speech, a play, study questions, "A King's Journey", a game, and a story…

  17. Cartilage tissue engineering of nasal septal chondrocyte-macroaggregates in human demineralized bone matrix.

    Science.gov (United States)

    Liese, Juliane; Marzahn, Ulrike; El Sayed, Karym; Pruss, Axel; Haisch, Andreas; Stoelzel, Katharina

    2013-06-01

    Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates. The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. In conclusion, this study provides the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilage. The as-yet insufficient redifferentiation of the chondrocytes and the sporadic initiation of apoptosis will require further investigations.

  18. Effects of intermittent versus continuous parathyroid hormone administration on condylar chondrocyte proliferation and differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Qi; Wan, Qilong; Yang, Rongtao; Zhou, Haihua [The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China); Li, Zubing, E-mail: lizubing0827@163.com [The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China); Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Wuhan University, 237 Luoyu Road, Wuhan 430079 (China)

    2012-07-20

    Highlights: Black-Right-Pointing-Pointer Different PTH administration exerts different effects on condylar chondrocyte. Black-Right-Pointing-Pointer Intermittent PTH administration suppresses condylar chondrocyte proliferation. Black-Right-Pointing-Pointer Continuous PTH administration maintains condylar chondrocyte proliferating. Black-Right-Pointing-Pointer Intermittent PTH administration enhances condylar chondrocyte differentiation. -- Abstract: Endochondral ossification is a complex process involving chondrogenesis and osteogenesis regulated by many hormones and growth factors. Parathyroid hormone (PTH), one of the key hormones regulating bone metabolism, promotes osteoblast differentiation and osteogenesis by intermittent administration, whereas continuous PTH administration inhibits bone formation. However, the effects of PTH on chondrocyte proliferation and differentiation are still unclear. In this study, intermittent PTH administration presented enhanced effects on condylar chondrocyte differentiation and bone formation, as demonstrated by increased mineral nodule formation and alkaline phosphatase (ALP) activity, up-regulated runt-related transcription factor 2 (RUNX2), ALP, collagen type X (COL10a1), collagen type I (COL1a1), osteocalcin (OCN), bone sialoprotein (BSP), bone morphogenetic protein 2 (BMP2) and osterix (OSX) mRNA and/or protein expression. On the contrary, continuous PTH administration promoted condylar chondrocyte proliferation and suppressed its differentiation, as demonstrated by up-regulated collagen type II (COL2a1) mRNA expression, reduced mineral nodule formation and down-regulated expression of the mRNAs and/or proteins mentioned above. Our data suggest that PTH can regulate condylar chondrocyte proliferation and differentiation, depending on the type of PTH administration. These results provide new insight into the effects of PTH on condylar chondrocytes and new evidence for using local PTH administration to cure mandibular

  19. Prolonged application of high fluid shear to chondrocytes recapitulates gene expression profiles associated with osteoarthritis.

    Directory of Open Access Journals (Sweden)

    Fei Zhu

    Full Text Available BACKGROUND: Excessive mechanical loading of articular cartilage producing hydrostatic stress, tensile strain and fluid flow leads to irreversible cartilage erosion and osteoarthritic (OA disease. Since application of high fluid shear to chondrocytes recapitulates some of the earmarks of OA, we aimed to screen the gene expression profiles of shear-activated chondrocytes and assess potential similarities with OA chondrocytes. METHODOLOGY/PRINCIPAL FINDINGS: Using a cDNA microarray technology, we screened the differentially-regulated genes in human T/C-28a2 chondrocytes subjected to high fluid shear (20 dyn/cm(2 for 48 h and 72 h relative to static controls. Confirmation of the expression patterns of select genes was obtained by qRT-PCR. Using significance analysis of microarrays with a 5% false discovery rate, 71 and 60 non-redundant transcripts were identified to be ≥2-fold up-regulated and ≤0.6-fold down-regulated, respectively, in sheared chondrocytes. Published data sets indicate that 42 of these genes, which are related to extracellular matrix/degradation, cell proliferation/differentiation, inflammation and cell survival/death, are differentially-regulated in OA chondrocytes. In view of the pivotal role of cyclooxygenase-2 (COX-2 in the pathogenesis and/or progression of OA in vivo and regulation of shear-induced inflammation and apoptosis in vitro, we identified a collection of genes that are either up- or down-regulated by shear-induced COX-2. COX-2 and L-prostaglandin D synthase (L-PGDS induce reactive oxygen species production, and negatively regulate genes of the histone and cell cycle families, which may play a critical role in chondrocyte death. CONCLUSIONS/SIGNIFICANCE: Prolonged application of high fluid shear stress to chondrocytes recapitulates gene expression profiles associated with osteoarthritis. Our data suggest a potential link between exposure of chondrocytes/cartilage to abnormal mechanical loading and the pathogenesis

  20. Precipitant induced porosity augmentation of polystyrene preserves the chondrogenicity of human chondrocytes.

    Science.gov (United States)

    Joergensen, Natasja L; Foldager, Casper B; Le, Dang Q S; Lind, Martin; Lysdahl, Helle

    2016-12-01

    Cells constantly sense and receive chemical and physical signals from neighboring cells, interstitial fluid, and extracellular matrix, which they integrate and translate into intracellular responses. Thus, the nature of the surface on which cells are cultured in vitro plays an important role for cell adhesion, proliferation, and differentiation. Autologs chondrocyte implantation is considered the treatment of choice for larger cartilage defects in the knee. To obtain a sufficient number of chondrocytes for implantation multiple passaging is often needed, which raises concerns about the changes in the chondrogenic phenotype. In the present study, we analyzed the effect at cellular and molecular level of precipitant induced porosity augmentation (PIPA) of polystyrene surfaces on proliferation and differentiation of human chondrocytes. Human chondrocytes were isolated from healthy patients undergoing anterior cruciate ligament reconstruction and cultured on PIPA modified polystyrene surfaces. Microscopical analysis revealed topographically arranged porosity with micron pores and nanometer pits. Chondrocytes cultured on PIPA surfaces revealed no difference in cell viability and proliferation, but gene- and protein expressions of collagen type II were pronounced in the first passage of chondrocytes when compared to chondrocytes cultured on control surfaces. Additionally, an analysis of 40 kinases revealed that chondrocytes expanded on PIPA caused upregulated PI3K/mTOR pathway activation and inhibition of mTORC1 resulted in reduced sGAG synthesis. These findings indicate that PIPA modified polystyrene preserved the chondrogenicity of expanded human chondrocytes at gene and protein levels, which clinically may be attractive for the next generation of cell-culture surfaces for ex vivo cell growth. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3073-3081, 2016.

  1. [Chickenpox, burns and grafts].

    Science.gov (United States)

    Rojas Zegers, J; Fidel Avendaño, L

    1979-01-01

    An outbreak of chickenpox that occurred at the Burns Repair Surgery Unit, Department of Children's Surgery, Hospital R. del Río, between June and November, 1975, is reported. 27 cases of burned children were studied, including analysis of correlations of the stages and outcome of the disease (varicela), the trauma (burns) and the graft (repair surgery). As a result, the authors emphasize the following findings: 1. Burns and their repair are not aggravating factors for varicella. In a small number of cases the exanthema looked more confluent in the graft surgical areas and in the first degree burns healing spontaneously. 2. Usually there was an uneventful outcome of graft repair surgery on a varicella patient, either during the incubation period, the acme or the convalescence. 3. The fact that the outmost intensity of secondary viremia of varicella occurs before the onset of exanthemia, that is, during the late incubation period, is confirmed.

  2. Alveolar bone grafting

    Directory of Open Access Journals (Sweden)

    Lilja Jan

    2009-10-01

    Full Text Available In patients with cleft lip and palate, bone grafting in the mixed dentition in the residual alveolar cleft has become a well-established procedure. The main advantages can be summarised as follows: stabilisation of the maxillary arch; facilitation of eruption of the canine and sometimes facilitation of the lateral incisor eruption; providing bony support to the teeth adjacent to the cleft; raising the alar base of the nose; facilitation of closure of an oro-nasal fistula; making it possible to insert a titanium fixture in the grafted site and to obtain favourable periodontal conditions of the teeth within and adjacent to the cleft. The timing of the ABG surgery take into consideration not only eruption of the canine but also that of the lateral incisor, if present. The best time for bone grafting surgery is when a thin shell of bone still covers the soon erupting lateral incisor or canine tooth close to the cleft.

  3. Arteriovenous shunt graft ulceration with sinus and graft epithelialization

    Directory of Open Access Journals (Sweden)

    Pooja Singhal

    2015-03-01

    Full Text Available Arteriovenous fistula and grafts are used as access sites for patients with chronic kidney disease and are prone for complications. Stent grafts are used to treat access site complications. We report a rare and unusual finding of epithelialization of the sinus tract and the lumen of a polytetrafluoroethylene graft, following ulceration of the overlying skin.

  4. Chitosan-graft-polyethylenimine/DNA nanoparticles as novel non-viral gene delivery vectors targeting osteoarthritis.

    Science.gov (United States)

    Lu, Huading; Dai, Yuhu; Lv, Lulu; Zhao, Huiqing

    2014-01-01

    The development of safe and efficient gene carriers is the key to the clinical success of gene therapy. The present study was designed to develop and evaluate the chitosan-graft-polyethylenimine (CP)/DNA nanoparticles as novel non-viral gene vectors for gene therapy of osteoarthritis. The CP/DNA nanoparticles were produced through a complex coacervation of the cationic polymers with pEGFP after grafting chitosan (CS) with a low molecular weight (Mw) PEI (Mw = 1.8 kDa). Particle size and zeta potential were related to the weight ratio of CP:DNA, where decreases in nanoparticle size and increases in surface charge were observed as CP content increased. The buffering capacity of CP was significantly greater than that of CS. The transfection efficiency of CP/DNA nanoparticles was similar with that of the Lipofectamine™ 2000, and significantly higher than that of CS/DNA and PEI (25 kDa)/DNA nanoparticles. The transfection efficiency of the CP/DNA nanoparticles was dependent on the weight ratio of CP:DNA (w/w). The average cell viability after the treatment with CP/DNA nanoparticles was over 90% in both chondrocytes and synoviocytes, which was much higher than that of PEI (25 kDa)/DNA nanoparticles. The CP copolymers efficiently carried the pDNA inside chondrocytes and synoviocytes, and the pDNA was detected entering into nucleus. These results suggest that CP/DNA nanoparticles with improved transfection efficiency and low cytotoxicity might be a safe and efficient non-viral vector for gene delivery to both chondrocytes and synoviocytes.

  5. Rapid Chondrocyte Isolation for Tissue Engineering Applications: The Effect of Enzyme Concentration and Temporal Exposure on the Matrix Forming Capacity of Nasal Derived Chondrocytes

    Directory of Open Access Journals (Sweden)

    Srujana Vedicherla

    2017-01-01

    Full Text Available Laboratory based processing and expansion to yield adequate cell numbers had been the standard in Autologous Disc Chondrocyte Transplantation (ADCT, Allogeneic Juvenile Chondrocyte Implantation (NuQu®, and Matrix-Induced Autologous Chondrocyte Implantation (MACI. Optimizing cell isolation is a key challenge in terms of obtaining adequate cell numbers while maintaining a vibrant cell population capable of subsequent proliferation and matrix elaboration. However, typical cell yields from a cartilage digest are highly variable between donors and based on user competency. The overall objective of this study was to optimize chondrocyte isolation from cartilaginous nasal tissue through modulation of enzyme concentration exposure (750 and 3000 U/ml and incubation time (1 and 12 h, combined with physical agitation cycles, and to assess subsequent cell viability and matrix forming capacity. Overall, increasing enzyme exposure time was found to be more detrimental than collagenase concentration for subsequent viability, proliferation, and matrix forming capacity (sGAG and collagen of these cells resulting in nonuniform cartilaginous matrix deposition. Taken together, consolidating a 3000 U/ml collagenase digest of 1 h at a ratio of 10 ml/g of cartilage tissue with physical agitation cycles can improve efficiency of chondrocyte isolation, yielding robust, more uniform matrix formation.

  6. Triclocarban, triclosan and its transformation product methyl triclosan in native earthworm species four years after a commercial-scale biosolids application.

    Science.gov (United States)

    Macherius, André; Lapen, David R; Reemtsma, Thorsten; Römbke, Jörg; Topp, Edward; Coors, Anja

    2014-02-15

    Triclocarban (TCC), triclosan (TCS) and methyl triclosan (Me-TCS) were detected in soil and the native population of earthworms of an agricultural field in Ottawa, Canada, about four years after a commercial-scale application of biosolids. In soil that received biosolids, TCC and TCS were detected at median concentrations of 13.0 and 1.5 ng/g soil (d.w.), respectively, while Me-TCS, the transformation product of triclosan, was detected at a six-fold higher median concentration than its precursor. In earthworms collected at the biosolids-amended field-plot about four years post application, Me-TCS was also detected at higher concentrations (26 to 114 ng/g tissue d.w.) than TCS (16-51 ng/g) and TCC (4-53 ng/g). These data provide evidence that not only parent compounds but also their transformation products need to be considered in faunal bioaccumulation studies. Moreover, the preliminary results for pooled earthworm samples from different ecological groups suggest that the degree of bioaccumulation of biosolids-associated contaminants may depend on the habitat and feeding behavior of the organisms.

  7. Lifestyle and Dietary Factors Associated with the Evolution of Cardiometabolic Risk over Four Years in West-African Adults: The Benin Study

    Directory of Open Access Journals (Sweden)

    Charles Sossa

    2013-01-01

    Full Text Available Aim. To assess in adults from Benin changes in cardiometabolic risk (CMR using both the Framingham risk score (FRS and metabolic syndrome (MetS and to examine the effects of diet, and lifestyles, controlling for location and socioeconomic status. Methods. Apparently healthy subjects (n=541 aged 25–60 years and randomly selected in the largest city, a small town, and rural areas were included in the four-year longitudinal study. Along with CMR factors, socioeconomic, diet and lifestyle data were collected in individual interviews. A food score based on consumption frequency of four “sentinel” food groups (meat and poultry, dairy, eggs, and vegetables was developed. Lifestyle included physical activity, alcohol and tobacco use. Education and income (proxy were the socioeconomic variables. Results. Among the subjects with four-year follow-up data (n=416, 13.5% were at risk at baseline, showing MetS or FRS ≥ 10%. The incidence of MetS and FRS ≥ 10% during follow-up was 8.2% and 5%, respectively. CMR deteriorated in 21% of subjects. Diet and lifestyle mediated location and income effects on CMR evolution. Low food scores and inactivity increased the likelihood of CMR deterioration. Conclusion. Combining MetS and FRS might be appropriate for surveillance purposes in order to better capture CMR and inform preventive measures.

  8. bFGF influences human articular chondrocyte differentiation

    DEFF Research Database (Denmark)

    Schmal, H; Zwingmann, J; Fehrenbach, M

    2007-01-01

    FGF concentrations in supernatants of primary human articular chondrocytes peaked immediately after isolation and then declined. In a dose-dependent manner, bFGF enhanced cell amplification and viability. BFGF induced a decrease in the apoptotic cell population, while the number of proliferating cells remained...... in monolayer. bFGF-dependent cell proliferation, production of collagen type II and aggrecan were monitored 10 days after isolation. Furthermore, effect of bFGF on cell cycle, cell morphology, and mRNA expression of integrins and chondrogenic markers determined by real time PCR were analyzed. RESULTS: b...... unchanged. Supplementation of cell culture with bFGF reduced collagen type II mRNA by 49%, but increased expression of the integrin alpha(2) by 70%. bFGF did not significantly regulate the integrins alpha(1), alpha(5), alpha(10), alpha(v) and type I collagen. bFGF reduced the amount of collagen type II...

  9. Effects of Cryoprotective Agents on the Bovine Articular Chondrocyte Viability

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Cryopreservation is the process of choice for long term preservation of cells and tissues. In this study, the effects of cryoprotective agents, dimethyl sulfoxide(DMSO), glycerol and 1,2-propanediol on the bovine articular chondrocyte viability were examined experimentally. The CPA was added at the concentrations of 0. 6. 0.9, 1.2 and 1.5 mol/I and at 4℃ and 37℃ and removed at 37℃ in one-step. CPA stepwise addition and removal at 0. 6 and 1. 2 mol/L and at 37℃ was also tested as an alternative protocol. Cell volume excursion during DMSO addition and removal was estimated and correlated well with cell survival rates. Solution makeup affects cell survival rate and a stepwise protocol can improve the cell survival rates significantly.

  10. Second-generation autologous chondrocyte transplantation: MRI findings and clinical correlations at a minimum 5-year follow-up

    Energy Technology Data Exchange (ETDEWEB)

    Kon, E. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Di Martino, A., E-mail: a.dimartino@biomec.ior.it [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Filardo, G. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Tetta, C.; Busacca, M. [Radiology, Rizzoli Orthopaedic Institute, Bologna (Italy); Iacono, F. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy); Delcogliano, M. [Orthopaedic Departement San Carlo di Nancy Hospital, Rome (Italy); Albisinni, U. [Radiology, Rizzoli Orthopaedic Institute, Bologna (Italy); Marcacci, M. [Biomechanics Laboratory, III Clinic, Rizzoli Orthopaedic Institute, Via Di Barbiano 1/10, 40136 Bologna (Italy)

    2011-09-15

    Objective: To evaluate the clinical outcome of hyaluronan-based arthroscopic autologous chondrocyte transplantation at a minimum of 5 years of follow-up and to correlate it with the MRI evaluation parameters. Methods: Fifty consecutive patients were included in the study and evaluated clinically using the Cartilage Standard Evaluation Form as proposed by ICRS and the Tegner score. Forty lesions underwent MRI evaluation at a minimum 5-year follow-up. For the description and evaluation of the graft, we employed the MOCART-scoring system. Results: A statistically significant improvement in all clinical scores was observed at 2 and over 5 years. The total MOCART score and the signal intensity (3D-GE-FS) of the repair tissue were statistically correlated to the IKDC subjective evaluation. Larger size of the treated cartilage lesions had a negative influence on the degree of defect repair and filling, the integration to the border zone and the subchondral lamina integrity, whereas more intensive sport activity had a positive influence on the signal intensity of the repair tissue, the repair tissue surface, and the clinical outcome. Conclusion: Our findings confirm the durability of the clinical results obtained with Hyalograft C and the usefulness of MRI as a non-invasive method for the evaluation of the repaired tissue and the outcome after second-generation autologous transplantation over time.

  11. Primary renal graft thrombosis

    NARCIS (Netherlands)

    Bakir, N; Sluiter, WJ; Ploeg, RJ; van Son, WJ; Tegzess, Adam

    1996-01-01

    Background. Renal allograft thrombosis is a serious complication of kidney transplantation that ultimately leads to graft loss. Its association with acute and hyperacute rejection is well documented; however, in a large proportion of patients the precise cause remains obscure. The exact incidence an

  12. Human platelet releasates combined with polyglycolic acid scaffold promote chondrocyte differentiation and phenotypic maintenance

    Indian Academy of Sciences (India)

    Giulia Bernardini; Federico Chellini; Bruno Frediani; Adriano Spreafico; Annalisa Santucci

    2015-03-01

    In the present study, we aimed to demonstrate the differentiating properties of platelet-rich plasma releasates (PRPr) on human chondrocytes seeded on a polygtlycolic acid (PGA) 3D scaffold. Gene expression and biochemical analysis were carried out to assess the improved quality of our PGA-based cartilage constructs supplemented with PRPr. We observed that the use of PRPr as cell cultures supplementation to PGA-chondrocyte constructs may promote chondrocyte differentiation, and thus may contribute to maintaining the chondrogenic phenotype longer than conventional supplementation by increasing high levels of important chondrogenic markers (e.g. sox9, aggrecan and type II collagen), without induction of type I collagen. Moreover, our constructs were analysed for the secretion and deposition of important ECM molecules (sGAG, type II collagen, etc.). Our results indicate that PRPr supplementation may synergize with PGA-based scaffolds to stimulate human articular chondrocyte differentiation, maturation and phenotypic maintenance.

  13. Autologous chondrocyte implantation for treatment of cartilage defects of the knee

    DEFF Research Database (Denmark)

    Jungmann, Pia M; Salzmann, Gian M; Schmal, Hagen;

    2012-01-01

    BACKGROUND: Autologous chondrocyte implantation (ACI) is a well-established treatment option for isolated cartilage defects of the knee joint, providing satisfying outcome. However, cases of treatment failure with the need for surgical reintervention are reported; typical patient's individual and...

  14. Exploration of mechanisms underlying the strain-rate-dependent mechanical property of single chondrocytes

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    Nguyen, Trung Dung; Gu, YuanTong, E-mail: yuantong.gu@qut.edu.au [School of Chemistry, Physics and Mechanical Engineering, Queensland University of Technology, Brisbane, Queensland (Australia)

    2014-05-05

    Based on the characterization by Atomic Force Microscopy, we report that the mechanical property of single chondrocytes has dependency on the strain-rates. By comparing the mechanical deformation responses and the Young's moduli of living and fixed chondrocytes at four different strain-rates, we explore the deformation mechanisms underlying this dependency property. We found that the strain-rate-dependent mechanical property of living cells is governed by both of the cellular cytoskeleton and the intracellular fluid when the fixed chondrocytes are mainly governed by their intracellular fluid, which is called the consolidation-dependent deformation behavior. Finally, we report that the porohyperelastic constitutive material model which can capture the consolidation-dependent behavior of both living and fixed chondrocytes is a potential candidature to study living cell biomechanics.

  15. The cytoskeleton of chondrocytes of Sepia officinalis (Mollusca, Cephalopoda: an immunocytochemical study

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    F Leone

    2009-06-01

    Full Text Available Our previous electron microscope study showed that chondrocytes from cephalopod cartilage possess a highly developed cytoskeleton and numerous cytoplasmic processes that ramify extensively through the tissue. We have now carried out a light microscope immunocytochemical study of chondrocytes from the orbital cartilage of Sepia officinalis to obtain indications as to the nature of the cytoskeletal components. We found clear positivity to antibodies against mammalian tubulin, vimentin, GFAP, and actin, but not keratin. The simultaneous presence of several cytoskeletal components is consistent with the hypothesis that cephalopod chondrocytes have the characteristics of both chondrocytes and osteocytes of vertebrates, which endow the tissue as a whole with some of the properties of vertebrate bone. We confirm, therefore, the presence in molluscs of the ubiquitous cytoskeletal proteins of metazoan cells that have remained highly conserved throughout phylogenetic evolution.

  16. Mechanical overloading causes mitochondrial superoxide and SOD2 imbalance in chondrocytes resulting in cartilage degeneration.

    Science.gov (United States)

    Koike, Masato; Nojiri, Hidetoshi; Ozawa, Yusuke; Watanabe, Kenji; Muramatsu, Yuta; Kaneko, Haruka; Morikawa, Daichi; Kobayashi, Keiji; Saita, Yoshitomo; Sasho, Takahisa; Shirasawa, Takuji; Yokote, Koutaro; Kaneko, Kazuo; Shimizu, Takahiko

    2015-06-25

    Mechanical stress and aging are major risk factors of cartilage degeneration. Human studies have previously reported that oxidative damage increased, while SOD2 protein was reciprocally downregulated in osteoarthritic degenerated cartilage. However, it remains unclear whether mitochondrial superoxide imbalance in chondrocytes causes cartilage degeneration. We herein demonstrate that mechanical loading promoted mitochondrial superoxide generation and selective Sod2 downregulation in chondrocytes in vivo and that mitochondrial superoxide inducer also downregulated Sod2 expression in chondrocytes in vitro. A genetically manipulated model revealed that Sod2 deficiency in chondrocytes also resulted in mitochondrial superoxide overproduction and dysfunction, thus leading to cartilage degeneration. Intra-articular injection of a permeable antioxidant effectively suppressed the mechanical loading-induced mitochondrial superoxide generation and cartilage degeneration in mice. Our findings demonstrate that mitochondrial superoxide plays a pivotal role in the development and progression of osteoarthritis, and the mitochondrial superoxide balance may therefore be a promising target for the treatment of cartilage degeneration.

  17. Role of Insulin-Transferrin-Selenium in Auricular Chondrocyte Proliferation and Engineered Cartilage Formation in Vitro

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    Xia Liu

    2014-01-01

    Full Text Available The goal of this study is to determine the effects of Insulin-Transferrin-Selenium (ITS on proliferation of auricular chondrocytes and formation of engineered cartilage in vitro. Pig auricular monolayer chondrocytes and chondrocyte pellets were cultured in media containing 1% ITS at different concentrations of fetal bovine serum (FBS, 10%, 6%, 2%, 0%, or 10% FBS alone as a control for four weeks. Parameters including cell proliferation in monolayer, wet weight, collagen type I/II/X (Col I, II, X and glycosaminoglycan (GAG expression, GAG content of pellets and gene expression associated with cartilage formation/dedifferentiation (lost cartilage phenotype/hypertrophy within the chondrocyte pellets were assessed. The results showed that chondrocytes proliferation rates increased when FBS concentrations increased (2%, 6%, 10% FBS in ITS supplemented groups. In addition, 1% ITS plus 10% FBS significantly promoted cell proliferation than 10% FBS alone. No chondrocytes grew in ITS alone medium. 1% ITS plus 10% FBS enhanced cartilage formation in terms of size, wet weight, cartilage specific matrices, and homogeneity, compared to 10% FBS alone group. Furthermore, ITS prevented engineered cartilage from dedifferentiation (i.e., higher index of Col II/Col I mRNA expression and expression of aggrecan and hypertrophy (i.e., lower mRNA expression of Col X and MMP13. In conclusion, our results indicated that ITS efficiently enhanced auricular chondrocytes proliferation, retained chondrogenic phenotypes, and promoted engineered cartilage formation when combined with FBS, which is potentially used as key supplementation in auricular chondrocytes and engineered cartilage culture.

  18. Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

    Science.gov (United States)

    Gradišnik, Lidija; Gorenjak, Mario; Vogrin, Matjaž

    2017-01-01

    Background Cartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue. Methods Isolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA) was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2), collagen 1 (COL1) and aggrecan (ACAN) was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production. Results Cartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well. Discussion In this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a common and very

  19. Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA

    Directory of Open Access Journals (Sweden)

    Jakob Naranda

    2017-03-01

    Full Text Available Background Cartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue. Methods Isolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2, collagen 1 (COL1 and aggrecan (ACAN was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production. Results Cartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well. Discussion In this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a

  20. Human chondrocytes respond discordantly to the protein encoded by the osteoarthritis susceptibility gene GDF5.

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    Madhushika Ratnayake

    Full Text Available A genetic deficit mediated by SNP rs143383 that leads to reduced expression of GDF5 is strongly associated with large-joint osteoarthritis. We speculated that this deficit could be attenuated by the application of exogenous GDF5 protein and as a first step we have assessed what effect such application has on primary osteoarthritis chondrocyte gene expression. Chondrocytes harvested from cartilage of osteoarthritic patients who had undergone joint replacement were cultured with wildtype recombinant mouse and human GDF5 protein. We also studied variants of GDF5, one that has a higher affinity for the receptor BMPR-IA and one that is insensitive to the GDF5 antagonist noggin. As a positive control, chondrocytes were treated with TGF-β1. Chondrocytes were cultured in monolayer and micromass and the expression of genes coding for catabolic and anabolic proteins of cartilage were measured by quantitative PCR. The expression of the GDF5 receptor genes and the presence of their protein products was confirmed and the ability of GDF5 signal to translocate to the nucleus was demonstrated by the activation of a luciferase reporter construct. The capacity of GDF5 to elicit an intracellular signal in chondrocytes was demonstrated by the phosphorylation of intracellular Smads. Chondrocytes cultured with TGF-β1 demonstrated a consistent down regulation of MMP1, MMP13 and a consistent upregulation of TIMP1 and COL2A1 with both culture techniques. In contrast, chondrocytes cultured with wildtype GDF5, or its variants, did not show any consistent response, irrespective of the culture technique used. Our results show that osteoarthritis chondrocytes do not respond in a predictable manner to culture with exogenous GDF5. This may be a cause or a consequence of the osteoarthritis disease process and will need to be surmounted if treatment with exogenous GDF5 is to be advanced as a potential means to overcome the genetic deficit conferring osteoarthritis

  1. Canine chondrocytes seeded in type I and type II collagen implants investigated in vitro.

    Science.gov (United States)

    Nehrer, S; Breinan, H A; Ramappa, A; Shortkroff, S; Young, G; Minas, T; Sledge, C B; Yannas, I V; Spector, M

    1997-01-01

    Synthetic and natural absorbable polymers have been used as vehicles for implantation of cells into cartilage defects to promote regeneration of the articular joint surface. Implants should provide a pore structure that allows cell adhesion and growth, and not provoke inflammation or toxicity when implanted in vivo. The scaffold should be absorbable and the degradation should match the rate of tissue regeneration. To facilitate cartilage repair the chemical structure and pore architecture of the matrix should allow the seeded cells to maintain the chondrocytic phenotype, characterized by synthesis of cartilage-specific proteins. We investigated the behavior of canine chondrocytes in two spongelike matrices in vitro: a collagen-glycosaminoglycan (GAG) copolymer produced from bovine hide consisting of type I collagen and a porous scaffold made of type II collagen by extraction of porcine cartilage. Canine chondrocytes were seeded on both types of matrices and cultured for 3 h, 7 days, and 14 days. The histology of chondrocyte-seeded implants showed a significantly higher percentage of cells with spherical morphology, consistent with chondrocytic morphology, in the type II sponge at each time point. Pericellular matrix stained for proteoglycans and for type II collagen after 14 days. Biochemical analysis of the cell seeded sponges for GAG and DNA content showed increases with time. At day 14 there was a significantly higher amount of DNA and GAG in the type II matrix. This is the first study that directly compares the behavior of chondrocytes in type I and type II collagen matrices. The type II matrix may be of value as a vehicle for chondrocyte implantation on the basis of the higher percentage of chondrocytes retaining spherical morphology and greater biosynthetic activity that was reflected in the greater increase of GAG content.

  2. Effect of microcavitary alginate hydrogel with different pore sizes on chondrocyte culture for cartilage tissue engineering

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    Zeng, Lei; Yao, Yongchang [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China); Wang, Dong-an, E-mail: DAWang@ntu.edu.sg [National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China); Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore 637457 (Singapore); Chen, Xiaofeng, E-mail: chenxf@scut.edu.cn [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China)

    2014-01-01

    In our previous work, a novel microcavitary hydrogel was proven to be effective for proliferation of chondrocytes and maintenance of chondrocytic phenotype. In present work, we further investigated whether the size of microcavity would affect the growth and the function of chondrocytes. By changing the stirring rate, gelatin microspheres in different sizes including small size (80–120 μm), middle size (150–200 μm) and large size (250–300 μm) were prepared. And then porcine chondrocytes were encapsulated into alginate hydrogel with various sizes of gelatin microspheres. Cell Counting Kit-8 (CCK-8), Live/dead staining and real-time PCR were used to analyze the effect of the pore size on cell proliferation and expression of specific chondrocytic genes. According to all the data, cells cultivated in microcavitary hydrogel, especially in small size, had preferable abilities of proliferation and higher expression of cartilaginous markers including type II collagen, aggrecan and cartilage oligomeric matrix protein (COMP). Furthermore, it was shown by western blot assay that the culture of chondrocytes in microcavitary hydrogel could improve the proliferation of cells potentially by inducing the Erk1/2-MAPK pathway. Taken together, this study demonstrated that chondrocytes favored microcavitary alginate hydrogel with pore size within the range of 80–120 μm for better growth and ECM synthesis, in which Erk1/2 pathway was involved. This culture system would be promising for cartilage tissue engineering. - Highlights: • A novel model with microcavitary structure was set up to study the interaction between cells and materials. • Microcavitary alginate hydrogel could enhance the proliferation of chondrocytes and promote the expression of cartilaginous genes as compared with plain alginate hydrogel. • Cells in microcavitary alginate hydrogel with pore size within the range of 80–120 μm were capable of better growth and ECM synthesis.

  3. Long Term Effect on Professionals' Knowledge, Practice and Attitudes towards User Involvement Four Years after Implementing an Organisational Development Plan: A Controlled Study.

    Directory of Open Access Journals (Sweden)

    Marit By Rise

    Full Text Available Health service organisations are increasingly implementing user involvement initiatives according to requirements from governments, such as user representation in administrational boards, better information to users, and more involvement of the users during treatment. Professionals are vital in all initiatives to enhance user involvement, and initiatives to increase involvement should influence the professionals' practice and attitudes. The implementation of a development plan intending to enhance user involvement in a mental health hospital in Central Norway had no effect on the professionals after 16 months. The objective was therefore to investigate the long term effect on the professionals' knowledge, practice and attitudes towards user involvement after four years.This was a non-randomized controlled study including professionals from three mental health hospitals in Central Norway. A development plan intended to enhance user participation was implemented in one of the hospitals, including establishing a patient education centre and a user office, purchasing of user expertise, appointing contact professionals for next of kin, and improving of the centre's information and the professional culture. The professionals at two other hospitals constituted the control group. All professionals were invited to answer the Consumer Participation Questionnaire (CPQ and additional questions, at a four year interval.A total of 399 professionals participated (43% response rate. Comparing the changes in the intervention group with the changes in the control group, the results showed that the plan had improved some aspects of the professionals' knowledge about the user involvement taking place in the hospital. In addition, some parts of the professionals' practice of providing information to the service users was improved, and the development plan might have raised their awareness about insufficient involvement of next of kin.This is the first controlled

  4. Physical Activity and Fruit and Vegetable Intake among Black and White Female College Students at Two- and Four-Year Colleges and Universities

    Science.gov (United States)

    Goodwin, Sherell Brown; Stratton, Erin; Lowe, Kincaid; Grimsley, Linda; Rodd, Jan; Williams, Catherine; Mattox, Cheri; Foster, Bruce

    2014-01-01

    Objective We compared fruit and vegetable (FV) intake, physical activity (PA), and overweight/ obesity among Black and White females attending two- and four-year colleges. Methods We recruited 24,055 students at six colleges in the Southeast to complete an online survey, yielding 4840 responses (20.1% response rate). The current analyses focused on the 2276 Black and White females. Results Binary logistic regression analyses indicated that recommended FV intake among White females was associated with greater extraversion (Odds Ratio [OR] = 1.05, 95% Confidence Interval [CI] 1.00, 1.11, p = 0.05) and greater conscientiousness (OR = 1.08, CI 1.02, 1.14, p = 0.01), whereas among Black females correlates included greater openness to experience (OR = 1.08, CI 1.01, 1.15, p = 0.03). Ordinal logistic regression analyses indicated that, among White females, greater PA was associated with attending a four-year college (Beta = 0.27, CI 0.01, 0.53, p = 0.04), whereas among Black females, correlates were with younger age (Beta = 0.01, CI 0.17, 0.03, p = 0.003) and greater emotional stability (Beta = 0.07, CI 0.01, 0.13, p = 0.02). Binary logistic regression analyses indicated that, among White females, being overweight or obese was associated with older age (OR = 1.08, CI 1.01, 1.16, p = 0.03), attending a two-year college (OR = 1.62, CI 1.22, 2.16, p = 0.001), and lower satisfaction with life (OR = 0.96, CI 0.94, 0.98, p = 0.002), whereas among Black females, being overweight or obese was associated with older age (OR = 1.87, CI 1.10, 1.28, p < 0.001). Conclusions Identifying factors related to obesity-related factors is critical in developing interventions targeting factors associated with overweight and obesity among Black and White females attending two- and four-year colleges. Moreover, understanding different college settings and the contextual factors associated with overweight/obesity is critical. PMID:25328836

  5. In vitro isolation and cultivation of human chondrocytes for osteoarthritis renovation.

    Science.gov (United States)

    Xu, Jiaming; Zhang, Changqing

    2014-08-01

    The purpose of this study was to evaluate the repair effects of chondrocytes that were cultured in vitro on osteoarthritis (OA). Chondrocytes were isolated from fetal rabbits and cultured in Biosilon microcarriers. Sixty rabbits were randomly divided into three groups equally (blank group, model group, treatment group). The rabbit knee OA model was established by inducing papain. Rabbits in the treatment group were injected with the chondrocytes that were cultured in vitro. Hematoxylin-eosin (HE) staining and gross morphologic observation were conducted. Expression level of cytokines such as IL-1bβ, IL-6, and TNF-α in cartilage synovial cells was also analyzed by an ELISA assay. The cultured chondrocyte was validated by a positive stain of type II collagen and vimentin by immunofluorescence. Compared to the model group, the articular cartilage of the rabbit knee in the treatment group showed a normal color, smooth surface, and none of malacia and coloboma. HE staining indicated that the articular surface of the treatment group tended to be smooth and flat; the matrix stained tinge and the cartilage destruction and fiber hyperplasia of the synovia were lightened. The expression levels of IL-1bβ, IL-6, and TNF-α also declined in the treatment group. OA symptoms were improved by treating with chondrocytes. In summary, the animal experiment in the present study indicated that chondrocyte injection played an active effect on renovation of OA.

  6. Characterization of pediatric microtia cartilage: a reservoir of chondrocytes for auricular reconstruction using tissue engineering strategies.

    Science.gov (United States)

    Melgarejo-Ramírez, Y; Sánchez-Sánchez, R; García-López, J; Brena-Molina, A M; Gutiérrez-Gómez, C; Ibarra, C; Velasquillo, C

    2016-09-01

    The external ear is composed of elastic cartilage. Microtia is a congenital malformation of the external ear that involves a small reduction in size or a complete absence. The aim of tissue engineering is to regenerate tissues and organs clinically implantable based on the utilization of cells and biomaterials. Remnants from microtia represent a source of cells for auricular reconstruction using tissue engineering. To examine the macromolecular architecture of microtia cartilage and behavior of chondrocytes, in order to enrich the knowledge of this type of cartilage as a cell reservoir. Auricular cartilage remnants were obtained from pediatric patients with microtia undergoing reconstructive procedures. Extracellular matrix composition was characterized using immunofluorescence and histological staining methods. Chondrocytes were isolated and expanded in vitro using a mechanical-enzymatic protocol. Chondrocyte phenotype was analyzed using qualitative PCR. Microtia cartilage preserves structural organization similar to healthy elastic cartilage. Extracellular matrix is composed of typical cartilage proteins such as type II collagen, elastin and proteoglycans. Chondrocytes displayed morphological features similar to chondrocytes derived from healthy cartilage, expressing SOX9, COL2 and ELN, thus preserving chondral phenotype. Cell viability was 94.6 % during in vitro expansion. Elastic cartilage from microtia has similar characteristics, both architectural and biochemical to healthy cartilage. We confirmed the suitability of microtia remnant as a reservoir of chondrocytes with potential to be expanded in vitro, maintaining phenotypical features and viability. Microtia remnants are an accessible source of autologous cells for auricular reconstruction using tissue engineering strategies.

  7. Efficiency of Human Epiphyseal Chondrocytes with Differential Replication Numbers for Cellular Therapy Products

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    Michiyo Nasu

    2016-01-01

    Full Text Available The cell-based therapy for cartilage or bone requires a large number of cells; serial passages of chondrocytes are, therefore, needed. However, fates of expanded chondrocytes from extra fingers remain unclarified. The chondrocytes from human epiphyses morphologically changed from small polygonal cells to bipolar elongated spindle cells and to large polygonal cells with degeneration at early passages. Gene of type II collagen was expressed in the cells only at a primary culture (Passage 0 and Passage 1 (P1 cells. The nodules by implantation of P0 to P8 cells were composed of cartilage and perichondrium. The cartilage consisted of chondrocytes with round nuclei and type II collagen-positive matrix, and the perichondrium consisted of spindle cells with type I collage-positive matrix. The cartilage and perichondrium developed to bone with marrow cavity through enchondral ossification. Chondrogenesis and osteogenesis by epiphyseal chondrocytes depended on replication number in culture. It is noteworthy to take population doubling level in correlation with pharmaceutical efficacy into consideration when we use chondrocytes for cell-based therapies.

  8. Human pituitary tissue secretes a potent growth factor for chondrocyte proliferation.

    Science.gov (United States)

    Kasper, S; Friesen, H G

    1986-01-01

    We report the secretion from human pituitary tumor fragments in organ culture of a potent mitogen for chondrocyte proliferation. Primary human pituitary cell and organ cultures were established from pituitary fragments obtained from patients with acromegaly, prolactinomas, and nonfunctional adenomas. The conditioned culture medium contained a mitogenic factor(s) that stimulated rabbit fetal chondrocyte proliferation, causing up to an 8-fold increase in cell number when added to Ham's F-10 medium in the presence of 10% fetal bovine serum. Blood leaking into the surgical field after the adenomectomy is known to contain very high concentrations of pituitary hormones. Serum samples, obtained from this venous "ooze" collected at the site of pituitary surgery, also were found to contain chondrocyte growth-promoting activity. Some venous serum samples stimulated chondrocyte proliferation in a dose-dependent manner down to a 1:10 dilution of 1 microliter serum, indicating that the material being secreted was very potent indeed. Gel filtration on Sephadex G-100 and analytical gel isoelectric focusing of culture media or serum samples from the pituitary fossa demonstrated that the growth factor secreted from the pituitary tumor fragments as well as from the venous serum is similar, if not identical, to chondrocyte growth factor (mol wt, 43,000; pI 7.6-7.9) purified from human pituitaries collected at autopsy. These results suggest that the chondrocyte growth-promoting factor(s) may not only be secreted by pituitary tumor fragments but by normal human pituitary tissue as well.

  9. GROWTH DIFFERENTIATION FACTOR-5 STIMULATES THE GROWTH AND ANABOLIC METABOLISM OF ARTICULAR CHONDROCYTES

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Objective To observe the effect of growth differentiation factor-5 (GDF-5) on the growth and anabolic metabolism of articular chondrocytes. Methods The articular chondrocytes isolated from rats were treated with various concentrations of rmGDF-5, and the growth of chondrocytes measured by MTT assay, the cellular cartilage matrices formation detected sulfated glycosaminoglycan by Alcian blue staining and type Ⅱcollagen by RT-PCR. Results After 7 days culture, MTT assay showed that GDF-5 enhanced the growth of chondrocytes in a dose-dependent manner, RT-PCR showed that GDF-5 clearly induced the synthesis of type Ⅱ collagen because of the col2a1 mRNA band more and more strong in a dose-dependent. Chondrocytes were cultured with GDF-5 for 14 days, the intensity of Alcian blue staining was greatly enhanced, especially, at a high concentration of blue-stainable material in a concentration-dependent manner and in a does-dependent manner. Conclusion GDF-5 enhanced the growth of mature articular chondrocytes, and stimulated the cellular cartilage matrices formation in mono-layer culture.

  10. Finite difference time domain model of ultrasound propagation in agarose scaffold containing collagen or chondrocytes.

    Science.gov (United States)

    Inkinen, Satu I; Liukkonen, Jukka; Malo, Markus K H; Virén, Tuomas; Jurvelin, Jukka S; Töyräs, Juha

    2016-07-01

    Measurement of ultrasound backscattering is a promising diagnostic technique for arthroscopic evaluation of articular cartilage. However, contribution of collagen and chondrocytes on ultrasound backscattering and speed of sound in cartilage is not fully understood and is experimentally difficult to study. Agarose hydrogels have been used in tissue engineering applications of cartilage. Therefore, the aim of this study was to simulate the propagation of high frequency ultrasound (40 MHz) in agarose scaffolds with varying concentrations of chondrocytes (1 to 32 × 10(6) cells/ml) and collagen (1.56-200 mg/ml) using transversely isotropic two-dimensional finite difference time domain method (FDTD). Backscatter and speed of sound were evaluated from the simulated pulse-echo and through transmission measurements, respectively. Ultrasound backscatter increased with increasing collagen and chondrocyte concentrations. Furthermore, speed of sound increased with increasing collagen concentration. However, this was not observed with increasing chondrocyte concentrations. The present study suggests that the FDTD method may have some applicability in simulations of ultrasound scattering and propagation in constructs containing collagen and chondrocytes. Findings of this study indicate the significant role of collagen and chondrocytes as ultrasound scatterers and can aid in development of modeling approaches for understanding how cartilage architecture affects to the propagation of high frequency ultrasound.

  11. Central odontogenic fibroma (simple type) in a four year old boy: Atypical cone-beam computed tomographic appearance with periosteal reaction

    Energy Technology Data Exchange (ETDEWEB)

    Anbiaee, Najme; Ebrahimnejad, Hamed; Sanaei, Alireza [Dept. of , Oral and Maxillofacial Radiology, Maxillofacial Diseases Research Center, School of Dentistry, Mashhad University of Medical Sciences, Mashhad (Iran, Islamic Republic of)

    2015-06-15

    Central odontogenic fibroma (COF) is a rare benign tumor that accounts for 0.1% of all odontogenic tumors. A case of COF (simple type) of the mandible in a four-year-old boy is described in this report. The patient showed asymptomatic swelling in the right inferior border of the lower jaw for one week. A panoramic radiograph showed a poorly-defined destructive unilocular radiolucent area. Cone-beam computed tomography showed expansion and perforation of the adjacent cortical bone plates. A periosteal reaction with the Codman triangle pattern was clearly visible in the buccal cortex. Since the tumor had destroyed a considerable amount of bone, surgical resection was performed. No recurrence was noted.

  12. Compliance in the First Four Years of AAS 27 Financial Reporting by Local Government: A Study of the New South Wales Experience

    Directory of Open Access Journals (Sweden)

    Gregory Kenneth Laing

    2007-06-01

    Full Text Available This paper reports on a longitudinal study of the levels of compliance for the first fouryears (1994 to1997 following the introduction of the Australian Accounting Standard(AAS 27. The annual reports of all 177 local government councils in NSW wereanalysed revealing a high level of compliance with the standard. The main cause fornon-compliance was the complexity of the recognition and treatment of assets. Inaddition a number of problems were identified, in particular the timeliness of thereports and the disproportionately high levels of depreciation expenses reported in theoperating statement. The increased complexities of the financial management in LocalGovernment imposed by AAS 27 required professional accounting competencies moreappropriately derived from a degree level education which contributed to a highturnover of accounting staff during the four years. Finally, the paper questions theviability of the proposed dismantling of AAS 27 and the possible impact this might haveupon the sector.

  13. Type II collagen peptide is able to accelerate embryonic chondrocyte differentiation: an association with articular cartilage matrix resorption in osteoarthrosis

    Directory of Open Access Journals (Sweden)

    Elena Vasil'evna Chetina

    2010-01-01

    Conclusion. The effect of CP on gene expression and collagen decomposition activity depends on the morphotype of embryonic chondrocytes. Lack of effect of CP on collagen decomposition activity in both the embryonic hypertrophic chondrocytes and the cartilage explants from OA patients supports the hypothesis that the hypertrophic morphotype is a dominant morphotype of articular chondrocytes in OA. Moreover, collagen decomposition products can be involved in the resorption of matrix in OA and in the maintenance of chronic nature of the pathology.

  14. Normal age-related viscoelastic properties of chondrons and chondrocytes isolated from rabbit knee

    Institute of Scientific and Technical Information of China (English)

    DUAN Wang-ping; SUN Zhen-wei; LI Qi; LI Chun-jiang; WANG Li; CHEN Wei-yi; Jennifer Tickner; ZHENG Ming-hao; WEI Xiao-chun

    2012-01-01

    Background The mechanical microenvironment of the chondrocytes plays an important role in cartilage homeostasis and in the health of the joint.The pericellular matrix,cellular membrane of the chondrocytes,and their cytoskeletal structures are key elements in the mechanical environment.The aims of this study are to measure the viscoelastic properties of isolated chondrons and chondrocytes from rabbit knee cartilage using micropipette aspiration and to determine the effect of aging on these properties.Methods Three age groups of rabbit knees were evaluated:(1) young (2 months,n=10);(2) adult (8 months,n=10);and (3) old (31 months,n=10).Chondrocytes were isolated from the right knee cartilage and chondrons were isolated from left knees using enzymatic methods.Micropipette aspiration combined with a standard linear viscoelastic solid model was used to quantify changes in the viscoelastic properties of chondrons and chondrocytes within 2 hours of isolation.The morphology and structure of isolated chondrons were evaluated by optical microscope using hematoxylin and eosin staining and collagen-6 immunofluorescence staining.Results In response to an applied constant 0.3-0.4 kPa of negative pressure,all chondrocytes exhibited standard linear viscoelastic solid properties.Model predictions of the creep data showed that the average equilibrium modulus (E∞),instantaneous modulus (E0).and apparent viscosity (μ) of old chondrocytes was significantly lower than the young and adult chondrocytes (P<0.001);however,no difference was found between young and adult chondrocytes (P>0.05).The adult and old chondrons generally possessed a thicker pericellular matrix (PCM) with more enclosed cells.The young and adult chondrons exhibited the same viscoelastic creep behavior under a greater applied pressure (1.0-1.1kPa) without the deformation seen in the old chondrons.The viscoelastic properties (E∞,E0,and u) of young and adult chondrons were significantly greater than that observed

  15. [Grafting of carotid arteries].

    Science.gov (United States)

    Belov, Iu V; Stepanenko, A B; Gens, A P; Bazylev, V V; Seleznev, M N; Savichev, D D

    2005-01-01

    Over 5-years, 167 reconstructive surgeries for stenosis of internal carotid arteries (ICA) were performed in 124 patients. Mean age of the patients was 63.5 years. One hundred and twenty-nine carotid endarterectomies (CEAE) in 86 patients and 38 reconstructive operations of ICA in 38 patients were performed. There were no lethal outcomes in short- and long-term postoperative period. In short-term period after prosthesis of ICA restenosis was revealed in 3% patients, after eversion CEAE in 3% patients the embolism was seen, after standard CEAE restenosis were diagnosed in 8% patients and thrombosis -- in 3%. In long-term period after grafting of ICA the strokes were seen in 3%, stenosis -- in 6% patients, after eversion endarterectomy -- in 0 and 3% patients, and after standard CEAE -- in 3 and 24% patients, respectively. It is concluded that grafting of ICA is adequate surgical method of reconstruction and stroke prevention in specific variants of carotid atherosclerosis.

  16. Influence of Material Properties on Rate of Resorption of Two Bone Graft Materials after Sinus Lift Using Radiographic Assessment

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    Fawzi Riachi

    2012-01-01

    Full Text Available Purpose. The aim of this study was to investigate the influence of chemical and physical properties of two graft materials on the rate of resorption. Materials and Methods. Direct sinus graft procedure was performed on 22 patients intended for implant placement. Two types of graft materials were used (Bio-Oss and Cerabone and after 8 months healing time the implants were inserted. Radiographic assessment was performed over the period of four years. Particle size, rate of calcium release, and size and type of crystal structure of each graft were evaluated. Results. The average particle size of Bio-Oss (1 mm was much smaller compared to Cerabone (2.7 mm. The amount of calcium release due to dissolution of material in water was much higher for Bio-oss compared to Cerabone. X-ray image analysis revealed that Bio-Oss demonstrated significantly higher volumetric loss (33.4 ± 3.1% of initial graft size compared to Cerabone (23.4 ± 3.6%. The greatest amount of vertical loss of graft material volume was observed after one year of surgery. Conclusion. The chemical and physical properties of bone graft material significantly influence resorption rate of bone graft materials used for sinus augmentation.

  17. In vitro and in vivo validation of human and goat chondrocyte labeling by green fluorescent protein lentivirus transduction.

    Science.gov (United States)

    Miot, Sylvie; Gianni-Barrera, Roberto; Pelttari, Karoliina; Acharya, Chitrangada; Mainil-Varlet, Pierre; Juelke, Henriette; Jaquiery, Claude; Candrian, Christian; Barbero, Andrea; Martin, Ivan

    2010-02-01

    We investigated whether human articular chondrocytes can be labeled efficiently and for long-term with a green fluorescent protein (GFP) lentivirus and whether the viral transduction would influence cell proliferation and tissue-forming capacity. The method was then applied to track goat articular chondrocytes after autologous implantation in cartilage defects. Expression of GFP in transduced chondrocytes was detected cytofluorimetrically and immunohistochemically. Chondrogenic capacity of chondrocytes was assessed by Safranin-O staining, immunostaining for type II collagen, and glycosaminoglycan content. Human articular chondrocytes were efficiently transduced with GFP lentivirus (73.4 +/- 0.5% at passage 1) and maintained the expression of GFP up to 22 weeks of in vitro culture after transduction. Upon implantation in nude mice, 12 weeks after transduction, the percentage of labeled cells (73.6 +/- 3.3%) was similar to the initial one. Importantly, viral transduction of chondrocytes did not affect the cell proliferation rate, chondrogenic differentiation, or tissue-forming capacity, either in vitro or in vivo. Goat articular chondrocytes were also efficiently transduced with GFP lentivirus (78.3 +/- 3.2%) and maintained the expression of GFP in the reparative tissue after orthotopic implantation. This study demonstrates the feasibility of efficient and relatively long-term labeling of human chondrocytes for co-culture on integration studies, and indicates the potential of this stable labeling technique for tracking animal chondrocytes for in cartilage repair studies.

  18. Expression of caspase-3 and -9 relevant to cartilage destruction and chondrocyte apoptosis in human osteoarthritic cartilage.

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    Matsuo M

    2001-12-01

    Full Text Available To clarify the involvement of the caspase family in the pathway of NO-induced chondrocyte apoptosis, osteoarthritis (OA cartilage obtained from 8 patients undergoing total hip arthroplasty were used for histopathological study. Cartilage samples taken from non-fibrillated areas of femoral head resected during surgery for femoral neck fracture were used for comparison. DNA fragmentation of chondrocytes was detected by the nick end-labeling (TUNEL method. Apoptosis was further confirmed by transmission electron microscopy. The distributions of nitrotyrosine (NT, caspase-3, and -9 were examined immunohistochemically. The populations of apoptotic as well as NT-, caspase-3-, and -9-positive cells were quantified by counting the number of cells in the superficial, middle, and deep layers, respectively. The TUNEL-positive cells were observed primarily in superficial proliferating chondrocytes, clustering chondrocytes, and deep-layer chondrocytes of OA cartilage. Few positive cells were seen in the proliferating chondrocytes in the middle layer. Positive reactions for caspase-3 and -9 were observed in chondrocytes in similar areas. Histological OA grade showed significant correlations with the mean populations of apoptotic chondrocytes (% apoptosis over the 3 areas. The populations of NT-positive cells (% NT over the same areas also showed significant correlation with OA grade. Positivity for caspase-3 closely correlated with the OA grade, % apoptosis and %NT. It was concluded that caspase-3 and -9 could play a role in NO-induced chondrocyte apoptosis in OA cartilage.

  19. Buccal Mucosal Graft Urethroplasty

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    Angela M. Arlen

    2010-01-01

    Full Text Available At our institution, the majority of buccal mucosal graft urethroplasties are performed using a two-team approach with an otolaryngologic surgeon. We report our two-surgeon experience with buccal mucosal grafting for reconstruction of all anterior urethral strictures. Twenty-four men underwent autologous buccal mucosal graft urethroplasty between October 2001 and September 2008 for recurrent urethral stricture disease. Twenty-two underwent a single-stage repair and two underwent a two-stage repair. Medical charts were retrospectively reviewed for demographics, comorbidities, etiology, location and length of stricture, and prior interventions in order to identify predictors of buccal urethroplasty success, defined as no evidence of stricture recurrence. All patients underwent retrograde urethrogram and cystoscopy. Operative and anesthesia times were evaluated. We determined an overall success rate of 83.3% (20 of 24 cases. Mean anesthesia time for single-stage urethroplasty was 155 min and mean operative time was 123 min. One of the two two-stage urethroplasties experienced stricture recurrence (50%. The single-stage buccal graft success rate was 86.4% (19 of 22 cases. Two of the four who developed recurrent stricture disease that required intervention had undergone a previous mesh urethroplasty. Complications developed in four of 24 patients (16.6%, including superficial wound infection (one, superficial wound dehiscence (two, and abscess/fistula formation requiring reoperation (one. The buccal mucosa is an ideal tissue for both single- and two-stage substitution urethroplasty for patients with recurrent stricture disease. Our two-surgeon technique minimizes anesthesia and operative times, and contributes to the overall high success rate and relatively low complication rate.

  20. Intermediate- to Long-Term Results of Combined Anterior Cruciate Ligament Reconstruction and Autologous Chondrocyte Implantation

    Science.gov (United States)

    Pike, Andrew N.; Bryant, Tim; Ogura, Takahiro; Minas, Tom

    2017-01-01

    Background: Cartilage injury associated with anterior cruciate ligament (ACL) ruptures is common; however, relatively few reports exist on concurrent cartilage repair with ACL reconstruction. Autologous chondrocyte implantation (ACI) has been utilized successfully for treatment of moderate to large chondral defects. Hypothesis: ACL insufficiency with relatively large chondral defects may be effectively managed with concurrent ACL reconstruction and ACI. Study Design: Case series; Level of evidence, 4. Methods: Patients undergoing concurrent ACL primary or revision reconstruction with ACI of single or multiple cartilage defects were prospectively evaluated for a minimum 2 years. Pre- and postoperative outcome measures included the modified Cincinnati Rating Scale (MCRS), Western Ontario and McMaster Universities Osteoarthritis Index, visual analog pain scales, and postsurgery satisfaction surveys. ACI graft failure or persistent pain without functional improvement were considered treatment failures. Results: Twenty-six patients were included, with 13 primary and 13 revision ACL reconstructions performed. Mean defect total surface area was 8.4 cm2, with a mean follow-up of 95 months (range, 24-240 months). MCRS improved from 3.62 ± 1.42 to 5.54 ± 2.32, Western Ontario and McMaster Universities Osteoarthritis Index from 45.31 ± 17.27 to 26.54 ± 17.71, and visual analog pain scale from 6.19 ± 1.27 to 3.65 ± 1.77 (all Ps <.001). Eight patients were clinical failures, 69% of patients were improved at final follow-up, and 92% stated they would likely undergo the procedure again. No outcome correlation was found with regard to age, body mass index, sex, defect size/number, follow-up time, or primary versus revision ACL reconstruction. In subanalysis, revision ACL reconstructions had worse preoperative MCRS scores and greater defect surface areas. However, revision MCRS score improvements were greater, resulting in similar final functional scores when compared with

  1. A novel fibrin gel derived from hyaluronic acid-grafted fibrinogen

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    Yang, Chiung L; Chen, Hui W; Wang, Tzu C; Wang, Yng J, E-mail: wang@ym.edu.tw [Institute of Biomedical Engineering, National Yang Ming University, No. 155, Sec. 2, Li-Nung St., Shih-Pai, Taipei, Taiwan 112 (China)

    2011-04-15

    Fibrinogen is a major plasma protein that forms a three-dimensional fibrin gel upon being activated by thrombin. In this study, we report the synthesis and potential applications of hybrid molecules composed of fibrinogen coupled to the reducing ends of short-chain hyaluronic acids (sHAs) by reductive amination. The grafting of sHAs to fibrinogen was verified by analyzing particle size, zeta potential and gel-electrophoretic mobility of the hybrid molecules. The sHA-fibrinogen hybrid molecules with graft ratios (sHA/fibrinogen) of up to 6.5 retained the ability to form gels in response to thrombin activation. The sHA-fibrin gels were transparent in appearance and exhibited high water content, which were characteristics distinct from those of gels formed by mixtures of sHAs and fibrinogen. The potential applications of the sHA-fibrin gels were evaluated. The sHA-fibrinogen gel with a graft ratio of 3.6 (S3.6F) was examined for its ability to encapsulate and support the differentiation of ATDC5 chondrocyte-like cells. Compared with the fibrinogen-formed gel, cells cultured in the S3.6F gel exhibited increased lacunae formation; moreover, the abundance of cartilaginous extracellular matrix molecules and the expression of chondrocyte marker genes, such as aggrecan, collagen II and Sox9, were also significantly increased. Our data suggest that the three-dimensional gel formed by the sHA-fibrinogen hybrid is a better support than the fibrin gel for chondrogenesis induction.

  2. Resveratrol protects rabbit articular chondrocyte against sodium nitroprusside-induced apoptosis via scavenging ROS.

    Science.gov (United States)

    Liang, Qian; Wang, Xiao-ping; Chen, Tong-sheng

    2014-09-01

    This study aims to investigate the mechanism by which resveratrol (RV) prevents sodium nitroprusside (SNP)-induced chondrocyte apoptosis, which is a characteristic feature of osteoarthritis (OA). Rabbit articular chondrocytes were pre-incubated with 100 μM RV for 18 h before 1.5 mM SNP co-treatment for 6 h. Cell viability was evaluated by CCK-8. Annexin V/PI double staining and Hoechst 33258 staining were used to determine the fashion of SNP-induced chondrocytes death. Mitochondrial membrane potential (ΔΨm) was measured by using flow cytometry (FCM) with TMRM and Rhodamine 123 staining. Intracellular reactive oxygen species (ROS) and nitric oxide (NO) levels were confirmed by FCM analysis with DCFH-DA and DAF-FM DA staining. Cytoskeleton proteins of chondrocytes co-stained with Actin-Trakcer Green and Tubulin-Trakcer Red were validated by confocal microscopy. SNP induced time- and dose-dependent chondrocytes apoptosis with decline of ΔΨm, activation of caspases as well as cytoskeletal remodeling. SNP induced a significant induction of both ROS and NO. RV remarkably prevented SNP-induced ROS production and apoptosis as well as cytoskeletal remodeling, but did not prevent SNP-induced NO production. Pretreatment with NO scavengers did not significantly prevent SNP-induced apoptosis and cytoskeletal remodeling. SNP induces NO-independent ROS production which dominates rabbit articular chondrocyte apoptosis, and RV protects chondrocytes against SNP-induced apoptosis via scavenging ROS instead of NO.

  3. Gold Nanoparticles of Diameter 13 nm Induce Apoptosis in Rabbit Articular Chondrocytes

    Science.gov (United States)

    Huang, Hao; Quan, Ying-yao; Wang, Xiao-ping; Chen, Tong-sheng

    2016-05-01

    Gold nanoparticles (AuNPs) have been widely used in biomedical science including antiarthritic agents, drug loading, and photothermal therapy. In this report, we studied the effects of AuNPs with diameters of 3, 13, and 45 nm, respectively, on rabbit articular chondrocytes. AuNPs were capped with citrate and their diameter and zeta potential were measured by dynamic light scattering (DLS). Cell viability was evaluated by Cell Counting Kit-8 (CCK-8) assay after the rabbit articular chondrocytes were pre-incubated with 3, 13, and 45 nm AuNPs, respectively, for 24 h. Flow cytometry (FCM) analysis with annexin V/propidium iodide (PI) double staining and fluorescence imaging with Hoechst 33258 staining were used to determine the fashion of AuNPs-induced chondrocyte death. Further, 13 nm AuNPs (2 nM) significantly induced chondrocyte death accompanying apoptotic characteristics including mitochondrial damage, externalization of phosphatidylserine and nuclear concentration. However, 3 nm AuNPs (2 nM) and 45 nm (0.02 nM) AuNPs did not induce cytotoxicity in chondrocytes. Although 13 nm AuNPs (2 nM) increased the intracellular reactive oxygen species (ROS) level, pretreatment with Nacetyl cysteine (NAC), a ROS scavenger, did not prevent the cytotoxicity induced by 13 nm AuNPs, indicating that 13 nm AuNPs (2 nM) induced ROS-independent apoptosis in chondrocytes. These results demonstrate the size-dependent cytotoxicity of AuNPs in chondrocytes, which must be seriously considered when using AuNPs for treatment of osteoarthritis (OA).

  4. Functional Characterization of TRPV4 As an Osmotically Sensitive Ion Channel in Articular Chondrocytes

    Science.gov (United States)

    Phan, Mimi N.; Leddy, Holly A.; Votta, Bartholomew J.; Kumar, Sanjay; Levy, Dana S.; Lipshutz, David B.; Lee, Sukhee; Liedtke, Wolfgang; Guilak, Farshid

    2010-01-01

    Objective Transient receptor potential vanilloid 4 (TRPV4) is a Ca2+ permeable channel that can be gated by tonicity (osmolarity) and mechanical stimuli. Chondrocytes, the cells in cartilage, respond to their osmotic and mechanical environments; however, the molecular basis of this signal transduction is not fully understood. The objective of this study was to demonstrate the presence and functionality of TRPV4 in chondrocytes. Methods TRPV4 protein expression was measured by immunolabeling and Western blotting. In response to TRPV4 agonist/antagonists, osmotic stress, and interleukin-1 (IL-1), changes in Ca2+ signaling, cell volume, and prostaglandin E2 (PGE2) production were measured in porcine chondrocytes using fluorescence microscopy, light microscopy, or immunoassay, respectively. Results TRPV4 was expressed abundantly at the RNA and protein level. Exposure to 4αPDD, a TRPV4 activator, caused Ca2+ signaling in chondrocytes, which was blocked by the selective TRPV4 antagonist, GSK205. Blocking TRPV4 diminished the chondrocytes' response to hypo-osmotic stress, reducing the fraction of Ca2+ responsive cells, regulatory volume decrease (RVD), and PGE2 production. Ca2+ signaling was inhibited by removal of extracellular Ca2+ or depletion of intracellular stores. Specific activation of TRPV4 restored defective RVD caused by IL-1. Chemical disruption of the primary cilium eliminated Ca2+ signaling in response to either 4αPDD or hypo-osmotic stress. Conclusion TRPV4 is present in articular chondrocytes, and chondrocyte response to hypo-osmotic stress is mediated by this channel, which involves both an extracellular Ca2+ and intracellular Ca2+ release. TRPV4 may also be involved in modulating the production or influence of pro-inflammatory molecules in response to osmotic stress. PMID:19790068

  5. Behaviour of oceanic 137Cs following the Fukushima Daiichi Nuclear Power Plant accident for four years simulated numerically by a regional ocean model

    Science.gov (United States)

    Tsumune, D.; Tsubono, T.; Aoyama, M.; Misumi, K.; Tateda, Y.

    2015-12-01

    A series of accidents at the Fukushima Dai-ichi Nuclear Power Plant (1F NPP) following the earthquake and tsunami of 11 March 2011 resulted in the release of radioactive materials to the ocean by two major pathways, direct release from the accident site and atmospheric deposition.We reconstructed spatiotemporal variability of 137Cs activity in the regional ocean for four years by numerical model, such as a regional scale and the North Pacific scale oceanic dispersion models, an atmospheric transport model, a sediment transport model, a dynamic biological compartment model for marine biota and river runoff model. Direct release rate of 137Cs were estimated for four years after the accident by comparing simulated results and observed activities very close to the site. The estimated total amounts of directly release was 3.6±0.7 PBq. Directly release rate of 137Cs decreased exponentially with time by the end of December 2012 and then, was almost constant. Decrease rate were quite small after 2013. The daily release rate of 137Cs was estimated to be the order of magnitude of 1010 Bq/day by the end of March 2015. The activity of directly released 137Cs was detectable only in the coastal zone after December 2012. Simulated 137Cs activities attributable to direct release were in good agreement with observed activities, a result that implies the estimated direct release rate was reasonable. There is no observed data of 137Cs activity in the ocean from 11 to 21 March 2011. Observed data of marine biota should reflect the history of 137Cs activity in this early period. We reconstructed the history of 137Cs activity in this early period by considering atmospheric deposition, river input, rain water runoff from the 1F NPP site. The comparisons between simulated 137Cs activity of marine biota by a dynamic biological compartment and observed data also suggest that simulated 137Cs activity attributable to atmospheric deposition was underestimated in this early period. The

  6. Assessing the fate of antibiotic contaminants in metal contaminated soils four years after cessation of long-term waste water irrigation

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    Tamtam, Fatima, E-mail: fatima.tamtam@upmc.fr [Laboratoire Hydrologie et Environnement, EPHE, UMR Sisyphe 7619, UPMC-Paris 6, 4 place Jussieu, BC 105, 75252 Paris Cedex (France); Ecole des Hautes Etudes en Sante Publique (EHESP), Laboratoire d' Etude et de Recherche en Environnement et Sante (LERES), Avenue Professeur Leon Bernard, Rennes (France); Oort, Folkert van [Inra, UR 251, Physicochimie et Ecotoxicologie des SolS d' Agrosystemes Contamines (Pessac), RD 10, F-78026 Versailles Cedex (France); Le Bot, Barbara [Ecole des Hautes Etudes en Sante Publique (EHESP), Laboratoire d' Etude et de Recherche en Environnement et Sante (LERES), Avenue Professeur Leon Bernard, Rennes (France); Dinh, Tuc [Laboratoire Hydrologie et Environnement, EPHE, UMR Sisyphe 7619, UPMC-Paris 6, 4 place Jussieu, BC 105, 75252 Paris Cedex (France); Mompelat, Sophie [Ecole des Hautes Etudes en Sante Publique (EHESP), Laboratoire d' Etude et de Recherche en Environnement et Sante (LERES), Avenue Professeur Leon Bernard, Rennes (France); Chevreuil, Marc [Laboratoire Hydrologie et Environnement, EPHE, UMR Sisyphe 7619, UPMC-Paris 6, 4 place Jussieu, BC 105, 75252 Paris Cedex (France); Lamy, Isabelle [Inra, UR 251, Physicochimie et Ecotoxicologie des SolS d' Agrosystemes Contamines (Pessac), RD 10, F-78026 Versailles Cedex (France); Thiry, Medard [MinesParisTech, Centre de Geoscience, ENSMP, UMR Sisyphe 7619, 35 Rue St Honore, 77305 Fontainebleau (France)

    2011-01-01

    Spreading of urban wastewater on agricultural land may lead to concomitant input of organic and inorganic pollutants. Such multiple pollution sites offer unique opportunities to study the fate of both heavy metals and pharmaceuticals. We examined the occurrence and fate of selected antibiotics in sandy-textured soils, sampled four years after cessation of 100 years irrigation with urban wastewater from the Paris agglomeration. Previous studies on heavy metal contamination of these soils guided our sampling strategy. Six antibiotics were studied, including quinolones, with a strong affinity for organic and mineral soil components, and sulfonamides, a group of more mobile molecules. Bulk samples were collected from surface horizons in different irrigation fields, but also in subsurface horizons in two selected profiles. In surface horizons, three quinolones (oxolinic acid, nalidixic acid, and flumequine) were present in eight samples out of nine. Their contents varied spatially, but were well-correlated one to another. Their distributions showed great similarities regarding spatial distribution of total organic carbon and heavy metal contents, consistent with a common origin by wastewater irrigation. Highest concentrations were observed for sampling sites close to irrigation water outlets, reaching 22 {mu}g kg{sup -1} for nalidixic acid. Within soil profiles, the two antibiotic groups demonstrated an opposite behavior: quinolones, found only in surface horizons; sulfamethoxazole, detected in clay-rich subsurface horizons, concomitant with Zn accumulation. Such distribution patterns are consistent with chemical adsorption properties of the two antibiotic groups: immobilization of quinolones in the surface horizons ascribed to strong affinity for organic matter (OM), migration of sulfamethoxazole due to a lower affinity for OM and its interception and retention in electronegative charged clay-rich horizons. Our work suggests that antibiotics may represent a durable

  7. Detecting new microRNAs in human osteoarthritic chondrocytes identifies miR-3085 as a human, chondrocyte-selective, microRNA

    Science.gov (United States)

    Crowe, N.; Swingler, T.E.; Le, L.T.T.; Barter, M.J.; Wheeler, G.; Pais, H.; Donell, S.T.; Young, D.A.; Dalmay, T.; Clark, I.M.

    2016-01-01

    Summary Objective To use deep sequencing to identify novel microRNAs (miRNAs) in human osteoarthritic cartilage which have a functional role in chondrocyte phenotype or function. Design A small RNA library was prepared from human osteoarthritic primary chondrocytes using in-house adaptors and analysed by Illumina sequencing. Novel candidate miRNAs were validated by northern blot and qRT-PCR. Expression was measured in cartilage models. Targets of novel candidates were identified by microarray and computational analysis, validated using 3′-UTR-luciferase reporter plasmids. Protein levels were assessed by western blot and functional analysis by cell adhesion. Results We identified 990 known miRNAs and 1621 potential novel miRNAs in human osteoarthritic chondrocytes, 60 of the latter were expressed in all samples assayed. MicroRNA-140-3p was the most highly expressed microRNA in osteoarthritic cartilage. Sixteen novel candidate miRNAs were analysed further, of which six remained after northern blot analysis. Three novel miRNAs were regulated across models of chondrogenesis, chondrocyte differentiation or cartilage injury. One sequence (novel #11), annotated in rodents as microRNA-3085-3p, was preferentially expressed in cartilage, dependent on chondrocyte differentiation and, in man, is located in an intron of the cartilage-expressed gene CRTAC-1. This microRNA was shown to target the ITGA5 gene directly (which encodes integrin alpha5) and inhibited adhesion to fibronectin (dependent on alpha5beta1 integrin). Conclusion Deep sequencing has uncovered many potential microRNA candidates expressed in human cartilage. At least three of these show potential functional interest in cartilage homeostasis and osteoarthritis (OA). Particularly, novel #11 (microRNA-3085-3p) which has been identified for the first time in man. PMID:26497608

  8. Aortic graft infection and mycotic aneurysm with Streptococcus equi zooepidemicus: two cases with favorable outcome of antibiotic treatment.

    Science.gov (United States)

    Altreuther, Martin; Lange, Conrad; Myhre, Hans Olav; Hannula, Raisa

    2013-02-01

    Infections with Streptococcus equi zooepidemicus are rare and are associated with contact with animals or animal products. There are very few reports about infected vascular grafts or aneurysms with this etiology. We present two patients. The first is a 77-year-old man with an infected bifurcated graft four years after an open operation for an abdominal aortic aneurysm (AAA). The second is a 72-year-old man with a symptomatic mycotic AAA, treated with endovascular aneurysm repair. Both received prolonged treatment with bactericidal antibiotics and responded well. Follow-up time at present is 5.5 years for the first, and 4.5 years for the second, patient.

  9. In-situ infiltration performance of different permeable pavements in a employee used parking lot--A four-year study.

    Science.gov (United States)

    Kumar, Kuldip; Kozak, Joseph; Hundal, Lakhwinder; Cox, Albert; Zhang, Heng; Granato, Thomas

    2016-02-01

    Permeable pavements are being adopted as a green solution in many parts of the world to manage urban stormwater quantity and quality. This paper reports on the measured in-situ infiltration performance over a four-year period since construction and use of three permeable parking sections (permeable pavers, permeable concrete and permeable asphalt) of an employee car parking lot. There was only a marginal decline in infiltration rates of all three pavements after one year of use. However, between years two to four, the infiltration rates declined significantly due to clogging of pores either by dry deposition of particles and/or shear stress of vehicles driving and degrading the permeable surfaces; during the last two years, a greater decline was also observed in driving areas of the parking lots compared to parking slots, where minimal wear and tear are expected. Maintenance strategies were employed to reclaim some of the lost infiltration rate of the permeable pavements to limited success. Despite this decline, the infiltration rates were still four to five times higher than average rainstorm intensity in the region. Thus, these permeable pavement parking lots may have significant ecological importance due to their ability to infiltrate rainwater quickly, reduce the runoff in the catchment area, and also dampen runoff peak flows that could otherwise enter the collection system for treatment in a combined sewer area.

  10. Timeline trend profile and seasonal variations in nicotine present in ambient PM10 samples: A four year investigation from Delhi region, India

    Science.gov (United States)

    Yadav, Shweta; Tandon, Ankit; Attri, Arun K.

    2014-12-01

    The detection of nicotine, an organic tracer for Environmental Tobacco Smoke (ETS), in the collected PM10 samples from Delhi region's ambient environment, in a appropriately designed investigation was initiated over four years (2006-2009) to: (1) Comprehend seasonal and inter-annual variations in the nicotine present in PM10; (2) Extract regression based linear trend profile manifested by nicotine in PM10; (3) Determine the non-linear trend timeline from the nicotine data, and compare it with the obtained linear trend; (4) Suggest the possible use of the designed experiment and analysis to have a qualitative appraisal of Tobacco Smoking activity in the sampling region. The PM10 samples were collected in a monthly time-series sequence at a known receptor site. Quantitative estimates of nicotine (ng m-3) were made by using a Thermal Desorption Gas Chromatography Mass Spectrometry (TD-GC/MS). The annual average concentrations of nicotine (ng m-3) were 516 ± 302 (2008) > 494 ± 301 (2009) > 438 ± 250 (2007) > 325 ± 149 (2006). The estimated linear trend of 5.4 ng m-3 month-1 corresponded to 16.3% per annum increase in the PM10 associated nicotine. The industrial production of India's tobacco index normalized to Delhi region's consumption, pegged an increase at 10.5% per annum over this period.

  11. AIR QUALITY AND VARIATIONS IN PM10 POLLUTANT CONCENTRATION IN WESTERN IRAN DURING A FOUR -YEAR PERIOD (2008-2011, KERMANSHAH - A CASE STUDY

    Directory of Open Access Journals (Sweden)

    KIOMARS SHARAFI

    2015-01-01

    Full Text Available The present study explored the quality of PM10, over a four-year period in the western part of Iran. A total of 1334 samples were collected from air pollution measurement stations in Kermanshah. Statistical analysis was carried out using the SPSS ver.14. The results showed that the total percent change in PM10 over a period of 4 years was 10.04%, 59.2%, 26.4%, 0.7% and 3.3%. The highest and the lowest frequencies of PM10 occurred in February with 37.7%, June and July with 0%; winter and summer months with 21.96 and 4.81%, respectively. The results are depicted that the air quality in July, the summer months and 2008 was worst in terms of PM10 concentration, which was mainly due to the fluctuations and acute entrance of dust particles into western Iran. Hence, the reduction in such emissions is indispensable and requires extensive and considerable cooperation between the government of Iran and the neighbouring countries.

  12. Impact on practice of a British Association for Sexual Health and HIV Sexually Transmitted Infections Foundation (STIF) course: an audit of the first four years in Ireland.

    Science.gov (United States)

    Calamai, A; Howard, R; Kelly, R; Lambert, J

    2013-02-01

    In order to investigate the overall impact of the British Association for Sexual Health and HIV (BASHH) Sexually Transmitted Infections Foundation (STIF) course taught in Ireland since 2007, attendees were sent two questionnaires to investigate the overall impact of the course, its effect on clinical practice and the need for further education. Response rate was 19.4%. The majority found the course beneficial and that it did cover their practice needs (96.4%), with 83.6% saying that their confidence and technique in sexual history taking had improved. There was a 3.7% increase in the provision of HIV testing from precourse levels, although only 80% did so routinely; a 12.7% increase in syphilis testing; a 5.4% increase in testing for Chlamydia and a 12.7% increase for gonorrhoea. Some confusion seems to persist in relation to sexually transmitted infection (STI) risk factors. The second questionnaire tested STI knowledge. Most respondents scored well (average 81% correct answers); however, respondents who attended four years previously scored, on average, 7% worse than the others, suggesting the need for a periodic update in the area of STI education.

  13. A characterization of intermediate-scale spread F structure from four years of high-resolution C/NOFS satellite data

    Science.gov (United States)

    Rino, Charles L.; Carrano, Charles S.; Groves, Keith M.; Roddy, Patrick A.

    2016-06-01

    Power law spectra have been invoked to interpret equatorial scintillation data for decades. Published analyses of intensity and phase scintillation data typically report power law spectra of the form q-p with 2.4 power law components. Strong scatter simulations and recent theoretical results have shown that two-component power law spectra can reconcile simultaneous equatorial scintillation observations from VHF to S-Band. The Communication/Navigation Outage Forecasting System (C/NOFS) satellite Planar Langmuir Probe generated a multiyear high-resolution sampling of equatorial spread F, but published analyses to date have reported only single-component power laws over scales from tens of kilometers to 70 m. This paper summarizes the analysis of high-resolution C/NOFS data collected over the four year period 2011 to 2014. Following an earlier investigation of several months of C/NOFS data by the authors of this paper, the extended data set revealed a pattern of occurrence of two-component spectra in the most highly disturbed data sets. The results confirm a known inverse correlation between turbulent strength and spectral index. The new results are interpreted as an equatorial spread F life cycle pattern with two-component spectra in the early development phase giving way to single-component spectra in the decay phase.

  14. The prevalence of Giardia intestinalis and Entamoeba histolytica/dispar in Van Regional Training and Research Hospital: A four-year monitoring

    Directory of Open Access Journals (Sweden)

    Yasemin Bayram

    2013-03-01

    Full Text Available Objective: The aim of this retrospective study was to determinethe frequency rates of Giardia and E.histolytica/E.dispar and their distribution by years as well as agegroups and gender distribution during a four-year period.Materials and methods: A total of 9911 stool samplessent to our laboratory between January 2008 and December2011 were tested for parasites. Native-Lugol and formolethyl acetate sedimentation methods were utilized formicroscopic identification of Entamoeba sp. and Giardiatrophozoites and cysts in fresh stool samples. Additionally,trichrome staining was performed in stool sampleswhere the distinctive diagnosis could not be confirmed.Results: From a total of 9911 stool samples analyzedduring the study, 4.7% were positive for Giardia and 6.2%were positive for Entemoeba histolytica/ Entemoeba dispar.Of Giardia-positive patients 57% were male and 43%female. Similarly, 56% of Entemoeba histolytica/ Entemoebadispar positive patients were male and 44% werefemale. Both parasites’ higher frequency rates seen inmale groups were found statistically significant (p<0.01.Conclusion: It is seen that intestinal protozoon infectionsare still present as an important public health problemin our region. In order to prevent this problem, personalhygiene and sanitation rules education for community aswell as infrastructure improvements are necessary.Key words: Giardia, Entemoeba histolytica/ Entemoeba dispar, prevalence

  15. Evaluation of the use of electronic health data to classify four-year mortality risk for older adults undergoing screening colonoscopies.

    Science.gov (United States)

    Synnestvedt, Marie B; Weiner, Mark G

    2011-01-01

    Current cancer screening recommendations often apply coarse age cutoffs for screening requirements without regard to predicted life expectancy. Using these cutoffs, healthier older patients may be under-screened, and sicker younger patients may be screened too often. Mortality risk classification using EHR data could be used to tailor screening reminders to physicians in ways that better align screening recommendations with patients who are more likely to live long enough to benefit from early detection. We have evaluated the performance of an existing prognostic index for 4-year mortality using data readily available in the electronic health record (EHR), and investigated the effect of the index in retrospective cohorts of adults age 65 and older undergoing screening colonoscopy. Risk scores in this adaptation of a four-year prognostic index were found to be associated with actual death rates and consistent with mortality rates from a national sample. Our results demonstrate that data extracted from electronic health records can be used to classify mortality risk. With improvements, including extension to a 5-year mortality model with inclusion of additional variables and extension of variable definitions, informatics methods to implement mortality models may prove to be clinically useful in tailoring screening guidelines.

  16. Findings of a Four-Year Randomized Controlled Clinical Trial Comparing Two-Piece and One-Piece Zirconia Abutments Supporting Single Prosthetic Restorations in Maxillary Anterior Region

    Directory of Open Access Journals (Sweden)

    Guerino Paolantoni

    2016-01-01

    Full Text Available The purpose of this randomized controlled study is to investigate the clinical results obtained over four years and incidence of complications associated with one- versus two-piece custom made zirconia anchorages, in single tooth implant-supported restorations of the maxillary anterior region. Sixty-five patients, with a total of 74 missing maxillary teeth, were selected in the period from February 2007 to July 2010. Two different ways of custom made zirconia abutment and final prosthetic restoration were evaluated: a standard zirconia abutment associated with a pressed layer of lithium disilicate with an all-ceramic cemented restoration versus one-piece restoration with the facing porcelain fired and pressed straight to the custom made zirconia abutment. In 29 cases, the restoration consisted of an all-ceramic restoration for cementation (two pieces; in 45 cases the restoration was a screw-retained restoration (one piece. Three all-ceramic restorations broke during the observation time. Two one-piece restorations fractured after 26 months. At follow-up examination there were no significant differences between one-piece and two-piece groups regarding the PI, BI, and MBL. Awaiting studies with longer follow-up times, a careful conclusion is that zirconia anchorages for single-implant restorations seem to demonstrate good short-term technical and biological results.

  17. A long-term predictive validity study: can the CDI Short Form be used to predict language and early literacy skills four years later?

    Science.gov (United States)

    Can, Dilara Deniz; Ginsburg-Block, Marika; Golinkoff, Roberta Michnick; Hirsh-Pasek, Kathryn

    2013-09-01

    This longitudinal study examined the predictive validity of the MacArthur Communicative Developmental Inventories-Short Form (CDI-SF), a parent report questionnaire about children's language development (Fenson, Pethick, Renda, Cox, Dale & Reznick, 2000). Data were first gathered from parents on the CDI-SF vocabulary scores for seventy-six children (mean age=1 ; 10). Four years later (mean age=6 ; 1), children were assessed on language outcomes (expressive vocabulary, syntax, semantics and pragmatics) and code-related skills, including phonemic awareness, word recognition and decoding skills. Hierarchical regression analyses revealed that early expressive vocabulary accounted for 17% of the variance in picture vocabulary, 11% of the variance in syntax, and 7% of the variance in semantics, while not accounting for any variance in pragmatics in kindergarten. CDI-SF scores did not predict code-related skills in kindergarten. The importance of early vocabulary skills for later language development and CDI-SF as a valuable research tool are discussed.

  18. A hacker's guide to catching a debris flow: Lessons learned from four years of chasing mud in Colorado and southern California

    Science.gov (United States)

    Kean, J. W.; McCoy, S. W.; Staley, D. M.; Coe, J.; Leeper, R.; Tucker, G. E.

    2012-12-01

    Direct measurements of natural debris flows provide valuable insights into debris-flow processes and hazards. Yet debris flows are difficult to "catch" because they live in rugged terrain, appear infrequently, and have an appetite for destroying monitoring equipment. We present an overview of some successful (and failed) techniques we have used over the past four years to obtain direct measurements of 40+ debris flows in Colorado and southern California. Following the "MacGyver" theme of the session, we focus on the improvised equipment and methods we use in our hunt for quality data. These include an inexpensive erosion sensor to measure rates of debris-flow entrainment, a custom load cell enclosure for measuring debris-flow normal force, tracer rocks implanted with passive integrated transponders, basic pressure transducers to measure debris-flow timing, and standard digital cameras adapted to obtain high-resolution (1936 x 1288 pixels) video footage of debris flows. These techniques are also suitable for catching data on elusive flash floods. In addition, we also share some practical solutions to the logistical problems associated with installing monitoring equipment in rugged debris-flow terrain, such as suspension of non-contact stage gages high above channels.

  19. Normal proliferation and differentiation of Hoxc-8 transgenic chondrocytes in vitro

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    Mello Maria

    2003-04-01

    Full Text Available Abstract Background Hox genes encode transcription factors that are involved in pattern formation in the skeleton, and recent evidence suggests that they also play a role in the regulation of endochondral ossification. To analyze the role of Hoxc-8 in this process in more detail, we applied in vitro culture systems, using high density cultures of primary chondrocytes from neonatal mouse ribs. Results Cultured cells were characterized on the basis of morphology (light microscopy and production of cartilage-specific extracellular matrix (sulfated proteoglycans and type II Collagen. Hypertrophy was demonstrated by increase in cell size, alkaline phosphatase activity and type X Collagen immunohistochemistry. Proliferation was assessed by BrdU uptake and flow cytometry. Unexpectedly, chondrocytes from Hoxc-8 transgenic mice, which exhibit delayed cartilage maturation in vivo 1, were able to proliferate and differentiate normally in our culture systems. This was the case even though freshly isolated Hoxc-8 transgenic chondrocytes exhibited significant molecular differences as measured by real-time quantitative PCR. Conclusions The results demonstrate that primary rib chondrocytes behave similar to published reports for chondrocytes from other sources, validating in vitro approaches for studies of Hox genes in the regulation of endochondral ossification. Our analysis of cartilage-producing cells from Hoxc-8 transgenic mice provides evidence that the cellular phenotype induced by Hoxc-8 overexpression in vivo is reversible in vitro.

  20. Mefloquine inhibits chondrocytic proliferation by arresting cell cycle in G2/M phase.

    Science.gov (United States)

    Li, Qiong; Chen, Zeng-Gan; Xia, Qing; Lin, Jian-Ping; Yan, Zuo-Qin; Yao, Zheng-Jun; Dong, Jian

    2015-01-01

    Mefloquine (MQ), an analog of chloroquine, exhibits a promising cytotoxic activity against carcinoma cell lines and for the treatment of glioblastoma patients. The present study demonstrates the effect of mefloquine on proliferation and cell cycle in chondrocytes. MTT assay and propidium iodide staining were used for the analysis of proliferation and cell cycle distribution, respectively. Western blot analysis was used to examine the expression levels of cyclin B1/cdc2, cdc25c, p21WAF1/CIP1 and p53. The results revealed that mefloquine inhibited the proliferation of chondrocytes and caused cell cycle arrests in the G2/M phase. The proliferation of chondrocytes was reduced to 27% at 40 μM concentration of mefloquine after 48 h. The population of chondrocytes in G2/M phase was found to be 15.7 and 48.4%, respectively at 10 and 40 μM concentration of mefloquine at 48 h following treatment. The expression of the cell cycle regulatory proteins including, cyclin B1/cdc2 and cdc25c was inhibited. On the other hand, mefloquine treatment promoted the expression of p21WAF1/CIP1 and p53 at 40 μM concentration after 48 h. Therefore, mefloquine inhibits proliferation and induces cell cycle arrest in chondrocytes.

  1. Single Cell Confocal Raman Spectroscopy of Human Osteoarthritic Chondrocytes: A Preliminary Study

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    Rajesh Kumar

    2015-04-01

    Full Text Available A great deal of effort has been focused on exploring the underlying molecular mechanism of osteoarthritis (OA especially at the cellular level. We report a confocal Raman spectroscopic investigation on human osteoarthritic chondrocytes. The objective of this investigation is to identify molecular features and the stage of OA based on the spectral signatures corresponding to bio-molecular changes at the cellular level in chondrocytes. In this study, we isolated chondrocytes from human osteoarthritic cartilage and acquired Raman spectra from single cells. Major spectral differences between the cells obtained from different International Cartilage Repair Society (ICRS grades of osteoarthritic cartilage were identified. During progression of OA, a decrease in protein content and an increase in cell death were observed from the vibrational spectra. Principal component analysis and subsequent cross-validation was able to associate osteoarthritic chondrocytes to ICRS Grade I, II and III with specificity 100.0%, 98.1%, and 90.7% respectively, while, sensitivity was 98.6%, 82.8%, and 97.5% respectively. The overall predictive efficiency was 92.2%. Our pilot study encourages further use of Raman spectroscopy as a noninvasive and label free technique for revealing molecular features associated with osteoarthritic chondrocytes.

  2. Mechanical vibrations increase the proliferation of articular chondrocytes in high-density culture.

    Science.gov (United States)

    Kaupp, J A; Waldman, S D

    2008-07-01

    Tissue engineering is a promising approach for articular cartilage repair; however, it still has proven a challenge to produce tissue from the limited number of cells that can be extracted from a single individual. Relatively few cell expansion methods exist without the problems of dedifferentiation and/or loss of potency. Previously, it has been shown that mechanical vibrations can enhance chondrocyte proliferation in monolayer culture. Thus, it was hypothesized that chondrocytes grown in high-density culture would respond in a similar fashion while maintaining phenotypic stability. Isolated bovine articular chondrocytes were seeded in high-density culture on Millicell filters and subjected to mechanical vibrations 48 h after seeding. Mechanical vibrations enhanced chondrocyte proliferation at frequencies above 350 Hz, with the peak response occurring at a 1g amplitude for a duration of 30 min. Under these conditions, the gene expression of cartilage-specific and dedifferentiation markers (collagen II, collagen I, and aggrecan) were unchanged by the imposed stimulus. To determine the effect of accumulated extracellular matrix (ECM) on this proliferative response, selected cultures were stimulated under the same conditions after varying lengths of preculture. The amount of accumulated ECM (collagen and proteoglycans) decreased this proliferative response, with the cultures becoming insensitive to the stimulus after 1 week of preculture. Thus, mechanical vibration can serve as an effective means preferentially to stimulate the proliferation of chondrocytes during culture, but its effects appear to be limited to the early stages where ECM accumulation is at a minimum.

  3. Combining Targeted Metabolomic Data with a Model of Glucose Metabolism: Toward Progress in Chondrocyte Mechanotransduction

    Science.gov (United States)

    Salinas, Daniel; Carlson, Ross P.; McCutchen, Carley N.

    2017-01-01

    Osteoarthritis is a debilitating disease likely involving altered metabolism of the chondrocytes in articular cartilage. Chondrocytes can respond metabolically to mechanical loads via cellular mechanotransduction, and metabolic changes are significant because they produce the precursors to the tissue matrix necessary for cartilage health. However, a comprehensive understanding of how energy metabolism changes with loading remains elusive. To improve our understanding of chondrocyte mechanotransduction, we developed a computational model to calculate the rate of reactions (i.e. flux) across multiple components of central energy metabolism based on experimental data. We calculated average reaction flux profiles of central metabolism for SW1353 human chondrocytes subjected to dynamic compression for 30 minutes. The profiles were obtained solving a bounded variable linear least squares problem, representing the stoichiometry of human central energy metabolism. Compression synchronized chondrocyte energy metabolism. These data are consistent with dynamic compression inducing early time changes in central energy metabolism geared towards more active protein synthesis. Furthermore, this analysis demonstrates the utility of combining targeted metabolomic data with a computational model to enable rapid analysis of cellular energy utilization. PMID:28056047

  4. Mechanotransduction in primary human osteoarthritic chondrocytes is mediated by metabolism of energy, lipids, and amino acids.

    Science.gov (United States)

    Zignego, Donald L; Hilmer, Jonathan K; June, Ronald K

    2015-12-16

    Chondrocytes are the sole cell type found in articular cartilage and are repeatedly subjected to mechanical loading in vivo. We hypothesized that physiological dynamic compression results in changes in energy metabolism to produce proteins for maintenance of the pericellular and extracellular matrices. The objective of this study was to develop an in-depth understanding for the short term (human chondrocytes harvested from femoral heads of osteoarthritic donors. Cell-seeded agarose constructs were randomly assigned to experimental groups, and dynamic compression was applied for 0, 15, or 30min. Following dynamic compression, metabolites were extracted and detected by HPLC-MS. Untargeted analyzes examined changes in global metabolomics profiles and targeted analysis examined the expression of specific metabolites related to central energy metabolism. We identified hundreds of metabolites that were regulated by applied compression, and we report the detection of 16 molecules not found in existing metabolite databases. We observed patient-specific mechanotransduction with aging dependence. Targeted studies found a transient increase in the ratio of NADP+ to NADPH and an initial decrease in the ratio of GDP to GTP, suggesting a flux of energy into the TCA cycle. By characterizing metabolomics profiles of primary chondrocytes in response to applied dynamic compression, this study provides insight into how OA chondrocytes respond to mechanical load. These results are consistent with increases in glycolytic energy utilization by mechanically induced signaling, and add substantial new data to a complex picture of how chondrocytes transduce mechanical loads.

  5. Hyaluronic acid abrogates nitric oxide-dependent stimulation of collagen degradation in cultured human chondrocytes.

    Science.gov (United States)

    Surazynski, Arkadiusz; Miltyk, Wojciech; Czarnomysy, Robert; Grabowska, Joanna; Palka, Jerzy

    2009-07-01

    Experimental inflammation induced in cultured chondrocytes by inflammatory cytokine IL-1 beta stimulates collagen degradation by metalloproteinases. We propose that nitric oxide (NO) may represent down stream signaling molecule of IL-1-induced collagen degradation in chondrocytes. It was found that IL-1 beta induced the activity of MMP-2 and MMP-9 during the 48 h time course of the experiment, especially after 24h incubation, while DETA/NO, donor of NO, stimulated the process at 12h incubation. The mechanism of IL-1-dependent stimulation of NO production was found at the level of iNOS expression and activation of NF-kappaB. We found that hyaluronic acid (HA) counteracted IL-induced degradation of collagen in chondrocytes. Although, HA by itself had no effect on the metaloproteinases activity, when added to IL-1 beta or DETA/NO treated chondrocytes it contributed to the restoration of the MMPs activity to the control level. The mechanism of this phenomenon involves inhibition of NF-kappaB activation. The data suggest that NO may represent a target molecule for protective effect of hyaluronic acid on interleukin-1-induced stimulation of metaloproteinases activity in cultured human chondrocytes.

  6. Chondrocytes, Mesenchymal Stem Cells, and Their Combination in Articular Cartilage Regenerative Medicine.

    Science.gov (United States)

    Nazempour, A; Van Wie, B J

    2016-05-01

    Articular cartilage (AC) is a highly organized connective tissue lining, covering the ends of bones within articulating joints. Its highly ordered structure is essential for stable motion and provides a frictionless surface easing load transfer. AC is vulnerable to lesions and, because it is aneural and avascular, it has limited self-repair potential which often leads to osteoarthritis. To date, no fully successful treatment for osteoarthritis has been reported. Thus, the development of innovative therapeutic approaches is desperately needed. Autologous chondrocyte implantation, the only cell-based surgical intervention approved in the United States for treating cartilage defects, has limitations because of de-differentiation of articular chondrocytes (AChs) upon in vitro expansion. De-differentiation can be abated if initial populations of AChs are co-cultured with mesenchymal stem cells (MSCs), which not only undergo chondrogenesis themselves but also support chondrocyte vitality. In this review we summarize studies utilizing AChs, non-AChs, and MSCs and compare associated outcomes. Moreover, a comprehensive set of recent human studies using chondrocytes to direct MSC differentiation, MSCs to support chondrocyte re-differentiation and proliferation in co-culture environments, and exploratory animal intra- and inter-species studies are systematically reviewed and discussed in an innovative manner allowing side-by-side comparisons of protocols and outcomes. Finally, a comprehensive set of recommendations are made for future studies.

  7. The Regulatory Role of Signaling Crosstalk in Hypertrophy of MSCs and Human Articular Chondrocytes.

    Science.gov (United States)

    Zhong, Leilei; Huang, Xiaobin; Karperien, Marcel; Post, Janine N

    2015-08-14

    Hypertrophic differentiation of chondrocytes is a main barrier in application of mesenchymal stem cells (MSCs) for cartilage repair. In addition, hypertrophy occurs occasionally in osteoarthritis (OA). Here we provide a comprehensive review on recent literature describing signal pathways in the hypertrophy of MSCs-derived in vitro differentiated chondrocytes and chondrocytes, with an emphasis on the crosstalk between these pathways. Insight into the exact regulation of hypertrophy by the signaling network is necessary for the efficient application of MSCs for articular cartilage repair and for developing novel strategies for curing OA. We focus on articles describing the role of the main signaling pathways in regulating chondrocyte hypertrophy-like changes. Most studies report hypertrophic differentiation in chondrogenesis of MSCs, in both human OA and experimental OA. Chondrocyte hypertrophy is not under the strict control of a single pathway but appears to be regulated by an intricately regulated network of multiple signaling pathways, such as WNT, Bone morphogenetic protein (BMP)/Transforming growth factor-β (TGFβ), Parathyroid hormone-related peptide (PTHrP), Indian hedgehog (IHH), Fibroblast growth factor (FGF), Insulin like growth factor (IGF) and Hypoxia-inducible factor (HIF). This comprehensive review describes how this intricate signaling network influences tissue-engineering applications of MSCs in articular cartilage (AC) repair, and improves understanding of the disease stages and cellular responses within an OA articular joint.

  8. Lubricin is expressed in chondrocytes derived from osteoarthritic cartilage encapsulated in poly (ethylene glycol) diacrylate scaffold.

    Science.gov (United States)

    Musumeci, G; Loreto, C; Carnazza, M L; Coppolino, F; Cardile, V; Leonardi, R

    2011-01-01

    Osteoarthritis (OA) is characterized by degenerative changes within joints that involved quantitative and/or qualitative alterations of cartilage and synovial fluid lubricin, a mucinous glycoprotein secreted by synovial fibroblasts and chondrocytes. Modern therapeutic methods, including tissue-engineering techniques, have been used to treat mechanical damage of the articular cartilage but to date there is no specific and effective treatment. This study aimed at investigating lubricin immunohistochemical expression in cartilage explant from normal and OA patients and in cartilage constructions formed by Poly (ethylene glycol) (PEG) based hydrogels (PEG-DA) encapsulated OA chondrocytes. The expression levels of lubricin were studied by immunohistochemistry: i) in tissue explanted from OA and normal human cartilage; ii) in chondrocytes encapsulated in hydrogel PEGDA from OA and normal human cartilage. Moreover, immunocytochemical and western blot analysis were performed in monolayer cells from OA and normal cartilage. The results showed an increased expression of lubricin in explanted tissue and in monolayer cells from normal cartilage, and a decreased expression of lubricin in OA cartilage. The chondrocytes from OA cartilage after 5 weeks of culture in hydrogels (PEGDA) showed an increased expression of lubricin compared with the control cartilage. The present study demonstrated that OA chondrocytes encapsulated in PEGDA, grown in the scaffold and were able to restore lubricin biosynthesis. Thus our results suggest the possibility of applying autologous cell transplantation in conjunction with scaffold materials for repairing cartilage lesions in patients with OA to reduce at least the progression of the disease.

  9. Adiponectin and leptin induce VCAM-1 expression in human and murine chondrocytes.

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    Javier Conde

    Full Text Available BACKGROUND: Osteoarthritis (OA and rheumatoid arthritis (RA, the most common rheumatic diseases, are characterized by irreversible degeneration of the joint tissues. There are several factors involved in the pathogenesis of these diseases including pro-inflammatory cytokines, adipokines and adhesion molecules. OBJECTIVE: Up to now, the relationship between adipokines and adhesion molecules at cartilage level was not explored. Thus, the aim of this article was to study the effect of leptin and adiponectin on the expression of VCAM-1 in human and murine chondrocytes. For completeness, intracellular signal transduction pathway was also explored. METHODS: VCAM-1 expression was assessed by quantitative RT-PCR and western blot analysis upon treatment with leptin, adiponectin and other pertinent reagents in cultured human primary chondrocytes. Signal transduction pathways have been explored by using specific pharmacological inhibitors in the adipokine-stimulated human primary chondrocytes and ATDC5 murine chondrocyte cell line. RESULTS: Herein, we demonstrate, for the first time, that leptin and adiponectin increase VCAM-1 expression in human and murine chondrocytes. In addition, both adipokines have additive effect with IL-1β. Finally, we demonstrate that several kinases, including JAK2, PI3K and AMPK are at a play in the intracellular signalling of VCAM-1 induction. CONCLUSIONS: Taken together, our results suggest that leptin and adiponectin could perpetuate cartilage-degrading processes by inducing also factors responsible of leukocyte and monocyte infiltration at inflamed joints.

  10. Expression of Transient Receptor Potential Vanilloid (TRPV Channels in Different Passages of Articular Chondrocytes

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    Richard Barrett-Jolley

    2012-04-01

    Full Text Available Ion channels play important roles in chondrocyte mechanotransduction. The transient receptor potential vanilloid (TRPV subfamily of ion channels consists of six members. TRPV1-4 are temperature sensitive calcium-permeable, relatively non-selective cation channels whereas TRPV5 and TRPV6 show high selectivity for calcium over other cations. In this study we investigated the effect of time in culture and passage number on the expression of TRPV4, TRPV5 and TRPV6 in articular chondrocytes isolated from equine metacarpophalangeal joints. Polyclonal antibodies raised against TRPV4, TRPV5 and TRPV6 were used to compare the expression of these channels in lysates from first expansion chondrocytes (P0 and cells from passages 1–3 (P1, P2 and P3 by western blotting. TRPV4, TRPV5 and TRPV6 were expressed in all passages examined. Immunohistochemistry and immunofluorescence confirmed the presence of these channels in sections of formalin fixed articular cartilage and monolayer cultures of methanol fixed P2 chondrocytes. TRPV5 and TRPV6 were upregulated with time and passage in culture suggesting that a shift in the phenotype of the cells in monolayer culture alters the expression of these channels. In conclusion, several TRPV channels are likely to be involved in calcium signaling and homeostasis in chondrocytes.

  11. Suction blister grafting - Modifications for easy harvesting and grafting

    Directory of Open Access Journals (Sweden)

    2012-01-01

    Full Text Available Suction blister grafting is a simple modality of treatment of patients with resistant and stable vitiligo. But raising the blisters may be time consuming and transferring to the recipient site may be difficult as the graft is ultrathin. By doing some modifications we can make the technique simpler and easier. We can decrease the blister induction time by intradermal injection of saline, exposure to Wood′s lamp, intrablister injection of saline. By these methods we can decrease the blister induction time from 2-3 hrs to 45-90 minutes. After harvesting the graft, it can be transferred to the recipient area by taking the graft on a sterile glass slide, on the gloved finger, rolling the graft over a sterile syringe and then spreading on the recipient area, or taking on the sterile wrapper of paraffin dressing and then placing over the recipient area.

  12. Fibroblast growth factor-1 is a mesenchymal stromal cell-secreted factor stimulating proliferation of osteoarthritic chondrocytes in co-culture

    NARCIS (Netherlands)

    Wu, Ling; Leijten, Jeroen; Blitterswijk, van Clemens A.; Karperien, Marcel

    2013-01-01

    Previously, we showed that mesenchymal stromal cells (MSCs) in co-culture with primary chondrocytes secrete soluble factors that increase chondrocyte proliferation. The objective of this study is to identify these factors. Human primary chondrocytes (hPCs) isolated from late-stage osteoarthritis pat

  13. Adhesion-mediated signal transduction in human articular chondrocytes: the influence of biomaterial chemistry and tenascin-C

    NARCIS (Netherlands)

    Mahmood, Tahir A.; Jong, de Ruben; Riesle, Jens; Langer, Robert; Blitterswijk, van Clemens A.

    2004-01-01

    Chondrocyte ‘dedifferentiation’ involves the switching of the cell phenotype to one that no longer secretes extracellular matrix found in normal cartilage and occurs frequently during chondrocyte expansion in culture. It is also characterized by the differential expression of receptors and intracell

  14. Differences in Cartilage-Forming Capacity of Expanded Human Chondrocytes From Ear and Nose and Their Gene Expression Profiles

    NARCIS (Netherlands)

    Hellingman, C.A.; Verwiel, E.T.P.; Slagt, I.; Koevoet, W.; Poublon, R.M.L.; Nolst-Trenite, G.J.; de Jong, R.J.B.; Jahr, H.; van Osch, G.J.V.M.

    2011-01-01

    The aim of this study was to evaluate the potential of culture-expanded human auricular and nasoseptal chondrocytes as cell source for regeneration of stable cartilage and to analyze the differences in gene expression profile of expanded chondrocytes from these specific locations. Auricular chondroc

  15. Graft selection in cerebral revascularization.

    Science.gov (United States)

    Baaj, Ali A; Agazzi, Siviero; van Loveren, Harry

    2009-05-01

    Cerebral revascularization constitutes an important treatment modality in the management of complex aneurysms, carotid occlusion, tumor, and moyamoya disease. Graft selection is a critical step in the planning of revascularization surgery, and depends on an understanding of graft and regional hemodynamics, accessibility, and patency rates. The goal of this review is to highlight some of these properties.

  16. Polyether/Polyester Graft Copolymers

    Science.gov (United States)

    Bell, Vernon L., Jr.; Wakelyn, N.; Stoakley, D. M.; Proctor, K. M.

    1986-01-01

    Higher solvent resistance achieved along with lower melting temperature. New technique provides method of preparing copolymers with polypivalolactone segments grafted onto poly (2,6-dimethyl-phenylene oxide) backbone. Process makes strong materials with improved solvent resistance and crystalline, thermally-reversible crosslinks. Resulting graft copolymers easier to fabricate into useful articles, including thin films, sheets, fibers, foams, laminates, and moldings.

  17. Stress relaxation analysis of single chondrocytes using porohyperelastic model based on AFM experiments

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    Trung Dung Nguyen

    2014-01-01

    Full Text Available Based on atomic force microscopytechnique, we found that the chondrocytes exhibits stress relaxation behavior. We explored the mechanism of this stress relaxation behavior and concluded that the intracellular fluid exuding out from the cells during deformation plays the most important role in the stress relaxation. We applied the inverse finite element analysis technique to determine necessary material parameters for porohyperelastic (PHE model to simulate stress relaxation behavior as this model is proven capable of capturing the non-linear behavior and the fluid-solid interaction during the stress relaxation of the single chondrocytes. It is observed that PHE model can precisely capture the stress relaxation behavior of single chondrocytes and would be a suitable model for cell biomechanics.

  18. Andrographolide Enhances Proliferation and Prevents Dedifferentiation of Rabbit Articular Chondrocytes: An In Vitro Study

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    Li-ke Luo

    2015-01-01

    Full Text Available As the main active constituent of Andrographis paniculata that was applied in treatment of many diseases including inflammation in ancient China, andrographolide (ANDRO was found to facilitate reduction of edema and analgesia in arthritis. This suggested that ANDRO may be promising anti-inflammatory agent to relieve destruction and degeneration of cartilage after inflammation. In this study, the effect of ANDRO on rabbit articular chondrocytes in vitro was investigated. Results showed that not more than 8 μM ANDRO did no harm to chondrocytes (P0.05. The viability assay, hematoxylin-eosin, safranin O, and immunohistochemical staining also showed better performances in ANDRO groups. As to the doses, 3 μM ANDRO showed the best performance. The results indicate that ANDRO can accelerate proliferation of rabbit articular chondrocytes in vitro and meanwhile maintain the phenotype, which may provide valuable references for further exploration on arthritis.

  19. Diabetic nephropathy as a cause of end-stage renal disease in Tabuk area, Saudi Arabia: A four-year study

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    Osama El Minshawy

    2014-01-01

    Full Text Available Diabetic nephropathy (DN as a cause of end-stage renal disease (ESRD is increa-sing worldwide. In some countries, it is the most common cause of ESRD. Our objective was to assess the incidence of DN as a cause of ESRD in Tabuk, to evaluate its changes in four years, and to compare the data of Tabuk with data from the United States (US to be aware of factors causing the difference. Data of ESRD patients with DN treated with renal replacement therapy (RRT was evaluated from 2009 to 2012. RRT was defined as ESRD patients who were treated either with chronic regular hemodialysis (HD, renal transplantation (Tx or continuous ambulatory peritoneal dialysis (PD. The incidence of DN as a cause of ESRD increased from 8% in 2009 to 18% in 2012. The mean age of this group was significantly higher than in patients on RRT due to other etiologies. Also, DN was more widespread in built-up areas than pastoral areas. The mortality rate decreased from 20% in 2009 to 14% in 2012. Despite this decrease, the mortality rate was still higher than that in patients on RRT due to other etiologies. When we restricted our analysis to patients treated by HD (76%, Tx (17% or PD (7%, the results were not significantly different. DN in the Tabuk area is rising, but is less widespread than in the US possibly because of an increased occurrence of other causes of ESRD or early loss of diabetic patients. Therefore, careful management of diabetic patients is obligatory.

  20. Exploration and Practice of Four -year Vocational Education%四年制职业教育的探索与实践

    Institute of Scientific and Technical Information of China (English)

    孙小红

    2015-01-01

    国家社会经济发展和构建现代职业教育体系对高端技能型人才的培养提出了迫切的要求。本文在广泛调研、分析的基础上,确定了四年制高职的人才培养目标和课程体系,以本科与高职院校联合培养应用型、高端技能型人才试点班为契机,对组织保证、学业导师、教学管理和监督等创新性教学管理模式进行了有益的尝试,将人才培养方案转化成培养成果,为高层次职业教育开创了先河。%the social and economic development of the state and the construction of modern voca‐tional education system impose urgent requirements for cultivating the high-end skilled personnel .This paper defines the objectives and curricular of cultivating four -year higher vocational personnel through comprehensive investigation and research .It takes the opportunities of jointly cultivating practical and high-end skilled personnel by universities and higher vocational colleges to make rewarding attempts in organizational guarantee , academic advisor , teaching management ,supervision and other innovative teaching management patterns and converts the personnel training program to the training results and creates the precedent for higher vocational education .

  1. Pediatric Epidemic of Salmonella enterica Serovar Typhimurium in the Area of L’Aquila, Italy, Four Years after a Catastrophic Earthquake

    Directory of Open Access Journals (Sweden)

    Giovanni Nigro

    2016-05-01

    Full Text Available Background: A Salmonella enterica epidemic occurred in children of the area of L’Aquila (Central Italy, Abruzzo region between June 2013 and October 2014, four years after the catastrophic earthquake of 6 April 2009. Methods: Clinical and laboratory data were collected from hospitalized and ambulatory children. Routine investigations for Salmonella infection were carried out on numerous alimentary matrices of animal origin and sampling sources for drinking water of the L’Aquila district, including pickup points of the two main aqueducts. Results: Salmonella infection occurred in 155 children (83 females: 53%, aged 1 to 15 years (mean 2.10. Of these, 44 children (28.4% were hospitalized because of severe dehydration, electrolyte abnormalities, and fever resistant to oral antipyretic and antibiotic drugs. Three children (1.9% were reinfected within four months after primary infection by the same Salmonella strain. Four children (2.6%, aged one to two years, were coinfected by rotavirus. A seven-year old child had a concomitant right hip joint arthritis. The isolated strains, as confirmed in about the half of cases or probable/possible in the remaining ones, were identified as S. enterica serovar Typhimurium [4,5:i:-], monophasic variant. Aterno river, bordering the L’Aquila district, was recognized as the main responsible source for the contamination of local crops and vegetables derived from polluted crops. Conclusions: The high rate of hospitalized children underlines the emergence of a highly pathogenic S. enterica strain probably subsequent to the contamination of the spring water sources after geological changes occurred during the catastrophic earthquake.

  2. Early Diagnosis and Intervention for Hearing Loss in Newborns Discharged from Intensive Care Units: a Four-year Follow-up Study in North of Iran

    Directory of Open Access Journals (Sweden)

    Sima Tajik

    2016-08-01

    Full Text Available Background Hearing loss is the most common congenital disorder the incidence of which is further increased in the presence of risk factors for hearing loss among newborns admitted to the neonatal intensive care unit (NICU. The aim of this study was early diagnosis and intervention for hearing loss in newborns discharged from NICU. Materials and Methods This prospective cohort study was conducted on 3,362 newborns discharged from the NICU in several hospitals in Babol, Iran. Each newborn was evaluated through the transient evoked otoacoustic emission (T EOAE test. In the absence of any result, retests including TEOAE and diagnostic auditory brainstem response (ABR were conducted. In case of hearing loss, intervention programs including hearing aids fitting and cochlear implant were considered for infants. Each newborn infant was follow-up for four years. The infant’s age was also calculated during the hearing loss diagnosis and the intervention program. Results Sensorineural hearing loss (SNHL was diagnosed in 35 (1.04% of the infants at an average age of 105.65 + 96.72 days. Most of hearing loss diagnosis (51.43% was before the age of 3 months. Hearing aids were fitted for 25 infants (80.64% with a mean age of 9.61 + 7.64 months. Cochlear implants were done for two (8% children. At the end of the follow up, all of the children except one case (3.22% were able to use verbal communication. Conclusion Hearing screening of the high risk NICU graduate babies has reduced the age of hearing loss diagnosis to 3 months. The presence of severe to profound hearing loss in this population highlights the importance of early diagnosis and intervention.

  3. Fuel cell powered by natural gas. The projet `Gazel` four years later; La pile a combustible au gaz naturel. Le projet `Gazel` apres quatre ans de exploitation

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, D.N. [Service de Gaz, Services Industriels de Geneve (Switzerland)

    1997-08-01

    The Services Industriels of Geneve (SIG) acted as a pioneer when in 1993 they purchased and commenced the operation of a first generation phosphoric acid fuel cell ONSI PC25A. The object of the SIG fuel cell pilot project, Gazel, is to promote the technology of fuel cells powered by natural gas, by demonstrating the feasibility of exploitation of the technique, and by seeking to optimize profitability conditions in an installation on an industrial scale. For almost four years, the operational results obtained conform to, and in some cases even exceed, the initial estimates, which were already very favourable in comparison with traditional technologies of electricity production. Indeed, ecological and technical performances more than corresponds to the manufacturer expectations. Long duration tests should allow to evaluate the profitability of the fuel cell, justly considered to be a technology of the future for the gas industry. (orig.) [Deutsch] Im Jahre 1993 haben die Industriellen Betriebe in Genf (SIG) auf eigene Rechnung die Brennstoffzelle PC25A von ONSI erworben und als erste deren regulaeren Betrieb aufgenommen. Mit diesem Projekt hatten die SIG sich zum Ziel gesetzt, die Brennstoffzellen-Technologie zu lancieren. Erdgas wird zur Speisung der Brennstoffzelle benutzt, und das Projekt ist eine Demonstration des zuverlaessigen Betriebs dieser Technologie, mit dem im industriellen Massstab die Rentabilitaetsbedingungen reell optimiert werden. Die in den beinahe vier Jahrzehnten Betriebsdauer erzielten resultate entsprechen voll den Erwartungen und uebertreffen in einzelnen Details die urspruenglichen Vorgaben, die im Vergleich zu den herkoemmlichen Technologien der Elektrizitaetserzeugung bereits sehr viel guenstiger sind. Den an sie gestellten technischen und oekologischen Forderungen von Seiten des Herstellers hat die realisierte Anlage tatsaechlich voellig entsprochen. Langzeitversuche dieser Art sind ideal, um die Rentabilitaet der Brennstoffzelle

  4. Lower Protein-to-Carbohydrate Ratio in Maternal Diet is Associated with Higher Childhood Systolic Blood Pressure up to Age Four Years

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    Michelle L. Blumfield

    2015-04-01

    Full Text Available The prenatal environment can influence development of offspring blood pressure (BP, which tracks into adulthood. This prospective longitudinal study investigated whether maternal pregnancy dietary intake is associated with the development of child BP up to age four years. Data are from 129 mother-child dyads enrolled in the Women and Their Children’s Health study. Maternal diet was assessed using a validated 74-item food frequency questionnaire at 18 to 24 weeks and 36 to 40 weeks, with a reference period of the previous three months. Child systolic and diastolic BP were measured at 3, 6, 9, 12, 24, 36 and 48 months, using an automated BP monitor. Using mixed-model regression analyses adjusted for childhood growth indices, pregnancy intakes of percentage of energy (E% polyunsaturated fat (β coefficient 0.73; 95% CI 0.003, 1.45; p = 0.045, E% omega-6 fatty acids (β coefficient 0.89; 95% CI 0.09, 1.69; p = 0.03 and protein-to-carbohydrate (P:C ratio (β coefficient −14.14; 95% CI −27.68, −0.60; p = 0.04 were associated with child systolic BP trajectory up to 4 years. Child systolic BP was greatest at low proportions of dietary protein (<16% of energy and high carbohydrate (>40% of energy intakes. There may be an ideal maternal macronutrient ratio associated with optimal infant BP. Maternal diet, which is potentially modifiable, may play an important role in influencing offspring risk of future hypertension.

  5. Four-year comparison of water contents beneath a grass ley and a deciduous oak wood overlying Triassic sandstone in lowland England

    Science.gov (United States)

    Green, Julian C.; Reid, Ian; Calder, Ian R.; Nisbet, Thomas R.

    2006-09-01

    SummaryDifferences in the seasonal water dynamics of a sand soil overlying Triassic sandstone have been investigated to a depth of 9 m beneath both a grass ley and a pendunculate oak woodland within Clipstone Forest, Nottinghamshire, England. Fortnightly measurements with a neutron-probe over a four-year period allowed a comparison of the soil-rock water content to depths rarely reported. In spring, the rate of decrease of water content of the uppermost 2 m of soil was much greater under grass than oak woodland. In contrast, the rate of decrease under oak was greater after leafing out in May, while the rate of rewetting in early autumn was lower for this land-use until senescence and leaf-fall in late autumn. In the uppermost 2 m of soil - a depth that includes all plant roots - the soil moisture minima were between 47 and 58 mm lower under oak than grass in each year of monitoring. As a result of both these drier conditions and the comparatively late leaf fall of this species, penetration of the winter-season wetting front to 2 m was delayed by between one and three months at the oak site relative to grass in years of near-average rainfall. Rewetting at 9 m lags by 10-12 months, compared to the surface, giving an average penetration rate for the wetting front to this depth of 25-30 mm day -1, with little observable differences between the land-uses. Preferential flow is evident under both sites, affecting the profile to 3 m in all years and to at least 6 m following winters experiencing exceptional rainfall.

  6. Pre- and postnatal exposure of children to tobacco smoke during the first four years of life – observations of the authors

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    Barbara Kamer

    2014-11-01

    Full Text Available [b]Introduction[/b]. Environmental exposure to tobacco smoke is a significant threat for human health, where the higher is its degree, the more immature the human organism is. Therefore, the exposure to Tobacco smoke in foetal life exerts unfavourable effects on developing foetus and may cause early and distant results in children. [b]Material and methods.[/b] The study comprised 318 children in their first four years of life, treated for various medical conditions. The examined children were divided into two groups, Group 1 – children exposed to Tobacco smoke – and Group 2 – a control group with children from non-smoking families. History data were obtained on the basis of a specially designed questionnaire, used by the doctor in an individual conversation with parent. In each third child from the group 1 cotinine concentration in urine was assayed by the method of high performance liquid chromatography-UV-VIS and the cotinine/creatinine ratio was calculated. [b]Results of stud[/b][b]y[/b]. Results demonstrated environmental exposure to tobacco smoke in 173 children (Group 1. Out of them 31.2% were the children whose mothers had smoked also during pregnancy (Subgroup A. The other 119 children from Group 1 were accounted to Subgroup B, i.e., children, where other household members had been smoking cigarettes. A comparative group comprised 143 children from non-smoking families. The results demonstrated then that 17% of all the examined children were those, exposed to tobacco smoke effects already in their foetal life, predisposing them to prematurity and low birth weight. Moreover, it was observed that the young age and lower education level of their parents, together with worse housing conditions, may suggest a predisposing character and role of the mentioned factors.

  7. The application of POSS nanostructures in cartilage tissue engineering: the chondrocyte response to nanoscale geometry.

    Science.gov (United States)

    Oseni, Adelola O; Butler, Peter E; Seifalian, Alexander M

    2015-11-01

    Despite extensive research into cartilage tissue engineering (CTE), there is still no scaffold ideal for clinical applications. Various synthetic and natural polymers have been investigated in vitro and in vivo, but none have reached widespread clinical use. The authors investigate the potential of POSS-PCU, a synthetic nanocomposite polymer, for use in CTE. POSS-PCU is modified with silsesquioxane nanostructures that improve its biological and physical properties. The ability of POSS-PCU to support the growth of ovine nasoseptal chondrocytes was evaluated against a polymer widely used in CTE, polycaprolactone (PCL). Scaffolds with varied concentrations of the POSS molecule were also synthesized to investigate their effect on chondrocyte growth. Chondrocytes were seeded onto scaffold disks (PCU negative control; POSS-PCU 2%, 4%, 6%, 8%; PCL). Cytocompatibilty was evaluated using cell viability, total DNA, collagen and GAG assays. Chondrocytes cultured on POSS-PCU (2% POSS) scaffolds had significantly higher viability than PCL scaffolds (p  0.05). POSS-PCU (6% and 8% POSS) had improved viability and proliferation over an 18 day culture period compared with 2% and 4% POSS-PCU (p < 0.0001). Increasing the percentage of POSS in the scaffolds increased the size of the pores found in the scaffolds (p < 0.05). POSS-PCU has excellent potential for use in CTE. It supports the growth of chondrocytes in vitro and the POSS modification significantly enhances the growth and proliferation of nasoseptal chondrocytes compared with traditional scaffolds such as PCL.

  8. Ski inhibits TGF-β/phospho-Smad3 signaling and accelerates hypertrophic differentiation in chondrocytes.

    Science.gov (United States)

    Kim, Kyung-Ok; Sampson, Erik R; Maynard, Robert D; O'Keefe, Regis J; Chen, Di; Drissi, Hicham; Rosier, Randy N; Hilton, Matthew J; Zuscik, Michael J

    2012-06-01

    Since transforming growing factor-β (TGF-β)/Smad signaling inhibits chondrocyte maturation, endogenous negative regulators of TGF-β signaling are likely also important regulators of the chondrocyte differentiation process. One such negative regulator, Ski, is an oncoprotein that is known to inhibit TGF-β/Smad3 signaling via its interaction with phospho-Smad3 and recruitment of histone deacetylases (HDACs) to the DNA binding complex. Based on this, we hypothesized that Ski inhibits TGF-β signaling and accelerates maturation in chondrocytes via recruitment of HDACs to transcriptional complexes containing Smads. We tested this hypothesis in chick upper sternal chondrocytes (USCs), where gain and loss of Ski expression experiments were performed. Over-expression of Ski not only reversed the inhibitory effect of TGF-β on the expression of hypertrophic marker genes such as type X collagen (colX) and osteocalcin, it induced these genes basally as well. Conversely, knockdown of Ski by RNA interference led to a reduction of colX and osteocalcin expression under basal conditions. Furthermore, Ski blocked TGF-β induction of cyclinD1 and caused a basal up-regulation of Runx2, consistent with the observed acceleration of hypertrophy. Regarding mechanism, not only does Ski associate with phospho-Smad2 and 3, but its association with phospho-Smad3 is required for recruitment of HDAC4 and 5. Implicating this recruitment of HDACs in the phenotypic effects of Ski in chondrocytes, the HDAC inhibitor SAHA reversed the up-regulation of colX and osteocalcin in Ski over-expressing cells. These results suggest that inhibition of TGF-β signaling by Ski, which involves its association with phospho-Smad3 and recruitment of HDAC4 and 5, leads to accelerated chondrocyte differentiation.

  9. Lidocaine induces ROCK-dependent membrane blebbing and subsequent cell death in rabbit articular chondrocytes.

    Science.gov (United States)

    Maeda, Tsutomu; Toyoda, Futoshi; Imai, Shinji; Tanigawa, Hitoshi; Kumagai, Kousuke; Matsuura, Hiroshi; Matsusue, Yoshitaka

    2016-05-01

    Local anesthetics are administered intraarticularly for pain control in orthopedic clinics and surgeries. Although previous studies have shown that local anesthetics can be toxic to chondrocytes, the underlying cellular mechanisms remain unclear. The present study investigates acute cellular responses associated with lidocaine-induced toxicity to articular chondrocytes. Rabbit articular chondrocytes were exposed to lidocaine and their morphological changes were monitored with live cell microscopy. The viability of chondrocytes was evaluated using a fluorescence based LIVE/DEAD assay. Acute treatment of chondrocytes with lidocaine (3-30 mM) induced spherical protrusions on the cell surface (so called "membrane blebbing") in a time- and concentration-dependent manner. The concentration-response relationship for the lidocaine effect was shifted leftward by elevating extracellular pH, as expected for the non-ionized lidocaine being involved in the bleb formation. ROCK (Rho-kinase) inhibitors Y-27632 and fasudil completely prevented the lidocaine-induced membrane blebbing, suggesting that ROCK activation is required for bleb formation. Caspase-3 levels were unchanged by 10 mM lidocaine (p = 0.325) and a caspase inhibitor z-VAD-fmk did not affect the lidocaine-induced blebbing (p = 0.964). GTP-RhoA levels were significantly increased (p ROCK inhibitors or a myosin-II inhibitor blebbistatin (p ROCK-dependent membrane blebbing and thereby produces a cytotoxic effect on chondrocytes. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:754-762, 2016.

  10. Radiation grafting on natural films

    Science.gov (United States)

    Lacroix, M.; Khan, R.; Senna, M.; Sharmin, N.; Salmieri, S.; Safrany, A.

    2014-01-01

    Different methods of polymer grafting using gamma irradiation are reported in the present study for the preparation of newly functionalized biodegradable films, and some important properties related to their mechanical and barrier properties are described. Biodegradable films composed of zein and poly(vinyl alcohol) (PVA) were gamma-irradiated in presence of different ratios of acrylic acid (AAc) monomer for compatibilization purpose. Resulting grafted films (zein/PVA-g-AAc) had their puncture strength (PS=37-40 N mm-1) and puncture deformation (PD=6.5-9.8 mm) improved for 30% and 50% PVA in blend, with 5% AAc under 20 kGy. Methylcellulose (MC)-based films were irradiated in the presence of 2-hydroxyethyl methacrylate (HEMA) or silane, in order to determine the effect of monomer grafting on the mechanical properties of films. It was found that grafted films (MC-g-HEMA and MC-g-silane) using 35% monomer performed higher mechanical properties with PS values of 282-296 N mm-1 and PD of 5.0-5.5 mm under 10 kGy. Compatibilized polycaprolactone (PCL)/chitosan composites were developed via grafting silane in chitosan films. Resulting trilayer grafted composite film (PCL/chitosan-g-silane/PCL) presented superior tensile strength (TS=22 MPa) via possible improvement of interfacial adhesion (PCL/chitosan) when using 25% silane under 10 kGy. Finally, MC-based films containing crystalline nanocellulose (CNC) as a filling agent were prepared and irradiated in presence of trimethylolpropane trimethacrylate (TMPTMA) as a grafted plasticizer. Grafted films (MC-g-TMPTMA) presented superior mechanical properties with a TS of 47.9 MPa and a tensile modulus (TM) of 1792 MPa, possibly due to high yield formation of radicals to promote TMPTMA grafting during irradiation. The addition of CNC led to an additional improvement of the barrier properties, with a significant 25% reduction of water vapor permeability (WVP) of grafted films.

  11. The Knee Joint Loose Body as a Source of Viable Autologous Human Chondrocytes

    Science.gov (United States)

    Melrose, J.

    2016-01-01

    Loose bodies are fragments of cartilage or bone present in the synovial fluid. In the present study we assessed if loose bodies could be used as a source of autologous human chondrocytes for experimental purposes. Histochemical examination of loose bodies and differential enzymatic digestions were undertaken, the isolated cells were cultured in alginate bead microspheres and immunolocalisations were undertaken for chondrogenic markers such as aggrecan, and type II collagen. Isolated loose body cells had high viability (≥90% viable), expressed chondrogenic markers (aggrecan, type II collagen) but no type I collagen. Loose bodies may be a useful source of autologous chondrocytes of high viability. PMID:27349321

  12. The Results of Fetal Chondrocytes Transplantation in Patients with Rheumatoid Arthritis

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    Natalya Krivoruchko

    2014-12-01

    Full Text Available Introduction. Nowadays anti-inflammatory and immunosuppressive therapy has significantly improved the quality of life and prognosis of rheumatoid arthritis (RA. Nevertheless, there are still many patients with progressive rheumatoid inflammation, resulting in the destruction of joints. Cell therapy seems like a promising direction in rheumatology. The aim of our research was to evaluate the efficacy of fetal chondrocyte transplantation in patients with RA.Methods. We examined 60 patients with rheumatoid arthritis (I - III stages between 20 and 63 years of age. They were divided into 2 groups: the first group underwent the fetal chondrocytes transplantation (n = 40, and the second was a control group who got conservative therapy (n = 20. Donor cells were taken from the chondrogenic layer of the humerus or femur heads and hip condyles of human embryos in gestation for 17-20 weeks. A suspension of fetal chondrocytes injected into affected areas of the articular surfaces under X-ray control. Cell viability was determined before the injection. Efficacy of the therapy was assessed by clinical, instrumental, and laboratory tests. This clinical trial was allowed by The Ministry of Public Health and Ethics Committee. All of our patients gave informed consent for the fetal chondrocytes transplantation.Results. Evaluation of the clinical manifestations of RA in the first group of patients showed 3.7 times decrease in pain and 1.6 times relief of synovitis. Complete reduction of contracture was observed in 82% of patients in the first group. Morphometric changes in X-ray demonstrated inhibition of the destruction in articular cartilage and surfaces of bones after transplantation of fetal chondrocytes. The dynamics of morphological changes in synovium showed 2.5 times reduction of the inflammatory reaction. Transplantation of fetal chondrocytes led to a significant reduction in ESR, CRP, fibrinogen , γ-globulin after a period of 12 months (p < 0

  13. Characteristics of Four-years of GOSAT/TANSO-FTS TIR V1.0 CO2 and CH4 Products

    Science.gov (United States)

    Saitoh, N.; Kimoto, S.; Sugimura, R.; Imasu, R.; Shiomi, K.; Kuze, A.; Kataoka, F.; Knuteson, R. O.; Machida, T.; Sawa, Y.; Matsuda, H.

    2015-12-01

    Greenhouse Gases Observing Satellite (GOSAT) was launched on 23 January 2009, and has continued to make global observations, including both nadir and off-nadir measurements, for more than six years since its launch. The thermal infrared (TIR) band of Thermal and Near-infrared Sensor for Carbon Observation Fourier Transform Spectrometer (TANSO-FTS) on board the GOSAT has observed CO2 and CH4 profiles. We have analyzed the four-year data from 2010 through 2013 of the latest released version of the TIR Level 2 (L2) CO2 and CH4 products (V1.0). Comparisons of the TIR upper atmospheric CO2 product with CO2 data from Comprehensive Observation Network for Trace Gases by Airliner (CONTRAIL) aircraft measurements show that the growth rate estimated from the TIR CO2 data is slightly lower than that from the CONTRAIL data. Overall, the TIR V1.0 CO2 product has better quality in the upper troposphere and lower stratosphere than the a priori judging from comparisons with the collocated aircraft data. In spring and summer, however, the quality of the TIR L2 CO2 products became slightly worse than in the other seasons, especially in the low and northern-mid latitudes. This is because the corresponding a priori had a larger bias and the TIR Level 1B (L1B) radiance spectra might have a larger bias in the spring-summer seasons. Here, we have tested several types of correction methods to modify the L1B spectral bias, and then compared CO2 and CH4 concentrations retrieved after applying spectral bias correction factors with coincident CONTRAIL and HIAPER Pole-to-Pole Observation (HIPPO) aircraft data. The comparison results suggest that the L1B spectral bias correction factor should be changed depending on wavelength. In addition, it should be expressed as a function of on-board internal calibration blackbody temperatures. This is because they are weak season-dependent parameters; they were clearly lower in spring and summer.

  14. Graft stability after endothelial keratoplasty

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    Jovanović Vesna

    2015-01-01

    Full Text Available Bacground/Aim. Techniques for replacing the corneal endothelium have been improved. The host-graft interface is the key to graft adhesion and visual recovery. The aim of this study was to establish graft stability after Descemet stripping with endothelial keratoplasty (DSEK, compare it to the graft stability after endothelial keratoplasty with the intact posterior corneal layers (nDSEK in the rabbit cornea, and to investigate the nature of wound healing. Methods. Adult white rabbits (n = 20 were divided in two experimental groups: ten rabbits underwent monocular DSEK, and ten rabbits underwent endothelial keratoplasty without Descemet stripping (nDSEK. On the second postoperative day a horizontal dislocation of the graft was tried using the Lindstrom roller in each animal. Corneas were processed for the light microscopy study. Results. Rolling the Lindstrom instrument over the corneal surface did not cause horizontal dislocation in any of the operated eyes. In the DSEK group light microscopy revealed the lack of inflammation and fibrosis at the clearly distinctive donor-recipient interface (DRI. Retrocorneal membrane was found in two eyes. In nDSEK group, the host Descemet` s membrane (DM was intact without endothelial cells, with good graft apposition, without inflammation, fibrosis, or retrocorneal membrane. Conclusion. This study suggests that there is no difference in graft stability in DSEK compared to nDSEK in rabbit corneas. Wounds healed at DRI by hypocellular scarring only in both experimental groups.

  15. Design of a Sapling Branch Grafting Robot

    Directory of Open Access Journals (Sweden)

    Qun Sun

    2014-01-01

    Full Text Available The automatic sapling grafting methods and grafting robot technologies are not comprehensively studied despite the fact that they are urgently required in practice. For this reason, a sapling grafting robot is developed to implement automatic grafting for saplings. The developed grafting robot includes clipping mechanism, moving mechanism, cutting mechanism, binding mechanism, and Arduino MCU based control system, which is capable of clipping, moving, positioning, cutting, grafting, and binding saplings. Experiments show that the stock cutting efficiency is 98.4%, the scion cutting efficiency is 98.9%, the grafting efficiency is 87.3%, and the binding efficiency is 68.9%.

  16. Growth factor priming differentially modulates components of the extracellular matrix proteome in chondrocytes and synovium-derived stem cells.

    Directory of Open Access Journals (Sweden)

    Elena Alegre-Aguarón

    Full Text Available To make progress in cartilage repair it is essential to optimize protocols for two-dimensional cell expansion. Chondrocytes and SDSCs are promising cell sources for cartilage repair. We previously observed that priming with a specific growth factor cocktail (1 ng/mL transforming growth factor-β1, 5 ng/mL basic fibroblast growth factor, and 10 ng/mL platelet-derived growth factor-BB in two-dimensional culture, led to significant improvement in mechanical and biochemical properties of synovium-derived stem cell (SDSC-seeded constructs. The current study assessed the effect of growth factor priming on the proteome of canine chondrocytes and SDSCs. In particular, growth factor priming modulated the proteins associated with the extracellular matrix in two-dimensional cultures of chondrocytes and SDSCs, inducing a partial dedifferentiation of chondrocytes (most proteins associated with cartilage were down-regulated in primed chondrocytes and a partial differentiation of SDSCs (some collagen-related proteins were up-regulated in primed SDSCs. However, when chondrocytes and SDSCs were grown in pellet culture, growth factor-primed cells maintained their chondrogenic potential with respect to glycosaminoglycan and collagen production. In conclusion, the strength of the label-free proteomics technique is that it allows for the determination of changes in components of the extracellular matrix proteome in chondrocytes and SDSCs in response to growth factor priming, which could help in future tissue engineering strategies.

  17. Evidence for lysosomal exocytosis and release of aggrecan-degrading hydrolases from hypertrophic chondrocytes, in vitro and in vivo

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    Edward R. Bastow

    2012-02-01

    The abundant proteoglycan, aggrecan, is resorbed from growth plate cartilage during endochondral bone ossification, yet mice with genetically-ablated aggrecan-degrading activity have no defects in bone formation. To account for this apparent anomaly, we propose that lysosomal hydrolases degrade extracellular, hyaluronan-bound aggrecan aggregates in growth plate cartilage, and that lysosomal hydrolases are released from hypertrophic chondrocytes into growth plate cartilage via Ca2+-dependent lysosomal exocytosis. In this study we confirm that hypertrophic chondrocytes release hydrolases via lysosomal exocytosis in vitro and we show in vivo evidence for lysosomal exocytosis in hypertrophic chondrocytes during skeletal development. We show that lysosome-associated membrane protein 1 (LAMP1 is detected at the cell surface following in vitro treatment of epiphyseal chondrocytes with the calcium ionophore, ionomycin. Furthermore, we show that in addition to the lysosomal exocytosis markers, cathepsin D and β-hexosaminidase, ionomycin induces release of aggrecan- and hyaluronan-degrading activity from cultured epiphyseal chondrocytes. We identify VAMP-8 and VAMP7 as v-SNARE proteins with potential roles in lysosomal exocytosis in hypertrophic chondrocytes, based on their colocalisation with LAMP1 at the cell surface in secondary ossification centers in mouse tibiae. We propose that resorbing growth plate cartilage involves release of destructive hydrolases from hypertrophic chondrocytes, via lysosomal exocytosis.

  18. Zfp521 is a target gene and key effector of parathyroid hormone-related peptide signaling in growth plate chondrocytes.

    Science.gov (United States)

    Correa, Diego; Hesse, Eric; Seriwatanachai, Dutmanee; Kiviranta, Riku; Saito, Hiroaki; Yamana, Kei; Neff, Lynn; Atfi, Azeddine; Coillard, Lucie; Sitara, Despina; Maeda, Yukiko; Warming, Soren; Jenkins, Nancy A; Copeland, Neal G; Horne, William C; Lanske, Beate; Baron, Roland

    2010-10-19

    In the growth plate, the interplay between parathyroid hormone-related peptide (PTHrP) and Indian hedgehog (Ihh) signaling tightly regulates chondrocyte proliferation and differentiation during longitudinal bone growth. We found that PTHrP increases the expression of Zfp521, a zinc finger transcriptional coregulator, in prehypertrophic chondrocytes. Mice with chondrocyte-targeted deletion of Zfp521 resembled PTHrP(-/-) and chondrocyte-specific PTHR1(-/-) mice, with decreased chondrocyte proliferation, early hypertrophic transition, and reduced growth plate thickness. Deleting Zfp521 increased expression of Runx2 and Runx2 target genes, and decreased Cyclin D1 and Bcl-2 expression while increasing Caspase-3 activation and apoptosis. Zfp521 associated with Runx2 in chondrocytes, antagonizing its activity via an HDAC4-dependent mechanism. PTHrP failed to upregulate Cyclin D1 and to antagonize Runx2, Ihh, and collagen X expression when Zfp521 was absent. Thus, Zfp521 is an important PTHrP target gene that regulates growth plate chondrocyte proliferation and differentiation.

  19. Chondrogenic differentiation of human articular chondrocytes differs in biodegradable PGA/PLA scaffolds

    DEFF Research Database (Denmark)

    Zwingmann, Joern; Mehlhorn, Alexander T; Südkamp, Norbert

    2007-01-01

    Cartilage tissue engineering is applied clinically to cover and regenerate articular cartilage defects. Two bioresorbable nonwoven scaffolds, polyglycolic acid (PGA) and poly(lactic-co-glycolic acid) (PLGA) (90/10 copolymer of L-lactide and glycolide), were seeded with human chondrocytes after in...

  20. MRTF-A signaling regulates the acquisition of the contractile phenotype in dedifferentiated chondrocytes.

    Science.gov (United States)

    Parreno, Justin; Raju, Sneha; Wu, Po-Han; Kandel, Rita A

    2016-10-14

    Chondrocyte culture as a monolayer for cell number expansion results in dedifferentiation whereby expanded cells acquire contractile features and increased actin polymerization status. This study determined whether the actin polymerization based signaling pathway, myocardin-related transcription factor-a (MRTF-A) is involved in regulating this contractile phenotype. Serial passaging of chondrocytes in monolayer culture to passage 2 resulted in increased gene and protein expression of the contractile molecules alpha-smooth muscle actin, transgelin and vinculin compared to non-passaged, primary cells. This resulted in a functional change as passaged 2, but not primary, chondrocytes were capable of contracting type I collagen gels in a stress-relaxed contraction assay. These changes were associated with increased actin polymerization and MRTF-A nuclear localization. The involvement of actin was demonstrated by latrunculin B depolymerization of actin which reversed these changes. Alternatively cytochalasin D which activates MRTF-A increased gene and protein expression of α-smooth muscle actin, transgelin and vinculin, whereas CCG1423 which deactivates MRTF-A decreased these molecules. The involvement of MRTF-A signaling was confirmed by gene silencing of MRTF or its co-factor serum response factor. Knockdown experiments revealed downregulation of α-smooth muscle actin and transgelin gene and protein expression, and inhibition of gel contraction. These findings demonstrate that passaged chondrocytes acquire a contractile phenotype and that this change is modulated by the actin-MRTF-A-serum response factor signaling pathway.

  1. 3D Culture of Chondrocytes in Gelatin Hydrogels with Different Stiffness

    Directory of Open Access Journals (Sweden)

    Xiaomeng Li

    2016-07-01

    Full Text Available Gelatin hydrogels can mimic the microenvironments of natural tissues and encapsulate cells homogeneously, which makes them attractive for cartilage tissue engineering. Both the mechanical and biochemical properties of hydrogels can affect the phenotype of chondrocytes. However, the influence of each property on chondrocyte phenotype is unclear due to the difficulty in separating the roles of these properties. In this study, we aimed to study the influence of hydrogel stiffness on chondrocyte phenotype while excluding the role of biochemical factors, such as adhesion site density in the hydrogels. By altering the degree of methacryloyl functionalization, gelatin hydrogels with different stiffnesses of 3.8, 17.1, and 29.9 kPa Young’s modulus were prepared from the same concentration of gelatin methacryloyl (GelMA macromers. Bovine articular chondrocytes were encapsulated in the hydrogels and cultured for 14 days. The influence of hydrogel stiffness on the cell behaviors including cell viability, cell morphology, and maintenance of chondrogenic phenotype was evaluated. GelMA hydrogels with high stiffness (29.9 kPa showed the best results on maintaining chondrogenic phenotype. These results will be useful for the design and preparation of scaffolds for cartilage tissue engineering.

  2. Acquiring Chondrocyte Phenotype from Human Mesenchymal Stem Cells under Inflammatory Conditions

    Directory of Open Access Journals (Sweden)

    Masahiro Kondo

    2014-11-01

    Full Text Available An inflammatory milieu breaks down the cartilage matrix and induces chondrocyte apoptosis, resulting in cartilage destruction in patients with cartilage degenerative diseases, such as rheumatoid arthritis or osteoarthritis. Because of the limited regenerative ability of chondrocytes, defects in cartilage are irreversible and difficult to repair. Mesenchymal stem cells (MSCs are expected to be a new tool for cartilage repair because they are present in the cartilage and are able to differentiate into multiple lineages of cells, including chondrocytes. Although clinical trials using MSCs for patients with cartilage defects have already begun, its efficacy and repair mechanisms remain unknown. A PubMed search conducted in October 2014 using the following medical subject headings (MeSH terms: mesenchymal stromal cells, chondrogenesis, and cytokines resulted in 204 articles. The titles and abstracts were screened and nine articles relevant to “inflammatory” cytokines and “human” MSCs were identified. Herein, we review the cell biology and mechanisms of chondrocyte phenotype acquisition from human MSCs in an inflammatory milieu and discuss the clinical potential of MSCs for cartilage repair.

  3. Beta1 integrins regulate chondrocyte rotation, G1 progression, and cytokinesis

    DEFF Research Database (Denmark)

    Aszodi, Attila; Hunziker, Ernst B; Brakebusch, Cord;

    2003-01-01

    -actin organization. In addition, mutant chondrocytes show decreased proliferation caused by a defect in G1/S transition and cytokinesis. The G1/S defect is, at least partially, caused by overexpression of Fgfr3, nuclear translocation of Stat1/Stat5a, and up-regulation of the cell cycle inhibitors p16 and p21...

  4. Upregulation of matrix synthesis in chondrocyte-seeded agarose following sustained bi-axial cyclic loading

    Directory of Open Access Journals (Sweden)

    Belinda Pingguan-Murphy

    2012-08-01

    Full Text Available OBJECTIVES: The promotion of extracellular matrix synthesis by chondrocytes is a requisite part of an effective cartilage tissue engineering strategy. The aim of this in vitro study was to determine the effect of bi-axial cyclic mechanical loading on cell proliferation and the synthesis of glycosaminoglycans by chondrocytes in threedimensional cultures. METHOD: A strain comprising 10% direct compression and 1% compressive shear was applied to bovine chondrocytes seeded in an agarose gel during two 12-hour conditioning periods separated by a 12-hour resting period. RESULTS: The bi-axial-loaded chondrocytes demonstrated a significant increase in glycosaminoglycan synthesis compared with samples exposed to uni-axial or no loading over the same period (p<0.05. The use of a free-swelling recovery period prior to the loading regime resulted in additional glycosaminoglycan production and a significant increase in DNA content (p<0.05, indicating cell proliferation. CONCLUSIONS: These results demonstrate that the use of a bi-axial loading regime results in increased matrix production compared with uni-axial loading.

  5. Polyethylene-glycol-modified single-walled carbon nanotubes for intra-articular delivery to chondrocytes.

    Science.gov (United States)

    Sacchetti, Cristiano; Liu-Bryan, Ru; Magrini, Andrea; Rosato, Nicola; Bottini, Nunzio; Bottini, Massimo

    2014-12-23

    Osteoarthritis (OA) is a common and debilitating degenerative disease of articular joints for which no disease-modifying medical therapy is currently available. Inefficient delivery of pharmacologic agents into cartilage-resident chondrocytes after systemic administration has been a limitation to the development of anti-OA medications. Direct intra-articular injection enables delivery of high concentrations of agents in close proximity to chondrocytes; however, the efficacy of this approach is limited by the fast clearance of small molecules and biomacromolecules after injection into the synovial cavity. Coupling of pharmacologic agents with drug delivery systems able to enhance their residence time and cartilage penetration can enhance the effectiveness of intra-articularly injected anti-OA medications. Herein we describe an efficient intra-articular delivery nanosystem based on single-walled carbon nanotubes (SWCNTs) modified with polyethylene glycol (PEG) chains (PEG-SWCNTs). We show that PEG-SWCNTs are capable to persist in the joint cavity for a prolonged time, enter the cartilage matrix, and deliver gene inhibitors into chondrocytes of both healthy and OA mice. PEG-SWCNT nanoparticles did not elicit systemic or local side effects. Our data suggest that PEG-SWCNTs represent a biocompatible and effective nanocarrier for intra-articular delivery of agents to chondrocytes.

  6. A new test method for the standardized evaluation of changes in the ultrastructure of chondrocytes.

    Science.gov (United States)

    Annefeld, M

    1985-01-01

    By means of a new ultrastructural test system, which is based on standardized morphometry and statistical evaluation, we are able for the first time to quantify changes in the cell metabolism of chondrocytes in the articular cartilage. To compare anti-inflammatory substances of different structure, different absorption characteristics and pharmacokinetics as regards their effect on rat cartilage, we used an equieffective dosage of the different anti-inflammatory drugs. Dexamethasone as steroidal and indomethacin and phenylbutazone as classical non-steroidal anti-inflammatory drugs (NSAIDs) were administered in an ED50 dosage referred to provoked arthritis over 12 weeks, using the same mode of administration. The standardized results of untreated rats weighing 300 and 450 g were used as the controls. Dexamethasone brings about massive degenerative changes in the ultrastructure of the vital chondrocyte. Under indomethacin and phenylbutazone the metabolic activity of the chondrocyte is inhibited to a much lesser extent. The damage to the chondrocyte after treatment with dexamethasone, indomethacin and phenylbutazone cannot be regarded as minimal but in some cases is tolerable as regards the benefit/risk ratio in the treatment of rheumatoid diseases.

  7. Chondrogenic potential of articular chondrocytes depends on their original location in the knee

    NARCIS (Netherlands)

    Bekkers, J.E.J.; Saris, D.B.F.; Tsuchida, A.I.; Rijen, van M.H.P.; Dhert, W.J.A.; Creemers, L.

    2014-01-01

    Objective: This study aimed to investigate the regenerative capacity of chondrocytes derived from debrided defect cartilage and healthy cartilage from different regions in the joint in order to determine the best cell source for regenerative cartilage therapies. Methods: Articular cartilage was obta

  8. Profilin 1 is required for abscission during late cytokinesis of chondrocytes

    Science.gov (United States)

    Böttcher, Ralph T; Wiesner, Sebastian; Braun, Attila; Wimmer, Reiner; Berna, Alejandro; Elad, Nadav; Medalia, Ohad; Pfeifer, Alexander; Aszódi, Attila; Costell, Mercedes; Fässler, Reinhard

    2009-01-01

    Profilins are key factors for dynamic rearrangements of the actin cytoskeleton. However, the functions of profilins in differentiated mammalian cells are uncertain because profilin deficiency is early embryonic lethal for higher eukaryotes. To examine profilin function in chondrocytes, we disrupted the profilin 1 gene in cartilage (Col2pfn1). Homozygous Col2pfn1 mice develop progressive chondrodysplasia caused by disorganization of the growth plate and defective chondrocyte cytokinesis, indicated by the appearance of binucleated cells. Surprisingly, Col2pfn1 chondrocytes assemble and contract actomyosin rings normally during cell division; however, they display defects during late cytokinesis as they frequently fail to complete abscission due to their inability to develop strong traction forces. This reduced force generation results from an impaired formation of lamellipodia, focal adhesions and stress fibres, which in part could be linked to an impaired mDia1-mediated actin filament elongation. Neither an actin nor a poly-proline binding-deficient profilin 1 is able to rescue the defects. Taken together, our results demonstrate that profilin 1 is not required for actomyosin ring formation in dividing chondrocytes but necessary to generate sufficient force for abscission during late cytokinesis. PMID:19262563

  9. Role of lubricin and boundary lubrication in the prevention of chondrocyte apoptosis.

    Science.gov (United States)

    Waller, Kimberly A; Zhang, Ling X; Elsaid, Khaled A; Fleming, Braden C; Warman, Matthew L; Jay, Gregory D

    2013-04-01

    Osteoarthritis is a complex disease involving the mechanical breakdown of articular cartilage in the presence of altered joint mechanics and chondrocyte death, but the connection between these factors is not well established. Lubricin, a mucinous glycoprotein encoded by the PRG4 gene, provides boundary lubrication in articular joints. Joint friction is elevated and accompanied by accelerated cartilage damage in humans and mice that have genetic deficiency of lubricin. Here, we investigated the relationship between coefficient of friction and chondrocyte death using ex vivo and in vitro measurements of friction and apoptosis. We observed increases in whole-joint friction and cellular apoptosis in lubricin knockout mice compared with wild-type mice. When we used an in vitro bovine explant cartilage-on-cartilage bearing system, we observed a direct correlation between coefficient of friction and chondrocyte apoptosis in the superficial layers of cartilage. In the bovine explant system, the addition of lubricin as a test lubricant significantly lowered the static coefficient of friction and number of apoptotic chondrocytes. These results demonstrate a direct connection between lubricin, boundary lubrication, and cell survival and suggest that supplementation of synovial fluid with lubricin may be an effective treatment to prevent cartilage deterioration in patients with genetic or acquired deficiency of lubricin.

  10. Evaluation of a mPEG-polyester-based hydrogel as cell carrier for chondrocytes.

    Science.gov (United States)

    Peng, Sydney; Yang, Shu-Rui; Ko, Chao-Yin; Peng, Yu-Shiang; Chu, I-Ming

    2013-11-01

    Temperature-sensitive hydrogels are attractive alternatives to porous cell-seeded scaffolds and is minimally invasive through simple injection and in situ gelling. In this study, we compared the performance of two types of temperature-sensitive hydrogels on chondrocytes encapsulation for the use of tissue engineering of cartilage. The two hydrogels are composed of methoxy poly(ethylene glycol)- poly(lactic-co-valerolactone) (mPEG-PVLA), and methoxy poly(ethylene glycol)-poly(lactic- co-glycolide) (mPEG-PLGA). Osmolarity and pH were optimized through the manipulation of polymer concentration and dispersion medium. Chondrocytes proliferation in mPEG-PVLA hydrogels was observed as well as accumulation of GAGs and collagen. On the other hand, chondrocytes encapsulated in mPEG-PLGA hydrogels showed low viability and chondrogenesis. Also, mPEG-PVLA hydrogel, which is more hydrophobic, retained physical integrity after 14 days while mPEG-PLGA hydrogel underwent full degradation due to faster hydrolysis rate and more pronounced acidic self-catalyzed degradation. The mPEG-PVLA hydrogel can be furthered tuned by manipulation of molecular weights to obtain hydrogels with different swelling and degradation characteristics, which may be useful as producing a selection of hydrogels compatible with different cell types. Taken together, these results demonstrate that mPEG-PVLA hydrogels are promising to serve as three-dimensional cell carriers for chondrocytes and potentially applicable in cartilage tissue engineering.

  11. Oxidative Stress Promotes Peroxiredoxin Hyperoxidation and Attenuates Pro-survival Signaling in Aging Chondrocytes.

    Science.gov (United States)

    Collins, John A; Wood, Scott T; Nelson, Kimberly J; Rowe, Meredith A; Carlson, Cathy S; Chubinskaya, Susan; Poole, Leslie B; Furdui, Cristina M; Loeser, Richard F

    2016-03-25

    Oxidative stress-mediated post-translational modifications of redox-sensitive proteins are postulated as a key mechanism underlying age-related cellular dysfunction and disease progression. Peroxiredoxins (PRX) are critical intracellular antioxidants that also regulate redox signaling events. Age-related osteoarthritis is a common form of arthritis that has been associated with mitochondrial dysfunction and oxidative stress. The objective of this study was to determine the effect of aging and oxidative stress on chondrocyte intracellular signaling, with a specific focus on oxidation of cytosolic PRX2 and mitochondrial PRX3. Menadione was used as a model to induce cellular oxidative stress. Compared with chondrocytes isolated from young adult humans, chondrocytes from older adults exhibited higher levels of PRX1-3 hyperoxidation basally and under conditions of oxidative stress. Peroxiredoxin hyperoxidation was associated with inhibition of pro-survival Akt signaling and stimulation of pro-death p38 signaling. These changes were prevented in cultured human chondrocytes by adenoviral expression of catalase targeted to the mitochondria (MCAT) and in cartilage explants from MCAT transgenic mice. Peroxiredoxin hyperoxidation was observedin situin human cartilage sections from older adults and in osteoarthritic cartilage. MCAT transgenic mice exhibited less age-related osteoarthritis. These findings demonstrate that age-related oxidative stress can disrupt normal physiological signaling and contribute to osteoarthritis and suggest peroxiredoxin hyperoxidation as a potential mechanism.

  12. Chondrocyte BMP2 signaling plays an essential role in bone fracture healing.

    Science.gov (United States)

    Mi, Meng; Jin, Hongting; Wang, Baoli; Yukata, Kiminori; Sheu, Tzong-Jen; Ke, Qiao Han; Tong, Peijian; Im, Hee-Jeong; Xiao, Guozhi; Chen, Di

    2013-01-10

    The specific role of endogenous Bmp2 gene in chondrocytes and in osteoblasts in fracture healing was investigated by generation and analysis of chondrocyte- and osteoblast-specific Bmp2 conditional knockout (cKO) mice. The unilateral open transverse tibial fractures were created in these Bmp2 cKO mice. Bone fracture callus samples were collected and analyzed by X-ray, micro-CT, histology analyses, biomechanical testing and gene expression assays. The results demonstrated that the lack of Bmp2 expression in chondrocytes leads to a prolonged cartilage callus formation and a delayed osteogenesis initiation and progression into mineralization phase with lower biomechanical properties. In contrast, when the Bmp2 gene was deleted in osteoblasts, the mice showed no significant difference in the fracture healing process compared to control mice. These findings suggest that endogenous BMP2 expression in chondrocytes may play an essential role in cartilage callus maturation at an early stage of fracture healing. Our studies may provide important information for clinical application of BMP2.

  13. Repairing cartilage defects using chondrocyte and osteoblast composites developed using a bioreactor

    Institute of Scientific and Technical Information of China (English)

    SUN Shui; REN Qiang; WANG Dong; ZHANG Lei; WU Shuai; SUN Xi-tao

    2011-01-01

    Background Articular cartilage injury is a common disease, and the incidence of articular wear, degeneration, trauma and sports injury is increasing, which often lead to disability and reduced quality of life. Unfortunately repair of articular cartilage defects do not always provide satisfactory outcomes.Methods Chondrocyte and osteoblast composites were co-cultured using a bioreactor. The cartilage defects were treated with cell-β-tricalcium phosphate (β-TCP) composites implanted into osteochondral defects in dogs, in vivo, using mosaicplasty, by placing chondrocyte-β-TCP scaffold composites on top of the defect and osteoblast-β-TCP scaffold composites below the defect.Results Electron microscopy revealed that the induced chondrocytes and osteoblast showed fine adhesive progression and proliferation in the β-TCP scaffold. The repaired tissues in the experimental group maintained their thickness to the full depth of the original defects, as compared with the negative control group (q=12.3370, P <0.01; q=31.5393, P <0.01).Conclusions Perfusion culture provided sustained nutrient supply and gas exchange into the center of the large scaffold. This perfusion bioreactor enables the chondrocytes and osteoblasts to survive and proliferate in a three-dimensional scaffold.

  14. The effect of compressive loading magnitude on in situ chondrocyte calcium signaling.

    Science.gov (United States)

    Madden, Ryan M J; Han, Sang-Kuy; Herzog, Walter

    2015-01-01

    Chondrocyte metabolism is stimulated by deformation and is associated with structural changes in the cartilage extracellular matrix (ECM), suggesting that these cells are involved in maintaining tissue health and integrity. Calcium signaling is an initial step in chondrocyte mechanotransduction that has been linked to many cellular processes. Previous studies using isolated chondrocytes proposed loading magnitude as an important factor regulating this response. However, calcium signaling in the intact cartilage differs compared to isolated cells. The purpose of this study was to investigate the effect of loading magnitude on chondrocyte calcium signaling in intact cartilage. We hypothesized that the percentage of cells exhibiting at least one calcium signal increases with increasing load. Fully intact rabbit femoral condyle and patellar bone/cartilage samples were incubated in calcium-sensitive dyes and imaged continuously under compressive loads of 10-40 % strain. Calcium signaling was primarily associated with the dynamic loading phase and greatly increased beyond a threshold deformation of about 10 % nominal tissue strain. There was a trend toward more cells exhibiting calcium signaling as loading magnitude increased (p = 0.133). These results provide novel information toward identifying mechanisms underlying calcium-dependent signaling pathways related to cartilage homeostasis and possibly the onset and progression of osteoarthritis.

  15. Influence of cell quality on clinical outcome after autologous chondrocyte implantation

    DEFF Research Database (Denmark)

    Niemeyer, Philipp; Pestka, Jan M; Salzmann, Gian M;

    2012-01-01

    BACKGROUND: Several factors influence clinical outcome after autologous chondrocyte implantation (ACI) for the treatment of cartilage defects of the knee joint. HYPOTHESIS/PURPOSE: The aim of the present study was to investigate the influence of cell quality on clinical outcome after ACI. The hyp...

  16. Clinical outcomes after cell-seeded autologous chondrocyte implantation of the knee

    DEFF Research Database (Denmark)

    Pestka, Jan M; Bode, Gerrit; Salzmann, Gian;

    2014-01-01

    BACKGROUND: Autologous chondrocyte implantation (ACI) has been associated with satisfying results. Still, it remains unclear when success or failure after ACI can be estimated. PURPOSE: To evaluate the clinical outcomes of cell-seeded collagen matrix-supported ACI (ACI-Cs) for the treatment of ca...

  17. Effect of Collagen Type I or Type II on Chondrogenesis by Cultured Human Articular Chondrocytes

    NARCIS (Netherlands)

    Rutgers, M.; Saris, D.B.F.; Vonk, L.A.; Rijen, van M.H.P.; Akrum, V.; Langeveld, D.; Boxtel, van A.; Dhert, W.J.A.; Creemers, L.B.

    2013-01-01

    Introduction: Current cartilage repair procedures using autologous chondrocytes rely on a variety of carriers for implantation. Collagen types I and II are frequently used and valuable properties of both were shown earlier in vitro, although a preference for either was not demonstrated. Recently, ho

  18. Surviving endoplasmic reticulum stress is coupled to altered chondrocyte differentiation and function.

    Directory of Open Access Journals (Sweden)

    Kwok Yeung Tsang

    2007-03-01

    Full Text Available In protein folding and secretion disorders, activation of endoplasmic reticulum (ER stress signaling (ERSS protects cells, alleviating stress that would otherwise trigger apoptosis. Whether the stress-surviving cells resume normal function is not known. We studied the in vivo impact of ER stress in terminally differentiating hypertrophic chondrocytes (HCs during endochondral bone formation. In transgenic mice expressing mutant collagen X as a consequence of a 13-base pair deletion in Col10a1 (13del, misfolded alpha1(X chains accumulate in HCs and elicit ERSS. Histological and gene expression analyses showed that these chondrocytes survived ER stress, but terminal differentiation is interrupted, and endochondral bone formation is delayed, producing a chondrodysplasia phenotype. This altered differentiation involves cell-cycle re-entry, the re-expression of genes characteristic of a prehypertrophic-like state, and is cell-autonomous. Concomitantly, expression of Col10a1 and 13del mRNAs are reduced, and ER stress is alleviated. ERSS, abnormal chondrocyte differentiation, and altered growth plate architecture also occur in mice expressing mutant collagen II and aggrecan. Alteration of the differentiation program in chondrocytes expressing unfolded or misfolded proteins may be part of an adaptive response that facilitates survival and recovery from the ensuing ER stress. However, the altered differentiation disrupts the highly coordinated events of endochondral ossification culminating in chondrodysplasia.

  19. Induction of vascular endothelial growth factor by nitric oxide in cultured human articular chondrocytes.

    Science.gov (United States)

    Turpaev, K; Litvinov, D; Dubovaya, V; Panasyuk, A; Ivanov, D; Prassolov, V

    2001-06-01

    We investigated the role of nitric oxide (NO) in the control of vascular endothelial growth factor A (VEGF) gene expression in cultured human articular chondrocytes. Cell treatment with the NO-generating compound nitrosoglutathione (GSNO) caused a significant accumulation of 4.4 kb VEGF mRNA, a major VEGF mRNA isoform expressing in chondrocytes. This is the first demonstration that NO can induce VEGF mRNA expression in chondrocytes. VEGF mRNA level was not affected in cells exposed to dibutyryl cGMP, a non-hydrolyzable analog of cGMP, suggesting that the cGMP system is not involved in NO-dependent transcriptional activation of VEGF gene. The GSNO-stimulated induction of VEGF mRNA was slightly attenuated by MAP protein kinase inhibitors PD98058 and SB203580, but was completely blocked in cells incubated with GSNO in the presence of catalase and superoxide dismutase, enzymes scavenging reactive oxygen species (ROS), or in the presence of thiol-containing antioxidants, N-acetyl cysteine and reduced glutathione. These results suggest that in articular chondrocytes the GSNO-induced VEGF gene transcriptional activation is dependent on endogenous ROS production and oxidative thiol modifications.

  20. Increased adipogenesis in cultured embryonic chondrocytes and in adult bone marrow of dominant negative Erg transgenic mice.

    Directory of Open Access Journals (Sweden)

    Sébastien Flajollet

    Full Text Available In monolayer culture, primary articular chondrocytes have an intrinsic tendency to lose their phenotype during expansion. The molecular events underlying this chondrocyte dedifferentiation are still largely unknown. Several transcription factors are important for chondrocyte differentiation. The Ets transcription factor family may be involved in skeletal development. One family member, the Erg gene, is mainly expressed during cartilage formation. To further investigate the potential role of Erg in the maintenance of the chondrocyte phenotype, we isolated and cultured chondrocytes from the rib cartilage of embryos of transgenic mice that express a dominant negative form of Erg (DN-Erg during cartilage formation. DN-Erg expression in chondrocytes cultured for up to 20 days did not affect the early dedifferentiation usually observed in cultured chondrocytes. However, lipid droplets accumulated in DN-Erg chondrocytes, suggesting adipocyte emergence. Transcriptomic analysis using a DNA microarray, validated by quantitative RT-PCR, revealed strong differential gene expression, with a decrease in chondrogenesis-related markers and an increase in adipogenesis-related gene expression in cultured DN-Erg chondrocytes. These results indicate that Erg is involved in either maintaining the chondrogenic phenotype in vitro or in cell fate orientation. Along with the in vitro studies, we compared adipocyte presence in wild-type and transgenic mice skeletons. Histological investigations revealed an increase in the number of adipocytes in the bone marrow of adult DN-Erg mice even though no adipocytes were detected in embryonic cartilage or bone. These findings suggest that the Ets transcription factor family may contribute to the homeostatic balance in skeleton cell plasticity.

  1. Culture temperature affects human chondrocyte messenger RNA expression in monolayer and pellet culture systems.

    Directory of Open Access Journals (Sweden)

    Akira Ito

    Full Text Available Cell-based therapy has been explored for articular cartilage regeneration. Autologous chondrocyte implantation is a promising cell-based technique for repairing articular cartilage defects. However, there are several issues such as chondrocyte de-differentiation. While numerous studies have been designed to overcome some of these issues, only a few have focused on the thermal environment that can affect chondrocyte metabolism and phenotype. In this study, the effects of different culture temperatures on human chondrocyte metabolism- and phenotype-related gene expression were investigated in 2D and 3D environments. Human chondrocytes were cultured in a monolayer or in a pellet culture system at three different culture temperatures (32°C, 37°C, and 41°C for 3 days. The results showed that the total RNA level, normalized to the threshold cycle value of internal reference genes, was higher at lower temperatures in both culture systems. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH and citrate synthase (CS, which are involved in glycolysis and the citric acid cycle, respectively, were expressed at similar levels at 32°C and 37°C in pellet cultures, but the levels were significantly lower at 41°C. Expression of the chondrogenic markers, collagen type IIA1 (COL2A1 and aggrecan (ACAN, was higher at 37°C than at 32°C and 41°C in both culture systems. However, this phenomenon did not coincide with SRY (sex-determining region Y-box 9 (SOX9, which is a fundamental transcription factor for chondrogenesis, indicating that a SOX9-independent pathway might be involved in this phenomenon. In conclusion, the expression of chondrocyte metabolism-related genes at 32°C was maintained or enhanced compared to that at 37°C. However, chondrogenesis-related genes were further induced at 37°C in both culture systems. Therefore, manipulating the culture temperature may be an advantageous approach for regulating human chondrocyte metabolic activity and

  2. Vascular graft infections with Mycoplasma

    DEFF Research Database (Denmark)

    Levi-Mazloum, Niels Donald; Skov Jensen, J; Prag, J;

    1995-01-01

    Vascular graft infection is one of the most serious complications in vascular surgery. It is associated with mortality rates ranging from 25% to 75% and with morbidity in the form of amputation in approximately 30% of patients. Staphylococcus aureus is the leading pathogen. With conventional...... laboratory techniques, the percentage of culture-negative yet grossly infected vascular grafts seems to be increasing and is not adequately explained by the prior use of antibiotics. We have recently reported the first case of aortic graft infection with Mycoplasma. We therefore suggest the hypothesis...... that the large number of culture-negative yet grossly infected vascular grafts may be due to Mycoplasma infection not detected with conventional laboratory technique....

  3. Interleukin-1 inhibits osmotically-induced calcium signaling and volume regulation in articular chondrocytes

    Science.gov (United States)

    Pritchard, Scott; Votta, Bartholomew J.; Kumar, Sanjay; Guilak, Farshid

    2011-01-01

    OBJECTIVE Articular chondrocytes respond to osmotic stress with transient changes in cell volume and the intracellular concentration of calcium ion ([Ca2+]i). The goal of this study was to examine the hypothesis that interleukin-1 (IL-1), a pro-inflammatory cytokine associated with osteoarthritis, influences osmotically-induced Ca2+ signaling. METHODS Fluorescence ratio imaging was used to measure [Ca2+]i and cell volume in response to hypo- or hyper-osmotic stress in isolated porcine chondrocytes, with or without pre-exposure to 10 ng/ml IL-1α. Inhibitors of IL-1 (IL-1 receptor antagonist, IL-Ra), Ca2+ mobilization (thapsigargin, an inhibitor of Ca-ATPases), and cytoskeletal remodeling (Toxin B, an inhibitor of the Rho family of small GTPases) were used to determine the mechanisms involved in increased [Ca2+]i, F-actin remodeling, volume adaptation and active volume recovery. RESULTS In response to osmotic stress, chondrocytes exhibited transient increases in [Ca2+]i, generally followed by decaying oscillations. Pre-exposure to IL-1 significantly inhibited regulatory volume decrease following hypo-osmotic swelling and reduced the change in cell volume and the time to peak [Ca2+]i in response to hyper-osmotic stress, but did not affect the peak magnitudes of [Ca2+]i in those cells that did response. Co-treatment with IL-1Ra, thapsigargin, or Toxin B restored these responses to control levels. The effects were associated with alterations in F-actin organization. CONCLUSIONS IL-1 alters the normal volumetric and Ca2+ signaling response of chondrocytes to osmotic stress through mechanisms involving F-actin remodeling via small Rho GTPases. These findings provide further insights into the mechanisms by which IL-1 may interfere with normal physiologic processes in the chondrocyte, such as the adaptation or regulatory responses to mechanical and osmotic loading. PMID:18495501

  4. Aquaporin 1 contributes to chondrocyte apoptosis in a rat model of osteoarthritis.

    Science.gov (United States)

    Gao, Hangfei; Gui, Jiancao; Wang, Liming; Xu, Yan; Jiang, Yiqiu; Xiong, Mingyue; Cui, Yongguang

    2016-12-01

    Aquaporins (AQPs) have been found to be associated with a number of diseases. However, the role of AQP‑1 in the pathogenesis of osteoarthritis remains unclear. We previously found that AQP‑1 expression was upregulated in osteoarthritic cartilage and strongly correlated with caspase‑3 expression and activity. The aim of this study was to further investigate the association of AQP‑1 expression with chondrocyte apoptosis in a rat model of osteoarthritis, using RNA interference to knock down AQP‑1. For this purspose, 72 male Sprague‑Dawley rats were randomly assigned to 3 groups as follows: the control group not treated surgically (n=24), the sham‑operated group (n=24), and the osteoarthritis group (n=24). Osteoarthritis was induced by amputating the anterior cruciate ligament and medial collateral ligament and partially excising the medial meniscus. Chondrocytes from the rats with osteoarthritis were isolated and cultured. shRNAs were used to knock down AQP‑1 expression in the cultured chondrocytes. The expression of AQP‑1 and caspase‑3 was determined by reverse transcription-quantitative polymerase chain reaction. Caspase‑3 activity was measured using a caspase‑3 colorimetric assay. The rats in our model of osteoarthritis exhibited severe cartilage damage. The knockdown of AQP‑1 decreased caspase‑3 expression and activity in the cultured chondrocytes. In addition, the expression of AQP‑1 positively correlated with caspase‑3 expression and activity. Thus, the findings of our study, suggest that AQP‑1 promotes caspase‑3 activation and thereby contributes to chondrocyte apoptosis and to the development of osteoarthritis.

  5. Nanosized fibers' effect on adult human articular chondrocytes behavior

    Energy Technology Data Exchange (ETDEWEB)

    Stenhamre, Hanna [Biopolymer Technology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Gothenburg (Sweden); Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Gothenburg (Sweden); Thorvaldsson, Anna, E-mail: anna.thorvaldsson@swerea.se [Biopolymer Technology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Gothenburg (Sweden); Swerea IVF, Mölndal (Sweden); Enochson, Lars [Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Gothenburg (Sweden); Walkenström, Pernilla [Swerea IVF, Mölndal (Sweden); Lindahl, Anders [Department of Clinical Chemistry and Transfusion Medicine, Sahlgrenska University Hospital, Gothenburg (Sweden); Brittberg, Mats [Cartilage Research Unit, University of Gothenburg, Department Orthopaedics, Kungsbacka Hospital, Kungsbacka (Sweden); Gatenholm, Paul [Biopolymer Technology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Gothenburg (Sweden)

    2013-04-01

    Tissue engineering with chondrogenic cell based therapies is an expanding field with the intention of treating cartilage defects. It has been suggested that scaffolds used in cartilage tissue engineering influence cellular behavior and thus the long-term clinical outcome. The objective of this study was to assess whether chondrocyte attachment, proliferation and post-expansion re-differentiation could be influenced by the size of the fibers presented to the cells in a scaffold. Polylactic acid (PLA) scaffolds with different fiber morphologies were produced, i.e. microfiber (MS) scaffolds as well as nanofiber-coated microfiber scaffold (NMS). Adult human articular chondrocytes were cultured in the scaffolds in vitro up to 28 days, and the resulting constructs were assessed histologically, immunohistochemically, and biochemically. Attachment of cells and serum proteins to the scaffolds was affected by the architecture. The results point toward nano-patterning onto the microfibers influencing proliferation of the chondrocytes, and the overall 3D environment having a greater influence on the re-differentiation. In the efforts of finding the optimal scaffold for cartilage tissue engineering, studies as the current contribute to the knowledge of how to affect and control chondrocytes behavior. - Highlights: ► Chondrocyte behavior in nanofiber-coated microfiber versus microfiber scaffolds ► High porosity (> 90%) and large pore sizes (a few hundred μm) of nanofibrous scaffolds ► Proliferation enhanced by presence of nanofibers ► Differentiation not significantly affected ► Cell attachment improved in presence of both nanofibers and serum.

  6. Low oxygen reduces the modulation to an oxidative phenotype in monolayer-expanded chondrocytes.

    Science.gov (United States)

    Heywood, Hannah K; Lee, David A

    2010-01-01

    Autologous chondrocyte implantation requires a phase of in vitro cell expansion, achieved by monolayer culture under atmospheric oxygen levels. Chondrocytes reside under low oxygen conditions in situ and exhibit a glycolytic metabolism. However, oxidative phosphorylation rises progressively during culture, with concomitant reactive oxygen species production. We determine if the high oxygen environment in vitro provides the transformation stimulus. Articular chondrocytes were cultured in monolayer for up to 14 days under 2%, 5%, or 20% oxygen. Expansion under 2% and 5% oxygen reduced the rate at which the cells developed an oxidative phenotype compared to 20% oxygen. However, at 40 +/- 4 fmol cell(-1) h(-1) the oxygen consumption by chondrocytes expanded under 2% oxygen for 14 days was still 14 times the value observed for freshly isolated cells. Seventy-five to 78% of the increased oxygen consumption was accounted for by oxidative phosphorylation (oligomycin sensitive). Expansion under low oxygen also reduced cellular proliferation and 8-hydroxyguanosine release, a marker of oxidative DNA damage. However, these parameters remained elevated compared to freshly isolated cells. Thus, expansion under physiological oxygen levels reduces, but does not abolish, the induction of an oxidative energy metabolism. We conclude that simply transferring chondrocytes to low oxygen is not sufficient to either maintain or re-establish a normal energy metabolism. Furthermore, a hydrophobic polystyrene culture surface which promotes rounded cell morphology had no effect on the development of an oxidative metabolism. Although the shift towards an oxidative energy metabolism is often accompanied by morphological changes, this study does not support the hypothesis that it is driven by them.

  7. Sequential Vein Bypass Grafting is Not Associated with an Increase of Either In-hospital or Mid-term Adverse Events in Off-pump Coronary Artery Bypass Grafting

    Institute of Scientific and Technical Information of China (English)

    Fucheng Xiao; Jian Wang; Hengchao Wu; Hansong Sun

    2015-01-01

    Background:The impact of sequential vein bypass grafting on clinical outcomes is less known in off-pump coronary artery bypass grafting (CABG).We aimed to evaluate the effects of sequential vein bypass grafting on clinical outcomes in off-pump CABG.Methods:From October 2009 to September 2013 at the Fuwai Hospital,127 patients with at least one sequential venous graft were matched with 127 patients of individual venous grafts only,using propensity score matching method to obtain risk-adjusted outcome comparison.In-hospital measurement was composite outcome of in-hospital death,myocardial infarction (MI),stroke,requirement for intra-aortic ballon pump (IABP) assistance and prolonged ventilation.Major adverse cardiac events (MACEs:Death,MI or repeat revascularization) and angina recurrence were considered as mid-term endpoints.Results:No significant difference was observed among the groups in baseline characteristics.Intraoperative mean blood flow per vein graft was 40.4 ml in individual venous grafts groups versus 59.5 ml in sequential venous grafts groups (P < 0.001).There were no differences between individual and sequential venous grafts groups with regard to composite outcome of in-hospital mortality,MI,stroke,IABP assistance and prolonged ventilation (11.0% vs.14.2%,P =0.45).Individual in-hospital measurement also did not differ significantly between the two groups.At about four years follow-up,the survival estimates free from MACEs (92.5% vs.97.3%,P =0.36) and survival rates free of angina recurrence (80.9% vs.85.5%,P =0.48) were similar among individual and sequential venous grafts groups with a mean follow-up of 22.5 months.In the Cox regression analysis,sequential vein bypass grafting was not identified as an independent predictor of both MACEs and angina recurrence.Conclusions:Compared to individual vein bypass grafting,sequential vein bypass grafting was not associated with an increase of either in-hospital or mid-term adverse events in

  8. Photovoltaic private project at the Energy research Centre of the Netherlands (ECN). Four years of experience; PV-prive project bij ECN. Vier jaar ervaring

    Energy Technology Data Exchange (ETDEWEB)

    Jablonska, B.; Van Dijk, A.L.; Kaan, H.F.; Van Leeuwen, M.; De Boer, G.T.M.

    2004-10-01

    will continue at least for a period of five years. For this purpose, ECN has developed its own monitoring software. This software has also been used within other projects. The report treats the issue of compensation for the electricity supplied to the grid and the issue of electricity meters with registers turning backwards. Within the period of four years, a lot valuable knowledge has been gathered. This has been reached through continuous monitoring of practical experience since the beginning of the project as well as gaining information on non-technical aspects and operation of PV-systems by means of a questionnaire. The helpdesk and the service that shall stay available for five years have certainly contributed to this. Looking at all this experience it is clear that the project may be evaluated as successful. [Dutch] Het ECN PV-Prive project is opgezet om de toepassing van kleine netgekoppelde PV-systemen op bestaande woningen te onderzoeken. De ECN en NRG medewerkers hebben een mogelijkheid gekregen om een SunPower PV-systeem aan te schaffen in het kader van een aantrekkelijke financiele regeling. In ruil voor dit aanbod zouden zij aan het monitoringsproject meewerken en de opbrengsten van hun PV-systemen regelmatig aan ECN doorgeven. De rol van ECN in dit project is het monitoren en verspreiden van de ervaring opgedaan in de periode van de voorbereiding, installatie, nazorg en het gebruik van PV-systemen, alsook het verlenen van service en verstrekken van de informatie aan de deelnemers. Het monitoren van de opbrengsten en de kwaliteit van de PV-systemen is een uitgangspunt geweest voor vervolgonderzoeksprojecten. Binnen het PV-Prive project hebben 48 medewerkers van ECN en NRG een PV-systeem laten installeren. Na ruim drie jaar, tot en met juni 2004, is deze groep tot 36 deelnemers geslonken omdat een aantal deelnemers uit dienst is of het huis met het PV-systeem verkocht heeft. Of er een vergunning voor het plaatsen van het PV-systeem verleend werd, hing sterk

  9. Incorporation of hyaluronic acid into collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Tang Shunqing [Department of Biomedical Engineering, Jinan University, Guangzhou 510632 (China); Spector, Myron [Tissue Engineering, VA Boston Healthcare System, Boston, MA 02130 (United States)

    2007-09-15

    Hyaluronic acid (HA), a principal matrix molecule in many tissues, is present in high amounts in articular cartilage. HA contributes in unique ways to the physical behavior of the tissue, and has been shown to have beneficial effects on chondrocyte activity. The goal of this study was to incorporate graduated amounts of HA into type I collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis in vitro. The results demonstrated that the amount of contraction of HA/collagen scaffolds by adult canine articular chondrocytes increased with the HA content of the scaffolds. The greatest amount of chondrogenesis after two weeks was found in the scaffolds which had undergone the most contraction. HA can play a useful role in adjusting the mechanical behavior of tissue engineering scaffolds and chondrogenesis in chondrocyte-seeded scaffolds.

  10. Long-term and real-time monitoring of chondrocyte behavior synthesizing extracellular matrix with biologically coupled field effect transistor

    Science.gov (United States)

    Satake, Hiroto; Saito, Akiko; Mizuno, Shuichi; Kajisa, Taira; Sakata, Toshiya

    2017-04-01

    In this study, we report the differential measurement method of accurately monitoring cellular metabolism with a semiconductor-based field effect transistor (FET), focusing on the proliferation potency of chondrocytes utilized in the field of orthopedics. By adding growth factors to chondrocytes on the gate, cellular activity was induced and continuously monitored as a change in pH during a cellular respiration for ten days using the FET biosensor. Moreover, the electrical signal of the FET device reflected the reproduction property of chondrocytes to synthesize extracellular matrix (ECM). A platform based on the FET device is suitable as a noninvasive, real-time and long-term monitoring system for cellular functions; it will contribute to the elucidation of the mechanism of ECM synthesis by chondrocytes.

  11. SOX9 governs differentiation stage-specific gene expression in growth plate chondrocytes via direct concomitant transactivation and repression.

    Directory of Open Access Journals (Sweden)

    Victor Y L Leung

    2011-11-01

    Full Text Available Cartilage and endochondral bone development require SOX9 activity to regulate chondrogenesis, chondrocyte proliferation, and transition to a non-mitotic hypertrophic state. The restricted and reciprocal expression of the collagen X gene, Col10a1, in hypertrophic chondrocytes and Sox9 in immature chondrocytes epitomise the precise spatiotemporal control of gene expression as chondrocytes progress through phases of differentiation, but how this is achieved is not clear. Here, we have identified a regulatory element upstream of Col10a1 that enhances its expression in hypertrophic chondrocytes in vivo. In immature chondrocytes, where Col10a1 is not expressed, SOX9 interacts with a conserved sequence within this element that is analogous to that within the intronic enhancer of the collagen II gene Col2a1, the known transactivation target of SOX9. By analysing a series of Col10a1 reporter genes in transgenic mice, we show that the SOX9 binding consensus in this element is required to repress expression of the transgene in non-hypertrophic chondrocytes. Forced ectopic Sox9 expression in hypertrophic chondrocytes in vitro and in mice resulted in down-regulation of Col10a1. Mutation of a binding consensus motif for GLI transcription factors, which are the effectors of Indian hedgehog signaling, close to the SOX9 site in the Col10a1 regulatory element, also derepressed transgene expression in non-hypertrophic chondrocytes. GLI2 and GLI3 bound to the Col10a1 regulatory element but not to the enhancer of Col2a1. In addition to Col10a1, paired SOX9-GLI binding motifs are present in the conserved non-coding regions of several genes that are preferentially expressed in hypertrophic chondrocytes and the occurrence of pairing is unlikely to be by chance. We propose a regulatory paradigm whereby direct concomitant positive and negative transcriptional control by SOX9 ensures differentiation phase-specific gene expression in chondrocytes. Discrimination between

  12. Effects of RNAi-mediated inhibition of aggrecanase-1 and aggrecanase-2 on rat costochondral chondrocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    Zheng-hui WANG; Zhuang-qun YANG; Xi-jing HE; Li WANG; Li-xia LI; Jun-bo TU

    2008-01-01

    Aim:Failure of transplanted cartilage or allogenic chondrocytes is attributed mainly to immunological rejection and cartilage degradation.A major feature is the loss of aggrecan from the cartilage matrix,primarily due to the action of the specific proteinases aggrecanase-1 and aggrecanase-2.The aim of this in vitro study was to determine whether the specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi would mitigate aggrecan loss from cultured chondrocytes.Methods:Expression plasmid vectors of shRNA targeting aggrecanase-1 and aggrecanase-2 were constructed and transfected into cultured rattus costochondral chondrocytes.The transfected cells were induced with interleukin-1 β (IL-1β).Gene mRNA levels were analyzed by RT-PCR.Aggrecan and collagen Ⅱ content were measured by immunohistochemistry and Western blotting.Results:As the chondrocytes underwent dedifferentiation,agggrecanase-1 increased significantly.The specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi had no negative effect on the morphology and growth velocity of the chondrocytes.The mRNA of aggrecanase-1 and aggrecanase-2 decreased significantly.The α-2-macroglobulin expression level was increased by the shRNA specific for aggrecanase-1.Other genes of the chondrocytic extracellular matrix were not affected.RNAi significantly increased the aggrecan and collagen Ⅱ content of chondrocytes treated with IL-1β.Conclusion:The results suggest that inhibition of aggrecanase-1 and aggrecanase-2 by RNAi can mitigate aggrecan degradation,without interfering with chondrocytic gene phenotype recovery.RNAi technology can be a useful tool for studying degenerative processes in cartilage.

  13. Ear-Shaped Stable Auricular Cartilage Engineered from Extensively Expanded Chondrocytes in an Immunocompetent Experimental Animal Model.

    Science.gov (United States)

    Pomerantseva, Irina; Bichara, David A; Tseng, Alan; Cronce, Michael J; Cervantes, Thomas M; Kimura, Anya M; Neville, Craig M; Roscioli, Nick; Vacanti, Joseph P; Randolph, Mark A; Sundback, Cathryn A

    2016-02-01

    Advancement of engineered ear in clinical practice is limited by several challenges. The complex, largely unsupported, three-dimensional auricular neocartilage structure is difficult to maintain. Neocartilage formation is challenging in an immunocompetent host due to active inflammatory and immunological responses. The large number of autologous chondrogenic cells required for engineering an adult human-sized ear presents an additional challenge because primary chondrocytes rapidly dedifferentiate during in vitro culture. The objective of this study was to engineer a stable, human ear-shaped cartilage in an immunocompetent animal model using expanded chondrocytes. The impact of basic fibroblast growth factor (bFGF) supplementation on achieving clinically relevant expansion of primary sheep chondrocytes by in vitro culture was determined. Chondrocytes expanded in standard medium were either combined with cryopreserved, primary passage 0 chondrocytes at the time of scaffold seeding or used alone as control. Disk and human ear-shaped scaffolds were made from porous collagen; ear scaffolds had an embedded, supporting titanium wire framework. Autologous chondrocyte-seeded scaffolds were implanted subcutaneously in sheep after 2 weeks of in vitro incubation. The quality of the resulting neocartilage and its stability and retention of the original ear size and shape were evaluated at 6, 12, and 20 weeks postimplantation. Neocartilage produced from chondrocytes that were expanded in the presence of bFGF was superior, and its quality improved with increased implantation time. In addition to characteristic morphological cartilage features, its glycosaminoglycan content was high and marked elastin fiber formation was present. The overall shape of engineered ears was preserved at 20 weeks postimplantation, and the dimensional changes did not exceed 10%. The wire frame within the engineered ear was able to withstand mechanical forces during wound healing and neocartilage

  14. Anti-inflammatory activity of an ethanolic Caesalpinia sappan extract in human chondrocytes and macrophages

    Science.gov (United States)

    Wu, Shengqian Q; Otero, Miguel; Unger, Frank M; Goldring, Mary B; Phrutivorapongkul, Ampai; Chiari, Catharina; Kolb, Alexander; Viernstein, Helmut; Toegel, Stefan

    2012-01-01

    Aim of the study Caesalpinia sappan is a common remedy in Traditional Chinese Medicine and possesses diverse biological activities including anti-inflammatory properties. Osteoarthritis (OA) is a degenerative joint disease with an inflammatory component that drives the degradation of cartilage extracellular matrix. In order to provide a scientific basis for the applicability of Caesalpinia sappan in arthritic diseases, the present study aimed to assess the effects of an ethanolic Caesalpinia sappan extract (CSE) on human chondrocytes and macrophages. Materials and Methods Primary human chondrocytes were isolated from cartilage specimens of OA patients. Primary cells, SW1353 chondrocytes and THP-1 macrophages were serum-starved and pretreated with different concentrations of CSE prior to stimulation with 10 ng/ml of interleukin-1beta (IL-1ß) or lipopolysaccharide (LPS). Following viability tests, nitric oxide (NO) and tumor necrosis factor-alpha (TNF-α) were evaluated by Griess assay and ELISA, respectively. Using validated real-time PCR assays, mRNA levels of IL-1ß, TNF-α, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were quantified. SW1353 cells were cotransfected with a COX-2 luciferase reporter plasmid and nuclear factor-kappa-B (NF-κB) p50 and p65 expression vectors in the presence or absence of CSE. Results CSE dose-dependently inhibited the expression of pro-inflammatory cytokines IL-1ß and TNF-α in IL-1ß-stimulated chondrocytes and LPS-stimulated THP-1 macrophages. CSE further suppressed the synthesis of NO in primary OA chondrocytes by blocking iNOS mRNA expression. The inhibition of COX-2 transcription was found to be related with the CSE inhibition of the p65/p50-driven transactivation of the COX-2 promoter. Conclusions The present report is first to demonstrate the anti-inflammatory activity of CSE in an in vitro cell model of joint inflammation. CSE can effectively abrogate the IL-1ß-induced over-expression of

  15. Voltage-gated K+ currents in mouse articular chondrocytes regulate membrane potential.

    Science.gov (United States)

    Clark, Robert B; Hatano, Noriyuki; Kondo, Colleen; Belke, Darrell D; Brown, Barry S; Kumar, Sanjay; Votta, Bartholomew J; Giles, Wayne R

    2010-01-01

    Membrane currents and resting potential of isolated primary mouse articular chondrocytes maintained in monolayer cell culture for 1-9 days were recorded using patch clamp methods. Quantitative RT-PCR showed that the most abundantly expressed transcript of voltage-gated K(+) channels was for K(V)1.6, and immunological methods confirmed the expression of K(V)1.6 α-subunit proteins. These chondrocytes expressed a large time- and potential-dependent, Ca(2+)-independent 'delayed rectifier' K(+) current. Steady-state activation was well-fit by a Boltzmann function with a threshold near -50 mV, and a half-activation potential of -34.5 mV. The current was 50% blocked by 1.48 mM tetraethylammonium, 0.66 mM 4-aminopyridine and 20.6 nM α-dendrotoxin. The current inactivated very slowly at membrane potentials in the range of the resting potential of the chondrocytes. Resting membrane potential of the chondrocytes at room temperature (19-21°C) and in 5 mM external K(+) was -46.4 ± 1.3 mV (mean ± s.e.m; n = 23), near the 'foot' of the activation curve of this K(+) current. Resting potential was depolarized by an average of 4.2 ± 0.8 mV by 25 mM TEA, which blocked about 95% of the K(+) current. At a membrane potential of -50 mV, the apparent time constant of inactivation (tau(in)) was 37.9 s, and the 'steady-state' current level was 19% of that at a holding potential of -90 mV; at -40 mV, tau(in) was 20.3 s, and 'steady-state' current was 5% of that at -90 mV. These results demonstrate that in these primary cultured, mouse articular chondrocytes steady-state activation of a voltage-gated K(+) current contributes to resting membrane potential. However, this current is also likely to have a significant physiological role in repolarizing the chondrocyte following depolarizing stimuli that might occur in conditions of membrane stretch. For example, activation of TRP('transient receptor potential') non-specific cation channels in these cells during cyclic loading and unloading

  16. Osthole Inhibits Proliferation and Induces Catabolism in Rat Chondrocytes and Cartilage Tissue

    Directory of Open Access Journals (Sweden)

    Guoqing Du

    2015-08-01

    Full Text Available Background/Aims: Cartilage destruction is thought to be the major mediator of osteoarthritis. Recent studies suggest that inhibition of subchrondral bone loss by anti-osteoporosis (OP drug can protect cartilige erosion. Osthole, as a promising agent for treating osteoporosis, may show potential in treating osteoarthritis. The purpose of this study was to investigate whether Osthole affects the proliferation and catabolism of rat chondrocytes, and the degeneration of cartilage explants. Methods: Rat chondrocytes were treated with Osthole (0 μM, 6.25 μM, 12.5 μM, and 25 μM with or without IL1-β (10ng/ml for 24 hours. The expression levels of type II collagen and MMP13 were detected by western Blot. Marker genes for chondrocytes (A-can and Sox9, matrix metalloproteinases (MMPs, aggrecanases (ADAMTS5 and genes implicated in extracellular matrix catabolism were evaluated by qPCR. Cell proliferation was assessed by measuring proliferating cell nuclear antigen (PCNA expression and fluorescence activated cell sorter. Wnt7b/β-catenin signaling was also investigated. Cartilage explants from two-week old SD rats were cultured with IL-1β, Osthole and Osthole plus IL-1β for four days and glycosaminoglycan (GAG synthesis was assessed with toluidine blue staining and Safranine O/Fast Green FCF staining, collagen type II expression was detected by immunofuorescence. Results: Osthole reduced expression of chondrocyte markers and increased expression of MMP13, ADAMTS5 and MMP9 in a dose-dependent manner. Catabolic gene expression levels were further improved by Osthole plus IL-1β. Osthole inhibited chondrocyte proliferation. GAG synthesis and type II collagen were decreased in both the IL-1β groups and the Osthole groups, and significantly reduced by Osthole plus IL-1β. Conclusions: Our data suggested that Osthole increases the catabolism of rat chondrocytes and cartilage explants, this effect might be mediated through inhibiting Wnt7b

  17. High glucose mediates endothelial-to-chondrocyte transition in human aortic endothelial cells

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    Tang Rining

    2012-09-01

    Full Text Available Abstract Background Vascular calcification is one of the common complications in diabetes mellitus. Many studies have shown that high glucose (HG caused cardiovascular calcification, but its underlying mechanism is not fully understood. Recently, medial calcification has been most commonly described in the vessels of patients with diabetes. Chondrocytes were involved in the medial calcification. Recent studies have shown that the conversion into mesenchymal stem cells (MSCs via the endothelial-to-mesenchymal transition (EndMT could be triggered in chondrocytes. Our previous research has indicated that HG induced EndMT in human aortic endothelial cells (HAECs. Therefore, we addressed the question of whether HG-induced EndMT could be transitioned into MSCs and differentiated into chondrocytes. Methods HAECs were divided into three groups: a normal glucose (NG group, HG group (30 mmol/L, and mannitol (5.5 mmol/L NG + 24.5 mmol/L group. Pathological changes were investigated using fluorescence microscopy and electron microscopy. Immunofluorescence staining was performed to detect the co-expression of endothelial markers, such as CD31, and fibroblast markers, such as fibroblast-specific protein 1 (FSP-1. The expression of FSP-1 was detected by real time-PCR and western blots. Endothelial-derived MSCs were grown in MSC medium for one week. The expression of the MSCs markers STRO-1, CD44, CD10 and the chondrocyte marker SOX9 was detected by immunofluorescence staining and western blots. Chondrocyte expression was detected by alcian blue staining. Calcium deposits were analyzed by alizarin red staining. Results The incubation of HAECs exposed to HG resulted in a fibroblast-like phenotype. Double staining of the HAECs indicated a co-localization of CD31 and FSP-1. The expression of FSP-1 was significantly increased in the HG group, and the cells undergoing EndMT also expressed STRO-1, CD44 and SOX9 compared with the controls (P  Conclusions Our

  18. [The synergistic effect of amygdalin and HSYA on the IL-1beta induced endplate chondrocytes of rat intervertebral discs].

    Science.gov (United States)

    Niu, Kai; Zhao, Yong-Jian; Zhang, Lei; Li, Chen-Guang; Wang, Yong-Jun; Zheng, Wei-Chao

    2014-08-01

    The effect of amygdalin joint hydroxysafflor yellow A (HSYA) on the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and the possible mechanism were studied and explored. Chondrocytes were obtained from endplate of one-month SD rat intervertebral discs and cultured primary endplate chondrocytes. After identification, they were divided into normal group, induced group, amygdalin group, HSYA group and combined group. CCK-8 kit was adopted to detect the proliferation of the endplate chondrocytes. FCM was measured to detect the apoptosis. Real-time PCR method was adopted to observe the mRNA expression of Aggrecan, Col 2 alpha1, Col 10 alpha1, MMP-13 and the inflammatory cytokines IL-1beta. The protein expression of Col II, Col X was tested through immunofluorescence. Compared with the normal group, the proliferation of the endplate chondrocytes decreased while the apoptosis increased (P amygdalin group, HSYA group, the combined group could inhibit the apoptosis and promote the proliferation (P amygdalin and HSYA. Amygdalin joint HSYA could inhibit the degeneration of the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and better than the single use of amygdalin or HSYA.

  19. Activation of α2A-adrenergic signal transduction in chondrocytes promotes degenerative remodelling of temporomandibular joint

    Science.gov (United States)

    Jiao, Kai; Zeng, Guang; Niu, Li-Na; Yang, Hong-xu; Ren, Gao-tong; Xu, Xin-yue; Li, Fei-fei; Tay, Franklin R.; Wang, Mei-qing

    2016-01-01

    This study tested whether activation of adrenoreceptors in chondrocytes has roles in degenerative remodelling of temporomandibular joint (TMJ) and to determine associated mechanisms. Unilateral anterior crossbite (UAC) was established to induce TMJ degeneration in rats. Saline vehicle, α2- and β-adrenoreceptor antagonists or agonists were injected locally into the TMJ area of UAC rats. Cartilage degeneration, subchondral bone microarchitecture and the expression of adrenoreceptors, aggrecans, matrix metalloproteinases (MMPs) and RANKL by chondrocytes were evaluated. Chondrocytes were stimulated by norepinephrine to investigate signal transduction of adrenoreceptors. Increased α2A-adrenoreceptor expression was observed in condylar cartilage of UAC rats, together with cartilage degeneration and subchondral bone loss. Norepinephrine depresses aggrecans expression but stimulates MMP-3, MMP-13 and RANKL production by chondrocytes through ERK1/2 and PKA pathway; these effects were abolished by an α2A-adrenoreceptor antagonist. Furthermore, inhibition of α2A-adrenoreceptor attenuated degenerative remodelling in the condylar cartilage and subchondral bone, as revealed by increased cartilage thickness, proteoglycans and aggrecan expression, and decreased MMP-3, MMP-13 and RANKL expressions in cartilage, increased BMD, BV/TV, and decreased Tb.Sp in subchondral bone. Conversely, activation of α2A-adrenoreceptor intensified aforementioned degenerative changes in UAC rats. It is concluded that activation of α2A-adrenergic signal in chondrocytes promotes TMJ degenerative remodelling by chondrocyte-mediated pro-catabolic activities. PMID:27452863

  20. The Effect of Chondroitin Sulphate and Hyaluronic Acid on Chondrocytes Cultured within a Fibrin-Alginate Hydrogel

    Directory of Open Access Journals (Sweden)

    Christopher J. Little

    2014-09-01

    Full Text Available Osteoarthritis is a painful degenerative joint disease that could be better managed if tissue engineers can develop methods to create long-term engineered articular cartilage tissue substitutes. Many of the tissue engineered cartilage constructs currently available lack the chemical stimuli and cell-friendly environment that promote the matrix accumulation and cell proliferation needed for use in joint cartilage repair. The goal of this research was to test the efficacy of using a fibrin-alginate hydrogel containing hyaluronic acid (HA and/or chondroitin sulphate (CS supplements for chondrocyte culture. Neonatal porcine chondrocytes cultured in fibrin-alginate hydrogels retained their phenotype better than chondrocytes cultured in monolayer, as evidenced by analysis of their relative expression of type II versus type I collagen mRNA transcripts. HA or CS supplementation of the hydrogels increased matrix glycosaminoglycan (GAG production during the first week of culture. However, the effects of these supplements on matrix accumulation were not additive and were no longer observed after two weeks of culture. Supplementation of the hydrogels with CS or a combination of both CS and HA increased the chondrocyte cell population after two weeks of culture. Statistical analysis indicated that the HA and CS treatment effects on chondrocyte numbers may be additive. This research suggests that supplementation with CS and/or HA has positive effects on cartilage matrix production and chondrocyte proliferation in three-dimensional (3D fibrin-alginate hydrogels.

  1. Prolyl Hydroxylase Domain-Containing Protein 2 (Phd2) Regulates Chondrocyte Differentiation and Secondary Ossification in Mice

    Science.gov (United States)

    Cheng, Shaohong; Aghajanian, Patrick; Pourteymoor, Sheila; Alarcon, Catrina; Mohan, Subburaman

    2016-01-01

    Endochondral ossification plays an important role in the formation of the primary ossification centers (POCs) and secondary ossification centers (SOCs) of mammalian long bones. However, the molecular mechanisms that regulate POC and SOC formation are different. We recently demonstrated that Prolyl Hydroxylase Domain-containing Protein 2 (Phd2) is a key mediator of vitamin C effects on bone. We investigated the role of Phd2 on endochondral ossification of the epiphyses by conditionally deleting the Phd2 gene in osteoblasts and chondrocytes. We found that the deletion of Phd2 in osteoblasts did not cause changes in bone parameters in the proximal tibial epiphyses in 5 week old mice. In contrast, deletion of Phd2 in chondrocytes resulted in increased bone mass and bone formation rate (normalized to tissue volume) in long bone epiphyses, indicating that Phd2 expressed in chondrocytes, but not osteoblasts, negatively regulates secondary ossification of epiphyses. Phd2 deletion in chondrocytes elevated mRNA expression of hypoxia-inducible factor (HIF) signaling molecules including Hif-1α, Hif-2α, Vegfa, Vegfb, and Epo, as well as markers for chondrocyte hypertrophy and mineralization such as Col10, osterix, alkaline phosphatase, and bone sialoprotein. These data suggest that Phd2 expressed in chondrocytes inhibits endochondral ossification at the epiphysis by suppressing HIF signaling pathways. PMID:27775044

  2. Gene Modification of Mesenchymal Stem Cells and Articular Chondrocytes to Enhance Chondrogenesis

    Directory of Open Access Journals (Sweden)

    Saliya Gurusinghe

    2014-01-01

    Full Text Available Current cell based treatment for articular cartilage and osteochondral defects are hampered by issues such as cellular dedifferentiation and hypertrophy of the resident or transplanted cells. The reduced expression of chondrogenic signalling molecules and transcription factors is a major contributing factor to changes in cell phenotype. Gene modification of chondrocytes may be one approach to redirect cells to their primary phenotype and recent advances in nonviral and viral gene delivery technologies have enabled the expression of these lost factors at high efficiency and specificity to regain chondrocyte function. This review focuses on the various candidate genes that encode signalling molecules and transcription factors that are specific for the enhancement of the chondrogenic phenotype and also how epigenetic regulators of chondrogenesis in the form of microRNA may also play an important role.

  3. Stem cells catalyze cartilage formation by neonatal articular chondrocytes in 3D biomimetic hydrogels

    Science.gov (United States)

    Lai, Janice H.; Kajiyama, Glen; Smith, Robert Lane; Maloney, William; Yang, Fan

    2013-12-01

    Cartilage loss is a leading cause of disability among adults and effective therapy remains elusive. Neonatal chondrocytes (NChons) are an attractive allogeneic cell source for cartilage repair, but their clinical translation has been hindered by scarce donor availability. Here we examine the potential for catalyzing cartilage tissue formation using a minimal number of NChons by co-culturing them with adipose-derived stem cells (ADSCs) in 3D hydrogels. Using three different co-culture models, we demonstrated that the effects of co-culture on cartilage tissue formation are dependent on the intercellular distance and cell distribution in 3D. Unexpectedly, increasing ADSC ratio in mixed co-culture led to increased synergy between NChons and ADSCs, and resulted in the formation of large neocartilage nodules. This work raises the potential of utilizing stem cells to catalyze tissue formation by neonatal chondrocytes via paracrine signaling, and highlights the importance of controlling cell distribution in 3D matrices to achieve optimal synergy.

  4. Flavonoid Compound Icariin Activates Hypoxia Inducible Factor-1α in Chondrocytes and Promotes Articular Cartilage Repair.

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    Pengzhen Wang

    Full Text Available Articular cartilage has poor capability for repair following trauma or degenerative pathology due to avascular property, low cell density and migratory ability. Discovery of novel therapeutic approaches for articular cartilage repair remains a significant clinical need. Hypoxia is a hallmark for cartilage development and pathology. Hypoxia inducible factor-1alpha (HIF-1α has been identified as a key mediator for chondrocytes to response to fluctuations of oxygen availability during cartilage development or repair. This suggests that HIF-1α may serve as a target for modulating chondrocyte functions. In this study, using phenotypic cellular screen assays, we identify that Icariin, an active flavonoid component from Herba Epimedii, activates HIF-1α expression in chondrocytes. We performed systemic in vitro and in vivo analysis to determine the roles of Icariin in regulation of chondrogenesis. Our results show that Icariin significantly increases hypoxia responsive element luciferase reporter activity, which is accompanied by increased accumulation and nuclear translocation of HIF-1α in murine chondrocytes. The phenotype is associated with inhibiting PHD activity through interaction between Icariin and iron ions. The upregulation of HIF-1α mRNA levels in chondrocytes persists during chondrogenic differentiation for 7 and 14 days. Icariin (10-6 M increases the proliferation of chondrocytes or chondroprogenitors examined by MTT, BrdU incorporation or colony formation assays. Icariin enhances chondrogenic marker expression in a micromass culture including Sox9, collagen type 2 (Col2α1 and aggrecan as determined by real-time PCR and promotes extracellular matrix (ECM synthesis indicated by Alcian blue staining. ELISA assays show dramatically increased production of aggrecan and hydroxyproline in Icariin-treated cultures at day 14 of chondrogenic differentiation as compared with the controls. Meanwhile, the expression of chondrocyte catabolic

  5. The ECM-Cell Interaction of Cartilage Extracellular Matrix on Chondrocytes

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    Yue Gao

    2014-01-01

    Full Text Available Cartilage extracellular matrix (ECM is composed primarily of the network type II collagen (COLII and an interlocking mesh of fibrous proteins and proteoglycans (PGs, hyaluronic acid (HA, and chondroitin sulfate (CS. Articular cartilage ECM plays a crucial role in regulating chondrocyte metabolism and functions, such as organized cytoskeleton through integrin-mediated signaling via cell-matrix interaction. Cell signaling through integrins regulates several chondrocyte functions, including differentiation, metabolism, matrix remodeling, responses to mechanical stimulation, and cell survival. The major signaling pathways that regulate chondrogenesis have been identified as wnt signal, nitric oxide (NO signal, protein kinase C (PKC, and retinoic acid (RA signal. Integrins are a large family of molecules that are central regulators in multicellular biology. They orchestrate cell-cell and cell-matrix adhesive interactions from embryonic development to mature tissue function. In this review, we emphasize the signaling molecule effect and the biomechanics effect of cartilage ECM on chondrogenesis.

  6. Green fluorescent protein as marker in chondrocytes overexpressing human insulin-like growth factor-1 for repair of articular cartilage defects in rabbits

    Institute of Scientific and Technical Information of China (English)

    ZHANG Shao-kun; LIU Yi; SONG Zhi-ming; FU Chang-feng; XU Xin-xiang

    2007-01-01

    Objective:To label the primary articular chondrocytes overexpressing human insulin-like growth factor ( hIGF-1 ) with green fluorescent protein (GFP) for repair of articular cartilage defects in rabbits. Methods:GFP cDNA was inserted into pcDNA3.1-hIGF-1 to label the expression vector.The recombinant vector,pcGI,a mammalian expression vector with multiple cloning sites under two respective cytomegalovirus promoters/enhancers,was transfected into the primary articular chondrocytes with the help of lipofectamine.After the positive cell clones were selected by G418,G418-resistant chondrocytes were cultured in medium for 4 weeks.The stable expression of hIGF-1 in the articular chondrocytes was determined by in situ hybridization and immunocytochemical analysis and the GFP was confirmed under a fluorescence microscope. Methyl thiazolyl tetrazolium (MTT) and flow cytometer methods were employed to determine the effect of transfection on proliferation of chondrocytes. Gray value was used to analyze quantitatively the expression of type Ⅱ collagen. Results:The expression of hIGF-1 and GFP was confirmed in transfected chondrocytes by in situ hybridization, immunocytochemical analysis and fluorescence microscope observation. Green articular chondrocytes overexpressing hIGF-1 could expand and maintain their chondrogenic phenotypes for more than 4 weeks.After the transfection of IGF-1,the proliferation of chondrocytes was enhanced and the chondrocytes could effectively maintain the expression of type Ⅱ collagen. Conclusions:The hIGF-1 eukaryotic expression vector containing GFP marker gene has been successfully constructed.GFP,which can be visualized in real time and in situ, is stably expressed in articular chondrocytes overexpressing hIGF-1.The labeled articular chondrocytes overexpressing hIGF-1 can be applied in cell-mediated gene therapy as well as for other biomedical purposes of transgenic chondrocytes.

  7. Effect of polystyrene and polyether imide cell culture inserts with different roughness on chondrocyte metabolic activity and gene expression profiles of aggrecan and collagen.

    Science.gov (United States)

    König, Josephine; Kohl, Benjamin; Kratz, Karl; Jung, Friedrich; Lendlein, Andreas; Ertel, Wolfgang; Schulze-Tanzil, Gundula

    2013-01-01

    In vitro cultured autologous chondrocytes can be used for implantation to support cartilage repair. For this purpose, a very small number of autologous cells harvested from a biopsy have to be expanded in monolayer culture. Commercially available polymer surfaces lead to chondrocyte dedifferentiation. Hence, the demanding need for optimized polymers and surface topologies supporting chondrocytes' differentiated phenotypes in vitro arises. In this study we explored the effect of tailored cell culture plate inserts prepared from polystyrene (PS) and polyether imide (PEI) exhibiting three different roughness levels (R0, RI, RII) on chondrocyte morphology, metabolism and gene expression profile. As a control, commercially available tissue culture plastic (TCP) dishes were included. Primary porcine articular chondrocytes were seeded on tailored PS and PEI inserts with three different roughness levels. The metabolic activity of the chondrocytes was determined after 24 hours using alamar blue assay. Chondrocyte gene expression profiles (aggrecan, type I and type II collagen) were monitored after 48 hours using Real Time Detection (RTD)-PCR. Chondrocytes cultured on PS and PEI surfaces formed cell clusters after 24 and 48 hours, which was not observed on TCP. The metabolic activity of chondrocytes cultured on PS was lower than of chondrocytes cultured on PEI, but also lower than on TCP. Gene expression analyses revealed an elevated expression of cartilage-specific aggrecan and an impaired expression of both collagen types by chondrocytes on PS and PEI compared with TCP. In summary, PEI is a biocompatible biomaterial suitable for chondrocyte culturing, which can be further chemically functionalized for generating specific surface interactions or covalent binding of biomolecules.

  8. Prevention of primary vascular graft infection with silver-coated polyester graft in a porcine model

    DEFF Research Database (Denmark)

    Gao, H; Sandermann, J; Prag, J;

    2010-01-01

    To evaluate the efficacy of a silver-coated vascular polyester graft in the prevention of graft infection after inoculation with Staphylococcus aureus in a porcine model.......To evaluate the efficacy of a silver-coated vascular polyester graft in the prevention of graft infection after inoculation with Staphylococcus aureus in a porcine model....

  9. Piperine inhibits IL-β induced expression of inflammatory mediators in human osteoarthritis chondrocyte.

    Science.gov (United States)

    Ying, Xiaozhou; Chen, Xiaowei; Cheng, Shaowen; Shen, Yue; Peng, Lei; Xu, Hua Zi

    2013-10-01

    Black pepper (Piper nigrum) is a common remedy in Traditional Chinese Medicine and possesses diverse biological activities including anti-inflammatory properties. Osteoarthritis (OA) is a degenerative joint disease with an inflammatory component that drives the degradation of cartilage extracellular matrix. The present study aimed to assess the effects of piperine, the active phenolic component in black pepper extract, on human OA chondrocytes. In this study, human OA chondrocytes were pretreated with piperine at 10, 50 or 100μg/ml and subsequently stimulated with IL-1β (5ng/ml) for 24h. Production of PGE2 and NO was evaluated by the Griess reaction and an ELISA. Gene expression of MMP-3, MMP-13, iNOS and COX-2 was measured by real-time PCR. MMP-3 and MMP-13 proteins in culture medium were determined using cytokine-specific ELISA. Western immunoblotting was used to analyze the iNOS and COX-2 protein production in the culture medium. The regulation of NF-kB activity and the degradation of IkB were explored using luciferase and Western immunoblotting, respectively. We found that piperine inhibited the production of PGE2 and NO induced by IL-1β. Piperine significantly decreased the IL-1β-stimulated gene expression and production of MMP-3, MMP-13, iNOS and COX-2 in human OA chondrocytes. Piperine inhibited the IL-1β-mediated activation of NF-κB by suppressing the degradation of its inhibitory protein IκBα in the cytoplasm. The present report is first to demonstrate the anti-inflammatory activity of piperine in human OA chondrocytes. Piperine can effectively abrogate the IL-1β-induced over-expression of inflammatory mediators; suggesting that piperine may be a potential agent in the treatment of OA.

  10. Dexamethasone stimulates expression of C-type Natriuretic Peptide in chondrocytes

    Directory of Open Access Journals (Sweden)

    Beier Frank

    2006-11-01

    Full Text Available Abstract Background Growth of endochondral bones is regulated through the activity of cartilaginous growth plates. Disruption of the physiological patterns of chondrocyte proliferation and differentiation – such as in endocrine disorders or in many different genetic diseases (e.g. chondrodysplasias – generally results in dwarfism and skeletal defects. For example, glucocorticoid administration in children inhibits endochondral bone growth, but the molecular targets of these hormones in chondrocytes remain largely unknown. In contrast, recent studies have shown that C-type Natriuretic Peptide (CNP is an important anabolic regulator of cartilage growth, and loss-of-function mutations in the human CNP receptor gene cause dwarfism. We asked whether glucocorticoids could exert their activities by interfering with the expression of CNP or its downstream signaling components. Methods Primary mouse chondrocytes in monolayer where incubated with the synthetic glucocorticoid Dexamethasone (DEX for 12 to 72 hours. Cell numbers were determined by counting, and real-time PCR was performed to examine regulation of genes in the CNP signaling pathway by DEX. Results We show that DEX does influence expression of key genes in the CNP pathway. Most importantly, DEX significantly increases RNA expression of the gene encoding CNP itself (Nppc. In addition, DEX stimulates expression of Prkg2 (encoding cGMP-dependent protein kinase II and Npr3 (natriuretic peptide decoy receptor genes. Conversely, DEX was found to down-regulate the expression of the gene encoding its receptor, Nr3c1 (glucocorticoid receptor, as well as the Npr2 gene (encoding the CNP receptor. Conclusion Our data suggest that the growth-suppressive activities of DEX are not due to blockade of CNP signaling. This study reveals a novel, unanticipated relationship between glucocorticoid and CNP signaling and provides the first evidence that CNP expression in chondrocytes is regulated by endocrine

  11. Substrate Stiffness Controls Osteoblastic and Chondrocytic Differentiation of Mesenchymal Stem Cells without Exogenous Stimuli

    Science.gov (United States)

    Hyzy, Sharon L.; Doroudi, Maryam; Williams, Joseph K.; Gall, Ken; Boyan, Barbara D.; Schwartz, Zvi

    2017-01-01

    Stem cell fate has been linked to the mechanical properties of their underlying substrate, affecting mechanoreceptors and ultimately leading to downstream biological response. Studies have used polymers to mimic the stiffness of extracellular matrix as well as of individual tissues and shown mesenchymal stem cells (MSCs) could be directed along specific lineages. In this study, we examined the role of stiffness in MSC differentiation to two closely related cell phenotypes: osteoblast and chondrocyte. We prepared four methyl acrylate/methyl methacrylate (MA/MMA) polymer surfaces with elastic moduli ranging from 0.1 MPa to 310 MPa by altering monomer concentration. MSCs were cultured in media without exogenous growth factors and their biological responses were compared to committed chondrocytes and osteoblasts. Both chondrogenic and osteogenic markers were elevated when MSCs were grown on substrates with stiffness <10 MPa. Like chondrocytes, MSCs on lower stiffness substrates showed elevated expression of ACAN, SOX9, and COL2 and proteoglycan content; COMP was elevated in MSCs but reduced in chondrocytes. Substrate stiffness altered levels of RUNX2 mRNA, alkaline phosphatase specific activity, osteocalcin, and osteoprotegerin in osteoblasts, decreasing levels on the least stiff substrate. Expression of integrin subunits α1, α2, α5, αv, β1, and β3 changed in a stiffness- and cell type-dependent manner. Silencing of integrin subunit beta 1 (ITGB1) in MSCs abolished both osteoblastic and chondrogenic differentiation in response to substrate stiffness. Our results suggest that substrate stiffness is an important mediator of osteoblastic and chondrogenic differentiation, and integrin β1 plays a pivotal role in this process. PMID:28095466

  12. A mutation in TRPV4 results in altered chondrocyte calcium signaling in severe metatropic dysplasia.

    Science.gov (United States)

    Hurd, Lauren; Kirwin, Susan M; Boggs, Mary; Mackenzie, William G; Bober, Michael B; Funanage, Vicky L; Duncan, Randall L

    2015-10-01

    Transient receptor potential cation channel, subfamily V, member 4 (TRPV4) is a polymodal modulated non-selective cation channel required for normal development and maintenance of bone and cartilage. Heterozygous mutations of this channel cause a variety of channelopathies, including metatropic dysplasia (MD). We analyzed the effect of a novel TRPV4 mutation c.2398G>A, p.Gly800Asp on intracellular calcium ([Ca(2+) ]i ) regulation in chondrocytes and compared this response to chondrocytes with a frequently observed mutation, c.2396C>T, p.Pro799Leu. We observed temperature-dependent [Ca(2+) ]i oscillations in both intact and MD chondrocytes however, MD mutations exhibited increased peak magnitudes of [Ca(2+) ]i during oscillations. We also found increased baseline [Ca(2+) ]i in MD primary cells, as well as increased [Ca(2+) ]i response to either hypotonic swelling or the TRVP4-specific agonist, GSK1016790A. Oscillations and stimulation responses were blocked with the TRPV4-specific antagonist, GSK205. Analysis of [Ca(2+) ]i response kinetics showed that MD chondrocytes had increased frequency of temperature-sensitive oscillations, and the magnitude and duration of [Ca(2+) ]i responses to given stimuli. Duration of the response of the p.Gly800Asp mutation to stimulation was greater than for the p.Pro799Leu mutation. These experiments show that this region of the channel is essential for proper [Ca(2+) ]i regulation. These studies of primary cells from patients show how both mutant and WT TRPV4 channels regulate cartilage and bone development. © 2015 Wiley Periodicals, Inc.

  13. Carnosol and Related Substances Modulate Chemokine and Cytokine Production in Macrophages and Chondrocytes

    Directory of Open Access Journals (Sweden)

    Joseph Schwager

    2016-04-01

    Full Text Available Phenolic diterpenes present in Rosmarinus officinalis and Salvia officinalis have anti-inflammatory and chemoprotective effects. We investigated the in vitro effects of carnosol (CL, carnosic acid (CA, carnosic acid-12-methylether (CAME, 20-deoxocarnosol and abieta-8,11,13-triene-11,12,20-triol (ABTT in murine macrophages (RAW264.7 cells and human chondrocytes. The substances concentration-dependently reduced nitric oxide (NO and prostaglandin E2 (PGE2 production in LPS-stimulated macrophages (i.e., acute inflammation. They significantly blunted gene expression levels of iNOS, cytokines/interleukins (IL-1α, IL-6 and chemokines including CCL5/RANTES, CXCL10/IP-10. The substances modulated the expression of catabolic and anabolic genes in chondrosarcoma cell line SW1353 and in primary human chondrocytes that were stimulated by IL-1β (i.e., chronic inflammation In SW1353, catabolic genes like MMP-13 and ADAMTS-4 that contribute to cartilage erosion were down-regulated, while expression of anabolic genes including Col2A1 and aggrecan were shifted towards pre-pathophysiological homeostasis. CL had the strongest overall effect on inflammatory mediators, as well as on macrophage and chondrocyte gene expression. Conversely, CAME mainly affected catabolic gene expression, whereas ABTT had a more selectively altered interleukin and chemokine gene exprssion. CL inhibited the IL-1β induced nuclear translocation of NF-κBp65, suggesting that it primarily regulated via the NF-κB signalling pathway. Collectively, CL had the strongest effects on inflammatory mediators and chondrocyte gene expression. The data show that the phenolic diterpenes altered activity pattern of genes that regulate acute and chronic inflammatory processes. Since the substances affected catabolic and anabolic gene expression in cartilage cells in vitro, they may beneficially act on the aetiology of osteoarthritis.

  14. Increased Production of Clusterin in Biopsies of Repair Tissue following Autologous Chondrocyte Implantation

    Science.gov (United States)

    Malda, Jos; Richardson, James B.; Roberts, Sally

    2013-01-01

    Objective. To characterize the immunolocalization of clusterin in the repair cartilage of patients having undergone autologous chondrocyte implantation (ACI) and evaluate correlation to clinical outcome. Design. Full-depth core biopsies of repair tissue were obtained from 38 patients who had undergone ACI at an average of 18 ± 13 months previously (range 8-67 months). The biopsies were snap frozen, cryosectioned, and clusterin production immunolocalized using a specific monoclonal clusterin antibody and compared with normal and osteoarthritic cartilage. Clinical outcome was assessed from patients preoperatively, at the time of biopsy, and annually postoperatively. Results. Intensity of immunostaining for clusterin decreased with age in healthy cartilage tissue. Clusterin was detected to a variable degree in 37 of the 38 ACI cartilage biopsies, in single and clustered chondrocytes, in the pericellular capsule and the cartilage extracellular matrix, as well as the osteocytes and osteoid within the bone. Chondrocytes in hyaline repair tissue were significantly more immunopositive than those in fibrocartilage repair tissue. Clinical outcome improved significantly post-ACI, but did not correlate with the presence of clusterin in the repair tissue. Conclusions. These results demonstrate the presence of clusterin in actively repairing human cartilage and indicate a different distribution of clusterin in this tissue compared to normal cartilage. Variability in clusterin staining in the repair tissue could indicate different states of chondrogenic differentiation. The clinical significance of clusterin within repair tissue is difficult to assess, although the ideal functioning repair tissue morphology should resemble that of healthy adult cartilage. PMID:26069669

  15. Investigation of the direct effects of salmon calcitonin on human osteoarthritic chondrocytes

    Directory of Open Access Journals (Sweden)

    Pedersen Christian

    2010-04-01

    Full Text Available Abstract Background Calcitonin has been demonstrated to have chondroprotective effects under pre-clinical settings. It is debated whether this effect is mediated through subchondral-bone, directly on cartilage or both in combination. We investigated possible direct effects of salmon calcitonin on proteoglycans and collagen-type-II synthesis in osteoarthritic (OA cartilage. Methods Human OA cartilage explants were cultured with salmon calcitonin [100 pM-100 nM]. Direct effects of calcitonin on articular cartilage were evaluated by 1 measurement of proteoglycan synthesis by incorporation of radioactive labeled 35SO4 [5 μCi] 2 quantification of collagen-type-II formation by pro-peptides of collagen type II (PIINP ELISA, 3 QPCR expression of the calcitonin receptor in OA chondrocytes using four individual primer pairs, 4 activation of the cAMP signaling pathway by EIA and, 5 investigations of metabolic activity by AlamarBlue. Results QPCR analysis and subsequent sequencing confirmed expression of the calcitonin receptor in human chondrocytes. All doses of salmon calcitonin significantly elevated cAMP levels (P 35SO4 incorporation, with a 96% maximal induction at 10 nM (P Conclusion Calcitonin treatment increased proteoglycan and collagen synthesis in human OA cartilage. In addition to its well-established effect on subchondral bone, calcitonin may prove beneficial to the management of joint diseases through direct effects on chondrocytes.

  16. Effects of cytokines, growth factors and drugs on matrix metalloproteinases activities of osteoarthritic chondrocytes and synoviocytes

    Institute of Scientific and Technical Information of China (English)

    GUAN Jian-long; HAN Xing-hai; SHI Gui-ying; YUAN Guo-hua

    2001-01-01

    Objective: To evaluate the effects of some cytokines, TGF-β1 and drugs on matrix metalloproteinases (MMPs) activities in culture medium of arthritic chondrocytes and synoviocytes. Methods: The chondrocyte and synoviocyte monolayers isolated from the cartilages and synovial fluids in 10 knee OA patients were treated with IL-1β TGF-β1, TNF-α, diclofenac acid, dexamethasone or doxycycline individually and together for 72 h. Zymography was used to determine the activities of MMP-2 and -9. Results: The chondrocyte monolayers produced MMP-2 and -9, while the synoviocytes only produced MMP-2. The MMP-9 activity was markedly enhanced by IL-1β TNF-α and diclofenac. IL-1β was the most effective stimulus, and had synergistic effect with TNF-α or diclifenac. MMP-2 activity was not affected. Doxcycline, TGF-β1 and dexamethasone could depress the activities of MMP-9 and MMP-2, and antagonize the enhancing effect of IL-1β TNF-α or diclofenac. Conclusion: IL-1β and TNF-α may play important roles degrading OA cartilage, while TGF-β1 and doxycycline may be protective factors.

  17. Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells Contribute to Chondrogenesis in Coculture with Chondrocytes

    Directory of Open Access Journals (Sweden)

    Xingfu Li

    2016-01-01

    Full Text Available Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs have been shown as the most potential stem cell source for articular cartilage repair. In this study, we aimed to develop a method for long-term coculture of human articular chondrocytes (hACs and hUCB-MSCs at low density in vitro to determine if the low density of hACs could enhance the hUCB-MSC chondrogenic differentiation as well as to determine the optimal ratio of the two cell types. Also, we compared the difference between direct coculture and indirect coculture at low density. Monolayer cultures of hUCB-MSCs and hACs were investigated at different ratios, at direct cell-cell contact groups for 21 days. Compared to direct coculture, hUCB-MSCs and hACs indirect contact culture significantly increased type II collagen (COL2 and decreased type I collagen (COL1 protein expression levels. SRY-box 9 (SOX9 mRNA levels and protein expression were highest in indirect coculture. Overall, these results indicate that low density direct coculture induces fibrocartilage. However, indirect coculture in conditioned chondrocyte cell culture medium can increase expression of chondrogenic markers and induce hUCB-MSCs differentiation into mature chondrocytes. This work demonstrates that it is possible to promote chondrogenesis of hUCB-MSCs in combination with hACs, further supporting the concept of novel coculture strategies for tissue engineering.

  18. Enhanced redifferentiation of chondrocytes on microperiodic silk/gelatin scaffolds: toward tailor-made tissue engineering.

    Science.gov (United States)

    Das, Sanskrita; Pati, Falguni; Chameettachal, Shibu; Pahwa, Shikha; Ray, Alok R; Dhara, Santanu; Ghosh, Sourabh

    2013-02-11

    Direct-write assembly allows rapid fabrication of complex three-dimensional (3D) architectures, such as scaffolds simulating anatomical shapes, avoiding the need for expensive lithographic masks. However, proper selection of polymeric ink composition and tailor-made viscoelastic properties are critically important for smooth deposition of ink and shape retention. Deposition of only silk solution leads to frequent clogging due to shear-induced β-sheet crystallization, whereas optimized viscoelastic property of silk-gelatin blends facilitate the flow of these blends through microcapillary nozzles of varying diameter. This study demonstrates that induction of controlled changes in scaffold surface chemistry, by optimizing silk-gelatin ratio, can govern cell proliferation and maintenance of chondrocyte morphology. Microperiodic silk-gelatin scaffolds can influence postexpansion redifferentiation of goat chondrocytes by enhancing Sox-9 gene expression, aggregation, and driving cartilage matrix production, as evidenced by upregulation of collagen type II and aggrecan expression. The strategy for optimizing redifferentiation of chondrocytes can offer valuable consideration in scaffold-based cartilage repair strategies.

  19. Catabolic effects of FGF-1 on chondrocytes and its possible role in osteoarthritis.

    Science.gov (United States)

    El-Seoudi, Abdellatif; El Kader, Tarek Abd; Nishida, Takashi; Eguchi, Takanori; Aoyama, Eriko; Takigawa, Masaharu; Kubota, Satoshi

    2017-03-25

    Fibroblast growth factor 1 (FGF-1) is a classical member of the FGF family and is produced by chondrocytes cultured from osteoarthritic patients. Also, this growth factor was shown to bind to CCN family protein 2 (CCN2), which regenerates damaged articular cartilage and counteracts osteoarthritis (OA) in an animal model. However, the pathophysiological role of FGF-1 in cartilage has not been well investigated. In this study, we evaluated the effects of FGF-1 in vitro and its production in vivo by use of an OA model. Treatment of human chondrocytic cells with FGF-1 resulted in marked repression of genes for cartilaginous extracellular matrix components, whereas it strongly induced matrix metalloproteinase 13 (MMP-13), representing its catabolic effects on cartilage. Interestingly, expression of the CCN2 gene was dramatically repressed by FGF-1, which repression eventually caused the reduced production of CCN2 protein from the chondrocytic cells. The results of a reporter gene assay revealed that this repression could be ascribed, at least in part, to transcriptional regulation. In contrast, the gene expression of FGF-1 was enhanced by exogenous FGF-1, indicating a positive feedback system in these cells. Of note, induction of FGF-1 was observed in the articular cartilage of a rat OA model. These results collectively indicate a pathological role of FGF-1 in OA development, which includes an insufficient cartilage regeneration response caused by CCN2 down regulation.

  20. Andrographolide enhances proliferation and prevents dedifferentiation of rabbit articular chondrocytes: an in vitro study.

    Science.gov (United States)

    Luo, Li-Ke; Wei, Qing-Jun; Liu, Lei; Zheng, Li; Zhao, Jin-Min

    2015-01-01

    As the main active constituent of Andrographis paniculata that was applied in treatment of many diseases including inflammation in ancient China, andrographolide (ANDRO) was found to facilitate reduction of edema and analgesia in arthritis. This suggested that ANDRO may be promising anti-inflammatory agent to relieve destruction and degeneration of cartilage after inflammation. In this study, the effect of ANDRO on rabbit articular chondrocytes in vitro was investigated. Results showed that not more than 8 μM ANDRO did no harm to chondrocytes (P ANDRO groups comparing to the control (P ANDRO could promote expression of aggrecan, collagen II, and Sox9 genes while downregulating expression of collagen I gene (P 0.05). The viability assay, hematoxylin-eosin, safranin O, and immunohistochemical staining also showed better performances in ANDRO groups. As to the doses, 3 μM ANDRO showed the best performance. The results indicate that ANDRO can accelerate proliferation of rabbit articular chondrocytes in vitro and meanwhile maintain the phenotype, which may provide valuable references for further exploration on arthritis.

  1. Effect of Carnosine in Experimental Arthritis and on Primary Culture Chondrocytes

    Directory of Open Access Journals (Sweden)

    S. Ponist

    2016-01-01

    Full Text Available Carnosine’s (CARN anti-inflammatory potential in autoimmune diseases has been but scarcely investigated as yet. The aim of this study was to evaluate the therapeutic potential of CARN in rat adjuvant arthritis, in the model of carrageenan induced hind paw edema (CARA, and also in primary culture of chondrocytes under H2O2 injury. The experiments were done on healthy animals, arthritic animals, and arthritic animals with oral administration of CARN in a daily dose of 150 mg/kg b.w. during 28 days as well as animals with CARA treated by a single administration of CARN in the same dose. CARN beneficially affected hind paw volume and changes in body weight on day 14 and reduced hind paw swelling in CARA. Markers of oxidative stress in plasma and brain (malondialdehyde, 4-hydroxynonenal, protein carbonyls, and lag time of lipid peroxidation and also activity of gamma-glutamyltransferase were significantly corrected by CARN. CARN also reduced IL-1alpha in plasma. Suppression of intracellular oxidant levels was also observed in chondrocytes pretreated with CARN. Our results obtained on two animal models showed that CARN has systemic anti-inflammatory activity and protected rat brain and chondrocytes from oxidative stress. This finding suggests that CARN might be beneficial for treatment of arthritic diseases.

  2. MicroRNA-181b regulates articular chondrocytes differentiation and cartilage integrity.

    Science.gov (United States)

    Song, Jinsoo; Lee, Myeungsu; Kim, Dongkyun; Han, Jiyeon; Chun, Churl-Hong; Jin, Eun-Jung

    2013-02-08

    MicroRNAs are endogenous gene regulators that have been implicated in various developmental and pathological processes. However, the precise identities and functions of the miRNAs involved in cartilage development are not yet well understood. Here, we report that miR-181b regulates chondrocyte differentiation and maintains cartilage integrity, and is thus a potent therapeutic target. MiR-181b was significantly down-regulated during chondrogenic differentiation of TGF-β3-stimulated limb mesenchymal cells, but it was significantly up-regulated in osteoarthritic chondrocytes isolated from the cartilage of osteoarthritis patients. The use of a mimic or an inhibitor to alter miR-181b levels in chondroblasts and articular chondrocytes showed that attenuation of miR-181b reduced MMP-13 expression while inducing type II collagen expression. Furthermore, over-expression of anti-miR-181b significantly reduced the cartilage destruction caused by DMM surgery in mice. In sum, our data suggest that miR-181b is a negative regulator of cartilage development, and that inhibition of miR-181b could be an effective therapeutic strategy for cartilage-related disease.

  3. Identification of a novel population of human cord blood cells with hematopoietic and chondrocytic potential

    Institute of Scientific and Technical Information of China (English)

    Karen E JAY; Anne ROULEAU; T Michael UNDERHILL; Mickie BHATIA

    2004-01-01

    With the exception of mature erythrocytes, cells within the human hematopoietic system are characterized by the cell surface expression of the pan-leukocyte receptor CD45. Here, we identify a novel subset among mononuclear cord blood cells depleted of lineage commitment markers (Lin-) that are devoid of CD45 expression. Surprisingly, functional examination of Lin-CD45- cells also lacking cell surface CD34 revealed they were capable of multipotential hematopoietic progenitor capacity. Co-culture with mouse embryonic limb bud cells demonstrated that Lin-CD45-CD34- cells were capable of contributing to cartilage nodules and differentiating into human chondrocytes. BMP-4, a mesodermal factor known to promote chondrogenesis, significantly augmented Lin-CD45-CD34- differentiation into chondrocytes.Moreover, unlike CD34+ human hematopoietic stem cells, Lin-CD45-CD34- cells were unable to proliferate or survive in liquid cultures, whereas single Lin-CD45-CD34- cells were able to chimerize the inner cell mass (ICM) of murine blastocysts and proliferate in this embryonic environment. Our study identifies a novel population of Lin-CD45-CD34-cells capable of commitment into both hematopoietic and chondrocytic lineages, suggesting that human cord blood may provide a more ubiquitous source of tissue with broader developmental potential than previously appreciated.

  4. Vanadate proliferative and anti-mineralogenic effects are mediated by MAPK and PI-3K/Ras/Erk pathways in a fish chondrocyte cell line.

    Science.gov (United States)

    Tiago, Daniel M; Cancela, M Leonor; Aureliano, Manuel; Laizé, Vincent

    2008-04-16

    We recently reported proliferative and anti-mineralogenic effects of vanadate on fish chondrocytes and here we investigate the signalling pathways associated with these effects. Our data show that vanadate stimulates chondrocyte proliferation through the MAPK pathway, using signalling mechanisms similar to those used by IGF-1, while it inhibits chondrocyte differentiation/mineralization through a putative PI-3K/Ras/Erk signalling, a pathway shared with insulin. Our data also suggest that vanadate impairs ECM mineralization not only by interfering with regulatory pathways but also by inhibiting enzymatic activity of ALP. Finally, this work provides additional evidence for the conservation, throughout evolution, of mechanisms regulating chondrocyte proliferation and differentiation.

  5. Visualization of living terminal hypertrophic chondrocytes of growth plate cartilage in situ by differential interference contrast microscopy and time-lapse cinematography.

    Science.gov (United States)

    Farnum, C E; Turgai, J; Wilsman, N J

    1990-09-01

    The functional unit within the growth plate consists of a column of chondrocytes that passes through a sequence of phases including proliferation, hypertrophy, and death. It is important to our understanding of the biology of the growth plate to determine if distal hypertrophic cells are viable, highly differentiated cells with the potential of actively controlling terminal events of endochondral ossification prior to their death at the chondro-osseous junction. This study for the first time reports on the visualization of living hypertrophic chondrocytes in situ, including the terminal hypertrophic chondrocyte. Chondrocytes in growth plate explants are visualized using rectified differential interference contrast microscopy. We record and measure, using time-lapse cinematography, the rate of movement of subcellular organelles at the limit of resolution of this light microscopy system. Control experiments to assess viability of hypertrophic chondrocytes include coincubating organ cultures with the intravital dye fluorescein diacetate to assess the integrity of the plasma membrane and cytoplasmic esterases. In this system, all hypertrophic chondrocytes, including the very terminal chondrocyte, exist as rounded, fully hydrated cells. By the criteria of intravital dye staining and organelle movement, distal hypertrophic chondrocytes are identical to chondrocytes in the proliferative and early hypertrophic cell zones.

  6. STUDY OF GRAFT SITES IN EPOXY GRAFT COPOLYMERS BY QUANTUM CHEMISTRY CALCULATIONS

    Institute of Scientific and Technical Information of China (English)

    Song Chen; Xiao-yu Li

    2009-01-01

    Exploration and characterization of grafting productions by experimental methods are often cumbersome or sometimes impossible. Therefore, quantum chemistry calculations were performed to characterize the graft sites of epoxy resin. According to the Gibbs free energy criterion of the second law of thermodynamic, the reported graft sites were confirmed, and more important, some unreported graft sites were found. In addition, method of increasing the number of graft sites was studied in this article.

  7. Polyether-polyester graft copolymer

    Science.gov (United States)

    Bell, Vernon L. (Inventor)

    1987-01-01

    Described is a polyether graft polymer having improved solvent resistance and crystalline thermally reversible crosslinks. The copolymer is prepared by a novel process of anionic copolymerization. These polymers exhibit good solvent resistance and are well suited for aircraft parts. Previous aromatic polyethers, also known as polyphenylene oxides, have certain deficiencies which detract from their usefulness. These commercial polymers are often soluble in common solvents including the halocarbon and aromatic hydrocarbon types of paint thinners and removers. This limitation prevents the use of these polyethers in structural articles requiring frequent painting. In addition, the most popular commercially available polyether is a very high melting plastic. This makes it considerably more difficult to fabricate finished parts from this material. These problems are solved by providing an aromatic polyether graft copolymer with improved solvent resistance and crystalline thermally reversible crosslinks. The graft copolymer is formed by converting the carboxyl groups of a carboxylated polyphenylene oxide polymer to ionic carbonyl groups in a suitable solvent, reacting pivalolactone with the dissolved polymer, and adding acid to the solution to produce the graft copolymer.

  8. TNF Accelerates Death of Mandibular Condyle Chondrocytes in Rats with Biomechanical Stimulation-Induced Temporomandibular Joint Disease

    Science.gov (United States)

    Zhang, Hongyun; Zhang, Jing; Jing, Lei; Liao, Lifan; Wang, Meiqing

    2015-01-01

    Objective To determine if temporomandibular joint chondrocyte apoptosis is induced in rats with dental biomechanical stimulation and what a role TNF takes. Methods Thirty-two rats were divided into 4 groups (n = 8/group) and exposed to incisor mal-occlusion induced by unilateral anterior crossbite biomechanical stimulation. Two groups were sampled at 2 or 4 weeks. The other two groups were treated with local injections of a TNF inhibitor or PBS into the temporomandibular joints area at 2 weeks and then sampled at 4 weeks. Twenty-four rats either served as unilateral anterior crossbite mock operation controls (n = 8/group) with sampling at 2 or 4 weeks or received a local injection of the TNF inhibitor at 2 weeks with sampling at 4 weeks. Chondrocytes were isolated from the temporomandibular joints of 6 additional rats and treated with TNF in vitro. Joint samples were assessed using Hematoxylin&eosin, Safranin O, TUNEL and immunohistochemistry staining, real-time PCR, fluorogenic activity assays and Western blot analyses. The isolated chondrocytes were also analyzed by flow cytometry. Results Unilateral anterior crossbite stimulation led to temporomandibular joint cartilage degradation, associated with an increase in TUNEL-positive chondrocytes number, caspase-9 expression levels, and the release of cytochrome c from mitochondria at 2 weeks without changes in TNF and caspase-8 levels until after 4 weeks. TNF stimulated apoptosis of the isolated chondrocytes and up-regulated caspase-8 expression, but did not change caspase-9 expression levels. Local injection of TNF inhibitor down-regulated caspase-8 expression and reduced TUNEL-positive cell number, but did not reverse cartilage thickness reduction, caspase-9 up-regulation or cytochrome c release. Conclusions Unilateral anterior crossbite stimulation induces mitochondrion-mediated apoptosis of articular chondrocytes. TNF accelerated the unilateral anterior crossbite induced chondrocytes apoptosis via death

  9. CD44 knock-down in bovine and human chondrocytes results in release of bound HYAL2

    Science.gov (United States)

    Hida, Daisuke; Danielson, Ben T.; Knudson, Cheryl B.; Knudson, Warren

    2015-01-01

    CD44 shedding occurs in osteoarthritic chondrocytes. Previous work of others has suggested that the hyaluronidase isoform HYAL2 has the capacity to bind to CD44, a binding that may itself induce CD44 cleavage. Experiments were developed to elucidate whether chondrocyte HYAL2: (1) was exposed on the extracellular plasma membrane of chondrocytes, (2) bound to CD44, (3) underwent shedding together with CD44 and lastly, (4) exhibited hyaluronidase activity within a near-neutral pH range. Enhancing CD44 shedding by IL-1β resulted in a proportional increase in HYAL2 released from human and bovine chondrocytes into the medium. CD44 knockdown by siRNA also resulted in increased accumulation of HYAL2 in the media of chondrocytes. By hyaluronan zymography only activity at pH 3.7 was observed and this activity was reduced by pre-treatment of chondrocytes with trypsin. CD44 and HYAL2 were found to co-immunoprecipitate, and to co-localize within intracellular vesicles and at the plasma membrane. Degradation of hyaluronan was visualized by agarose gel electrophoresis. With this approach, hyaluronidase activity could be observed at pH 4.8 under assay conditions in which CD44 and HYAL2 binding remained intact; additionally, weak hyaluronidase activity could be observed at pH 6.8 under these conditions. This study suggests that CD44 and HYAL2 are bound at the surface of chondrocytes. The release of HYAL2 when CD44 is shed could provide a mechanism for weak hyaluronidase activity to occur within the more distant extracellular matrix of cartilage. PMID:25864644

  10. TNF Accelerates Death of Mandibular Condyle Chondrocytes in Rats with Biomechanical Stimulation-Induced Temporomandibular Joint Disease.

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    Hongxu Yang

    Full Text Available To determine if temporomandibular joint chondrocyte apoptosis is induced in rats with dental biomechanical stimulation and what a role TNF takes.Thirty-two rats were divided into 4 groups (n = 8/group and exposed to incisor mal-occlusion induced by unilateral anterior crossbite biomechanical stimulation. Two groups were sampled at 2 or 4 weeks. The other two groups were treated with local injections of a TNF inhibitor or PBS into the temporomandibular joints area at 2 weeks and then sampled at 4 weeks. Twenty-four rats either served as unilateral anterior crossbite mock operation controls (n = 8/group with sampling at 2 or 4 weeks or received a local injection of the TNF inhibitor at 2 weeks with sampling at 4 weeks. Chondrocytes were isolated from the temporomandibular joints of 6 additional rats and treated with TNF in vitro. Joint samples were assessed using Hematoxylin&eosin, Safranin O, TUNEL and immunohistochemistry staining, real-time PCR, fluorogenic activity assays and Western blot analyses. The isolated chondrocytes were also analyzed by flow cytometry.Unilateral anterior crossbite stimulation led to temporomandibular joint cartilage degradation, associated with an increase in TUNEL-positive chondrocytes number, caspase-9 expression levels, and the release of cytochrome c from mitochondria at 2 weeks without changes in TNF and caspase-8 levels until after 4 weeks. TNF stimulated apoptosis of the isolated chondrocytes and up-regulated caspase-8 expression, but did not change caspase-9 expression levels. Local injection of TNF inhibitor down-regulated caspase-8 expression and reduced TUNEL-positive cell number, but did not reverse cartilage thickness reduction, caspase-9 up-regulation or cytochrome c release.Unilateral anterior crossbite stimulation induces mitochondrion-mediated apoptosis of articular chondrocytes. TNF accelerated the unilateral anterior crossbite induced chondrocytes apoptosis via death-receptor pathway. However, anti

  11. JNK phosphorylation promotes degeneration of cervical endplate chondrocytes through down-regulation of the expression of ANK in humans

    Institute of Scientific and Technical Information of China (English)

    XU Hong-guang; SONG Jun-xing; CHENG Jia-feng; ZHANG Ping-Zhi; WANG Hong; LIU Ping; L(U) Kun

    2013-01-01

    Background C-Jun N-terminal kinase (JNK) signaling pathway and ankylosis gene (ANK) play a critical role in endplate chondrocytes degeneration.The purpose of this study was to investigate whether the expression levels of ANK was associated with the activation of JNK.Methods Cartilage endplates of 49 patients were divided into the control group (n=19) and the experimental group (n=30).The patients in the control group were graded 0 and those in the experimental group were graded Ⅰ-Ⅲ according to Miller's classification.Endplate chondrocytes were isolated by enzyme digestion and cultured in vitro.The inverted phase contrast microscope,teluidine blue staining,HE staining,real time RT-PCR,and MTT were used to observe morphological appearances,biological characteristics,and growth curve of endplate chondrocytes from the cartilage endplate of the two groups.Real time RT-PCR and Western blotting were used to analyze the mRNA and protein expression levels of associated factors in the degeneration process in the cultured endplate chondrocytes with or without subjected SP600125.Results The expression levels of type Ⅱ collagen,aggrecan,and ANK in endplate chondrocytes of experimental group were lower than that of control group and phosphorylation level of JNK in the experimental group which was higher than that in the control group.Application of JNK phosphorylation inhibitor to degeneration chondrocytes resulted in a marked decrease in the phosphorylation level of JNK and a significant increase in the expression levels of type Ⅱ collagen,aggrecan,and ANK.Conclusion The degeneration of the human cervical endplate chondrocytes might be promoted by JNK phosphorylation by down-regulating the expression of ANK

  12. Nicotine acts on growth plate chondrocytes to delay skeletal growth through the alpha7 neuronal nicotinic acetylcholine receptor.

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    Atsuo Kawakita

    Full Text Available BACKGROUND: Cigarette smoking adversely affects endochondral ossification during the course of skeletal growth. Among a plethora of cigarette chemicals, nicotine is one of the primary candidate compounds responsible for the cause of smoking-induced delayed skeletal growth. However, the possible mechanism of delayed skeletal growth caused by nicotine remains unclarified. In the last decade, localization of neuronal nicotinic acetylcholine receptor (nAChR, a specific receptor of nicotine, has been widely detected in non-excitable cells. Therefore, we hypothesized that nicotine affect growth plate chondrocytes directly and specifically through nAChR to delay skeletal growth. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the effect of nicotine on human growth plate chondrocytes, a major component of endochondral ossification. The chondrocytes were derived from extra human fingers. Nicotine inhibited matrix synthesis and hypertrophic differentiation in human growth plate chondrocytes in suspension culture in a concentration-dependent manner. Both human and murine growth plate chondrocytes expressed alpha7 nAChR, which constitutes functional homopentameric receptors. Methyllycaconitine (MLA, a specific antagonist of alpha7 nAChR, reversed the inhibition of matrix synthesis and functional calcium signal by nicotine in human growth plate chondrocytes in vitro. To study the effect of nicotine on growth plate in vivo, ovulation-controlled pregnant alpha7 nAChR +/- mice were given drinking water with or without nicotine during pregnancy, and skeletal growth of their fetuses was observed. Maternal nicotine exposure resulted in delayed skeletal growth of alpha7 nAChR +/+ fetuses but not in alpha7 nAChR -/- fetuses, implying that skeletal growth retardation by nicotine is specifically mediated via fetal alpha7 nAChR. CONCLUSIONS/SIGNIFICANCE: These results suggest that nicotine, from cigarette smoking, acts directly on growth plate chondrocytes to decrease

  13. Electrospun gelatin/polycaprolactone nanofibrous membranes combined with a coculture of bone marrow stromal cells and chondrocytes for cartilage engineering

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    He X

    2015-03-01

    Full Text Available Xiaomin He,1,* Bei Feng,1,2,* Chuanpei Huang,1 Hao Wang,1 Yang Ge,1 Renjie Hu,1 Meng Yin,1 Zhiwei Xu,1 Wei Wang,1 Wei Fu,1,2 Jinghao Zheng1 1Department of Pediatric Cardiothoracic Surgery, 2Institute of Pediatric Translational Medicine, Shanghai Children’s Medical Center School of Medicine, Shanghai Jiao Tong University, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: Electrospinning has recently received considerable attention, showing notable potential as a novel method of scaffold fabrication for cartilage engineering. The aim of this study was to use a coculture strategy of chondrocytes combined with electrospun gelatin/polycaprolactone (GT/PCL membranes, instead of pure chondrocytes, to evaluate the formation of cartilaginous tissue. We prepared the GT/PCL membranes, seeded bone marrow stromal cell (BMSC/chondrocyte cocultures (75% BMSCs and 25% chondrocytes in a sandwich model in vitro, and then implanted the constructs subcutaneously into nude mice for 12 weeks. Gross observation, histological and immunohistological evaluation, glycosaminoglycan analyses, Young’s modulus measurement, and immunofluorescence staining were performed postimplantation. We found that the coculture group formed mature cartilage-like tissue, with no statistically significant difference from the chondrocyte group, and labeled BMSCs could differentiate into chondrocyte-like cells under the chondrogenic niche of chondrocytes. This entire strategy indicates that GT/PCL membranes are also a suitable scaffold for stem cell-based cartilage engineering and may provide a potentially clinically feasible approach for cartilage repairs. Keywords: electrospinning, nanocomposite, cartilage tissue engineering, nanomaterials, stem cells

  14. Determinants of microstructural load transfer in cartilage tissue from chondrocyte culture

    Science.gov (United States)

    Fedewa, Michelle Marie

    2000-10-01

    The goals of this research were to (i) develop a tissue model system for studying the microstructure of matrix produced by chondrocytes, (ii) characterize the biochemical and mechanical properties of the chondrocyte culture tissue, (iii) evaluate the response of the chondrocyte culture tissue to various stimulants (retinoic acid, interleukin-1beta, and xyloside), (iv) investigate the roles of proteoglycan and collagen in the tearing and tensile properties of a chondrocyte culture tissue, and (v) develop a finite element model of the chondrocyte culture tissue microstructure to study its tensile pre-failure properties. The roles of proteoglycan and collagen were explored by experimentation using a cultured cartilage tissue, and by development of a theoretical finite element model which related the cartilage tissue microstructure to its macroscopic properties. Tear and tensile testing was performed. Failure testing is valuable because it is known that cracks exist and propagate from the cartilage surface in osteoarthritic joints. It was found that collagen was important for providing the material stiffness of the cultured tissue, and that both collagen and proteoglycan were important for providing the tear toughness of the tissue. It was also found that as the collagen density or collagen material stiffness increased, the material stiffness of the cultured tissue increased, and as the proteoglycan or collagen densities increased, the tear toughness of the tissue increased. A three-dimensional finite element microstructural model of cartilage was developed, consisting of linear elastic collagen fibrils embedded in a linear viscoelastic proteoglycan solid matrix. Fluid flow in the cartilage matrix was not included in this model. Viscoelastic time dependent behavior was an appropriate model for the cartilage. The results of this model were comparable to the experimental results, as well as to past continuum models of cartilage. Collagen and proteoglycan material moduli

  15. Evidence for regulated interleukin-4 expression in chondrocyte-scaffolds under in vitro inflammatory conditions.

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    Muhammad Farooq Rai

    Full Text Available OBJECTIVE: To elucidate the anti-inflammatory and anabolic effects of regulated expression of IL-4 in chondrocyte-scaffolds under in vitro inflammatory conditions. METHODS: Mature articular chondrocytes from dogs (n = 3 were conditioned through transient transfection using pcDNA3.1.cIL-4 (constitutive or pCOX-2.cIL-4 (cytokine-responsive plasmids. Conditioned cells were seeded in alginate microspheres and rat-tail collagen type I matrix (CaReS® to generate two types of tissue-engineered 3-dimensional scaffolds. Inflammatory arthritis was simulated in the packed chondrocytes through exogenous addition of recombinant canine (rc IL-1β (100 ng/ml plus rcTNFα (50 ng/ml in culture media for 96 hours. Harvested cells and culture media were analyzed by various assays to monitor the anti-inflammatory and regenerative (anabolic properties of cIL-4. RESULTS: cIL-4 was expressed from COX-2 promoter exclusively on the addition of rcIL-1β and rcTNFα while its expression from CMV promoter was constitutive. The expressed cIL-4 downregulated the mRNA expression of IL-1β, TNFα, IL-6, iNOS and COX-2 in the cells and inhibited the production of NO and PGE(2 in culture media. At the same time, it up-regulated the expression of IGF-1, IL-1ra, COL2a1 and aggrecan in conditioned chondrocytes in both scaffolds along with a diminished release of total collagen and sGAG into the culture media. An increased amount of cIL-4 protein was detected both in chondrocyte cell lysate and in concentrated culture media. Neutralizing anti-cIL-4 antibody assay confirmed that the anti-inflammatory and regenerative effects seen are exclusively driven by cIL-4. There was a restricted expression of IL-4 under COX-2 promoter possibly due to negative feedback loop while it was over-expressed under CMV promoter (undesirable. Furthermore, the anti-inflammatory /anabolic outcomes from both scaffolds were reproducible and the therapeutic effects of cIL-4 were both scaffold- and

  16. Effect of the disruption of three cytoskeleton components on chondrocyte metabolism in rabbit knee cartilage

    Institute of Scientific and Technical Information of China (English)

    Duan Wangping; Wei Lei; Cao Xiaoming; Guo Heng; Wang Lei; Hao Yongzhuang; Wei Xiaochun

    2014-01-01

    Background Chondrocytes' phenotype and biosynthesis of matrix are dependent on having an intact cytoskeletal structure.Microfilaments,microtubules,and intermediate filaments are three important components of the cytoskeletal structure of chondrocytes.The aims of this study were to determine and compare the effects of the disruption of these three cytoskeletal elements on the apoptosis and matrix synthesis by rabbit knee chondrocytes in vitro.Methods Chondrocytes were isolated from full-thickness knee cartilage of two-month-old rabbits using enzymatic methods (n=24).The isolated cells were stabilized for three days and then exposed to low,medium,and high doses of chemical agents that disrupt the three principal cytoskeletal elements of interest:colchicine for microtubules,acrylamide for intermediate filaments,and cytochalasin D for actin microfilaments.A group of control cells were treated with carrier.Early apoptosis was assessed using the Annexin-FITC binding assay by flow cytometry on days 1 and 2 after exposure to the disrupting chemical agents.The components and distribution of the cytoskeleton within the cells were analyzed by laser scanning confocal microscopy (LSCM) with immunofluorescence staining on day 3.The mRNA levels of aggrecan (AGG) and type Ⅱ collagen (Col-2) and their levels in culture medium were analyzed using real-time PCR and enzymelinked immunosorbent serologic assay (ELISA) on days 3,6,and 9.Results In the initial drug-dose-response study,there was no significant difference in the vitality of cells treated with 0.1 μmol/L colchicine,2.5 mmol/L acrylamide,and 10 μg/L cytochalasin D for two days when compared with the control group of cells.The concentrations of colchicine and acrylamide treatment selected above significantly decreased the number of viable cells over the nine-day culture and disrupted significantly more cell nuclei.Real-time PCR and ELISA results showed that the mRNA levels and medium concentrations of AGG and Col-2 were

  17. Autologous chondrocyte implantation in the knee: systematic review and economic evaluation.

    Science.gov (United States)

    Mistry, Hema; Connock, Martin; Pink, Joshua; Shyangdan, Deepson; Clar, Christine; Royle, Pamela; Court, Rachel; Biant, Leela C; Metcalfe, Andrew; Waugh, Norman

    2017-01-01

    BACKGROUND The surfaces of the bones in the knee are covered with articular cartilage, a rubber-like substance that is very smooth, allowing frictionless movement in the joint and acting as a shock absorber. The cells that form the cartilage are called chondrocytes. Natural cartilage is called hyaline cartilage. Articular cartilage has very little capacity for self-repair, so damage may be permanent. Various methods have been used to try to repair cartilage. Autologous chondrocyte implantation (ACI) involves laboratory culture of cartilage-producing cells from the knee and then implanting them into the chondral defect. OBJECTIVE To assess the clinical effectiveness and cost-effectiveness of ACI in chondral defects in the knee, compared with microfracture (MF). DATA SOURCES A broad search was done in MEDLINE, EMBASE, The Cochrane Library, NHS Economic Evaluation Database and Web of Science, for studies published since the last Health Technology Assessment review. REVIEW METHODS Systematic review of recent reviews, trials, long-term observational studies and economic evaluations of the use of ACI and MF for repairing symptomatic articular cartilage defects of the knee. A new economic model was constructed. Submissions from two manufacturers and the ACTIVE (Autologous Chondrocyte Transplantation/Implantation Versus Existing Treatment) trial group were reviewed. Survival analysis was based on long-term observational studies. RESULTS Four randomised controlled trials (RCTs) published since the last appraisal provided evidence on the efficacy of ACI. The SUMMIT (Superiority of Matrix-induced autologous chondrocyte implant versus Microfracture for Treatment of symptomatic articular cartilage defects) trial compared matrix-applied chondrocyte implantation (MACI(®)) against MF. The TIG/ACT/01/2000 (TIG/ACT) trial compared ACI with characterised chondrocytes against MF. The ACTIVE trial compared several forms of ACI against standard treatments, mainly MF. In the SUMMIT

  18. Posterior repair with perforated porcine dermal graft

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    G. Bernard Taylor

    2008-02-01

    Full Text Available OBJECTIVE: To compare postoperative vaginal incision separation and healing in patients undergoing posterior repair with perforated porcine dermal grafts with those that received grafts without perforations. Secondarily, the tensile properties of the perforated and non-perforated grafts were measured and compared. MATERIALS AND METHODS: This was a non-randomized retrospective cohort analysis of women with stage II or greater rectoceles who underwent posterior repair with perforated and non-perforated porcine dermal grafts (PelvicolTM CR Bard Covington, GA USA. The incidence of postoperative vaginal incision separation (dehiscence was compared. A secondary analysis to assess graft tensile strength, suture pull out strength, and flexibility after perforation was performed using standard test method TM 0133 and ASTM bending and resistance protocols. RESULTS: Seventeen percent of patients (21/127 who received grafts without perforations developed vaginal incision dehiscence compared to 7% (5/71 of patients who received perforated grafts (p = 0.078. Four patients with vaginal incision dehiscence with non-perforated grafts required surgical revision to facilitate healing. Neither tensile strength or suture pull out strength were significantly different between perforated and non-perforated grafts (p = 0.81, p = 0.29, respectively. There was no difference in the flexibility of the two grafts (p = 0.20. CONCLUSION: Perforated porcine dermal grafts retain their tensile properties and are associated with fewer vaginal incision dehiscences.

  19. Starch Modification by Graft Copolymerization of Acrylonitrile

    Institute of Scientific and Technical Information of China (English)

    刘瑞贤; 李莉; 茹宗玲; 张黎明; 高建平; 田汝川

    2003-01-01

    The graft copolymerization of acrylonitrile (AN) onto starch under the initiation of potassium permanganate was investigated. The effect of various reaction conditions on the graft copolymerization was studied. The relationships between the grafting rate and the initiator concentration of potassium permanganate, monomer acrylonitrile and backbone starch, as well as reaction temperature were established. The oxidation reaction of starch with manganic ions and valence changes of manganic ions during the graft copolymerization were discussed. The results show that manganic ion Mn7+ underwent a series of valence changes during the graft copolymerization: Mn(Ⅶ) → Mn(Ⅳ) → Mn(Ⅲ) → Mn(Ⅱ). The grafting rate of the graft copolymerization of acrylonitrile onto starch is also given.

  20. Dynamics of bone graft healing around implants

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    Narayan Venkataraman

    2015-01-01

    A few questions arise pertaining to the use of bone grafts along with implants are whether these are successful in approximation with implant. Do they accelerate bone regeneration? Are all defects ultimately regenerated with new viable bone? Is the bone graft completely resorbed or integrated in new bone? Does the implant surface characteristic positively affect osseointegration when used with a bone graft? What type of graft and implant surface can be used that will have a positive effect on the healing type and time? Finally, what are the dynamics of bone graft healing around an implant? This review discusses the cellular and molecular mechanisms of bone graft healing in general and in vicinity of another foreign, avascular body, namely the implant surface, and further, the role of bone grafts in osseointegration and/or clinical success of the implants.

  1. Radiation-induced grafting of diallyldimethylammonium chloride onto acrylic acid grafted polyethylene

    Science.gov (United States)

    Francis, Sanju; Dhanawade, B. R.; Mitra, D.; Varshney, Lalit; Sabharwal, Sunil

    2009-01-01

    Diallyldimethylammonium chloride (DADMAC) was grafted onto polyethylene (PE) films by a double grafting procedure. The PE film was initially modified by grafting acrylic acid (AA), through a mutual irradiation method. AA-g-PE film, thus obtained was subjected to subsequent radiation grafting reaction of DADMAC, to give a DADMAC-g-AA-g-PE film having a comb-type structure. The influence of different conditions, such as the extent of AA grafting, DADMAC concentration, absorbed dose and dose rate, on the grafting yield of DADMAC was investigated. A maximum DADMAC grafting of 30% was achieved. The equilibrium degree of swelling (EDS) of the grafted films were gravimetrically determined. TGA and FT-IR techniques were employed to characterize the grafted PE films.

  2. Radiation-induced grafting of diallyldimethylammonium chloride onto acrylic acid grafted polyethylene

    Energy Technology Data Exchange (ETDEWEB)

    Francis, Sanju [Radiation Technology Development Section, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085 (India)], E-mail: sanju@barc.gov.in; Dhanawade, B.R.; Mitra, D.; Varshney, Lalit; Sabharwal, Sunil [Radiation Technology Development Section, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085 (India)

    2009-01-15

    Diallyldimethylammonium chloride (DADMAC) was grafted onto polyethylene (PE) films by a double grafting procedure. The PE film was initially modified by grafting acrylic acid (AA), through a mutual irradiation method. AA-g-PE film, thus obtained was subjected to subsequent radiation grafting reaction of DADMAC, to give a DADMAC-g-AA-g-PE film having a comb-type structure. The influence of different conditions, such as the extent of AA grafting, DADMAC concentration, absorbed dose and dose rate, on the grafting yield of DADMAC was investigated. A maximum DADMAC grafting of 30% was achieved. The equilibrium degree of swelling (EDS) of the grafted films were gravimetrically determined. TGA and FT-IR techniques were employed to characterize the grafted PE films.

  3. Potential of Raloxifene in reversing osteoarthritis-like alterations in rat chondrocytes: An in vitro model study

    Indian Academy of Sciences (India)

    Aysegul Kavas; Seda Tuncay Cagatay; Sreeparna Banerjee; Dilek Keskin; Aysen Tezcaner

    2013-03-01

    The aim of this study was to investigate the effects of Raloxifene (Ral) on degeneration-related changes in osteoarthritis (OA)-like chondrocytes using two- and three-dimensional models. Five-azacytidine (Aza-C) was used to induce OA-like alterations in rat articular chondrocytes and the model was verified at molecular and macrolevels. Chondrocytes were treated with Ral (1, 5 and 10 M) for 10 days. Caspase-3 activity, gene expressions of aggrecan, collagen II, alkaline phosphatase (ALP), collagen X, matrix metalloproteinases (MMP-13, MMP-3 and MMP-2), and MMP-13, MMP-3 and MMP-2 protein expressions were studied in two-dimensional model. Matrix deposition and mechanical properties of agarose-chondrocyte discs were evaluated in three-dimensional model. One M Ral reduced expression of OA-related genes, decreased apoptosis, and MMP-13 and MMP-3 protein expressions. It also increased aggrecan and collagen II gene expressions relative to untreated OA-like chondrocytes. In three-dimensional model, 1 M Ral treatment resulted in increased collagen deposition and improved mechanical properties, although a significant increase for sGAG was not observed. In summation, 1 M Ral improved matrix-related activities, whereas dose increment reversed these effects except ALP gene expression and sGAG deposition. These results provide evidence that low-dose Ral has the potential to cease or reduce the matrix degeneration in OA.

  4. Lubricin is expressed in chondrocytes derived from osteoarthritic cartilage encapsulated in poly(ethylene glycol diacrylate scaffold

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    G. Musumeci

    2011-09-01

    Full Text Available Osteoarthritis (OA is characterized by degenerative changes within joints that involved quantitative and/or qualitative alterations of cartilage and synovial fluid lubricin, a mucinous glycoprotein secreted by synovial fibroblasts and chondrocytes. Modern therapeutic methods, including tissue-engineering techniques, have been used to treat mechanical damage of the articular cartilage but to date there is no specific and effective treatment. This study aimed at investigating lubricin immunohistochemical expression in cartilage explant from normal and OA patients and in cartilage constructions formed by Poly (ethylene glycol (PEG based hydrogels (PEG-DA encapsulated OA chondrocytes. The expression levels of lubricin were studied by immunohistochemistry: i in tissue explanted from OA and normal human cartilage; ii in chondrocytes encapsulated in hydrogel PEGDA from OA and normal human cartilage. Moreover, immunocytochemical and western blot analysis were performed in monolayer cells from OA and normal cartilage. The results showed an increased expression of lubricin in explanted tissue and in monolayer cells from normal cartilage, and a decreased expression of lubricin in OA cartilage. The chondrocytes from OA cartilage after 5 weeks of culture in hydrogels (PEGDA showed an increased expression of lubricin compared with the control cartilage. The present study demonstrated that OA chondrocytes encapsulated in PEGDA, grown in the scaffold and were able to restore lubricin biosynthesis. Thus our results suggest the possibility of applying autologous cell transplantation in conjunction with scaffold materials for repairing cartilage lesions in patients with OA to reduce at least the progression of the disease.

  5. The effect of oxygen tension on human articular chondrocyte matrix synthesis: integration of experimental and computational approaches.

    Science.gov (United States)

    Li, S; Oreffo, R O C; Sengers, B G; Tare, R S

    2014-09-01

    Significant oxygen gradients occur within tissue engineered cartilaginous constructs. Although oxygen tension is an important limiting parameter in the development of new cartilage matrix, its precise role in matrix formation by chondrocytes remains controversial, primarily due to discrepancies in the experimental setup applied in different studies. In this study, the specific effects of oxygen tension on the synthesis of cartilaginous matrix by human articular chondrocytes were studied using a combined experimental-computational approach in a "scaffold-free" 3D pellet culture model. Key parameters including cellular oxygen uptake rate were determined experimentally and used in conjunction with a mathematical model to estimate oxygen tension profiles in 21-day cartilaginous pellets. A threshold oxygen tension (pO2 ≈ 8% atmospheric pressure) for human articular chondrocytes was estimated from these inferred oxygen profiles and histological analysis of pellet sections. Human articular chondrocytes that experienced oxygen tension below this threshold demonstrated enhanced proteoglycan deposition. Conversely, oxygen tension higher than the threshold favored collagen synthesis. This study has demonstrated a close relationship between oxygen tension and matrix synthesis by human articular chondrocytes in a "scaffold-free" 3D pellet culture model, providing valuable insight into the understanding and optimization of cartilage bioengineering approaches.

  6. Mechanical Forces Induce Changes in VEGF and VEGFR-1/sFlt-1 Expression in Human Chondrocytes

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    Rainer Beckmann

    2014-09-01

    Full Text Available Expression of the pro-angiogenic vascular endothelial growth factor (VEGF stimulates angiogenesis and correlates with the progression of osteoarthritis. Mechanical joint loading seems to contribute to this cartilage pathology. Cyclic equibiaxial strains of 1% to 16% for 12 h, respectively, induced expression of VEGF in human chondrocytes dose- and frequency-dependently. Stretch-mediated VEGF induction was more prominent in the human chondrocyte cell line C-28/I2 than in primary articular chondrocytes. Twelve hours of 8% stretch induced VEGF expression to 175% of unstrained controls for at least 24 h post stretching, in promoter reporter and enzyme-linked immunosorbent assay (ELISA studies. High affinity soluble VEGF-receptor, sVEGFR-1/sFlt-1 was less stretch-inducible than its ligand, VEGF-A, in these cells. ELISA assays demonstrated, for the first time, a stretch-mediated suppression of sVEGFR-1 secretion 24 h after stretching. Overall, strained chondrocytes activate their VEGF expression, but in contrast, strain appears to suppress the secretion of the major VEGF decoy receptor (sVEGFR-1/sFlt-1. The latter may deplete a biologically relevant feedback regulation to inhibit destructive angiogenesis in articular cartilage. Our data suggest that mechanical stretch can induce morphological changes in human chondrocytes in vitro. More importantly, it induces disturbed VEGF signaling, providing a molecular mechanism for a stress-induced increase in angiogenesis in cartilage pathologies.

  7. THE ACTIVATION OF MATRIX METALLOPROTEINASES AND CHONDROCYTE DIFFERENTIATION, WHICH ACCOMPANIES THE INDUCTION OF COLLAGEN DECOMPOSITION UNDER THE ACTION OF COLLAGEN PEPTIDE IN THE CARTILAGE OFHEALTHY INDIVIDUALS

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    Elena Vasil'evna Chetina

    2010-01-01

    Conclusion. This study has shown that the induction of collagenase activity by CB12-2 in the human articular cartilage chondrocytes is attended by terminal differentiation/hypertrophy of these cells. The terminal differentiation of chondrocytes may be one of the mechanisms of chondrolysis in osteoarthrosis since it naturally occurs not only in endochondrial ossification, but also in the development of pathology.

  8. Matrine inhibits IL-1β-induced expression of matrix metalloproteinases by suppressing the activation of MAPK and NF-κB in human chondrocytes in vitro.

    Science.gov (United States)

    Lu, Shijin; Xiao, Xungang; Cheng, Minghua

    2015-01-01

    Interleukin (IL)-1β plays an important role in promoting osteoarthritis (OA) lesions by inducing chondrocytes to secrete matrix metalloproteinases (MMPs), which degrade the extracellular matrix and facilitate chondrocyte apoptosis. Matrine was shown to exert anti-inflammatory effects. However, the role of matrine in OA is still unclear. Therefore, in this study, we investigated the effects of matrine on the expression of MMPs in IL-1β-treated human chondrocytes and the underlying mechanism. The cell viability of chondrocytes was detected by MTT assay. The cell apoptosis of chondrocytes was measured by flow cytometric analysis. The protein production of MMPs was determined by ELISA. The protein expression of phosphorylation of mitogen-activated protein kinases (MAPKs) and the inhibitor of kappaB alpha (IκBα) was determined by Western blot. Matrine significantly inhibited the IL-1β-induced apoptosis in chondrocytes. It also significantly inhibited the IL-1β-induced release of MMP-3 and MMP-13, and increased the production of TIMP-1. Furthermore, matrine inhibits the phosphorylation of p-38, extracellular regulated kinase (ERK), c-Jun-N-terminal kinase (JNK) and IκBα degradation induced by IL-1β in chondrocytes. Taken together, our results show that matrine inhibits IL-1β-induced expression of matrix metalloproteinases by suppressing the activation of MAPK and NF-κB in human chondrocytes in vitro. Therefore,-matrine may be beneficial in the treatment of OA.

  9. Expression Profiling and Functional Implications of a Set of Zinc Finger Proteins, ZNF423, ZNF470, ZNF521, and ZNF780B, in Primary Osteoarthritic Articular Chondrocytes

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    Maria Mesuraca

    2014-01-01

    Full Text Available Articular chondrocytes are responsible for the maintenance of healthy articulations; indeed, dysregulation of their functions, including the production of matrix proteins and matrix-remodeling proteases, may result in fraying of the tissue and development of osteoarthritis (OA. To explore transcriptional mechanisms that contribute to the regulation of chondrocyte homeostasis and may be implicated in OA development, we compared the gene expression profile of a set of zinc finger proteins potentially linked to the control of chondrocyte differentiation and/or functions (ZNF423, ZNF470, ZNF521, and ZNF780B in chondrocytes from patients affected by OA and from subjects not affected by OA. This analysis highlighted a significantly lower expression of the transcript encoding ZNF423 in chondrocytes from OA, particularly in elderly patients. Interestingly, this decrease was mirrored by the similarly reduced expression of PPARγ, a known target of ZNF423 with anti-inflammatory and chondroprotective properties. The ZNF521 mRNA instead was abundant in all primary chondrocytes studied; the RNAi-mediated silencing of this gene significantly altered the COL2A/COL1 expression ratio, associated with the maintenance of the differentiated phenotype, in chondrocytes cultivated in alginate beads. These results suggest a role for ZNF423 and ZNF521 in the regulation of chondrocyte homeostasis and warrant further investigations to elucidate their mechanism of action.

  10. Evaluation of Magnetic Nanoparticle-Labeled Chondrocytes Cultivated on a Type II Collagen–Chitosan/Poly(Lactic-co-Glycolic) Acid Biphasic Scaffold

    OpenAIRE

    Juin-Yih Su; Shi-Hui Chen; Yu-Pin Chen; Wei-Chuan Chen

    2017-01-01

    Chondral or osteochondral defects are still controversial problems in orthopedics. Here, chondrocytes labeled with magnetic nanoparticles were cultivated on a biphasic, type II collagen–chitosan/poly(lactic-co-glycolic acid) scaffold in an attempt to develop cultures with trackable cells exhibiting growth, differentiation, and regeneration. Rabbit chondrocytes were labeled with magnetic nanoparticles and characterized by scanning electron microscopy (SEM), transmission electron (TEM) microsco...

  11. Inducible chondrocyte-specific overexpression of BMP2 in young mice results in severe aggravation of osteophyte formation in experimental OA without altering cartilage damage

    NARCIS (Netherlands)

    Davidson, E.N.; Vitters, E.L.; Bennink, M.B.; Lent, P.L. van; Caam, A.P. van; Blom, A.B.; Berg, W.B. van den; Loo, F.A.J. van de; Kraan, P.M. van der

    2015-01-01

    OBJECTIVES: In osteoarthritis (OA) chondrocytes surrounding lesions express elevated bone morphogenetic protein 2 (BMP2) levels. To investigate the functional consequence of chondrocyte-specific BMP2 expression, we made a collagen type II dependent, doxycycline (dox)-inducible BMP2 transgenic mouse

  12. Collagen-induced expression of collagenase-3 by primary chondrocytes is mediated by integrin alpha 1 and discoidin domain receptor 2 : a protein kinase C-dependent pathway

    NARCIS (Netherlands)

    Vonk, Lucienne A.; Doulabi, Behrouz Z.; Huang, ChunLing; Helder, Marco N.; Everts, Vincent; Bank, Ruud A.

    2011-01-01

    Methods. Goat articular chondrocytes and chondrons were cultured on collagen coatings. Small interfering RNA (siRNA) oligonucleotides targeted against ITG alpha 1 and DDR2 were transfected into primary chondrocytes. Chemical inhibitors for mitogen-activated protein kinase kinase (MEK1) (PD98059), fo

  13. [Autologous fat grafting in children].

    Science.gov (United States)

    Baptista, C; Bertrand, B; Philandrianos, C; Degardin, N; Casanova, D

    2016-10-01

    Lipofilling or fat grafting transfer is defined as a technique of filling soft tissue by autologous fat grafting. The basic principle of lipofilling is based on a harvest of adipose tissue, followed by a reinjection after treatment. Lipofilling main objective is a volume defect filling, but also improving cutaneous trophicity. Lipofilling specificities among children is mainly based on these indications. Complications of autologous fat grafting among children are the same as those in adults: we distinguish short-term complications (intraoperative and perioperative) and the medium and long-term complications. The harvesting of fat tissue is the main limiting factor of the technique, due to low percentage of body fat of children. Indications of lipofilling among children may be specific or similar to those in adults. There are two types of indications: cosmetic, in which the aim of lipofilling is correcting a defect density, acquired (iatrogenic, post-traumatic scar) or malformation (otomandibular dysplasia, craniosynostosis, Parry Romberg syndrom, Poland syndrom, pectus excavatum…). The aim of functional indications is correcting a velar insufficiency or lagophthalmos. In the paediatric sector, lipofilling has become an alternative to the conventional techniques, by its reliability, safety, reproducibility, and good results.

  14. Philadelphia's Renaissance Schools Initiative after Four Years

    Science.gov (United States)

    Stratos, Kati; Wolford, Tonya; Reitano, Adrienne

    2015-01-01

    In 2010-2011, the School District of Philadelphia (the District) launched its Renaissance Schools Initiative, a program designed to dramatically improve student achievement in the District's lowest performing schools. Some schools became Promise Academies, based on the federal turnaround model, and remained District-operated neighborhood schools.…

  15. Neonatal Intestinal Obstruction-Four Year Experience

    Directory of Open Access Journals (Sweden)

    D. Rathore

    2015-06-01

    Full Text Available Aim of study: To study the aetiology and frequency, sex incidence, age of presentation, management and outcome of neonatal intestinal obstruction. Material and Methods: This prospective study of 316 neonates with intestinal obstruction was conducted over a period of 4 years from November 2009 to October 2013 at single institute. These cases were managed by various surgical procedures. Their epidemiology, day of presentation, associated anomalies and outcomes were studied. Results: A total of 316 neonates (277 males and 39 females were operated for intestinal obstruction. 268(84.81% neonates presented in the 1st week of life. Imperforate anus occurred in 206 (65.19%.Small bowel atresia accounted for23 (7.27% cases while duodenal atresia was seen in19 (6.01% patients. Infantile hypertrophic pyloric stenosis and Malrotation each occurred in 14 (4.43% patients; Hirschsprung’s disease in 18(5.69%, Necrotising Enterocolitis in 12(3.79%, Meconium disease of newborn in 9(2.85% while colonic atresia was seen in one (0.3% patient. Colostomy was performed in 145(45.88%, Pouchostomy in 15(4.74% and Cutback anoplasty in 56(17.72% patients. Ramsted’s Pyloromyotomy in 13(4.11%% neonates, Laparoscopic Pyloromyotomy in 1(0.3%,Kimura’s Duodenoduodenostomy in 19(6.01% ,End to Back anastomosis in 24(7.59% , End to End anastomosis in 7(2.21% , Multiple anastomosis in 2(0.6% , Enterotomy with irrigation in 7(2.21% , Ladd’s procedure in 14(4.43% , ,Single stage transanal pull through in 8(2.53% , Ileostomy in 2(0.6% , Single stage Abdominoperineal pull through in 2(0.6%, Levelling colostomy in 6(1.89% ,Peritoneal drain insertion under Local anaesthesia in 5(1.58% . Overall mortality was 13.60%. Conclusion: Intestinal Obstruction is the most common surgical emergency in neonatal period. Early and accurate diagnosis is paramount for proper patient management. The etiology, mode of presentation, morbidity and outcome of surgery of intestinal obstruction in neonates in our study were not significantly different from other studies.

  16. Four years of membership / Toomas Hendrik Ilves

    Index Scriptorium Estoniae

    Ilves, Toomas Hendrik, 1953-

    2008-01-01

    Tagasivaade Eesti nelja-aastasele liikmelisusele Euroopa Liidus ja NATO-s. Juuresoleval fotol Eesti president Toomas Hendrik Ilves ja Läti president Valdis Zatlers Valgas/Valkas enne Eesti ja Läti vahelise piiritõkke eemaldamist seoses Schengeni õigusruumi laienemisega, 21. detsember 2007

  17. Multicystic Dysplastic Kidney: Four-Year Evaluation

    Directory of Open Access Journals (Sweden)

    Berna AYTAÇ

    2011-09-01

    Full Text Available Objective: Multicystic dysplastic kidney is a relatively common developmental abnormality in infants and children. Additional abnormalities like vesicoureteral reflux, ureteropelvic junction obstruction, and ureterovesical junction obstruction may accompany multicystic dysplastic kidney. In this article, we reviewed the pathogenesis, diagnosis, associated urinary tract anomalies and results of management of multicystic dysplastic kidney in the light of the literature.Material and Method: We retrospectively assessed 20 children with unilateral multicystic dysplastic kidney between January 2005 and December 2009. Mean duration of follow-up was 35 ± 8.7 months. All children with multicystic dysplastic kidney underwent nephrectomy. Patient characteristics, the pathology findings, associated urinary tract anomalies and results of management were reviewed.Results: Abdominal ultrasound, voiding cystourethrography and renal scintigraphy revealed vesicoureteral reflux in 3 (15% children, ureteropelvic junction obstruction in 3 (15% children, and a duplex system in the contralateral kidney in 1 (5% child. Blood pressure values, serum urea, creatinine and urinalysis were within normal range in all children during follow-up.Conclusion: A conservative approach to children with multicystic dysplastic kidney has been advocated, but surgical removal is also another management modality on the basis of risk of hypertension, mass effect, potential for malignant change, and cost of repeated ultrasound examination.

  18. The state repatriation programme: four years later

    Directory of Open Access Journals (Sweden)

    Kirillova Yelena

    2010-12-01

    Full Text Available This article analyses the problems of the state voluntary repatriation programme and describes its key functions and implementation mechanisms. The author identifies the causes of deceleration in the repatriation process as well as the weak points of the Programme and the ways to improve it. The article offers data on the resettlement of compatriots over the three years of the Programme implementation, its demographic structure, and the regions of resettlement.

  19. Teaching Styles in Four-Year-Olds.

    Science.gov (United States)

    Feshbach, Norma D.; Devor, Geraldine

    1969-01-01

    This study investigates the relationship between social-class factors and patterns of reinforcement used by preschool children when instructing younger peers. It was hypothesized that middle class Caucasian children would spontaneously use more positive reinforcements and lower class children more negative reinforcements when interacting with…

  20. Evaluating Four Years of ID Experience.

    Science.gov (United States)

    Miles, Gearold D.

    1983-01-01

    Describes the introduction of professionally qualified instructional designers into the Professional Education Division of Arthur Anderson and Co., an international public accounting and professional services firm. The difficulties and misunderstandings that were overcome and the role the instructional designers now play in providing training…

  1. Adolescent Literature Revisited after Four Years.

    Science.gov (United States)

    Donelson, Ken, Ed.

    1976-01-01

    The articles collected in this bulletin survey a wide range of topics concerning adolescent literature. Some of the titles included are "Seven Myths about Adolescent Literature" (Paul B. Janeczko), "The Book as Enemy" (Thomas Weaver), "Popular Non-Fiction Titles for Adolescents" (Noma Russell), "Values and Paperback Power" (Nel Ward), "An Approach…

  2. Viability of porcine nasal septal cartilage grafts following Nd:YAG (n=1.32μm) laser radiation

    Science.gov (United States)

    Chao, Kenneth K. H.; Wong, Brian J.; Kim, Hong K.; Milner, Thomas E.; Sung, Chung-Ho; Sobol, Emil N.; Nelson, J. Stuart

    2000-06-01

    Mechanically deformed morphologic cartilage grafts undergo a temperature dependent phase transformation during sustained laser irradiation that results in reshaping of the specimen. While thermal, optical, and mechanical properties of cartilage undergoing laser heating have been previously investigated, the viability of these irradiated grafts has yet to be examined closely until now. In this study, chondrocyte viability following laser irradiation was determined by measuring the incorporation of radiolabelled sulfate (Na 35SO4-2) into proteoglycan (PTG) macromolecules. Proteoglycans are highly sulfated and are the principal molecular constituents of cartilage matrix. Their synthesis directly reflects chondrocyte viability. By measuring the scintillation counts of 35SO4-2 uptake and normalizing the value by the total protein content of each specimen we can determine the level of PTG synthesis rates following laser reshaping. Regional baseline PTG synthesis rates as a function of location was determined by dividing each specimen into six regions. All regions except the most cephalic are demonstrated similar PTG synthesis rates. The most cephalic region exhibited a significantly greater PTG synthesis rates. In order to establish a positive control for this study, specimens were immersed in boiling saline water for approximately 40 minutes. The boiled specimens demonstrated a fivefold increase in normalized radioisotope uptake and suggest that the non-specific uptake of radioactive Na35SO4-2 is caused by structural alterations in the collagen matrix caused by extensive thermal exposure. To avoid this thermal artifact, another positive control was established using nitric oxide was to induce apoptosis of the chondrocytes, resulting in significantly lower PTG synthesis compared to untreated tissue. Cartilage specimens (25 X 10 X 2 mm) were irradiated with light emitted from an Nd:YAG laser (25 W/cm2, (lambda) equals 1.32 micrometer) while radiometric surface temperature

  3. In vitro cell quality of articular chondrocytes assigned for autologous implantation in dependence of specific patient characteristics

    DEFF Research Database (Denmark)

    Pestka, Jan M; Schmal, Hagen; Salzmann, Gian;

    2011-01-01

    between chondrocyte quality after in vitro cultivation and possible correlations with patient-specific factors. DESIGN: Cell quality of 252 consecutive ACI patients was assessed after chondrocyte in vitro expansion by determination of the expression of cartilage relevant surface marker CD44 and cartilage......-specific differentiation markers (aggrecan and collagen type II). All cell quality parameters were correlated with patient-specific parameters, such as age, size and defect location, number of defects and grade of joint degeneration according to the Kellgren-Lawrence classification. RESULTS: Neither the expression of CD44......, aggrecan or collagen type II nor cell density or viability after proliferation seemed to correlate with the grade of joint degeneration, defect aetiology or patient gender. However, chondrocytes harvested from the knee joints of patients at less than 20 years of age showed significantly higher expression...

  4. Sesamin inhibits IL-1β-stimulated inflammatory response in human osteoarthritis chondrocytes by activating Nrf2 signaling pathway.

    Science.gov (United States)

    Kong, Pengyu; Chen, Guanghua; Jiang, Anlong; Wang, Yufu; Song, Chengchao; Zhuang, Jinpeng; Xi, Chunyang; Wang, Guangxi; Ji, Ye; Yan, Jinglong

    2016-12-13

    Sesamin, a bioactive component extracted from sesame, has been reported to exert anti-inflammatory and anti-oxidant effects. In this study, we evaluated the anti-inflammatory effects of sesamin on IL-1β-stimulated human osteoarthritis chondrocytes and investigated the possible mechanism. Results demonstrated that sesamin treatment significantly inhibited PGE2 and NO production induced by IL-1β. Sesamin inhibited MMP1, MMP3, and MMP13 production in IL-1β-stimulated chondrocytes. Sesamin also inhibited IL-1β-induced phosphorylation of NF-κB p65 and IκBα. Meanwhile, sesamin was found to up-regulate the expression of Nrf2 and HO-1. However, Nrf2 siRNA reversed the anti-inflammatory effects of sesamin. In conclusion, our results suggested that sesamin showed anti-inflammatory effects in IL-1β-stimulated chondrocytes by activating Nrf2 signaling pathway.

  5. Upregulation of aggrecan and type II collagen mRNA expression in bovine chondrocytes by the application of hydrostatic pressure.

    Science.gov (United States)

    Toyoda, Takashi; Seedhom, Bahaa B; Kirkham, Jennifer; Bonass, William A

    2003-01-01

    The aim of this study was to investigate the effect of hydrostatic pressure on the expression of messenger ribonucleic acid (mRNA) for specific extracellular matrix proteins in chondrocytes. Chondrocytes obtained from bovine metatarsophalangeal joints were embedded in cylindrical 2% agarose gels. A novel experimental system was used to apply 5 MPa of static hydrostatic pressure to these chondrocytes for 4 hours. The application of hydrostatic pressure caused a significant increase in the level of aggrecan mRNA by almost four fold (papplication of hydrostatic pressure, in the absence of cell deformation, can bring about changes in the matrix components which may play an important role in the homeostasis and mechanical properties of articular cartilage.

  6. Hybrid Graft Anterior Cruciate Ligament Reconstruction: A Predictable Graft for Knee Stabilization.

    Science.gov (United States)

    Alvarez-Pinzon, Andres M; Barksdale, Leticia; Krill, Michael K; Leo, Brian M

    2015-06-01

    Trauma to the anterior cruciate ligament (ACL) is a season-ending injury and involves months of activity modification and rehabilitation. The annual incidence of ACL tears in the United States is approximately 200,000, which allows for a broad range of individualized treatment options. Various surgical techniques, including transtibial and independent tunnel drilling, allograft and autograft tissue, and various implants, have been described in the literature. This article describes the indications and technique for a hybrid soft tissue graft for ACL reconstruction. Autologous grafts eliminate the risk of disease transmission and have recently been shown to have a lower rerupture rate, particularly in younger, active patients; however, the harvesting of autologous hamstring grafts carries a risk of donor-site morbidity, iatrogenic injury of the graft, and inadequate graft size. In contrast to a traditional autologous soft tissue graft, the hybrid graft allows for graft size customization for a desired reconstruction, especially in cases where autograft hamstrings may be iatrogenically damaged or of inadequate size when harvested. The goal of a hybrid graft ACL reconstruction is to provide a favorable-sized graft with clinical outcomes comparable with autologous soft tissue grafts. In contrast to a traditional autologous soft tissue graft, this technique provides another option in the event of unforeseen deficiencies or complications associated with harvesting and preparation of the autologous gracilis and semitendinosis soft tissue graft.

  7. Evaluating Joint Morbidity after Chondral Harvest for Autologous Chondrocyte Implantation (ACI)

    Science.gov (United States)

    McCarthy, Helen S.; Richardson, James B.; Parker, Jane C. E.; Roberts, Sally

    2016-01-01

    Objective To establish if harvesting cartilage to source chondrocytes for autologous chondrocyte implantation (ACI) results in donor site morbidity. Design Twenty-three patients underwent ACI for chondral defects of either the ankle or the hip. This involved cartilage harvest from the knee (stage I), chondrocyte expansion in the laboratory and implantation surgery (stage II) into the affected joint. Prior to chondral harvest, no patient had sought treatment for their knee. Lysholm knee scores were completed prior to chondral harvest and annually post-ACI. Histological analyses of the donor site were performed at 12.3 ± 1.5 months for 3 additional patients who had previously had ACI of the knee. Results The median preoperative Lysholm score was 100, with no significant differences observed at either 13.7±1.7 months or 4.8±1.8 years postharvest (median Lysholm scores 91.7 and 87.5, respectively). Patients whose cartilage was harvested from the central or medial trochlea had a significantly higher median Lysholm score at latest follow-up (97.9 and 93.4, respectively), compared with those taken from the intercondylar notch (median Lysholm score 66.7). The mean International Cartilage Repair Society (ICRS) II histological score for the biopsies taken from the donor site of 3 additional knee ACI patients was 117 ± 10 (maximum score 140). Conclusions This study suggests that the chondral harvest site in ACI is not associated with significant joint morbidity, at least up to 5 years postharvest. However, one should carefully consider the location for chondral harvest as this has been shown to affect knee function in the longer term. PMID:26958313

  8. The signs of differentiation of chondrocytes in the formation of early cartilage lesions in the elderly

    Directory of Open Access Journals (Sweden)

    Elena Vasil'evna Chetina

    2011-01-01

    Results. The activity of collagen type II cleavage was shown to be increased in the area of age-related OA-like cartilage lesions. This was accompanied by the high expression of collagenases of metalloproteinases (MMP 1, 14 (MT1-MMP, aggrecanases - desintegrin and MMP with thrombospondin type 1 motif (ADAMTS 5, the cytokines of interleukins (IL 1α/β and tumor necrosis factor-α (TNF-α, as well as the genes associated with chondrocyte hypertrophy of type X collagen (C0L10A1, MMP 13 and 9, Indian hedgehog (Ihh and cas-pase 3 in the immediate vicinity of a lesion area. At the same time, there was a high expression of growth factors associated with the proliferation phase of chondrocytes, namely: parathyroid hormone-related peptide (PTHrP, fibroblast growth factor-2 (FGF-2, transforming growth factor β1/2 (TGF-β1/2, as well as macromolecules of matrix of type II collagen (C0L2A1 and aggrecan in both the areas adjacent to the lesions and at a considerable distance from their center. However, these areas showed no higher collagen cleavage activity. Nether higher collagen cleavage, nor excess expression of the genes examined were observed in the absolutely intact cartilage areas. Conclusion. Our studies have indicated that the area of very early age-related OA-like focal cartilage lesions exhibits enhanced type II collagen cleavage that is attended by the expression of the genes associated with chondrocyte differentiation in the embryonic growth plate.

  9. Subculture of chondrocytes on a collagen type I-coated substrate with suppressed cellular dedifferentiation.

    Science.gov (United States)

    Kino-Oka, Masahiro; Yashiki, Shino; Ota, Yuka; Mushiaki, Yuko; Sugawara, Katsura; Yamamoto, Takeyuki; Takezawa, Toshiaki; Taya, Masahito

    2005-01-01

    To evaluate the degree of cellular dedifferentiation, subculture of chondrocytes was conducted on a surface coated with collagen type I at a density of 1.05 mg/cm(2). In the primary culture, most of the cells were round in shape on the collagen (CL) substrate, whereas fibroblastic and partially extended cells were dominant on the polystyrene plastic (PS) substrate. Stereoscopic observation revealed that the round-shaped cells on the CL substrate were hemispherical with nebulous and punctuated F-actin filaments, whereas the fibroblastic cells on the PS substrate were flattened with fully developed stress fibers. This suggested that cell polarization was suppressed during culture on the former substrate. Although serial passages of chondrocytes through subcultures on the CL and PS substrates caused a decrease in the number of round-shaped cells, the morphological change was appreciably suppressed on the CL substrate, as compared with that on the PS substrate. It was found that only round-shaped cells formed collagen type II, which supports the view that cellular dedifferentiation can be suppressed to some extent on the CL substrate. Three-dimensional cultures in collagen gel were performed with cells isolated freshly and passaged on the CL or PS substrate. Cell density at 21 days in the culture of cells passaged on the CL substrate was comparable to that in the culture of freshly isolated cells, in spite of a significant reduction in cell density observed in the culture of cells passaged on the PS substrate. In addition, histological analysis revealed that the expression of glycosaminoglycans and collagen type II was of significance in the collagen gel with cells passaged on the CL substrate, and likewise in the gel with freshly isolated cells. This indicated that the CL substrate could offer a monolayer culture system for expanding chondrocyte cells.

  10. Losartan increases bone mass and accelerates chondrocyte hypertrophy in developing skeleton.

    Science.gov (United States)

    Chen, Shan; Grover, Monica; Sibai, Tarek; Black, Jennifer; Rianon, Nahid; Rajagopal, Abbhirami; Munivez, Elda; Bertin, Terry; Dawson, Brian; Chen, Yuqing; Jiang, Ming-Ming; Lee, Brendan; Yang, Tao; Bae, Yangjin

    2015-05-01

    Angiotensin receptor blockers (ARBs) are a group of anti-hypertensive drugs that are widely used to treat pediatric hypertension. Recent application of ARBs to treat diseases such as Marfan syndrome or Alport syndrome has shown positive outcomes in animal and human studies, suggesting a broader therapeutic potential for this class of drugs. Multiple studies have reported a benefit of ARBs on adult bone homeostasis; however, its effect on the growing skeleton in children is unknown. We investigated the effect of Losartan, an ARB, in regulating bone mass and cartilage during development in mice. Wild type mice were treated with Losartan from birth until 6 weeks of age, after which bones were collected for microCT and histomorphometric analyses. Losartan increased trabecular bone volume vs. tissue volume (a 98% increase) and cortical thickness (a 9% increase) in 6-weeks old wild type mice. The bone changes were attributed to decreased osteoclastogenesis as demonstrated by reduced osteoclast number per bone surface in vivo and suppressed osteoclast differentiation in vitro. At the molecular level, Angiotensin II-induced ERK1/2 phosphorylation in RAW cells was attenuated by Losartan. Similarly, RANKL-induced ERK1/2 phosphorylation was suppressed by Losartan, suggesting a convergence of RANKL and angiotensin signaling at the level of ERK1/2 regulation. To assess the effect of Losartan on cartilage development, we examined the cartilage phenotype of wild type mice treated with Losartan in utero from conception to 1 day of age. Growth plates of these mice showed an elongated hypertrophic chondrocyte zone and increased Col10a1 expression level, with minimal changes in chondrocyte proliferation. Altogether, inhibition of the angiotensin pathway by Losartan increases bone mass and accelerates chondrocyte hypertrophy in growth plate during skeletal development.

  11. Characterization of human primary chondrocytes of osteoarthritic cartilage at varying severity

    Institute of Scientific and Technical Information of China (English)

    YIN Jing; YANG Zheng; CAO Yong-ping; GE Zi-gang

    2011-01-01

    Background There is a difficulty in evaluating the in vivo functionality of individual chondrocytes,and there is much heterogeneity among cartilage affected by osteoarthritis (OA).In this study,in vitro cultured chondrocytes harvested from varying stages of degeneration were studied as a projective model to further understand the pathogenesis of osteoarthritis.Methods Cartilage of varying degeneration of end-stage OA was harvested,while cell yield and matrix glycosaminoglycan (GAG) content were measured.Cell morphology,proliferation,and gene expression of collagen type Ⅰ,Ⅱ,and Ⅹ,aggrecan,matrix metalloproteinase 13 (MMP-13),and ADAMTS5 of the acquired chondrocytes were measured during subsequent in vitro culture.Results Both the number of cells and the GAG content increased with increasing severity of OA.Cell spreading area increased and gradually showed spindle-like morphology during in vitro culture.Gene expression of collagen type Ⅱ,collagen type X as well as GAG decreased with severity of cartilage degeneration,while expression of collagen type Ⅰ increased.Expression of MMP-13 increased with severity of cartilage degeneration,while expression of ADAMTS-5 remained stable.Expression of collagen type Ⅱ,X,GAG,and MMP-13 substantially decreased with in vitro culture.Expression of collagen type Ⅰ increased with in vitro cultures,while expression of ADAMTS 5 remained stable.Conclusions Expression of functional genes such as collagen type Ⅱ and GAG decreased during severe degeneration of OA cartilage and in vitro dedifferentiation.Gene expression of collagen Ⅰ and MMP-13 increased with severity of cartilage degeneration.

  12. Human-like collagen/nano-hydroxyapatite scaffolds for the culture of chondrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Jia, Liping; Duan, Zhiguang [Shaanxi Key Laboratory of Degradable Biomedical Materials, Northwest University, 229 Taibai North Road, Xi' an, Shaanxi 710069 (China); Shaanxi R and D Center of Biomaterials and Fermentation Engineering, Northwest University, 229 Taibai North Road, Xi' an, Shaanxi 710069 (China); Fan, Daidi, E-mail: fandaidi@nwu.edu.cn [Shaanxi Key Laboratory of Degradable Biomedical Materials, Northwest University, 229 Taibai North Road, Xi' an, Shaanxi 710069 (China); Shaanxi R and D Center of Biomaterials and Fermentation Engineering, Northwest University, 229 Taibai North Road, Xi' an, Shaanxi 710069 (China); Mi, Yu; Hui, Junfeng [Shaanxi Key Laboratory of Degradable Biomedical Materials, Northwest University, 229 Taibai North Road, Xi' an, Shaanxi 710069 (China); Shaanxi R and D Center of Biomaterials and Fermentation Engineering, Northwest University, 229 Taibai North Road, Xi' an, Shaanxi 710069 (China); Chang, Le [School of Chemical Engineering, Northwest University, Xi' an, Shaanxi 710069 (China)

    2013-03-01

    Three dimensional (3D) biodegradable porous scaffolds play a key role in cartilage tissue repair. Freeze-drying and cross-linking techniques were used to fabricate a 3D composite scaffold that combined the excellent biological characteristics of human-like collagen (HLC) and the outstanding mechanical properties of nano-hydroxyapatite (nHA). The scaffolds were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and compression tests, using Relive Registered-Sign Artificial Bone (RAB) scaffolds as a control. HLC/nHA scaffolds displayed homogeneous interconnected macroporous structure and could withstand a compression stress of 2.67 {+-} 0.37 MPa, which was higher than that of the control group. Rabbit chondrocytes were seeded on the composite porous scaffolds and cultured for 21 days. Cell/scaffold constructs were examined using SEM, histological procedures, and biochemical assays for cell proliferation and the production of glycosaminoglycans (GAGs). The results indicated that HLC/nHA porous scaffolds were capable of encouraging cell adhesion, homogeneous distribution and abundant GAG synthesis, and maintaining natural chondrocyte morphology compared to RAB scaffolds. In conclusion, the presented data warrants the further exploration of HLC/nHA scaffolds as a potential biomimetic platform for chondrocytes in cartilage tissue engineering. - Highlights: Black-Right-Pointing-Pointer Human-like collagen was first used to prepare cartilage tissue engineering scaffold. Black-Right-Pointing-Pointer Genipin, a natural biological cross-linking agent, was introduced to treat scaffold. Black-Right-Pointing-Pointer We chose market product as a control.

  13. In vitro human chondrocyte culture on plasma-treated poly(glycerol sebacate) scaffolds.

    Science.gov (United States)

    Theerathanagorn, Tharinee; Klangjorhor, Jeerawan; Sakulsombat, Morakot; Pothacharoen, Peraphan; Pruksakorn, Dumnoensun; Kongtawelert, Prachya; Janvikul, Wanida

    2015-01-01

    Porous poly(glycerol sebacate) (PGS) scaffolds were prepared using a salt leaching technique and subsequently surface modified by a low oxygen plasma treatment prior to the use in the in vitro culture of human chondrocytes. Condensation polymerization of glycerol and sebacic acid used at various mole ratios, i.e. 1:1, 1:1.25, and 1:1.5, was initially conducted to prepare PGS prepolymers. Porous elastomeric PGS scaffolds were directly fabricated from the mixtures of each prepolymer and 90% (w/w) NaCl particles and then subjected to the plasma treatment to enhance the surface hydrophilicity of the materials. The properties of both untreated and plasma-treated PGS scaffolds were comparatively evaluated, in terms of surface morphology, surface chemical composition, porosity, and storage modulus using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy, micro-computed tomography, and dynamic mechanical analysis, respectively. The responses of chondrocytes cultured on individual PGS scaffolds were assessed, in terms of cell proliferation and ECM production. The results revealed that average pore sizes and porosity of the scaffolds were increased with an increasing sebacic acid concentration used. The storage moduli of the scaffolds were raised after the plasma treatment, possibly due to the further crosslinking of PGS upon treatment. Moreover, the scaffold prepared with a higher sebacic acid content demonstrated a greater capability of promoting cell infiltration, proliferation, and ECM production, especially when it was plasma-treated; the greatest HA, sGAG, uronic acid, and collagen contents were detected in matrix of this scaffold. The H & E and safranin O staining results also strongly supported this finding. The storage modulus of the scaffold was intensified after incubation with the chondrocytes for 21 days, indicating the accretion and retention of matrix ECM on the cell-cultured scaffold.

  14. New insight on FGFR3-related chondrodysplasias molecular physiopathology revealed by human chondrocyte gene expression profiling.

    Directory of Open Access Journals (Sweden)

    Laurent Schibler

    Full Text Available Endochondral ossification is the process by which the appendicular skeleton, facial bones, vertebrae and medial clavicles are formed and relies on the tight control of chondrocyte maturation. Fibroblast growth factor receptor (FGFR3 plays a role in bone development and maintenance and belongs to a family of proteins which differ in their ligand affinities and tissue distribution. Activating mutations of the FGFR3 gene lead to craniosynostosis and multiple types of skeletal dysplasia with varying degrees of severity: thanatophoric dysplasia (TD, achondroplasia and hypochondroplasia. Despite progress in the characterization of FGFR3-mediated regulation of cartilage development, many aspects remain unclear. The aim and the novelty of our study was to examine whole gene expression differences occurring in primary human chondrocytes isolated from normal cartilage or pathological cartilage from TD-affected fetuses, using Affymetrix technology. The phenotype of the primary cells was confirmed by the high expression of chondrocytic markers. Altered expression of genes associated with many cellular processes was observed, including cell growth and proliferation, cell cycle, cell adhesion, cell motility, metabolic pathways, signal transduction, cell cycle process and cell signaling. Most of the cell cycle process genes were down-regulated and consisted of genes involved in cell cycle progression, DNA biosynthesis, spindle dynamics and cytokinesis. About eight percent of all modulated genes were found to impact extracellular matrix (ECM structure and turnover, especially glycosaminoglycan (GAG and proteoglycan biosynthesis and sulfation. Altogether, the gene expression analyses provide new insight into the consequences of FGFR3 mutations in cell cycle regulation, onset of pre-hypertrophic differentiation and concomitant metabolism changes. Moreover, impaired motility and ECM properties may also provide clues about growth plate disorganization. These

  15. An evaluation of chondrocyte morphology and gene expression on superhydrophilic vertically-aligned multi-walled carbon nanotube films

    Energy Technology Data Exchange (ETDEWEB)

    Antonioli, Eliane, E-mail: eliane.antonioli@einstein.br [Research and Education Institute, Hospital Israelita Albert Einstein, Sao Paulo, SP (Brazil); Lobo, Anderson O., E-mail: aolobo@univap.br [Laboratory of Biomedical Nanotechnology, Universidade do Vale do Paraiba, Sao Jose dos Campos, Sao Paulo (Brazil); Ferretti, Mario, E-mail: ferretti@einstein.br [Research and Education Institute, Hospital Israelita Albert Einstein, Sao Paulo, SP (Brazil); Ortophedic Division, Federal University of Sao Paulo, SP (Brazil); Cohen, Moises, E-mail: m.cohen@uol.com.br [Research and Education Institute, Hospital Israelita Albert Einstein, Sao Paulo, SP (Brazil); Ortophedic Division, Federal University of Sao Paulo, SP (Brazil); Marciano, Fernanda R., E-mail: femarciano@uol.com.br [Laboratory of Biomedical Nanotechnology, Universidade do Vale do Paraiba, Sao Jose dos Campos, Sao Paulo (Brazil); Corat, Evaldo J., E-mail: corat@las.inpe.br [Laboratorio Associado de Sensores e Materiais, Instituto Nacional de Pesquisas Espaciais, Sao Jose dos Campos, Sao Paulo (Brazil); Trava-Airoldi, Vladimir J., E-mail: vladimir@las.inpe.br [Laboratorio Associado de Sensores e Materiais, Instituto Nacional de Pesquisas Espaciais, Sao Jose dos Campos, Sao Paulo (Brazil)

    2013-03-01

    Cartilage serves as a low-friction and wear-resistant articulating surface in diarthrodial joints and is also important during early stages of bone remodeling. Recently, regenerative cartilage research has focused on combinations of cells paired with scaffolds. Superhydrophilic vertically aligned carbon nanotubes (VACNTs) are of particular interest in regenerative medicine. The aim of this study is to evaluate cell expansion of human articular chondrocytes on superhydrophilic VACNTs, as well as their morphology and gene expression. VACNT films were produced using a microwave plasma chamber on Ti substrates and submitted to an O{sub 2} plasma treatment to make them superhydrophilic. Human chondrocytes were cultivated on superhydrophilic VACNTs up to five days. Quantitative RT-PCR was performed to measure type I and type II Collagen, Sox9, and Aggrecan mRNA expression levels. The morphology was analyzed by scanning electron microscopy (SEM) and confocal microscopy. SEM images demonstrated that superhydrophilic VACNTs permit cell growth and adhesion of human chondrocytes. The chondrocytes had an elongated morphology with some prolongations. Chondrocytes cultivated on superhydrophilic VACNTs maintain the level expression of Aggrecan, Sox9, and Collagen II determined by qPCR. This study was the first to indicate that superhydrophilic VACNTs may be used as an efficient scaffold for cartilage or bone repair. Highlights: Black-Right-Pointing-Pointer Chondrocytes were cultivated on Superhydrophilic Vertically Aligned Multiwall Carbon Nanotubes (VACNT). Black-Right-Pointing-Pointer We have shown a correlation between gene expression and thermodynamics aspects. Black-Right-Pointing-Pointer Superhydrhophilic VACNT will be an excellent substrate for cartilage and bone tissue regeneration.

  16. Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes

    Directory of Open Access Journals (Sweden)

    M. Lehmann

    2013-11-01

    Full Text Available Cartilage regeneration based on isolated and culture-expanded chondrocytes has been studied in various in vitro models, but the quality varies with respect to the morphology and the physiology of the synthesized tissues. The aim of our study was to promote in vitro chondrogenesis of human articular chondrocytes using a novel three-dimensional (3-D cultivation system in combination with the chondrogenic differentiation factors transforming growth factor beta 2 (TGF-b2 and L-ascorbic acid. Articular chondrocytes isolated from six elderly patients were expanded in monolayer culture. A single-cell suspension of the dedifferentiated chondrocytes was then added to agar-coated dishes without using any scaffold material, in the presence, or absence of TGF-b2 and/or L-ascorbic acid. Three-dimensional cartilage-like constructs, called single spheroids, and microtissues consisting of several spheroids fused together, named as fusions, were formed. Generated tissues were mainly characterized using histological and immunohistochemical techniques. The morphology of the in vitro tissues shared some similarities to native hyaline cartilage in regard to differentiated S100-positive chondrocytes within a cartilaginous matrix, with strong collagen type II expression and increased synthesis of proteoglycans. Finally, our innovative scaffold-free fusion culture technique supported enhanced chondrogenesis of human articular chondrocytes in vitro. These 3-D hyaline cartilage-like microtissues will be useful for in vitro studies of cartilage differentiation and regeneration, enabling optimization of functional tissue engineering and possibly contributing to the development of new approaches to treat traumatic cartilage defects or osteoarthritis.

  17. Mesenchymal stem cells from adipose tissue which have been differentiated into chondrocytes in three-dimensional culture express lubricin.

    Science.gov (United States)

    Musumeci, Giuseppe; Lo Furno, Debora; Loreto, Carla; Giuffrida, Rosario; Caggia, Silvia; Leonardi, Rosalia; Cardile, Venera

    2011-11-01

    The present study focused on the isolation, cultivation and characterization of human mesenchymal stem cells (MSCs) from adipose tissue and on their differentiation into chondrocytes through the NH ChondroDiff medium. The main aim was to investigate some markers of biomechanical quality of cartilage, such as lubricin, and collagen type I and II. Little is known, in fact, about the ability of chondrocytes from human MSCs of adipose tissue to generate lubricin in three-dimensional (3D) culture. Lubricin, a 227.5-kDa mucinous glycoprotein, is known to play an important role in articular joint physiology, and the loss of accumulation of lubricin is thought to play a role in the pathology of osteoarthritis. Adipose tissue is an alternative source for the isolation of multipotent MSCs, which allows them to be obtained by a less invasive method and in larger quantities than from other sources. These cells can be isolated from cosmetic liposuctions in large numbers and easily grown under standard tissue culture conditions. 3D chondrocytes were assessed by histology (hematoxylin and eosin) and histochemistry (Alcian blue and Safranin-O/fast green staining). Collagen type I, II and lubricin expression was determined through immunohistochemistry and Western blot. The results showed that, compared with control cartilage and monolayer chondrocytes showing just collagen type I, chondrocytes from MSCs (CD44-, CD90- and CD105- positive; CD45-, CD14- and CD34-negative) of adipose tissue grown in nodules were able to express lubricin, and collagen type I and II, indicative of hyaline cartilage formation. Based on the function of lubricin in the joint cavity and disease and as a potential therapeutic agent, our results suggest that MSCs from adipose tissue are a promising cell source for tissue engineering of cartilage. Our results suggest that chondrocyte nodules producing lubricin could be a novel biotherapeutic approach for the treatment of cartilage abnormalities.

  18. A novel rat tail collagen type-I gel for the cultivation of human articular chondrocytes in low cell density.

    Science.gov (United States)

    Muller-Rath, R; Gavénis, K; Andereya, S; Mumme, T; Schmidt-Rohlfing, B; Schneider, U

    2007-12-01

    Collagen type-I matrix systems have gained growing importance as a cartilage repair device. However, most of the established matrix systems use collagen type-I of bovine origin seeded in high cell densities. Here we present a novel collagen type-I gel system made of rat tail collagen for the cultivation of human chondrocytes in low cell densities. Rat tail collagen type-I gel (CaReS, Arthro Kinetics, Esslingen, Germany) was seeded with human passage 2 chondrocytes in different cell densities to evaluate the optimal cell number. In vitro, the proliferation factor of low density cultures was more than threefold higher compared with high density cultures. After 6 weeks of in vitro cultivation, freshly prepared chondrocytes with an initial cell density of 2x10(5) cells/mL showed a proliferation factor of 33. A cell density of 2x10(5) cells/mL was chosen for in vitro and in vivo cultivation using the common nude mouse model as an in vivo system. Chondrocytes stayed viable as a Live/Dead fluorescence assay and TUNEL staining revealed. During in vitro cultivation, passage 0 cells partly dedifferentiated morphologically. In vivo, passage 0 cells maintained the chondrocyte phenotype and demonstrated an increased synthesis of collagen type-II protein and gene expression compared to passage 2 cells. Passage 2 cells did not redifferentiate in vivo. Cultivating a cell-seeded collagen gel of bovine origin as a control (AtelocollagenTM, Koken, Tokyo, Japan) did not lead to superior results with regard to cell morphology, col-II protein production and col-II gene expression. With the CaReS collagen gel system the best quality of repair tissue was obtained by seeding freshly isolated chondrocytes.

  19. MicroRNA-146a Induced by Hypoxia Promotes Chondrocyte Autophagy through Bcl-2

    Directory of Open Access Journals (Sweden)

    Fei Zhang

    2015-10-01

    Full Text Available Background/Aims: There have been many studies on the etiology of osteoarthritis (OA with regard to the function of inflammatory cytokines, the process of cartilage degradation, the function of miR-146a, hypoxia stimulation and autophagy in OA chondrocytes, but there have been no reports on the relationship between miR-146a and autophagy in cartilage, especially under hypoxia. This study aimed to confirm the relationship of miR-146a and autophagy in cartilage under hypoxia. Methods: Chondrocytes were treated by hypoxia gradients, and the main factors including HIF-1α, HIF-2α, miR-146a and Bcl-2 and autophagy markers ULK-1, ATG-5 were detected by quantitative PCR (Q-PCR and western blotting. The autophagy marker LC-3 was detected by immunofluorescence. The reciprocal effects between miR-146a and Bcl-2 were confirmed by several combinations of shRNAs and adenovirus-gene systems followed by Q-PCR and western blot detection. Results: Hypoxia maintained the chondrocytes phenotype and promoted autophagy and miR-146a expression via HIF-1α, but not HIF-2α, while miR-146a did not reversely affect HIF-1α. The autophagy induced by hypoxia through HIF-1α, miR-146a and Bcl-2. Simply, hypoxia induced HIF-1α, and HIF-1α increased miR-146a, but miR-146a suppressed Bcl-2, an autophagy inhibitor. While Bcl-2 affected neither HIF-1α nor miR-146a. The absence of both HIF-1α and miR-146a or Bcl-2 over-expression inhibited hypoxia-induced autophagy. Conclusion: HIF-1α, miR-146a and Bcl-2 play crucial roles during hypoxia-induced autophagy, Hypoxia, HIF-1α and miR-146a promote chondrocytes autophagy via depressing Bcl-2. We conclude that miR-146a may serve as a novel therapeutic target for protecting cartilage from degeneration in OA.

  20. Dynamic cyclic compression modulates the chondrogenic phenotype in human chondrocytes from late stage osteoarthritis.

    Science.gov (United States)

    Diao, Hua Jia; Fung, Hon Sing; Yeung, Pan; Lam, K L; Yan, Chun Hoi; Chan, Barbara Pui

    2017-02-16

    Human osteoarthritic chondrocytes (hOACs) are characterized by their "dedifferentiated" and catabolic phenotype and lack the ability for restoring their inherent functions by themselves. Here we investigated whether extrinsically supplemented mechanical signal via compression loading would affect the phenotype of hOACs. Specifically, we applied cyclic compression loading on cultured hOACs-collagen constructs and measured the expression of the major chondrogenic factors, cell-matrix interaction molecules and matrix degradation enzymes. Dynamic compression loading stimulates the expression and nuclear localization of sox9 in hOACs and reduces the catabolic events via downregulated expression of collagenases. These results contribute to better understanding towards mechanoregulation of hOACs.

  1. Twist and its effect on ACL graft forces

    NARCIS (Netherlands)

    Arnold, MP; Blankevoort, L; ten Ham, A; Verdonschot, N; van Kampen, A

    2004-01-01

    Graft tension is a controversial topic in anterior cruciate ligament (ACL) surgery. Evidence suggests a narrow range of graft tensions, which allow the graft to remodel to a stable and mature neoligament. In previous cadaver experiments, we showed that twisting the graft could modulate the graft for

  2. Twist and its effect on ACL graft forces.

    NARCIS (Netherlands)

    Arnold, M.P.; Blankevoort, L.; Ham, A. ten; Verdonschot, N.J.J.; Kampen, A. van

    2004-01-01

    Graft tension is a controversial topic in anterior cruciate ligament (ACL) surgery. Evidence suggests a narrow range of graft tensions, which allow the graft to remodel to a stable and mature neoligament. In previous cadaver experiments, we showed that twisting the graft could modulate the graft for

  3. Fixation of tibial plateau fractures with synthetic bone graft versus natural bone graft: a comparison study.

    LENUS (Irish Health Repository)

    Ong, J C Y

    2012-06-01

    The goal of this study was to determine differences in fracture stability and functional outcome between synthetic bone graft and natural bone graft with internal fixation of tibia plateau metaphyseal defects.

  4. Transforming growth factor beta signaling is essential for the autonomous formation of cartilage-like tissue by expanded chondrocytes.

    Directory of Open Access Journals (Sweden)

    Adel Tekari

    Full Text Available Cartilage is a tissue with limited self-healing potential. Hence, cartilage defects require surgical attention to prevent or postpone the development of osteoarthritis. For cell-based cartilage repair strategies, in particular autologous chondrocyte implantation, articular chondrocytes are isolated from cartilage and expanded in vitro to increase the number of cells required for therapy. During expansion, the cells lose the competence to autonomously form a cartilage-like tissue, that is in the absence of exogenously added chondrogenic growth factors, such as TGF-βs. We hypothesized that signaling elicited by autocrine and/or paracrine TGF-β is essential for the formation of cartilage-like tissue and that alterations within the TGF-β signaling pathway during expansion interfere with this process. Primary bovine articular chondrocytes were harvested and expanded in monolayer culture up to passage six and the formation of cartilage tissue was investigated in high density pellet cultures grown for three weeks. Chondrocytes expanded for up to three passages maintained the potential for autonomous cartilage-like tissue formation. After three passages, however, exogenous TGF-β1 was required to induce the formation of cartilage-like tissue. When TGF-β signaling was blocked by inhibiting the TGF-β receptor 1 kinase, the autonomous formation of cartilage-like tissue was abrogated. At the initiation of pellet culture, chondrocytes from passage three and later showed levels of transcripts coding for TGF-β receptors 1 and 2 and TGF-β2 to be three-, five- and five-fold decreased, respectively, as compared to primary chondrocytes. In conclusion, the autonomous formation of cartilage-like tissue by expanded chondrocytes is dependent on signaling induced by autocrine and/or paracrine TGF-β. We propose that a decrease in the expression of the chondrogenic growth factor TGF-β2 and of the TGF-β receptors in expanded chondrocytes accounts for a decrease

  5. CHEMICAL MORPHOLOGY IN GRAFTING ACRYLAMIDE TO POLYETHYLENE

    Institute of Scientific and Technical Information of China (English)

    S. Termnak; K. Sintasanai; T. Amomsakchai; T. Nipithakul; D. Triampo

    2008-01-01

    The scanning force microscopy (SFM)/chemical force microscopy (CFM) were used to study the growth of grafted polyacrylamide (PAM) chains onto polyethylene (PE)-film with varying grafting time. Results from the CFM reveal reduced interaction between the probe and areas with grafted-PAM on the surface. The topography and the friction trace-minus-retrace (TMR) images are complementary to one another resulting from the reduced interaction of the probe that has specificity to chemical domains.

  6. Outcomes of AV Fistulas and AV Grafts after Interventional Stent-Graft Deployment in Haemodialysis Patients

    Energy Technology Data Exchange (ETDEWEB)

    Schmelter, Christopher, E-mail: christopher.schmelter@klinikum-ingolstadt.de; Raab, Udo, E-mail: udo.raab@klinikum-ingolstadt.de [Klinikum Ingolstadt, Department of Diagnostic and Interventional Radiology (Germany); Lazarus, Friedrich, E-mail: friedrich.lazarus@klinikum-ingolstadt.de [Klinikum Ingolstadt, Department of Nephrology (Germany); Ruppert, Volker, E-mail: volker.ruppert@klinikum-ingolstadt.de [Klinikum Ingolstadt, Department of Vascular Surgery (Germany); Vorwerk, Dierk, E-mail: dierk.vorwerk@klinikum-ingolstadt.de [Klinikum Ingolstadt, Department of Diagnostic and Interventional Radiology (Germany)

    2015-08-15

    PurposeThe study was designed to assess outcomes of arteriovenous (AV) accesses after interventional stent-graft deployment in haemodialysis patients.Materials and Methods63 haemodialysis patients with 66 AV fistulas and AV grafts were treated by interventional stent-graft deployment from 2006 to 2012 at our hospital. Data of these patients were retrospectively analysed for location of deployed stent-grafts, occurrence and location of (re-)stenosis and (re-)thrombosis. Complex stenosis was the most frequent indication for stent-graft deployment (45.5 %), followed by complications of angioplasty with vessel rupture or dissection (31.8 %).ResultsA high rate of procedural success was achieved (98.5 %). The most frequent location of the deployed stent-graft was the draining vein (66.7 %). Stent-graft deployment was more frequent in AV grafts than in AV fistulas. Primary patency was 45.5 % at 6 month, 31.3 % at 12 month and 19.2 % at 24 month. Primary patency was significantly better for AV fistulas than for AV grafts with deployed stent-grafts. Patency of the deployed stent-graft was much better than overall AV access primary patency with deployed stent-graft. Re-stenosis with thrombosis was the most frequent indication for re-intervention. Most frequent location of re-stenosis was the draining vein (37.1 %), followed by stenosis at the AV access (29.5 %) and the deployed stent-graft (23.5 %).ConclusionRe-stenosis and re-thrombosis remain frequent in AV fistulas and AV grafts in haemodialysis patients despite stent-graft deployment. Re-stenosis of the deployed stent-graft is, only in the minority of the cases, responsible for AV access dysfunction.

  7. The autologus graft of epithelial tissue culture

    Directory of Open Access Journals (Sweden)

    Minaee B

    1999-08-01

    Full Text Available With the intention of research about culture and autologus graft of epithelial tissue we used 4 french Albino Rabbits with an average age of 2 months. After reproduction on the support in EMEM (Eagle's Minimum Essential Medium we used this for graft after 4 weeks. This region which grafted total replaced. After fixation of this sample and passing them through various process, histological sections were prepared. These sections were stained with H & E and masson's trichrome and studied by light microscope. We succeeded in graft. We hope in the near future by using the method of epithelium tissue culture improving to treat burned patients.

  8. TNF/TNFR{sub 1} pathway and endoplasmic reticulum stress are involved in ofloxacin-induced apoptosis of juvenile canine chondrocytes

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Fu-Tao; Ding, Yi; Shah, Zahir; Xing, Dan; Gao, Yuan; Liu, Dong Ming; Ding, Ming-Xing, E-mail: dmx@mail.hzau.edu.cn

    2014-04-15

    Background and purpose: Quinolones cause obvious cartilaginous lesions in juvenile animals by chondrocyte apoptosis, which results in the restriction of their use in pediatric and adolescent patients. Studies showed that chondrocytes can be induced to produce TNFα, and the cisternae of the endoplasmic reticulum in quinolone-treated chondrocytes become dilated. We investigated whether TNF/TNFR{sub 1} pathway and endoplasmic reticulum stress (ERs) are involved in ofloxacin (a typical quinolone)-induced apoptosis of juvenile canine chondrocytes. Experimental approach: Canine juvenile chondrocytes were treated with ofloxacin. Cell survival and apoptosis rates were determined with MTT method and flow cytometry, respectively. The gene expression levels of the related signaling molecules (TNFα, TNFR{sub 1}, TRADD, FADD and caspase-8) in death receptor pathways and main apoptosis-related molecules (calpain, caspase-12, GADD153 and GRP78) in ERs were measured by qRT-PCR. The gene expression of TNFR{sub 1} was suppressed with its siRNA. The protein levels of TNFα, TNFR{sub 1} and caspase-12 were assayed using Western blotting. Key results: The survival rates decreased while apoptosis rates increased after the chondrocytes were treated with ofloxacin. The mRNA levels of the measured apoptosis-related molecules in death receptor pathways and ERs, and the protein levels of TNFα, TNFR{sub 1} and caspase-12 increased after the chondrocytes were exposed to ofloxacin. The downregulated mRNA expressions of TNFR{sub 1}, Caspase-8 and TRADD, and the decreased apoptosis rates of the ofloxacin-treated chondrocytes occurred after TNFR{sub 1}–siRNA interference. Conclusions and implications: Ofloxacin-induced chondrocyte apoptosis in a time- and concentration-dependent fashion. TNF/TNFR{sub 1} pathway and ERs are involved in ofloxacin-induced apoptosis of juvenile canine chondrocytes in the early stage. - Highlights: • Chondrocyte apoptosis is induced by ofloxacin in a time- and

  9. Regeneration of human-ear-shaped cartilage by co-culturing human microtia chondrocytes with BMSCs.

    Science.gov (United States)

    Zhang, Lu; He, Aijuan; Yin, Zongqi; Yu, Zheyuan; Luo, Xusong; Liu, Wei; Zhang, Wenjie; Cao, Yilin; Liu, Yu; Zhou, Guangdong

    2014-06-01

    Previously, we had addressed the issues of shape control/maintenance of in vitro engineered human-ear-shaped cartilage. Thus, lack of applicable cell source had become a major concern that blocks clinical translation of this technology. Autologous microtia chondrocytes (MCs) and bone marrow stromal cells (BMSCs) were both promising chondrogenic cells that did not involve obvious donor site morbidity. However, limited cell availability of MCs and ectopic ossification of chondrogenically induced BMSCs in subcutaneous environment greatly restricted their applications in external ear reconstruction. The current study demonstrated that MCs possessed strong proliferation ability but accompanied with rapid loss of chondrogenic ability during passage, indicating a poor feasibility to engineer the entire ear using expanded MCs. Fortunately, the co-transplantation results of MCs and BMSCs (25% MCs and 75% BMSCs) demonstrated a strong chondroinductive ability of MCs to promote stable ectopic chondrogenesis of BMSCs in subcutaneous environment. Moreover, cell labeling demonstrated that BMSCs could transform into chondrocyte-like cells under the chondrogenic niche provided by co-cultured MCs. Most importantly, a human-ear-shaped cartilaginous tissue with delicate structure and proper elasticity was successfully constructed by seeding the mixed cells (MCs and BMSCs) into the pre-shaped biodegradable ear-scaffold followed by 12 weeks of subcutaneous implantation in nude mouse. These results may provide a promising strategy to construct stable ectopic cartilage with MCs and stem cells (BMSCs) for autologous external ear reconstruction.

  10. Potential of an injectable chitosan/starch/beta-glycerol phosphate hydrogel for sustaining normal chondrocyte function.

    Science.gov (United States)

    Ngoenkam, Jatuporn; Faikrua, Atchariya; Yasothornsrikul, Sukkid; Viyoch, Jarupa

    2010-05-31

    An injectable hydrogel for chondrocyte delivery was developed by blending chitosan and starch derived from various sources with beta-glycerol phosphate (beta-GP) in the expectation that it would retain a liquid state at room temperature and gel at raised temperatures. Rheological investigation indicated that the system consisting of chitosan derived from crab shell and corn starch at 4:1 by weight ratio (1.53%, w/v of total polymers), and 6.0% (w/v) beta-GP (C/S/GP system) exhibited the sharpest sol-gel transition at 37+/-2 degrees C. The C/S/GP hydrogel was gradually degraded by 67% within 56 days in PBS containing 0.02 mg/ml lysozyme. The presence of starch in the system increased the water absorption of the hydrogel when compared to the system without starch. SEM observation revealed to the interior structure of the C/S/GP hydrogel having interconnected pore structure (average pore size 26.4 microm) whereas the pore size of the hydrogel without starch was 19.8 microm. The hydrogel also showed an ability to maintain chondrocyte phenotype as shown by cell morphology and expression of type II collagen mRNA and protein. In vivo study revealed that the gel was formed rapidly and localized at the injection site.

  11. Natural polysaccharides promote chondrocyte adhesion and proliferation on magnetic nanoparticle/PVA composite hydrogels.

    Science.gov (United States)

    Hou, Ruixia; Nie, Lei; Du, Gaolai; Xiong, Xiaopeng; Fu, Jun

    2015-08-01

    This paper aims to investigate the synergistic effects of natural polysaccharides and inorganic nanoparticles on cell adhesion and growth on intrinsically cell non-adhesive polyvinyl alcohol (PVA) hydrogels. Previously, we have demonstrated that Fe2O3 and hydroxyapatite (nHAP) nanoparticles are effective in increasing osteoblast growth on PVA hydrogels. Herein, we blended hyaluronic acid (HA) and chondroitin sulfate (CS), two important components of cartilage extracellular matrix (ECM), with Fe2O3/nHAP/PVA hydrogels. The presence of these natural polyelectrolytes dramatically increased the pore size and the equilibrium swelling ratio (ESR) while maintaining excellent compressive strength of hydrogels. Chondrocytes were seeded and cultured on composite PVA hydrogels containing Fe2O3, nHAP and Fe2O3/nHAP hybrids and Fe2O3/nHAP with HA or CS. Confocal laser scanning microscopy (CLSM) and cell counting kit-8 (CCK-8) assay consistently confirmed that the addition of HA or CS promotes chondrocyte adhesion and growth on PVA and composite hydrogels. Particularly, the combination of HA and CS exhibited further promotion to cell adhesion and proliferation compared with any single polysaccharide. The results demonstrated that the magnetic composite nanoparticles and polysaccharides provided synergistic promotion to cell adhesion and growth. Such polysaccharide-augmented composite hydrogels may have potentials in biomedical applications.

  12. Gel-type autologous chondrocyte (Chondron™ implantation for treatment of articular cartilage defects of the knee

    Directory of Open Access Journals (Sweden)

    Chun Chung-Woo

    2010-05-01

    Full Text Available Abstract Background Gel-type autologous chondrocyte (Chondron™ implantations have been used for several years without using periosteum or membrane. This study involves evaluations of the clinical results of Chondron™ at many clinical centers at various time points during the postoperative patient follow-up. Methods Data from 98 patients with articular cartilage injury of the knee joint and who underwent Chondron™ implantation at ten Korean hospitals between January 2005 and November 2008, were included and were divided into two groups based on the patient follow-up period, i.e. 13~24-month follow-up and greater than 25-month follow-up. The telephone Knee Society Score obtained during telephone interviews with patients, was used as the evaluation tool. Results On the tKSS-A (telephone Knee Society Score-A, the score improved from 43.52 ± 20.20 to 89.71 ± 13.69 (P Conclusion Gel-type autologous chondrocyte implantation for chondral knee defects appears to be a safe and effective method for both decreasing pain and improving knee function.

  13. Ontogeny of rat chondrocyte proliferation: studies in embryo, adult and osteoarthritic (OA) cartilage

    Institute of Scientific and Technical Information of China (English)

    Madaí A GóMEZ-CAMARILLO; Juan B.KOURI

    2005-01-01

    The aim of this work was to study the ontogeny of chondrocyte cell division using embryo, adult and osteoarthritic (OA) cartilage. We searched for mitosis phases and performed a comparative evaluation of mitotic index, basic fibroblast growth factor b (FGFb), transforming growth factor β1 (TGF-β1) receptors, cyclin dependent kinase (CDK1)and Cyclin-B expression in fetal, neonate, 3, 5, 8 weeks old rats and experimental OA. Our results showed that mitosis phases were observed in all normal cartilage studied, although, we found a decrease in mitotic index in relation to tissue development. No mitosis was detected in OA cartilage. We also found a statistical significant reduction in cell number in OA cartilage, compared with the normal tissue. Furthermore, FGFb and TGF-β1 receptors diminished in relation to tissue development, and were very scarce in experimental OA. Western blot assays showed CDK-1 expression in all cases, including human-OA cartilage. Similar results were observed for Cyclin-B, except for 8 weeks, when it was not expressed. Our results suggest that cell division seems to be scarce, if not absent within the OA cartilage studied.Nevertheless, the existence of factors essential for cell division leaves open the question concerning chondrocyte proliferation in OA cartilage, which is likely to be present in the early stages of the disease.

  14. Effects of electromagnetic fields on the metabolism of lubricin of rat chondrocytes.

    Science.gov (United States)

    Wang, Wei; Li, Wenkai; Song, Mingyu; Wei, Sheng; Liu, Chaoxu; Yang, Yong; Wu, Hua

    2016-01-01

    Electromagnetic fields (EMFs) can improve pain, stiffness and physical function in osteoarthritis (OA) patients and have been proposed for the treatment of OA. However, the precise mechanisms involved in this process are still not fully understood. In the present study, we investigated the effects of exposure for different durations with 75 Hz, 2.3 mT sinusoidal EMFs (SEMFs) on the metabolism of lubricin of rat chondrocytes cultured in vitro. Our results showed that SEMFs exposure promoted lubricin synthesis in a time-dependent manner, and the expression of transforming growth factor (TGF)-β1 was also enhanced after SEMFs treatment. The up-regulation effect of the expression of lubricin under SEMF was partly reduced by SB431542, an inhibitor of TGF-RI kinase. The Smad pathway was also investigated in our study. Smad2 synthesis was higher in EMF-exposed condition than in controls, whereas no effects were observed on inhibitory Smads (Smad6 and Smad7) production. Altogether, these data suggest that SEMF exposure can promote lubricin synthesis of rat chondrocytes in a time-dependent manner and that the TGF-β/Smads signaling pathway plays a partial role.

  15. Chitosan-Coated Collagen Membranes Promote Chondrocyte Adhesion, Growth, and Interleukin-6 Secretion

    Directory of Open Access Journals (Sweden)

    Nabila Mighri

    2015-11-01

    Full Text Available Designing scaffolds made from natural polymers may be highly attractive for tissue engineering strategies. We sought to produce and characterize chitosan-coated collagen membranes and to assess their efficacy in promoting chondrocyte adhesion, growth, and cytokine secretion. Porous collagen membranes were placed in chitosan solutions then crosslinked with glutaraldehyde vapor. Fourier transform infrared (FTIR analyses showed elevated absorption at 1655 cm-1 of the carbon–nitrogen (N=C bonds formed by the reaction between the (NH2 of the chitosan and the (C=O of the glutaraldehyde. A significant peak in the amide II region revealed a significant deacetylation of the chitosan. Scanning electron microscopy (SEM images of the chitosan-coated membranes exhibited surface variations, with pore size ranging from 20 to 50 µm. X-ray photoelectron spectroscopy (XPS revealed a decreased C–C groups and an increased C–N/C–O groups due to the reaction between the carbon from the collagen and the NH2 from the chitosan. Increased rigidity of these membranes was also observed when comparing the chitosan-coated and uncoated membranes at dried conditions. However, under wet conditions, the chitosan coated collagen membranes showed lower rigidity as compared to dried conditions. Of great interest, the glutaraldehyde-crosslinked chitosan-coated collagen membranes promoted chondrocyte adhesion, growth, and interleukin (IL-6 secretion. Overall results confirm the feasibility of using designed chitosan-coated collagen membranes in future applications, such as cartilage repair.

  16. Mimicking of Chondrocyte Microenvironment Using In S