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Sample records for amniotic membrane tissue

  1. Tissue Banking in Malaysia-amniotic membrane

    International Nuclear Information System (INIS)

    Hashim bin Mohamad; Norimah binti Yusof

    1991-01-01

    Burn treatment using amniotic membranes in some of our patients initiate our own tissue bank starting with a pilot project on procurement, processing and clinical application of irradiated amniotic membrane. The irradiation of amniotic membrane was made possible by the availability of cobalt source at the Nuclear Energy Agency (UTN). With the technical help from the Inter-national Atomic Energy Agency (IAEA) we soon should be able to embark on bone bank to supply local surgeons. Thus the establishment of tissue bank at our institution will further enhance our programme which will include keratinocytes culture for burn, osteocytes culture for bone replacement as well as the use of animal skin for temporary coverage of open wounds

  2. Amniotic membrane for burn trauma

    International Nuclear Information System (INIS)

    Jamaluddin Zainol; Hasim Mohammad

    1999-01-01

    Amniotic membranes are derived from human placentae at birth. They have two layers mainly the amniotic and the chorionic surfaces which are separated by a thin layer of connective tissues. The two layers are separated during procurement, the placenta and the chorionic side are discarded and the amnion membranes are then further processed. Amnion membranes are normally procured from placentae which are normally free of infections, i.e; the mothers are antenatally screened for sexually transmitted diseases or AlDs related diseases. Intrapartum the mother should not be having chorioamnionitis or jaundice. Sometimes the amniotic membranes are acquired from fresh elective caeserian sections. After processing, the amniotic membranes are packed in two layers of polypropylene and radiated with cobalt 60 at a dose of about 25 kGy. The amniotic membranes are clinically used to cover burn surfaces especially effective for superficial or partial thickness burns. The thin membranes adhered well to the trauma areas and peeled off automatically by the second week. No change of dressing were necessary during these times because of the close adherence, there were less chance of external contamination or infections of these wounds. Due to their flexibility they are very useful to cover difference contours of the human body for example the face, body, elbows or knees. However our experience revealed that amniotic membranes are not useful for third degree bums because the membranes dissolves by the enzymes present in the wounds

  3. Biological Activity Alterations of Human Amniotic Membrane Pre and Post Irradiation Tissue Banking.

    Science.gov (United States)

    Nemr, Waleed; Bashandy, A S; Araby, Eman; Khamiss, O

    Innate immunity of Human Amniotic Membrane (HAM) and its highly active secretome that rich with various types of growth factors and anti-inflammatory substances proposed it as a promising material for many medical studies and applications. This study evaluate the biological activity of cultivated HAM pre and post tissue banking process in which freeze-dried HAM was sterilized by 25 KGray (kGy) dose of γ radiation. The HAM's antimicrobial activity, viability, growth of isolated human amniotic epithelial cells (HAECs), hematopoietic stimulation of co-cultivated murine bone marrow cells (mammalian model), scaffold efficiency for fish brain building up (non-mammalian model) and self re-epithelialization after trypsin denuding treatment were examined as supposed biological activity features. Native HAM revealed viability indications and was active to kill all tested microorganisms; 6 bacterial species (3 Gram-positive and 3 Gram-negative) and Candida albicans as a pathogenic fungus. Also, HAM activity promoted colony formation of murine hematopoietic cells, Tilapia nilotica brain fragment building-up and self re-epithelialization after trypsin treatment. In contrary, radiation-based tissue banking of HAM caused HAM cellular death and consequently lacked almost all of examined biological activity features. Viable HAM was featured with biological activity than fixed HAM prepared by irradiation tissue banking.

  4. Bacterial contamination of amniotic membrane in a tissue bank from Iran.

    Science.gov (United States)

    Aghayan, Hamid Reza; Goodarzi, Parisa; Baradaran-Rafii, Alireza; Larijani, Bagher; Moradabadi, Leila; Rahim, Fakher; Arjmand, Babak

    2013-09-01

    Human Amniotic Membrane (AM) transplantation can promote tissue healing and reduce inflammation, tissue scarring and neovascularization. Homa Peyvand Tamin (HPT) tissue bank has focused on manufacturing human cell and tissue based products including AM. The purpose of this study is to evaluate and identify bacterial contamination of AMs that is produced by HPT for several ophthalmic applications. From July 2006 to April 2011, 122 placentas from cesarean sections were retrieved by HPT after obtaining informed consent from the donors. Besides testing donor's blood sample for viral markers, microbiological evaluation was performed pre and post processing. During tissue processing, decontamination was performed by an antibiotic cocktail including; Gentamicin, Ceftriaxone and Cloxacillin. Of 271 cesarean section AM donors who were screened as potential donors, 122 were accepted for processing and assessed for microbiological contamination. Donors' age were between 21 and 41 years (Mean = 27.61 ± 0.24). More than 92% of mothers were in their first or second gravidity with full term pregnancies. The most prevalent organisms were Staphylococci species (72.53%). After processing, contamination rates markedly decreased by 84.62% (p value = 0.013). According to our results, most of bacterial contaminations were related to donation process and the contamination pattern suggests procurement team as a source. Therefore we recommend that regular training programs should be implemented by tissue banks for procurement staff. These programs should focus on improved donor screening and proper aseptic technique for tissue retrieval. We also suggest that tissue banks should periodically check the rate and types of tissue contaminations. These data help them to find system faults and to update processing methods.

  5. Tissue transglutaminase (TG-2) modified amniotic membrane: a novel scaffold for biomedical applications

    International Nuclear Information System (INIS)

    Chau, David Y S; Brown, Sheridan V; Ghaemmaghami, Amir M; Mather, Melissa L; Hutter, Victoria; Tint, Naing L; Rose, Felicity R A J; Dua, Harminder S

    2012-01-01

    The amniotic membrane (AM) is considered as a natural cell culture substrate and has occasionally been exploited in regenerative medicine especially for ocular surface reconstruction and dermal wound healing applications. However, its use is limited by its relatively weak mechanical strength, difficulty during manual handling and susceptibility to proteolytic degradation in vivo. Therefore, in this study we aimed to enhance the mechanical and biological characteristics of the AM by enzymatically cross-linking it using tissue transglutaminase (TG)—a calcium-dependent enzyme capable of forming stable ε(γ-glutamyl)lysine cross-linkages. Using a biological catalyst such as TG does not only prevent denaturation during sample preparation but also minimizes the potential of residual chemical cross-linking agents compared to alternative methodologies. Human AM, sourced from elective caesarean sectioning, were treated with TG, bovine serum albumin and/or a no-treatment control. Samples were then compared in terms of their physical and (scanning electron microscopy (SEM), transparency, mechanical strength, susceptibility to proteolytic degradation) biological characteristics (in vitro cell culture, activation of dendritic cells (DC)) and their in vivo biocompatibility/angiogenic capacity (chick chorioallantoic membrane assay). TG-treated AM exhibited enhanced mechanical strength and greater resistance to proteolytic/collagenase degradation compared to the control(s). SEM imaging of the TG-treated membrane summarized a significantly closer association and greater interconnectivity of individual collagen fibres yet it had no effect on the overall transparency of the AM. In vitro cell culture demonstrated no detrimental effect of TG-treatment on the AM in terms of cell attachment, spreading, proliferation and differentiation. Moreover, an ‘immune response’ was not elicited based on extended in vitro culture with human-monocyte-derived DC. Interestingly, the TG

  6. Amniotic Membrane Transplantation

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    Alireza Baradaran-Rafii

    2008-12-01

    Full Text Available

    The past decade has witnessed the revival of amniotic membrane transplantation (AMT in ophthalmology. The importance of amniotic membrane lies in its ability to reduce inflammation and scarring, enhance epithelialization and wound healing, and in its antimicrobial properties. Amniotic membrane has recently been used as a substrate for culturing limbal stem cells for transplantation. It has also been used extensively in corneal conditions such as neurotrophic ulcers, persistent epithelial defects, shield ulcers, microbial keratitis, band keratopathy, bullous keratopathy, and following photorefractive keratectomy and chemical injuries. Other indications for AMT include ocular surface reconstruction surgery for conjunctival pathologies such as squamous neoplasia, pterygium, and symblepharon. In this review we describe the basic structure and properties of amniotic membrane, its preparation process and its applications in ophthalmology.

  1. Evaluation of RPE65, CRALBP, VEGF, CD68, and tyrosinase gene expression in human retinal pigment epithelial cells cultured on amniotic membrane.

    Science.gov (United States)

    Akrami, Hassan; Soheili, Zahra-Soheila; Sadeghizadeh, Majid; Khalooghi, Keynoush; Ahmadieh, Hamid; Kanavi, Mojgan Rezaie; Samiei, Shahram; Pakravesh, Jalil

    2011-06-01

    The retinal pigment epithelium (RPE) plays a key role in the maintenance of the normal functions of the retina. Tissue engineering using amniotic membrane as a substrate to culture RPE cells may provide a promising new strategy to replace damaged RPE. We established a method of culturing RPE cells over the amniotic membrane as a support for their growth and transplantation. The transcription of specific genes involved in cellular function of native RPE, including RPE65, CRALBP, VEGF, CD68, and tyrosinase, were then measured using quantitative real-time PCR. Data showed a considerable increase in transcription of RPE65, CD68, and VEGF in RPE cells cultured on amniotic membrane. The amounts of CRALBP and tyrosinase transcripts were not affected. This may simply indicate that amniotic membrane restricted dedifferentiation of RPE cells in culture. The results suggest that amniotic membrane may be considered as an elective biological substrate for RPE cell culture.

  2. Treatment outcomes in the DRy Eye Amniotic Membrane (DREAM study

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    McDonald MB

    2018-04-01

    Full Text Available Marguerite B McDonald,1 Hosam Sheha,2–5 Sean Tighe,2,3 Susan B Janik,6 Frank W Bowden,7 Amit R Chokshi,8 Michael A Singer,9 Seema Nanda,10 Mujtaba A Qazi,11 Damon Dierker,12 Adam T Shupe,13 Brittany J McMurren14 1Ophthalmic Consultants of Long Island, Lynbrook, NY, USA; 2Ocular Surface Center and TissueTech, Inc., Miami, FL, USA; 3Florida International University Herbert Wertheim College of Medicine, Miami, FL, USA; 4Hofstra University School of Medicine, Hempstead, NY, USA; 5Research Institute of Ophthalmology, Cairo, Egypt; 6Solinsky Eye Care, Kensington, CT, USA; 7Bowden Eye & Associates, Jacksonville, FL, USA; 8Florida Eye Specialists, Jacksonville, FL, USA; 9Medical Center Ophthalmology Associates, San Antonio, TX, USA; 10TX Eye Institute, Houston, TX, USA; 11Pepose Vision Institute, Chesterfield, MO, USA; 12Eye Surgeons of Indiana, Indianapolis, IN, USA; 13Royo Eye Care, Marysville, CA, USA; 14Gordon and Weiss Vision Institute, San Diego, CA, USA Purpose: To evaluate the efficacy of cryopreserved amniotic membrane (CAM in reducing signs and symptoms of dry eye disease (DED in a large patient population. Methods: A retrospective chart review at 10 clinical sites was done of patients with refractory DED who received CAM and completed at least 3 months of follow-up. Data collected were demographics; medical history including previous and current ocular treatment, diagnosis, clinical presentations, comorbidity, duration and frequency of treatment with CAM; and concomitant medications. The primary outcome was the change in dry eye workshop (DEWS score after treatment. Results: A total of 97 eyes of 84 patients exhibited severe dry eye despite maximal medical treatments including topical artificial tears, cyclosporine-A, serum, antibiotics, and steroids. Patients manifested with superficial punctate keratitis (86%, filamentary keratitis (13%, exposure keratitis (19%, neurotrophic keratitis (2%, and corneal epithelial defect (7%. After CAM

  3. Microbiological viability of bovine amniotic membrane stored in glycerin 99% at room temperature for 48 months

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    Kelly Cristine de Sousa Pontes

    Full Text Available ABSTRACT The medium for storing biological tissues is of great importance for their optimal use in surgery. Glycerin has been proven efficient for storing diverse tissues for prolonged time, but the preservation of the bovine amniotic membrane in glycerin 99% at room temperature has never been evaluated to be used safely in surgical procedures. This study evaluated the preservation of 80 bovine amniotic membrane samples stored in glycerin 99% at room temperature. The samples were randomly divided evenly into four groups. Samples were microbiologically tested after 1, 6, 12 and 48 months of storage. The presence of bacteria and fungi in the samples was evaluated by inoculation on blood agar and incubation at 37 ºC for 48 hours and on Sabouraud agar at 25 ºC for 5 to 10 days. No fungal or bacterial growth was detected in any of the samples. It was concluded that glycerin is an efficient medium, regarding microbiology, for preserving pre-prepared bovine amniotic membrane, keeping the tissue free of microorganisms that grow in the media up to 48 months at room temperature.

  4. Amniotic membrane welded to contact lens by 1470-nm diode laser:a novel method for sutureless amniotic membrane transplantation

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    Rifat Rasier

    2014-12-01

    Full Text Available AIM: To avoid the side effects of the suture usage by welding amniotic membrane (AM to contact lens (CL with laser.METHODS:AM was taken from pregnant women and cleaned from blood clots with sterile phosphate-buffered physiological saline solution which included antibiotics. Stromal side of the AM was spread inside of the CL and it was welded to CL by 1470 nm diode laser. 600 μm diameter fiber tip of the laser was contacted with the epithelial side of the AM from 4 separate points. After welding excess amniotic membrane around the CL was cut with a scalpel.RESULTS:Stromal side of the AM was spread inside of the CL and then with laser fiber, different power levels and exposure times were applied on the epithelium of AM and 340 mW for seven seconds was found optimal. CL and AM attached with the spot welding effect in 4 points by touching fiber tip. CL-AM welded complex did not separated from each other while holding AM that extend beyond the CL with the help of two forceps.CONCLUSION:As a conclusion, it was aimed in this study to achieve the success of the conventional amniotic membrane transplantation (AMTwith the easiness of applying a CL and to avoid risks and side effects of corneal or conjunctival suturing. The results showed that the application of the CL–AM complex will be as easy as the application of a CL and lasts shortly.

  5. A Preliminary Study of Human Amniotic Membrane as a Potential Chondrocyte Carrier

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    L Boo

    2009-11-01

    Full Text Available PURPOSE: To investigate the feasibility of using processed human amniotic membrane (HAM to support the attachment and proliferation of chondrocytes in vitro which in turn can be utilised as a cell delivery vehicle in tissue engineering applications. METHODS: Fresh HAM obtained from patients undergoing routine elective caesarean sections was harvested, processed and dried using either freeze drying (FD or air drying (AD methods prior to sterilisation by gamma irradiation. Isolated, processed and characterised rabbit autologous chondrocytes were seeded on processed HAM and cultured for up to three weeks. Cell attachment and proliferation were examined qualitatively using inverted brightfield microscopy. RESULTS: Processed HAM appeared to allow cell attachment when implanted with chondrocytes. Although cells seeded on AD and FD HAM did not appear to attach as strongly as those seeded on glycerol preserved intact human amniotic membrane, these cells to be proliferated in cell culture conditions. CONCLUSION: Preliminary results show that processed HAM promotes chondrocyte attachment and proliferation.

  6. Roles of secretory leukocyte protease inhibitor amniotic membrane in oral wound healing

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    Elly Munadziroh

    2006-12-01

    Full Text Available Secretory Leukocyte Protease Inhibitor (SLPI is serine protease inhibitor. Secretory Leukocyte Protease Inhibitor is a protein found in secretions such as whole saliva, seminal fluid, cervical mucus, synovial fluid, breast milk, tears, and cerebral spinal fluid, as in secretions from the nose and bronchi, amniotic fluid and amniotic membrane etc. These findings demonstrate that SLPI function as a potent anti protease, anti inflammatory, bactericidal, antifungal, tissue repair, extra cellular synthesis. Impaired healing states are characterized by excessive proteolysis and often bacterial infection, leading to the hypothesis that SLPI may have a role in the process. The objectives of this article are to investigate the role of SLPI in oral inflammation and how it contributes to tissue repair in oral mucosa. The oral wound healing responses are impaired in the SLPI sufficient mice and matrix synthesis and collagen deposition are delayed. This study indicated that SLPI is a povital factor necessary for optimal wound healing.

  7. Applications of Amniotic Membrane and Fluid in Stem Cell Biology and Regenerative Medicine

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    Kerry Rennie

    2012-01-01

    Full Text Available The amniotic membrane (AM and amniotic fluid (AF have a long history of use in surgical and prenatal diagnostic applications, respectively. In addition, the discovery of cell populations in AM and AF which are widely accessible, nontumorigenic and capable of differentiating into a variety of cell types has stimulated a flurry of research aimed at characterizing the cells and evaluating their potential utility in regenerative medicine. While a major focus of research has been the use of amniotic membrane and fluid in tissue engineering and cell replacement, AM- and AF-derived cells may also have capabilities in protecting and stimulating the repair of injured tissues via paracrine actions, and acting as vectors for biodelivery of exogenous factors to treat injury and diseases. Much progress has been made since the discovery of AM and AF cells with stem cell characteristics nearly a decade ago, but there remain a number of problematic issues stemming from the inherent heterogeneity of these cells as well as inconsistencies in isolation and culturing methods which must be addressed to advance the field towards the development of cell-based therapies. Here, we provide an overview of the recent progress and future perspectives in the use of AM- and AF-derived cells for therapeutic applications.

  8. Elaboration of amniotic membrane dressing dried by air and irradiated - Peruvian experience

    International Nuclear Information System (INIS)

    Gamero, E.C.; Perez Caballero, N.

    1999-01-01

    The purpose of this work is to prepare dressings from the amniotic membrane to be used in cases of skin damage principally due to superficial and intermediate second-degree burns. The amnion is a transparent membrane that lines the chorion. It is resistant and rich in collagen. Due to these characteristics it can be well used as biological dressing as it diminishes the loss of fluids, electrolytes and proteins, it also protects the growing epithelium and adheres well to the surface of the wound, improves mobility of the patient, diminishing pain and stimulating neovascularization. The ISN-IPEN Tissue Bank promoted by IAEA has processed amniotic membrane since July 1997. Initially dressings were prepared using antibiotics, after IAEA training at the MINT of Malaysia, it is processed dried by air, lyophylized and in both presentations, sterilized by gamma-rays. Amniotic membranes are procured from Lima Maternity. Tissues must comply with VDRL, HIV, Hepatitis B and C exclusion tests. The process is held in a laminar flow hood and amnion already separated from the chorion is washed with sterile distilled water, a solution of 0.05% sodium hypochlorite, and normal saline. Then it is cut into appropriate sizes and double packed in PE films. The dressings are then carried to the Peruvian Institute of Nuclear Energy for irradiation, depending on the number of samples either irradiated with gamma-rays at the Gammacell 220 or at the Irradiation Facility located in Santa Anita. The delivered dose is 25 kGy. The product is only released if it complies with the end product quality controls. Meanwhile, microbiological tests are carried out during all the processing stages, in order to monitor the microbial load during production. In conclusion we can state that dressings prepared as above mentioned have the following advantages: not complicated preparation; reliable and safe for clinical use; diminish infection rates and days spent in the hospital; easy to storage; and can be

  9. Amniotic Tissues for the Treatment of Chronic Plantar Fasciosis and Achilles Tendinosis

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    Bruce Werber

    2015-01-01

    Full Text Available Introduction. Allogeneic amniotic tissue and fluid may be used to treat chronic plantar fasciosis and Achilles tendinosis. This innovative approach involves delivering a unique allograft of live human cells in a nonimmunogenic structural tissue matrix to treat chronic tendon injury. These tissues convey very positive regenerative attributes; procurement is performed with maternal consent during elective caesarian birth. Materials and Methods. In the present investigation all patients were unresponsive to multiple standard therapies for a minimum of 6 months and were treated with one implantation of PalinGen SportFLOW around the plantar fascia and/or around the Achilles paratenon. The patients were given a standard protocol for postimplant active rehabilitation. Results. The analogue pretreatment pain score (VAS of 8. By the fourth week after treatment, all patients had significantly reduced self-reported pain. Twelve weeks following the procedure the average pain level had reduced to only 2. No adverse reactions were reported in any of the patients. Conclusion. All patients in this study experienced heel or Achilles pain, unresponsive to standard therapy protocols. After treatment all patients noted significant pain reduction, indicating that granulized amniotic membrane and amniotic fluid can be successfully used to treat both chronic plantar fasciosis and Achilles tendinosis.

  10. The effect of amniotic membrane extract on umbilical cord blood mesenchymal stem cell expansion: is there any need to save the amniotic membrane besides the umbilical cord blood?

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    Zahra Vojdani

    2016-01-01

    Full Text Available Objective(s: Umbilical cord blood is a good source of the mesenchymal stem cells that can be banked, expanded and used in regenerative medicine.  The objective of this study was to test whether amniotic membrane extract, as a rich source of growth factors such as basic-fibroblast growth factor, can promote the proliferation potential of the umbilical cord mesenchymal stem cells. Materials and Methods: The study design was interventional. Umbilical cord mesenchymal stem cells were isolated from voluntary healthy infants from hospitals in Shiraz, Iran, cultured in the presence of basic-fibroblast growth factor and amniotic membrane extracts (from pooled - samples, and compared with control cultures. Proliferation assay was performed and duplication number and time were calculated. The expression of stem cell’s specific markers and the differentiation capacity toward osteogenic and adipogenic lineages were evaluated. Results: Amniotic membrane extract led to a significant increase in the proliferation rate and duplication number and a decrease in the duplication time without any change in the cell morphology. Both amniotic membrane extract and basic-fibroblast growth factor altered the expressing of CD44 and CD105 in cell population. Treating basic-fibroblast growth factor but not the amniotic membrane extract favored the differentiation potential of the stem cells toward osteogenic lineage. Conclusion: The amniotic membrane extract administration accelerated cell proliferation and modified the CD marker characteristics which may be due to the induction of differentiation toward a specific lineage.  Amniotic membrane extract may enhance the proliferation rate and duplication number of the stem cell through changing the duplication time.

  11. A comparative study of different amniotic membrane orientations during extraocular muscle surgery in rabbits.

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    Kassem, Rehab Rashad; El-Mofty, Randa Mohamed Abdel-Moneim; Khodeir, Mustafa Mahmoud; Hamza, Wael Mostafa

    2018-03-01

    To histopathologically compare the effect of different orientations of cryopreserved human amniotic membrane (AM) transplant during extraocular muscle surgery in rabbits. Fifty-two albino rabbit eyes underwent 4-mm resection of the superior rectus. Eyes were randomly divided into four groups. In Group C (Control group, 16 eyes) the muscle was not wrapped with amniotic membrane. In the three AM groups, cryopreserved AM was wrapped around the muscle, oriented with either its stroma (Group S, 15 eyes) or epithelium (Group E, nine eyes) towards the muscle, or folded on itself with the epithelium externally (Group F, 12 eyes). The rabbits were sacrificed and the eyes were enucleated 6 weeks after surgery. Histopathological examination was conducted for periamniotic, foreign body, scleral, and conjunctival inflammation, conjunctival vascularity, adhesions and muscle fibrosis. In all AM eyes, the AM was surrounded by periamniotic inflammation, with no adhesions detected between the muscle and surrounding tissues in the segment where the AM was present, but detected elsewhere. Adhesions were detected in all group C eyes. Foreign body inflammation was significantly less in Group C than in each of the AM groups (p  .05). Scleral inflammation was absent in all specimens. No significant differences were noted among all groups in terms of conjunctival vascularity, conjunctival inflammation, or muscle fibrosis (p > .05). All AM orientations were equally effective in preventing the development of postoperative adhesions between the extraocular muscle and surrounding tissues.

  12. Effect of Amniotic Membrane Combined with Ciprofloxacin in Curing the Primary Stages of Pseudomonal Keratitis

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    Mohammad Kazem Sharifi Yazdi

    2012-03-01

    Full Text Available Background: Keratitis caused by Pseudomonas aeruginosa is often resulted in severe corneal ulcers and perforation, which leads to losses of vision. Human amniotic membrane (HAM forms the inner wall of the membranous sac which surrounds and protects the embryo during gestation. The purpose of this study was to evaluate the effectiveness of the amniotic membrane's healing in rabbits with pseudomonas keratitis.Methods: In total 14 rabbits divided in 2 groups of: 1 as Control and 2 as experimental amniotic membrane combined with ciprofloxacin. A 0.05 ml suspension of Pseudomonas aeruginosa ATCC 27853 was injected into rabbit’s corneal stroma, with no interference in control group. In the second group, the amniotic membrane in pieces of 1.5 × 1.5 cm transplanted to the entire corneal surface by eight interrupted 10.0 nylon sutures. In the first day ciprofloxacin drop was injected to the second group every 30 minutes and through second to seventh days every 2 hours. The results of perforation in cornea and the amount of infiltration were registered.Results: The results showed that amniotic membrane transplantation (AMT + ciprofloxacin group had 0% perforation and the control group 85.6%. Average infiltrations were 5 mm in AMT + ciprofloxacin groups and 23.75 mm in control.Conclusion: The use of amniotic membrane with ciprofloxacin was effective in prevention of cornea perforation and controlling the process of pseudomonal keratitis remission. The improvement of inflammation rapidly happened in ciprofloxacin + AMT group.

  13. Clinical outcome of combined conjunctival autograft transplantation and amniotic membrane transplantation in pterygium surgery

    OpenAIRE

    Tejsu Malla; Jing Jiang; Kai Hu

    2018-01-01

    AIM: To compare long-term outcome of primary and recurrent pterygium surgery with three different techniques: combined conjunctival autograft and overlay amniotic membrane transplantation (CAT with AMT), conjunctival autograft transplantation (CAT) alone and amniotic membrane transplantation (AMT) alone. METHODS: In this retrospective study, 142 eyes of 142 pterygium patients (104 primary, 38 recurrent) who underwent CAT (group A), AMT (group B) or CAT with AMT (group C) respectively follo...

  14. Developmental mechanisms of the tympanic membrane in mammals and non-mammalian amniotes.

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    Takechi, Masaki; Kitazawa, Taro; Hirasawa, Tatsuya; Hirai, Tamami; Iseki, Sachiko; Kurihara, Hiroki; Kuratani, Shigeru

    2016-01-01

    The tympanic membrane is a thin layer that originates from the ectoderm, endoderm, and mesenchyme. Molecular-genetic investigations have revealed that interaction between epithelial and mesenchymal cells in the pharyngeal arches is essential for development of the tympanic membrane. We have recently reported that developmental mechanisms underlying the tympanic membrane seem to be different between mouse and chicken, suggesting that the tympanic membrane evolved independently in mammals and non-mammalian amniotes. In this review, we summarize previous studies of tympanic membrane formation in the mouse. We also discuss its formation in amniotes from an evolutionary point of view. © 2015 Japanese Teratology Society.

  15. Human Amniotic Membrane Dressing: an Excellent Method for Outpatient Management of Burn Wounds

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    Ali Akbar Mohammadi

    2009-03-01

    Full Text Available Background: Burns are among the most common traumas indeveloping countries, which consume large amounts of medicalresources. It is important to find an appropriate materialfor dressing of burn wounds that improves healing and is readilyavailable, easily applicable, and economical.Methods: In a single-blind randomized controlled clinicaltrial from March to October 2006, 211 patients with less than20% burn were enrolled into two groups. The first group contained104 patients with average burn of 11.90± 3.80% of totalbody surface area (TBSA for whom amnion dressing wasused. The second group composed of 107 patients with averageburn of 12.30± 4.14% of TBSA treated with routine silversulfadiazine dressing.Results: Amniotic membrane usage was accompanied by accelerationin wound healing, less need for skin graft, and lesspain. The mean healing time in superficial parts of burnwounds in the amnion group was significantly shorter than thecontrol group (9.50±2.13 v 14.30±2.60 days; P value < 0.01.The extent of the wound with granulation tissue which neededskin graft was less in the amnion group (2.10 ± 2.21% v 4.20±1.44%; P value < 0.01.Conclusion: Widespread use of amniotic membrane dressingis recommended for limited burn wound management.

  16. Amniotic membrane transplantation for reconstruction of corneal epithelial surface in cases of partial limbal stem cell deficiency.

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    Sangwan Virender

    2004-01-01

    Full Text Available Purpose: To assess the efficacy of amniotic membrane for treatment of partial limbal stem cell deficiency (LSCD. Methods: Medical records of four patients with partial LSCD who underwent pannus resection and amniotic membrane transplantation (AMT were reviewed for ocular surface stability and improvement in visual acuity. Clinico-histopathological correlation was done with the resected pannus tissue. Results: All the eyes exhibited stable corneal epithelial surface by an average of 7 weeks postoperatively with improvement in subjective symptoms. Best corrected visual acuity improved from preoperative (range: 6/9p-6/120 to postoperative (range: 6/6p-6/15 by an average of 4.5 lines on Snellen visual acuity charts. Histopathological examination of excised tissue showed features of conjunctivalisation. Conclusion: Amniotic membrane transplantation appears to be an effective means of reconstructing the corneal epithelial surface and for visual rehabilitation of patients with partial limbal stem cell deficiency. It may be considered as an alternative primary procedure to limbal transplantation in these cases.

  17. Bone density of defects treated with lyophilized amniotic membrane versus colagen membrane: a tomographic and histomorfogenic study in a rabbi´s femur.

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    Liz Ríos

    2014-09-01

    Full Text Available The aim of this study was to compare the bone density of bone defects treated with lyophilizated amniotic membrane (LAM and collagen Membrane (CM, at 3 and 5 weeks. Two bone defects of 4mm in diameter and 6mm deep were created in left distal femoral diaphysis of New Zealand rabbits (n=12. The animals were randomly divided into 2 groups. One of the defects was covered with lyophilized amniotic membrane (Rosa Chambergo Tissue Bank/National Institute of Child Health-IPEN, Lima, Peru or collagen Membrane (Dentium Co, Seoul, Korea. The second was left uncovered (NC. The rabbits were killed after 3 and 5 weeks (3 rabbits/period. The results showed a high bone density and repair of the defect by new bone. The tomographic study revealed that the bone density of the defects treated with LAM at 3 weeks was equivalent to the density obtained with CM and higher density compared with NC (p0.05. The results show that lyophilizated amniotic membrane provides bone density equal or higher to the collagen membrane.

  18. Cat amniotic membrane multipotent cells are nontumorigenic and are safe for use in cell transplantation

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    Vidane AS

    2014-08-01

    Full Text Available Atanasio S Vidane,1 Aline F Souza,1 Rafael V Sampaio,1 Fabiana F Bressan,2 Naira C Pieri,1 Daniele S Martins,2 Flavio V Meirelles,2 Maria A Miglino,1 Carlos E Ambrósio2 1Department of Surgery, Faculty of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, Brazil; 2Department of Veterinary Medicine, Faculty of Animal Sciences and Food Engineering, University of São Paulo, São Paulo, Brazil Abstract: Amnion-derived mesenchymal stem cells (AMSCs are multipotent cells with an enhanced ability to differentiate into multiple lineages. AMSCs can be acquired through noninvasive methods, and therefore are exempt from the typical ethical issues surrounding stem cell use. The objective of this study was to isolate and characterize AMSCs from a cat amniotic membrane for future application in regenerative medicine. The cat AMSCs were harvested after mechanical and enzymatic digestion of amnion. In culture medium, the cat AMSCs adhered to a plastic culture dish and displayed a fibroblast-like morphology. Immunophenotyping assays were positive for the mesenchymal stem cell-specific markers CD73 and CD90 but not the hematopoietic markers CD34, CD45, and CD79. Under appropriate conditions, the cat AMSCs differentiated into osteogenic, chondrogenic, and adipogenic cell lineages. One advantage of cat AMSCs was nonteratogenicity, assessed 4 weeks post injection of undifferentiated AMSCs into immunodeficient mice. These findings suggest that cat amniotic membranes may be an important and useful source of mesenchymal stem cells for clinical applications, especially for cell or tissue replacement in chronic and degenerative diseases. Keywords: amnion, cats, cell differentiation, fetal membranes, mesenchymal cells

  19. Some biological properties of the human amniotic membrane interferon

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    P. C. P. Ferreira

    1992-03-01

    Full Text Available Human amniotic interferon was investigated to define the species specificity of its antiviral action and compare its anti-cellular and NK cell stimulating activities with those of other human interferons. The antiviral effect was titrated in bovine (RV-IAL and monkey (VERO cells. Amniotic interferon exhibited, in bovine cells, 5% of the activity seen in monkey cells, while alpha interferon displayed 200%. No effect was detected with either beta or gamma interferon in bovine cells. Daudi cells were exposed to different concentrations of various interferons and the cell numbers were determined. The anticellular effect of the amniotic interferon reached its peak on the third day of incubation. Results suggested a higher activity for alpha and gamma interferons and a lower activity for beta when compared to amniotic interferon. Using total mononuclear cells as effector cells and K 562 as target cell in a 51Cr release assay, it was demonstrated that low concentrations of amniotic interferon consistently stimulated NK cell activity in cells derived from several donors, the results indicating a higher level of activity with this interferon than with alpha and beta interferons.

  20. Treatment of ligneous conjunctivitis with amniotic membrane transplantation and topical cyclosporine

    Science.gov (United States)

    Tok, Ozlem Yalcin; Kocaoglu, Fatma Akbas; Tok, Levent; Burcu, Ayse; Ornek, Firdevs

    2012-01-01

    Ligneous conjunctivitis (LC) is a rare form of bilateral chronic recurrent disease in which thick membranes form on the palpebral conjunctiva and other mucosal sites. We report the clinical features and describe the management of two cases. Case 1 was an 8-month-old patient with bilateral membranous conjunctivitis. Case 2 was a 5-year-old patient with unilateral membranous conjunctivitis, esotropia, mechanical ptosis and complicated cataract, and had been treated with a number of medications. Histological investigation of the membrane in both cases showed LC. Treatments with amniotic membrane transplantation and institution of topical cyclosporine have shown good response. There has been complete resolution of the membranes with no recurrence at the end of 40- and 28-month follow-ups, respectively. No treatment related side effects were seen. Thus, it appears that amniotic membrane transplantation and topical cyclosporine are effective alternatives for the treatment of LC. PMID:23202401

  1. INTERLEUKIN-6 TRANS-SIGNALING SYSTEM IN INTRA-AMNIOTIC INFLAMMATION, PRETERM BIRTH AND PRETERM PREMATURE RUPTURE OF THE MEMBRANES

    Science.gov (United States)

    Lee, Sarah Y.; Buhimschi, Irina A.; Dulay, Antonette T.; Ali, Unzila A.; Zhao, Guomao; Abdel-Razeq, Sonya S.; Bahtiyar, Mert O.; Thung, Stephen F.; Funai, Edmund F.; Buhimschi, Catalin S.

    2013-01-01

    Classic IL-6 signaling is conditioned by the transmembrane receptor (IL-6R) and homodimerization of gp130. During trans-signaling, IL-6 binds to soluble IL-6R (sIL-6R) enabling activation of cells expressing solely gp130. Soluble gp130 (sgp130) selectively inhibits IL-6 trans-signaling. To characterize amniotic fluid IL-6 trans-signaling molecules (IL-6, sIL-6R, sgp130) in normal gestations and pregnancies complicated by intra-amniotic inflammation (IAI) we studied 301 women during second trimester (n=39), third trimester (n=40) and preterm labor with intact (n=131, 85 IAI negative & 46 IAI positive) or preterm premature rupture of membranes (PPROM: n=91, 61 IAI negative & 30 IAI positive). ELISA, Western blotting and RT-PCR were used to investigate amniotic fluid, placenta and amniochorion for protein and mRNA expression of sIL-6R, sgp130, IL-6R and gp130. Tissues were immunostained for IL-6R, gp130, CD15+ (polymorphonuclear) and CD3+ (T-cell) inflammatory cells. The ability of sIL-6R and sgp130 to modulate basal and LPS-stimulated release of amniochorion matrix-metalloprotease-9 (MMP-9) was tested ex-vivo. We showed that in physiologic gestations amniotic fluid sgp130 decreases toward term. Amniotic fluid IL-6 and sIL-6R were elevated in IAI whereas sgp130 was decreased in PPROM. Our results suggested that fetal membranes are the probable source of amniotic fluid sIL-6R and sgp130. Immunohistochemistry and RT-PCR revealed increased IL-6R and decreased gp130 expression in amniochorion of women with IAI. Ex-vivo, sIL-6R and LPS augmented amniochorion MMP-9 release whereas sgp130 opposed this effect. We conclude that IL-6 trans-signaling molecules are physiologic constituents of the amniotic fluid regulated by gestational age and inflammation. PPROM likely involves functional loss of sgp130. PMID:21282511

  2. Clinical analysis of amniotic membrane patches and grafts for acute ocular surface burn

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    Lin Li

    2015-01-01

    Full Text Available AIM: To investigate the effect and value of amniotic membrane patches and grafts for acute ocular surface burn at different degrees.METHODS: A retrospective analysis of 28 cases(28 eyesaffected by ocular chemical or thermal burn with different degree were included in our hospital from March 2007 to March 2012. Amniotic membrane patched was undergone in 13 eyes with fresh amnion that the patients corneal burns degree Ⅱ or Ⅲ with partial limbal buns at degree Ⅳ. Amniotic membrane grafts was performed in 15 eyes with fresh amnion that the patients all corneal burns at degree Ⅲ with the whole limbal necrosis without severe eyelid defect. The follow-up time ranged 6~24mo. The postoperative visual acuity, the condition of amniotic membrane transplant, renovation of cornea and complications were observed. RESULTS: Postoperative corrected visual acuity was improved in 20 eyes(71%, it was not changed in 5 eyes(18%, the visual acuity declined in 3 eyes(11%. The amniotic membrane survived in 23 eyes and the survival rate was up to 82%. The cornea of 4 eyes recovered to transparent, nebula emceed in 8 eyes eventually, corneal macula emerged in 10 eyes, 4 eyes ended up with leukoma, 2 eyes developed corneal melting after therapy, then received lamellar keratoplasty. Corneal surface become epithelization after amnion patches or grafts, but any of them have recurrent epithelial erosion, and become stable epithalization after repeat operation.CONCLUSION: Amniotic membrane patches and grafts is an effective method to deal with acute ocular surface burn.

  3. Biomechanical assays amniotic membrane preserved in glycerol correlating with optical coherence tomography (OCT) and thermal gravimetric analysis (TG)

    International Nuclear Information System (INIS)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Freitas, Anderson Z.; Mathor, Monica B.

    2013-01-01

    Amnion or amniotic membranes (AM) are interchangeable terms used in the literature being internal part of the fetal membranes, non-vascular and multicellular tissue. The amnion has been widely used as a graft ophthalmic surgical as well as carrier substrate stem cell tissue equivalent for ocular surface reconstruction. The AM reduces scar formation and inflammation on the ocular surface, promotes epithelization also been used as a biological bandage covering the wound or burns, reducing dehydration and allowing regeneration of these areas. The amnion has usually 0.02 to 0.5 mm thick and consists of five subsequent layers: epithelium, basement membrane, compact layer, fibroblast layer and spongy layer. The mechanical strength from the membrane structure as well as the elasticity are factors attractive to the use of amnion as a surgical graft. Higher levels of rigidity and strength may improve the graft resistance necessary to resist the stress induced during growth of the new tissue formed. The amniotic membrane is obtained at elective caesarean section and subsequently, under sterile conditions, sectioned and separated from chorion and placenta, and free blood clots. The serological tests are done at the time of collection of tissue and 6 months after delivery to confirm the results. There are different methods for storing MA in tissue banks as fresh, high concentrations of glycerol, among others. The use of fresh membrane has some limitations due to the need to rapid use and high risk of contamination, however the amniotic membrane in glycerol has antiviral and antibacterial property which is dependent on the concentration, time and temperature. The AM used in transplants must be sterile to prevent the transmission of any disease. Although sterilization by radiation is an effective procedure, it can interfere on the membrane structure. Thus, verification of potential changes caused by ionizing radiation in amnion was made using the tensile test by calculating the

  4. Biomechanical assays amniotic membrane preserved in glycerol correlating with optical coherence tomography (OCT) and thermal gravimetric analysis (TG)

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Fernando Augusto N.; Santin, Stefany P.; Martino Junior, Antonio C.; Machado, Luci Diva B.; Freitas, Anderson Z.; Mathor, Monica B., E-mail: fernandonevessoares@yahoo.com.br [Instituto de Pesquisas Energetias Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2013-07-01

    Amnion or amniotic membranes (AM) are interchangeable terms used in the literature being internal part of the fetal membranes, non-vascular and multicellular tissue. The amnion has been widely used as a graft ophthalmic surgical as well as carrier substrate stem cell tissue equivalent for ocular surface reconstruction. The AM reduces scar formation and inflammation on the ocular surface, promotes epithelization also been used as a biological bandage covering the wound or burns, reducing dehydration and allowing regeneration of these areas. The amnion has usually 0.02 to 0.5 mm thick and consists of five subsequent layers: epithelium, basement membrane, compact layer, fibroblast layer and spongy layer. The mechanical strength from the membrane structure as well as the elasticity are factors attractive to the use of amnion as a surgical graft. Higher levels of rigidity and strength may improve the graft resistance necessary to resist the stress induced during growth of the new tissue formed. The amniotic membrane is obtained at elective caesarean section and subsequently, under sterile conditions, sectioned and separated from chorion and placenta, and free blood clots. The serological tests are done at the time of collection of tissue and 6 months after delivery to confirm the results. There are different methods for storing MA in tissue banks as fresh, high concentrations of glycerol, among others. The use of fresh membrane has some limitations due to the need to rapid use and high risk of contamination, however the amniotic membrane in glycerol has antiviral and antibacterial property which is dependent on the concentration, time and temperature. The AM used in transplants must be sterile to prevent the transmission of any disease. Although sterilization by radiation is an effective procedure, it can interfere on the membrane structure. Thus, verification of potential changes caused by ionizing radiation in amnion was made using the tensile test by calculating the

  5. Assessment of amniotic and polyurethane membrane dressings in the treatment of burns.

    Science.gov (United States)

    Adly, O A; Moghazy, A M; Abbas, A H; Ellabban, A M; Ali, O S; Mohamed, B A

    2010-08-01

    As allograft and xenografts are not available in Islamic countries, amniotic membrane seems to be an effective alternative in the management of deep burns. Its proven bioactivities and modest price suggest that it might be superior to synthetic dressings. Forty-six patients were enrolled in this randomized, controlled clinical trial conducted in the Burn Unit at Suez Canal University Hospital, Ismailia, Egypt. All age groups and both gender were included in the study. Only patients with less than 50% total body surface area burned were included, thus minimizing the dropouts in both groups. All were either second or third degree. These patients were randomly assigned either to group I: amniotic membrane (Biomembrane) dressing, or group II: polyurethane membrane (Tegaderm) dressing. Those in group I demonstrated a significantly lower rate of infection and required less frequent dressing changes than those in group II. They also sustained less electrolyte and albumin loss. The rate of healing in the amniotic membrane group was significantly faster than in the polyurethane group. Furthermore, pain was significantly less when Biomembrane was used. Based on these findings, we recommend the use of lyophilized gamma-irradiated amniotic membrane as an effective alternative for allograft and xenografts in Islamic countries and the Jewish population.

  6. Human Amniotic Membrane-Derived Products in Sports Medicine: Basic Science, Early Results, and Potential Clinical Applications.

    Science.gov (United States)

    Riboh, Jonathan C; Saltzman, Bryan M; Yanke, Adam B; Cole, Brian J

    2016-09-01

    Amniotic membrane (AM)-derived products have been successfully used in ophthalmology, plastic surgery, and wound care, but little is known about their potential applications in orthopaedic sports medicine. To provide an updated review of the basic science and preclinical and clinical data supporting the use of AM-derived products and to review their current applications in sports medicine. Systematic review. A systematic search of the literature was conducted using the Medline, EMBASE, and Cochrane databases. The search term amniotic membrane was used alone and in conjunction with stem cell, orthopaedic, tissue engineering, scaffold, and sports medicine. The search identified 6870 articles, 80 of which, after screening of the titles and abstracts, were considered relevant to this study. Fifty-five articles described the anatomy, basic science, and nonorthopaedic applications of AM-derived products. Twenty-five articles described preclinical and clinical trials of AM-derived products for orthopaedic sports medicine. Because the level of evidence obtained from this search was not adequate for systematic review or meta-analysis, a current concepts review on the anatomy, physiology, and clinical uses of AM-derived products is presented. Amniotic membranes have many promising applications in sports medicine. They are a source of pluripotent cells, highly organized collagen, antifibrotic and anti-inflammatory cytokines, immunomodulators, and matrix proteins. These properties may make it beneficial when applied as tissue engineering scaffolds, improving tissue organization in healing, and treatment of the arthritic joint. The current body of evidence in sports medicine is heavily biased toward in vitro and animal studies, with little to no human clinical data. Nonetheless, 14 companies or distributors offer commercial AM products. The preparation and formulation of these products alter their biological and mechanical properties, and a thorough understanding of these

  7. Amniotic membrane transplantation ineffective as additional therapy in patients with aggressive Mooren's ulcer.

    Science.gov (United States)

    Schallenberg, Maurice; Westekemper, Henrike; Steuhl, Klaus-Peter; Meller, Daniel

    2013-12-17

    Mooren's ulcer is a severe ulcerative inflammation of the cornea. The exact pathogenesis remains unclear. Therefore many therapies of Mooren's ulcer are recommended in literature. To shed more light on the ongoing question of optimal treatment of severe progressive Mooren's ulcer, we here report on a retrospective case series of patients treated with systemic immunosuppressive therapy and additional amniotic membrane transplantation. Medical records from seven patients (eleven eyes), 4 male and 3 female, with severe progressive Mooren's ulcer were analysed retrospectively. The mean follow up was 88.4 ± 80.8 months (range 12-232 month). A HLA-typing was performed in all patients. A systemic immunosuppressive therapy was administered in all patients. The amniotic membrane was transplanted after the base of the ulcer was resected. Multiple amniotic membrane transplantations were necessary in six patients. The visual outcome of all patients was poor. No patient achieved a visual acuity better than 20/630 Snellen chart. Five patients were positive for HLA-DQ2 and four patients were positive for HLA-DR17(3). The aggressive and highly inflammatory form of Mooren's ulcer is difficult to treat and the progression of the disease is hard to influence positively even under systemic immunosuppressive therapy. Therefore, the main intention of therapy is to achieve a stable epithelialized corneal surface without the risk of perforation. Amniotic membrane transplantation is not able to cure severe forms of Mooren's ulcer. However it supports the immunosuppressive therapy in acute situations as in critical corneal thinning.

  8. Effect of the Human Amniotic Membrane on Liver Regeneration in Rats

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    Mesut Sipahi

    2015-01-01

    Full Text Available Introduction. Operations are performed for broader liver surgery indications for a better understanding of hepatic anatomy/physiology and developments in operation technology. Surgery can cure some patients with liver metastasis of some tumors. Nevertheless, postoperative liver failure is the most feared complication causing mortality in patients who have undergone excision of a large liver mass. The human amniotic membrane has regenerative effects. Thus, we investigated the effects of the human amniotic membrane on regeneration of the resected liver. Methods. Twenty female Wistar albino rats were divided into control and experimental groups and underwent a 70% hepatectomy. The human amniotic membrane was placed over the residual liver in the experimental group. Relative liver weight, histopathological features, and biochemical parameters were assessed on postoperative day 3. Results. Total protein and albumin levels were significantly lower in the experimental group than in the control group. No difference in relative liver weight was observed between the groups. Hepatocyte mitotic count was significantly higher in the experimental group than in the control group. Hepatic steatosis was detected in the experimental group. Conclusion. Applying the amniotic membrane to residual liver adversely affected liver regeneration. However, mesenchymal stem cell research has the potential to accelerate liver regeneration investigations.

  9. Nukbone® promotes proliferation and osteoblastic differentiation of mesenchymal stem cells from human amniotic membrane

    International Nuclear Information System (INIS)

    Rodríguez-Fuentes, Nayeli; Rodríguez-Hernández, Ana G.; Enríquez-Jiménez, Juana; Alcántara-Quintana, Luz E.; Fuentes-Mera, Lizeth; Piña-Barba, María C.; Zepeda-Rodríguez, Armando

    2013-01-01

    Highlights: •Nukbone showed to be a good scaffold for adhesion, proliferation and differentiation of stem cells. •Nukbone induced osteoblastic differentiation of human mesenchymal stem cells. •Results showed that Nukbone offer an excellent option for bone tissue regeneration due to properties. -- Abstract: Bovine bone matrix Nukbone® (NKB) is an osseous tissue-engineering biomaterial that retains its mineral and organic phases and its natural bone topography and has been used as a xenoimplant for bone regeneration in clinics. There are not studies regarding its influence of the NKB in the behavior of cells during the repairing processes. The aim of this research is to demonstrate that NKB has an osteoinductive effect in human mesenchymal stem cells from amniotic membrane (AM-hMSCs). Results indicated that NKB favors the AM-hMSCs adhesion and proliferation up to 7 days in culture as shown by the scanning electron microscopy and proliferation measures using an alamarBlue assay. Furthermore, as demonstrated by reverse transcriptase polymerase chain reaction, it was detected that two gene expression markers of osteoblastic differentiation: the core binding factor and osteocalcin were higher for AM-hMSCs co-cultured with NKB in comparison with cultivated cells in absence of the biomaterial. As the results indicate, NKB possess the capability for inducing successfully the osteoblastic differentiation of AM-hMSC, so that, NKB is an excellent xenoimplant option for repairing bone tissue defects

  10. Nukbone® promotes proliferation and osteoblastic differentiation of mesenchymal stem cells from human amniotic membrane

    Energy Technology Data Exchange (ETDEWEB)

    Rodríguez-Fuentes, Nayeli; Rodríguez-Hernández, Ana G. [Depto. Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Mexico City 04510 (Mexico); Enríquez-Jiménez, Juana [Depto. Biología de la Reproducción, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán (INCMNSZ), México City 14000 (Mexico); Alcántara-Quintana, Luz E. [Subd. de Investigación, Centro Nacional de la Transfusión Sanguínea, Secretaria de Salud, Mexico City 07370 (Mexico); Fuentes-Mera, Lizeth [Depto. Biología Molecular e Histocompatibilidad, Hospital General “Dr. Manuel Gea González”, México City 4800 (Mexico); Piña-Barba, María C. [Depto. Materiales Metálicos y Cerámicos, Instituto de Investigaciones en Materiales, Universidad Nacional Autónoma de México (UNAM), México City 04510 (Mexico); Zepeda-Rodríguez, Armando [Depto. Biología Celular y Tisular, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), México City 04510 (Mexico); and others

    2013-05-10

    Highlights: •Nukbone showed to be a good scaffold for adhesion, proliferation and differentiation of stem cells. •Nukbone induced osteoblastic differentiation of human mesenchymal stem cells. •Results showed that Nukbone offer an excellent option for bone tissue regeneration due to properties. -- Abstract: Bovine bone matrix Nukbone® (NKB) is an osseous tissue-engineering biomaterial that retains its mineral and organic phases and its natural bone topography and has been used as a xenoimplant for bone regeneration in clinics. There are not studies regarding its influence of the NKB in the behavior of cells during the repairing processes. The aim of this research is to demonstrate that NKB has an osteoinductive effect in human mesenchymal stem cells from amniotic membrane (AM-hMSCs). Results indicated that NKB favors the AM-hMSCs adhesion and proliferation up to 7 days in culture as shown by the scanning electron microscopy and proliferation measures using an alamarBlue assay. Furthermore, as demonstrated by reverse transcriptase polymerase chain reaction, it was detected that two gene expression markers of osteoblastic differentiation: the core binding factor and osteocalcin were higher for AM-hMSCs co-cultured with NKB in comparison with cultivated cells in absence of the biomaterial. As the results indicate, NKB possess the capability for inducing successfully the osteoblastic differentiation of AM-hMSC, so that, NKB is an excellent xenoimplant option for repairing bone tissue defects.

  11. Evaluation of a dental pulp-derived cell sheet cultured on amniotic membrane substrate.

    Science.gov (United States)

    Honjo, Ken-ichi; Yamamoto, Toshiro; Adachi, Tetsuya; Amemiya, Takeshi; Mazda, Osam; Kanamura, Narisato; Kita, Masakazu

    2015-01-01

    Mesenchymal stem cells (MSC) are transplanted for periodontal tissue regeneration, and the periodontal ligament (PDL) is regenerated using a cultured cell sheet. This cultured cell sheet is prepared using PDL-derived cells, growth factors, and amniotic membrane (AM). Dental pulp (DP)-derived cells can be easily obtained from extracted wisdom teeth, proliferate rapidly, and are less susceptible to bacterial infection than PDL-derived cells. Thus, to prepare a novel cell sheet, DP-derived cells were cultured on AM as a culture substrate for immunohistochemical examination. Wisdom teeth extracted from three adults were cut along the cement-enamel border. DP tissue was collected, minced, and primarily cultured. After three or four passage cultures, DP-derived cells were cultured on AM, followed by hematoxylin-eosin (H-E) and immunofluorescence staining. DP-derived cells cultured on AM formed a layered structure. Cells positive for vimentin, Ki-67, ZO-1, desmoplakin, CD29, 44, 105 or 146, STRO-1, collagen IV or VII or laminin 5 or α5 chain were localized. DP-derived cells proliferated on AM, while retaining the properties of DP, which allowed the cultured cell sheet to be prepared. In addition, the cultured cell sheet contained MSC, which suggests its potential application in periodontal tissue regeneration.

  12. Construction and characterization of human oral mucosa equivalent using hyper-dry amniotic membrane as a matrix.

    Science.gov (United States)

    Qi, Fangfang; Yoshida, Toshiko; Koike, Takeshi; Aizawa, Hitoshi; Shimane, Tetsu; Li, Yinghui; Yamada, Shinichi; Okabe, Motonori; Nikaido, Toshio; Kurita, Hiroshi

    2016-05-01

    Human amniotic membrane(HAM) as a graft material has been used in various fields. Hyper-dry amniotic membrane (HD-AM) is a novel dried amniotic membrane that is easy to handle and can be preserved at room temperature without time limitation. The purpose of this study was to investigate the useful properties of HD-AM in reconstruction of the oral mucosa. Human oral keratinocytes were isolated and seeded on HD-AM in serum-free culture system. Oral mucosa equivalent (OME) was developed and transplanted onto full-thickness wound on athymic mice. The wound healing was analyzed and the OME both before and after transplantation was analyzed with hematoxylin-eosin staining and immunohistochemical staining for Cytokines 10 (CK10), Cytokines 16 (CK16), and Ivolucrin (IVL). Oral keratinocytes spread and proliferated well on HD-AM. Two weeks after air-lifting, OME had formed with good differentiation and morphology. We confirmed immunohistochemically that the expression of CK10 was positive in all suprabasal layers, as was CK16 in the upper layers, while IVL was present in all cell layers. Three weeks after transplantation to athymic mice, the newly generated tissue had survived well with the smallest contraction. The epithelial cells of newly generated tissue expressed CK10 throughout in all suprabasal layers, IVL was mainly in the granular layer, and CK16 positive cells were observed in all spinous layer and granular layer but were not expressed in the mouse skin, all of which were similar to native gingival mucosa. The OME with HD-AM as a matrix revealed a good morphology and stable wound healing. This study demonstrates that HD-AM is a useful and feasible biomaterial for oral mucosa reconstruction. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Clinical application of amniotic membranes on a patient with epidermolysis bullosa.

    Science.gov (United States)

    Martínez Pardo, M E; Reyes Frías, M L; Ramos Durón, L E; Gutiérrez Salgado, E; Gómez, J C; Marín, M A; Luna Zaragoza, D

    1999-01-01

    The case of a patient with dystrophic epidermolysis bullosa treated with radiosterilised amniotic membranes is presented. The disorder is a congenital disease characterised by a poor desmosomal junction in the keratinocyte membrane. After proper donor screening, amnios were collected at Hospital Central Sur de Alta Especialidad (HCSAE), PEMEX and microbiological analysis was performed at Universidad Nacional Autónoma de México, FQUNAM, (Biology Dept. of the Chemistry Faculty, National Autonomous University of Mexico), before and after radiation sterilisation. Processing, packaging and sterilisation were performed at Instituto Nacional de Investigaciones Nucleares, ININ, (National Nuclear Research Institute). The patient, a ten-year-old boy with severe malnutrition, extensive loss of skin and pseudomonad infection in the whole body, was treated with gentle debridement in a Hubbard bath. Later amnion application was performed with sterilised amnios by using two different processes, in one of which the amnion was sterilised with paracetic acid, preserved in glycerol, kindly donated by the German Institute for Tissue and Cell Replacement and applied by Dr. Johannes C. Bruck, IAEA visiting expert, and the other amnion was processed at ININ: air dried and sterilised by gamma radiation at dose of 30 kGy. After spontaneous epithelisation was successfully promoted for seven days, the pain was alleviated and mobility was improved in a few hours and the patient's general condition was so improved that in a month he was discharged. Unfortunately, because this disease is revertive and has malignant degeneration, the prognosis is not good.

  14. Amniotic membranes as prosthetic material: experimental utilization data of a rat model.

    Science.gov (United States)

    Zachariou, Z

    1997-10-01

    Prosthetic materials are applied for closing big tissue defects, the repair of traumatized organs, or hernias. Because nonabsorbable synthetic materials are rigid, possess a defined and unchangeable size, and foreign body reaction (FBR) may occur, biological materials may be an alternative. In experimental studies in rats the authors implanted the fetal parts of the human amniotic membranes and examined the utilization and FBR induced in a standardized model. In addition amnion (AM) was combined with vicryl-net (VN) for higher implant stability. Fifteen, 30 and 90 days after implantation, macroscopic appearance was examined, and light microscopy and immunohistology testing of the specimens were performed. Adhesions to parenchymal organs and omentum were present irrespective of the side facing the abdominal cavity. AM induced a rapid FBR, which diminished with time. Chorion (CH) and parts of the AM were resorbed within the examined period after infiltration with recipient cells and neovascularisation. The combined implant, AM, and VN showed best results because disadvantages of one material could be compensated for by the advantages of the other. The studies show that AM, in its anatomic definition, combined with VN proves to be a safe and reliable prosthetic material for the use in tissue defects.

  15. Annexin A2 in amniotic fluid: correlation with histological chorioamnionitis, preterm premature rupture of membranes, and subsequent preterm delivery.

    Science.gov (United States)

    Namba, Fumihiko; Ina, Shihomi; Kitajima, Hiroyuki; Yoshio, Hiroyuki; Mimura, Kazuya; Saito, Shigeru; Yanagihara, Itaru

    2012-01-01

    The aim of this study was to determine whether amniotic fluid levels of annexin A2, a phospholipid-binding protein that is abundant in amnion and regulates fibrin homeostasis, are associated with histological chorioamnionitis, preterm premature rupture of the membranes, and subsequent preterm delivery. Amniotic fluid was obtained from 55 pregnant women with preterm labor and/or preterm premature rupture of the membranes before 32weeks of gestation, and amniotic fluid levels of annexin A2 were measured with a sandwich enzyme-linked immunosorbent assay. Amniotic fluid levels of annexin A2 in patients with histological chorioamnionitis was higher than that in the remainder (P=0.053), whereas amniotic fluid levels of annexin A2 in patients with preterm premature rupture of the membranes was significantly higher than that in the remainder (P=0.002). Amniotic levels of annexin A2 was a fair test (area under receiver-operator characteristic curve=0.679), and amniotic fluid levels of annexin A2>878.2ng/mL had a sensitivity of 68.8%, a specificity of 65.2%, a positive predictive value of 73.3%, and a negative predictive value of 60.0% for predicting delivery within 2weeks after amniotic fluid sampling. Furthermore, the combined use of amniotic fluid cut-off levels of 878.2ng/mL for annexin A2 and 13.3ng/mL for interleukin-8 improved the specificity (91.3%) and the positive predictive value (89.5%). We identified amniotic fluid levels of annexin A2, especially in combination with amniotic fluid levels of interleukin-8, as a novel predictive marker for preterm delivery. © 2011 The Authors. Journal of Obstetrics and Gynaecology Research © 2011 Japan Society of Obstetrics and Gynecology.

  16. Clinical observation of corneal lamellar debridement combined with sutureless amniotic membrane transplantation for the treatment of superficial fungal keratitis

    Directory of Open Access Journals (Sweden)

    Huang Zhang

    2014-09-01

    Full Text Available AIM:To evaluate the clinical efficacy of corneal lamellar debridement combined with sutureless amniotic membrane transplantation for the treatment of superficial fungal keratitis.METHODS:Totally 22 cases(22 eyeswith superficial fungal keratitis were referred to our hospital from April 2012 to October 2013. The patients with persistent cornea ulcer after treatment of local and systemic antifungal drugs underwent corneal lamellar debridement combined with sutureless amniotic membrane transplantation, and the recipient bed was covered with an amniotic membrane using fibrin sealant during the operation. All patients were still given topical antifungal therapy for 1-2mo after operation. The followed-up time was 3mo or above. We observed the corneal healing and amniotic membrane adhesion by split lamp microscope, and investigated the transformation of amniotic membrane and fungal infection recurrence with confocal microscope. RESULTS: Corneal edema and anterior chamber reaction of 21 patients disappeared gradually, and no amniotic membrane graft dissolved and shed off within 1-2wk postoperatively. Two weeks after operation, the graft integrated into the corneal and the corneal wounds' thickness increased gradually, the corneal epithelium reconstructed and corneas became clear. Four weeks after operation, the corneal scarring developed gradually and fluorescence staining was negative. Nineteen cases' amniotic membranes that adhered with the cornea dissolved 4wk after operation. There were different degrees of corneal nebula or macula remained 3mo postoperatively. All patients' vision improved in varying degrees, except in 1 case with fungal keratitis who had been cured by lamellar keratoplasty.CONCLUSION:Corneal lamellar debridement combined with sutureless amniotic membrane transplantation can effectively remove the foci of inflammation, improve the local efficacy, shorten the operation time, relieve the postoperative reaction, and promote cornea

  17. Periodontal disease and intra-amniotic complications in women with preterm prelabor rupture of membranes.

    Science.gov (United States)

    Radochova, Vladimira; Kacerovska Musilova, Ivana; Stepan, Martin; Vescicik, Peter; Slezak, Radovan; Jacobsson, Bo; Kacerovsky, Marian

    2017-08-04

    Periodontal disease is frequently suggested as a possible causal factor for preterm delivery. The link between periodontal disease and preterm delivery is a possible translocation of periopathogenic bacteria to the placenta and amniotic fluid as well as a systemic response to this chronic inflammatory disease. However, there is a lack of information on whether there is an association between clinical periodontal status in women with preterm prelabor rupture of membranes (PPROM) and the presence of microbial invasion of the amniotic cavity (MIAC) and intra-amniotic inflammation (IAI). Therefore, the main aim of this study was to evaluate the incidence and severity of periodontal disease in women with PPROM. The secondary aim was to characterize an association between periodontal status and the presence of intra-amniotic PPROM complications (MIAC and/or IAI). Seventy-eight women with PPROM at gestational ages between 24 + 0 and 36 + 6 weeks were included in this study. The samples of amniotic fluid were obtained at admission via transabdominal amniocentesis, and amniotic fluid interleukin (IL)-6 concentrations were determined using a point-of-care test. All women had a full-mouth recording to determine the periodontal and oral hygiene status. Probing pocket depth and clinical attachment loss were measured at four sites on each fully erupted tooth. In total, 45% (35/78) of women with PPROM had periodontal disease. Mild, moderate, and severe periodontal disease was present in 19% (15/78), 19% (15/78), and 6% (5/78) of women, respectively. The presence of MIAC and IAI was found in 28% (22/78) and 26% (20/78) of women, respectively. Periopathogenic bacteria (2 × Streptococcus intermedius and 1 × Fusobacterium nucleatum) was found in the amniotic fluid of 4% (3/78) of women. There were no differences in periodontal status between women with MIAC and/or IAI and women without these intra-amniotic complications. The presence of MIAC and IAI was not related

  18. Impact of application of bio-amniotic membrane immersed in 5-fluorouracil solution in trabeculectomy on rabbit retina

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    Chenming Zhang

    2013-01-01

    Full Text Available Background : To observe the impact of application of bio-amniotic membrane immersed in 5-fluorouracil solution in trabeculectomy on the retina in a rabbit model. Materials and Methods : Healthy white New Zealand rabbits were randomly assigned into three groups with 20 in each group. Bio-amniotic membranes of 4 × 5 mm immersed in either physiological saline/water for 10 min, or 25 mg/mL 5-fluorouracil solution for 5 and 10 min, respectively, were applied on rabbit eyes during trabeculectomy. At 7, 14, 21, and 28 days of postoperation, five rabbits from each group were examined with electroretinogram (ERG. After being examined for eye pressure and bleb morphology, rabbits were sacrificed by air embolism and their retinas were collected and examined by transmission electron microscopy (TEM. In addition, 5-fluorouracil amount in bio-amniotic membranes was measured using high-performance liquid chromatography. Results: Each bio-amniotic membrane could absorb 59.004 μg and 75.828 μg 5-fluorouracil after being immersed in 5-fluorouracil solution for 5 and 10 min, respectively. Application of these bio-amniotic membranes in trabeculectomy could promote the formation of well-functioning bleb and maintain intraocular pressure, although it had no effect on retina structures as examined with ERG and TEM. Conclusion: Application of 5-FU soaked bio-amniotic membrane in rabbit eye trabeculectomy is effective and safe.

  19. Application of Amniotic Membrane in Ocular Surface Diseases: Clinical Features and Treatment Outcome

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    Derya Cindarik

    2012-05-01

    Full Text Available Pur po se: To investigate the effectiveness of amniotic membrane transplantation in cases with corneal thinning, desmatocele and refractive corneal ulcer. Ma te ri al and Met hod: Fifty-four eyes of 54 patients who were applied amniotic membrane transplantation for various ocular surface disease between January 2004 and February 2009 in Çukurova University Ophthalmology Department were included in the study. A complete ophthalmologic examination was performed. Corneal culture and corneal cytology samples were collected from the patients with the diagnosis of corneal ulcers. The patients were informed about the surgical procedure and the possible complications and informed consent was obtained. The amniotic membranes that were prepared under optimal conditions and protected in frozen forms were used in the operations. Follow-up examinations were done at postoperative 1st day, 1st week, 1st month, 3rd month, 6th month and then once in a year. Re sults: Of 54 patients, 26 (48.1% were men and 28 (51.8% were women. The mean age of patients was 52.53±19.75 (2-87 years. The cases were separated into 2 groups according to the etiology: group 1 - eyes with corneal ulcer (n:26 and group 2 - eyes with corneal stromal thinning, persistent epithelial defects and desmatocel (n:28. The transplantations were performed using cover technique in 17 eyes (31.4%, graft technique in 37 eyes (68.5% and graft technique with corneal patch in 2 eyes (3.7%. Partial penetrating keratoplasty was required in 38 of 54 eyes (70.3%. One eye was enucleated. Dis cus si on: The amniotic membrane transplantation has advantages like: it can be prepared easily and is cost-effective. It is a safe and effective procedure in ocular surface disease. (Turk J Ophthalmol 2012; 42: 177-82

  20. In vivo studies of antibacterial effect of human amniotic membrane use in treatment of burns

    International Nuclear Information System (INIS)

    Zobiri, A.; Moussi, W.; Djeffal, A.; Larbi Daho Bachir, M.

    2001-01-01

    The present study consist to put in evidence one of essential characteristic of amniotic membrane in occurrence the antibacterial effect. The article describes a study which compared the microbiological and clinical results of the application of freeze-dried gamma sterilized amniotic membrane with that of the conventional treatment(flamazine, greased gauze), in 100 patients with intermediate burns and deep burns with small surface, the bacterial population of various types of microorganisms was well controlled using quantitative bacterial culture techniques, immediately after accident, during treatment and at the last of treatment. The bacterial counts were significantly diminished mean of 10 4 UFC/cm2 to mean of 10 3 UFC/cm2, after 4 days in no infected burns and mean of 10 6 UFC/cm2 to mean of 10 3 UFC/cm2, after 5 days in infected burns, the same reduction was registered after 16 to 25 days with the conventional treatment. It is concluded that use of amniotic membrane control infection, minimize pain and promote wound healing

  1. Effectiveness of the radio sterilized amniotic membrane transplantation vs conjunctival auto graft implant in the pterygium surgery with intraoperative mitomycin C

    International Nuclear Information System (INIS)

    Tellez Y, L.; Martinez P, M. E.; Vazquez M, L.

    2009-01-01

    At the present time the traditional surgical handling of the pterygium with conjunctival implant presents high frequency. In this work the obtained results of a controlled clinic practice blind double are presented of the period of December from 2008 to December 2009, realized in the Medical Unit of Ambulatory Attention 231 of the Mexican Institute of the Public Health located in Metepec, Mexico State, in order to demonstrate the effectiveness and security of the alternative use of the radio sterilized amniotic membrane and intraoperative mitomycin C. For the study was included patients with diagnostic of primary nasal pterygium in one or both eyes, adults of both genus with an age range of 20 to 60 years, and was used radio sterilized amniotic membrane processed in the Bank of Radio Sterilized Tissues of the National Institute of Nuclear Research. (Author)

  2. Effects of ionizing radiation on glycerolated amniotic membranes as a substract for cultured human epithelium

    International Nuclear Information System (INIS)

    Paggiaro, Andre Oliveira

    2011-01-01

    The amniotic membrane (AM) is a biomaterial with biological properties that are beneficial to tissue repair. It has been used as a temporary coverage to threat burns and chronic wounds. Recently, it has been served as a substrate for keratinocytes culture to construct a living skin equivalent. However, MA is a biological material, and its transplantation could cause infectious disease for receptors. So, it must be preserved and sterilized before clinical use. The aim of this study was to evaluate the radiation effects on glycerol-preserved MA, considering its compatibility to support human keratinocytes culture. Four MA were stored in high concentrations of glycerol (> 85%) and half of them were radio sterilized with a dose of 25 kGy. Then, we established two groups: nonirradiated MA (MA-ni) and irradiated MA (MA-i). Both groups was deepithelialized by a standardized protocol and was investigated morphologically, immunohistochemical and ultrastructural. Subsequently human keratinocytes were cultivated immersed and in air-liquid interface on denuded surface of MA-i and MA-ni. The results were compared at 14 and 21 days of culture by light and electron microscopy. After epithelial denudation, analyses demonstrated the continuity of the basement membrane in MA-ni group, whereas in the irradiated group, there was no indication of the basement membrane’s presence on the surface of MA. The cell cultures showed that in the non-irradiated group, there was growth of a multi-layered and differentiated epithelium, with a stratum corneum’s formation in air-liquid interface. In the irradiated group, the epithelium had only two or three layer, little cell differentiation, with the same results immersed or air-liquid interface system. Glycerol-preserved MA was biocompatible with the growth of a cultivated epithelium, showing its potential as a skin substitute. Irradiation at 25 kGy cause structural damage to the tissue, making changes in basement membrane, that facilitates

  3. Stabilization of collagen nanofibers with l-lysine improves the ability of carbodiimide cross-linked amniotic membranes to preserve limbal epithelial progenitor cells

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    Lai JY

    2014-11-01

    Full Text Available Jui-Yang Lai,1–3 Pei-Ran Wang,1 Li-Jyuan Luo,1 Si-Tan Chen1 1Institute of Biochemical and Biomedical Engineering, 2Biomedical Engineering Research Center, 3Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan, Republic of ChinaAbstract: To overcome the drawbacks associated with limited cross-linking efficiency of carbodiimide modified amniotic membrane, this study investigated the use of L-lysine as an additional amino acid bridge to enhance the stability of a nanofibrous tissue matrix for a limbal epithelial cell culture platform. Results of ninhydrin assays and zeta potential measurements showed that the amount of positively charged amino acid residues incorporated into the tissue collagen chains is highly correlated with the L-lysine -pretreated concentration. The cross-linked structure and hydrophilicity of amniotic membrane scaffolding materials affected by the lysine molecular bridging effects were determined. With an increase in the L-lysine-pretreated concentration from 1 to 30 mM, the cross-linking density was significantly increased and water content was markedly decreased. The variations in resistance to thermal denaturation and enzymatic degradation were in accordance with the number of cross-links per unit mass of amniotic membrane, indicating L-lysine-modulated stabilization of collagen molecules. It was also noteworthy that the carbodiimide cross-linked tissue samples prepared using a relatively high L-lysine-pretreated concentration (ie, 30 mM appeared to have decreased light transmittance and biocompatibility, probably due to the influence of a large nanofiber size and a high charge density. The rise in stemness gene and protein expression levels was dependent on improved cross-link formation, suggesting the crucial role of amino acid bridges in constructing suitable scaffolds to preserve limbal progenitor cells. It is concluded that mild to moderate pretreatment conditions (ie, 3–10 mM L-lysine can

  4. Antioxidant Vitamin Status in the Serum and Amniotic Fluid of Women with Premature Rupture of the Fetal Membranes.

    Science.gov (United States)

    Barrett, Bridget M.

    The purpose of this study was to examine the status of antioxidant vitamins in women with premature rupture of the fetal membranes. Specimens of blood and amniotic fluid were obtained from 80 pregnant subjects included both smokers and non-smokers during the third trimester. The concentrations of ascorbic acid (ASA), beta -carotene, retinol and alpha -tocopherol in serum and amniotic fluid were determined. The experimental group consisted of those subjects with PROM while the control subjects were those with normal pregnancy. No statistical differences were found between the PROM and control groups in retinol and vitamin E concentrations in amniotic fluid and serum. Serum ASA concentrations of PROM subjects were not different from controls, but the PROM subjects had significantly lower amniotic fluid ASA concentrations. However, in a study with fewer subjects a lower serum ASA concentration in the PROM subjects was observed. The ratio of amniotic fluid ASA concentration to ASA serum concentration was significantly lower in PROM patients than in controls in both studies. This suggests that low levels of ASA in the amniotic fluid, but not in serum is better associated with PROM. A low amniotic fluid concentration of ASA may reflect an inefficient transfer and/or increased fetal utilization. Alterations in ASA concentration in the amniotic fluid may affect the integrity of the chorioamnion leading to PROM. beta -Carotene was not found in the amniotic fluid. Serum beta-carotene levels were significantly lower in the PROM group compared to the control group. Low concentrations of beta-carotene in maternal serum in smokers not only associated with poor maternal outcome (PROM) but also compromised the fetal outcome (decreased birth weight). Maintenance of adequate serum beta-carotene concentration and amniotic fluid ASA in smokers may result in better maternal and fetal outcome. This study demonstrated that nutrition is an important factor in the prevention of PROM.

  5. Corneal regeneration by induced human buccal mucosa cultivated on an amniotic membrane following alkaline injury.

    Science.gov (United States)

    Man, Rohaina Che; Yong, Then Kong; Hwei, Ng Min; Halim, Wan Haslina Wan Abdul; Zahidin, Aida Zairani Mohd; Ramli, Roszalina; Saim, Aminuddin Bin; Idrus, Ruszymah Binti Hj

    2017-01-01

    Various clinical disorders and injuries, such as chemical, thermal, or mechanical injuries, may lead to corneal loss that results in blindness. PURPOSE : The aims of this study were to differentiate human buccal mucosa (BMuc) into corneal epithelial-like cells, to fabricate engineered corneal tissue using buccal mucosal epithelial cells, and to reconstruct a damaged corneal epithelium in a nude rat model. BMuc were subjected to 10 d of induction factors to investigate the potential of cells to differentiate into corneal lineages. Corneal stem cell markers β1-integrin, C/EBPδ, ABCG2, p63, and CK3 were upregulated in the gene expression analysis in induced BMuc, whereas CK3 and p63 showed significant protein expression in induced BMuc compared to the uninduced cells. BMuc were then left to reach 80% confluency after differential trypsinization. The cells were harvested and cultivated on a commercially available untreated air-dried amniotic membrane (AM) in a Transwell system in induction medium. The corneal constructs were fabricated and then implanted into damaged rat corneas for up to 8 weeks. A significant improvement was detected in the treatment group at 8 weeks post-implantation, as revealed by slit lamp biomicroscopy analysis. The structure and thickness of the corneal layer were also analyzed using histological staining and time-domain optical coherence tomography scans and were found to resemble a native corneal layer. The protein expression for CK3 and p63 were continuously detected throughout the corneal epithelial layer in the corneal construct. In conclusion, human BMuc can be induced to express a corneal epithelial-like phenotype. The addition of BMuc improves corneal clarity, prevents vascularization, increases corneal thickness and stromal alignment, and appears to have no adverse effect on the host after implantation.

  6. Novel approach to gastric mucosal defect repair using fresh amniotic membrane allograft in dogs (experimental study).

    Science.gov (United States)

    Farghali, Haithem A; AbdElKader, Naglaa A; Khattab, Marwa S; AbuBakr, Huda O

    2017-10-18

    Gastric mucosal defect could result from several causative factors including the use of nonsteroidal anti-inflammatory drugs, Helicobacter pylori infection, gastrointestinal and spinal cord diseases, and neoplasia. This study was performed to achieve a novel simple, inexpensive, and effective surgical technique for the repair of gastric mucosal defect. Six adult male mongrel dogs were divided into two groups (three dogs each). In the control positive group (C + ve), dogs were subjected to surgical induction of gastric mucosal defect and then treated using traditional medicinal treatment for such a condition. In the amniotic membrane (AM) group, dogs were subjected to the same operation and then fresh AM allograft was applied. Clinical, endoscopic, biochemical (serum protein and lipid and pepsin activity in gastric juice), histopathological, and immunohistochemistry evaluations were performed. Regarding endoscopic examination, there was no sign of inflammatory reaction around the grafted area in the AM group compared to the C + ve group. The leukocytic infiltration in the gastric ulcer was well detected in the control group and was less observed in the AM group. In the AM group, the concentrations of both protein and lipid profiles were nearly the same as those in serum samples taken preoperatively at zero time, which indicated that the AM grafting acted the same as gastric mucosa. The re-epithelization of the gastric ulcer in the C + ve group was not yet detected at 21 days, while in the AM group it was well observed covering most of the gastric ulcer. AM accelerated the re-epithelization of the gastric ulcer. The fibrous connective tissue and the precursor of collagen (COL IA1) were poorly detected in the gastric ulcer with AM application. Using fresh AM allograft for repairing gastric mucosal defect in dogs showed great impact as a novel method to achieve optimum reconstruction of the gastric mucosal architecture and restoration of pre

  7. Amnioinfusion in term labor with low amniotic fluid due to rupture of membranes: a new indication.

    Science.gov (United States)

    Miño, M; Puertas, A; Miranda, J A; Herruzo, A J

    1999-01-01

    The null hypothesis was that the use of intrapartum amnioinfusion to induce term labor because of premature rupture of membranes when labor was complicated by low amniotic fluid volume due to vaginal loss would not improve fetal heart rate patterns, decrease the incidence of operative delivery, or improve neonatal acid-base status. 200 term pregnancies with low amniotic fluid due to vaginal loss were randomly chosen to receive intrapartum amnioinfusion or standard obstetric care without amnioinfusion. Fetal heart rate pattern, method of delivery and neonatal acid-base status were compared with Student's t test, chi-squared analysis, Mann-Whitney U- or Fisher's exact test. When amnioinfusion was used, the fetuses had lower rates of variable (74 vs. 91%, Pamnioinfusion, and babies in this group had lower rates of neonatal acidemia of arterial (22 vs. 36%, PAmnioinfusion improved fetal heart rate pattern, lowered the incidence of operative delivery, and improved neonatal acid-base status in term labor complicated by low amniotic fluid due to vaginal loss.

  8. Basic study of healing of injuries to the myometrium and amniotic membrane using photocrosslinkable chitosan.

    Science.gov (United States)

    Suzuki, Keitaro; Shinya, Masaru; Kitagawa, Michihiro

    2006-04-01

    We performed in vivo and in vitro adhesion experiments on the use of photocrosslinkable chitosan (PC) for healing of the myometrium and the amniotic membrane to test its usefulness in the perinatal field. To determine the adhesion conditions of PC in the healing of the myometrium and amniotic membrane, we applied two concentrations (30 and 50 mg/mL) of PC to the postpartum amniotic membrane, and radiated it with ultraviolet (UV) light for 10, 20, 30 or 60 s. Next, we created incisions of three sizes (pinhole, 5 mm and 10 mm), and performed PC adhesion followed by pressure application experiments. Subsequently, we laparotomized 12-day-pregnant rats under anesthesia, created a 5-mm incision, applied PC and closed the abdomen. We removed the uterus at 20 days of pregnancy, and histologically examined the site of PC application. The optimal adhesion conditions of PC were 50 mg/mL, and the optimal time of UV irradiation was 30 s or longer. In pressure application experiments, the bonding strength of PC was so high that even 10-mm incisions withstood a mean pressure of 92 mmH2O. The bonding strength of PC was significantly higher than that of fibrin glue that was tested in parallel. Histological examination of the rat myometrium showed a mild foreign body reaction with no inflammatory findings, confirming the usefulness of PC. PC is a wound-healing drug, an improvement on the naturally occurring polysaccharide chitosan, and is safer and has a higher bonding strength than biological adhesives including fibrin, making it promising for fetal surgical and perinatal applications.

  9. [Successful treatment of surgically induced necrotizing sclerokeratitis (SINS) with systemic immunosuppresive agents and amniotic membrane grafting].

    Science.gov (United States)

    Cordero-Coma, M; Franco-Benito, M; García-Ruiz-de-Morales, J M; Alonso-Orcajo, N; Del Barrio-Manso, I

    2009-11-01

    We report the case of a 74-year-old female who developed a necrotizing sclerokeratitis affecting her left eye after uncomplicated cataract surgery. She had no previous history of systemic autoimmune disease. Histopathology of the lesion revealed necrotic granulomatosis with an increased number of plasma cells. Surgically induced necrotizing sclerokeratitis (SINS) is a serious entity which requires prompt and aggressive therapy to prevent its potential devastating ocular consequences. Conjunctival resection and amniotic membrane grafting may be necessary to temporarily interrupt local immunologic events in severe cases. However, associated systemic immunomodulatory therapy seems to be mandatory (Arch Soc Esp Oftalmol 2009; 84: 577-580).

  10. Anti-aging Effect of Transplanted Amniotic Membrane Mesenchymal Stem Cells in a Premature Aging Model of Bmi-1 Deficiency

    Science.gov (United States)

    Xie, Chunfeng; Jin, Jianliang; Lv, Xianhui; Tao, Jianguo; Wang, Rong; Miao, Dengshun

    2015-01-01

    To determine whether transplanted amniotic membrane mesenchymal stem cells (AMSCs) ameliorated the premature senescent phenotype of Bmi-1-deficient mice, postnatal 2-day-old Bmi-1−/− mice were injected intraperitoneally with the second-passage AMSCs from amniotic membranes of β-galactosidase (β-gal) transgenic mice or wild-type (WT) mice labeled with DiI. Three reinjections were given, once every seven days. Phenotypes of 5-week-old β-gal+ AMSC-transplanted or 6-week-old DiI+ AMSC-transplanted Bmi-1−/− mice were compared with vehicle-transplanted Bmi-1−/− and WT mice. Vehicle-transplanted Bmi-1−/− mice displayed growth retardation and premature aging with decreased cell proliferation and increased cell apoptosis; a decreased ratio and dysmaturity of lymphocytic series; premature osteoporosis with reduced osteogenesis and increased adipogenesis; redox imbalance and DNA damage in multiple organs. Transplanted AMSCs carried Bmi-1 migrated into multiple organs, proliferated and differentiated into multiple tissue cells, promoted growth and delayed senescence in Bmi-1−/− transplant recipients. The dysmaturity of lymphocytic series were ameliorated, premature osteoporosis were rescued by promoting osteogenesis and inhibiting adipogenesis, the oxidative stress and DNA damage in multiple organs were inhibited by the AMSC transplantation in Bmi-1−/− mice. These findings indicate that AMSC transplantation ameliorated the premature senescent phenotype of Bmi-1-deficient mice and could be a novel therapy to delay aging and prevent aging-associated degenerative diseases. PMID:26370922

  11. Co-effect of silk and amniotic membrane for tendon repair.

    Science.gov (United States)

    Seo, Young-Kwon; Kim, Jun-Hyung; Eo, Su-Rak

    2016-08-01

    The objective of the present study was to determine the feasibility and biocompatibility of a silk scaffold and a composite silk scaffold in terms of new tendon generation using a rabbit Achilles tendon model. The silk scaffold was constructed using a weaving machine, then soaked in a 1% collagen-hyaluronan (HA) solution and air-dried, whereas the composite silk scaffold was composed of a silk scaffold containing a lyophilized collagen-HA substrate. Tenocytes were cultured in vitro to compare cell populations in the two groups. The cellular densities on composite silk scaffolds were 40% higher on average than those on silk scaffolds in 30-day tenocyte cultures. The tendon scaffolds had implanted into Achilles tendon defects in 16 white New Zealand rabbits. Rabbits were randomly divided into the following three groups: group I, silk scaffold alone; group II, composite silk scaffold; and group III, composite silk scaffold wrapped by an amniotic membrane. Implants were harvested 2, 8, and 12 weeks post-implantation. Histological examinations were conducted using hematoxylin-eosin (H&E), Masson's trichrome, and by performing immunohistochemical staining for CD34. After 12 weeks, the three groups were distinguishable based on gross examination. The histological examination revealed more organized collagen fibrils in groups III, which showed a dense, parallel, linear organization of collagen bundles. CD34 staining revealed neoangiogenesis in groups III. The results of this research showed that collagen-HA substrates with amniotic membrane accelerate cellular migration and angiogenesis in neotendons.

  12. Amniotic Membrane Transplant with a Special Technique (Motowa's Sandwich Technique) in Mooren's Ulcer.

    Science.gov (United States)

    Al Motowa, Saeed; Al Zobidi, Mohammed

    2015-01-01

    To illustrate amniotic membrane transplant (AMT) with a novel surgical technique ("sandwich technique") for treating patients with malignant Mooren's ulcer. A case report of a patient with bilateral, malignant Mooren's ulcer who had undergone systemic steroid therapy and topical immunosuppresive therapy to stabilize his condition. However, perforation of cornea occurred in one eye. AMT with a new surgical technique ("Motowa's sandwich technique") was performed to treat this case. On the 1(st) day postoperatively, there was no pain, no photophobia, and visual acuity was same as preoperatively. At 4 weeks postoperatively, visual acuity improved in the right eye to 20/160 with pinhole, and there was no fluorescein staining. The right eye was quiet. Amniotic membrane transplant with "Motowa's sandwich technique" resulted in the preservation of the anatomical integrity and progression of disease was halted along with an improved vision. This technique is a novel surgical modality in treating Mooren's ulcer. Further study on a large cohort of patients is required for evidence-based data to verify the outcome of this initial case report.

  13. Clinical outcome of combined conjunctival autograft transplantation and amniotic membrane transplantation in pterygium surgery

    Science.gov (United States)

    Malla, Tejsu; Jiang, Jing; Hu, Kai

    2018-01-01

    AIM To compare long-term outcome of primary and recurrent pterygium surgery with three different techniques: combined conjunctival autograft and overlay amniotic membrane transplantation (CAT with AMT), conjunctival autograft transplantation (CAT) alone and amniotic membrane transplantation (AMT) alone. METHODS In this retrospective study, 142 eyes of 142 pterygium patients (104 primary, 38 recurrent) who underwent CAT (group A), AMT (group B) or CAT with AMT (group C) respectively following surgical excision were reviewed and compared based on the recurrences and post-operative complications. RESULTS The number of recurrence post-surgery were 17 (9 from primary, 8 from recurrent; the same description below), 18 (10, 8) and 2 (1, 1) in groups A, B, and C respectively; dry eyes were 22 (16, 6), 27 (18, 9) and 7 (3, 4); conjunctival inflammations were 30 (17, 13), 27 (16, 11) and 11 (6, 5). Patients in group C (either primary or recurrent or both) mainly showed significantly better results than those in group A or B (P<0.05) regarding above-mentioned clinical effects. CONCLUSION Combined CAT and overly AMT have significantly lower rates of recurrence and postoperative complications for primary and recurrent pterygium surgery than CAT or AMT alone. PMID:29600172

  14. Clinical outcome of combined conjunctival autograft transplantation and amniotic membrane transplantation in pterygium surgery

    Directory of Open Access Journals (Sweden)

    Tejsu Malla

    2018-03-01

    Full Text Available AIM: To compare long-term outcome of primary and recurrent pterygium surgery with three different techniques: combined conjunctival autograft and overlay amniotic membrane transplantation (CAT with AMT, conjunctival autograft transplantation (CAT alone and amniotic membrane transplantation (AMT alone. METHODS: In this retrospective study, 142 eyes of 142 pterygium patients (104 primary, 38 recurrent who underwent CAT (group A, AMT (group B or CAT with AMT (group C respectively following surgical excision were reviewed and compared based on the recurrences and post-operative complications. RESULTS: The number of recurrence post-surgery were 17 (9 from primary, 8 from recurrent; the same description below, 18 (10, 8 and 2 (1, 1 in groups A, B, and C respectively; dry eyes were 22 (16, 6, 27 (18, 9 and 7 (3, 4; conjunctival inflammations were 30 (17, 13, 27 (16, 11 and 11 (6, 5. Patients in group C (either primary or recurrent or both mainly showed significantly better results than those in group A or B (P<0.05 regarding above-mentioned clinical effects. CONCLUSION: Combined CAT and overly AMT have significantly lower rates of recurrence and postoperative complications for primary and recurrent pterygium surgery than CAT or AMT alone.

  15. Use of Extracorporeal Membrane Oxygenation in a Fulminant Course of Amniotic Fluid Embolism Syndrome Immediately after Cesarean Delivery

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    Jae Ha Lee

    2016-08-01

    Full Text Available Amniotic fluid embolism is rare but is one of the most catastrophic complications in the peripartum period. This syndrome is caused by a maternal anaphylactic reaction to the introduction of fetal material into the pulmonary circulation. When amniotic fluid embolism is suspected, the immediate application of extracorporeal mechanical circulatory support such as veno-arterial extracorporeal membrane oxygenation (ECMO or cardiopulmonary bypass should be considered. Without the application of extracorporeal mechanical circulatory support, medical supportive care might not be sufficient to maintain cardiopulmonary stabilization in severe cases of amniotic fluid embolism. In this report, we present the case of a 36-year-old pregnant woman who developed an amniotic fluid embolism immediately after a cesarean section. Her catastrophic event started with the sudden onset of severe hypoxia, followed by circulatory collapse within 8 minutes. The veno-arterial mode of extracorporeal membrane oxygenation was initiated immediately. She was successfully resuscitated but with impaired cognitive function. Thus, urgent ECMO should be considered when amniotic fluid embolism syndrome is suspected in patients presenting acute cardiopulmonary collapse.

  16. Amniotic membrane-derived stem cells: immunomodulatory properties and potential clinical application

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    Insausti CL

    2014-03-01

    Full Text Available Carmen L Insausti,1 Miguel Blanquer,1 Ana M García-Hernández,1 Gregorio Castellanos,2 José M Moraleda11Unidad de Trasplante Hematopoyético y Terapia Celular, 2Servicio de Cirugía, Hospital Clínico Universitario Virgen de la Arrixaca, IMIB, Campus Mare Nostrum, Universidad de Murcia, El Palmar, Murcia, SpainAbstract: Epithelial and mesenchymal cells isolated from the amniotic membrane (AM possess stem cell characteristics, differentiation potential toward lineages of different germ layers, and immunomodulatory properties. While their expansion and differentiation potential have been well studied and characterized, knowledge about their immunomodulatory properties and the mechanisms involved is still incomplete. These mechanisms have been evaluated on various target cells of the innate and the adaptive system and in animal models of different inflammatory diseases. Some results have evidenced that the immunomodulatory effect of AM-derived cells is dependent on cell-cell contact, but many of them have demonstrated that these properties are mediated through the secretion of suppressive molecules. In this review, we present an update on the described immunomodulatory properties of the derived amniotic cells and some of the proposed involved mechanisms. Furthermore, we describe some assays in animal models of different inflammatory diseases which reveal the potential use of these cells to treat such diseases.Keywords: epithelial cells, mesenchymal cells, cell therapy, immunomodulation

  17. A clinical trial on the treatment of burns in children at the University Teaching Hospital using amniotic membrane sterilized by gamma radiation (July to December 1994)

    International Nuclear Information System (INIS)

    Katebe, K.R.C.

    1995-01-01

    About six hundred patients are admitted to the University Teaching Hospital every year. Most of the burns are from water and fire with a high mortality rate in children. This study was centred on the feasibility of using amniotic membrane for wound dressing. The method of Gamma radiation was employed to sterilize this biological tissue before utilising it on patients. The results of the study show more superior healing rate compared with other methods. There was lack of hypersensitivity reactions and reduced wound infection and good quality wound healing. The method is easy to use and has been accepted by the patients and nursing staff. (n.m.). 20 refs., tabs., 6 graphs

  18. Glutaraldehyde cross-linking of amniotic membranes affects their nanofibrous structures and limbal epithelial cell culture characteristics

    Directory of Open Access Journals (Sweden)

    Lai JY

    2013-10-01

    Full Text Available Jui-Yang Lai,1–3 David Hui-Kang Ma4,5 1Institute of Biochemical and Biomedical Engineering, 2Biomedical Engineering Research Center, 3Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; 4Limbal Stem Cell Laboratory, Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan; 5Department of Chinese Medicine, Chang Gung University, Taoyuan, Taiwan Abstract: Given that the cells can sense nanometer dimensions, the chemical cross-linking-mediated alteration in fibrillar structure of collagenous tissue scaffolds is critical to determining their cell culture performances. This article explores, for the first time, the effect of nanofibrous structure of glutaraldehyde (GTA cross-linked amniotic membrane (AM on limbal epithelial cell (LEC cultivation. Results of ninhydrin assays demonstrated that the amount of new cross-links formed between the collagen chains is significantly increased with increasing the cross-linking time from 1 to 24 hours. By transmission electron microscopy, the AM treated with GTA for a longer duration exhibited a greater extent of molecular aggregation, thereby leading to a considerable increase in nanofiber diameter and resistance against collagenase degradation. In vitro biocompatibility studies showed that the samples cross-linked with GTA for 24 hours are not well-tolerated by the human corneal epithelial cell cultures. When the treatment duration is less than 6 hours, the biological tissues cross-linked with GTA for a longer time may cause slight reductions in 3-(4,5-dimethylthiazol-2-yl-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl-2H-tetrazolium, inner salt, and anti-inflammatory activities. Nevertheless, significant collagen molecular aggregation also enhances the stemness gene expression, indicating a high ability of these AM matrices to preserve the progenitors of LECs in vitro. It is concluded that GTA cross-linking of collagenous tissue materials may affect their nanofibrous

  19. Combined Conjunctival Autograft and Overlay Amniotic Membrane Transplantation; a Novel Surgical Treatment for Pterygium

    Directory of Open Access Journals (Sweden)

    Siamak Zarei Ghanavati

    2014-01-01

    Full Text Available The authors report the long-term results of combined conjunctival autograft and overlay amniotic membrane transplantation (AMT for treatment of pterygium as a new surgical technique. Nineteen patients including 12 male and 7 female subjects with pterygium (primary, 14 cases; recurrent, 5 cases underwent combined conjunctival autograft and overlay AMT and were followed from 10 to 26 months. Mean age was 44.21±12.49 (range, 29.0-73.0 years. In one patient with grade T3 primary pterygium, the lesion recurred (5.2%, recurrence rate. No intra-and postoperative complication developed. This procedure seems a safe and effective surgical technique for pterygium treatment. Protection of the ocular surface during the early postoperative period reduces the friction-induced inflammation and might be helpful to prevent the recurrence.

  20. Amniotic membrane grafts for the prevention of esophageal stricture after circumferential endoscopic submucosal dissection.

    Science.gov (United States)

    Barret, Maximilien; Pratico, Carlos Alberto; Camus, Marine; Beuvon, Frédéric; Jarraya, Mohamed; Nicco, Carole; Mangialavori, Luigi; Chaussade, Stanislas; Batteux, Frédéric; Prat, Frédéric

    2014-01-01

    The prevention of esophageal strictures following circumferential mucosal resection remains a major clinical challenge. Human amniotic membrane (AM) is an easily available material, which is widely used in ophthalmology due to its wound healing, anti-inflammatory and anti-fibrotic properties. We studied the effect of AM grafts in the prevention of esophageal stricture after endoscopic submucosal dissection (ESD) in a swine model. In this prospective, randomized controlled trial, 20 swine underwent a 5 cm-long circumferential ESD of the lower esophagus. In the AM Group (n = 10), amniotic membrane grafts were placed on esophageal stents; a subgroup of 5 swine (AM 1 group) was sacrificed on day 14, whereas the other 5 animals (AM 2 group) were kept alive. The esophageal stent (ES) group (n = 5) had ES placement alone after ESD. Another 5 animals served as a control group with only ESD. The prevalence of symptomatic strictures at day 14 was significantly reduced in the AM group and ES groups vs. the control group (33%, 40% and 100%, respectively, p = 0.03); mean esophageal diameter was 5.8±3.6 mm, 6.8±3.3 mm, and 2.6±1.7 mm for AM, ES, and control groups, respectively. Median (range) esophageal fibrosis thickness was 0.87 mm (0.78-1.72), 1.19 mm (0.28-1.95), and 1.65 mm (0.7-1.79) for AM 1, ES, and control groups, respectively. All animals had developed esophageal strictures by day 35. The anti-fibrotic effect of AM on esophageal wound healing after ESD delayed the development of esophageal stricture in our model. However, this benefit was of limited duration in the conditions of our study.

  1. Amniotic membrane grafts for the prevention of esophageal stricture after circumferential endoscopic submucosal dissection.

    Directory of Open Access Journals (Sweden)

    Maximilien Barret

    Full Text Available The prevention of esophageal strictures following circumferential mucosal resection remains a major clinical challenge. Human amniotic membrane (AM is an easily available material, which is widely used in ophthalmology due to its wound healing, anti-inflammatory and anti-fibrotic properties. We studied the effect of AM grafts in the prevention of esophageal stricture after endoscopic submucosal dissection (ESD in a swine model.In this prospective, randomized controlled trial, 20 swine underwent a 5 cm-long circumferential ESD of the lower esophagus. In the AM Group (n = 10, amniotic membrane grafts were placed on esophageal stents; a subgroup of 5 swine (AM 1 group was sacrificed on day 14, whereas the other 5 animals (AM 2 group were kept alive. The esophageal stent (ES group (n = 5 had ES placement alone after ESD. Another 5 animals served as a control group with only ESD.The prevalence of symptomatic strictures at day 14 was significantly reduced in the AM group and ES groups vs. the control group (33%, 40% and 100%, respectively, p = 0.03; mean esophageal diameter was 5.8±3.6 mm, 6.8±3.3 mm, and 2.6±1.7 mm for AM, ES, and control groups, respectively. Median (range esophageal fibrosis thickness was 0.87 mm (0.78-1.72, 1.19 mm (0.28-1.95, and 1.65 mm (0.7-1.79 for AM 1, ES, and control groups, respectively. All animals had developed esophageal strictures by day 35.The anti-fibrotic effect of AM on esophageal wound healing after ESD delayed the development of esophageal stricture in our model. However, this benefit was of limited duration in the conditions of our study.

  2. Evaluation of Amniotic Membrane Effectiveness in Skin Graft Donor Site Dressing in Burn Patients.

    Science.gov (United States)

    Salehi, Seyed Hamid; As'adi, Kamran; Mousavi, Seyed Jaber; Shoar, Saeed

    2015-12-01

    Although the recipient site in burn wounds is dressed with universally accepted materials, the ideal management of split-thickness skin donor sites remains controversial. The aim of our study is to compare two methods of wound dressing in donor sites of split-thickness skin graft in patients undergoing burn wound reconstructive surgery. Forty-two consecutive patients with second- and third-degree burns with a total body surface area between 20 and 40 % were enrolled in this randomized clinical trial conducted in Motahari Burn Hospital in Tehran, Iran. In each patient, two anatomic areas with similar features were randomly selected as intervention and control donor sites. The intervention site was dressed with amniotic membrane, whereas the control site was treated with Vaseline-impregnated gauze. Wounds were examined daily by expert surgeons to measure the clinical outcomes including duration of healing, severity of pain, and infection rate. The mean ± SD age of patients was 31.17 ± 13.72 years; furthermore, burn percentage had a mean ± SD of 31.19 ± 10.56. The mean ± SD of patients' cooperation score was 1.6 ± 0.79 in the intervention group compared with 2.93 ± 0.71 in the control group, revealing a statistically significant difference (P  0.05). Amniotic membrane as an alternative for dressing of skin graft donor sites provides significant benefits by increasing patients' comfort via diminishing the number of dressing changes and facilitating the process of wound healing.

  3. The use of Amniotic membrane in the treatment of Burns in Children: a clinical trial at the university Teaching Hospital, Lusaka

    International Nuclear Information System (INIS)

    Katebe, K.R.C

    1995-01-01

    This is a clinical trial which was carried out at the University Teaching Hospital, Lusaka from the 1st of July to the 31st December, 1994. It involved treatment of burns in forty children using gamma irradiated amniotic membrane produced at the hospital. The results showed that it is feasible to produce Gamma irradiated biological dressings from amniotic membrane at this hospital. The amniotic membrane was easy to apply on burns and the treatment was acceptable to the majority of parents with burnt children. The use of amniotic membrane was non inflammatory to the wounds in all forty patients (100%), reduced wound infection in thirty three patients (82.5%), increased the rate of wound healing in thirty nine patients (97.5%), and resulted in good quality wound healing in thirty one patients (77.5%). Therefore, the treatment offers a good alternative in the treatment of burns in children at the hospital

  4. IL-6 trans-signaling system in intra-amniotic inflammation, preterm birth, and preterm premature rupture of the membranes.

    Science.gov (United States)

    Lee, Sarah Y; Buhimschi, Irina A; Dulay, Antonette T; Ali, Unzila A; Zhao, Guomao; Abdel-Razeq, Sonya S; Bahtiyar, Mert O; Thung, Stephen F; Funai, Edmund F; Buhimschi, Catalin S

    2011-03-01

    Classic IL-6 signaling is conditioned by the transmembrane receptor (IL-6R) and homodimerization of gp130. During trans-signaling, IL-6 binds to soluble IL-6R (sIL-6R), enabling activation of cells expressing solely gp130. Soluble gp130 (sgp130) selectively inhibits IL-6 trans-signaling. To characterize amniotic fluid (AF) IL-6 trans-signaling molecules (IL-6, sIL-6R, sgp130) in normal gestations and pregnancies complicated by intra-amniotic inflammation (IAI), we studied 301 women during second trimester (n = 39), third trimester (n = 40), and preterm labor with intact (n = 131, 85 negative IAI and 46 positive IAI) or preterm premature rupture of membranes (PPROM; n = 91, 61 negative IAI and 30 positive IAI). ELISA, Western blotting, and real-time RT-PCR were used to investigate AF, placenta, and amniochorion for protein and mRNA expression of sIL-6R, sgp130, IL-6R, and gp130. Tissues were immunostained for IL-6R, gp130, CD15(+) (polymorphonuclear), and CD3(+) (T cell) inflammatory cells. The ability of sIL-6R and sgp130 to modulate basal and LPS-stimulated release of amniochorion matrix metalloprotease-9 was tested ex vivo. We showed that in physiologic gestations, AF sgp130 decreases toward term. AF IL-6 and sIL-6R were increased in IAI, whereas sgp130 was decreased in PPROM. Our results suggested that fetal membranes are the probable source of AF sIL-6R and sgp130. Immunohistochemistry and RT-PCR revealed increased IL-6R and decreased gp130 expression in amniochorion of women with IAI. Ex vivo, sIL-6R and LPS augmented amniochorion matrix metalloprotease-9 release, whereas sgp130 opposed this effect. We conclude that IL-6 trans-signaling molecules are physiologic constituents of the AF regulated by gestational age and inflammation. PPROM likely involves functional loss of sgp130.

  5. Reconstruction of limbal stem cell deficient corneal surface with induced human bone marrow mesenchymal stem cells on amniotic membrane.

    Science.gov (United States)

    Rohaina, Che Man; Then, Kong Yong; Ng, Angela Min Hwei; Wan Abdul Halim, Wan Haslina; Zahidin, Aida Zairani Mohd; Saim, Aminuddin; Idrus, Ruszymah B H

    2014-03-01

    The cornea can be damaged by a variety of clinical disorders or chemical, mechanical, and thermal injuries. The objectives of this study were to induce bone marrow mesenchymal stem cells (BMSCs) to corneal lineage, to form a tissue engineered corneal substitute (TEC) using BMSCs, and to treat corneal surface defects in a limbal stem cell deficiency model. BMSCs were induced to corneal lineage using limbal medium for 10 days. Induced BMSCs demonstrated upregulation of corneal stem cell markers; β1-integrin, C/EBPδ, ABCG2, and p63, increased protein expression of CK3 and p63 significantly compared with the uninduced ones. For TEC formation, passage 1 BMSCs were trypsinized and seeded on amniotic membrane in a transwell co-culture system and were grown in limbal medium. Limbal stem cell deficiency models were induced by alkaline injury, and the TEC was implanted for 8 weeks. Serial slit lamp evaluation revealed remarkable improvement in corneal regeneration in terms of corneal clarity and reduced vascularization. Histologic and optical coherence tomography analyses demonstrated comparable corneal thickness and achieved stratified epithelium with a compact stromal layer resembling that of normal cornea. CK3 and p63 were expressed in the newly regenerated cornea. In conclusion, BMSCs can be induced into corneal epithelial lineage, and these cells are viable for the formation of TEC, to be used for the reconstruction of the corneal surface in the limbal stem cell deficient model. Copyright © 2014 Mosby, Inc. All rights reserved.

  6. Mesenchymal Stem Cells from Human Amniotic Membrane and Umbilical Cord Can Diminish Immunological Response in an in vitro Allograft Model.

    Science.gov (United States)

    Dabrowski, Filip A; Burdzinska, Anna; Kulesza, Agnieszka; Chlebus, Marcin; Kaleta, Beata; Borysowski, Jan; Zolocinska, Aleksandra; Paczek, Leszek; Wielgos, Miroslaw

    2017-01-01

    Mesenchymal stem cells (MSCs) are gaining rising interest in gynecology and obstetrics. MSCs immunomodulatory properties are suitable enough to reduce perinatal morbidity caused by inflammation in premature neonates. The aim of this study was to evaluate and compare the ability to inhibit allo-activated lymphocytes proliferation by MSCs derived from different sources: amniotic membrane (AM), umbilical cord (UC) and adipose tissue (AT). MSCs were isolated from AM (n = 7) and UC (n = 6) and AT (n = 6) of healthy women. Cells were characterized by flow cytometry and differentiation assay. To evaluate the potential of fetal and adult MSCs to diminish immunological response, mixed lymphocytes reaction (MLR) was performed. Amnion and UC-derived cells displayed typical MSCs characteristics. Addition of MSCs to MLR significantly inhibited the proliferation of stimulated lymphocytes. The effect was observed regardless of the MSCs type used (p < 0.01 in all groups). Comparative analysis revealed no significant differences in this action between tested MSCs types. Additionally, no type of MSCs significantly stimulated allogeneic lymphocytes. The results prove the immunosuppressive capacities of fetal-derived MSCs in vitro. In the future, they may be potentially used to treat premature newborn as well as in immunomodulation in post-transplant therapy. © 2016 S. Karger AG, Basel.

  7. Radiosterilization of freeze-dried human amniotic Membrane and its use in the treatment of burn wound. Algerian experience

    International Nuclear Information System (INIS)

    Djefal, A.; Mahlous, M.; Nacer Khodja, A.; Larbi, M.; Larbi Daho Bachir, M.

    2001-01-01

    The present study evaluates the usefulness of human amniotic membrane as biological dressing and its efficacy in the treatment of burns comparatively to the conventional dressing. We reported the practical methods of preparation, preservation and radiation sterilisation of amnion, and the clinical results of its successful use in the treatment of 80 cases of superficial and intermediate depth dermal burns. The increased rate of healing, pain relief, good adhesion to the bed wound and absence of infection were observed

  8. Glutaraldehyde cross-linking of amniotic membranes affects their nanofibrous structures and limbal epithelial cell culture characteristics.

    Science.gov (United States)

    Lai, Jui-Yang; Ma, David Hui-Kang

    2013-01-01

    Given that the cells can sense nanometer dimensions, the chemical cross-linking-mediated alteration in fibrillar structure of collagenous tissue scaffolds is critical to determining their cell culture performances. This article explores, for the first time, the effect of nanofibrous structure of glutaraldehyde (GTA) cross-linked amniotic membrane (AM) on limbal epithelial cell (LEC) cultivation. Results of ninhydrin assays demonstrated that the amount of new cross-links formed between the collagen chains is significantly increased with increasing the cross-linking time from 1 to 24 hours. By transmission electron microscopy, the AM treated with GTA for a longer duration exhibited a greater extent of molecular aggregation, thereby leading to a considerable increase in nanofiber diameter and resistance against collagenase degradation. In vitro biocompatibility studies showed that the samples cross-linked with GTA for 24 hours are not well-tolerated by the human corneal epithelial cell cultures. When the treatment duration is less than 6 hours, the biological tissues cross-linked with GTA for a longer time may cause slight reductions in 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt, and anti-inflammatory activities. Nevertheless, significant collagen molecular aggregation also enhances the stemness gene expression, indicating a high ability of these AM matrices to preserve the progenitors of LECs in vitro. It is concluded that GTA cross-linking of collagenous tissue materials may affect their nanofibrous structures and corneal epithelial stem cell culture characteristics. The AM treated with GTA for 6 hours holds promise for use as a niche for the expansion and transplantation of limbal epithelial progenitor cells.

  9. Histological chorioamnionitis in preterm prelabor rupture of the membranes is associated with increased expression of galectin-3 by amniotic epithelium.

    Science.gov (United States)

    Stefanoska, Ivana; Tadić, Jasmina; Vilotić, Aleksandra; Jovanović Krivokuća, Milica; Abu Rabi, Tamara; Vićovac, Ljiljana

    2017-09-01

    Gal-3, which can regulate immune responses upon infection and inflammation, was not studied so far in intrauterine infection leading to preterm prelabor rupture of the membranes (PPROM), although gal-1 was reported to be implicated in the process. Gal-3 mRNA and protein expression in amnion and its changes during histological chorioamnionitis were studied here. Fetal membranes were obtained from women with PPROM with (n =15) and without histological chorioamnionitis (n =15) during second and third trimester. Immunohistochemical reactivity was evaluated semiquantitatively and analyzed using t-test. Galectin profile of amniotic epithelia was determined by polymerase chain reaction (PCR) and change assessed in gal-3 in PPROM with (n =5) or without histological chorioamnionitis (n =5) by real-time PCR. Human amniotic epithelium was found to express gal-1, gal-3, gal-7 and gal-8 mRNA. Gal-3 mRNA and protein is increased in fetal membranes and in the amniotic epithelium in patients with chorionamnionitis. Histological chorioamnionitis is associated with increased gal-3 expression and strong immunoreactivity of the amnion. Gal-3 may participate in the regulation of the inflammatory responses to chorioamniotic infection and/or direct interaction with pathogens.

  10. Allogeneic amniotic membrane-derived mesenchymal stromal cell transplantation in a porcine model of chronic myocardial ischemia

    Directory of Open Access Journals (Sweden)

    Kimura M

    2012-01-01

    Full Text Available Introduction. Amniotic membrane contains a multipotential stem cell population and is expected to possess the machinery to regulate immunological reactions. We investigated the safety and efficacy of allogeneic amniotic membrane-derived mesenchymal stromal cell (AMSC transplantation in a porcine model of chronic myocardial ischemia as a preclinical trial. Methods. Porcine AMSCs were isolated from amniotic membranes obtained by cesarean section just before delivery and were cultured to increase their numbers before transplantation. Chronic myocardial ischemia was induced by implantation of an ameroid constrictor around the left circumflex coronary artery. Four weeks after ischemia induction, nine swine were assigned to undergo either allogeneic AMSC transplantation or normal saline injection. Functional analysis was performed by echocardiography, and histological examinations were carried out by immunohistochemistry 4 weeks after AMSC transplantation. Results. Echocardiography demonstrated that left ventricular ejection fraction was significantly improved and left ventricular dilatation was well attenuated 4 weeks after AMSC transplantation. Histological assessment showed a significant reduction in percentage of fibrosis in the AMSC transplantation group. Injected allogeneic green fluorescent protein (GFP-expressing AMSCs were identified in the immunocompetent host heart without the use of any immunosuppressants 4 weeks after transplantation. Immunohistochemistry revealed that GFP colocalized with cardiac troponin T and cardiac troponin I. Conclusions. We have demonstrated that allogeneic AMSC transplantation produced histological and functional improvement in the impaired myocardium in a porcine model of chronic myocardial ischemia. The transplanted allogeneic AMSCs survived without the use of any immunosuppressants and gained cardiac phenotype through either their transdifferentiation or cell fusion.

  11. DNA-methylation profiling of fetal tissues reveals marked epigenetic differences between chorionic and amniotic samples.

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    Christel Eckmann-Scholz

    Full Text Available Epigenetic mechanisms including DNA methylation are supposed to play a key role in fetal development. Here we have investigated fetal DNA-methylation levels of 27,578 CpG loci in 47 chorionic villi (CVS and 16 amniotic cell (AC samples. Methylation levels differed significantly between karyotypically normal AC and CVS for 2,014 genes. AC showed more extreme DNA-methylation levels of these genes than CVS and the differentially methylated genes are significantly enriched for processes characteristic for the different cell types sampled. Furthermore, we identified 404 genes differentially methylated in CVS with trisomy 21. These genes were significantly enriched for high CG dinucleotid (CpG content and developmental processes associated with Down syndrome. Our study points to major tissue-specific differences of fetal DNA-methylation and gives rise to the hypothesis that part of the Down syndrome phenotype is epigenetically programmed in the first trimester of pregnancy.

  12. Term Amniotic membrane is a high throughput source for multipotent Mesenchymal Stem Cells with the ability to differentiate into endothelial cells in vitro

    DEFF Research Database (Denmark)

    Alviano, Francesco; Fossati, Valentina; Marchionni, Cosetta

    2007-01-01

    of CD34 and von Willebrand Factor positive cells. CONCLUSION: The current study suggests that AM-hMSCs may emerge as a remarkable tool for the cell therapy of multiple diseased tissues. AM-hMSCs may potentially assist both bone and cartilage repair, nevertheless, due to their angiogenic potential......BACKGROUND: Term Amniotic membrane (AM) is a very attractive source of Mesenchymal Stem Cells (MSCs) due to the fact that this fetal tissue is usually discarded without ethical conflicts, leading to high efficiency in MSC recovery with no intrusive procedures. Here we confirmed that term AM......, as previously reported in the literature, is an abundant source of hMSCs; in particular we further investigated the AM differentiation potential by assessing whether these cells may also be committed to the angiogenic fate. In agreement with the recommendation of the International Society for Cellular Therapy...

  13. Dried Human Amniotic Membrane Does Not Alleviate Inflammation and Fibrosis in Experimental Strabismus Surgery

    Directory of Open Access Journals (Sweden)

    Bo Young Chun

    2013-01-01

    Full Text Available Purpose. The purpose of this study was to evaluate the efficacy of dried human amniotic membrane (AM in reducing the postoperative inflammatory response and scarring after strabismus surgery. Methods. The inflammatory response at the extraocular muscle reattachment site was analyzed after superior rectus (SR resection in 12 rabbits. Dried human AM (Ambiodry2 was applied between the resected SR muscle plane and Tenon’s capsule of the left eyes of rabbits. As a control, the right eyes of rabbits underwent SR resection only. The surgeon randomly ordered which eye gets operated first during the experiment. Two weeks later, enucleation was performed. Six sagittal sections were made for each eye at the insertion of the SR muscle. The grade of postoperative inflammation and the presence of fibrosis were evaluated in histological examinations. Results. There was no statistically significant difference in the intensity of inflammation and fibrous proliferation between the eyes treated with dried human AM after SR resection and those treated with SR resection only. Conclusions. The use of dried human AM was not effective in controlling the postoperative inflammation and scarring in rabbit eyes after extraocular muscle surgery. However, this may be due to the devitalized dry preparation of human AM (Ambiodry2, which may have lost the expected anti-inflammatory and anti-scarring properties, and further studies on humans may be necessary.

  14. Clinical observation of cryotherapy and amniotic membrane transplantation in the management of Fusarium corneal ulcer

    Directory of Open Access Journals (Sweden)

    Qiong Xie

    2014-10-01

    Full Text Available AIM:To examine the efficacy and safety of cryotherapy in the management of Fusarium corneal ulcer. METHODS: Retrospective contract analysis of the clinical data of 41 patients(41 eyeswho infected with Fusarium corneal ulcer. All of them underwent focal lesion keratectomy combined with amniotic membrane transplantation between January 2010 and May 2013. The cryotherapy treatment group of 22 cases(22 eyes, non-cryotherapy group of 19 cases(19 eyes. All the cases were followed up for 3~12mo. We analyzed the healing of corneal ulcer, corneal neovascularization, postoperative visual acuity, complications and ulcer recurrences. RESULTS: There were no significant difference for the two groups at the cure rate and the recovery rate, the corneal neovascularization and postoperative visual acuity(P>0.05. The total effective rate and corneal healing time in cryotherapy group was significantly higher than that in non-cryotherapy group(PPCONCLUSION: Cryotherapy is an effective and safe method for the treatment of Fusarium corneal ulcer.

  15. Amniotic membrane extract ameliorates benzalkonium chloride-induced dry eye in a murine model.

    Science.gov (United States)

    Xiao, Xinye; Luo, Pingping; Zhao, Hui; Chen, Jingyao; He, Hui; Xu, Yuxue; Lin, Zhirong; Zhou, Yueping; Xu, Jianjiang; Liu, Zuguo

    2013-10-01

    Human amniotic membrane (AM) is avascular but contains various beneficial bioactive factors, its extract (AE) is also effective in treating many ocular surface disorders. In this study, we for the first time evaluated the therapeutic effects of AE on dry eye induced by benzalkonium chloride in a BALB/c mouse model. Topical application of AE (1.5 and 3 μg/eye/day) resulted in significantly longer tear break-up time on Day 3 and 6, lower fluorescein staining scores on Day 3, and lower inflammatory index on Day 6. AE reduced corneal epithelial K10 expression, inflammatory infiltration, and levels of TNF-α, IL-1β and IL-6 in BAC treated mice than that in the control mice. Moreover, decreased TUNEL positive cells in cornea and increased goblet cells in conjunctiva were also observed in AE treated corneas. Finally, AE induced more Ki-67 positive cells in corneal epithelium of dry eye mouse. Taken together, our data provide further support for BAC induced dry eye model as a valuable for dry eye study and suggest a great potential for AE as a therapeutic agent in the clinical treatment of dry eye. Copyright © 2013 Elsevier Ltd. All rights reserved.

  16. Augmented dried versus cryopreserved amniotic membrane as an ocular surface dressing.

    Directory of Open Access Journals (Sweden)

    Claire L Allen

    Full Text Available Dried amniotic membrane (AM can be a useful therapeutic adjunct in ophthalmic surgery and possesses logistical advantages over cryopreserved AM. Differences in preservation techniques can significantly influence the biochemical composition and physical properties of AM, potentially affecting clinical efficacy. This study was established to investigate the biochemical and structural effects of drying AM in the absence and presence of saccharide lyoprotectants and its biocompatibility compared to cryopreserved material.AM was cryopreserved or dried with and without pre-treatment with trehalose or raffinose and the antioxidant epigallocatechin (EGCG. Structural and visual comparisons were assessed using electron microscopy. Localisation, expression and release of AM biological factors were determined using immunoassays and immunofluorescence. The biocompatibility of the AM preparations co-cultured with corneal epithelial cell (CEC or keratocyte monolayers were assessed using cell proliferation, cytotoxicity, apoptosis and migration assays.Drying devitalised AM epithelium, but less than cryopreservation and cellular damage was reduced in dried AM pre-treated with trehalose or raffinose. Dried AM alone, and with trehalose or raffinose showed greater factor retention efficiencies and bioavailability compared to cryopreserved AM and demonstrated a more sustained biochemical factor time release in vitro. Cellular health assays showed that dried AM with trehalose or raffinose are compatible and superior substrates compared to cryopreserved AM for primary CEC expansion, with increased proliferation and reduced LDH and caspase-3 levels. This concept was supported by improved wound healing in an immortalised human CEC line (hiCEC co-cultured with dried and trehalose or raffinose membranes, compared to cryopreserved and fresh AM.Our modified preservation process and our resultant optimised dried AM has enhanced structural properties and biochemical stability

  17. Comparative evaluation of coronally advanced flap using amniotic membrane and platelet-rich fibrin membrane in gingival recession: An 18-month clinical study

    Directory of Open Access Journals (Sweden)

    Mohd Rehan

    2018-01-01

    Full Text Available Background: An amnion membrane is a placenta-derived tissue that consists of numerous growth factors, proteins, and stem cell reserves which help in accelerated wound healing and regeneration. Platelet-rich fibrin (PRF also releases growth factors after activation from the platelets and gets trapped within fibrin matrix which has been shown to stimulate the mitogenic response in the periosteum for bone repair and regeneration during normal wound healing. This preliminary, controlled, randomized clinical trial with an 18-month follow-up was aimed to evaluate the effectiveness of coronally advanced flap (CAF with either PRF membrane or bioresorbable amniotic membrane (AM in treatment of localized gingival recession defects. Materials and Methods: Sixteen healthy adult patients presenting with Miller Class I recession defects were treated surgically with CAF along with AM (Group I or PRF (Group II for coverage of the recession defects. For all patients, plaque index, gingival index, bleeding on probing, clinical attachment level, depth of recession, width of recession, width of attached gingiva, and gingival thickness were evaluated at 6 months and 18 months postoperatively. Statistical analysis was done using paired t-test, repeated measure analysis of variance test, Bonferroni test for intragroup comparison and unpaired t-test for intergroup comparison. Results: The results showed statistically nonsignificant (P < 0.01 difference in all clinical parameters at the 6- and 18-month follow-ups in both groups. Gingival recession in both PRF and amnion group when evaluated individually, significantly reduced from baseline to 6 months (P = 0.000 and from baseline to 18 months (P = 0.000. However, the mean value from 6 months to 18 months was statistically nonsignificant. Conclusion: The present study demonstrated that both CAF + PRF and CAF + AM are equally effective in providing clinically significant outcomes with respect to root coverage with AM

  18. Survey of Pregnancy Outcome in Preterm Premature Rupture of Membranes with Amniotic Fluid Index <5 and ≥5

    Directory of Open Access Journals (Sweden)

    Fatemeh Tavassoli

    2010-04-01

    Full Text Available Objectives: Preterm premature rupture of membranes (PPROM is among the most important causes of perinatal morbidity and mortality. The aim of this study is to survey the pregnancy outcomes in preterm premature rupture of membranes with an amniotic fluid index of 5.Methods: This prospective cohort study was performed on 137 pregnant women complicated by preterm premature rupture of membranes (PPROM with a gestational age of 28-34 weeks during October 2006 to October 2008. The patients were divided in two groups according to their amniotic fluid index; AFI<5 (77cases, AFI≥5 (60cases. The Chi-squared test for qualitative variables and T-student test for quantitative variables were used to analyze the results.Results: The results showed that there was no significant difference in terms of the number of pregnancies, gestational age at rupture of membranes and birthweight between the two groups. However, the results demonstrated that the patients with AFI<5 exhibited a significantly shorter latency period (p=0.049, a higher rate of cesarean due to fetal distress (p=0.008, a lower neonatal Apgar score in the first minute (p=0.0127 and a higher rate of neonatal death during the first week (p=0.045.Conclusion: Overall, PPROM with oligohydroamnios is associated with shorter latency, higher rate of C/S, higher rate of early neonatal death and lower neonatal Apgar.

  19. Clinical and impression cytology findings of amniotic membrane and oral mucosal membrane transplantation for the management of socket contracture.

    Science.gov (United States)

    Kurtul, Bengi Ece; Erdener, Ugur; Mocan, Mehmet Cem; Irkec, Murat; Orhan, Mehmet

    2014-01-01

    To investigate and compare the cytopathological and clinical effects of amniotic membrane transplantation (AMT) and oral mucosal membrane transplantation (OMMT) in socket contraction. Twelve patients who could not be fitted with ocular prosthesis due to socket contracture were included in this study. Seven patients underwent AMT and 5 patients underwent OMMT. Thirteen patients who had healthy sockets were included as control group. Depth of inferior fornix, degree of inflammation, extent of the socket contracture and socket volume were measured in the preoperative period and at sixth and twelfth weeks postoperatively. Impression cytology of conjunctival fornices and tear transforming growth factor beta-1 (TGFβ1) levels were determined. In the AMT group, socket volume and lower fornix depth values were significantly higher (P=0.030 and P=0.004 respectively) and inflammation levels and impression cytology stages (P=0.037 and P=0.022 respectively) were significantly lower in postoperative period compared to preoperative period. In the OMMT group, no statistical differences were found in terms of clinical parameters, inflammation levels and impression cytology stages of preoperative versus postoperative values. Preoperative tear TGFβ1 levels were higher in AMT and OMMT groups compared to the control group (25.5 ng/mL, 26.3 ng/mL and 21.7 ng/mL respectively). Decreased tear TGFβ1 levels were observed in both the AMT and OMMT groups postoperatively (median decrease value=2.1 ng/mL and 2.7 ng/mL respectively). AMT is associated with postoperative improvement in inferior fornix depth, socket volume, inflammation and impression cytology levels and may be a more proper alternative method than OMMT in the management of socket contracture.

  20. Corneal Nerve Regeneration after Self-Retained Cryopreserved Amniotic Membrane in Dry Eye Disease

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    Thomas John

    2017-01-01

    Full Text Available Purpose. To evaluate the efficacy of self-retained cryopreserved amniotic membrane (CAM in promoting corneal nerve regeneration and improving corneal sensitivity in dry eye disease (DED. Methods. In this prospective randomized clinical trial, subjects with DED were randomized to receive CAM (study group or conventional maximum treatment (control. Changes in signs and symptoms, corneal sensitivity, topography, and in vivo confocal microscopy (IVCM were evaluated at baseline, 1 month, and 3 months. Results. Twenty subjects (age 66.9 ± 8.9 were enrolled and 17 completed all follow-up visits. Signs and symptoms were significantly improved in the study group yet remained constant in the control. IVCM showed a significant increase in corneal nerve density in the study group (12,241 ± 5083 μm/mm2 at baseline, 16,364 ± 3734 μm/mm2 at 1 month, and 18,827 ± 5453 μm/mm2 at 3 months, p=0.015 but was unchanged in the control. This improvement was accompanied with a significant increase in corneal sensitivity (3.25 ± 0.6 cm at baseline, 5.2 ± 0.5 cm at 1 month, and 5.6 ± 0.4 cm at 3 months, p<0.001 and corneal topography only in the study group. Conclusions. Self-retained CAM is a promising therapy for corneal nerve regeneration and accelerated recovery of the ocular surface health in patients with DED. The study is registered at clinicaltrials.gov with trial identifier: NCT02764814.

  1. Cryopreserved Ultra-Thick Human Amniotic Membrane for Conjunctival Surface Reconstruction After Excision of Conjunctival Tumors.

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    Tanaka, Thais S; Demirci, Hakan

    2016-04-01

    Cryopreserved ultra-thick human amniotic membrane (AM) is used for glaucoma surgery. We evaluated the use of cryopreserved ultra-thick human AM for conjunctival surface reconstruction after excision of a conjunctival tumor. We retrospectively reviewed the medical records of 28 patients who underwent conjunctival surface reconstruction with cryopreserved ultra-thick human AM after excision of the tumor. The AM was secured to the surrounding conjunctiva and underlying sclera with interrupted 8-0 Vicryl sutures. Clinical data regarding demographics, diagnosis, size and location of conjunctival tumors, patient outcome, and complications were gathered. Of 28 patients, 6 (21.4%) had malignant melanoma, 4 (14.3%) had squamous cell carcinoma, 6 (21.4%) had conjunctival intraepithelial neoplasia, 1 (3.6%) had sebaceous carcinoma, 1 (3.6%) had mucoepidermoid carcinoma, 1 (3.6%) had conjunctival intraepithelial dysplasia, 5 (17.9%) had pterygium, 2 (7.1%) had compound nevus, 1 (3.6%) had a large epithelial inclusion cyst, and 1 (3.6%) patient had a granuloma. The mean area of graft size was 156 ± 120 mm2. Postoperatively, the graft was well tolerated with no failure, discomfort, or dehiscence. During the 17-month mean follow-up, symblepharon, which was clinically nonsignificant, developed in 3 (11%) patients and partial stem cell deficiency was noted in 5 (18%) patients. Cryopreserved ultra-thick human AM is a well-tolerated, effective graft material that is easy to handle. It is a viable alternative for conjunctival surface reconstruction after excision of a conjunctival tumor.

  2. Adjunctive Mitomycin C or Amniotic Membrane Transplantation for Ahmed Glaucoma Valve Implantation: A Randomized Clinical Trial.

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    Yazdani, Shahin; Mahboobipour, Hassan; Pakravan, Mohammad; Doozandeh, Azadeh; Ghahari, Elham

    2016-05-01

    To determine whether adjunctive mitomycin C (MMC) or amniotic membrane transplantation (AMT) improve the outcomes of Ahmed glaucoma valve (AGV) implantation. This double-blind, stratified, 3-armed randomized clinical trial includes 75 eyes of 75 patients aged 7 to 75 years with refractory glaucoma. Eligible subjects underwent stratified block randomization; eyes were first stratified to surgery in the superior or inferior quadrants based on feasibility; in each subgroup, eyes were randomly assigned to the study arms using random blocks: conventional AGV implantation (group A, 25 eyes), AGV with MMC (group B, 25 eyes), and AGV with AMT (group C, 25 eyes). The 3 study groups were comparable regarding baseline characteristics and mean follow-up (P=0.288). A total of 68 patients including 23 eyes in group A, 25 eyes in group B, and 20 eyes group C completed the follow-up period and were analyzed. Intraocular pressure was lower in the MMC group only 3 weeks postoperatively (P=0.04) but comparable at other time intervals. Overall success rate was comparable in the 3 groups at 12 months (P=0.217). The number of eyes requiring medications (P=0.30), time to initiation of medications (P=0.13), and number of medications (P=0.22) were comparable. Hypertensive phase was slightly but insignificantly more common with standard surgery (82%) as compared with MMC-augmented (60%) and AMT-augmented (70%) procedures (P=0.23). Complications were comparable over 1 year (P=0.28). Although adjunctive MMC and AMT were safe during AGV implantation, they did not influence success rates or intraocular pressure outcomes. Complications, including hypertensive phase, were also comparable.

  3. Amnioinfusion in labor induction of term pregnancies with premature rupture of the membranes and low amniotic fluid.

    Science.gov (United States)

    Miño, M; Puertas, A; Herruzo, A J; Miranda, J A

    1998-05-01

    To analyze the utility of prophylactic amnioinfusion in term pregnancies with PROM and a low amniotic fluid index during labor induction. Forty-two women with amnioinfusion and 42 in a control group with amniotic fluid index (AFI) below 10 cm when admitted to labor induction were studied. All patients had electronic fetal heart rate and intrauterine pressure continuous monitoring. Amnioinfusion of normal saline (37 degrees C) was realized in the study group, using a continuous perfusion pump at 600 ml/h for 1 h, after which the AFI was again recorded; if this was amnioinfusion. Both groups were similar in age, primiparity, gestational age, initial AFI, interval from rupture of membranes until delivery and length of labor. The amnioinfusion of 600 ml in 1 h significantly increased the AFI (an increase of 7.2 +/- 3.9 vs. a decrease of 1.1 +/- 1.6, P amnioinfusion group, there was a significantly lower rate of cesarean deliveries (0 vs. 6, P amnioinfusion improves neonatal metabolic state when used in labor induction of term pregnancies with PROM and a low amniotic fluid index.

  4. Biocompatibility of quantum dots (CdSe/ZnS ) in human amniotic membrane-derived mesenchymal stem cells in vitro.

    Science.gov (United States)

    Wang, Gongping; Zeng, Guangwei; Wang, Caie; Wang, Huasheng; Yang, Bo; Guan, Fangxia; Li, Dongpeng; Feng, Xiaoshan

    2015-06-01

    Amniotic membrane-derived mesenchymal stem cells (hAM-dMSCs) are a potential source of mesenchymal stem cells which could be used to repair skin damage. The use of mesenchymal stem cells to repair skin damage requires safe, effective and biocompatible agents to evaluate the effectiveness of the result. Quantum dots (QDs) composed of CdSe/ZnS are semiconductor nanocrystals with broad excitation and narrow emission spectra, which have been considered as a new chemical and fluorescent substance for non-invasively labeling different cells in vitro and in vivo. This study investigated the cytotoxic effects of QDs on hAM-dMSCs at different times following labeling. Using 0.75, 1.5 and 3.0 μL between quantum dots, labeled human amniotic mesenchymal stem cells were collected on days 1, 2 and 4 and observed morphological changes, performed an MTT cell growth assay and flow cytometry for mesenchymal stem cells molecular markers. Quantum dot concentration 0.75 μg/mL labeled under a fluorescence microscope, cell morphology was observed, The MTT assay showed cells in the proliferative phase. Flow cytometry expression CD29, CD31, CD34, CD44, CD90, CD105 and CD106. Within a certain range of concentrations between quantum dots labeled human amniotic mesenchymal stem cells has good biocompatibility.

  5. Value of amniotic fluid IL-8 and Annexin A2 in prediction of preterm delivery in preterm labor and preterm premature rupture of membranes.

    Science.gov (United States)

    Jia, Xiaohui

    2014-01-01

    To investigate the clinical significance and value in the prediction of preterm delivery of combined amniotic fluid IL-8 and Annexin A2 levels in preterm premature rupture of membranes (PPROM) and preterm labor (PTL). Sixty pregnant women at < 32 gestational weeks who developed PTL were divided into a PPROM group and a non-PPROM group. Ten normal pregnant women served as a control group. IL-8 and Annexin A2 levels were measured in amniotic fluid samples from each patient. Amniotic fluid IL-8 and Annexin-A2 levels in PTL (PPROM and non-PPROM groups) were significantly higher than those of the controls (p < 0.05). The PPROM group displayed higher amniotic fluid Annexin-A2 levels than did the non-PPROM group, with a statistically significant difference (p < 0.05). The PPROM group showed higher amniotic fluid IL-8 levels than did the non-PPROM group; however, this was statistically insignificant (p = 0.56). Combined detection of amniotic fluid IL-8 and Annexin-A2 in the prediction of preterm delivery within 2 weeks of measurement showed sensitivity of 81.25%, specificity of 88.89% and PPV of 92.86%. Amniotic fluid IL-8 and Annexin-A2 levels are associated with the occurrence of PPROM and PTL. Combined detection of IL-8 and Annexin-A2 levels in identifying preterm delivery within 2 weeks in PTL and PPROM is of possible clinical and predictive value.

  6. Group B streptococcal infection of the choriodecidua induces dysfunction of the cytokeratin network in amniotic epithelium: a pathway to membrane weakening.

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    Jeroen P Vanderhoeven

    2014-03-01

    Full Text Available Early events leading to intrauterine infection remain poorly defined, but may hold the key to preventing preterm delivery. To determine molecular pathways within fetal membranes (chorioamnion associated with early choriodecidual infection that may progress to preterm premature rupture of membranes (PPROM, we examined the effects of a Group B Streptococcus (GBS choriodecidual infection on chorioamnion in a nonhuman primate model. Ten chronically catheterized pregnant monkeys (Macaca nemestrina at 118-125 days gestation (term = 172 days received choriodecidual inoculation of either GBS (n = 5 or saline (n = 5. Cesarean section was performed in the first week after GBS or saline inoculation. RNA extracted from chorioamnion (inoculation site was profiled by microarray. Single gene, Gene Set, and Ingenuity Pathway Analysis results were validated using qRT-PCR (chorioamnion, Luminex (amniotic fluid, AF, immunohistochemistry, and transmission electron microscopy (TEM. Despite uterine quiescence in most cases, significant elevations of AF cytokines (TNF-α, IL-8, IL-1β, IL-6 were detected in GBS versus controls (p2-fold change, p<0.05. Remarkably, GBS exposure was associated with significantly downregulated expression of multiple cytokeratin (CK and other cytoskeletal genes critical for maintenance of tissue tensile strength. Immunofluorescence revealed highly significant changes in the CK network within amniocytes with dense CK aggregates and retraction from the cell periphery (all p = 0.006. In human pregnancies affected by PPROM, there was further evidence of CK network retraction with significantly shorter amniocyte foot processes (p = 0.002. These results suggest early choriodecidual infection results in decreased cellular membrane integrity and tensile strength via dysfunction of CK networks. Downregulation of CK expression and perturbations in the amniotic epithelial cell intermediate filament network occur after GBS

  7. Prophylactic transabdominal amnioinfusion in oligohydramnios for preterm premature rupture of membranes: increase of amniotic fluid index during latency period.

    Science.gov (United States)

    Garzetti, G G; Ciavattini, A; De Cristofaro, F; La Marca, N; Arduini, D

    1997-01-01

    This study was designed to: (i) evaluate the effect of amnioinfusion on the latency period in patients with oligohydramnios for preterm premature rupture of membranes, and (ii) to investigate the relationship between changes in the amniotic fluid index and fetal heart rate short-term variability by computerized Hewlett-Packard cardiotocography, longitudinally estimated before and after prophylactic amnioinfusion. All singleton pregnancies with prolonged premature rupture of membranes after 25 weeks of gestation and seen at the Institute of Obstetrics and Gynecology, University of Ancona (Italy), between January 1994 and June 1995 were included in the study. Transabdominal amnioinfusion with 150-350 ml warmed normal saline (25-50 ml/min) was performed at weekly intervals. Amniotic fluid volume was assessed ultrasonographically by means of the four-quadrant technique on a weekly basis before and after each amnioinfusion, as well as the short-term variability by a Hewlett-Packard computerized cardiotocographic system. 18 women were enrolled and underwent prophylactic transabdominal amnioinfusion at weekly intervals until delivery. Eighteen controls, who did not undergo prophylactic amnioinfusion, were recruited from our 1992-1993 series and included in the study. The median interval between premature rupture of membranes and delivery was 3.0 weeks (range 1-8 weeks), with an average delivery age of 33.0 weeks (range 27-36 weeks). The latency period was significantly longer in patients who underwent prophylactic amnioinfusion (mean +/- SD, 4.1 +/- 1.7 weeks) than in controls(1.7 +/- 1.0 weeks; p amnioinfusion. A direct relationship was observed between the amniotic fluid index and short-term variability (linear regression analysis r = 0.54, p = 0.04). The mean values of fetal movements recorded by computerized tomography during the 20 min of observation significantly increased after amnioinfusion in comparison with those before it (2.6 +/- 0.9 and 0.9 +/- 0

  8. Comparative Study of Pregnancy Outcome in Premature Rupture of Membranes With Amniotic Fluid Indices of less and more than 5

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    F Tavassoli

    2010-03-01

    Full Text Available Introduction: Preterm premature rupture of the membranes (PPROM is one of the most important causes of perinatal morbidity and mortality. The aim of this study was to evaluate pregnancy outcome in preterm premature rupture of the membranes with amniotic fluid index less than 5 and more than 5. Methods: This prospective study was performed on 137 pregnant women with gestational age of 28-34 weeks with PPROM from Oct 2006 to Oct 2008. The patients were divided in two groups according to amniotic fluid index: AFI<5 (77 cases, AFI≥5 (60 cases. Chi-square test was used for qualitative variables, while T-student test was used for quantitative test. Results: Both groups were similar with respect to number of pregnancies, gestational age at rupture of the membranes and birth weight. In the group with AFI<5, applied latency was significantly shorter (PV=0.049, rate of cesarean section was higher due to fetal distress (PV=0.008, neonatal Apgar score in first minute was lower during the first week (PV=0.0127 and the rate of neonatal death was higher (PV=0.045 during the first week. Conclusion: An AFI<5 cm after PPROM is associated with earlier delivery, higher rate of cesarean due to fetal distress, higher rate of neonatal death, and lower neonatal Apgar score in first minute during the first week

  9. Human amniotic membrane, best healing accelerator, and the choice of bone induction for vestibuloplasty technique (an animal study

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    Ahad Khoshzaban

    2010-12-01

    Full Text Available Mohammad H Samandari1, Shahriar Adibi2, Ahad Khoshzaban3, Sara Aghazadeh5, Parviz Dihimi4, Siamak S Torbaghan6, Saeed H Keshel5, Zohreh Shahabi71Department of Oral and Maxillofacial Surgery, Dentistry Faculty, 2Dental Research of Torabinejad Research Centre, 3Iranian Tissue Bank Research and Preparation Centre, Imam Khomeini Hospital Complex, 4Department of Oral and Maxillofacial Pathology, Dentistry Faculty, Isfahan University of Medical Sciences, Isfahan, Iran; 5Stem Cells Preparation Unit, Eye Research Center, Farabi Hospital, 6Department of Pathology, Imam Khomeini Medical Centre, 7BMT Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, IranObjective: To investigate the effects of amniotic membrane (AM in bone induction and wound healing after vestibuloplasty surgery on animal samples while receptacle proteins such as growth factors were considered as accelerators for wound healing and bone induction after these operations.Material and methods: Ten adult dogs (5 females, 5 males; race, Iranian mixed; weight, 44 pounds were included, which underwent surgery for transplantation on mandible and maxillary. AM was used for promoting bone induction and healing.Results: The tissue samples were obtained after 2, 8, and 12 weeks for histology survey. No significant differences were observed between male and female or left and right jaws. AM decreased fibrinoleukocytic exudates and inflammation in the experimental group, had significant effects on bone formation, considerably improves wound healing, and gives rise to bone induction (P < 0.0001.Conclusions: Our study findings indicate that the AM is a suitable cover for different injuries and acellular AM has the potential for rapid improvement and bone induction. The AM contains collagen, laminin, and fibronectin, which provide an appropriate substrate for bone induction. This substrate promoted bone induction and might contribute to induction of the progenitor cells and/or stem

  10. Grafting with Cryopreserved Amniotic Membrane versus Conservative Wound Care in Treatment of Pressure Ulcers: A Randomized Clinical Trial.

    Science.gov (United States)

    Dehghani, Mehdi; Azarpira, Negar; Mohammad Karimi, Vahid; Mossayebi, Hamid; Esfandiari, Elaheh

    2017-10-01

    To compare the healing process of pressure ulcers treated with cryopreserved human amniotic membrane allograft and routine pressure ulcer care in our hospital. From January 2012 to December 2013, in a prospective randomized clinical trial (IRCT201612041335N2), 24 patients with second and third stage of pressure ulcers were enrolled in this study. All patients needed split-thickness skin grafts for pressure ulcer-wound coverage. Selected patients had symmetric ulcers on both upper and lower extremities. The patients were randomly divided into two groups: amnion and control. In the amnion group, the ulcer was covered with cryopreserved amniotic membrane and in the control group it was treated with local Dilantin powder application. The duration and success rate of complete healing was compared between the two groups. The study group was composed of 24 pressure ulcers in 24 patients (19 males and 5 females) with a mean age of 44±12.70 years. The demographic characteristics, ulcer area, and underlying diseases were similar in both groups. The early sign of response, such as decrease in wound discharge, was detected 12-14 days after biological dressing. Complete pressure ulcer healing occurred only in the amnion group ( p pressure ulcers.

  11. Transplantation with cultured stem cells derived from the human amniotic membrane for corneal alkali burns: an experimental study.

    Science.gov (United States)

    Zeng, Wei; Li, Yanwei; Zeng, Guangwei; Yang, Bo; Zhu, Yu

    2014-01-01

    Amniotic membranes (AM) have been used in a wide range of clinical applications. We successfully extracted mesenchymal stem cells (MSCs) from human AM, but little is known about the use and efficacy of human amniotic membrane-derived mesenchymal stem cells (hAM-dMSCs) for the treatment of alkali burns. We utilized hAM-dMSCs transplantation, AM grafting, and their combined use in the treatment of alkali burns. An experimental model in rabbits was devised to analyze the use of these techniques with immunocytochemistry and ELISA. The survival and migration of hAM-dMSCs labeled by SPION in the host were assessed with Prussian blue staining. Compared with the control group, the treated groups demonstrated faster reconstruction of the corneal epithelium, and lower levels of corneal opacification and neovascularization within corneal alkali burns. Furthermore, dark blue-stained particles were detected in the limbus corneae at day 28. These results demonstrated the ability of hAM-dMSCs to enhance epithelial healing and reduce corneal opacification and neovascularization in corneal alkali wounds.

  12. Maternal serum C-reactive protein concentration and intra-amniotic inflammation in women with preterm prelabor rupture of membranes.

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    Ivana Musilova

    Full Text Available To evaluate maternal serum C-reactive protein (CRP concentrations in pregnancies complicated by preterm prelabor rupture of membranes (PPROM in relation to the presence of microbial invasion of the amniotic cavity (MIAC and/or intra-amniotic inflammation (IAI.Two hundred and eighty-seven women with singleton pregnancies complicated by PPROM between 2014 and 2016 were included in this study. Maternal blood and amniotic fluid samples were collected at the time of admission. Maternal serum CRP concentration was measured using a high-sensitivity immunoturbidimetric assay. Interleukin-6 (IL-6 concentration was measured using a point-of-care test. MIAC was diagnosed based on a positive polymerase chain reaction result for Ureaplasma species, Mycoplasma hominis, and/or Chlamydia trachomatis and for the 16S rRNA gene. IAI was characterized by an amniotic fluid IL-6 concentration of ≥ 745 pg/mL.Women with MIAC and IAI had higher maternal serum CRP concentrations than did women without (with MIAC: median 6.9 mg/L vs. without MIAC: median 4.9 mg/L; p = 0.02; with IAI: median 8.6 mg/L vs. without IAI: median 4.7 mg/L; p < 0.0001. When women were split into four subgroups based on the presence of MIAC and/or IAI, women with the presence of both MIAC and IAI had higher maternal serum CRP than did women with IAI alone, with MIAC alone, and women without MIAC and IAI (both MIAC and IAI: median: 13.1 mg/L; IAI alone: 6.0 mg/L; MIAC alone: 3.9 mg/L; and without MIAC and IAI: median 4.8 mg/L; p < 0.0001. The maternal serum CRP cutoff value of 17.5 mg/L was the best level to identify the presence of both MIAC and IAI, with sensitivity of 47%, specificity of 96%, positive predictive value of 42%, negative predictive value of 96%, and the positive likelihood ratio of 10.9.The presence of both MIAC and IAI was associated with the highest maternal serum CRP concentrations. Maternal serum CRP concentration in women with PPROM at the time of admission can rule out the

  13. Evaluation of Human Amniotic Membrane as a Wound Dressing for Split-Thickness Skin-Graft Donor Sites

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    Denys J. Loeffelbein

    2014-01-01

    Full Text Available Human amniotic membrane (HAM has been used as a biomaterial in various surgical procedures and exceeds some qualities of common materials. We evaluated HAM as wound dressing for split-thickness skin-graft (STSG donor sites in a swine model (Part A and a clinical trial (Part B. Part A: STSG donor sites in 4 piglets were treated with HAM or a clinically used conventional polyurethane (PU foil (n=8 each. Biopsies were taken on days 5, 7, 10, 20, 40, and 60 and investigated immunohistochemically for alpha-smooth muscle actin (αSMA: wound contraction marker, von Willebrand factor (vWF: angiogenesis, Ki-67 (cell proliferation, and laminin (basement membrane integrity. Part B: STSG donor sites in 45 adult patients (16 female/29 male were treated with HAM covered by PU foam, solely by PU foam, or PU foil/paraffin gauze (n=15 each. Part A revealed no difference in the rate of wound closure between groups. HAM showed improved esthetic results and inhibitory effects on cicatrization. Angioneogenesis was reduced, and basement membrane formation was accelerated in HAM group. Part B: no difference in re-epithelialization/infection rate was found. HAM caused less ichor exudation and less pruritus. HAM has no relevant advantage over conventional dressings but might be a cost-effective alternative.

  14. Clinical significance of histologic chorioamnionitis with a negative amniotic fluid culture in patients with preterm labor and premature membrane rupture.

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    Jeong Woo Park

    Full Text Available To evaluate the effect of histological chorioamnionitis (HCA with a negative amniotic fluid (AF culture on adverse pregnancy and neonatal outcomes and inflammatory status in the AF compartment in women with preterm labor or preterm premature rupture of membranes (PPROM.This is a retrospective cohort study of 153 women diagnosed as having a preterm labor or PPROM (20-34 weeks who delivered singleton gestations within 48 hours of amniocentesis. AF obtained through amniocentesis was cultured, and interleukin (IL-6, IL-8, and metalloproteinase-9 (MMP-9 levels were determined. The placentas were examined histologically.The prevalence of HCA with negative AF culture was 23.5% (36/153. The women with HCA but with a negative AF culture (group 2 and those with a positive AF culture (group 3 had a significantly lower mean gestational age at amniocentesis and delivery than those with a negative AF culture and without HCA (group 1. Women in group 3 had the highest levels of AF IL-6, IL-8, and MMP-9, followed by those in group 2, and those in group 1. Composite neonatal morbidity was significantly higher in groups 2 and 3 than in group 1, but this was no longer significant after adjusting for confounders caused mainly by the impact of gestational age.In the women who delivered preterm neonates, HCA with a negative AF culture was associated with increased risks of preterm birth, intense intra-amniotic inflammatory response, and prematurity-associated composite neonatal morbidity, and its risks are similar to the risk posed by positive AF culture.

  15. Resorbable and running suture for stable fixation of amniotic membrane multilayers: A useful modification in deep or perforating sterile corneal ulcers

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    Constantin E. Uhlig

    2018-06-01

    Full Text Available Purpose: To present a modified technique for secure tightening and fixing of multilayer amniotic membranes in deep or perforating corneal ulcers. Observations: The modified procedure for application and fixation of multilayer amniotic membranes is retrospectively described step by step, and the results of three patients treated with this technique were retrospectively analysed and presented.The modification consists basically in fixing the inlays with one mini-overlay that is sutured intracorneally with resorbable and running Vicryl 10.0, before a corneoscleral overlay is fixed on top conjunctivally with a running nylon 10.0 suture. The resorbable Vicryl suture is left in place permanently. Conclusions and Importance: The method described avoids any risk of destroying or displacing the inlays by removing sutures later. In each of the three patients demonstrated as case reports the cornea remained stable throughout the 3- to 5-month follow-up period. This modified technique represents a very useful auxiliary means of treating deep or perforating non-infectious corneal ulcers. Keywords: Amniotic membrane transplantation, Corneal ulcer, Corneal perforation, Multilayer, Keratoplasty

  16. Modulation of amniotic fluid activin-a and inhibin-a in women with preterm premature rupture of the membranes and infection-induced preterm birth.

    Science.gov (United States)

    Rosenberg, Victor A; Buhimschi, Irina A; Dulay, Antonette T; Abdel-Razeq, Sonya S; Oliver, Emily A; Duzyj, Christina M; Lipkind, Heather; Pettker, Christian M; Buhimschi, Catalin S

    2012-02-01

    Activins and inhibins are important modulators of inflammatory processes. We explored activation of amniotic fluid (AF) activin-A and inhibin-A system in women with intra-amniotic infection and preterm premature rupture of the membranes (PPROM). We analyzed 78 AF samples: '2nd trimester-control' (n=12), '3rd trimester-control' (n=14), preterm labor with intact membranes [positive-AF-cultures (n=13), negative-AF-cultures (n=13)], and PPROM [positive-AF-cultures (n=13), negative-AF-cultures (n=13)]. Activin-A levels were evaluated ex-vivo following incubation of amniochorion and placental villous explants with Gram-negative lipopolysaccharide (LPS) or Gram-positive (Pam3Cys) bacterial mimics. Ability of recombinant activin-A and inhibin-A to modulate inflammatory reactions in fetal membranes was explored through explants' IL-8 release. Activin-A and inhibin-A were present in human AF and were gestational age-regulated. Activin-A was significantly upregulated by infection. Lower inhibin-A levels were seen in PPROM. LPS elicited release of activin-A from amniochorion, but not from villous explants. Recombinant activin-A stimulated IL-8 release from amniochorion, an effect that was not reversed by inhibin-A. Human AF activin-A and inhibin-A are involved in biological processes linked to intra-amniotic infection/inflammation-induced preterm birth. © 2011 John Wiley & Sons A/S.

  17. Electrospun polyurethane membranes for Tissue Engineering applications

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    Gabriel, Laís P., E-mail: lagabriel@gmail.com [National Institute of Biofabrication, Campinas (Brazil); Department of Chemical Engineering, University of Campinas, Campinas (Brazil); Rodrigues, Ana Amélia [National Institute of Biofabrication, Campinas (Brazil); Department of Medical Sciences, University of Campinas, Campinas (Brazil); Macedo, Milton; Jardini, André L.; Maciel Filho, Rubens [National Institute of Biofabrication, Campinas (Brazil); Department of Chemical Engineering, University of Campinas, Campinas (Brazil)

    2017-03-01

    Tissue Engineering proposes, among other things, tissue regeneration using scaffolds integrated with biological molecules, growth factors or cells for such regeneration. In this research, polyurethane membranes were prepared using the electrospinning technique in order to obtain membranes to be applied in Tissue Engineering, such as epithelial, drug delivery or cardiac applications. The influence of fibers on the structure and morphology of the membranes was studied using scanning electron microscopy (SEM), the structure was evaluated by Fourier transform infrared spectroscopy (FT-IR), and the thermal stability was analyzed by thermogravimetry analysis (TGA). In vitro cells attachment and proliferation was investigated by SEM, and in vitro cell viability was studied by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays and Live/Dead® assays. It was found that the membranes present an homogeneous morphology, high porosity, high surface area/volume ratio, it was also observed a random fiber network. The thermal analysis showed that the membrane degradation started at 254 °C. In vitro evaluation of fibroblasts cells showed that fibroblasts spread over the membrane surface after 24, 48 and 72 h of culture. This study supports the investigation of electrospun polyurethane membranes as biocompatible scaffolds for Tissue Engineering applications and provides some guidelines for improved biomaterials with desired properties.

  18. No midterm advantages in the middle term using small intestinal submucosa and human amniotic membrane in Achilles tendon transverse tenotomy.

    Science.gov (United States)

    Liu, Yushu; Peng, Yinbo; Fang, Yong; Yao, Min; Redmond, Robert W; Ni, Tao

    2016-11-24

    The study was aimed to compare the effects of small intestinal submucosa (SIS) and human amniotic membrane (HAM) on Achilles tendon healing. A total of 48 New Zealand white rabbits were divided into two groups. A full-thickness transverse tenotomy was made at the right leg of the rabbits. Then, the laceration site was wrapped with HAM (P/A group) or SIS (P/S group). The ultimate stress (US) and Young's modulus (E) of the tendons were detected for biomechanical analysis. Histological evaluation was performed using hematoxylin and eosin, immunohistochemical, and immunofluorescent stain. Expression of collagen I was detected by western blot analysis, and levels of inflammatory cytokines IL-1β, IL-6, and TNF-α were measured. Finally, adhesion formation was evaluated. There were no significant differences in filamentous adhesion, cross-sectional areas of the laceration sites, levels of inflammatory response, and collagen type I expression between the P/A and P/S groups (p > 0.05). Compared with the P/A group, the US and E values were significantly higher in the P/S group at day 7 (p Achilles tendon injury in the early stage of healing.

  19. Clinical observation on bandage type corneal contact lens applied after pterygium excision combined with fresh amniotic membrane transplantation

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    Bin Zhang

    2018-04-01

    Full Text Available AIM: To evaluate the clinical effect of fresh amniotic membrane transplantation in combination with bandage type corneal contact lens in the treatment of pterygium. METHODS: This study was a prospective analysis of 600 pterygium excision patients(600 cases, 600 eyesin the He Eye Hospital from January 2016 to June 2016. The patients were randomly and evenly divided into two groups: observation group and control group, 300 cases in each group(300 eyes. Observation group wore the bandage type corneal contact lens after surgery and took it down at fifth day. The control group did not. The degree of pain after surgery were evaluated at 2h, 1, 3 and 5d after surgery. The clinical outcomes in these two groups were compared in a 12mo postoperative follow-up observation. RESULTS: There were no significant differences between the observation and control group in the percentage of age and gender and size of the pterygium(P>0.05. After surgery, the degree of pain of treatment group significantly decreased compared to control group, the difference was statistically significant(PP>0.05. CONCLUSION: Bandage contact lenses could significantly release pain response after pterygium excision. But it can not reduce the recurrence rate.

  20. Skin Mast Cell Promotion in Random Skin Flaps in Rats using Bone Marrow Mesenchymal Stem Cells and Amniotic Membrane

    Science.gov (United States)

    Chehelcheraghi, Farzaneh; Abbaszadeh, Abolfazl; Tavafi, Magid

    2018-03-06

    Skin flap procedures are employed in plastic surgery, but failure can lead to necrosis of the flap. Studies have used bone marrow mesenchymal stem cells (BM-MSCs) to improve flap viability. BM-MSCs and acellular amniotic membrane (AAM) have been introduced as alternatives. The objective of this study was to evaluate the effect of BM-MSCs and AAM on mast cells of random skin flaps (RSF) in rats. RSFs (80 × 30 mm) were created on 40 rats that were randomly assigned to one of four groups, including (I) AAM, (II) BM-MSCs, (III) BM-MSCs/AAM, and (IV) saline (control). Transplantation was carried out during the procedure (zero day). Flap necrosis was observed on day 7, and skin samples were collected from the transition line of the flap to evaluate the total number and types of mast cells. The development and the total number of mast cells were related to the development of capillaries. The results of one-way ANOVA indicated that there was no statistically significant difference between the mean numbers of mast cell types for different study groups. However, the difference between the total number of mast cells in the study groups was statistically significant (p = 0.001). The present study suggests that the use of AAM/BM-MSCs can improve the total number of mast cells and accelerate the growth of capillaries at the transient site in RSFs in rats.

  1. Amniotic fluid embolism

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    Kiranpreet Kaur

    2016-01-01

    Full Text Available Amniotic fluid embolism (AFE is one of the catastrophic complications of pregnancy in which amniotic fluid, fetal cells, hair, or other debris enters into the maternal pulmonary circulation, causing cardiovascular collapse. Etiology largely remains unknown, but may occur in healthy women during labour, during cesarean section, after abnormal vaginal delivery, or during the second trimester of pregnancy. It may also occur up to 48 hours post-delivery. It can also occur during abortion, after abdominal trauma, and during amnio-infusion. The pathophysiology of AFE is not completely understood. Possible historical cause is that any breach of the barrier between maternal blood and amniotic fluid forces the entry of amniotic fluid into the systemic circulation and results in a physical obstruction of the pulmonary circulation. The presenting signs and symptoms of AFE involve many organ systems. Clinical signs and symptoms are acute dyspnea, cough, hypotension, cyanosis, fetal bradycardia, encephalopathy, acute pulmonary hypertension, coagulopathy etc. Besides basic investigations lung scan, serum tryptase levels, serum levels of C3 and C4 complements, zinc coproporphyrin, serum sialyl Tn etc are helpful in establishing the diagnosis. Treatment is mainly supportive, but exchange transfusion, extracorporeal membrane oxygenation, and uterine artery embolization have been tried from time to time. The maternal prognosis after amniotic fluid embolism is very poor though infant survival rate is around 70%.

  2. Effectiveness of the radio sterilized amniotic membrane transplantation vs conjunctival auto graft implant in the pterygium surgery with intraoperative mitomycin C;Eficacia del trasplante de membrana amniotica radioesterilizada vs injerto autologo conjuntival en la cirugia de pterigion con mitocomina C intraoperatoria

    Energy Technology Data Exchange (ETDEWEB)

    Tellez Y, L.; Martinez P, M. E. [ININ, Banco de Tejidos Radioesterilizados, Carretera Mexico-Toluca s/n, Ocoyoacac 52750, Estado de Mexico (Mexico); Vazquez M, L. [Hospital General de Mexico, Departamento de Cornea, Dr. Balmis No. 148, Col. Doctores, 06726 Mexico D. F. (Mexico)

    2009-07-01

    At the present time the traditional surgical handling of the pterygium with conjunctival implant presents high frequency. In this work the obtained results of a controlled clinic practice blind double are presented of the period of December from 2008 to December 2009, realized in the Medical Unit of Ambulatory Attention 231 of the Mexican Institute of the Public Health located in Metepec, Mexico State, in order to demonstrate the effectiveness and security of the alternative use of the radio sterilized amniotic membrane and intraoperative mitomycin C. For the study was included patients with diagnostic of primary nasal pterygium in one or both eyes, adults of both genus with an age range of 20 to 60 years, and was used radio sterilized amniotic membrane processed in the Bank of Radio Sterilized Tissues of the National Institute of Nuclear Research. (Author)

  3. Different therapeutic effects of cells derived from human amniotic membrane on premature ovarian aging depend on distinct cellular biological characteristics.

    Science.gov (United States)

    Ding, Chenyue; Li, Hong; Wang, Yun; Wang, Fuxin; Wu, Huihua; Chen, Rulei; Lv, Jinghuan; Wang, Wei; Huang, Boxian

    2017-07-27

    Many reports have shown that various kinds of stem cells have the ability to recover premature ovarian aging (POA) function. Transplantation of human amniotic epithelial cells (hAECs) improves ovarian function damaged by chemotherapy in a mice model. Understanding of how to evaluate the distinct effects of adult stem cells in curing POA and how to choose stem cells in clinical application is lacking. To build a different degrees of POA model, mice were administered different doses of cyclophosphamide: light dose (70 mg/kg, 2 weeks), medium dose (70 mg/kg, 1 week; 120 mg/kg, 1 week), and high dose (120 mg/kg, 2 weeks). Enzyme-linked immunosorbent assay detected serum levels of sex hormones, and hematoxylin and eosin staining allowed follicle counting and showed the ovarian tissue structure. DiIC 18 (5)-DS was employed to label human amniotic mesenchymal stem cells (hAMSCs) and hAECs for detecting the cellular retention time in ovaries by a live imaging system. Proliferation of human ovarian granule cells (ki67, AMH, FSHR, FOXL2, and CYP19A1) and immunological rejection of human peripheral blood mononuclear cells (CD4, CD11b, CD19, and CD56) were measured by flow cytometry (fluorescence-activated cell sorting (FACS)). Distinction of cellular biological characteristics between hAECs and hAMSCs was evaluated, such as collagen secretory level (collagen I, II, III, IV, and VI), telomerase activity, pluripotent markers tested by western blot, expression level of immune molecules (HLA-ABC and HLA-DR) analyzed by FACS, and cytokines (growth factors, chemotactic factors, apoptosis factors, and inflammatory factors) measured by a protein antibody array methodology. After hAMSCs and hAECs were transplanted into a different degrees of POA model, hAMSCs exerted better therapeutic activity on mouse ovarian function in the high-dose administration group, promoting the proliferation rate of ovarian granular cells from premature ovarian failure patients, but also provoking immune

  4. Challenges in validating the sterilisation dose for processed human amniotic membranes

    International Nuclear Information System (INIS)

    Yusof, Norimah; Hassan, Asnah; Firdaus Abd Rahman, M.N.; Hamid, Suzina A.

    2007-01-01

    Most of the tissue banks in the Asia Pacific region have been using ionising radiation at 25 kGy to sterilise human tissues for save clinical usage. Under tissue banking quality system, any dose employed for sterilisation has to be validated and the validation exercise has to be a part of quality document. Tissue grafts, unlike medical items, are not produced in large number per each processing batch and tissues relatively have a different microbial population. A Code of Practice established by the International Atomic Energy Agency (IAEA) in 2004 offers several validation methods using smaller number of samples compared to ISO 11137 (1995), which is meant for medical products. The methods emphasise on bioburden determination, followed by sterility test on samples after they were exposed to verification dose for attaining of sterility assurance level (SAL) of 10 -1 . This paper describes our experience in using the IAEA Code of Practice in conducting the validation exercise for substantiating 25 kGy as sterilisation dose for both air-dried amnion and those preserved in 99% glycerol

  5. Challenges in validating the sterilisation dose for processed human amniotic membranes

    Energy Technology Data Exchange (ETDEWEB)

    Yusof, Norimah [Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia)], E-mail: norimah@mint.gov.my; Hassan, Asnah [Malaysian Nuclear Agency, Bangi, 43000 Kajang, Selangor (Malaysia); Firdaus Abd Rahman, M.N.; Hamid, Suzina A. [National Tissue Bank, Hospital Universiti Sains Malaysia, Kubang Kerian, 16130 Kelantan (Malaysia)

    2007-11-15

    Most of the tissue banks in the Asia Pacific region have been using ionising radiation at 25 kGy to sterilise human tissues for save clinical usage. Under tissue banking quality system, any dose employed for sterilisation has to be validated and the validation exercise has to be a part of quality document. Tissue grafts, unlike medical items, are not produced in large number per each processing batch and tissues relatively have a different microbial population. A Code of Practice established by the International Atomic Energy Agency (IAEA) in 2004 offers several validation methods using smaller number of samples compared to ISO 11137 (1995), which is meant for medical products. The methods emphasise on bioburden determination, followed by sterility test on samples after they were exposed to verification dose for attaining of sterility assurance level (SAL) of 10{sup -1}. This paper describes our experience in using the IAEA Code of Practice in conducting the validation exercise for substantiating 25 kGy as sterilisation dose for both air-dried amnion and those preserved in 99% glycerol.

  6. Decellularized human amniotic membrane: more is needed for an efficient dressing for protection of burns against antibiotic-resistant bacteria isolated from burn patients.

    Science.gov (United States)

    Gholipourmalekabadi, M; Bandehpour, M; Mozafari, M; Hashemi, A; Ghanbarian, H; Sameni, M; Salimi, M; Gholami, M; Samadikuchaksaraei, A

    2015-11-01

    Human amniotic membranes (HAMs) have attracted the attention of burn surgeons for decades due to favorable properties such as their antibacterial activity and promising support of cell proliferation. On the other hand, as a major implication in the health of burn patients, the prevalence of bacteria resistant to multiple antibiotics is increasing due to overuse of antibiotics. The aim of this study was to investigate whether HAMs (both fresh and acellular) are an effective antibacterial agent against antibiotic-resistant bacteria isolated from burn patients. Therefore, a HAM was decellularized and tested for its antibacterial activity. Decellularization of the tissue was confirmed by hematoxylin and eosin (H&E) and 4,6-diamidino-2-phenylindole (DAPI) staining. In addition, the cyto-biocompatibility of the acellular HAM was proven by the cell viability test (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide, MTT) and scanning electron microscopy (SEM). The resistant bacteria were isolated from burns, identified, and tested for their susceptibility to antibiotics using both the antibiogram and polymerase chain reaction (PCR) techniques. Among the isolated bacteria, three blaIMP gene-positive Pseudomonas aeruginosa strains were chosen for their high resistance to the tested antibiotics. The antibacterial activity of the HAM was also tested for Klebsiella pneumoniae (American Type Culture Collection (ATCC) 700603) as a resistant ATCC bacterium; Staphylococcus aureus (mecA positive); and three standard strains of ATCC bacteria including Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27833), and S. aureus (ATCC 25923). Antibacterial assay revealed that only the latter three bacteria were susceptible to the HAM. All the data obtained from this study suggest that an alternative strategy is required to complement HAM grafting in order to fully protect burns from nosocomial infections. Copyright © 2015 Elsevier Ltd and ISBI. All rights reserved.

  7. Amniotic Membrane Modifies the Genetic Program Induced by TGFß, Stimulating Keratinocyte Proliferation and Migration in Chronic Wounds.

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    Antonia Alcaraz

    Full Text Available Post-traumatic large-surface or deep wounds often cannot progress to reepithelialisation because they become irresponsive in the inflammatory stage, so intervention is necessary to provide the final sealing epidermis. Previously we have shown that Amniotic Membrane (AM induced a robust epithelialisation in deep traumatic wounds.To better understand this phenomenon, we used keratinocytes to investigate the effect of AM on chronic wounds. Using keratinocytes, we saw that AM treatment is able to exert an attenuating effect upon Smad2 and Smad3 TGFß-induced phosphorylation while triggering the activation of several MAPK signalling pathways, including ERK and JNK1, 2. This also has a consequence for TGFß-induced regulation on cell cycle control key players CDK1A (p21 and CDK2B (p15. The study of a wider set of TGFß regulated genes showed that the effect of AM was not wide but very concrete for some genes. TGFß exerted a powerful cell cycle arrest; the presence of AM however prevented TGFß-induced cell cycle arrest. Moreover, AM induced a powerful cell migration response that correlates well with the expression of c-Jun protein at the border of the healing assay. Consistently, the treatment with AM of human chronic wounds induced a robust expression of c-Jun at the wound border.The effect of AM on the modulation of TGFß responses in keratinocytes that favours proliferation together with AM-induced keratinocyte migration is the perfect match that allows chronic wounds to move on from their non-healing state and progress into epithelialization. Our results may explain why the application of AM on chronic wounds is able to promote epithelialisation.

  8. Transvaginal cervical length and amniotic fluid index: can it predict delivery latency following preterm premature rupture of membranes?

    Science.gov (United States)

    Mehra, Suwan; Amon, Erol; Hopkins, Sarah; Gavard, Jeffrey A; Shyken, Jaye

    2015-03-01

    We sought to determine whether transvaginal cervical length (TVCL), amniotic fluid index (AFI), or a combination of both can predict delivery latency within 7 days in women presenting with preterm premature rupture of membranes (PPROM). This was a prospective observational study of TVCL measurements in 106 singleton pregnancies with PPROM between 23-33 weeks. Delivery latency was defined as the period (in days) from the initial TVCL after PPROM to delivery of the infant, with our primary outcome being delivery within 7 days of TVCL. The independent predictability of significant characteristics for delivery within 7 days was determined using multiple logistic regression. Sensitivity, specificity, and predictive values were used to examine whether the presence of a short TVCL, AFI, or a combination of both affected the risk of delivery within 7 days. Delivery within 7 days occurred in 51/106 (48%) of pregnancies. Median duration (interquartile range) from PPROM to delivery and TVCL to delivery was 8 days (4.0-16.0) and 8 days (3.0-15.0), respectively. Using multiple regression TVCL as a continuous variable (odds ratio, 0.65; 95% confidence interval, 0.44-0.97; P 7 days for TVCL >2 cm alone was 61%. This predictive value changed when analyzed in conjunction with an AFI ≤5 cm and >5 cm at 42% and 89%, respectively. A shorter TVCL and an AFI ≤5 cm independently predict delivery within 7 days in women presenting with PPROM. The combination of an AFI >5 cm and TVCL >2 cm greatly improved the potential to remain undelivered at 7 days following cervical length assessment. These findings may be helpful for counseling and optimizing maternal and neonatal care in women with PPROM. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Non-invasively collected amniotic fluid as a source of possible biomarkers for premature rupture of membranes investigated by proteomic approach.

    Science.gov (United States)

    Consonni, Sara; Mainini, Veronica; Pizzardi, Agnese; Gianazza, Erica; Chinello, Clizia; Locatelli, Anna; Magni, Fulvio

    2014-02-01

    Preterm delivery is one of the main causes of perinatal morbidity and mortality and it accounts for 75 % of perinatal mortality and more than half of the long-term morbidity. We applied a proteomic approach based on mass spectrometry (MS) for biomarkers discovery of preterm premature rupture of membranes (pPROM) by investigating amniotic fluid (AF) invasively and non-invasively collected. Amniotic fluid was obtained from vagina of women with pPROM (group 1), PROM at term (group 2) and by genetic amniocentesis (group 3). Pre-fractionated AF proteome was analyzed through matrix assisted laser desorption ionization-time of flight (MALDI-TOF) MS. The characterization of proteins/peptides of interest was obtained by high performance liquid chromatography-electrospray tandem MS. Three peptides overexpressed in pPROM and able to discriminate the groups 1 and 2 were detected. One peptide was identified as the fragment Gly452LAVPDGPLGLPPKPro466 of the protein KIAA1522, expressed by fetal brain and liver. This peptide was overexpressed in a patient of the group 3, completely asymptomatic at the time of the amniocentesis, who later developed pPROM. Amniotic fluid invasively and non-invasively collected can be analyzed by MALDI-TOF MS to obtain proteomic profiles. Proteomic analysis identified a peptide with promising diagnostic capability for pPROM.

  10. Membrana amniótica como alternativa de tratamiento en superficie ocular Amniotic membrane as a therapeutic option for the ocular surface

    Directory of Open Access Journals (Sweden)

    Keyly Fernández García

    2012-12-01

    Full Text Available Se realizó una revisión bibliográfica con el objetivo de conocer la utilidad de la membrana amniótica como alternativa de tratamiento en la superficie ocular. Son abordados tópicos como las diferentes formas de obtención, preparación y conservación de la misma, sus mecanismos de acción y aplicaciones. Se consultó una bibliografía que abarca un periodo de varios años para conocer los resultados publicados sobre el trasplante de membrana amniótica humana en la superficie ocular.A literature review was made to learn about the usefulness of the amniotic membrane as a therapeutic option for the ocular surface. The different ways for obtaining, preparing, and conserving this membrane, its mechanism of action and its applications were also addressed. Literature covering several years was reviewed in order to be acquainted with the published results of the human amniotic membrane transplantation on the ocular surface.

  11. The use of amniotic membrane in the repair of duodenal wounds in Wistar rats Uso da membrana amniótica no reparo de feridas duodenais em ratos Wistar

    Directory of Open Access Journals (Sweden)

    Luciano Rodrigues Schimidt

    2010-02-01

    Full Text Available Purpose: In the search of a new material to repair duodenal wounds, a trial was conducted to assess the behavior of human amniotic membrane in the repair of the duodenal wall in rats. METHODS: Fifty Wistar rats weighing between 250 and 350g, male, were submitted to duodenotomy and randomly distributed into two groups. Group A (n=8 had no treatment and was used as the control group. In Group B (n=42 the duodenal wound was treated with a patch of human amniotic membrane. RESULTS: All animals in Group A died. In Group B no changes were observed with regards to death or the formation of duodenal fistula. All animals presented peritoneal adherences in the region on the duodenal wall repair and intestinal obstruction was observed in two animals. Healing of the duodenal wall in the region of the patch took place progressively as the post-operatory period increased, with regeneration of the mucosa and of the smooth muscle layer. CONCLUSION: From the clinical standpoint, the amniotic membrane proved to be a biological tissue which served as a temporary seal and allowed the wound to heal by second-intention, with re-establishment of the duodenal wall structure.OBJETIVO: Na busca de um novo material para o reparo dos ferimentos duodenais, foi efetuado um estudo para avaliar o comportamento da membrana amniótica humana no reparo da parede duodenal em ratos. MÉTODOS: Foram utilizados 50 ratos Wistar, com peso entre 250 e 350g, machos, distribuídos, aleatoriamente, em dois grupos. Grupo A (n=8, submetido à duodenotomia sem tratamento, utilizados como controle. Grupo B (n=42, submetido a um remendo de membrana amniótica humana para tratamento de ferimento duodenal provocado. RESULTADOS: Todos os animais do grupo A foram a óbito. No grupo B não foram observadas alterações quanto a óbito ou formação de fístula duodenal. Observaram-se em todos os animais aderências peritoneais à região do reparo da parede do duodeno e obstrução intestinal em

  12. Women's expectations and experiences of rupture of membranes and views of the potential use of reagent pads for detecting amniotic fluid.

    Science.gov (United States)

    Spiby, Helen; Borrelli, Sara; Hughes, Anita J

    2017-12-01

    To explore first-time mothers' expectations and experiences regarding rupture of membranes at term and their views on the potential use of reagent pads that detect amniotic fluid. There is little information available on women's experiences of spontaneous rupture of membranes, or interest in using methods to confirm rupture of membranes (e.g. reagent pads). Descriptive qualitative study, using focus groups and telephone interviews with women during pregnancy and after the birth of their first baby. Thematic analysis was undertaken to analyse women's responses. Ethics committee approval was obtained. Twenty-five women participated in the study of whom 13 contributed both during pregnancy and postpartum between October 2015-March 2016. Three overarching themes were identified from the data from women's expectations and experiences: uncertainty in how, when and where membranes may rupture; information which was felt to be limited and confirmation of rupture of membranes. The potential use of reagent pads met with varied responses. Women were interested in having facts and figures regarding rupture of membranes, such as characteristics of liquor; volume and probability of membranes rupturing spontaneously at term. Use of a pad as a means of confirmation was viewed as helpful, although the potential for increasing anxiety was raised. © 2017 John Wiley & Sons Ltd.

  13. ROCK1-directed basement membrane positioning coordinates epithelial tissue polarity.

    Science.gov (United States)

    Daley, William P; Gervais, Elise M; Centanni, Samuel W; Gulfo, Kathryn M; Nelson, Deirdre A; Larsen, Melinda

    2012-01-01

    The basement membrane is crucial for epithelial tissue organization and function. However, the mechanisms by which basement membrane is restricted to the basal periphery of epithelial tissues and the basement membrane-mediated signals that regulate coordinated tissue organization are not well defined. Here, we report that Rho kinase (ROCK) controls coordinated tissue organization by restricting basement membrane to the epithelial basal periphery in developing mouse submandibular salivary glands, and that ROCK inhibition results in accumulation of ectopic basement membrane throughout the epithelial compartment. ROCK-regulated restriction of PAR-1b (MARK2) localization in the outer basal epithelial cell layer is required for basement membrane positioning at the tissue periphery. PAR-1b is specifically required for basement membrane deposition, as inhibition of PAR-1b kinase activity prevents basement membrane deposition and disrupts overall tissue organization, and suppression of PAR-1b together with ROCK inhibition prevents interior accumulations of basement membrane. Conversely, ectopic overexpression of wild-type PAR-1b results in ectopic interior basement membrane deposition. Significantly, culture of salivary epithelial cells on exogenous basement membrane rescues epithelial organization in the presence of ROCK1 or PAR-1b inhibition, and this basement membrane-mediated rescue requires functional integrin β1 to maintain epithelial cell-cell adhesions. Taken together, these studies indicate that ROCK1/PAR-1b-dependent regulation of basement membrane placement is required for the coordination of tissue polarity and the elaboration of tissue structure in the developing submandibular salivary gland.

  14. Combined HLA matched limbal stem cells allograft with amniotic membrane transplantation as a prophylactic surgical procedure to prevent corneal graft rejection after penetrating keratoplasty: case report

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    Paolo Capozzi

    2014-09-01

    Full Text Available Purpose. To determine if the use of combined HLA matched limbal stem cells allograft with amniotic membrane transplantation (AMT is a safe and effective prophylactic surgical procedure to prevent corneal graft after penetrating keratoplasty (PK. Methods. We report the case of a 17 years old patient with a history of congenital glaucoma, trabeculectomy and multiple corneal graft rejections, presenting total limbal cell deficiency. To reduce the possibility of graft rejection in the left eye after a new PK, a two step procedure was performed. At first the patient underwent a combined HLA matched limbal stem cells allograft (LAT and AMT and then, 10 months later, a new PK. Results. During 12 months of follow-up, the corneal graft remained stable and smooth, with no sign of graft rejection. Conclusions. In our patient, the prophylactic use of LAT from HLA-matched donors and AMT before PK, may result in a better prognosis of corneal graft survival.

  15. An update clinical application of amniotic fluid-derived stem cells (AFSCs) in cancer cell therapy and tissue engineering.

    Science.gov (United States)

    Gholizadeh-Ghaleh Aziz, Shiva; Fathi, Ezzatollah; Rahmati-Yamchi, Mohammad; Akbarzadeh, Abolfazl; Fardyazar, Zahra; Pashaiasl, Maryam

    2017-06-01

    Recent studies have elucidated that cell-based therapies are promising for cancer treatments. The human amniotic fluid stem (AFS) cells are advantageous cells for such therapeutic schemes that can be innately changed to express therapeutic proteins. HAFSCs display a natural tropism to cancer cells in vivo. They can be useful in cancer cells targeting. Moreover, they are easily available from surplus diagnostic samples during pregnancy and less ethical and legal concern are associated with the collection and application than other putative cells are subjected. This review will designate representatives of amniotic fluid and stem cell derived from amniotic fluid. For this propose, we collect state of human AFS cells data applicable in cancer therapy by dividing this approach into two main classes (nonengineered and engineered based approaches). Our study shows the advantage of AFS cells over other putative cells types in terms differentiation ability to a wide range of cells by potential and effective use in preclinical studies for a variety of diseases. This study has shown the elasticity of human AFS cells and their favorable potential as a multipotent cell source for regenerative stem cell therapy and capable of giving rise to multiple lineages including such as osteoblasts and adipocyte.

  16. Identification of emergent motion compartments in the amniote embryo.

    Science.gov (United States)

    Loganathan, Rajprasad; Little, Charles D; Joshi, Pranav; Filla, Michael B; Cheuvront, Tracey J; Lansford, Rusty; Rongish, Brenda J

    2014-01-01

    The tissue scale deformations (≥ 1 mm) required to form an amniote embryo are poorly understood. Here, we studied ∼400 μm-sized explant units from gastrulating quail embryos. The explants deformed in a reproducible manner when grown using a novel vitelline membrane-based culture method. Time-lapse recordings of latent embryonic motion patterns were analyzed after disk-shaped tissue explants were excised from three specific regions near the primitive streak: 1) anterolateral epiblast, 2) posterolateral epiblast, and 3) the avian organizer (Hensen's node). The explants were cultured for 8 hours-an interval equivalent to gastrulation. Both the anterolateral and the posterolateral epiblastic explants engaged in concentric radial/centrifugal tissue expansion. In sharp contrast, Hensen's node explants displayed Cartesian-like, elongated, bipolar deformations-a pattern reminiscent of axis elongation. Time-lapse analysis of explant tissue motion patterns indicated that both cellular motility and extracellular matrix fiber (tissue) remodeling take place during the observed morphogenetic deformations. As expected, treatment of tissue explants with a selective Rho-Kinase (p160ROCK) signaling inhibitor, Y27632, completely arrested all morphogenetic movements. Microsurgical experiments revealed that lateral epiblastic tissue was dispensable for the generation of an elongated midline axis- provided that an intact organizer (node) is present. Our computational analyses suggest the possibility of delineating tissue-scale morphogenetic movements at anatomically discrete locations in the embryo. Further, tissue deformation patterns, as well as the mechanical state of the tissue, require normal actomyosin function. We conclude that amniote embryos contain tissue-scale, regionalized morphogenetic motion generators, which can be assessed using our novel computational time-lapse imaging approach. These data and future studies-using explants excised from overlapping anatomical

  17. The amniotic band syndrome: antenatal sonographic diagnosis and potential pitfalls.

    Science.gov (United States)

    Mahony, B S; Filly, R A; Callen, P W; Golbus, M S

    1985-05-01

    Amniotic band syndrome causes a variety of fetal malformations involving the limbs, craniofacial region, and trunk. Six prenatally diagnosed cases of amniotic band syndrome are discussed. The diagnosis was based on sonographic visualization of either amniotic sheets or bands associated with fetal deformation or deformities in nonembryologic distributions known to characterize the amniotic band syndrome. Seven additional cases are considered in which an aberrant sheet of tissue with a free edge was visualized within the amniotic cavity but no restriction of fetal motion or subsequent deformity was demonstrated.

  18. Self-assembly of tissue spheroids on polymeric membranes.

    Science.gov (United States)

    Messina, Antonietta; Morelli, Sabrina; Forgacs, Gabor; Barbieri, Giuseppe; Drioli, Enrico; De Bartolo, Loredana

    2017-07-01

    In this study, multicellular tissue spheroids were fabricated on polymeric membranes in order to accelerate the fusion process and tissue formation. To this purpose, tissue spheroids composed of three different cell types, myoblasts, fibroblasts and neural cells, were formed and cultured on agarose and membranes of polycaprolactone (PCL) and chitosan (CHT). Membranes prepared by a phase-inversion technique display different physicochemical, mechanical and transport properties, which can affect the fusion process. The membranes accelerated the fusion process of a pair of spheroids with respect to the inert substrate. In this process, a critical role is played by the membrane properties, especially by their mechanical characteristics and oxygen and carbon dioxide mass transfer. The rate of fusion was quantified and found to be similar for fibroblast, myoblast and neural tissue spheroids on membranes, which completed the fusion within 3 days. These spheroids underwent faster fusion and maturation on PCL membrane than on agarose, the rate of fusion being proportional to the value of oxygen and carbon dioxide permeances and elastic characteristics. Consequently, tissue spheroids on the membranes expressed high biological activity in terms of oxygen uptake, making them more suitable as building blocks in the fabrication of tissues and organs. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Membrane supported scaffold architectures for tissue engineering

    NARCIS (Netherlands)

    Bettahalli Narasimha, M.S.

    2011-01-01

    Tissue engineering aims at restoring or regenerating a damaged tissue. Often the tissue recreation occurs by combining cells, derived from a patient biopsy, onto a 3D porous matrix, functioning as a scaffold. One of the current limitations of tissue engineering is the inability to provide sufficient

  20. Hair Follicle Morphogenesis in the Treatment of Mouse Full-Thickness Skin Defects Using Composite Human Acellular Amniotic Membrane and Adipose Derived Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Wu Minjuan

    2016-01-01

    Full Text Available Early repair of skin injury and maximal restoration of the function and appearance have become important targets of clinical treatment. In the present study, we observed the healing process of skin defects in nude mice and structural characteristics of the new skin after transplantation of isolated and cultured adipose derived mesenchymal stem cells (ADMSCs onto the human acellular amniotic membrane (AAM. The result showed that ADMSCs were closely attached to the surface of AAM and grew well 24 h after seeding. Comparison of the wound healing rate at days 7, 14, and 28 after transplantation showed that ADMSCs seeded on AAM facilitated the healing of full-thickness skin wounds more effectively as compared with either hAM or AAM alone, indicating that ADMSCs participated in skin regeneration. More importantly, we noticed a phenomenon of hair follicle development during the process of skin repair. Composite ADMSCs and AAM not only promoted the healing of the mouse full-thickness defects but also facilitated generation of the appendages of the affected skin, thus promoting restoration of the skin function. Our results provide a new possible therapy idea for the treatment of skin wounds with respect to both anatomical regeneration and functional restoration.

  1. Membrana amniótica preservada em glicerina no reparo de feridas cutâneas de membros locomotores de eqüinos Equine amniotic membrane preserved in glycerin in reparing the equine limbs cütaneous wounds

    Directory of Open Access Journals (Sweden)

    Valdemir Alves de Oliveira

    1998-12-01

    treated with amniotic membrane, and the other, the control group, treated only with a damp gauze changed every 48 hours. In all the wounds the procedure was to evaluate the measurement of the área, degree of secretion, development of exuberant granulation tissue and the complete time of epitheliazation. Bacteriologic and histopatologic exams were made in a sample of the treated and contrai groups. The biochemical determination of proteins and the identification of fibroblastic growth factor were realized in the amnionic membrane. In all the treated wounds were observed shorter healing time, less formation of exuberant granulation tissue, smaller incidence of pathogenic bacterias and a smaller increase of the wound area.

  2. Effects of ionizing radiation on glycerolated amniotic membranes as a substract for cultured human epithelium; Efeitos da radiacao ionizante em membranas amnioticas gliceroladas empregadas como substrato ao cultivo de epitelio humano

    Energy Technology Data Exchange (ETDEWEB)

    Paggiaro, Andre Oliveira

    2011-07-01

    The amniotic membrane (AM) is a biomaterial with biological properties that are beneficial to tissue repair. It has been used as a temporary coverage to threat burns and chronic wounds. Recently, it has been served as a substrate for keratinocytes culture to construct a living skin equivalent. However, MA is a biological material, and its transplantation could cause infectious disease for receptors. So, it must be preserved and sterilized before clinical use. The aim of this study was to evaluate the radiation effects on glycerol-preserved MA, considering its compatibility to support human keratinocytes culture. Four MA were stored in high concentrations of glycerol (> 85%) and half of them were radio sterilized with a dose of 25 kGy. Then, we established two groups: nonirradiated MA (MA-ni) and irradiated MA (MA-i). Both groups was deepithelialized by a standardized protocol and was investigated morphologically, immunohistochemical and ultrastructural. Subsequently human keratinocytes were cultivated immersed and in air-liquid interface on denuded surface of MA-i and MA-ni. The results were compared at 14 and 21 days of culture by light and electron microscopy. After epithelial denudation, analyses demonstrated the continuity of the basement membrane in MA-ni group, whereas in the irradiated group, there was no indication of the basement membrane’s presence on the surface of MA. The cell cultures showed that in the non-irradiated group, there was growth of a multi-layered and differentiated epithelium, with a stratum corneum’s formation in air-liquid interface. In the irradiated group, the epithelium had only two or three layer, little cell differentiation, with the same results immersed or air-liquid interface system. Glycerol-preserved MA was biocompatible with the growth of a cultivated epithelium, showing its potential as a skin substitute. Irradiation at 25 kGy cause structural damage to the tissue, making changes in basement membrane, that facilitates

  3. Basement membrane chondroitin sulfate proteoglycans: localization in adult rat tissues

    DEFF Research Database (Denmark)

    McCarthy, K J; Couchman, J R

    1990-01-01

    Heparan sulfate proteoglycans have been described as the major proteoglycan component of basement membranes. However, previous investigators have also provided evidence for the presence of chondroitin sulfate glycosaminoglycan in these structures. Recently we described the production...... and characterization of core protein-specific monoclonal antibodies (MAb) against a chondroitin sulfate proteoglycan (CSPG) present in Reichert's membrane, a transient extra-embryonic structure of rodents. This CSPG was also demonstrated to be present in adult rat kidney. We report here the tissue distribution...... of epitopes recognized by these MAb. The ubiquitous presence of these epitopes in the basement membranes of nearly all adult rat tissues demonstrates that at least one CSPG is a constituent of most basement membranes, and by virtue of its unique distribution is distinct from other chondroitin and dermatan...

  4. Inflammatory Response of Human Gestational Membranes to Ureaplasma parvum Using a Novel Dual-Chamber Tissue Explant System.

    Science.gov (United States)

    Potts, Lauren C; Feng, Liping; Seed, Patrick C; Jayes, Friederike L; Kuchibhatla, Maragatha; Antczak, Brian; Nazzal, Matthew K; Murtha, Amy P

    2016-05-01

    Preterm premature rupture of membranes (PPROM) is often associated with intra-amniotic inflammation and infection. Current understanding of the pathogenesis of PPROM includes activation of pro-inflammatory cytokines and proteolytic enzymes leading to compromise of membrane integrity. The impact of exposure to bacterial pathogens, including Ureaplasma parvum, on gestational membranes is poorly understood. Our objective was to develop a dual-chamber system to characterize the inflammatory response of gestational membranes to U. parvum in a directional nature. Full-thickness human gestational membrane explants, with either choriodecidua or amnion oriented superiorly, were suspended between two washers in a cylindrical device, creating two distinct compartments. Brilliant green dye was introduced into the top chamber to assess the integrity of the system. Tissue viability was evaluated after 72 h using a colorimetric cell proliferation assay. Choriodecidua or amnion was exposed to three doses of U. parvum and incubated for 24 h. Following treatment, media from each compartment were used for quantification of U. parvum (quantitative PCR), interleukin (IL)-8 (enzyme-linked immunosorbent assay), and matrix metalloproteinase (MMP)-2 and MMP-9 activity (zymography). We observed that system integrity and explant viability were maintained over 72 h. Dose-dependent increases in recovered U. parvum, IL-8 concentration, and MMP-2 activity were detected in both compartments. Significant differences in IL-8 concentration and MMP-9 activity were found between the choriodecidua and amnion. This tissue explant system can be used to investigate the inflammatory consequences of directional bacterial exposure for gestational membranes and provides insight into the pathogenesis of PPROM and infectious complications of pregnancy. © 2016 by the Society for the Study of Reproduction, Inc.

  5. Affinity Labeling of Membrane Receptors Using Tissue-Penetrating Radiations

    Directory of Open Access Journals (Sweden)

    Franklin C. Wong

    2013-01-01

    Full Text Available Photoaffinity labeling, a useful in vivo biochemical tool, is limited when applied in vivo because of the poor tissue penetration by ultraviolet (UV photons. This study investigates affinity labeling using tissue-penetrating radiation to overcome the tissue attenuation and irreversibly label membrane receptor proteins. Using X-ray (115 kVp at low doses (<50 cGy or Rad, specific and irreversible binding was found on striatal dopamine transporters with 3 photoaffinity ligands for dopamine transporters, to different extents. Upon X-ray exposure (115 kVp, RTI-38 and RTI-78 ligands showed irreversible and specific binding to the dopamine transporter similar to those seen with UV exposure under other conditions. Similarly, gamma rays at higher energy (662 keV also affect irreversible binding of photoreactive ligands to peripheral benzodiazepine receptors (by PK14105 and to the dopamine (D2 membrane receptors (by azidoclebopride, respectively. This study reports that X-ray and gamma rays induced affinity labeling of membrane receptors in a manner similar to UV with photoreactive ligands of the dopamine transporter, D2 dopamine receptor (D2R, and peripheral benzodiazepine receptor (PBDZR. It may provide specific noninvasive irreversible block or stimulation of a receptor using tissue-penetrating radiation targeting selected anatomic sites.

  6. Amniotic fluid inflammatory cytokines

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Larsen, Nanna; Grove, Jakob

    2013-01-01

    The aim of the study was to analyze cytokine profiles in amniotic fluid (AF) samples of children developing autism spectrum disorders (ASD) and controls, adjusting for maternal autoimmune disorders and maternal infections during pregnancy.......The aim of the study was to analyze cytokine profiles in amniotic fluid (AF) samples of children developing autism spectrum disorders (ASD) and controls, adjusting for maternal autoimmune disorders and maternal infections during pregnancy....

  7. Autoinjerto conjuntival y membrana amniótica en la cirugía del pterigión primario Use of conjunctival autograft and amniotic membrane in primary pterygium surgery

    Directory of Open Access Journals (Sweden)

    Keyly Fernández García

    2012-12-01

    Full Text Available Objetivo: comparar la efectividad del autoinjerto conjuntival y el injerto de membrana amniótica en la cirugía del pterigión primario en el Instituto Cubano de Oftalmología "Ramón Pando Ferrer" de septiembre de 2009 a septiembre de 2010. Métodos: estudio prospectivo, comparativo y aleatorio. El universo estuvo constituido por 80 pacientes (80 ojos con diagnóstico de pterigión primario que se les realizó escisión quirúrgica de la hiperplasia fibrovascular nasal primaria. Se dividieron en 2 grupos al azar simple. Al grupo A (40 ojos se le implantó injerto de membrana amniótica y al grupo B (40 ojos autoinjerto de conjuntiva. Resultados: en seis meses de seguimiento se presentaron cuatro recurrencias (10 % en el grupo A y solo dos casos en el grupo B (5 %. En ninguno de los pacientes hubo complicación. Fue más efectivo el autoinjerto conjuntival. Conclusiones: la recidiva fue más frecuente en la cirugía con membrana amniótica aunque no existió una diferencia significativa. Se consideran ambas técnicas efectivas en esta afección.Objective: to compare the effectiveness of the conjunctival autograft and the amniotic membrane graft in the primary pterygium surgery at “Ramón Pando Ferrer” Cuban Institute of Ophthalmology from September 2009 to September 2010. Methods: a prospective, random and comparative study. The universe was composed by 80 eyes from 80 patients diagnosed with primary pterygium. They had undergone the surgical excision of a primary nasal fibrovascular hyperplasia. They were divided in two single-randomized groups (A and B; the group A patients (40 eyes were implanted with amniotic membrane graft whereas those of group B (40 eyes were implanted with conjunctival autograft. Results: during the 6 month follow-up, 4 recurrences were reported (10 % in group A and only 2 in group B (5 %. No complications were presented in both groups. The conjunctival autograft was more effective. Conclusions: the relapse was more

  8. Processo de reparação de lesões da córnea e a membrana amniótica na oftalmologia Repair process of corneal damage and the amniotic membrane in ophthalmology

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    Kelly Cristine de Sousa Pontes

    2011-12-01

    Full Text Available Os eventos que fazem parte do processo de reparação de lesões da córnea ocorrem simultaneamente e envolvem proliferação, migração, diferenciação e apoptose celular, além da comunicação intercelular. Vários fatores solúveis, além de proteínas da matriz mesenquimal, proteoglicanos, enzimas proteolíticas e alguns tipos celulares são abordados nesta revisão, na qual explicam-se os processos de reparação de lesões superficiais ou penetrantes da córnea. A membrana amniótica, muito utilizada na cirurgia oftálmica, foi estudada por apresentar funções que colaboram com o processo de reparação. Entretanto, tais funções poderão ser perdidas quando tal tecido for submetido à conservação. Assim, torna-se importante conhecer o processo de reparação de lesões que envolvem, ou não, a córnea em toda a sua espessura e escolher a melhor forma de utilização da membrana amniótica quando ela for indicada na terapia para estas lesões.The events included in the process of repair of corneal damage occur simultaneously and involve proliferation, migration, differentiation, cell apoptosis and intercellular communication. Several soluble factors, mesenchymal matrix proteins, proteoglycans, proteolytic enzymes and some cell types are covered in this review, which explains the processes of repair of corneal wounds, either superficial or penetrating. The amniotic membrane, used in ophthalmic surgery, was studied because of the contribution of its functions to the repair process. However, these functions may be lost when the amniotic membrane is subjected to conservation. Therefore, it is important to understand the repair process of lesions involving or not the entire thickness of the cornea, and choose the best use of the amniotic membrane, when it is indicated for the treatment of these lesions.

  9. Fetal- and uterine-specific antigens in human amniotic fluid.

    Science.gov (United States)

    Sutcliffe, R G; Brock, D J; Nicholson, L V; Dunn, E

    1978-09-01

    Removal of the major maternal serum proteins from second trimester amniotic fluid by antibody affinity chromatography revealed various soluble tissue antigens, of which two were fetal-specific skin proteins and another, of alpha2-mobility, was specific to the uterus, and was therefore designated alpha-uterine protein (AUP). These proteins could not be detected in maternal serum by antibody-antigen crossed electrophoresis. The concentration of AUP in amniotic fluid reached a maximum between 10 and 20 weeks of gestation, suggesting that there is an influx of uterine protein into the amniotic fluid at this stage of pregnancy.

  10. Timing of Histologic Progression from Chorio-Deciduitis to Chorio-Deciduo-Amnionitis in the Setting of Preterm Labor and Preterm Premature Rupture of Membranes with Sterile Amniotic Fluid.

    Science.gov (United States)

    Park, Chan-Wook; Park, Joong Shin; Norwitz, Errol R; Moon, Kyung Chul; Jun, Jong Kwan; Yoon, Bo Hyun

    2015-01-01

    Histologic chorio-deciduitis and chorio-deciduo-amnionitis (amnionitis) in extra-placental membranes are known to represent the early and advanced stages of ascending intra-uterine infection. However, there are no data in humans about the time required for chorio-deciduitis to develop and for chorio-deciduitis without amnionitis to progress to chorio-deciduitis with amnionitis, and the effect of prolongation of pregnancy on the development of chorio-deciduitis and amnionitis in patients with preterm labor and intact membranes (PTL) and preterm premature rupture of membranes (preterm-PROM). We examined these issues in this study. The study population consisted of 289 women who delivered preterm (133 cases with PTL, and 156 cases with preterm-PROM) and who had sterile amniotic fluid (AF) defined as a negative AF culture and the absence of inflammation as evidenced by a matrix metalloproteinase-8 (MMP-8) level membranes (i.e., inflammation-free extra-placental membranes, choroi-deciduitis only, and chorio-deciduitis with amnionitis) in patients with PTL and preterm-PROM. Amniocentesis-to-delivery interval was longer in cases of chorio-deciduitis with amnionitis than in cases of chorio-deciduitis only in both PTL (median [interquartile-range (IQR)]; 645.4 [319.5] vs. 113.9 [526.9] hours; P = 0.005) and preterm-PROM (131.3 [135.4] vs. 95.2 [140.5] hours; Pmembranes. Moreover, prolongation of pregnancy is an independent predictor of the development of both chorio-deciduitis and amnionitis in cases of PTL with sterile AF.

  11. Amniotic constriction bands

    Science.gov (United States)

    ... Supplements Videos & Tools Español You Are Here: Home → Medical Encyclopedia → Amniotic band sequence URL of this page: //medlineplus.gov/ency/ ... birth. The baby should be delivered in a medical center that has specialists experienced in caring for babies ... or partial loss of function of a body part. Congenital bands affecting large parts of the body cause the ...

  12. Melatonin membrane receptors in peripheral tissues: Distribution and functions

    Science.gov (United States)

    Slominski, Radomir M.; Reiter, Russel J.; Schlabritz-Loutsevitch, Natalia; Ostrom, Rennolds S.; Slominski, Andrzej T.

    2012-01-01

    Many of melatonin’s actions are mediated through interaction with the G-protein coupled membrane bound melatonin receptors type 1 and type 2 (MT1 and MT2, respectively) or, indirectly with nuclear orphan receptors from the RORα/RZR family. Melatonin also binds to the quinone reductase II enzyme, previously defined the MT3 receptor. Melatonin receptors are widely distributed in the body; herein we summarize their expression and actions in non-neural tissues. Several controversies still exist regarding, for example, whether melatonin binds the RORα/RZR family. Studies of the peripheral distribution of melatonin receptors are important since they are attractive targets for immunomodulation, regulation of endocrine, reproductive and cardiovascular functions, modulation of skin pigmentation, hair growth, cancerogenesis, and aging. Melatonin receptor agonists and antagonists have an exciting future since they could define multiple mechanisms by which melatonin modulates the complexity of such a wide variety of physiological and pathological processes. PMID:22245784

  13. [Heterogeneity of costs and performance for penetrating eye injuries and suturing amniotic membranes under DRG conditions : Analysis of case constellations of the G-DRG C01B of the Regensburg University eye clinic].

    Science.gov (United States)

    Franz, D; Mrosek, M; Mrosek, S; Helbig, H; Framme, C

    2012-01-01

    Patients with penetrating eye injuries are a very heterogeneous group both medically and economically. Since 2009, treatment involving sutures for open eye injuries and cases requiring amniotic membrane transplantation (AMT) were allocated to DRG C01B of the German diagnosis-related group system. However, given the significant clinical differences between these treatments, an inhomogeneity of costs to performance is postulated. This analysis describes case allocation problems within the G-DRG C01B category and presents solutions. A retrospective analysis was conducted from the standardized G-DRG data of 277 patients with open eye injuries and AMT between 2007 and 2008, grouped under the 2008 G-DRG system version to the G-DRG C01Z category. This data was provided by the Department of Ophthalmology at the University Hospital Regensburg. Additionally case-based data of the following were supplemented: length of surgery, time of anesthesia and intensity of patient care. Fixed and variable costs were determined for surgery and other inpatient treatment. Finally, an analysis of the heterogeneity of costs within the G-DRG C01B of the G-DRG system 2009 was implemented. Inhomogeneity was evident within the G-DRG C01B of the G-DRG system 2009 for the two groups suture of open eye injuries and AMT concerning the parameters length of stay, proportion of high outliers and cost per case. Multiple surgeries during an inpatient stay lead to an extended length of stay and increasing costs, especially within the AMT group. Intensity of patient care and the consideration of patient comorbidity did not yield relevant differences. The quality of the G-DRG system is measured by its ability to obtain adequate funding for highly complex and heterogeneous cases. Specific modifications of the G-DRG structures could increase the appropriateness of case allocation for patients with open eye injuries within the G-DRG C01B of the German DRG system 2009. As a result of the present study, cases

  14. Transplante de membrana amniótica associado ao transplante de conjuntiva autólogo no tratamento do pterígio primário Amniotic membrane transplantation associated with conjunctival autograft for primary pterygium treatment

    Directory of Open Access Journals (Sweden)

    Alessandra Guerra Daros Castellano

    2005-10-01

    Full Text Available OBJETIVO: Determinar a eficácia do transplante de membrana amniótica associada ao transplante de conjuntiva autólogo no tratamento do pterígio primário. MÉTODOS: Analisou-se prospectivamente 26 pacientes (14 mulheres e 12 homens com idade entre 20 e 60 anos, atendidos no ambulatório de oftalmologia do Hospital Universitário Evangélico de Curitiba e que foram submetidos ao tratamento do pterígio primário com cirurgia de exérese do pterígio e uso de membrana amniótica associada ao transplante de conjuntiva autólogo, entre abril e novembro de 2001. Foram analisados: grau do pterígio, complicações pós-operatórias e recidivas. RESULTADOS: No pós-operatório 4 casos (15,38% tiveram complicações, nenhuma considerada séria, sendo a mais freqüente a retração do enxerto em 2 casos. No sexto mês após a cirurgia, 24 pacientes (92,3% obtiveram sucesso cirúrgico e 2 pacientes (7,69% tiveram recidiva corneana ou conjuntival. Dos pacientes que tiveram complicações, 50% sofreram recidiva do pterígio (PPURPOSE: To evaluate the effectiveness of amniotic membrane transplantation associated with conjunctival autograft in primary pterygium treatment. METHODS: We have analyzed, prospectively, 26 patients (14 women and 12 men, from 20 to 60 years old, examined at the Evangelic Hospital Ophthalmology Outpatient Clinic. They were submitted to primary pterygium treatment with pterygium excision surgery, amniotic membrane transplantation and conjunctival autograft, from April to November, 2001. The evaluated aspects were: degree of pterygium, postoperative complications and recurrence. RESULTS: Four cases (15,38% showed complications, none of them considered serious, conjunctival graft contraction in 2 cases being the most frequent. After a follow-up of 6 months, 24 patients (92,3% had a successful result and 2 patients (7,69% had corneal or conjunctival recurrence. Among the patients with complications 50% presented pterygium recurrence (P

  15. Multiscale fabrication of biomimetic scaffolds for tympanic membrane tissue engineering

    International Nuclear Information System (INIS)

    Mota, Carlos; Danti, Serena; D’Alessandro, Delfo; Trombi, Luisa; Ricci, Claudio; Berrettini, Stefano; Puppi, Dario; Dinucci, Dinuccio; Chiellini, Federica; Milazzo, Mario; Stefanini, Cesare; Moroni, Lorenzo

    2015-01-01

    The tympanic membrane (TM) is a thin tissue able to efficiently collect and transmit sound vibrations across the middle ear thanks to the particular orientation of its collagen fibers, radiate on one side and circular on the opposite side. Through the combination of advanced scaffolds and autologous cells, tissue engineering (TE) could offer valuable alternatives to autografting in major TM lesions. In this study, a multiscale approach based on electrospinning (ES) and additive manufacturing (AM) was investigated to fabricate scaffolds, based on FDA approved copolymers, resembling the anatomic features and collagen fiber arrangement of the human TM. A single scale TM scaffold was manufactured using a custom-made collector designed to confer a radial macro-arrangement to poly(lactic-co-glycolic acid) electrospun fibers during their deposition. Dual and triple scale scaffolds were fabricated combining conventional ES with AM to produce poly(ethylene oxide terephthalate)/poly(butylene terephthalate) block copolymer scaffolds with anatomic-like architecture. The processing parameters were optimized for each manufacturing method and copolymer. TM scaffolds were cultured in vitro with human mesenchymal stromal cells, which were viable, metabolically active and organized following the anisotropic character of the scaffolds. The highest viability, cell density and protein content were detected in dual and triple scale scaffolds. Our findings showed that these biomimetic micro-patterned substrates enabled cell disposal along architectural directions, thus appearing as promising substrates for developing functional TM replacements via TE. (paper)

  16. Patch esophagoplasty using an in-body-tissue-engineered collagenous connective tissue membrane.

    Science.gov (United States)

    Okuyama, Hiroomi; Umeda, Satoshi; Takama, Yuichi; Terasawa, Takeshi; Nakayama, Yasuhide

    2018-02-01

    Although many approaches to esophageal replacement have been investigated, these efforts have thus far only met limited success. In-body-tissue-engineered connective tissue tubes have been reported to be effective as vascular replacement grafts. The aim of this study was to investigate the usefulness of an In-body-tissue-engineered collagenous connective tissue membrane, "Biosheet", as a novel esophageal scaffold in a beagle model. We prepared Biosheets by embedding specially designed molds into subcutaneous pouches in beagles. After 1-2months, the molds, which were filled with ingrown connective tissues, were harvested. Rectangular-shaped Biosheets (10×20mm) were then implanted to replace defects of the same size that had been created in the cervical esophagus of the beagle. An endoscopic evaluation was performed at 4 and 12weeks after implantation. The esophagus was harvested and subjected to a histological evaluation at 4 (n=2) and 12weeks (n=2) after implantation. The animal study protocols were approved by the National Cerebral and Cardiovascular Centre Research Institute Committee (No. 16048). The Biosheets showed sufficient strength and flexibility to replace the esophagus defect. All animals survived with full oral feeding during the study period. No anastomotic leakage was observed. An endoscopic study at 4 and 12weeks after implantation revealed that the anastomotic sites and the internal surface of the Biosheets were smooth, without stenosis. A histological analysis at 4weeks after implantation demonstrated that stratified squamous epithelium was regenerated on the internal surface of the Biosheets. A histological analysis at 12weeks after implantation showed the regeneration of muscle tissue in the implanted Biosheets. The long-term results of patch esophagoplasty using Biosheets showed regeneration of stratified squamous epithelium and muscular tissues in the implanted sheets. These results suggest that Biosheets may be useful as a novel esophageal

  17. Liquid chromatography--tandem mass spectrometry analysis of cocaine and its metabolites from blood, amniotic fluid, placental and fetal tissues: study of the metabolism and distribution of cocaine in pregnant rats.

    Science.gov (United States)

    Srinivasan, K; Wang, P P; Eley, A T; White, C A; Bartlett, M G

    2000-08-18

    The ability to simultaneously quantitate cocaine and its 12 metabolites from pregnant rat blood, amniotic fluid, placental and fetal tissue homogenates aids in elucidating the metabolism and distribution of cocaine. An efficient extraction method was developed to simultaneously recover these 13 components using underivatized silica solid-phase extraction (SPE) cartridges. The overall recoveries for cocaine and its metabolites were studied from pregnant rat blood (47-100%), amniotic fluid (61-100%), placental homogenate (31-83%), and fetal homogenate (39-87%). Extraction of the samples using silica is not classical SPE, but rather allows for the concentration of the sample into a small volume prior to injection and the removal of the proteins due to their strong interaction with the active silica surface. A positive ion mode electrospray ionization liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was used and validated to simultaneously quantitate cocaine and 12 metabolites from these four biological matrices. A gradient elution method with a Zorbax XDB C8 reversed-phase column was used to separate the components. Multiple reaction monitoring (MRM) of a product ion arising from the corresponding precursor ion was used in order to enhance the selectivity and sensitivity of the method. Low background noise was observed from the complex biological matrices due to efficient SPE and the selectivity of the MRM mode. Linear calibration curves were generated from 0.01 to 2.50 ppm. The method also showed high intra-day (n =3) and inter-day (n=9) precision (% RSD) and accuracy (% error) for all components. The limits of detection (LODs) for the method ranged from 0.15 to 10 ppb. The LODs of cocaine and its major metabolites were less than 1 ppb from all four biological matrices. This method was applied to the study of the metabolism and distribution of cocaine in pregnant rats following intravenous infusion to a steady state plasma drug concentration. The

  18. Amniotic Fluid Cells Proliferation in Normal and Down Syndrome Subjects

    Directory of Open Access Journals (Sweden)

    Honcea Adina

    2016-02-01

    Full Text Available Down Syndrome/Trisomy 21 is the most common chromosomal anomaly, and it represents the most common congenital cause of infants’ intellectual disability. Subjects with this syndrome are affected by degenerative processes caused by accelerated aging or unknown ethyologies. In recent years, accumulating evidence revealed increased potential of amniotic fluid-derived stem cells to be used in regenerative therapy. Our aim was to assess differences in immunophenotype, cell morphology and proliferation of amniotic fluid cells from normal and Down Syndrome pregnancies using a quantitative cytometry approach. Results revealed the emergence of a population of small sized cells in Down Syndrome derived amniotic fluid cells that are readily visible upon microscopic inspection. Hence, the fluorescence–based quantitative image cytometry determinations showed a tendency of decrease in both cell and nuclei size in trisomy, with no significant modification in nuclei circularity, as measured following actin cytoskeleton and nuclei labeling. The propensity of Ki67 positive cells was found to be increased in Down Syndrome derived cells (48.92% as compared to normal specimens (28.68%. However, cells in S and G2/M cell cycle phases decreased from 32.91% to 4.49% in diseased cells. Further studies are devoted to understanding the molecular basis of the observed differences in the proliferation ability of Down Syndrome amniotic cells, in order to evaluate the potential therapeutic effect of amniotic fluid stem cells for tissue regeneration in subjects with trisomy and to find correlations between amniotic cells phenotype and patient prognosis.

  19. * Human Amniotic Mesenchymal Stromal Cells as Favorable Source for Cartilage Repair.

    Science.gov (United States)

    Muiños-López, Emma; Hermida-Gómez, Tamara; Fuentes-Boquete, Isaac; de Toro-Santos, Javier; Blanco, Francisco Javier; Díaz-Prado, Silvia María

    2017-09-01

    Localized trauma-derived breakdown of the hyaline articular cartilage may progress toward osteoarthritis, a degenerative condition characterized by total loss of articular cartilage and joint function. Tissue engineering technologies encompass several promising approaches with high therapeutic potential for the treatment of these focal defects. However, most of the research in tissue engineering is focused on potential materials and structural cues, while little attention is directed to the most appropriate source of cells endowing these materials. In this study, using human amniotic membrane (HAM) as scaffold, we defined a novel static in vitro model for cartilage repair. In combination with HAM, four different cell types, human chondrocytes, human bone marrow-derived mesenchymal stromal cells (hBMSCs), human amniotic epithelial cells, and human amniotic mesenchymal stromal cells (hAMSCs) were assessed determining their therapeutic potential. A chondral lesion was drilled in human cartilage biopsies simulating a focal defect. A pellet of different cell types was implanted inside the lesion and covered with HAM. The biopsies were maintained for 8 weeks in culture. Chondrogenic differentiation in the defect was analyzed by histology and immunohistochemistry. HAM scaffold showed good integration and adhesion to the native cartilage in all groups. Although all cell types showed the capacity of filling the focal defect, hBMSCs and hAMSCs demonstrated higher levels of new matrix synthesis. However, only the hAMSCs-containing group presented a significant cytoplasmic content of type II collagen when compared with chondrocytes. More collagen type I was identified in the new synthesized tissue of hBMSCs. In accordance, hBMSCs and hAMSCs showed better International Cartilage Research Society scoring although without statistical significance. HAM is a useful material for articular cartilage repair in vitro when used as scaffold. In combination with hAMSCs, HAM showed better

  20. Management of Amniotic Sheet with a Hammock-like Placenta

    Directory of Open Access Journals (Sweden)

    Liangcheng Wang

    2016-09-01

    Full Text Available An amniotic sheet is a septation in the amniotic cavity with a perforation that allows amniotic fluid to pass through. Although the incidence of abnormal placental implantation is higher in such cases, the management recommendations remain unclear. We report a case of an amniotic sheet with a hammock-like placenta located in the center of the uterine cavity. A 25-year-old woman with a history of two dilation and curettage procedures was found to have an amniotic cavity separated by a septum that contained part of the placenta. At gestational Week 32, magnetic resonance images revealed that the placenta was attached from the anterior to posterior uterine walls and resembled a hammock hanging in the center of the uterus. Subsequently, continuous intravenous administration of ritodrine hydrochloride and magnesium sulfate were given. The pregnancy was extended to Week 36. Elective cesarean section was performed, and a 3212-g female infant was delivered. Thus, owing to the risk of umbilical cord complications and placental injury secondary to premature rupture of membranes, aggressive and careful perinatal management is required in such cases.

  1. Perkembangan Terkini Membran Guided Tissue Regeneration/Guided Bone Regeneration sebagai Terapi Regenerasi Jaringan Periodontal

    Directory of Open Access Journals (Sweden)

    Cindy Cahaya

    2015-06-01

    kombinasi prosedur-prosedur di atas, termasuk prosedur bedah restoratif yang berhubungan dengan rehabilitasi oral dengan penempatan dental implan. Pada tingkat selular, regenerasi periodontal adalah proses kompleks yang membutuhkan proliferasi yang terorganisasi, differensiasi dan pengembangan berbagai tipe sel untuk membentuk perlekatan periodontal. Rasionalisasi penggunaan guided tissue regeneration sebagai membran pembatas adalah menahan epitel dan gingiva jaringan pendukung, sebagai barrier membrane mempertahankan ruang dan gigi serta menstabilkan bekuan darah. Pada makalah ini akan dibahas sekilas mengenai 1. Proses penyembuhan terapi periodontal meliputi regenerasi, repair ataupun pembentukan perlekatan baru. 2. Periodontal spesific tissue engineering. 3. Berbagai jenis membran/guided tissue regeneration yang beredar di pasaran dengan keuntungan dan kerugian sekaligus karakteristik masing-masing membran. 4. Perkembangan membran terbaru sebagai terapi regenerasi penyakit periodontal. Tujuan penulisan untuk memberi gambaran masa depan mengenai terapi regenerasi yang menjanjikan sebagai perkembangan terapi penyakit periodontal.   Latest Development of Guided Tissue Regeneration and Guided Bone Regeneration Membrane as Regenerative Therapy on Periodontal Tissue. Periodontitis is a patological state which influences the integrity of periodontal system that could lead to the destruction of the periodontal tissue and end up with tooth loss. Currently, there are so many researches and efforts to regenerate periodontal tissue, not only to stop the process of the disease but also to reconstruct the periodontal tissue. Periodontal regenerative therapy aims at directing the growth of new bone, cementum and periodontal ligament on the affected teeth. Regenerative procedures consist of soft tissue graft, bone graft, roots biomodification, guided tissue regeneration and combination of the procedures, including restorative surgical procedure that is

  2. Estudo por microscopia eletrônica do epitélio de neovaginas confeccionadas com membrana amniótica e pesquisa de receptores para estrogênios Electron microscope study of the epithelium of neovagina constructed with amniotic membrane: estrogen receptor evaluation

    Directory of Open Access Journals (Sweden)

    Mauri José Piazza

    1999-06-01

    cortes histológicos obtidos em biópsias de neovaginas revestidas com membrana amniótica.Purpose: to verify changes in amnion epithelia after neovaginoplasty and the presence of estrogen receptor in this tissue using electron microscopy (TEM. Methods: a group consisting of 33 young patients with vaginal agenesy, most of them amenorreic, either incapable or having difficulty to adequately perform sexual activity, underwent surgical correction of the anomaly by the McIndoe-Banister technique. In each woman, a cavity was created through an opening of the vesicorectal space. A mold made of artificial sponge with a condom recovered by amniotic membrane was introduced into the cavity. After eight days, the mold was removed, leaving a developing neoepithelium from the amniotic membrane. Results: The TEM conducted in samples of tissue obtained from neovaginas showed similar results in all three layers of these epithelia, either in the cytoplasm or the cellular microstructures in comparison with normal women. The intensity of staining of the estrogen receptors on the vaginal neoepithelium of the group of women who underwent the surgical procedure was similar to that of normal women. The estrogen receptors were heterogeneously distributed in the three layers of the tissue in all women. The analysis of the vaginal neoepithelium obtained from amniotic membrane revealed, through the TEM, all the characteristics of a regular vaginal epithelium.

  3. Medical applications of membranes: Drug delivery, artificial organs and tissue engineering

    NARCIS (Netherlands)

    Stamatialis, Dimitrios; Papenburg, B.J.; Girones nogue, Miriam; Saiful, S.; Bettahalli Narasimha, M.S.; Schmitmeier, Stephanie; Wessling, Matthias

    2008-01-01

    This paper covers the main medical applications of artificial membranes. Specific attention is given to drug delivery systems, artificial organs and tissue engineering which seem to dominate the interest of the membrane community this period. In all cases, the materials, methods and the current

  4. Neovaginoplastia com membrana amniótica na síndrome de Mayer-Rokitansky-Küster-Hauser Neovaginoplasty using amniotic membrane in Mayer-Rokitansky-Küster-Hauser syndrome

    Directory of Open Access Journals (Sweden)

    Bruno Ramalho de Carvalho

    2007-12-01

    neovagina de aspecto adequado.PURPOSE: to evaluate the results of neovaginoplasty with the use of a human amniotic graft in patients with the Mayer-Rokitansky-Küster-Hauser (MRKH syndrome. METHODS: the study was a retrospective analysis of a series of 28 patients with the MRKH syndrome conducted from 1990 to 2003. The patients were attended and treated at the Ambulatório de Ginecologia Infanto-Puberal (AGIP of the Hospital Universitário of the Faculdade de Medicina de Ribeirão Preto of the Universidade de São Paulo (FMRP-USP, being submitted to neovaginoplasty by the technique of McIndoe and Bannister, modified by the use of a human amniotic membrane graft. Epithelization, amplitude and depth of the neovaginas were evaluated 7 and 40 days after the procedure. Patient satisfaction was determined during the late postoperative period in terms of the presence of discomfort and dyspareunia during sexual relations. RESULTS: postoperatively, seven patients (25% presented vaginal stenosis and six of them were submitted to a new surgical intervention, one had shortening of the neovagina, corrected with the use of exercises with a vaginal mold, three (10.7% developed a rectovaginal fistula, one (3.6% a uterovesical fistula, and one (3.6% excess skin in the vaginal introitus - all successfully corrected with surgery. Four patients (14.3% presented urinary tract infection. Two months after surgery, 11/19 patients (57.8% presented satisfactory sexual activity and 42% dyspareunia, and within a maximum period of four years, 20/21 patients (95.2% had satisfactory sexual activity and 4.8% dyspareunia. CONCLUSIONS: an amniotic membrane graft is a good option for the treatment of vaginal agenesis. Perioperative follow-up involves educational guidance regarding the use of the mold and regarding patient sexuality in order to reduce the complaints of dysfunctional coitus in the presence of a favorable surgical evolution and a neovagina of adequate aspect.

  5. Protein adsorption capability on polyurethane and modified-polyurethane membrane for periodontal guided tissue regeneration applications

    Energy Technology Data Exchange (ETDEWEB)

    Sheikh, Zeeshan [Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Fitzgerald Building, 150 College Street, Toronto, ON M5S 3E2 (Canada); School of Engineering and Materials Science, Queen Mary, University of London, Mile End Rd, London, E1 4NS (United Kingdom); Khan, Abdul Samad, E-mail: draskhan@ciitlahore.edu.pk [Interdisciplinary Research Centre in Biomedical Materials, COMSATS Institute of Information Technology, Lahore 54000 (Pakistan); Roohpour, Nima [Oral Care R& D, GSK St., Georges Ave., Weybridge KT13 8PA (United Kingdom); Glogauer, Michael [Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Fitzgerald Building, 150 College Street, Toronto, ON M5S 3E2 (Canada); Rehman, Ihtesham u [Department of Materials Science and Engineering, The Kroto Research Institute, North Campus, University of Sheffield, Broad Lane, Sheffield S3 7HQ (United Kingdom)

    2016-11-01

    Periodontal disease if left untreated can result in creation of defects within the alveolar ridge. Barrier membranes are frequently used with or without bone replacement graft materials for achieving periodontal guided tissue regeneration (GTR). Surface properties of barrier membranes play a vital role in their functionality and clinical success. In this study polyetherurethane (PEU) membranes were synthesized by using 4,4′-methylene-diphenyl diisocyanate (MDI), polytetramethylene oxide (PTMO) and 1,4-butane diol (BDO) as a chain extender via solution polymerization. Hydroxyl terminated polydimethylsiloxane (PDMS) due to having inherent surface orientation towards air was used for surface modification of PEU on one side of the membranes. This resulting membranes had one surface being PEU and the other being PDMS coated PEU. The prepared membranes were treated with solutions of bovine serum albumin (BSA) in de-ionized water at 37 °C at a pH of 7.2. The surface protein adsorptive potential of PEU membranes was observed using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Raman spectroscopy and Confocal Raman spectroscopy. The contact angle measurement, tensile strength and modulus of prepared membranes were also evaluated. PEU membrane (89.86 ± 1.62°) exhibited less hydrophobic behavior than PEU-PDMS (105.87 ± 3.16°). The ultimate tensile strength and elastic modulus of PEU (27 ± 1 MPa and 14 ± 2 MPa) and PEU-PDMS (8 ± 1 MPa and 26 ± 1 MPa) membranes was in required range. The spectral analysis revealed adsorption of BSA proteins on the surface of non PDMS coated PEU surface. The PDMS modified PEU membranes demonstrated a lack of BSA adsorption. The non PDMS coated side of the membrane which adsorbs proteins could potentially be used facing towards the defect attracting growth factors for periodontal tissue regeneration. Whereas, the PDMS coated side could serve as an occlusive barrier for preventing gingival epithelial

  6. Protein adsorption capability on polyurethane and modified-polyurethane membrane for periodontal guided tissue regeneration applications

    International Nuclear Information System (INIS)

    Sheikh, Zeeshan; Khan, Abdul Samad; Roohpour, Nima; Glogauer, Michael; Rehman, Ihtesham u

    2016-01-01

    Periodontal disease if left untreated can result in creation of defects within the alveolar ridge. Barrier membranes are frequently used with or without bone replacement graft materials for achieving periodontal guided tissue regeneration (GTR). Surface properties of barrier membranes play a vital role in their functionality and clinical success. In this study polyetherurethane (PEU) membranes were synthesized by using 4,4′-methylene-diphenyl diisocyanate (MDI), polytetramethylene oxide (PTMO) and 1,4-butane diol (BDO) as a chain extender via solution polymerization. Hydroxyl terminated polydimethylsiloxane (PDMS) due to having inherent surface orientation towards air was used for surface modification of PEU on one side of the membranes. This resulting membranes had one surface being PEU and the other being PDMS coated PEU. The prepared membranes were treated with solutions of bovine serum albumin (BSA) in de-ionized water at 37 °C at a pH of 7.2. The surface protein adsorptive potential of PEU membranes was observed using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Raman spectroscopy and Confocal Raman spectroscopy. The contact angle measurement, tensile strength and modulus of prepared membranes were also evaluated. PEU membrane (89.86 ± 1.62°) exhibited less hydrophobic behavior than PEU-PDMS (105.87 ± 3.16°). The ultimate tensile strength and elastic modulus of PEU (27 ± 1 MPa and 14 ± 2 MPa) and PEU-PDMS (8 ± 1 MPa and 26 ± 1 MPa) membranes was in required range. The spectral analysis revealed adsorption of BSA proteins on the surface of non PDMS coated PEU surface. The PDMS modified PEU membranes demonstrated a lack of BSA adsorption. The non PDMS coated side of the membrane which adsorbs proteins could potentially be used facing towards the defect attracting growth factors for periodontal tissue regeneration. Whereas, the PDMS coated side could serve as an occlusive barrier for preventing gingival epithelial

  7. LHRH-pituitary plasma membrane binding: the presence of specific binding sites in other tissues.

    Science.gov (United States)

    Marshall, J C; Shakespear, R A; Odell, W D

    1976-11-01

    Two specific binding sites for LHRH are present on plasma membranes prepared from rat and bovine anterior pituitary glands. One site is of high affinity (K = 2X108 1/MOL) and the second is of lower affinity (8-5X105 1/mol) and much greater capacity. Studies on membrane fractions prepared from other tissues showed the presence of a single specific site for LHRH. The kinetics and specificity of this site were similar to those of the lower affinity pituitary receptor. These results indicate that only pituitary membranes possess the higher affinity binding site and suggest that the low affinity site is not of physiological importance in the regulation of gonadotrophin secretion. After dissociation from membranes of non-pituitary tissues 125I-LHRH rebound to pituitary membrane preparations. Thus receptor binding per se does not result in degradation of LHRH and the function of these peripheral receptors remains obscure.

  8. Mechanical properties of electrospun bilayer fibrous membranes as potential scaffolds for tissue engineering.

    Science.gov (United States)

    Pu, Juan; Komvopoulos, Kyriakos

    2014-06-01

    Bilayer fibrous membranes of poly(l-lactic acid) (PLLA) were fabricated by electrospinning, using a parallel-disk mandrel configuration that resulted in the sequential deposition of a layer with fibers aligned across the two parallel disks and a layer with randomly oriented fibers, both layers deposited in a single process step. Membrane structure and fiber alignment were characterized by scanning electron microscopy and two-dimensional fast Fourier transform. Because of the intricacies of the generated electric field, bilayer membranes exhibited higher porosity than single-layer membranes consisting of randomly oriented fibers fabricated with a solid-drum collector. However, despite their higher porosity, bilayer membranes demonstrated generally higher elastic modulus, yield strength and toughness than single-layer membranes with random fibers. Bilayer membrane deformation at relatively high strain rates comprised multiple abrupt microfracture events characterized by discontinuous fiber breakage. Bilayer membrane elongation yielded excessive necking of the layer with random fibers and remarkable fiber stretching (on the order of 400%) in the layer with fibers aligned in the stress direction. In addition, fibers in both layers exhibited multiple localized necking, attributed to the nonuniform distribution of crystalline phases in the fibrillar structure. The high membrane porosity, good mechanical properties, and good biocompatibility and biodegradability of PLLA (demonstrated in previous studies) make the present bilayer membranes good scaffold candidates for a wide range of tissue engineering applications. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  9. The efficacy of topical human amniotic membrane-mesenchymal stem cell-conditioned medium (hAMMSC-CM) and a mixture of topical hAMMSC-CM + vitamin C and hAMMSC-CM + vitamin E on chronic plantar ulcers in leprosy:a randomized control trial.

    Science.gov (United States)

    Prakoeswa, C R S; Natallya, F R; Harnindya, D; Thohiroh, A; Oktaviyanti, R N; Pratiwi, K D; Rubianti, M A; Yogatri, B; Primasari, P I; Herwanto, N; Alinda, M D; Kusumaputra, B H; Astari, L; Listiawan, M Y; Agusni, I; Rantam, F A

    2018-05-10

    Healing of chronic plantar ulcers in leprosy (CPUL) typically takes a long time due to impaired neurological function, thereby reducing the levels of growth factors and cytokines. Cytokines can be found in metabolite products from amniotic membrane stem cells. Chronic ulcers are frequently characterized by high levels of reactive oxygen species. Vitamin E (α-tocopherol) is widely used in skin lesions, owing to its antioxidant and anti-inflammatory properties. Vitamin C also has antioxidant, anti-inflammatory, and collagen synthesis properties which are useful in wound healing. Herein, we compared the effects of topical human amniotic membrane-mesenchymal stem cell-conditioned medium (hAMMSC-CM) alone and with vitamins C and E on healing of CPUL. In this randomized controlled trial, topical agents were applied every 3 days for up to 8 weeks. Ulcer size, side-effects, and possible complications were monitored weekly. Healing percentage increased each week in all groups. Mean difference in ulcer size was highest in the hAMMSC-CM + vitamin E group, implying better progress of wound healing. There were no side-effects or complications. hAMMSC-CM + vitamin E is best for healing of CPUL.

  10. Human amniotic epithelial cells combined with silk fibroin scaffold in the repair of spinal cord injury

    Directory of Open Access Journals (Sweden)

    Ting-gang Wang

    2016-01-01

    Full Text Available Treatment and functional reconstruction after central nervous system injury is a major medical and social challenge. An increasing number of researchers are attempting to use neural stem cells combined with artificial scaffold materials, such as fibroin, for nerve repair. However, such approaches are challenged by ethical and practical issues. Amniotic tissue, a clinical waste product, is abundant, and amniotic epithelial cells are pluripotent, have low immunogenicity, and are not the subject of ethical debate. We hypothesized that amniotic epithelial cells combined with silk fibroin scaffolds would be conducive to the repair of spinal cord injury. To test this, we isolated and cultured amniotic epithelial cells, and constructed complexes of these cells and silk fibroin scaffolds. Implantation of the cell-scaffold complex into a rat model of spinal cord injury resulted in a smaller glial scar in the damaged cord tissue than in model rats that received a blank scaffold, or amniotic epithelial cells alone. In addition to a milder local immunological reaction, the rats showed less inflammatory cell infiltration at the transplant site, milder host-versus-graft reaction, and a marked improvement in motor function. These findings confirm that the transplantation of amniotic epithelial cells combined with silk fibroin scaffold can promote the repair of spinal cord injury. Silk fibroin scaffold can provide a good nerve regeneration microenvironment for amniotic epithelial cells.

  11. RESEARCH ON REDUCING PREMATURITY RUPTURE OF MEMBRANE

    OpenAIRE

    Maria URSACHI (BOLOTA); Emil ANTON; Sorana Caterina ANTON

    2016-01-01

    The membranes surrounding the amniotic cavity are composed from amnion and chorion, tightly adherent layers which are composed of several cell types, including epithelial cells, trophoblasts cells and mesenchyme cells, embedded in a collagenous matrix. They retain amniotic fluid, secret substances into the amniotic fluid, as well as to the uterus and protect the fetus against upward infections from urogenital tract. Normally, the membranes it breaks during labor. Premature rupture of the amn...

  12. The properties of connective tissue membrane and pig skin as raw materials for cooked sausage.

    Science.gov (United States)

    Puolanne, E; Ruusunen, M

    1981-09-01

    Pig skin and epimysial membrane from young bulls were comminuted in a colloid grinder and mixed with water and additives. The resultant mixture was heated in a water bath to give an internal temperature of 72°C, and centrifuged while still hot. Such variations in the amount of water added, the salt content, the phosphate content and the pH value as are possible in cooked sausage heated to over 65°C during processing did not cause marked changes in the amount of water bound by the connective tissues, the amount of dissolved protein or the gel strength of the liquid released from the connective tissues. As the temperature rose the amount of bound water dropped, but the amount of dissolved protein and the gel strength increased. The liquid released from the connective tissue membranes formed a gel at 32°C and re-melted at 49°C. For pig skin, the corresponding temperatures were 23°C and 47°C. On the basis of this study it appears that connective tissue may be important for the water-binding capacity and firmness of cold sausage. The connective tissue membranes obtained from young bulls and pig skin are of roughly equal value in this respect, although the gel formed from connective tissue membrane is tougher. Copyright © 1981. Published by Elsevier Ltd.

  13. In vitro evaluation of electrospun chitosan mats crosslinked with genipin as guided tissue regeneration barrier membranes

    Science.gov (United States)

    Norowski, Peter Andrew, Jr.

    Guided tissue regeneration (GTR) is a surgical technique commonly used to exclude bacteria and soft tissues from bone graft sites in oral/maxillofacial bone graft sites by using a barrier membrane to maintain the graft contour and space. Current clinical barrier membrane materials based on expanded polytetrafluoroethylene (ePTFE) and bovine type 1 collagen are non-ideal and experience a number of disadvantages including membrane exposure, bacterial colonization/biofilm formation and premature degradation, all of which result in increased surgical intervention and poor bone regeneration. These materials do not actively participate in tissue regeneration, however bioactive materials, such as chitosan, may provide advantages such as the ability to stimulate wound healing and de novo bone formation. Our hypothesis is that electrospun chitosan GTR membranes will support cell attachment and growth but prevent cell infiltration/penetration of membrane, demonstrate in vitro degradation predictive of 4--6 month in vivo functionality, and will deliver antibiotics locally to prevent/inhibit periopathogenic complications. To test this hypothesis a series of chitosan membranes were electrospun, in the presence or absence of genipin, a natural crosslinking agent, at concentrations of 5 and 10 mM. These membranes were characterized by scanning electron microscopy, tensile testing, suture pullout testing, Fourier transform infrared spectroscopy, X-ray diffraction, and gel permeation chromatography, and in vitro biodegradation for diameter/morphology of fibers, membrane strengths, degree of crosslinking, crystallinity, molecular weight, and degradation kinetics, respectively. Cytocompability of membranes was evaluated in osteoblastic, fibroblastic and monocyte cultures. The activity of minocycline loaded and released from the membranes was determined in zone of inhibition tests using P. gingivalis microbe. The results demonstrated that genipin crosslinking extended the in vitro

  14. In vitro evaluation of various bioabsorbable and nonresorbable barrier membranes for guided tissue regeneration

    Directory of Open Access Journals (Sweden)

    Smeets Ralf

    2008-10-01

    Full Text Available Abstract Background Different types of bioabsorbable and nonresorbable membranes have been widely used for guided tissue regeneration (GTR with its ultimate goal of regenerating lost periodontal structures. The purpose of the present study was to evaluate the biological effects of various bioabsorbable and nonresorbable membranes in cultures of primary human gingival fibroblasts (HGF, periodontal ligament fibroblasts (PDLF and human osteoblast-like (HOB cells in vitro. Methods Three commercially available collagen membranes [TutoDent® (TD, Resodont® (RD and BioGide® (BG] as well as three nonresorbable polytetrafluoroethylene (PTFE membranes [ACE (AC, Cytoplast® (CT and TefGen-FD® (TG] were tested. Cells plated on culture dishes (CD served as positive controls. The effect of the barrier membranes on HGF, PDLF as well as HOB cells was assessed by the Alamar Blue fluorometric proliferation assay after 1, 2.5, 4, 24 and 48 h time periods. The structural and morphological properties of the membranes were evaluated by scanning electron microscopy (SEM. Results The results showed that of the six barriers tested, TD and RD demonstrated the highest rate of HGF proliferation at both earlier (1 h and later (48 h time periods (P P ≤ 0.001. In HOB cell culture, the highest rate of cell proliferation was also calculated for TD at all time periods (P Conclusion Results from the present study suggested that GTR membrane materials, per se, may influence cell proliferation in the process of periodontal tissue/bone regeneration. Among the six membranes examined, the bioabsorbable membranes demonstrated to be more suitable to stimulate cellular proliferation compared to nonresorbable PTFE membranes.

  15. Preparation and Characterization of Soluble Eggshell Membrane Protein/PLGA Electro spun Nano fibers for Guided Tissue Regeneration Membrane

    International Nuclear Information System (INIS)

    Jia, J.; Liu, G.; Duan, Y.; Guo, Z.; Yu, J.

    2012-01-01

    Guided tissue regeneration (GTR) is a widely used method in periodontal therapy, which involves the placement of a barrier membrane to exclude migration of epithelium and ensure repopulation of periodontal ligament cells. The objective of this study is to prepare and evaluate a new type of soluble eggshell membrane protein (SEP)/poly (lactic-co-glycolic acid) (PLGA) nano fibers using electro spinning method for GTR membrane application. SEP/PLGA nano fibers were successfully prepared with various blending ratios. The morphology, chemical composition, surface wettability, and mechanical properties of the nano fibers were characterized using scanning electron microscopy (SEM), contact angle measurement, Fourier transform-infrared spectroscopy (FTIR), and a universal testing machine. L-929 fibroblast cells were used to evaluate the biocompatibility of SEP/PLGA nano fibers and investigate the interaction between cells and nano fibers. Results showed that the SEP/PLGA electro spun membrane was composed of uniform, bead-free nano fibers, which formed an interconnected porous network structure. Mechanical property of SEP has been greatly improved by the addition of PLGA. The biological study results showed that SEP/PLGA nano fibers could enhance cell attachment, spreading, and proliferation. The study indicated the potential of SEP/PLGA nano fibers for GTR application and provided a basis for future optimization

  16. Improving the mechanical properties of collagen-based membranes using silk fibroin for corneal tissue engineering.

    Science.gov (United States)

    Long, Kai; Liu, Yang; Li, Weichang; Wang, Lin; Liu, Sa; Wang, Yingjun; Wang, Zhichong; Ren, Li

    2015-03-01

    Although collagen with outstanding biocompatibility has promising application in corneal tissue engineering, the mechanical properties of collagen-based scaffolds, especially suture retention strength, must be further improved to satisfy the requirements of clinical applications. This article describes a toughness reinforced collagen-based membrane using silk fibroin. The collagen-silk fibroin membranes based on collagen [silk fibroin (w/w) ratios of 100:5, 100:10, and 100:20] were prepared by using silk fibroin and cross-linking by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. These membranes were analyzed by scanning electron microscopy and their optical property, and NaCl and tryptophan diffusivity had been tested. The water content was found to be dependent on the content of silk fibroin, and CS10 membrane (loading 10 wt % of silk fibroin) performed the optimal mechanical properties. Also the suture experiments have proved CS10 has high suture retention strength, which can be sutured in rabbit eyes integrally. Moreover, the composite membrane proved good biocompatibility for the proliferation of human corneal epithelial cells in vitro. Lamellar keratoplasty shows that CS10 membrane promoted complete epithelialization in 35 ± 5 days, and their transparency is restored quickly in the first month. Corneal rejection reaction, neovascularization, and keratoconus are not observed. The composite films show potential for use in the field of corneal tissue engineering. © 2014 Wiley Periodicals, Inc.

  17. Expanded polytetrafluoroethylene membrane alters tissue response to implanted Ahmed glaucoma valve.

    Science.gov (United States)

    DeCroos, Francis Char; Ahmad, Sameer; Kondo, Yuji; Chow, Jessica; Mordes, Daniel; Lee, Maria Regina; Asrani, Sanjay; Allingham, R Rand; Olbrich, Kevin C; Klitzman, Bruce

    2009-07-01

    Long-term intraocular pressure control by glaucoma drainage implants is compromised by the formation of an avascular fibrous capsule that surrounds the glaucoma implant and increases aqueous outflow resistance. It is possible to alter this fibrotic tissue reaction and produce a more vascularized and potentially more permeable capsule around implanted devices by enclosing them in a porous membrane. Ahmed glaucoma implants modified with an outer 5-microm pore size membrane (termed porous retrofitted implant with modified enclosure or PRIME-Ahmed) and unmodified glaucoma implants were implanted into paired rabbit eyes. After 6 weeks, the devices were explanted and subject to histological analysis. A tissue response containing minimal vascularization, negligible immune response, and a thick fibrous capsule surrounded the unmodified Ahmed glaucoma implant. In comparison, the tissue response around the PRIME-Ahmed demonstrated a thinner fibrous capsule (46.4 +/- 10.8 microm for PRIME-Ahmed versus 94.9 +/- 21.2 microm for control, p vascularized near the tissue-material interface. A prominent chronic inflammatory response was noted as well. Encapsulating the aqueous outflow pathway with a porous membrane produces a more vascular tissue response and thinner fibrous capsule compared with a standard glaucoma implant plate. Enhanced vascularity and a thinner fibrous capsule may reduce aqueous outflow resistance and improve long-term glaucoma implant performance.

  18. In Vitro Cardiomyogenic Potential of Human Amniotic Fluid Stem Cells

    OpenAIRE

    Guan, Xuan; Delo, Dawn M.; Atala, Anthony; Soker, Shay

    2011-01-01

    Stem cell therapy for damaged cardiac tissue is currently limited by a number of factors, including the inability to obtain sufficient cell numbers, the potential tumorigenicity of certain types of stem cells, and the possible link between stem cell therapy and the development of malignant arrhythmias. In this study, we investigated whether human amniotic fluid-derived stem (hAFS) cells could be a potential source of cells for cardiac cell therapy by testing the in vitro differentiation capab...

  19. In vitro aging of mineralized collagen-based composite as guided tissue regeneration membrane

    Energy Technology Data Exchange (ETDEWEB)

    Pan, S.X. [Department of Prothodontics, School of Stomatology, Peking University, Beijing 100875 (China)]. E-mail: sx_pan@sina.com; Li, Y. [Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China); Feng, H.L. [Department of Prothodontics, School of Stomatology, Peking University, Beijing 100875 (China); Bai, W. [Department of Prothodontics, School of Stomatology, Peking University, Beijing 100875 (China); Gu, Y.Y. [Department of Materials Science and Engineering, Tsinghua University, Beijing 100084 (China)

    2006-05-15

    The technique of guided tissue regeneration (GTR) has been developed for the regeneration of periodontal tissues, bone around natural teeth and dental implants. The aim of this study is to investigate the biodegradability and mechanic behavior of a novel mineralized nano-hydroxyapatite/collagen/poly (lactic acid) (nHAC/PLA) composite as GTR membrane in vitro. The elastic modulus and maximum tensile strength of GTR film samples with different nHAC/PLA ratio were measured to get an optimal nHAC/PLA ratio. Thermogravimetric analysis was conducted to evaluate the change of the inorganic component in the samples during the process of in vitro aging. Morphology of samples was checked by using scanning electron microscopy. On the basis of the above results, it can be concluded that the GTR membranes maintained integrity and the original appearance throughout the 1-month in vitro aging. There is an active dissolution and deposition process of crystals which is propitious to the bone formation on the surface of the composite membrane. The optimal nHAC/PLA ratio of the novel membrane is 0.4:1. For a longer period of bone repair, PLA with higher molecular weight should be chosen as the scaffold for the GTR membrane.

  20. Membrane-reinforced three-dimensional electrospun silk fibroin scaffolds for bone tissue engineering

    International Nuclear Information System (INIS)

    Yang, Sung Yeun; Hwang, Tae Heon; Ryu, WonHyoung; Che, Lihua; Oh, Jin Soo; Ha, Yoon

    2015-01-01

    Electrospun silk fibroin (SF) scaffolds have drawn much attention because of their resemblance to natural tissue architecture such as extracellular matrix, and the biocompatibility of SF as a candidate material to replace collagen. However, electrospun scaffolds lack the physical integrity of bone tissue scaffolds, which require resistance to mechanical loadings. In this work, we propose membrane-reinforced electrospun SF scaffolds by a serial process of electrospinning and freeze-drying of SF solutions in two different solvents: formic acid and water, respectively. After wet electrospinning followed by replacement of methanol with water, SF nanofibers dispersed in water were mixed with aqueous SF solution. Freeze-drying of the mixed solution resulted in 3D membrane-connected SF nanofibrous scaffolds (SF scaffolds) with a thickness of a few centimeters. We demonstrated that the SF concentration of aqueous SF solution controlled the degree of membrane reinforcement between nanofibers. It was also shown that both increase in degree of membrane reinforcement and inclusion of hydroxyapatite (HAP) nanoparticles resulted in higher resistance to compressive loadings of the SF scaffolds. Culture of human osteoblasts on collagen, SF, and SF-HAP scaffolds showed that both SF and SF-HAP scaffolds had biocompatibility and cell proliferation superior to that of the collagen scaffolds. SF-HAP scaffolds with and without BMP-2 were used for in vivo studies for 4 and 8 weeks, and they showed enhanced bone tissue formation in rat calvarial defect models. (paper)

  1. Proteomic profiling of the amniotic fluid to detect inflammation, infection, and neonatal sepsis.

    Directory of Open Access Journals (Sweden)

    Catalin S Buhimschi

    2007-01-01

    Full Text Available Proteomic analysis of amniotic fluid shows the presence of biomarkers characteristic of intrauterine inflammation. We sought to validate prospectively the clinical utility of one such proteomic profile, the Mass Restricted (MR score.We enrolled 169 consecutive women with singleton pregnancies admitted with preterm labor or preterm premature rupture of membranes. All women had a clinically indicated amniocentesis to rule out intra-amniotic infection. A proteomic fingerprint (MR score was generated from fresh samples of amniotic fluid using surface-enhanced laser desorption ionization (SELDI mass spectrometry. Presence or absence of the biomarkers of the MR score was interpreted in relationship to the amniocentesis-to-delivery interval, placental inflammation, and early-onset neonatal sepsis for all neonates admitted to the Newborn Special Care Unit (n = 104. Women with "severe" amniotic fluid inflammation (MR score of 3 or 4 had shorter amniocentesis-to-delivery intervals than women with "no" (MR score of 0 inflammation or even "minimal" (MR score of 1 or 2 inflammation (median [range] MR 3-4: 0.4 d [0.0-49.6 d] versus MR 1-2: 3.8 d [0.0-151.2 d] versus MR 0: 17.0 d [0.1-94.3 d], p 100 cells/mm3, whereas the combination of Gram stain and MR score was best for rapid prediction of intra-amniotic infection (positive amniotic fluid culture.High MR scores are associated with preterm delivery, histological chorioamnionitis, and early-onset neonatal sepsis. In this study, proteomic analysis of amniotic fluid was shown to be the most accurate test for diagnosis of intra-amniotic inflammation, whereas addition of the MR score to the Gram stain provides the best combination of tests to rapidly predict infection.

  2. Alpha-fetoprotein as a tool to distinguish amniotic fluid from urine, vaginal discharge, and semen.

    Science.gov (United States)

    Mor, Amir; Tal, Reshef; Haberman, Shoshana; McCalla, Sandra; Irani, Mohamad; Perlman, Jaqueline; Seifer, David B; Minkoff, Howard

    2015-02-01

    To estimate whether alpha-fetoprotein (AFP) can be used to distinguish amniotic fluid absorbed in sanitary pads from other similarly absorbed substances (semen, urine, and normal vaginal discharge). A prospective cohort study. Urine and amniotic fluid specimens were collected from 52 pregnant women admitted for labor. Semen specimens were collected from 17 men undergoing infertility evaluation. Alpha-fetoprotein concentrations were measured directly from urine, amniotic fluid, and semen and from pads instilled with samples from these specimens. Alpha-fetoprotein concentrations were also measured from pads absorbed with normal vaginal discharge collected from 27 pregnant women. Alpha-fetoprotein levels in amniotic fluid (245.38 ± 21.03 ng/mL, n = 52) were significantly higher than those measured in maternal urine (0.84 ± 0.17 ng/mL, n = 52, P < .001), or semen (1.52 ± 0.35 ng/mL, n = 17, P < .001). The same trend was seen when AFP was extracted from pads: amniotic fluid levels (19.44 ± 1.98 ng/mL, n=52) were significantly higher than those of urine (undetectable, n=52), semen (undetectable, n = 17), or normal vaginal discharge (0.53 ± 0.16 ng/mL, n = 27, P < .001). Receiver operator characteristic curve analysis demonstrated 96.2% sensitivity and 100% specificity for distinguishing the presence of amniotic fluid from normal vaginal discharge on sanitary pads (cutoff 3.88 ng/mL, area under the curve 0.99). When the diagnosis of rupture of membranes is in doubt, AFP levels can assist in differentiating amniotic fluid from other bodily fluids. A method that utilizes sanitary pads and an assay for AFP quantification may be an accurate and convenient way to confirm the diagnosis of rupture of membranes.

  3. Importance of banked tissues in the management of mass nuclear casualties

    International Nuclear Information System (INIS)

    Singh, Rita; Bhatnagar, P.K.

    2009-01-01

    Nuclear detonations are the most devastating of the weapons of mass destruction. There will be large number of casualties on detonation of nuclear weapon. Biological tissues like bone, skin, amniotic membrane and other soft tissues can be used for repair or reconstruction of the injured part of the body. Tissues from human donor can be processed and banked for orthopaedic, spinal, trauma and other surgical procedures. Radiation technology is used to sterilize the tissues to make them safe for clinical use. This paper highlights the importance of such banked tissues in the management of the casualties. (author)

  4. Sterilization of biological tissues with ionizing radiation

    International Nuclear Information System (INIS)

    Reyes F, M.L.; Martinez P, M.E.; Luna Z, D.

    1997-01-01

    On June 1994, the National Institute of Nuclear Research (ININ) and the South Central Hospital for High Specialty of PEMEX (HCSAE) began a joint work with the finality to obtain radio sterilized amniotic membranes for to be used as cover (biological bandage) in burnt patients. Subsequently the Chemistry Faculty of UNAM and the National Institute of Cardiology began to collaborate this last with interest on cardiac valves for graft. Starting from 1997, the International Atomic Energy Agency (IAEA) supports this project (MEX/7/008) whose main objective is to set up the basis to establish in Mexico a Radio sterilized Tissue Bank (amniotic membranes, skin, bones, tendons, cardiac valves, etc.) to be used with therapeutic purposes (grafts). The IAEA support has consisted in the equipment acquisition which is fundamental for the Tissue Bank performance such as an experimental irradiator, laminar flow bell, lyophilizer, vacuum sealer and special knives for tissues. Also visits to Mexico of experts have been authorized with the aim of advising to the personnel which participate in the project and scientific visits of this personnel to another tissue banks (Sri Lanka and Argentine). The establishment in Mexico of a Tissue bank will be a great benefit because it will have availability of distinct tissues for grafts and it will reduce the synthetic materials importation which is very expensive. (Author)

  5. [Application of the xenogenic acellular dermal matrix membrane application used in the postoperative tissue shortage repair].

    Science.gov (United States)

    Bai, Yanxia; Yan, Liying; Zhang, Shaoqiang; Shao, Yuan; Yao, Xiaobao; Li, Honghui; Zhao, Ruimin; Zhao, Qian; Zhang, Pengfei; Yang, Qi

    2014-09-01

    To observe the short-term and long-term curative effect of the xenogenic acellular dermal matrix membrane (or joint muscle flap transfer) application used in the 82 cases postoperative tissue shortage repair that after the head neck carcinoma resection. To held the 82 cases head neck carcinoma postoperative mucosa shortage repaired after resection by the xenogenic acellular dermal matrix membrane (or joint muscle flap transfer), 65 cases mucosa shortage wound be directly covered by the repair membrane and the other 17 cases mucosa shortage wound be repaired by the tranfered muscle tissue flap with the repair membrane covered; 53 cases underwent additional postoperative radiotherapy between 2-4 weeks and follow-up in 1, 3, 6, 12, 18, 24, 30, 36, 48, 60 months and observed the operation site repair process through the electronic laryngoscope, observed the patients respiration, swallow, phonation function. Seventy-seven cases patients operation incision reached I phase healing standard, another 5 cases patients operation incision reached II phase healing standard because of the wound infection and fully-recovered through the local wound drainage,dressing process. All the patients tracheal cannula,the stomach tube be extubated successfully and without the local cicatricial constriction occurred. Seventy-eight cases follow up period reached 1 year including 53 cases who underwent postoperative radiotherapy, 49 cases follow up period reached 3 years including 32 cases who underwent postoperative radiotherapy, 14 cases follow up period reached 5 years including 12 cases who underwent postoperative radiotherapy. The patients with static local lesions discovered no reaction such as exclusion, allergy. The application of xenogenic acellular dermal matrix membrane (or joint muscle flap transfer used in in the postoperative tissue shortage repair that after the head neck carcinoma resection have several advantage such as comparatively easily implementation, operation safety

  6. Preparation of biodegradable PLA/PLGA membranes with PGA mesh and their application for periodontal guided tissue regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Eun Jin; Kang, Inn-Kyu [Department of Polymer Science, Kyungpook National University, 1370 Sankyuk-dong, Buk-gu, Daegu 702-701 (Korea, Republic of); Yoon, Suk Joon [Department of Biology, Sookmyung Women' s University, Hyochangwongil 52, Yongsan-gu, Seoul 140-742 (Korea, Republic of); Yeo, Guw-Dong; Pai, Chaul-Min, E-mail: ikkang@knu.ac.k [Samyang Central R and D Center, 63-2 Hwaam-dong, Yusung-gu, Daejeon 305-717 (Korea, Republic of)

    2009-10-15

    A biodegradable polylactic acid (PLA)/poly(glycolide-co-lactide) copolymer (PLGA) membrane with polyglycolic acid (PGA) mesh was prepared to aid the effective regeneration of defective periodontal tissues. The microporous membrane used in this study consists of biodegradable polymers, and seems to have a structure to provide appropriate properties for periodontal tissue regeneration. Based on the albumin permeation test, it is known that the biodegradable membrane exhibits the suitable permeability of nutrients. The membrane maintained its physical integrity for 6-8 weeks, which could be sufficient to retain space in the periodontal pocket. Cell attachment and cytotoxicity tests were performed with respect to the evaluation of biocompatibility of the membrane. As a result, the membrane did not show any cytotoxicity. The safety and therapeutic efficacies of the biodegradable membranes were confirmed in animal tests.

  7. Amniotic Band Syndrome, Perinatal Hospice, and Palliative Care versus Active Management

    Directory of Open Access Journals (Sweden)

    Shadi Rezai

    2016-01-01

    Full Text Available Introduction. Amniotic band syndrome and sequence are a relatively rare condition in which congenital anomalies occur as a result of the adherence and entrapment of fetal parts with coarse fibrous bands of the amniotic membrane. A large percentage of reported cases have an atypical gestational history. The frequency of this obstetric complication is not affected by fetal gender, genetic abnormality, or prenatal infection. Case. A 21-year-old, G1P0 female parturient at 18 weeks and 5 days with a single intrauterine gestation during a routine ultrasound evaluation was noted to have amniotic band sequence. The pregnancy was subsequently complicated by preterm premature rupture of membranes with oligohydramnios, resulting in a surviving neonate scheduled for rehabilitative treatment. Conclusion. Amniotic band syndrome is an uncommon congenital anomaly resulting in multiple disfiguring and disabling manifestations. Several theories are proposed with most involving early rupture of the amnion and entanglement of fetal parts by amniotic bands. This syndrome can be manifested by development of multiple malformations, with the majority of the defects being limb abnormalities of a disorganized nature, as in the case we present. In the absence of a clear etiology of consequential congenital abnormalities, obstetric management guidelines should use shared decision models to focus on the quality of life for the offspring.

  8. Synthesis of chitosan/hydroxyapatite membranes coated with hydroxycarbonate apatite for guided tissue regeneration purposes

    Energy Technology Data Exchange (ETDEWEB)

    Fraga, Alexandre Felix [Federal University of Sao Carlos, Department of Materials Engineering, DEMa, UFSCar, Sao Carlos, SP (Brazil); Almeida Filho, Edson de, E-mail: edsonafilho@yahoo.com.br [University Estadual Paulista, Department of Physical Chemistry - IQ, Araraquara, SP (Brazil); Silva Rigo, Eliana Cristina da [University of Sao Paulo, Department of Basic Science - FZEA-ZAB, Pirassununga, SP (Brazil); Ortega Boschi, Anselmo [Federal University of Sao Carlos, Department of Materials Engineering, DEMa, UFSCar, Sao Carlos, SP (Brazil)

    2011-02-15

    Chitosan, which is a non-toxic, biodegradable and biocompatible biopolymer, has been widely researched for several applications in the field of biomaterials. Calcium phosphate ceramics stand out among the so-called bioceramics for their absence of local or systemic toxicity, their non-response to foreign bodies or inflammations, and their apparent ability to bond to the host tissue. Hydroxyapatite (HA) is one of the most important bioceramics because it is the main component of the mineral phase of bone. The aim of this work was to produce chitosan membranes coated with hydroxyapatite using the modified biomimetic method. Membranes were synthesized from a solution containing 2% of chitosan in acetic acid (weight/volume) via the solvent evaporation method. Specimens were immersed in a sodium silicate solution and then in a 1.5 SBF (simulated body fluid) solution. The crystallinity of the HA formed over the membranes was correlated to the use of the nucleation agent (the sodium silicate solution itself). Coated membranes were characterized by means of scanning electron microscopy - SEM, X-ray diffraction - XRD, and Fourier transform infrared spectroscopy - FTIR. The results indicate a homogeneous coating covering the entire surface of the membrane and the production of a semi-crystalline hydroxyapatite layer similar to the mineral phase of human bone.

  9. Biochemical characterization of the plasma membrane H+ - ATPase from red beet (Beta vulgaris) hypocotyl tissue

    International Nuclear Information System (INIS)

    Oleski, N.A.

    1986-01-01

    Several biochemical techniques including selective solubilization followed by gel filtration or various types of affinity chromatography, and antibody production were employed in an attempt to purify the plasma membrane H + - ATPase from red beet hypocotyl tissue. While the enzyme could not be purified using any of these methods, it was possible to successfully conduct a more detailed biochemical analysis of the H + - ATPase. The molecular weight and isoelectric point of the enzyme were determined using N,N'dicyclohexylcarbodiimide (DCCD) and a H + - ATPase antibody. When plasma membrane vesicles were incubated with 20 μM [ 14 C]-DCCD at 0 C, a single 97,000 dalton protein was apparent on a fluorograph. A close correlation between [ 14 C]-DCCD labelling of the 97,000 dalton protein and the extent of ATPase inhibition over a range of DCCD concentrations suggests that this 97,000 dalton protein is a component of the plasma membrane H + - ATPase. An antibody raised against the plasma membrane H + - ATPase of Neurospora crassa cross-reacted with the 97,000 dalton DCCD-binding protein, further supporting the identity of this protein. Immunoblots of two dimensional gels of red beet plasma membrane vesicles indicated the isoelectric point of the enzyme to be pH 6.5

  10. Prenatal diagnosis of amniotic band syndrome

    Directory of Open Access Journals (Sweden)

    Laxmi Devi Padmanabhan

    2016-01-01

    Full Text Available Amniotic band can cause a broad spectrum of anomalies ranging from simple band constrictions to major craniofacial and visceral defects. It can cause significant neonatal morbidity. Accurate diagnosis will help in the management of the present pregnancy and in counseling with regard to future pregnancies. Here we report three cases of amniotic band syndrome detected in the prenatal period.

  11. Tissue Factor Coagulant Activity is Regulated by the Plasma Membrane Microenvironment.

    Science.gov (United States)

    Yu, Yuanjie; Böing, Anita N; Hau, Chi M; Hajji, Najat; Ruf, Wolfram; Sturk, Auguste; Nieuwland, Rienk

    2018-06-01

    Tissue factor (TF) can be present in a non-coagulant and coagulant form. Whether the coagulant activity is affected by the plasma membrane microenvironment is unexplored.  This article studies the presence and coagulant activity of human TF in plasma membrane micro-domains.  Plasma membranes were isolated from human MIA PaCa2 cells, MDA-MB-231 cells and human vascular smooth muscle cells by Percoll gradient ultracentrifugation after cell disruption. Plasma membranes were fractionated by OptiPrep gradient ultracentrifugation, and the presence of TF, flotillin, caveolin, clathrin, protein disulphide isomerase (PDI), TF pathway inhibitor (TFPI) and phosphatidylserine (PS) were determined.  Plasma membranes contain two detergent-resistant membrane (DRM) compartments differing in density and biochemical composition. High-density DRMs (DRM-H) have a density ( ρ ) of 1.15 to 1.20 g/mL and contain clathrin, whereas low-density DRMs (DRM-L) have a density between 1.09 and 1.13 g/mL and do not contain clathrin. Both DRMs contain TF, flotillin and caveolin. PDI is detectable in DRM-H, TFPI is not detectable in either DMR-H or DRM-L and PS is detectable in DRM-L. The DRM-H-associated TF (> 95% of the TF antigen) lacks detectable coagulant activity, whereas the DRM-L-associated TF triggers coagulation. This coagulant activity is inhibited by lactadherin and thus PS-dependent, but seemed insensitive to 16F16, an inhibitor of PDI.  Non-coagulant and coagulant TF are present within different types of DRMs in the plasma membrane, and the composition of these DRMs may affect the TF coagulant activity. Schattauer GmbH Stuttgart.

  12. Drug loaded homogeneous electrospun PCL/gelatin hybrid nanofiber structures for anti-infective tissue regeneration membranes.

    Science.gov (United States)

    Xue, Jiajia; He, Min; Liu, Hao; Niu, Yuzhao; Crawford, Aileen; Coates, Phil D; Chen, Dafu; Shi, Rui; Zhang, Liqun

    2014-11-01

    Infection is the major reason for guided tissue regeneration/guided bone regeneration (GTR/GBR) membrane failure in clinical application. In this work, we developed GTR/GBR membranes with localized drug delivery function to prevent infection by electrospinning of poly(ε-caprolactone) (PCL) and gelatin blended with metronidazole (MNA). Acetic acid (HAc) was introduced to improve the miscibility of PCL and gelatin to fabricate homogeneous hybrid nanofiber membranes. The effects of the addition of HAc and the MNA content (0, 1, 5, 10, 20, 30, and 40 wt.% of polymer) on the properties of the membranes were investigated. The membranes showed good mechanical properties, appropriate biodegradation rate and barrier function. The controlled and sustained release of MNA from the membranes significantly prevented the colonization of anaerobic bacteria. Cells could adhere to and proliferate on the membranes without cytotoxicity until the MNA content reached 30%. Subcutaneous implantation in rabbits for 8 months demonstrated that MNA-loaded membranes evoked a less severe inflammatory response depending on the dose of MNA than bare membranes. The biodegradation time of the membranes was appropriate for tissue regeneration. These results indicated the potential for using MNA-loaded PCL/gelatin electrospun membranes as anti-infective GTR/GBR membranes to optimize clinical application of GTR/GBR strategies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Tissue eosinophilia induced by recombinant human interleukin-5 in the hamster cheek pouch membrane

    Directory of Open Access Journals (Sweden)

    M. Minnicozzi

    1995-01-01

    Full Text Available Interleukin-5 (IL-5 is a cytokine that preferentially effects the development and function of eosinophils, and is considered important in the pathophysiology of allergic inflammation. In this study, we evaluated the ability of recombinant human IL-5 (rHu IL-5 to promote tissue eosinophilia and the importance of this eosinophilia to pathological alterations in vascular function. Repetitive subcutaneous administration for 18 days of rHu IL-5 resulted in a 7-fold increase in the number of eosinophils found in the ipsilateral hamster cheek pouch membrane. The contralateral cheek pouch membrane and peritoneum of these animals showed lesser but significant elevations in the number of eosinophils. In contrast, denatured rHu IL-5 did not elevate eosinophils in these tissues. Through the use of intravital microscopy and fluorometric analysis, rHu IL-5 treated hamster cheek pouch membranes were evaluated for alterations in microvascular permeability, using plasma clearance of FITC-dextran 150 as an index. Despite promoting a prominent tissue eosinophilia, the repetitive subcutaneous injections of rHu IL-5 did not alter the clearance of FITC-dextran 150. Topical application of rHu IL-5 to the cheek pouch, also, had no effect on the clearance of FITC-dextran 150. Immunofluorescence observations using an antibody to the granule protein, eosinophil peroxidase, indicated that the recruited cells had not degranulated. Our results support the importance of IL-5 in the recruitment of tissue eosinophils, but further stimulation is probably required to cause degranulation of these cells and the ensuing tissue damage.

  14. Surgical tissue banking - the role of nuclear technology

    International Nuclear Information System (INIS)

    Hasim Mohamad

    2002-01-01

    Before 1980s amniotic membranes were mainly used clinically either in fresh forms or preserved in glycerol without sterilization. There were problems of graft infections and difficulty in maintaining standard of graft quality and uniformity of the amnion when applied on the recipient patients. Nuclear technology has helped to sterilize these membranes such that they can be processed, dried and then double packed before finally irradiated with gamma rays emitted from the radionuclide cobalt-60. The irradiated amniotic membranes can be kept at room temperature and are readily available for used by doctors without relying on glycerol for preservation or freezers for storage. Similarly allograft bones can be processed and be stored in similar manner otherwise they need to be frozen at -80 to -160 degree C. Radiation technology has benefited many patients in many countries in the Asia Pacific region through sterilization of tissues. Bone such as femoral heads can be processed into radiation sterilized bone chips or grounded bones and has been successfully used for filling up of cavities left by benign bone tumors, bone cysts etc. These materials are safe and cheap and save the recipients in-term of money and a second surgery and on the extreme case, patients may avoid amputation if an irradiated long bone is available. Orthopedic, neuro- and dental surgeons have reported many such successful outcomes. Also bum patients have been shown to benefit from amniotic membranes or porcine skin which were processed and irradiated prior to treatment. In some countries in this region, amniotic membranes have been used as en effective wound dressing for leprosy patients as well. Reports of these cases are to be presented. (Author)

  15. Psychobiology of the amniotic environment.

    Science.gov (United States)

    Benassi, Luigi; Accorsi, Francesca; Marconi, Lorenza; Benassi, Gianluca

    2004-01-01

    Water, basic element of amniotic fluid (A.F.), is closely related to Life, Fertility and Motherhood in several cultures and religions. Through material evidences of an essential growth medium and useful diagnostic source, a new concept grow up: the fluid as a first real environment in which fetus lives and acts. Many studies confirm that in A.F. fetus starts his character-building, his memory and his intelligence. The fluid seems to be the first means of learning and acknowledgement. Sounds, smells and tastes are perceived as well as emotions and fears. Urinoterapy and staminal cells sampling shows how A.F. can be considered as an additional terapeutic resource.

  16. Artificial membrane-binding proteins stimulate oxygenation of stem cells during engineering of large cartilage tissue

    Science.gov (United States)

    Armstrong, James P. K.; Shakur, Rameen; Horne, Joseph P.; Dickinson, Sally C.; Armstrong, Craig T.; Lau, Katherine; Kadiwala, Juned; Lowe, Robert; Seddon, Annela; Mann, Stephen; Anderson, J. L. Ross; Perriman, Adam W.; Hollander, Anthony P.

    2015-06-01

    Restricted oxygen diffusion can result in central cell necrosis in engineered tissue, a problem that is exacerbated when engineering large tissue constructs for clinical application. Here we show that pre-treating human mesenchymal stem cells (hMSCs) with synthetic membrane-active myoglobin-polymer-surfactant complexes can provide a reservoir of oxygen capable of alleviating necrosis at the centre of hyaline cartilage. This is achieved through the development of a new cell functionalization methodology based on polymer-surfactant conjugation, which allows the delivery of functional proteins to the hMSC membrane. This new approach circumvents the need for cell surface engineering using protein chimerization or genetic transfection, and we demonstrate that the surface-modified hMSCs retain their ability to proliferate and to undergo multilineage differentiation. The functionalization technology is facile, versatile and non-disruptive, and in addition to tissue oxygenation, it should have far-reaching application in a host of tissue engineering and cell-based therapies.

  17. Effects of topical 0.2% Cyclosporine A on corneal neovascularization induced by xenologous amniotic membrane implantation into a corneal stroma micropocket of rats Efeitos do uso tópico da Ciclosporina A (CsA 0,2% na neovascularização corneal induzida pelo implante de membrana amniótica xenógena em microbolsa no estroma da córnea de ratos

    Directory of Open Access Journals (Sweden)

    Juliana F. Milani

    2008-08-01

    Full Text Available The objective of the study was to evaluate the topical effects of 0.2% Cyclosporine A (CsA on corneal neovascularization of rats following surgical implantation of equine amniotic membrane into a corneal stroma micropocket. The implantation of xenologous amniotic membrane was performed bilaterally in 90 rats. In the same day of the surgery each right eye started receiving topical CsA twice a day. The left eye received no medication and served as a control. The evaluation of corneal neovascularization was performed by computerized image analysis and histopathological evaluation at 1, 3, 7, 15, 30 and 60 days postoperatively. For the image analysis 10 animals were used per time period, and for the histopathological examination, five animals were used per time period. Image analysis found that corneal neovascularization began on the 3rd postoperative day, reached its peak on the 7th day, and then progressively and rapidly decreased. Statistic analysis indicated that neovascularization of the CsA treated eye on the 7th day was significantly higher than that observed in untreated eyes. On the 30th day, however, this pattern was reversed with the neovascularization observed in the CsA treated eyes declining to the low levels observed on the 3rd day. The degree of neovascularization in the untreated eyes on the 30th day declined to the baseline levels found on day 3 at the 60th day. Histopathological analysis indicated that deposition of collagen in the implanted tissue was completed by the 15th day. Therefore, we concluded that (1 equine amniotic membrane in rat corneal stroma produced an intense neovascularization until the 15th day postoperatively and then regressed, (2 deposition of collagen of the implanted tissue was completed on the 15th day postoperatively, and (3 use of CsA was associated with increase in the corneal neovascularization initially, followed by a quick and intense regression.Este estudo teve como objetivo a avaliação dos

  18. Amniotic fluid chemokines and autism spectrum disorders: An exploratory study utilizing a Danish Historic Birth Cohort

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Larsen, Nanna Brink; Grove, Jakob

    2012-01-01

    Elevated levels of chemokines have been reported in plasma and brain tissue of individuals with Autism Spectrum Disorders (ASD). The aim of this study was to examine chemokine levels in amniotic fluid (AF) samples of individuals diagnosed with ASD and their controls.......Elevated levels of chemokines have been reported in plasma and brain tissue of individuals with Autism Spectrum Disorders (ASD). The aim of this study was to examine chemokine levels in amniotic fluid (AF) samples of individuals diagnosed with ASD and their controls....

  19. Intra-amniotic pharmacological blockade of inflammatory signalling pathways in an ovine chorioamnionitis model.

    Science.gov (United States)

    Ireland, D J; Kemp, M W; Miura, Y; Saito, M; Newnham, J P; Keelan, J A

    2015-05-01

    Intrauterine inflammation (IUI) associated with infection is the major cause of preterm birth (PTB) at PTBs. Pharmacological strategies to prevent PTB and improve fetal outcomes will likely require both antimicrobial and anti-inflammatory therapies. Here we investigated the effects of two cytokine-suppressive anti-inflammatory drugs (CSAIDs), compounds that specifically target inflammatory signalling pathways, in an ovine model of lipopolysaccharide (LPS)-induced chorioamnionitis. Chronically catheterized ewes at 116 days gestation (n = 7/group) received an intra-amniotic (IA) bolus of LPS (10 mg) plus vehicle or CSAIDS: TPCA-1 (1.2 mg/kg fetal weight) or 5z-7-oxozeaenol (OxZnl; 0.4 mg/kg fetal weight); controls received vehicle (dimethylsulphoxide). Amniotic fluid (AF), fetal and maternal blood samples were taken 0, 2, 6, 12, 24 and 48 h later; tissues were taken at autopsy (48 h). Administration of TPCA-1 or OxZnl abrogated the stimulatory effects of LPS (P < 0.01 versus vehicle control) on production of PGE2 in AF, with lesser (non-significant) effects on IL-6 production. Fetal membrane polymorphonuclear cell infiltration score was significantly higher in LPS versus vehicle control animals (P < 0.01), and this difference was absent with TPCA-1 and OxZnl treatment. LPS-induced systemic fetal inflammation was highly variable, with no significant effects of CSAIDs observed. Lung inflammation was evident with LPS exposure, but unaffected by CSAID treatment. We have shown in a large animal model that IA administration of a single dose of CSAIDs can suppress LPS-induced IA inflammatory responses, while fetal effects were minimal. Further development and investigation of these compounds in infectious models is warranted. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  20. Wet-Lay Process - A Novel Approach to Scalable Fabrication of Tissue Scaffolds and Reinforcement Membranes

    Science.gov (United States)

    Wood, Andrew

    Fibrous materials received a great deal of interest in the fields of tissue engineering and regenerative medicine due to the beneficial cell-interactions and tunable properties for various biomedical applications. These materials are highly advantageous as they provide a large surface area for cellular attachment, proliferation, high porosity values for cellular in-growth, and the ability to modify the membrane to achieve desired responses to both mechanical loading as well as environmental stimuli. A prominent method currently used to fabricate such membranes is electrospinning which uses electrostatic forces to produce fibers on the range of nanometers giving them high morphological saliency to the native extra cellular matrix (ECM). These fibers are also advantageous mechanically with strength and flexibility due to their larger aspect ratio when compared to larger diameter micro/macro fibers. While this spinning technique has many advantages and has seen the most quantity of research in recent years, it does have its own set of drawbacks. Among them is the use cytotoxic solvents during processing which must be fully removed before implantation. In addition, since the fiber produced have smaller diameters, the resulting average pore-size of the scaffold is decreased which in turn hinders cellular penetration into the bulk scaffold. In this work, we have proposed and characterized a novel method called wet-lay process for the rapid fabrication of fibrous membranes for tissue scaffolds. Wet-laying is a method common to textiles and paper industry but unexplored for tissue scaffolds. Short fibers are first suspended in an aqueous bath and homogeneously dispersed using shear force. After draining away the aqueous solution, a nonwoven fibro-porous membrane is deposited onto the draining screen. The implementation of wet-laid membranes into weak hydrogel matrices has shown a reinforcement effect for the composite. Further analyses were carried out to determine the

  1. Cell-Adhesive Bioinspired and Catechol-Based Multilayer Freestanding Membranes for Bone Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Maria P. Sousa

    2017-10-01

    Full Text Available Mussels are marine organisms that have been mimicked due to their exceptional adhesive properties to all kind of surfaces, including rocks, under wet conditions. The proteins present on the mussel’s foot contain 3,4-dihydroxy-l-alanine (DOPA, an amino acid from the catechol family that has been reported by their adhesive character. Therefore, we synthesized a mussel-inspired conjugated polymer, modifying the backbone of hyaluronic acid with dopamine by carbodiimide chemistry. Ultraviolet–visible (UV–Vis spectroscopy and nuclear magnetic resonance (NMR techniques confirmed the success of this modification. Different techniques have been reported to produce two-dimensional (2D or three-dimensional (3D systems capable to support cells and tissue regeneration; among others, multilayer systems allow the construction of hierarchical structures from nano- to macroscales. In this study, the layer-by-layer (LbL technique was used to produce freestanding multilayer membranes made uniquely of chitosan and dopamine-modified hyaluronic acid (HA-DN. The electrostatic interactions were found to be the main forces involved in the film construction. The surface morphology, chemistry, and mechanical properties of the freestanding membranes were characterized, confirming the enhancement of the adhesive properties in the presence of HA-DN. The MC3T3-E1 cell line was cultured on the surface of the membranes, demonstrating the potential of these freestanding multilayer systems to be used for bone tissue engineering.

  2. Multilineage Potential Research of Bovine Amniotic Fluid Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Yuhua Gao

    2014-02-01

    Full Text Available The use of amnion and amniotic fluid (AF are abundant sources of mesenchymal stem cells (MSCs that can be harvested at low cost and do not pose ethical conflicts. In human and veterinary research, stem cells derived from these tissues are promising candidates for disease treatment, specifically for their plasticity, their reduced immunogenicity, and high anti-inflammatory potential. This work aimed to obtain and characterize bovine amniotic fluid mesenchymal stem cells (AFMSC. The bovine AF from the amniotic cavity of pregnant gilts in the early stages of gestation (3- and 4-m-old bovine embryos was collected. AFMSCs exhibit a fibroblastic-like morphology only starting from the fourth passage, being heterogeneous during the primary culture. Immunofluorescence results showed that AFMSCs were positive for β-integrin, CD44, CD73 and CD166, but negative for CD34, CD45. Meanwhile, AFMSCs expressed ES cell markers, such as Oct4, and when appropriately induced, are capable of differentiating into ectodermal and mesodermal lineages. This study reinforces the emerging importance of these cells as ideal tools in veterinary medicine; future studies aimed at a deeper evaluation of their immunological properties will allow a better understanding of their role in cellular therapy.

  3. In Situ Blotting : A Novel Method for Direct Transfer of Native Proteins from Sectioned Tissue to Blotting Membrane

    NARCIS (Netherlands)

    Okabe, Masashi; Nyakas, Csaba; Buwalda, Bauke; Luiten, Paul G.M.

    1993-01-01

    We describe a novel technique for direct transfer of native proteins from unfixed frozen tissue sections to an immobilizing matrix, e.g., nitrocellulose, polyvinyliden difluoride, or positively charged nylon membranes. Proteins are directly blotted onto the membrane, providing optimal accessibility

  4. Asymmetric PDLLA membranes containing Bioglass(R) for guided tissue regeneration: characterization and in vitro biological behavior

    NARCIS (Netherlands)

    Leal, A.I.; Caridade, S.G.; Ma, J.; Yu, N.; Gomes, M.; Reis, R.L.; Jansen, J.A.; Walboomers, X.F.; Mano, J.F.

    2013-01-01

    OBJECTIVE: In the treatment of periodontal defects, composite membranes might be applied to protect the injured area and simultaneously stimulate tissue regeneration. This work describes the development and characterization of poly(d,l-lactic acid)/Bioglass(R) (PDLLA/BG) composite membranes with

  5. Fabrication and characterization of two-layered nanofibrous membrane for guided bone and tissue regeneration application.

    Science.gov (United States)

    Masoudi Rad, Maryam; Nouri Khorasani, Saied; Ghasemi-Mobarakeh, Laleh; Prabhakaran, Molamma P; Foroughi, Mohammad Reza; Kharaziha, Mahshid; Saadatkish, Niloufar; Ramakrishna, Seeram

    2017-11-01

    Membranes used in dentistry act as a barrier to prevent invasion of intruder cells to defected area and obtains spaces that are to be subsequently filled with new bone and provide required bone volume for implant therapy when there is insufficient volume of healthy bone at implant site. In this study a two-layered bioactive membrane were fabricated by electrospinning whereas one layer provides guided bone regeneration (GBR) and fabricated using poly glycerol sebacate (PGS)/polycaprolactone (PCL) and Beta tri-calcium phosphate (β-TCP) (5, 10 and 15%) and another one containing PCL/PGS and chitosan acts as guided tissue regeneration (GTR). The morphology, chemical, physical and mechanical characterizations of the membranes were studied using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), tensile testing, then biodegradability and bioactivity properties were evaluated. In vitro cell culture study was also carried out to investigate proliferation and mineralization of cells on different membranes. Transmission electron microscope (TEM) and SEM results indicated agglomeration of β-TCP nanoparticles in the structure of nanofibers containing 15% β-TCP. Moreover by addition of β-TCP from 5% to 15%, contact angle decreased due to hydrophilicity of nanoparticles and bioactivity was found to increase. Mechanical properties of the membrane increased by incorporation of 5% and 10% of β-TCP in the structure of nanofibers, while addition of 15% of β-TCP was found to deteriorate mechanical properties of nanofibers. Although the presence of 5% and 10% of nanoparticles in the nanofibers increased proliferation of cells on GBR layer, cell proliferation was observed to decrease by addition of 15% β-TCP in the structure of nanofibers which is likely due to agglomeration of nanoparticles in the nanofiber structure. Our overall results revealed PCL/PGS containing 10% β-TCP could be selected as the optimum GBR membrane

  6. Characterization of membranous (M) cells in normal feline conjunctiva-associated lymphoid tissue (CALT).

    Science.gov (United States)

    Giuliano, Elizabeth A; Finn, Kevin

    2011-09-01

    To characterize conjunctival lymphoid nodules obtained from the nictitans of healthy cats to determine if the follicle-associated epithelium (FAE) of conjunctiva-associated lymphoid tissue (CALT) in this species contains membranous (M)-cells analogous to those described in other regions of mucosa-associated lymphoid tissue (MALT). Lymphoid follicles from nictitan bulbar surfaces of 10 healthy cats (20 eyes total) were examined. Nictitans from five cats were harvested immediately post-mortem and a minimum of 12 lymphoid nodules from each third eyelid were isolated using a Zeiss operating microscope. At least three lymphoid follicles from each eye were examined using light microscopy (LM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) using standard fixation and embedding protocols. Nictitan-lymphoid follicles from another five healthy cats were processed for immunohistochemistry to characterize the distribution of T- and B-lymphocytes present beneath the FAE. The FAE overlying CALT from 10 healthy cats demonstrated morphology characteristic of M-cells including attenuated apical cell surface with blunted microvilli and microfolds, invaginated basolateral membrane forming a cytoplasmic pocket, and diminished distance between the apical and pocket membrane. Immunohistochemistry of lymphoid tissue subtending the FAE demonstrated B-cell dependent regions in the germinal centers surrounded by T-cell dependent interfollicular zones. Healthy feline CALT contains morphologic features analogous to those described in other regions of MALT. Documentation of feline conjunctival M-cells is of clinical relevance in the study of primary infectious, allergic, and autoimmune ocular diseases, as well as a potential means of vaccination or drug delivery. © 2011 American College of Veterinary Ophthalmologists.

  7. Multidimensional proteomics analysis of amniotic fluid to provide insight into the mechanisms of idiopathic preterm birth.

    Directory of Open Access Journals (Sweden)

    Irina A Buhimschi

    2008-04-01

    Full Text Available Though recent advancement in proteomics has provided a novel perspective on several distinct pathogenetic mechanisms leading to preterm birth (inflammation, bleeding, the etiology of most preterm births still remains elusive. We conducted a multidimensional proteomic analysis of the amniotic fluid to identify pathways related to preterm birth in the absence of inflammation or bleeding.A proteomic fingerprint was generated from fresh amniotic fluid using surface-enhanced laser desorbtion ionization time of flight (SELDI-TOF mass spectrometry in a total of 286 consecutive samples retrieved from women who presented with signs or symptoms of preterm labor or preterm premature rupture of the membranes. Inflammation and/or bleeding proteomic patterns were detected in 32% (92/286 of the SELDI tracings. In the remaining tracings, a hierarchical algorithm was applied based on descriptors quantifying similarity/dissimilarity among proteomic fingerprints. This allowed identification of a novel profile (Q-profile based on the presence of 5 SELDI peaks in the 10-12.5 kDa mass area. Women displaying the Q-profile (mean+/-SD, gestational age: 25+/-4 weeks, n = 40 were more likely to deliver preterm despite expectant management in the context of intact membranes and normal amniotic fluid clinical results. Utilizing identification-centered proteomics techniques (fluorescence two-dimensional differential gel electrophoresis, robotic tryptic digestion and mass spectrometry coupled with Protein ANalysis THrough Evolutionary Relationships (PANTHER ontological classifications, we determined that in amniotic fluids with Q-profile the differentially expressed proteins are primarily involved in non-inflammatory biological processes such as protein metabolism, signal transduction and transport.Proteomic profiling of amniotic fluid coupled with non-hierarchical bioinformatics algorithms identified a subgroup of patients at risk for preterm birth in the absence of intra-amniotic

  8. Distribution of Basement Membrane Molecules, Laminin and Collagen Type IV, in Normal and Degenerated Cartilage Tissues.

    Science.gov (United States)

    Foldager, Casper Bindzus; Toh, Wei Seong; Gomoll, Andreas H; Olsen, Bjørn Reino; Spector, Myron

    2014-04-01

    The objective of the present study was to investigate the presence and distribution of 2 basement membrane (BM) molecules, laminin and collagen type IV, in healthy and degenerative cartilage tissues. Normal and degenerated tissues were obtained from goats and humans, including articular knee cartilage, the intervertebral disc, and meniscus. Normal tissue was also obtained from patella-tibial enthesis in goats. Immunohistochemical analysis was performed using anti-laminin and anti-collagen type IV antibodies. Human and goat skin were used as positive controls. The percentage of cells displaying the pericellular presence of the protein was graded semiquantitatively. When present, laminin and collagen type IV were exclusively found in the pericellular matrix, and in a discrete layer on the articulating surface of normal articular cartilage. In normal articular (hyaline) cartilage in the human and goat, the proteins were found co-localized pericellularly. In contrast, in human osteoarthritic articular cartilage, collagen type IV but not laminin was found in the pericellular region. Nonpathological fibrocartilaginous tissues from the goat, including the menisci and the enthesis, were also positive for both laminin and collagen type IV pericellularly. In degenerated fibrocartilage, including intervertebral disc, as in degenerated hyaline cartilage only collagen type IV was found pericellularly around chondrocytes but with less intense staining than in non-degenerated tissue. In calcified cartilage, some cells were positive for laminin but not type IV collagen. We report differences in expression of the BM molecules, laminin and collagen type IV, in normal and degenerative cartilaginous tissues from adult humans and goats. In degenerative tissues laminin is depleted from the pericellular matrix before collagen type IV. The findings may inform future studies of the processes underlying cartilage degeneration and the functional roles of these 2 extracellular matrix proteins

  9. Distribution of Basement Membrane Molecules, Laminin and Collagen Type IV, in Normal and Degenerated Cartilage Tissues

    Science.gov (United States)

    Toh, Wei Seong; Gomoll, Andreas H.; Olsen, Bjørn Reino; Spector, Myron

    2014-01-01

    Objective: The objective of the present study was to investigate the presence and distribution of 2 basement membrane (BM) molecules, laminin and collagen type IV, in healthy and degenerative cartilage tissues. Design: Normal and degenerated tissues were obtained from goats and humans, including articular knee cartilage, the intervertebral disc, and meniscus. Normal tissue was also obtained from patella-tibial enthesis in goats. Immunohistochemical analysis was performed using anti-laminin and anti–collagen type IV antibodies. Human and goat skin were used as positive controls. The percentage of cells displaying the pericellular presence of the protein was graded semiquantitatively. Results: When present, laminin and collagen type IV were exclusively found in the pericellular matrix, and in a discrete layer on the articulating surface of normal articular cartilage. In normal articular (hyaline) cartilage in the human and goat, the proteins were found co-localized pericellularly. In contrast, in human osteoarthritic articular cartilage, collagen type IV but not laminin was found in the pericellular region. Nonpathological fibrocartilaginous tissues from the goat, including the menisci and the enthesis, were also positive for both laminin and collagen type IV pericellularly. In degenerated fibrocartilage, including intervertebral disc, as in degenerated hyaline cartilage only collagen type IV was found pericellularly around chondrocytes but with less intense staining than in non-degenerated tissue. In calcified cartilage, some cells were positive for laminin but not type IV collagen. Conclusions: We report differences in expression of the BM molecules, laminin and collagen type IV, in normal and degenerative cartilaginous tissues from adult humans and goats. In degenerative tissues laminin is depleted from the pericellular matrix before collagen type IV. The findings may inform future studies of the processes underlying cartilage degeneration and the functional

  10. Tissue banking for management of nuclear casualties

    International Nuclear Information System (INIS)

    Singh, Rita

    2014-01-01

    The proliferation of nuclear material and technology has made the acquisition and adversarial use more probable than ever. Devastating medical consequences would follow a nuclear detonation due to the thermal, blast and radiation effects of the weapon. Atomic explosions at Hiroshima and Nagasaki demonstrated the human agonies on vast scale. A full range of medical modalities are required to decrease the morbidity and mortality as a result of the use of nuclear weapons. Biological tissues from human donor like bone, skin, amniotic membrane and other soft tissues can be used for repair or reconstruction of the injured part of the body. Tissues from human donor can be processed and banked for orthopaedic, spinal, trauma and other surgical procedures. Processed tissues can be provided by the tissue banks and can be of great assistance in the treatment of injuries due to the nuclear weapon. The use of allograft tissue avoids the donor site morbidity and reduces the operating time, expense and trauma associated with the acquisition of autografts. Further, allografts have the added advantage of being available in large quantities. This has led to a global increase in allogeneic transplantation and development of tissue banking. The aim of the tissue bank is to provide a wide range of processed biological tissues free from any transmissible disease, that help to restore the growth and function of the damaged tissues. Skin dressings or skin substitutes like allograft skin, xenograft skin and amniotic membrane can be used for the treatment of thermal burns and radiation induced skin injuries. Bone allografts can be used for reconstructive approaches to the skeletal system. Tissue banking would thus ensure health care to the military personnel and population following a nuclear detonation. (author)

  11. Membrane Vesicles of Group B Streptococcus Disrupt Feto-Maternal Barrier Leading to Preterm Birth.

    Directory of Open Access Journals (Sweden)

    Manalee Vishnu Surve

    2016-09-01

    Full Text Available Infection of the genitourinary tract with Group B Streptococcus (GBS, an opportunistic gram positive pathogen, is associated with premature rupture of amniotic membrane and preterm birth. In this work, we demonstrate that GBS produces membrane vesicles (MVs in a serotype independent manner. These MVs are loaded with virulence factors including extracellular matrix degrading proteases and pore forming toxins. Mice chorio-decidual membranes challenged with MVs ex vivo resulted in extensive collagen degradation leading to loss of stiffness and mechanical weakening. MVs when instilled vaginally are capable of anterograde transport in mouse reproductive tract. Intra-amniotic injections of GBS MVs in mice led to upregulation of pro-inflammatory cytokines and inflammation mimicking features of chorio-amnionitis; it also led to apoptosis in the chorio-decidual tissue. Instillation of MVs in the amniotic sac also resulted in intrauterine fetal death and preterm delivery. Our findings suggest that GBS MVs can independently orchestrate events at the feto-maternal interface causing chorio-amnionitis and membrane damage leading to preterm birth or fetal death.

  12. Membrane Vesicles of Group B Streptococcus Disrupt Feto-Maternal Barrier Leading to Preterm Birth

    Science.gov (United States)

    Sthanam, Lakshmi Kavitha; Srivastava, Rohit; Basu, Bhakti; Dutta, Suryendu; Sen, Shamik; Modi, Deepak

    2016-01-01

    Infection of the genitourinary tract with Group B Streptococcus (GBS), an opportunistic gram positive pathogen, is associated with premature rupture of amniotic membrane and preterm birth. In this work, we demonstrate that GBS produces membrane vesicles (MVs) in a serotype independent manner. These MVs are loaded with virulence factors including extracellular matrix degrading proteases and pore forming toxins. Mice chorio-decidual membranes challenged with MVs ex vivo resulted in extensive collagen degradation leading to loss of stiffness and mechanical weakening. MVs when instilled vaginally are capable of anterograde transport in mouse reproductive tract. Intra-amniotic injections of GBS MVs in mice led to upregulation of pro-inflammatory cytokines and inflammation mimicking features of chorio-amnionitis; it also led to apoptosis in the chorio-decidual tissue. Instillation of MVs in the amniotic sac also resulted in intrauterine fetal death and preterm delivery. Our findings suggest that GBS MVs can independently orchestrate events at the feto-maternal interface causing chorio-amnionitis and membrane damage leading to preterm birth or fetal death. PMID:27583406

  13. Cell Membrane Tracking in Living Brain Tissue Using Differential Interference Contrast Microscopy.

    Science.gov (United States)

    Lee, John; Kolb, Ilya; Forest, Craig R; Rozell, Christopher J

    2018-04-01

    Differential interference contrast (DIC) microscopy is widely used for observing unstained biological samples that are otherwise optically transparent. Combining this optical technique with machine vision could enable the automation of many life science experiments; however, identifying relevant features under DIC is challenging. In particular, precise tracking of cell boundaries in a thick ( ) slice of tissue has not previously been accomplished. We present a novel deconvolution algorithm that achieves the state-of-the-art performance at identifying and tracking these membrane locations. Our proposed algorithm is formulated as a regularized least squares optimization that incorporates a filtering mechanism to handle organic tissue interference and a robust edge-sparsity regularizer that integrates dynamic edge tracking capabilities. As a secondary contribution, this paper also describes new community infrastructure in the form of a MATLAB toolbox for accurately simulating DIC microscopy images of in vitro brain slices. Building on existing DIC optics modeling, our simulation framework additionally contributes an accurate representation of interference from organic tissue, neuronal cell-shapes, and tissue motion due to the action of the pipette. This simulator allows us to better understand the image statistics (to improve algorithms), as well as quantitatively test cell segmentation and tracking algorithms in scenarios, where ground truth data is fully known.

  14. Treatment of periodontal disease with guided tissue regeneration technique using a hydroxyapatite and polycaprolactone membrane

    Directory of Open Access Journals (Sweden)

    L.M.A. Martins

    Full Text Available ABSTRACT The aim of this study was to evaluate the use of a malleable membrane composed of hydroxyapatite (60% and polycaprolactone (40% as treatment of periodontal disease experimentally induced in dogs. A bone defect of standardized dimensions was created between the roots of the third and fourth premolar of 12 dogs for periodontal disease induction. Six dogs had the defect covered by the membrane and six dogs received only standard treatment for periodontal disease, also applied to dogs in the treated group. The animals were clinically monitored during the experiment. Radiographs were taken after surgery and at 60 days after treatment initiation. Clinical attachment level was also assessed in those moments. On the 60th day, dental sample of all animals, containing tooth, defect and periodontal tissues, were harvested, fixed in formalin and analyzed by microtomography and histology. During the experimental period, the animals showed no pain and purulent discharge, however, there was dehiscence in 50% of animals and membrane exposure in five out of six animals in the treated group. Clinical attachment level showed no difference between groups. Radiographs showed radiopacity equal to the alveolar bone in both groups. The microtomography revealed that the control group had higher bone volume in the defect compared to the treated group; however, the furcation was not filled by new alveolar bone in any animal. Histological analysis revealed that junctional epithelium invasion was lighter in the control group. New bone was only observed in the apical edge of the defect in both groups. Although the composite is biocompatible and able to keep the space of the defect, it did not promote periodontal tissue regeneration within 60 days of observation.

  15. Proteomics of plasma membranes from poplar trees reveals tissue distribution of transporters, receptors, and proteins in cell wall formation.

    Science.gov (United States)

    Nilsson, Robert; Bernfur, Katja; Gustavsson, Niklas; Bygdell, Joakim; Wingsle, Gunnar; Larsson, Christer

    2010-02-01

    By exploiting the abundant tissues available from Populus trees, 3-4 m high, we have been able to isolate plasma membranes of high purity from leaves, xylem, and cambium/phloem at a time (4 weeks after bud break) when photosynthesis in the leaves and wood formation in the xylem should have reached a steady state. More than 40% of the 956 proteins identified were found in the plasma membranes of all three tissues and may be classified as "housekeeping" proteins, a typical example being P-type H(+)-ATPases. Among the 213 proteins predicted to be integral membrane proteins, transporters constitute the largest class (41%) followed by receptors (14%) and proteins involved in cell wall and carbohydrate metabolism (8%) and membrane trafficking (8%). ATP-binding cassette transporters (all members of subfamilies B, C, and G) and receptor-like kinases (four subfamilies) were two of the largest protein families found, and the members of these two families showed pronounced tissue distribution. Leaf plasma membranes were characterized by a very high proportion of transporters, constituting almost half of the integral proteins. Proteins involved in cell wall synthesis (such as cellulose and sucrose synthases) and membrane trafficking were most abundant in xylem plasma membranes in agreement with the role of the xylem in wood formation. Twenty-five integral proteins and 83 soluble proteins were exclusively found in xylem plasma membranes, which identifies new candidates associated with cell wall synthesis and wood formation. Among the proteins uniquely found in xylem plasma membranes were most of the enzymes involved in lignin biosynthesis, which suggests that they may exist as a complex linked to the plasma membrane.

  16. Concise Review: Amniotic Fluid Stem Cells: The Known, the Unknown, and Potential Regenerative Medicine Applications.

    Science.gov (United States)

    Loukogeorgakis, Stavros P; De Coppi, Paolo

    2017-07-01

    The amniotic fluid has been identified as an untapped source of cells with broad potential, which possess immunomodulatory properties and do not have the ethical and legal limitations of embryonic stem cells. CD117(c-Kit)+ cells selected from amniotic fluid have been shown to differentiate into cell lineages representing all three embryonic germ layers without generating tumors, making them ideal candidates for regenerative medicine applications. Moreover, their ability to engraft in injured organs and modulate immune and repair responses of host tissues, suggest that transplantation of such cells may be useful for the treatment of various degenerative and inflammatory diseases. Although significant questions remain regarding the origin, heterogeneous phenotype, and expansion potential of amniotic fluid stem cells, evidence to date supports their potential role as a valuable stem cell source for the field of regenerative medicine. Stem Cells 2017;35:1663-1673. © 2016 AlphaMed Press.

  17. Radiation processing of biological tissues for nuclear disaster management

    International Nuclear Information System (INIS)

    Singh, Rita

    2012-01-01

    A number of surgical procedures require tissue substitutes to repair or replace damaged or diseased tissues. Biological tissues from human donor like bone, skin, amniotic membrane and other soft tissues can be used for repair or reconstruction of the injured part of the body. Tissues from human donor can be processed and banked for orthopaedic, spinal, trauma and other surgical procedures. Allograft tissues provide an excellent alternative to autografts. The use of allograft tissue avoids the donor site morbidity and reduces the operating time, expense and trauma associated with the acquisition of autografts. Further, allografts have the added advantage of being available in large quantities. This has led to a global increase in allogeneic transplantation and development of tissue banking. However, the risk of infectious disease transmission via tissue allografts is a major concern. Therefore, tissue allografts should be sterilized to make them safe for clinical use. Radiation processing has well appreciated technological advantages and is the most suitable method for sterilization of biological tissues. Radiation processed biological tissues can be provided by the tissue banks for the management of injuries due to a nuclear disaster. A nuclear detonation will result in a large number of casualties due to the heat, blast and radiation effects of the weapon. Skin dressings or skin substitutes like allograft skin, xenograft skin and amniotic membrane can be used for the treatment of thermal burns and radiation induced skin injuries. Bone grafts can be employed for repairing fracture defects, filling in destroyed regions of bone, management of open fractures and joint injuries. Radiation processed tissues have the potential to repair or reconstruct damaged tissues and can be of great assistance in the treatment of injuries due to the nuclear weapon. (author)

  18. The correlation between histologic placentitis and amnionitis and the amnioniotic fluid's inflammatory cytokines in case of spontaneous pre-term labor with intact membrane

    Directory of Open Access Journals (Sweden)

    Agus Abadi

    2001-12-01

    Full Text Available Pre-term labor is presumed to result from spreading of lower genital infection to upper part, subsequently to decidual and choioamniotic tissues. Host response to this injury include the expression of protein which is responsible to the inflammatory reactions. The expression of the inflammatory cytokines such as IL-1β, IL-6, IL-8 and TNF-α increase in case of infection.These cytokines may play an essential role in the pathophysiology of spontaneous pretem labor with intact membrane.An observational analytic cohort study was caried out on cases of spontaneous pre-tefln labor with intact membrane. The objectives of this study are to examine the relationship between l the histologic amnionitis and placentitis and the incidence of preterm delivery,2 the expression of amniotic fluid's IL-1β, IL-6, IL-8 and TNF-α and the incidence of preterm delivery, 3 the level of amniotic fluid's IL-1β, IL-6, IL-8 and TNF-α and the grade of histologic amnionitis and placentitis in case of pre-term labor with intact membrane. Cases of spontaneous Pre'teftn labor with intact membrane which underwent transabdominal amniocentesis at admission and managed as standard procedure for pre-term labor with intact membrane. Atl of the cases were observed until the delivery of the baby, eithir preterm or term. The membrane and the placentawere cut postnatally and then the histologic acute inflammation eyaluated based on the criteria of Salafia.The level of amniotic fluid IL-1β, IL-6, IL-8 and TNF-α were analyzed quantitatively by Elisa method. This study showed thet the degree of histologic amnionitis and placentitis, and the level of amniotic fluid's IL-1β, IL-6, IL-8 and TNF-α were significantly higher in pre-term compared to terrn deliveries (p<0.05 and lhere were a positive correlation between the grade of histoLogic inflammation and the level of amniotic fluid's cytokines (Spearmann Rank Conelation test; p<0,05 in cases of preterm labor with intact membrane. The

  19. Tissue integrity is essential for ectopic implantation of human endometrium in the chicken chorioallantoic membrane.

    Science.gov (United States)

    Nap, Annemiek W; Groothuis, Patrick G; Demir, Ayse Y; Maas, Jacques W M; Dunselman, Gerard A J; de Goeij, Anton F P M; Evers, Johannes L H

    2003-01-01

    Not all women with patent tubes develop clinically manifest endometriosis. Quality and quantity of endometrium in retrograde menstruation may be the determining factor in the development of the disease. We hypothesize that retrograde shedding of endometrial fragments with preserved integrity facilitates implantation of endometrium in ectopic locations, resulting in endometriotic lesion development. We evaluate the impact of tissue integrity on the success of endometriosis-like lesion development in the chicken embryo chorioallantoic membrane (CAM) model. Menstrual and non-menstrual (cyclic) endometrium were collected by biopsy, and either minced or enzymatically dispersed. Spontaneously shed menstrual effluent was collected by a menstrual cup, and cells and tissue were isolated. We evaluated whether infiltration or lesion formation in the CAM occurred after transplantation of endometrium onto the CAM. Transplantation of biopsied menstrual and cyclic endometrium fragments, and of endometrium fragments >1 mm(3) isolated from menstrual effluent, resulted in lesion formation. Transplantation of endometrial cells isolated from menstrual effluent did not lead to lesion formation. After transplantation of digested biopsied cyclic endometrium, infiltration in the CAM but no lesions were observed. In the CAM assay, integrity of tissue architecture determines success of implantation of human endometrium in ectopic locations.

  20. Transplante de células-tronco epiteliais límbicas alógenas expandidas ex vivo sobre membrana amniótica: relato de caso Transplantation of allogenic limbal epithelial stem cells cultivated ex vivo on amniotic membrane: case report

    Directory of Open Access Journals (Sweden)

    José Álvaro Pereira Gomes

    2009-04-01

    Full Text Available Paciente apresentou falência de transplante de limbo e conjuntiva de doador vivo alógeno no olho direito após ceratoconjuntivite epidêmica. Após alguns meses, foi submetida a transplante de células-tronco epiteliais límbicas alógenas cultivadas ex vivo sobre membrana amniótica (primeiro caso no Brasil, tendo evoluído com epitelização total da córnea e melhora da acuidade visual. Após o 3º mês da cirurgia, iniciou-se neovascularização superficial periférica com piora da transparência corneana. A visão manteve-se 0,1 após um ano de cirurgia.Case report of a patient who developed failure of an allogenic living related conjunctival limbal transplantation in the right eye after an episode of epidemic keratoconjunctivitis. After a few months, she underwent transplantation of allogenic limbal epithelial stem cells cultivated ex vivo on amniotic membrane (first case in Brazil. The patient evolved with total corneal epithelialization and improvement of the visual acuity. Three months after the surgery, peripheral superficial neovascularization with worsening of the corneal transparency was observed. The vision remained 0.1 after one year of the transplantation.

  1. Bioactive and metal uptake studies of carboxymethyl chitosan-graft-D-glucuronic acid membranes for tissue engineering and environmental applications.

    Science.gov (United States)

    Jayakumar, R; Rajkumar, M; Freitas, H; Sudheesh Kumar, P T; Nair, S V; Furuike, T; Tamura, H

    2009-08-01

    Carboxymethyl chitosan-graft-D-glucuronic acid (CMCS-g-D-GA) was prepared by grafting D-GA onto CMCS in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and then the membranes were made from it. In this work, the bioactivity studies of CMCS-g-D-GA membranes were carried out and then characterized by SEM, CLSM, XRD and FT-IR. The CMCS-g-D-GA membranes were found to be bioactive. The adsorption of Ni2+, Zn2+ and Cu2+ ions onto CMCS-g-D-GA membranes has also been investigated. The maximum adsorption capacity of CMCS-g-D-GA for Ni2+, Zn2+ and Cu2+ was found to be 57, 56.4 and 70.2 mg/g, respectively. Hence, these membranes were useful for tissue engineering, environmental and water purification applications.

  2. Acid-extrusion from tissue: the interplay between membrane transporters and pH buffers.

    Science.gov (United States)

    Hulikova, Alzbeta; Harris, Adrian L; Vaughan-Jones, Richard D; Swietach, Pawel

    2012-01-01

    The acid-base balance of cells is related to the concentration of free H⁺ ions. These are highly reactive, and their intracellular concentration must be regulated to avoid detrimental effects to the cell. H⁺ ion dynamics are influenced by binding to chelator substances ('buffering'), and by the production, diffusion and membrane-transport of free H⁺ ions or of the H⁺-bound chelators. Intracellular pH (pHi) regulation aims to balance this system of diffusion-reaction-transport processes at a favourable steady-state pHi. The ability of cells to regulate pHi may set a limit to tissue growth and can be subject to selection pressures. Cancer cells have been postulated to respond favourably to such selection pressures by evolving a better means of pHi regulation. A particularly important feature of tumour pHi regulation is acid-extrusion, which involves H⁺-extrusion and HCO₃⁻-uptake by membrane-bound transporter-proteins. Extracellular CO₂/HCO₃⁻ buffer facilitates these membrane-transport processes. As a mobile pH-buffer, CO₂/HCO₃⁻ protects the extracellular space from excessive acidification that could otherwise inhibit further acid-extrusion. CO₂/HCO₃⁻ also provides substrate for HCO₃⁻-transporters. However, the inherently slow reaction kinetics of CO₂/HCO₃⁻ can be rate-limiting for acid-extrusion. To circumvent this, cells can express extracellular-facing carbonic anhydrase enzymes to accelerate the attainment of equilibrium between CO₂, HCO₃⁻ and H⁺. The acid-extrusion apparatus has been proposed as a target for anti-cancer therapy. The major targets include H⁺ pumps, Na⁺/H⁺ exchangers and carbonic anhydrases. The effectiveness of such therapy will depend on the correct identification of rate-limiting steps in pHi regulation in a specific type of cancer.

  3. Neutrophil Extracellular Traps in the Amniotic Cavity of Women with Intra-Amniotic Infection: A New Mechanism of Host Defense.

    Science.gov (United States)

    Gomez-Lopez, Nardhy; Romero, Roberto; Xu, Yi; Miller, Derek; Unkel, Ronald; Shaman, Majid; Jacques, Suzanne M; Panaitescu, Bogdan; Garcia-Flores, Valeria; Hassan, Sonia S

    2017-08-01

    Neutrophil extracellular traps (NETs) control microbial infections through their antimicrobial activities attributed to DNA, histones, granules, and cytoplasmic proteins (eg, elastase). Intra-amniotic infection is characterized by the influx of neutrophils into the amniotic cavity; therefore, the aim of this study was to determine whether amniotic fluid neutrophils form NETs in this inflammatory process. Amniotic fluid samples from women with intra-amniotic infection (n = 15) were stained for bacteria detection using fluorescent dyes. Amniotic fluid neutrophils were purified by filtration. As controls, neutrophils from maternal blood samples (n = 3) were isolated by density gradients. Isolated neutrophils were plated onto glass cover slips for culture with and without 100 nM of phorbol-12-myristate-13-acetate (PMA). NET formation was assessed by 4',6-diamidino-2-phenylindole (DAPI) staining and scanning electron microscopy. Different stages of NET formation were visualized using antibodies against elastase and histone H3, in combination with DAPI staining, by confocal microscopy. Finally, maternal or neonatal neutrophils were added to amniotic fluid samples from women without intra-amniotic infection (n = 4), and NET formation was evaluated by DAPI staining. (1) NETs were present in the amniotic fluid of women with intra-amniotic infection; (2) all of the amniotic fluid samples had detectable live and dead bacteria associated with the presence of NETs; (3) in contrast to neutrophils from the maternal circulation, amniotic fluid neutrophils did not require PMA stimulation to form NETs; (4) different stages of NET formation were observed by co-localizing elastase, histone H3, and DNA in amniotic fluid neutrophils; and (5) neither maternal nor neonatal neutrophils form NETs in the amniotic fluid of women without intra-amniotic infection. NETs are detectable in the amniotic fluid of women with intra-amniotic infection.

  4. A Solvent-Free Surface Suspension Melt Technique for Making Biodegradable PCL Membrane Scaffolds for Tissue Engineering Applications

    Directory of Open Access Journals (Sweden)

    Ratima Suntornnond

    2016-03-01

    Full Text Available In tissue engineering, there is limited availability of a simple, fast and solvent-free process for fabricating micro-porous thin membrane scaffolds. This paper presents the first report of a novel surface suspension melt technique to fabricate a micro-porous thin membrane scaffolds without using any organic solvent. Briefly, a layer of polycaprolactone (PCL particles is directly spread on top of water in the form of a suspension. After that, with the use of heat, the powder layer is transformed into a melted layer, and following cooling, a thin membrane is obtained. Two different sizes of PCL powder particles (100 µm and 500 µm are used. Results show that membranes made from 100 µm powders have lower thickness, smaller pore size, smoother surface, higher value of stiffness but lower ultimate tensile load compared to membranes made from 500 µm powder. C2C12 cell culture results indicate that the membrane supports cell growth and differentiation. Thus, this novel membrane generation method holds great promise for tissue engineering.

  5. Characterizing nanoscale topography of the aortic heart valve basement membrane for tissue engineering heart valve scaffold design.

    Science.gov (United States)

    Brody, Sarah; Anilkumar, Thapasimuthu; Liliensiek, Sara; Last, Julie A; Murphy, Christopher J; Pandit, Abhay

    2006-02-01

    A fully effective prosthetic heart valve has not yet been developed. A successful tissue-engineered valve prosthetic must contain a scaffold that fully supports valve endothelial cell function. Recently, topographic features of scaffolds have been shown to influence the behavior of a variety of cell types and should be considered in rational scaffold design and fabrication. The basement membrane of the aortic valve endothelium provides important parameters for tissue engineering scaffold design. This study presents a quantitative characterization of the topographic features of the native aortic valve endothelial basement membrane; topographical features were measured, and quantitative data were generated using scanning electron microscopy (SEM), atomic force microscopy (AFM), transmission electron microscopy (TEM), and light microscopy. Optimal conditions for basement membrane isolation were established. Histological, immunohistochemical, and TEM analyses following decellularization confirmed basement membrane integrity. SEM and AFM photomicrographs of isolated basement membrane were captured and quantitatively analyzed. The basement membrane of the aortic valve has a rich, felt-like, 3-D nanoscale topography, consisting of pores, fibers, and elevations. All features measured were in the sub-100 nm range. No statistical difference was found between the fibrosal and ventricular surfaces of the cusp. These data provide a rational starting point for the design of extracellular scaffolds with nanoscale topographic features that mimic those found in the native aortic heart valve basement membrane.

  6. Role of Echogenic Amniotic Fluid Particles and Optical Density in ...

    African Journals Online (AJOL)

    This study was aimed to correlate echogenic amniotic fluid particle size (AFPS) in late third trimester to fetal lung maturity and amniotic fluid optical density (AFOD) at labor. AFPS were measured with specified criteria by real time transabdominal USG (3.5MHz) while Amniotic Fluid Index (AFI) was measured during routine ...

  7. Amniotic Fluid Soluble Myeloid Differentiation-2 (sMD-2) as Regulator of Intra-amniotic Inflammation in Infection-induced Preterm Birth.

    Science.gov (United States)

    Dulay, Antonette T; Buhimschi, Catalin S; Zhao, Guomao; Oliver, Emily A; Abdel-Razeq, Sonya S; Shook, Lydia L; Bahtiyar, Mert O; Buhimschi, Irina A

    2015-06-01

    TLR4 mediates host responses to pathogens through a mechanism that involves protein myeloid differentiation-2 (MD-2) and its soluble form sMD-2. The role of sMD2 in intra-amniotic inflammation-induced preterm birth has not been previously explored. Human amniotic fluid (AF) sMD-2 was studied by Western blotting in 152 AF samples of patients who had an amniocentesis to rule-out infection (yes infection, n = 50; no infection, n = 50) or women with normal pregnancy outcome (second trimester genetic karyotyping, n = 26; third trimester lung maturity testing, n = 26). Histological localization and mRNA expression of MD2 in fetal membranes were studied by immunohistochemistry and RT-PCR. The ability of fetal membrane to release sMD-2 and inflammatory cytokines was studied in vitro. Human AF contains three sMD-2 proteoforms whose levels of expression were lower at term. Intra-amniotic infection upregulated sMD-2. MD-2 mRNA and immunohistochemistry findings concurred. In vitro, LPS and monensin increased, while cycloheximide decreased sMD-2 production. Recombinant sMD-2 modulated TNF-α and IL-6 levels in a dose- and time-dependent fashion. sMD2 proteoforms are constitutively present in human AF. The intensity of the intra-amniotic inflammatory response to bacteria or perhaps to other TLR4 ligands may be facilitated through synthesis and release of sMD2 by the amniochorion. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  8. [Morphology of basement membrane and associated matrix proteins in normal and pathological tissues].

    Science.gov (United States)

    Nerlich, A

    1995-01-01

    Basement membranes (BM) are specialized structures of the extracellular matrix. Their composition is of particular importance for the maintenance of normal morphological and functional properties of a multitude of organs and tissue systems and it is thus required for regular homeostasis of body function. Generally, they possess three main functions, i.e. participation in the maintenance of tissue structure, control of fluid and substrate exchange, and regulation of cell growth and differentiation. BMs are made up by various components which are in part specifically localized within the BM zone, or which represent ubiquitous matrix constituents with specific quantitative and/or qualitative differences in their localization. On the basis of a thorough immunohistochemical analysis of normal and diseased tissues, we provide here a concept of "functional morphology/pathomorphology" of the different BM components analyzed: 1.) The ubiquitous BM-constituent collagen IV primarily stabilizes the BM-zone and thus represents the "backbone" of the BM providing mechanical strength. Its loss leads to cystic tissue transformation as it is evidenced from the analysis of polycystic nephropathies. Thus, in other cystic tissue transformations a similar formal pathogenesis may be present. 2.) The specific localization of collagen VII as the main structural component of anchoring fibrils underlines the mechanical anchoring function of this collagenous protein. Defects in this protein lead to hereditary epidermolysis. The rapid re-occurrence of epidermal collagen VII during normal human wound healing indicates a quick reconstitution of the mechanical tensile strength of healing wounds. 3.) The BM-specific heparan sulfate proteoglycan (HSPG, Perlecan) with its highly negative anionic charge can be assumed to exert filter control. This assumption is corroborated by the localizatory findings of a preferential deposition of HSPG in endothelial and particularly in glomerular BM. Similarly

  9. Amniotic amputation | Ayadi | Pan African Medical Journal

    African Journals Online (AJOL)

    Amniotic band syndrome (ABS) is an uncommon, congenital fetal abnormality. Lower extremity limb defects are the common manifestations of ABS. The most common features include congenital distal ring constrictions, intrauterine amputations, and acrosyndactyly. Rare cases of craniofacial and visceral defects were ...

  10. Characterization of an apically derived epithelial membrane glycoprotein from bovine milk, which is expressed in capillary endothelia in diverse tissues.

    Science.gov (United States)

    Greenwalt, D E; Mather, I H

    1985-02-01

    A glycoprotein (PAS IV) of apparent Mr 76,000 was purified from bovine milk-fat-globule membrane and partially characterized. PAS IV contained mannose, galactose, and sialic acid as principal sugars (approximately 5.3% total carbohydrate [wt/wt]) and existed in milk in at least four isoelectric variants. The glycoprotein appeared to be an integral membrane protein by several criteria. PAS IV was recovered in the detergent phase of Triton X-114 extracts of milk-fat-globule membrane at room temperature. When bound to membrane, PAS IV was resistant to digestion by a number of proteinases, although after solubilization with non-ionic detergents, the protein was readily degraded. Amino acid analysis of the purified protein revealed a high percentage of amino acids with nonpolar residues. The location of PAS IV was determined in bovine tissues by using immunofluorescence techniques. In mammary tissue, PAS IV was located on both the apical surfaces of secretory epithelial cells and endothelial cells of capillaries. This glycoprotein was also detected in endothelial cells of heart, liver, spleen, pancreas, salivary gland, and small intestine. In addition to mammary epithelial cells, PAS IV was also located in certain other epithelial cells, most notably the bronchiolar epithelial cells of lung. The potential usefulness of this protein as a specific marker of capillary endothelial cells in certain tissues is discussed.

  11. Amniotic Fluid Cells Show Higher Pluripotency-Related Gene Expression Than Allantoic Fluid Cells.

    Science.gov (United States)

    Kehl, Debora; Generali, Melanie; Görtz, Sabrina; Geering, Diego; Slamecka, Jaroslav; Hoerstrup, Simon P; Bleul, Ulrich; Weber, Benedikt

    2017-10-01

    Amniotic fluid represents an abundant source of multipotent stem cells, referred as broadly multipotent given their differentiation potential and expression of pluripotency-related genes. However, the origin of this broadly multipotent cellular fraction is not fully understood. Several sources have been proposed so far, including embryonic and extraembryonic tissues. In this regard, the ovine developmental model uniquely allows for direct comparison of fetal fluid-derived cells from two separate fetal fluid cavities, the allantois and the amnion, over the entire duration of gestation. As allantoic fluid mainly collects fetal urine, cells originating from the efferent urinary tract can directly be compared with cells deriving from the extraembryonic amniotic tissues and the fetus. This study shows isolation of cells from the amniotic [ovine amniotic fluid cells (oAFCs)] and allantoic fluid [ovine allantoic fluid cells (oALCs)] in a strictly paired fashion with oAFCs and oALCs derived from the same fetus. Both cell types showed cellular phenotypes comparable to standard mesenchymal stem cells (MSCs), with trilineage differentiation potential, and expression of common ovine MSC markers. However, the expression of MSC markers per single cell was higher in oAFCs as measured by flow cytometry. oAFCs exhibited higher proliferative capacities and showed significantly higher expression of pluripotency-related genes OCT4, STAT3, NANOG, and REX1 by quantitative real-time polymerase chain reaction compared with paired oALCs. No significant decrease of pluripotency-related gene expression was noted over gestation, implying that cells with high differentiation potential may be isolated at the end of pregnancy. In conclusion, this study suggests that cells with highest stem cell characteristics may originate from the fetus itself or the amniotic fetal adnexa rather than from the efferent urinary tract or the allantoic fetal adnexa.

  12. Overview of Twenty Years of Radiation and Tissue Banking Activity in Argentina

    International Nuclear Information System (INIS)

    Kairiyama, E.

    2015-01-01

    Radiation sterilization of human tissues in Argentina was a consequence of health care products sterilization by gamma radiation. Radiation technology was implemented in 1970 when the first multipurpose gamma facility was built at the Ezeiza Atomic Centre of CNEA. Organ and tissue transplantation is a well established effective therapy that saves lives and significantly improves the quality of life. Ionizing radiation is used for sterilization in order to provide clinically safe tissue for therapeutic purposes of implantation in every patient in need. Argentina radiation and tissue banking activity started in 1993 with the establishment of two tissue banks using radiation under the IAEA programme of technical cooperation, a skin bank and a bone one. Additionally to this start, other tissue banks have adopted tissue sterilization by irradiation. The compatible tissues sterilized with this methodology are mainly skin (frozen, glycerolized), bone (lyophilized, frozen), and amniotic membrane (glycerolized, frozen, dehydrated). The donation and transplant of human organ, tissue and cells is regulated and coordinated by the National Institute Unique Central Coordinator of Ablation and Implant (INCUCAI). In regards to radiation and nuclear safety, physical protection and nuclear non-proliferation issues are regulated and contorted by the Nuclear Regulatory Authority (ARN). Eight tissue banks use gamma radiation for sterilization of human tissues (6 musculoskeletal, 1 skin and 1 amniotic membrane). Argentina has participated actively in several IAEA projects regarding radiation and tissue banking program, and it has been selected by the IAEA to host the Regional Training Centre for the Latin American region. The following activities were implemented: regional training courses in Buenos Aires, face to face (five) and virtual (four) modalities; collaboration on several materials related to tissue banking and radiation sterilization of tissue allograft, codes of practice for

  13. Surgical membranes as directional delivery devices to generate tissue: testing in an ovine critical sized defect model.

    Directory of Open Access Journals (Sweden)

    Melissa L Knothe Tate

    Full Text Available Pluripotent cells residing in the periosteum, a bi-layered membrane enveloping all bones, exhibit a remarkable regenerative capacity to fill in critical sized defects of the ovine femur within two weeks of treatment. Harnessing the regenerative power of the periosteum appears to be limited only by the amount of healthy periosteum available. Here we use a substitute periosteum, a delivery device cum implant, to test the hypothesis that directional delivery of endogenous periosteal factors enhances bone defect healing.Newly adapted surgical protocols were used to create critical sized, middiaphyseal femur defects in four groups of five skeletally mature Swiss alpine sheep. Each group was treated using a periosteum substitute for the controlled addition of periosteal factors including the presence of collagen in the periosteum (Group 1, periosteum derived cells (Group 2, and autogenic periosteal strips (Group 3. Control group animals were treated with an isotropic elastomer membrane alone. We hypothesized that periosteal substitute membranes incorporating the most periosteal factors would show superior defect infilling compared to substitute membranes integrating fewer factors (i.e. Group 3>Group 2>Group 1>Control.Based on micro-computed tomography data, bone defects enveloped by substitute periosteum enabling directional delivery of periosteal factors exhibit superior bony bridging compared to those sheathed with isotropic membrane controls (Group 3>Group 2>Group 1, Control. Quantitative histological analysis shows significantly increased de novo tissue generation with delivery of periosteal factors, compared to the substitute periosteum containing a collagen membrane alone (Group 1 as well as compared to the isotropic control membrane. Greatest tissue generation and maximal defect bridging was observed when autologous periosteal transplant strips were included in the periosteum substitute.Periosteum-derived cells as well as other factors

  14. Sterilization of biological tissues with ionizing radiation; Esterilizacion de tejidos biologicos con radiacion ionizante

    Energy Technology Data Exchange (ETDEWEB)

    Reyes F, M.L.; Martinez P, M.E.; Luna Z, D. [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)

    1997-07-01

    On June 1994, the National Institute of Nuclear Research (ININ) and the South Central Hospital for High Specialty of PEMEX (HCSAE) began a joint work with the finality to obtain radio sterilized amniotic membranes for to be used as cover (biological bandage) in burnt patients. Subsequently the Chemistry Faculty of UNAM and the National Institute of Cardiology began to collaborate this last with interest on cardiac valves for graft. Starting from 1997, the International Atomic Energy Agency (IAEA) supports this project (MEX/7/008) whose main objective is to set up the basis to establish in Mexico a Radio sterilized Tissue Bank (amniotic membranes, skin, bones, tendons, cardiac valves, etc.) to be used with therapeutic purposes (grafts). The IAEA support has consisted in the equipment acquisition which is fundamental for the Tissue Bank performance such as an experimental irradiator, laminar flow bell, lyophilizer, vacuum sealer and special knives for tissues. Also visits to Mexico of experts have been authorized with the aim of advising to the personnel which participate in the project and scientific visits of this personnel to another tissue banks (Sri Lanka and Argentine). The establishment in Mexico of a Tissue bank will be a great benefit because it will have availability of distinct tissues for grafts and it will reduce the synthetic materials importation which is very expensive. (Author)

  15. Membrane transport properties of equine and macaque ovarian tissues frozen in mixtures of dimethylsulfoxide and ethylene glycol.

    Science.gov (United States)

    Kardak, A; Leibo, S P; Devireddy, R

    2007-10-01

    The rate at which equine and macaque ovarian tissue sections are first cooled from +25 degrees C to +4 degrees C has a significant effect on the measured water transport when the tissues are subsequently frozen in 0.85 M solutions of glycerol, dimethylsulfoxide (DMSO), or ethylene glycol (EG). To determine whether the response of ovarian tissues is altered if they are suspended in mixtures of cryoprotective agents (CPAs), rather than in solutions of a single CPA, we have now measured the subzero water transport from ovarian tissues that were suspended in mixtures of DMSO and EG. Sections of freshly collected equine and macaque ovaries were suspended either in a mixture of 0.9 M EG plus 0.7 M DMSO (equivalent to a mixture of approximately 5% vv of EG and DMSO) or in a 1.6M solution of only DMSO or only EG. The tissue sections were cooled from +25 degrees C to +4 degrees C and then frozen to subzero temperatures at 5 degrees C/min. As the tissues were being frozen, a shape-independent differential scanning calorimeter technique was used to measure water loss from the tissues and, consequently, the best fit membrane permeability parameters (L(pg) and E(Lp)) of ovarian tissues during freezing. In the mixture of DMSO+EG, the respective values of L(pg) and E(Lp) for equine tissue first cooled at 40 degrees C/min between +25 degrees C and +4 degrees C before being frozen were 0.15 microm/min atm and 7.6 kcal/mole. The corresponding L(pg) and E(Lp) values for equine tissue suspended in 1.6M DMSO were 0.12 microm/min atm and 27.2 kcal/mole; in 1.6M EG, the values were 0.06 microm/min atm and 21.9 kcal/mole, respectively. For macaque ovarian tissues suspended in the mixture of DMSO+EG, the respective values of L(pg) and E(Lp) were 0.26 microm/min atm and 26.2 kcal/mole. Similarly, the corresponding L(Lg) and E(Lp) values for macaque tissue suspended in 1.6M DMSO were 0.22 microm/min atm and 31.4 kcal/mole; in 1.6 M EG, the values were 0.20 microm/min atm and 27.9 kcal

  16. Gastric fluid versus amniotic fluid analysis for the identification of intra-amniotic infection due to Ureaplasma species.

    Science.gov (United States)

    Kim, Sun Min; Romero, Roberto; Lee, JoonHo; Chaemsaithong, Piya; Docheva, Nikolina; Yoon, Bo Hyun

    2016-01-01

    Early neonatal sepsis is often due to intra-amniotic infection. The stomach of the neonate contains fluid swallowed before and during delivery. The presence of bacteria as well as neutrophils detected by culture or Gram stain of the gastric fluid during the first day of life is suggestive of exposure to bacteria or inflammation. We undertook this study to determine the relationship between gastric fluid analysis and amniotic fluid obtained by transabdominal amniocentesis in the detection of Ureaplasma species, the most frequent microorganisms responsible for intra-amniotic infection. The study population consisted of 100 singleton pregnant women who delivered preterm neonates (Ureaplasma species was performed. Intra-amniotic inflammation was defined as an elevated amniotic fluid matrix metalloproteinase-8 concentration (>23 ng/mL). (1) Ureaplasma species were detected by culture or PCR in 18% (18/100) of amniotic fluid samples and in 5% (5/100) of gastric fluid samples; (2) among the amniotic fluid cases positive for Ureaplasma species, these microorganisms were identified in 27.8% (5/18) of gastric fluid samples; (3) none of the cases negative for Ureaplasma species in the amniotic fluid were found to be positive for these microorganisms in the gastric fluid; (4) patients with amniotic fluid positive for Ureaplasma species but with gastric fluid negative for these microorganisms had a significantly higher rate of intra-amniotic inflammation, acute histologic chorioamnionitis, and neonatal death than those with both amniotic fluid and gastric fluid negative for Ureaplasma species; and (5) no significant differences were observed in the rate of intra-amniotic inflammation, acute histologic chorioamnionitis, and neonatal death between patients with amniotic fluid positive for Ureaplasma species but with gastric fluid negative for these microorganisms and those with both amniotic fluid and gastric fluid positive for Ureaplasma species. Gastric fluid analysis has 100

  17. Prognostic value of cerebral tissue oxygen saturation during neonatal extracorporeal membrane oxygenation.

    Directory of Open Access Journals (Sweden)

    Marie-Philippine Clair

    Full Text Available Extracorporeal membrane oxygenation support is indicated in severe and refractory respiratory or circulatory failures. Neurological complications are typically represented by acute ischemic or hemorrhagic lesions, which induce higher morbidity and mortality. The primary goal of this study was to assess the prognostic value of cerebral tissue oxygen saturation (StcO2 on mortality in neonates and young infants treated with ECMO. A secondary objective was to evaluate the association between StcO2 and the occurrence of cerebral lesions.This was a prospective study in infants < 3 months of age admitted to a pediatric intensive care unit and requiring ECMO support.The assessment of cerebral perfusion was made by continuous StcO2 monitoring using near-infrared spectroscopy (NIRS sensors placed on the two temporo-parietal regions. Neurological lesions were identified by MRI or transfontanellar echography.Thirty-four infants <3 months of age were included in the study over a period of 18 months. The ECMO duration was 10±7 days. The survival rate was 50% (17/34 patients, and the proportion of brain injuries was 20% (7/34 patients. The mean StcO2 during ECMO in the non-survivors was reduced in both hemispheres (p = 0.0008 right, p = 0.03 left compared to the survivors. StcO2 was also reduced in deceased or brain-injured patients compared to the survivors without brain injury (p = 0.002.StcO2 appears to be a strong prognostic factor of survival and of the presence of cerebral lesions in young infants during ECMO.

  18. Reinforcement of poly-l-lactic acid electrospun membranes with strontium borosilicate bioactive glasses for bone tissue engineering.

    Science.gov (United States)

    Fernandes, João S; Gentile, Piergiorgio; Martins, Margarida; Neves, Nuno M; Miller, Cheryl; Crawford, Aileen; Pires, Ricardo A; Hatton, Paul; Reis, Rui L

    2016-10-15

    Herein, for the first time, we combined poly-l-lactic acid (PLLA) with a strontium borosilicate bioactive glass (BBG-Sr) using electrospinning to fabricate a composite bioactive PLLA membrane loaded with 10% (w/w) of BBG-Sr glass particles (PLLA-BBG-Sr). The composites were characterised by scanning electron microscopy (SEM) and microcomputer tomography (μ-CT), and the results showed that we successfully fabricated smooth and uniform fibres (1-3μm in width) with a homogeneous distribution of BBG-Sr microparticles (bone marrow-derived mesenchymal stem cells (BM-MSCs) demonstrated that PLLA-BBG-Sr membranes promoted the osteogenic differentiation of the cells as demonstrated by increased alkaline phosphatase activity and up-regulated osteogenic gene expression (Alpl, Sp7 and Bglap) in relation to PLLA alone. These results strongly suggest that the composite PLLA membranes reinforced with the BBG-Sr glass particles have potential as an effective biomaterial capable of promoting bone regeneration. PLLA membranes were reinforced with 10% (w/w) of strontium-bioactive borosilicate glass microparticles, and their capacity to induce the osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs) was evaluated. These membranes presented an increased: degradability, water uptake, Young modulus and tensile strength. We also demonstrated that these membranes are non-cytotoxic and promote the attachment of BM-MSCs. The addition of the glass microparticles into the PLLA membranes promoted the increase of ALP activity (under osteogenic conditions), as well as the BM-MSCs osteogenic differentiation as shown by the upregulation of Alpl, Sp7 and Bglap gene expression. Overall, we demonstrated that the reinforcement of PLLA with glass microparticles results in a biomaterial with the appropriate properties for the regeneration of bone tissue. Copyright © 2016 Acta Materialia Inc. All rights reserved.

  19. Amniotic band syndrome: A clinical brief

    Directory of Open Access Journals (Sweden)

    Dasaradha Ramireddy Malireddy

    2017-01-01

    Full Text Available Amniotic band syndrome (ABS results from bands of amnion entangling fetal parts. They may manifest as constriction rings or complex congenital anomalies resulting in stillbirth. Karyotyping is important for exclusion of inherited disorders and proper counseling. Two case reports one stillbirth and the other with constriction ring of fingers and mild hydronephrosis are presented. The aim of this paper is to make awareness and stress the need for doing thorough work-up in all cases of constriction bands.

  20. Anti-inflammatory Elafin in human fetal membranes.

    Science.gov (United States)

    Stalberg, Cecilia; Noda, Nathalia; Polettini, Jossimara; Jacobsson, Bo; Menon, Ramkumar

    2017-02-01

    Elafin is a low molecular weight protein with antileukoproteinase, anti-inflammatory, antibacterial and immunomodulating properties. The profile of Elafin in fetal membranes is not well characterized. This study determined the changes in Elafin expression and concentration in human fetal membrane from patients with preterm prelabor rupture of membranes (PPROM) and in vitro in response to intra-amniotic polymicrobial pathogens. Elafin messenger RNA (mRNA) expressions were studied in fetal membranes from PPROM, normal term as well as in normal term not in labor membranes in an organ explant system treated (24 h) with lipopolysaccharide (LPS), using quantitative reverse transcription-polymerase chain reaction (RT-PCR). Enzyme-linked immunosorbent assay (ELISA) measured Elafin concentrations in culture supernatants from tissues treated with LPS and polybacterial combinations of heat-inactivated Mycoplasma hominis (MH), Ureaplasma urealyticum (UU) and Gardnerella vaginalis (GV). Elafin mRNA expression in fetal membranes from women with PPROM was significantly higher compared to women who delivered at term after normal pregnancy (5.09±3.50 vs. 11.71±2.21; Pmembranes showed a significantly increased Elafin m-RNA expression (Pmembranes also showed no changes in Elafin protein concentrations compared to untreated controls. Higher Elafin expression in PPROM fetal membranes suggests a host response to an inflammatory pathology. However, lack of Elafin response to LPS and polymicrobial treatment is indicative of the minimal anti-inflammatory impact of this molecule in fetal membranes.

  1. Clinical comparison of guided tissue regeneration, with collagen membrane and bone graft, versus connective tissue graft in the treatment of gingival recessions

    Directory of Open Access Journals (Sweden)

    Haghighati F

    2006-06-01

    Full Text Available Background and Aim: Increasing patient demands for esthetic, put the root coverage procedures in particular attention. Periodontal regeneration with GTR based root coverage methods is the most common treatment used. The purpose of this study was to compare guided tissue regeneration (GTR with collagen membrane and a bone graft, with sub-epithelial connective tissue graft (SCTG, in treatment of gingival recession. Materials and Methods: In this randomized clinical trial study, eleven healthy patients with no systemic diseases who had miller’s class I or II recession defects (gingival recession  2mm were treated with SCTG or GTR using a collagen membrane and a bone graft. Clinical measurements were obtained at baseline and 6 months after surgery. These clinical measurements included recession depth (RD, recession width (RW, probing depth (PD, and clinical attachment level (CAL. Data were analyzed using independent t test with p<0.05 as the limit of significance. Results: Both treatment methods resulted in a statistically significant reduction of recession depth (SCTG=2.3mm, GTR=2.1mm; P<0.0001. CAL gain after 6 months was also improved in both groups (SCG= 2.5mm, GTR=2.1mm, compared to baseline (P<0.0001. No statistical differences were observed in RD, RW, CAL between test and control groups. Root coverage was similar in both methods (SCTG= 74.2%, GTR= 62.6%, P=0.87. Conclusion: Based on the results of this study, the two techniques are clinically comparable. Therefore the use of collagen membrane and a bovine derived xenograft may alleviate the need for connective tissue graft.

  2. Amniotic Fluid β2- Microglobulin Measurements

    Directory of Open Access Journals (Sweden)

    Emine Aydın

    2016-02-01

    Full Text Available OBJECTIVE: To determine β2-microglobulin levels in amniotic fluid during the course of second trimester. STUDY DESIGN: One hundred patient’s amniotic fluid β2-microglobulin levels had been evaluated retrospectively (March-October 2009. The most common amniocentesis indication was advanced maternal age (33.3%. Others were; high risk result for triple test (18.5%, high risk result for double test (6.48%, ventriculomegaly (5.55%, obstetric history for fetus with down syndrome (4.62%, the presence of soft markers on ultrasound (13.8%, others (17.8%. Patients average gravida was 2.66 (range: 1-6, parity was 0.75 (range: 0-3, abortion was 0.65 (range: 0-3. RESULTS: All patients were at second trimester and the average gestational week was 17.7 (range 15- 22. Patients were divided into four groups (15th, 16th, 17-18th and 19-20th gestational weeks. We have demonstrated that amniotic fluid β2-microglobulin levels are increased progressively throughout the second trimester. We have specified normal β2-microglobulin values of each gestational week/period in order to be used in clinical practice. CONCLUSION: We have demonstrated that amnionic fluid β2-microglobulin levels are increased progressively throughout the second trimester.

  3. INSITU BLOTTING - A NOVEL METHOD FOR DIRECT TRANSFER OF NATIVE PROTEINS FROM SECTIONED TISSUE TO BLOTTING MEMBRANE - PROCEDURE AND SOME APPLICATIONS

    NARCIS (Netherlands)

    OKABE, M; NYAKAS, C; BUWALDA, B; LUITEN, PGM

    We describe a novel technique for direct transfer of native proteins from unfixed frozen tissue sections to an immobilizing matrix, e.g., nitrocellulose, polyvinyliden difluoride, or positively charged nylon membranes. Proteins are directly blotted onto the membrane, providing optimal accessibility

  4. Relative Abundance of Integral Plasma Membrane Proteins in Arabidopsis Leaf and Root Tissue Determined by Metabolic Labeling and Mass Spectrometry

    Science.gov (United States)

    Bernfur, Katja; Larsson, Olaf; Larsson, Christer; Gustavsson, Niklas

    2013-01-01

    Metabolic labeling of proteins with a stable isotope (15N) in intact Arabidopsis plants was used for accurate determination by mass spectrometry of differences in protein abundance between plasma membranes isolated from leaves and roots. In total, 703 proteins were identified, of which 188 were predicted to be integral membrane proteins. Major classes were transporters, receptors, proteins involved in membrane trafficking and cell wall-related proteins. Forty-one of the integral proteins, including nine of the 13 isoforms of the PIP (plasma membrane intrinsic protein) aquaporin subfamily, could be identified by peptides unique to these proteins, which made it possible to determine their relative abundance in leaf and root tissue. In addition, peptides shared between isoforms gave information on the proportions of these isoforms. A comparison between our data for protein levels and corresponding data for mRNA levels in the widely used database Genevestigator showed an agreement for only about two thirds of the proteins. By contrast, localization data available in the literature for 21 of the 41 proteins show a much better agreement with our data, in particular data based on immunostaining of proteins and GUS-staining of promoter activity. Thus, although mRNA levels may provide a useful approximation for protein levels, detection and quantification of isoform-specific peptides by proteomics should generate the most reliable data for the proteome. PMID:23990937

  5. Distribution of basic fibroblast growth factor binding sites in various tissue membrane preparations from adult guinea pig

    International Nuclear Information System (INIS)

    Ledoux, D.; Mereau, A.; Dauchel, M.C.; Barritault, D.; Courty, J.

    1989-01-01

    In order to localize a rich source of basic FGF receptor, we examined the distribution of basic FGF binding sites in brain, stomach, lung, spleen, kidney, liver and intestine membrane preparations from adult guinea pig. Comparative binding studies using iodinated basic FGF showed that a specific binding was detected in all the membrane preparations tested. Scatchard plots from iodinated basic FGF competition experiment with native basic FGF in various membrane preparations, suggested the presence of one class of binding sites in some tissues such as liver, kidney, spleen, lung, stomach, and intestine with an apparent dissociation constant (appKD) value ranging from 4 to 7.5 nM and the existence of a second class of higher affinity sites in brain membranes with appKD value of 15 pM. Characterization of these basic FGF high affinity interaction sites was performed using a cross-linking reagent. These results show for the first time that specific interaction sites for basic FGF are widely distributed, suggesting that this growth factor might play a role in the physiological functions of a number of adult organs

  6. Vaginal Lactoferrin Modulates PGE2, MMP-9, MMP-2, and TIMP-1 Amniotic Fluid Concentrations

    Directory of Open Access Journals (Sweden)

    Alessandro Trentini

    2016-01-01

    Full Text Available Inflammation plays an important role in pregnancy, and cytokine and matrix metalloproteases (MMPs imbalance has been associated with premature rupture of membranes and increased risk of preterm delivery. Previous studies have demonstrated that lactoferrin (LF, an iron-binding protein with anti-inflammatory properties, is able to decrease amniotic fluid (AF levels of IL-6. Therefore, we aimed to evaluate the effect of vaginal LF administration on amniotic fluid PGE2 level and MMP-TIMP system in women undergoing genetic amniocentesis. One hundred and eleven women were randomly divided into controls (n=57 or treated with LF 4 hours before amniocentesis (n=54. Amniotic fluid PGE2, active MMP-9 and MMP-2, and TIMP-1 and TIMP-2 concentrations were determined by commercially available assays and the values were normalized by AF creatinine concentration. PGE2, active MMP-9, and its inhibitor TIMP-1 were lower in LF-treated group than in controls (p<0.01, p<0.005, and p<0.001, resp.. Conversely, active MMP-2 (p<0.0001 and MMP-2/TIMP-2 molar ratio (p<0.001 were increased, whilst TIMP-2 was unchanged. Our data suggest that LF administration is able to modulate the inflammatory response following amniocentesis, which may counteract cytokine and prostanoid imbalance that leads to abortion. This trial is registered with Clinical Trial number NCT02695563.

  7. Amniotic fluid protein profiles of intraamniotic inflammatory response to Ureaplasma spp. and other bacteria.

    Directory of Open Access Journals (Sweden)

    Marian Kacerovsky

    Full Text Available OBJECTIVE: This study aimed to evaluate the amniotic fluid protein profiles and the intensity of intraamniotic inflammatory response to Ureaplasma spp. and other bacteria, using the multiplex xMAP technology. METHODS: A retrospective cohort study was undertaken in the Department of Obstetrics and Gynecology, University Hospital Hradec Kralove, Czech Republic. A total of 145 pregnant women with preterm prelabor rupture of membranes between gestational age 24+0 and 36+6 weeks were included in the study. Amniocenteses were performed. The presence of Ureaplasma spp. and other bacteria was evaluated using 16S rRNA gene sequencing. The levels of specific proteins were determined using multiplex xMAP technology. RESULTS: The presence of Ureaplasma spp. and other bacteria in the amniotic fluid was associated with increased levels of interleukin (IL-6, IL-8, IL-10, brain-derived neurotropic factor, granulocyte macrophage colony stimulating factor, monocyte chemotactic protein-1, macrophage inflammatory protein-1, and matrix metalloproteinasis-9. Ureaplasma spp. were also associated with increased levels of neurotropin-3 and triggering receptor expressed on myeloid cells-1. CONCLUSIONS: The presence of Ureaplasma spp. in the amniotic fluid is associated with a slightly different protein profile of inflammatory response, but the intensity of inflammatory response to Ureaplasma spp. is comparable with the inflammatory response to other bacteria.

  8. Amniotic fluid protein profiles of intraamniotic inflammatory response to Ureaplasma spp. and other bacteria.

    Science.gov (United States)

    Kacerovsky, Marian; Celec, Peter; Vlkova, Barbora; Skogstrand, Kristin; Hougaard, David M; Cobo, Teresa; Jacobsson, Bo

    2013-01-01

    This study aimed to evaluate the amniotic fluid protein profiles and the intensity of intraamniotic inflammatory response to Ureaplasma spp. and other bacteria, using the multiplex xMAP technology. A retrospective cohort study was undertaken in the Department of Obstetrics and Gynecology, University Hospital Hradec Kralove, Czech Republic. A total of 145 pregnant women with preterm prelabor rupture of membranes between gestational age 24+0 and 36+6 weeks were included in the study. Amniocenteses were performed. The presence of Ureaplasma spp. and other bacteria was evaluated using 16S rRNA gene sequencing. The levels of specific proteins were determined using multiplex xMAP technology. The presence of Ureaplasma spp. and other bacteria in the amniotic fluid was associated with increased levels of interleukin (IL)-6, IL-8, IL-10, brain-derived neurotropic factor, granulocyte macrophage colony stimulating factor, monocyte chemotactic protein-1, macrophage inflammatory protein-1, and matrix metalloproteinasis-9. Ureaplasma spp. were also associated with increased levels of neurotropin-3 and triggering receptor expressed on myeloid cells-1. The presence of Ureaplasma spp. in the amniotic fluid is associated with a slightly different protein profile of inflammatory response, but the intensity of inflammatory response to Ureaplasma spp. is comparable with the inflammatory response to other bacteria.

  9. Nanofibrous poly(lactide-co-glycolide membranes loaded with diamond nanoparticles as promising substrates for bone tissue engineering

    Directory of Open Access Journals (Sweden)

    Parizek M

    2012-04-01

    Full Text Available Martin Parizek1, Timothy EL Douglas2, Katarina Novotna1, Alexander Kromka3, Mariea A Brady4, Andrea Renzing4, Eske Voss4, Marketa Jarosova3, Lukas Palatinus3, Pavel Tesarek5, Pavla Ryparova5, Vera Lisa1, Ana M dos Santos2, Lucie Bacakova11Department of Biomaterials and Tissue Engineering, Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic; 2Polymer Chemistry and Biomaterials Group, Ghent University, Ghent, Belgium; 3Institute of Physics, Academy of Sciences of the Czech Republic, Prague, Czech Republic; 4Department of Oral and Maxillofacial Surgery, University of Kiel, Kiel, Germany; 5Czech Technical University in Prague, Faculty of Civil Engineering, Prague, Czech RepublicBackground: Nanofibrous scaffolds loaded with bioactive nanoparticles are promising materials for bone tissue engineering.Methods: In this study, composite nanofibrous membranes containing a copolymer of L-lactide and glycolide (PLGA and diamond nanoparticles were fabricated by an electrospinning technique. PLGA was dissolved in a mixture of methylene chloride and dimethyl formamide (2:3 at a concentration of 2.3 wt%, and nanodiamond (ND powder was added at a concentration of 0.7 wt% (about 23 wt% in dry PLGA.Results: In the composite scaffolds, the ND particles were either arranged like beads in the central part of the fibers or formed clusters protruding from the fibers. In the PLGA-ND membranes, the fibers were thicker (diameter 270 ± 9 nm than in pure PLGA meshes (diameter 218 ± 4 nm, but the areas of pores among these fibers were smaller than in pure PLGA samples (0.46 ± 0.02 µm2 versus 1.28 ± 0.09 µm2 in pure PLGA samples. The PLGA-ND membranes showed higher mechanical resistance, as demonstrated by rupture tests of load and deflection of rupture probe at failure. Both types of membranes enabled the attachment, spreading, and subsequent proliferation of human osteoblast-like MG-63 cells to a similar extent, although these

  10. In vitro culture of mouse embryos amniotic fluid ID human

    African Journals Online (AJOL)

    1989-07-15

    Jul 15, 1989 ... Because human amniotic fluid is a physiological, balanced ultrafiltrate, it has been considered as an inexpensive alternative culture medium in. IVF. A study of the development of mouse embryos in human amniotic fluid was undertaken to assess the suitability of this as an optional culture medium in human ...

  11. Assay of Plasma Membrane H+-ATPase in Plant Tissues under Abiotic Stresses.

    Science.gov (United States)

    Janicka, Małgorzata; Wdowikowska, Anna; Kłobus, Grażyna

    2018-01-01

    Plasma membrane (PM) H + -ATPase, which generates the proton gradient across the outer membrane of plant cells, plays a fundamental role in the regulation of many physiological processes fundamental for growth and development of plants. It is involved in the uptake of nutrients from external solutions, their loading into phloem and long-distance transport, stomata aperture and gas exchange, pH homeostasis in cytosol, cell wall loosening, and cell expansion. The crucial role of the enzyme in resistance of plants to abiotic and biotic stress factors has also been well documented. Such great diversity of physiological functions linked to the activity of one enzyme requires a suitable and complex regulation of H + -ATPase. This regulation comprises the transcriptional as well as post-transcriptional levels. Herein, we describe the techniques that can be useful for the analysis of the plasma membrane proton pump modifications at genetic and protein levels under environmental factors.

  12. Plasma membrane proteomic analysis of human Gastric Cancer tissues: revealing flotillin 1 as a marker for Gastric Cancer

    International Nuclear Information System (INIS)

    Gao, Wen; Xu, Jing; Wang, Fuqiang; Zhang, Long; Peng, Rui; Shu, Yongqian; Wu, Jindao; Tang, Qiyun; Zhu, Yunxia

    2015-01-01

    Gastric cancer remains the second leading cause of cancer-related deaths in the world. Successful early gastric cancer detection is hampered by lack of highly sensitive and specific biomarkers. Plasma membrane proteins participate and/or have a central role in the metastatic process of cancer cells and are potentially useful for cancer therapy due to easy accessibility of the targets. In the present research, TMT method followed by mass spectrometry analysis was used to compare the relative expression levels of plasma membrane proteins between noncancer and gastric cancer tissues. Of a total data set that included 501 identified proteins, about 35% of the identified proteins were found to be plasma membrane and associated proteins. Among them, 82 proteins were at least 1.5-fold up- or down-regulated in gastric cancer compared with the adherent normal tissues. A number of markers (e.g. annexin A6, caveolin 1, epidermal growth factor receptor, integrin beta 4) were previously reported as biomarkers of GC. Additionally, several potential biomarkers participated in endocytosis pathway and integrin signaling pathways were firstly identified as differentially expressed proteins in GC samples. Our findings also supported the notion that flotillin 1 is a potential biomarker that could be exploited for molecular imaging-based detection of gastric cancer. Together, the results show that subcellular proteomics of tumor tissue is a feasible and promising avenue for exploring oncogenesis. The online version of this article (doi:10.1186/s12885-015-1343-5) contains supplementary material, which is available to authorized users

  13. A Loss in the Plasma Membrane ATPase Activity and Its Recovery Coincides with Incipient Freeze-Thaw Injury and Postthaw Recovery in Onion Bulb Scale Tissue 1

    Science.gov (United States)

    Arora, Rajeev; Palta, Jiwan P.

    1991-01-01

    Plasma membrane ATPase has been proposed to be functionally altered during early stages of injury caused by a freeze-thaw stress. Complete recovery from freezing injury in onion cells during the postthaw period provided evidence in support of this proposal. During recovery, a simultaneous decrease in ion leakage and disappearance of water soaking (symptoms of freeze-thaw injury) has been noted. Since reabsorption of ions during recovery must be an active process, recovery of plasma membrane ATPase (active transport system) functions has been implicated. In the present study, onion (Allium cepa L. cv Downing Yellow Globe) bulbs were subjected to a freeze-thaw stress which resulted in a reversible (recoverable) injury. Plasma membrane ATPase activity in the microsomes (isolated from the bulb scales) and ion leakage rate (efflux/hour) from the same scale tissue were measured immediately following thawing and after complete recovery. In injured tissue (30-40% water soaking), plasma membrane ATPase activity was reduced by about 30% and this was paralleled by about 25% higher ion leakage rate. As water soaking disappeared during recovery, the plasma membrane ATPase activity and the ion leakage rate returned to about the same level as the respective controls. Treatment of freeze-thaw injured tissue with vanadate, a specific inhibitor of plasma membrane ATPase, during postthaw prevented the recovery process. These results indicate that recovery of freeze-injured tissue depends on the functional activity of plasma membrane ATPase. PMID:16668063

  14. Polymeric hollow fiber membranes for bioartificial organs and tissue engineering applications

    NARCIS (Netherlands)

    Diban-Ibrahim Gomez, Nazely; Stamatialis, Dimitrios

    2014-01-01

    Polymeric hollow fiber (HF) membranes are commercially available, i.e. microfiltration and ultrafiltration cartridges or reverse osmosis and gas separation modules, to be applied for separation purposes in industry, for instance to recover valuable raw materials or products, or for the treatment of

  15. The adult brain tissue response to hollow fiber membranes of varying surface architecture with or without cotransplanted cells

    Science.gov (United States)

    Zhang, Ning

    A variety of biomaterials have been chronically implanted into the central nervous system (CNS) for repair or therapeutic purposes. Regardless of the application, chronic implantation of materials into the CNS induces injury and elicits a wound healing response, eventually leading to the formation of a dense extracellular matrix (ECM)-rich scar tissue that is associated with the segregation of implanted materials from the surrounding normal tissue. Often this reaction results in impaired performance of indwelling CNS devices. In order to enhance the performance of biomaterial-based implantable devices in the CNS, this thesis investigated whether adult brain tissue response to implanted biomaterials could be manipulated by changing biomaterial surface properties or further by utilizing the biology of co-transplanted cells. Specifically, the adult rat brain tissue response to chronically implanted poly(acrylonitrile-vinylchloride) (PAN-PVC) hollow fiber membranes (HFMs) of varying surface architecture were examined temporally at 2, 4, and 12 weeks postimplantation. Significant differences were discovered in the brain tissue response to the PAN-PVC HFMs of varying surface architecture at 4 and 12 weeks. To extend this work, whether the soluble factors derived from a co-transplanted cellular component further affect the brain tissue response to an implanted HFM in a significant way was critically exploited. The cells used were astrocytes, whose ability to influence scar formation process following CNS injury by physical contact with the host tissue had been documented in the literature. Data indicated for the first time that astrocyte-derived soluble factors ameliorate the adult brain tissue reactivity toward HFM implants in an age-dependent manner. While immature astrocytes secreted soluble factors that suppressed the brain tissue reactivity around the implants, mature astrocytes secreted factors that enhanced the gliotic response. These findings prove the feasibility

  16. The evolution of tail weaponization in amniotes.

    Science.gov (United States)

    Arbour, Victoria M; Zanno, Lindsay E

    2018-01-31

    Weaponry, for the purpose of intraspecific combat or predator defence, is one of the most widespread animal adaptations, yet the selective pressures and constraints governing its phenotypic diversity and skeletal regionalization are not well understood. Here, we investigate the evolution of tail weaponry in amniotes, a rare form of weaponry that nonetheless evolved independently among a broad spectrum of life including mammals, turtles and dinosaurs. Using phylogenetic comparative methods, we test for links between morphology, ecology and behaviour in extant amniotes known to use the tail as a weapon, and in extinct taxa bearing osseous tail armaments. We find robust ecological and morphological correlates of both tail lashing behaviour and bony tail weaponry, including large body size, body armour and herbivory, suggesting these life-history parameters factor into the evolution of antipredator behaviours and tail armaments. We suggest that the evolution of tail weaponry is rare because large, armoured herbivores are uncommon in extant terrestrial faunas, as they have been throughout evolutionary history. © 2018 The Author(s).

  17. Rapid Preparation of a Plasma Membrane Fraction: Western Blot Detection of Translocated Glucose Transporter 4 from Plasma Membrane of Muscle and Adipose Cells and Tissues.

    Science.gov (United States)

    Yamamoto, Norio; Yamashita, Yoko; Yoshioka, Yasukiyo; Nishiumi, Shin; Ashida, Hitoshi

    2016-08-01

    Membrane proteins account for 70% to 80% of all pharmaceutical targets, indicating their clinical relevance and underscoring the importance of identifying differentially expressed membrane proteins that reflect distinct disease properties. The translocation of proteins from the bulk of the cytosol to the plasma membrane is a critical step in the transfer of information from membrane-embedded receptors or transporters to the cell interior. To understand how membrane proteins work, it is important to separate the membrane fraction of cells. This unit provides a protocol for rapidly obtaining plasma membrane fractions for western blot analysis. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

  18. Isolation, Characterization, Cryopreservation of Human Amniotic Stem Cells and Differentiation to Osteogenic and Adipogenic Cells.

    Directory of Open Access Journals (Sweden)

    Shiva Gholizadeh-Ghaleh Aziz

    Full Text Available Human stem cells and progenitor cells can be used to treat cancer and replace dysfunctional cells within a tissue or organ. The objective of this study was to identify the appropriate cells type in regenerative medicine and targeted therapy. As an alternative to embryonic and bone marrow stem cells, we examined human amniotic fluid stem cells (hAFSCs, one of the potential source of multipotent stem cells isolated from both cell pellet (using single-stage method, and supernatant of human amniotic fluid. Source of isolation and unique property of the cells emphasize that these cells are one of the promising new tools in therapeutic field. Double sources for isolation and availability of the left over samples in diagnostic laboratory at the same time have less legal and ethical concerns compared with embryonic stem cell studies. Cells were isolated, cultured for 18th passage for 6 months and characterized using qPCR and flow cytometry. Cells showed good proliferative ability in culture condition. The cells successfully differentiated into the adipogenic and osteogenic lineages. Based on these findings, amniotic fluid can be considered as an appropriate and convenient source of human amniotic fluid stem cells. These cells provide potential tools for therapeutic applications in the field of regenerative medicine. To get a better understanding of crosstalk between Oct4/NANOG with osteogenesis and adipogenesis, we used network analysis based on Common Targets algorithm and Common Regulators algorithm as well as subnetwork discovery based on gene set enrichment. Network analysis highlighted the possible role of MIR 302A and MIR let-7g. We demonstrated the high expression of MIR 302A and low expression of MIR let7g in hAFSCs by qPCR.

  19. [Clinical features and expression of PLA(2)R in renal tissue with idiopathic membranous nephropathy in children].

    Science.gov (United States)

    Dong, Y F; Sun, L W; Zhang, B; Kuang, X Y; Niu, X L; Kang, Y L; Hao, S; Wang, P; Li, Z; Zhu, G H; Huang, W Y; Wu, Y

    2018-03-02

    Objective: To explore the clinical features and expression of PLA(2)R in renal tissue of children with idiopathic membranous nephropathy. Methods: Retrospective study was performed in patients with membranous nephropathy diagnosed through renal biopsy and the follow-up time was at least half a year in Shanghai Children ' s Hospital from January 2010 to February 2017. We compared their clinicopathological and pathological findings of IMN. Indirect immunofluorescence assay was used to detect glomerular PLA(2)R expression. We analyzed the differences of clinical features between the PLA(2)R negative and positive groups. T test, rank-sum test and Fisher exact test were used. Results: Eleven cases had hematuria and proteinuria, 9 cases presented with nephrotic syndrome, and 2 cases showed isolated proteinuria. Of the 22 cases of children with IMN, 16 patients had complete remission (complete remission rate was 72.8%), and 22 patients had partial remission. The renal function of all cases was normal and in all cases the estimated glomerular filtration rate was > 90 ml/(min·1.73m(2)). Of 22 cases with IMN, 7 cases were PLA(2)R-positive in renal tissue and 15 cases were PLA(2)R-negative. The age of positive group (10 years old) was older than the negative group (6 years old)( Z= -2.483, PPLA(2)R in renal tissue of children with IMN was about 32%. The age of PLA(2)R positive group was older than the negative group. And the time of urine protein turning to negative in positive group was longer than that in the negative group.

  20. Amniotic fluid embolism and isolated coagulopathy: atypical presentation of amniotic fluid embolism.

    LENUS (Irish Health Repository)

    Awad, I T

    2012-02-03

    A 41-year-old multigravida presented at 32 weeks of gestation with polyhydramnios and an anencephalic fetus. Abnormal bleeding as a result of disseminated intravascular coagulation complicated an emergency Caesarean section for severe abdominal pain thought to be due to uterine rupture. Massive transfusion with blood products was necessary and the abdomen packed to control bleeding. The patient was transferred to the intensive care unit where she made a slow but complete recovery. Amniotic fluid embolism with atypical presentation of isolated coagulopathy is the likely diagnosis in this case. The case serves to demonstrate that amniotic fluid embolism may present with symptoms and signs other than the classical pattern of dyspnoea, cyanosis and hypotension.

  1. Oral Rehabilitation for Amniotic Band Syndrome: An Unusual Presentation.

    Science.gov (United States)

    Hotwani, Kavita; Sharma, Krishna

    2015-01-01

    Amniotic band syndrome (ABS) is a congenital disorder caused by entrapment of fetal parts in fibrous amniotic bands while in utero. The syndrome is underdiagnosed and its presentation is variable. The syndrome has been well described in the pediatric, orthopedic and obstetric literature; however, despite the discernable craniomaxillofacial involvement, ABS has not been reported in the dental literature very often. The present report describes a case of a patient with ABS and concomitant dental findings. How to cite this article: Hotwani K, Sharma K. Oral Rehabilitation for Amniotic Band Syndrome: An Unusual Presentation. Int J Clin Pediatr Dent 2015;8(1):55-57.

  2. Layer-by-layer bioassembly of cellularized polylactic acid porous membranes for bone tissue engineering

    NARCIS (Netherlands)

    Guduric, Vera; Metz, Carole; Siadous, Robin; Bareille, Reine; Levato, Riccardo; Engel, Elisabeth; Fricain, Jean-Christophe; Devillard, Raphaël; Luzanin, Ognjan; Catros, Sylvain

    2017-01-01

    The conventional tissue engineering is based on seeding of macroporous scaffold on its surface ("top-down" approach). The main limitation is poor cell viability in the middle of the scaffold due to poor diffusion of oxygen and nutrients and insufficient vascularization. Layer-by-Layer (LBL)

  3. High MMP-9 and TNF-α expression increase in preterm premature rupture of membranes

    Directory of Open Access Journals (Sweden)

    Sri Sulistyowati

    2016-05-01

    Expression of MMP-9 and TNF-α was higher in the amniotic membrane of preterm delivery subjects with PROM than in preterm delivery subjects without PROM and can thus be used as predictor to avoid PPROM.

  4. GROUP B STREPTOCOCCUS CIRCUMVENTS NEUTROPHILS AND NEUTROPHIL EXTRACELLULAR TRAPS DURING AMNIOTIC CAVITY INVASION AND PRETERM LABOR

    Science.gov (United States)

    Boldenow, Erica; Gendrin, Claire; Ngo, Lisa; Bierle, Craig; Vornhagen, Jay; Coleman, Michelle; Merillat, Sean; Armistead, Blair; Whidbey, Christopher; Alishetti, Varchita; Santana-Ufret, Veronica; Ogle, Jason; Gough, Michael; Srinouanprachanh, Sengkeo; MacDonald, James W; Bammler, Theo K; Bansal, Aasthaa; Liggitt, H. Denny; Rajagopal, Lakshmi; Waldorf, Kristina M Adams

    2016-01-01

    Preterm birth is a leading cause of neonatal morbidity and mortality. Although microbial invasion of the amniotic cavity (MIAC) is associated with the majority of early preterm births, the temporal events that occur during MIAC and preterm labor are not known. Group B Streptococci (GBS) are β-hemolytic, gram-positive bacteria, which commonly colonize the vagina but have been recovered from the amniotic fluid in preterm birth cases. To understand temporal events that occur during MIAC, we utilized a unique chronically catheterized nonhuman primate model that closely emulates human pregnancy. This model allows monitoring of uterine contractions, timing of MIAC and immune responses during pregnancy-associated infections. Here, we show that adverse outcomes such as preterm labor, MIAC, and fetal sepsis were observed more frequently during infection with hemolytic GBS when compared to nonhemolytic GBS. Although MIAC was associated with systematic progression in chorioamnionitis beginning with chorionic vasculitis and progressing to neutrophilic infiltration, the ability of the GBS hemolytic pigment toxin to induce neutrophil cell death and subvert killing by neutrophil extracellular traps (NETs) in placental membranes in vivo facilitated MIAC and fetal injury. Furthermore, compared to maternal neutrophils, fetal neutrophils exhibit decreased neutrophil elastase activity and impaired phagocytic functions to GBS. Collectively, our studies demonstrate how a unique bacterial hemolytic lipid toxin enables GBS to circumvent neutrophils and NETs in placental membranes to induce fetal injury and preterm labor. PMID:27819066

  5. Progress in the processing of radioesterilized tissue

    International Nuclear Information System (INIS)

    Zarate S, H; Espinoza B, J; Ribbeck N, J; Vargas Q, M; Gutierrez D, K

    2003-01-01

    Since 1996, the Chilean Nuclear Energy Commission has been carrying out work to implement the first Radiosterilized Tissue Processing Laboratory (RTPL) in Chile, in order to introduce the use of sterilized biological tissue for clinical application. The International Atomic Energy Agency (IAEA) has provided collaboration and technical assistance for this work. The processing of biological tissues has been done in conjunction with physicians from different state hospital centers, mostly in the Metropolitan Region. Among the tissues primarily processed are allografts such as frozen human skin at - 80 o C, freeze-dried human bone and amniotic membrane. We have also been working with xenograft developments such as freeze-dried pig skin and demineralized ground cow bone. All these tissues are sterilized by means of gamma radiation, in order to obtain a sterility assurance level (SAL) of 10 -6 . This laboratory has already completed various stages, from the beginning when it was only just an idea up to the production stage where a large quantity of processed tissues have been delivered to physicians of different specialties, resulting in a contribution to medical science as well as to the treatment quality of a great many patients. The preliminary results and the opinions of those physicians who have used the processed products from our laboratory have encouraged us to continue developing new products, thus enlarging the scope of application (author)

  6. Membrane Lipid Replacement for chronic illnesses, aging and cancer using oral glycerolphospholipid formulations with fructooligosaccharides to restore phospholipid function in cellular membranes, organelles, cells and tissues.

    Science.gov (United States)

    Nicolson, Garth L; Ash, Michael E

    2017-09-01

    Membrane Lipid Replacement is the use of functional, oral supplements containing mixtures of cell membrane glycerolphospholipids, plus fructooligosaccharides (for protection against oxidative, bile acid and enzymatic damage) and antioxidants, in order to safely replace damaged, oxidized, membrane phospholipids and restore membrane, organelle, cellular and organ function. Defects in cellular and intracellular membranes are characteristic of all chronic medical conditions, including cancer, and normal processes, such as aging. Once the replacement glycerolphospholipids have been ingested, dispersed, complexed and transported, while being protected by fructooligosaccharides and several natural mechanisms, they can be inserted into cell membranes, lipoproteins, lipid globules, lipid droplets, liposomes and other carriers. They are conveyed by the lymphatics and blood circulation to cellular sites where they are endocytosed or incorporated into or transported by cell membranes. Inside cells the glycerolphospholipids can be transferred to various intracellular membranes by lipid globules, liposomes, membrane-membrane contact or by lipid carrier transfer. Eventually they arrive at their membrane destinations due to 'bulk flow' principles, and there they can stimulate the natural removal and replacement of damaged membrane lipids while undergoing further enzymatic alterations. Clinical trials have shown the benefits of Membrane Lipid Replacement in restoring mitochondrial function and reducing fatigue in aged subjects and chronically ill patients. Recently Membrane Lipid Replacement has been used to reduce pain and other symptoms as well as removing hydrophobic chemical contaminants, suggesting that there are additional new uses for this safe, natural medicine supplement. This article is part of a Special Issue entitled: Membrane Lipid Therapy: Drugs Targeting Biomembranes edited by Pablo V. Escribá. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights

  7. Evaluation of the tissue toxicity of antiseptics by the hen's egg test on the chorioallantoic membrane (HETCAM)

    Science.gov (United States)

    2010-01-01

    Background Antiseptics are frequently used for the prophylaxis and treatment of local infections of chronic wounds. Whereas local antiseptics in general have a positive effect on wound healing an uncritical use may impair wound healing due to toxic side effects. Objective We sought to assess the vascular irritation potential of different antiseptic solutions and ointments commonly used for short and long term application as a measure of tissue toxicity. Method The vascular irritation was evaluated by the hen's egg test (HET) on the chorioallantoic membrane (CAM). The effects on the vessels of a mucous membrane were directly assessed by stereomicroscopic observation in vivo. Results Severe CAM irritation was observed after short-term applications of 1% octenidin-2HCl (Octeni sept™), 72% isopropanol (Cutasept™), 0.35% chloroxylenol (Dettol™) and 10% PVP-I ointment (Betaisodona™). Medium irritations were observed for 10% PVP-I solution (Betaisodona™), 3% lysosomal PVP-I ointment (Repithel™), 1.8% cadexomer-iodine ointment (Iodosorb™) and 1% cadexomer-iodine pellets (Iodosorb™). Finally, slight irritations were observed for 1% PVP-I solution (Betaisodona™), 0.1% polyhexanid plus betain (Prontosan™) and 1% silver-sulfadiazine ointment (Flammazine™), whereas 0.04% polyhexanid solution (Lavanid™), washings from sterile maggots of Lucilia sericata and filtrated enzymes from Clostridium histolyticum (Iruxol-N™) showed no effects of irritation. In the long-term approaches, no vascular irritations were found for polyhexanid, washings from Lucilia sericata and enzyme filtrations from Clostridium histolyticum. Conclusion The vascular injuries caused by the studied antiseptics are an indirect indicator of their tissue toxicity. Strikingly, even therapeutic substances, which have been regarded as safe in their application for the treatment of chronic wounds in clinical studies, showed severe irritations on the CAM. We suggest that agents with no or low

  8. Evaluation of the tissue toxicity of antiseptics by the hen's egg test on the chorioallantoic membrane (HETCAM

    Directory of Open Access Journals (Sweden)

    Marquardt C

    2010-05-01

    Full Text Available Abstract Background Antiseptics are frequently used for the prophylaxis and treatment of local infections of chronic wounds. Whereas local antiseptics in general have a positive effect on wound healing an uncritical use may impair wound healing due to toxic side effects. Objective We sought to assess the vascular irritation potential of different antiseptic solutions and ointments commonly used for short and long term application as a measure of tissue toxicity. Method The vascular irritation was evaluated by the hen's egg test (HET on the chorioallantoic membrane (CAM. The effects on the vessels of a mucous membrane were directly assessed by stereomicroscopic observation in vivo. Results Severe CAM irritation was observed after short-term applications of 1% octenidin-2HCl (Octeni sept™, 72% isopropanol (Cutasept™, 0.35% chloroxylenol (Dettol™ and 10% PVP-I ointment (Betaisodona™. Medium irritations were observed for 10% PVP-I solution (Betaisodona™, 3% lysosomal PVP-I ointment (Repithel™, 1.8% cadexomer-iodine ointment (Iodosorb™ and 1% cadexomer-iodine pellets (Iodosorb™. Finally, slight irritations were observed for 1% PVP-I solution (Betaisodona™, 0.1% polyhexanid plus betain (Prontosan™ and 1% silver-sulfadiazine ointment (Flammazine™, whereas 0.04% polyhexanid solution (Lavanid™, washings from sterile maggots of Lucilia sericata and filtrated enzymes from Clostridium histolyticum (Iruxol-N™ showed no effects of irritation. In the long-term approaches, no vascular irritations were found for polyhexanid, washings from Lucilia sericata and enzyme filtrations from Clostridium histolyticum. Conclusion The vascular injuries caused by the studied antiseptics are an indirect indicator of their tissue toxicity. Strikingly, even therapeutic substances, which have been regarded as safe in their application for the treatment of chronic wounds in clinical studies, showed severe irritations on the CAM. We suggest that agents

  9. Testosterone determination in amniotic fluid for sec diagnosis

    International Nuclear Information System (INIS)

    Quiroa C, M.M.

    1985-11-01

    This study was carried on 50 samples of amniotic fluid obtained from pregnant patients with gestation old of 38 to 40 weeks; diagnosis of the foetus sex was made by measuring the testosterone levels by radioimmunoassay technique. It was found that 94% of the cases were correctly diagnosed. The testosterone levels found in the amniotic fluid of male and female foetus were significantly different (L<0.01) these confirm the efficacy of the method. (author)

  10. Pup retrieval and maternal attraction to canine amniotic fluids.

    Science.gov (United States)

    Dunbar, I; Ranson, E; Buehler, M

    1981-10-01

    Three purebred female beagles were observed with both their first and second litters. Dams were given three separate simultaneous-choice retrieval tests: In addition, the maternal response to amniotic fluids was observed when one pup in the litter was treated with amniotic fluids and all other pups were treated with water. There was no evidence to suggest that pups were retrieved preferentially on the basis of their sex. One female retrieved her own but not alien pups, whereas another female made no such discrimination and readily retrieved alien pups in addition to her own. Retrieval behavior developed at the time of each whelping and normally lasted until the pups were about 5 days old, although a bitch would retrieve younger (alien) pups up to 14 days post partum. Maternal bitches were strongly attracted towards amniotic fluids: they investigated pups daubed with amniotic fluids to a significantly greater extent than control pups treated with water. The attraction of maternal females towards amniotic fluids developed at the time of each whelping and persisted for up to 30 days, well beyond the time that a whelping bitch would normally be exposed to her own fetal fluids. A possible role for amniotic fluids in the development of maternal behaviour and the establishment of the maternal/puppy bond is discussed. Copyright © 1981. Published by Elsevier B.V.

  11. Detergent resistant membrane-associated IDE in brain tissue and cultured cells: Relevance to Aβ and insulin degradation

    Directory of Open Access Journals (Sweden)

    Castaño Eduardo M

    2008-12-01

    Full Text Available Abstract Background Insulin degrading enzyme (IDE is implicated in the regulation of amyloid β (Aβ steady-state levels in the brain, and its deficient expression and/or activity may be a risk factor in sporadic Alzheimer's disease (AD. Although IDE sub-cellular localization has been well studied, the compartments relevant to Aβ degradation remain to be determined. Results Our results of live immunofluorescence, immuno gold electron-microscopy and gradient fractionation concurred to the demonstration that endogenous IDE from brain tissues and cell cultures is, in addition to its other localizations, a detergent-resistant membrane (DRM-associated metallopeptidase. Our pulse chase experiments were in accordance with the existence of two pools of IDE: the cytosolic one with a longer half-life and the membrane-IDE with a faster turn-over. DRMs-associated IDE co-localized with Aβ and its distribution (DRMs vs. non-DRMs and activity was sensitive to manipulation of lipid composition in vitro and in vivo. When IDE was mis-located from DRMs by treating cells with methyl-β-cyclodextrin (MβCD, endogenous Aβ accumulated in the extracellular space and exogenous Aβ proteolysis was impaired. We detected a reduced amount of IDE in DRMs of membranes isolated from mice brain with endogenous reduced levels of cholesterol (Chol due to targeted deletion of one seladin-1 allele. We confirmed that a moderate shift of IDE from DRMs induced a substantial decrement on IDE-mediated insulin and Aβ degradation in vitro. Conclusion Our results support the notion that optimal substrate degradation by IDE may require its association with organized-DRMs. Alternatively, DRMs but not other plasma membrane regions, may act as platforms where Aβ accumulates, due to its hydrophobic properties, reaching local concentration close to its Km for IDE facilitating its clearance. Structural integrity of DRMs may also be required to tightly retain insulin receptor and IDE for

  12. Photodynamic damage to cartilage and synovial tissue grafted on a chick's chorioallantoic membrane

    Science.gov (United States)

    Fisher, M.; Nahir, A. M.; Kimel, Sol

    1997-09-01

    Rheumatoid arthritis (RA) is a chronic inflammatory disease of the synovial joints causing pain deformities and disability. The highly vascular inflamed synovium has aggressive and destructive characteristics, it invades, erodes and gradually destroys cartilage and underlying bone. Photodynamic therapy (PDT) was performed using the chick chorioallantoic membrane (CAM) model to investigate the vitality of synovium and cartilage implanted on the CAM. Synovium, obtained from human patients, was grafted onto the CAM; gross microscopy and histology proved its vitality 7 days post grafting. Cartilage obtained from rabbit knee joint was also maintained on the CAM for 7 days. Its vitality was demonstrated by histology and by measuring metabolic and enzymatic activity of cartilage cells (chondrocytes) as well as the collagen and proteoglycans content. Selective PDT was performed using aluminum phthalocyanine tetrasulfonate (AlPcS4), a hydrophilic compound, soluble in biological solutions, as a photosensitizer. After irradiation with a diode laser (lambda equals 670 nm, 10 mW) damage was observed in vascularized synovium grafts, whereas avascular cartilage remained intact.

  13. Maternal Azithromycin Therapy for Ureaplasma Intra-Amniotic Infection Delays Preterm Delivery and Reduces Fetal Lung Injury in a Primate Model

    Science.gov (United States)

    Grigsby, Peta L.; Novy, Miles J.; Sadowsky, Drew W.; Morgan, Terry K.; Long, Mary; Acosta, Ed; Duffy, Lynn B; Waites, Ken B.

    2012-01-01

    Objective We assessed the efficacy of a maternal multi–dose azithromycin (AZI) regimen, with and without anti–inflammatory agents to delay preterm birth and to mitigate fetal lung injury associated with Ureaplasma parvum intra–amniotic infection (IAI). Study Design Long–term catheterized rhesus monkeys (n=16) received intra–amniotic inoculation of U. parvum (107 CFU/ml, serovar 1). After contraction onset, rhesus monkeys received either no treatment (n=6); AZI (12.5mg/kg, q12h, IV for 10 days; n=5); or AZI plus dexamethasone (DEX) and indomethacin (INDO; n=5). Outcomes included amniotic fluid pro–inflammatory mediators, U. parvum cultures & PCR, AZI pharmacokinetics and the extent of fetal lung inflammation. Results Maternal AZI therapy eradicated U. parvum IAI from the amniotic fluid within 4 days. Placenta and fetal tissues were 90% culture negative at delivery. AZI therapy significantly delayed preterm delivery and prevented advanced fetal lung injury, although residual acute chorioamnionitis persisted. Conclusions Specific maternal antibiotic therapy can eradicate U. parvum from the amniotic fluid and key fetal organs, with subsequent prolongation of pregnancy which provides a therapeutic window of opportunity to effectively reduce the severity of fetal lung injury. PMID:23111115

  14. Recruitment of host's progenitor cells to sites of human amniotic fluid stem cells implantation.

    Science.gov (United States)

    Mirabella, Teodelinda; Poggi, Alessandro; Scaranari, Monica; Mogni, Massimo; Lituania, Mario; Baldo, Chiara; Cancedda, Ranieri; Gentili, Chiara

    2011-06-01

    The amniotic fluid is a new source of multipotent stem cells with a therapeutic potential for human diseases. Cultured at low cell density, human amniotic fluid stem cells (hAFSCs) were still able to generate colony-forming unit-fibroblast (CFU-F) after 60 doublings, thus confirming their staminal nature. Moreover, after extensive in vitro cell expansion hAFSCs maintained a stable karyotype. The expression of genes, such as SSEA-4, SOX2 and OCT3/4 was confirmed at early and later culture stage. Also, hAFSCs showed bright expression of mesenchymal lineage markers and immunoregulatory properties. hAFSCs, seeded onto hydroxyapatite scaffolds and subcutaneously implanted in nude mice, played a pivotal role in mounting a response resulting in the recruitment of host's progenitor cells forming tissues of mesodermal origin such as fat, muscle, fibrous tissue and immature bone. Implanted hAFSCs migrated from the scaffold to the skin overlying implant site but not to other organs. Given their in vivo: (i) recruitment of host progenitor cells, (ii) homing towards injured sites and (iii) multipotentiality in tissue repair, hAFSCs are a very appealing reserve of stem cells potentially useful for clinical application in regenerative medicine. Copyright © 2011 Elsevier Ltd. All rights reserved.

  15. [Influence of delta-sleep inducing peptide on the state of lysosomal membranes and intensity of lysosomal proteolysis in different rat tissues during physiological aging of the organism].

    Science.gov (United States)

    Kutilin, D S; Bondarenko, T I; Mikhaleva, I I

    2014-01-01

    It is shown that subcutaneous injection of exogenous delta-sleep inducing peptide (DSIP) to rats aged 2-24 months in a dose of 100 μg/kg animal body weight by courses of 5 consecutive days per month has a stabilizing effect on the state of lysosomal membranes in rat tissues (brain, heart muscle and liver) at different ontogenetic stages, and this effect is accompanied by increasing intensity of lysosomal proteolysis in these tissues.

  16. Application of human amniotic mesenchymal cells as an allogeneic transplantation cell source in bone regenerative therapy

    International Nuclear Information System (INIS)

    Tsuno, Hiroaki; Yoshida, Toshiko; Nogami, Makiko; Koike, Chika; Okabe, Motonori; Noto, Zenko; Arai, Naoya; Noguchi, Makoto; Nikaido, Toshio

    2012-01-01

    Autogenous mesenchymal stem cells (MSCs) have therapeutic applications in bone regenerative therapy due to their pluripotency. However, the ability of MSCs to proliferate and differentiate varies between donors. Furthermore, alternative sources of MSCs are required for patients with contraindications to autogenous cell therapy. The aim of this study was to evaluate the potential of mesenchymal cells from the human amniotic membrane (HAM) as a source of cells for allogeneic transplantation in bone regenerative therapy. Cells that retained a proliferative capacity of more than 50 population doubling level were distinguished from other HAM cells as HAMα cells and induced to osteogenic status—their in vivo osteogenesis was subsequently investigated in rats. It was found that HAMα cells were spindle shaped and were positive for MSC markers and negative for hematopoietic stem cell markers. Alkaline phosphatase activity and calcium deposition increased with osteogenic status of HAMα cells. The expression of osteocalcin mRNA was increased in HAMα cells cultured on calcium phosphate scaffolds. Moreover, xenografted HAMα cells remained viable and produced extracellular matrix for several weeks. Thus, this study suggests that human amniotic mesenchymal cells possess osteogenic differentiation potential and could be applied to allogeneic transplantation in bone regenerative therapy. - Highlights: ► Human amniotic mesenchymal cells include cells (HAMα cells) that have the properties of MSCs. ► HAMα cells have excellent osteogenic differentiation potential. ► Osteogenic differentiation ability of HAMα was amplified by calcium phosphate scaffolds. ► HAMα cells can be applicable to allogeneic cell transplantation in bone regenerative therapy.

  17. Application of human amniotic mesenchymal cells as an allogeneic transplantation cell source in bone regenerative therapy

    Energy Technology Data Exchange (ETDEWEB)

    Tsuno, Hiroaki [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Yoshida, Toshiko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Nogami, Makiko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Orthopedic Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Koike, Chika; Okabe, Motonori [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Noto, Zenko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Arai, Naoya; Noguchi, Makoto [Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Nikaido, Toshio, E-mail: tnikaido@med.u-toyama.ac.jp [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan)

    2012-12-01

    Autogenous mesenchymal stem cells (MSCs) have therapeutic applications in bone regenerative therapy due to their pluripotency. However, the ability of MSCs to proliferate and differentiate varies between donors. Furthermore, alternative sources of MSCs are required for patients with contraindications to autogenous cell therapy. The aim of this study was to evaluate the potential of mesenchymal cells from the human amniotic membrane (HAM) as a source of cells for allogeneic transplantation in bone regenerative therapy. Cells that retained a proliferative capacity of more than 50 population doubling level were distinguished from other HAM cells as HAM{alpha} cells and induced to osteogenic status-their in vivo osteogenesis was subsequently investigated in rats. It was found that HAM{alpha} cells were spindle shaped and were positive for MSC markers and negative for hematopoietic stem cell markers. Alkaline phosphatase activity and calcium deposition increased with osteogenic status of HAM{alpha} cells. The expression of osteocalcin mRNA was increased in HAM{alpha} cells cultured on calcium phosphate scaffolds. Moreover, xenografted HAM{alpha} cells remained viable and produced extracellular matrix for several weeks. Thus, this study suggests that human amniotic mesenchymal cells possess osteogenic differentiation potential and could be applied to allogeneic transplantation in bone regenerative therapy. - Highlights: Black-Right-Pointing-Pointer Human amniotic mesenchymal cells include cells (HAM{alpha} cells) that have the properties of MSCs. Black-Right-Pointing-Pointer HAM{alpha} cells have excellent osteogenic differentiation potential. Black-Right-Pointing-Pointer Osteogenic differentiation ability of HAM{alpha} was amplified by calcium phosphate scaffolds. Black-Right-Pointing-Pointer HAM{alpha} cells can be applicable to allogeneic cell transplantation in bone regenerative therapy.

  18. Toward guided tissue and bone regeneration: morphology, attachment, proliferation, and migration of cells cultured on collagen barrier membranes. A systematic review.

    NARCIS (Netherlands)

    Behring, J.; Junker, R.; Walboomers, X.F.; Chessnut, B.; Jansen, J.A.

    2008-01-01

    Collagen barrier membranes are frequently used in both guided tissue regeneration (GTR) and guided bone regeneration (GBR). Collagen used for these devices is available from different species and is often processed to alter the properties of the final product. This is necessary because unprocessed

  19. Micro pore arrays in free standing cyclic olefin copolymer membranes: fabrication and surface functionalization strategies for in-vitro barrier tissue models

    Science.gov (United States)

    Gel, M.; Kandasamy, S.; Cartledge, K.; Be, C. L.; Haylock, D.

    2013-12-01

    In recent years there has been growing interest in micro engineered in-vitro models of tissues and organs. These models are designed to mimic the in-vivo like physiological conditions with a goal to study human physiology in an organ-specific context or to develop in-vitro disease models. One of the challenges in the development of these models is the formation of barrier tissues in which the permeability is controlled locally by the tissues cultured at the interface. In-vitro models of barrier tissues are typically created by generating a monolayer of cells grown on thin porous membranes. This paper reports a robust preparation method for free standing porous cyclic olefin copolymer (COC) membranes. We also demonstrate that gelatin coated membranes facilitate formation of highly confluent monolayer of HUVECs. Membranes with thickness in the range of 2-3 um incorporating micro pores with diameter approximately 20 um were fabricated and integrated with microfluidic channels. The performance of the device was demonstrated with a model system mimicking the endothelial barrier in bone marrow sinusoids.

  20. Tissue

    Directory of Open Access Journals (Sweden)

    David Morrissey

    2012-01-01

    Full Text Available Purpose. In vivo gene therapy directed at tissues of mesenchymal origin could potentially augment healing. We aimed to assess the duration and magnitude of transene expression in vivo in mice and ex vivo in human tissues. Methods. Using bioluminescence imaging, plasmid and adenoviral vector-based transgene expression in murine quadriceps in vivo was examined. Temporal control was assessed using a doxycycline-inducible system. An ex vivo model was developed and optimised using murine tissue, and applied in ex vivo human tissue. Results. In vivo plasmid-based transgene expression did not silence in murine muscle, unlike in liver. Although maximum luciferase expression was higher in muscle with adenoviral delivery compared with plasmid, expression reduced over time. The inducible promoter cassette successfully regulated gene expression with maximum levels a factor of 11 greater than baseline. Expression was re-induced to a similar level on a temporal basis. Luciferase expression was readily detected ex vivo in human muscle and tendon. Conclusions. Plasmid constructs resulted in long-term in vivo gene expression in skeletal muscle, in a controllable fashion utilising an inducible promoter in combination with oral agents. Successful plasmid gene transfection in human ex vivo mesenchymal tissue was demonstrated for the first time.

  1. Intra-uterine tissue engineering of full-thickness skin defects in a fetal sheep model

    NARCIS (Netherlands)

    Hosper, Nynke A.; Eggink, Alex J.; Roelofs, Luc A. J.; Wijnen, Rene M. H.; van Luyn, Marja J. A.; Bank, Ruud A.; Harmsen, Martin C.; Geutjes, Paul J.; Daamen, Willeke F.; van Kuppevelt, Toin H.; Tiemessen, Dorien M.; Oosterwijk, Egbert; Crevels, Jane J.; Blokx, Willeke A. M.; Lotgering, Fred K.; van den Berg, Paul P.; Feitz, Wout F. J.

    In spina bifida the neural tube fails to close during the embryonic period and it is thought that prolonged exposure of the unprotected spinal cord to the amniotic fluid during pregnancy causes additional neural damage. Intra-uterine repair might protect the neural tissue from exposure to amniotic

  2. The Role of the Multiple Banded Antigen of Ureaplasma parvum in Intra-Amniotic Infection: Major Virulence Factor or Decoy?

    Science.gov (United States)

    Dando, Samantha J.; Nitsos, Ilias; Kallapur, Suhas G.; Newnham, John P.; Polglase, Graeme R.; Pillow, J. Jane; Jobe, Alan H.; Timms, Peter; Knox, Christine L.

    2012-01-01

    The multiple banded antigen (MBA) is a predicted virulence factor of Ureaplasma species. Antigenic variation of the MBA is a potential mechanism by which ureaplasmas avoid immune recognition and cause chronic infections of the upper genital tract of pregnant women. We tested whether the MBA is involved in the pathogenesis of intra-amniotic infection and chorioamnionitis by injecting virulent or avirulent-derived ureaplasma clones (expressing single MBA variants) into the amniotic fluid of pregnant sheep. At 55 days of gestation pregnant ewes (n = 20) received intra-amniotic injections of virulent-derived or avirulent-derived U. parvum serovar 6 strains (2×104 CFU), or 10B medium (n = 5). Amniotic fluid was collected every two weeks post-infection and fetal tissues were collected at the time of surgical delivery of the fetus (140 days of gestation). Whilst chronic colonisation was established in the amniotic fluid of animals infected with avirulent-derived and virulent-derived ureaplasmas, the severity of chorioamnionitis and fetal inflammation was not different between these groups (p>0.05). MBA size variants (32–170 kDa) were generated in vivo in amniotic fluid samples from both the avirulent and virulent groups, whereas in vitro antibody selection experiments led to the emergence of MBA-negative escape variants in both strains. Anti-ureaplasma IgG antibodies were detected in the maternal serum of animals from the avirulent (40%) and virulent (55%) groups, and these antibodies correlated with increased IL-1β, IL-6 and IL-8 expression in chorioamnion tissue (pureaplasmas are capable of MBA phase variation in vitro; however, ureaplasmas undergo MBA size variation in vivo, to potentially prevent eradication by the immune response. Size variation of the MBA did not correlate with the severity of chorioamnionitis. Nonetheless, the correlation between a maternal humoral response and the expression of chorioamnion cytokines is a novel finding. This host

  3. Diagnosis of foetal membrane ruptures: Placental alpha ...

    African Journals Online (AJOL)

    Context: Pre‑labour rupture of membranes (PROM) is a common obstetric complication which presents a diagnostic challenge, especially in equivocal ... problematic with the need to balance the risk of prematurity with that of prolonged ... array of rapid, minimally invasive tests based on biochemical markers in amniotic fluid ...

  4. Novel use of positively charged nylon transfer membranes for trapping indoleacetic acid or other small anions during efflux from plant tissues

    Science.gov (United States)

    Evans, M. L.; Hangarter, R. P.

    1993-01-01

    Positively charged nylon blotting membranes were used as an anion binding medium to trap [14C]indoleactic acid (IAA) as it exited cells at the basal ends of Coleus blumei L. stem and Zea mays L. coleoptile segments. Autoradiography was used to visualize where the [14C] that moved out of the cut ends was localized on the nylon membrane. Diffusion of [14C]IAA from the initial point of contact with the nylon membrane was minimal. Comparison of the autoradiograms with anatomical tissue prints of the cut ends of the segments was used to determine what tissues participate in IAA movement. The results of these initial studies were consistent with other reports suggesting that [14C]IAA movement was primarily associated with vascular tissues in both C. blumei stems and corn coleoptiles, but the resolution was not sufficient to identify which vascular tissues were involved in IAA transport. With further refinements, this technique could also be used for studying the movement of other small charged molecules through plant tissues.

  5. Intra-amniotic Ureaplasma parvum-Induced Maternal and Fetal Inflammation and Immune Responses in Rhesus Macaques.

    Science.gov (United States)

    Senthamaraikannan, Paranthaman; Presicce, Pietro; Rueda, Cesar M; Maneenil, Gunlawadee; Schmidt, Augusto F; Miller, Lisa A; Waites, Ken B; Jobe, Alan H; Kallapur, Suhas G; Chougnet, Claire A

    2016-11-15

     Although Ureaplasma species are the most common organisms associated with prematurity, their effects on the maternal and fetal immune system remain poorly characterized.  Rhesus macaque dams at approximately 80% gestation were injected intra-amniotically with 10 7 colony-forming units of Ureaplasma parvum or saline (control). Fetuses were delivered surgically 3 or 7 days later. We performed comprehensive assessments of inflammation and immune effects in multiple fetal and maternal tissues.  Although U. parvum grew well in amniotic fluid, there was minimal chorioamnionitis. U. parvum colonized the fetal lung, but fetal systemic microbial invasion was limited. Fetal lung inflammation was mild, with elevations in CXCL8, tumor necrosis factor (TNF) α, and CCL2 levels in alveolar washes at day 7. Inflammation was not detected in the fetal brain. Significantly, U. parvum decreased regulatory T cells (Tregs) and activated interferon γ production in these Tregs in the fetus. It was detected in uterine tissue by day 7 and induced mild inflammation and increased expression of connexin 43, a gap junction protein involved with labor.  U. parvum colonized the amniotic fluid and caused uterine inflammation, but without overt chorioamnionitis. It caused mild fetal lung inflammation but had a more profound effect on the fetal immune system, decreasing Tregs and polarizing them toward a T-helper 1 phenotype. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  6. INFLUENCE OF AMNIOTIC FLUID INDEX ON FOETAL OUTCOME

    Directory of Open Access Journals (Sweden)

    Raja Lakshmi

    2015-03-01

    Full Text Available BACKGROUND AND OBJECTIVE: In these days of smaller families and the obstetrician having to share the onus of giving a healthy child capable of independent existence as well as to ensure that the population is limited for further progress of this developing country, the estimation o f foetal maturity assumes greatest practical importance. As means to achieving the end, estimates of foetal maturity have been done by various clinical and laboratory methods of which assessment of amniotic fluid index assumes importance. The objective is to study the correlation of amniotic fluid index on foetal outcome at term gestation . MATERIALS AND METHODS: The study was carried out on two hundred antenatal women who attended the institute of obstetrics and gynaecology at a Government Hospital for Wome n and Children in Visakhapatnam from Jan 2014 to Jan 2015. It is a comparative prospective study comparing 100 cases of Oligohydramnios (amniotic fluid index 5 cm as control group. RESU LTS : Perinatal outcome was inferred by noting the various parameters and Statistical Analysis was done by applying the chisquare (x2 test and the value of probability was taken . CONCLUSION: The goal of antepartum fetal surveillance is to identify the fetu s at increased risk. Amniotic fluid volume has been proved as an indirect measure of feto - placental function and hence the estimation of amniotic fluid volume assists the obstetrician in risk assessment

  7. Prenatal diagnosis of Bartter syndrome: amniotic fluid aldosterone.

    Science.gov (United States)

    Rachid, Myriam; Dreux, Sophie; Pean de Ponfilly, Gauthier; Vargas-Poussou, Rosa; Czerkiewicz, Isabelle; Chevenne, Didier; Oury, Jean-François; Deschênes, Georges; Muller, Françoise

    2017-04-01

    Bartter syndrome is a severe inherited tubulopathy characterized at birth by salt wasting, severe polyuria, dehydration, growth retardation and secondary hyperaldosteronism. Prenatally, the disease is usually discovered following onset of severe polyhydramnios. We studied amniotic fluid aldosterone concentration in cases of Bartter syndrome and in control groups. Amniotic fluid aldosterone was assayed by radioimmunoassay. We undertook a retrospective case-control study based on 36 cases of postnatally diagnosed Bartter syndrome and 144 controls matched for gestational age. Two controls groups were defined: controls with polyhydramnios (n=72) and control without polyhydramnios (n=72). Amniotic fluid aldosterone was compared between the three groups. The median amniotic fluid aldosterone concentration in the Bartter syndrome group (90 pg/mL) did not differ significantly from that in the controls with polyhydramnios (90 pg/mL, p=0.33) or the controls without polyhydramnios (87 pg/mL, p=0.41). In conclusion, amniotic fluid aldosterone assay cannot be used for prenatal diagnosis of Bartter syndrome.

  8. Development and function of membrane systems in plant tissue. Annual technical progress report, 15 September 1981-15 August 1982

    International Nuclear Information System (INIS)

    Hanson, J.B.

    1982-01-01

    Over the past 11 months we have continued investigation of ion transport mechanisms in corn roots and mitochondria. In mitochondria we find that only citrate and isocitrate are transported by the H + /citrate symporter. However, the in vivo function of this carrier remains in doubt because citrate does not appear to be an effective substrate for corn mitochondria. Studies with roots have been directed to why various types of injury or shock all result in temporary blockage of the H + -efflux pump in the plasmamembrane. It appears this may be due to an injury-mediated Ca 2 + influx into the tissue, which by raising free Ca 2 + in the cytosal activates calmodulin (CaM). In turn, the Ca.CaM complex appears to activate protein kinase, phosphorylating membrane proteins. It is possible that one of these phosphorylated proteins is responsible for inactivation of the H + -ATPase. Future work is planned around the consequences of Ca 2 + influx into the root cell subsequent to injury, investigating the recovery of the H + -ATPase and the initiation of the biosyntheses which lead to augmented ion transport

  9. Gamma-ray-induced changes in hypodermal mesocarp tissue plasma membrane of pre- and post-storage muskmelon

    International Nuclear Information System (INIS)

    Lester, G.E.; Whitaker, B.D.

    1996-01-01

    Gamma irradiation (1.0 kGy) of intact, newly harvested, mature muskmelon fruit (Cucumis melo L. var. reticulatus Naud.) appears to have an immediate deleterious effect, but also a long-term beneficial effect, on the integrity and function of the plasma membrane (PM) of hypodermal mesocarp tissue. The initial consequences of gamma irradiation included an increase in the free sterol:phospholipid ratio, resulting at least in part from deglycosylation of steryl glycosides, a decrease in the spinasterol:7-stigmastenol ratio in each of the PM steryl lipids (free sterols, steryl glycosides, and acylated steryl glycosides), and a decrease in H+-ATPase activity. Irradiation did not increase protein loss, suggesting that the decrease in H+-ATPase activity resulted from either direct inactivation of the enzyme or altered PM ordering caused by the steryl lipid modifications. The long-term beneficial effects of irradiation, observed following 10 days of commercial storage, included greater retention of total PM protein, a diminished decline in total PM phospholipids (PL) and in the PL:protein ratio, and maintenance of greater overall H-ATPase activity (activity was the same as in controls on a per mg protein basis, but there was gt 30% more protein in the PM of stored irradiated fruit). These results indicate that 1 kGy gamma irradiation administered prior to storage slowes the progression of two key parameters of senescence, PM protein loss and PL catabolism

  10. Construction and characterization of an electrospun tubular scaffold for small-diameter tissue-engineered vascular grafts: a scaffold membrane approach.

    Science.gov (United States)

    Hu, Jin-Jia; Chao, Wei-Chih; Lee, Pei-Yuan; Huang, Chih-Hao

    2012-09-01

    Based on a postulate that the microstructure of a scaffold can influence that of the resulting tissue and hence its mechanical behavior, we fabricated a small-diameter tubular scaffold (∼3 mm inner diameter) that has a microstructure similar to the arterial media using a scaffold membrane approach. Scaffold membranes that contain randomly oriented, moderately aligned, or highly aligned fibers were fabricated by collecting electrospun poly([epsilon]-caprolactone) fibers on a grounded rotating drum at three different drum rotation speeds (250, 1000, and 1500 rpm). Membranes of each type were wrapped around a small-diameter mandrel to form the tubular scaffolds. Particularly, the tubular scaffolds with three different off-axis fiber angles (30, 45, and 60 degree) were formed using membranes that contain aligned fibers. These scaffolds were subjected to biaxial mechanical testing to examine the effects of fiber directions as well as the distribution of fiber orientations on their mechanical properties. The circumferential elastic modulus of the tubular scaffold was closely related to the fiber directions; the larger the off-axis fiber angle the greater the circumferential elastic modulus. The distribution of fiber orientations, on the other hand, manifested itself in the mechanical behavior via the Poisson effect. Similar to cell sheet-based vascular tissue engineering, tubular cell-seeded constructs were prepared by wrapping cell-seeded scaffold membranes, alleviating the difficulty associated with cell seeding in electrospun scaffolds. Histology of the construct illustrated that cells were aligned to the fiber directions in the construct, demonstrating the potential to control the microstructure of tissue-engineered vascular grafts using the electrospun scaffold membrane. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. 21 CFR 862.1455 - Lecithin/sphingomyelin ratio in amniotic fluid test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Lecithin/sphingomyelin ratio in amniotic fluid... Clinical Chemistry Test Systems § 862.1455 Lecithin/sphingomyelin ratio in amniotic fluid test system. (a) Identification. A lecithin/sphingomyelin ratio in amniotic fluid test system is a device intended to measure the...

  12. A rare combination of amniotic constriction band with osteogenesis imperfecta.

    Science.gov (United States)

    Shah, Krupa Hitesh; Shah, Hitesh

    2015-11-11

    Amniotic constriction bands and osteogenesis imperfecta are disorders arising from a collagen defect. We report a rare association of amniotic bands with osteogenesis imperfecta in a child. The child was born with multiple amniotic bands involving the right leg, both hands and both feet. Multiple fractures of long bones of lower limbs occurred in childhood due to trivial trauma. Deformities of the femur and tibia due to malunion with osteopenia and blue sclerae were present. The patient was treated with z plasty of constriction band of the right tibia and bisphosphonate for osteogenesis imperfecta. This rare association of both collagen diseases may provide further insight for the pathogenesis of these diseases. 2015 BMJ Publishing Group Ltd.

  13. Streptococcus agalactiae in pregnancies complicated by preterm prelabor rupture of membranes.

    Science.gov (United States)

    Musilova, Ivana; Pliskova, Lenka; Kutova, Radka; Jacobsson, Bo; Paterova, Pavla; Kacerovsky, Marian

    2016-01-01

    The main aim of this study was to evaluate the presence of Streptococcus agalactiae (S. agalactiae) in the vagina and the amniotic fluid in pregnancies complicated by preterm prelabor rupture of membranes (PPROM). The next aim was to evaluate the incidence of S. agalactiae early onset sepsis in newborns from PPROM pregnancies, with respect to the presence of S. agalactiae in the vagina and the amniotic fluid. Singleton gestations with PPROM between 24 + 0 and 36 + 6 were included. A vaginal swab was obtained, and amniocentesis was performed at admission. The presence of S. agalactiae in the vagina and in the amniotic fluid was assessed by culture and by real-time polymerase chain reaction, respectively. In total, 336 women were included. The presence of S. agalactiae in the vaginal and amniotic fluid was found in 9% (31/336) and 1% (3/336) of women. One woman had S. agalactiae in the amniotic fluid but was negative for the presence of S. agalactiae in the vaginal fluid. Early onset neonatal sepsis developed in one newborn from pregnancies complicated by the presence of S. agalactiae in the amniotic fluid. The presence of S. agalactiae in the vagina and amniotic fluid complicated approximately each 10th and each 100th PPROM pregnancy. Cultivation-negative findings of S. agalactiae in the vagina did not exclude the positivity of the amniotic fluid for S. agalactiae and the development of early onset sepsis in newborns.

  14. Histomorphometric Analysis of Periodontal Tissue Regeneration by the Use of High Density Polytetrafluoroethylen Membrane in Grade II Furcation Defects of Dogs

    Directory of Open Access Journals (Sweden)

    Raoofi S

    2015-09-01

    Full Text Available Statement of Problem: There are limited histomorphometric studies on biologic efficacy of high density tetrafluoroethylen (d-PTFE membrane. Objectives: To investigate the healing of surgically induced grade II furcation defects in dogs following the use of dense polytetrafluoroethylene as the barrier membrane and to compare the results with the contra lateral control teeth without the application of any membrane. Materials andMethods: Mandibular and maxillary 3rd premolar teeth of 18 young adult male mongrel dogs were used for the experiment. The furcation defects were created during the surgery. 5 weeks later, regenerative surgery was performed. The third premolar teeth were assigned randomly to control and test groups. In the test group, after a full thickness flap reflection, the d-PTFE membrane was placed over furcation defects. In the control group, no membrane was placed over the defect. 37 tissue blocks containing the teeth and surrounding hard and soft tissues were obtained three months post-regenerative surgery. The specimens were demineralized, serially sectioned, mounted and stained with Hematoxylin and Eosin staining technique. From each tissue block, 35-45 sections of 10 μm thickness within 60μm interval captured the entire surgically created defect. The histological images were transferred to computer and then the linear measurement ranges of the defects area, interadicular alveolar bone, epithelial attachment and coronal extension of the new cementum were done. Then, the volume and area of aforementioned parameters were calculated considering the thickness and interval of the sections. To compare the parameters between the control and test teeth, we calculated the amount of each one proportionally to the original amount of defects. Results: The mean interradicular root surface areas of original defects covered with new cementum was 74.46% and 29.59% for the membrane and control defects, respectively (p < 0.0001. Corresponding

  15. Hydrolytic Degradation and Mechanical Stability of Poly(ε-Caprolactone)/Reduced Graphene Oxide Membranes as Scaffolds for In Vitro Neural Tissue Regeneration.

    Science.gov (United States)

    Sánchez-González, Sandra; Diban, Nazely; Urtiaga, Ane

    2018-03-05

    The present work studies the functional behavior of novel poly(ε-caprolactone) (PCL) membranes functionalized with reduced graphene oxide (rGO) nanoplatelets under simulated in vitro culture conditions (phosphate buffer solution (PBS) at 37 °C) during 1 year, in order to elucidate their applicability as scaffolds for in vitro neural regeneration. The morphological, chemical, and DSC results demonstrated that high internal porosity of the membranes facilitated water permeation and procured an accelerated hydrolytic degradation throughout the bulk pathway. Therefore, similar molecular weight reduction, from 80 kDa to 33 kDa for the control PCL, and to 27 kDa for PCL/rGO membranes, at the end of the study, was observed. After 1 year of hydrolytic degradation, though monomers coming from the hydrolytic cleavage of PCL diffused towards the PBS medium, the pH was barely affected, and the rGO nanoplatelets mainly remained in the membranes which envisaged low cytotoxic effect. On the other hand, the presence of rGO nanomaterials accelerated the loss of mechanical stability of the membranes. However, it is envisioned that the gradual degradation of the PCL/rGO membranes could facilitate cells infiltration, interconnectivity, and tissue formation.

  16. Treatment of PPROM with anhydramnion in humans: first experience with different amniotic fluid substitutes for continuous amnioinfusion through a subcutaneously implanted port system.

    Science.gov (United States)

    Tchirikov, Michael; Bapayeva, Gauri; Zhumadilov, Zhaxybay Sh; Dridi, Yasmina; Harnisch, Ralf; Herrmann, Angelika

    2013-11-01

    This study aims to treat patients with preterm premature rupture of the membranes (PPROM) and anhydramnion using continuous amnioinfusion through a subcutaneously implanted port system. An amniotic fluid replacement port system was implanted in seven patients with PPROM and anhydramnion starting at the 20th week of gestation (range, 14-26 weeks) for long-term amnioinfusion. Saline solutions (2 L/day; Jonosteril(®), Sterofundin(®), isotonic NaCl 0.9% solution, lactated Ringer's solution) and a hypotonic aqueous composition with reduced chloride content similar to the electrolyte concentration of human amniotic fluid were used for the continuous amnioinfusion. The mean duration of the PPROM delivery interval continued for 49 days (range, 9-69 days), with 3 weeks of amnioinfusion via the port system (range, 4-49). The newborns showed no signs of lung hypoplasia. Long-term lavage of the amniotic cavity via a subcutaneously implanted port system in patients with PPROM and anhydramnion may help prolong the pregnancy and avoid fetal lung hypoplasia. A hypotonic aqueous composition with reduced chloride content similar to human amniotic fluid can be safely used for amnioinfusion. Prospective randomized studies are ongoing.

  17. Conformational changes and translocation of tissue-transglutaminase to the plasma membranes: role in cancer cell migration

    International Nuclear Information System (INIS)

    Kumar, Ambrish; Hu, Jianjun; LaVoie, Holly A; Walsh, Kenneth B; DiPette, Donald J; Singh, Ugra S

    2014-01-01

    Tissue-transglutaminase (TG2), a dual function G-protein, plays key roles in cell differentiation and migration. In our previous studies we reported the mechanism of TG2-induced cell differentiation. In present study, we explored the mechanism of how TG2 may be involved in cell migration. To study the mechanism of TG2-mediated cell migration, we used neuroblastoma cells (SH-SY5Y) which do not express TG2, neuroblastoma cells expressing exogenous TG2 (SHY TG2 ), and pancreatic cancer cells which express high levels of endogenous TG2. Resveratrol, a natural compound previously shown to inhibit neuroblastoma and pancreatic cancer in the animal models, was utilized to investigate the role of TG2 in cancer cell migration. Immunofluorescence assays were employed to detect expression and intracellular localization of TG2, and calcium levels in the migrating cells. Native gel electrophoresis was performed to analyze resveratrol-induced cellular distribution and conformational states of TG2 in migrating cells. Data are presented as the mean and standard deviation of at least 3 independent experiments. Comparisons were made among groups using one-way ANOVA followed by Tukey-Kramer ad hoc test. TG2 containing cells (SHY TG2 and pancreatic cancer cells) exhibit increased cell migration and invasion in collagen-coated and matrigel-coated transwell plate assays, respectively. Resveratrol (1 μM-10 μM) prevented migration of TG2-expressing cells. During the course of migration, resveratrol increased the immunoreactivity of TG2 without affecting the total TG2 protein level in migrating cells. In these cells, resveratrol increased calcium levels, and depletion of intracellular calcium by a calcium chelator, BAPTA, attenuated resveratrol-enhanced TG2 immunoreactivity. In native-polyacrylamide gels, we detected an additional TG2 protein band with slower migration in total cell lysates of resveratrol treated cells. This TG2 form is non-phosphorylated, exclusively present in plasma

  18. Intrinsic factor in human amniotic fluid as determined by radioimmunoassay

    International Nuclear Information System (INIS)

    Wahlstedt, V.; Stenman, U.-H.; Ylinen, K.; Graesbeck, R.

    1983-01-01

    The intrinsic factor (IF) concentration in 55 human amniotic fluid specimens was determined by radioimmunoassay (RIA). The antiserum was produced by immunizing rabbits with the cobalamin-IF complex isolated from human gastric juice. The median concentration of IF was 0.17 nmol/l and the extreme values <0.07-2.51 nmol/l. Three specimens with a clearly elevated level (0.96, 1.11 and 2.51 nmol/l) were observed. The highest value was associated with a fetal malformation, viz. obstruction of the proximal gut. There was no evident correlation between the concentration of IF in amniotic fluid and gestational age. (author)

  19. Utilization of ionizing radiation for the sterilization of membraneous connective tissue allografts. Part of a coordinated programme on radiation sterilization of medical products and biological tissues

    International Nuclear Information System (INIS)

    Triantafyllou, N.

    1978-05-01

    The work carried out, covered several aspects of radiation sterilization of membraneous allografts. In general the findings show that the sterilization of biological membranes with gamma irradiation using doses of up to 3 Mrads is a very satisfactory method for the sterilization of the material, since it does not seriously alter the biological properties of allografts. As for the changes of the mechanical properties, this is not considered to be a serious draw back, because the membraneous allografts cover surgical needs where the mechanical strength is not of paramount importance

  20. Yolk sac development in lizards (Lacertilia: Scincidae): New perspectives on the egg of amniotes.

    Science.gov (United States)

    Stewart, James R; Thompson, Michael B

    2017-04-01

    Embryos of oviparous reptiles develop on the surface of a large mass of yolk, which they metabolize to become relatively large hatchlings. Access to the yolk is provided by tissues growing outward from the embryo to cover the surface of the yolk. A key feature of yolk sac development is a dedicated blood vascular system to communicate with the embryo. The best known model for yolk sac development and function of oviparous amniotes is based on numerous studies of birds, primarily domestic chickens. In this model, the vascular yolk sac forms the perimeter of the large yolk mass and is lined by a specialized epithelium, which takes up, processes and transports yolk nutrients to the yolk sac blood vessels. Studies of lizard yolk sac development, dating to more than 100 years ago, report characteristics inconsistent with this model. We compared development of the yolk sac from oviposition to near hatching in embryonic series of three species of oviparous scincid lizards to consider congruence with the pattern described for birds. Our findings reinforce results of prior studies indicating that squamate reptiles mobilize and metabolize the large yolk reserves in their eggs through a process unknown in other amniotes. Development of the yolk sac of lizards differs from birds in four primary characteristics, migration of mesoderm, proliferation of endoderm, vascular development and cellular diversity within the yolk sac cavity. Notably, all of the yolk is incorporated into cells relatively early in development and endodermal cells within the yolk sac cavity align along blood vessels which course throughout the yolk sac cavity. The pattern of uptake of yolk by endodermal cells indicates that the mechanism of yolk metabolism differs between lizards and birds and that the evolution of a fundamental characteristic of embryonic nutrition diverged in these two lineages. Attributes of the yolk sac of squamates reveal the existence of phylogenetic diversity among amniote lineages

  1. Early fish myoseptal cells: insights from the trout and relationships with amniote axial tenocytes.

    Directory of Open Access Journals (Sweden)

    Yoann Bricard

    Full Text Available The trunk muscle in fish is organized as longitudinal series of myomeres which are separated by sheets of connective tissue called myoseptum to which myofibers attach. In this study we show in the trout that the myoseptum separating two somites is initially acellular and composed of matricial components such as fibronectin, laminin and collagen I. However, myoseptal cells forming a continuum with skeletogenic cells surrounding axial structures are observed between adjacent myotomes after the completion of somitogenesis. The myoseptal cells do not express myogenic markers such as Pax3, Pax7 and myogenin but express several tendon-associated collagens including col1a1, col5a2 and col12a1 and angiopoietin-like 7, which is a secreted molecule involved in matrix remodelling. Using col1a1 as a marker gene, we observed in developing trout embryo an initial labelling in disseminating cells ventral to the myotome. Later, labelled cells were found more dorsally encircling the notochord or invading the intermyotomal space. This opens the possibility that the sclerotome gives rise not only to skeletogenic mesenchymal cells, as previously reported, but also to myoseptal cells. We furthermore show that myoseptal cells differ from skeletogenic cells found around the notochord by the specific expression of Scleraxis, a distinctive marker of tendon cells in amniotes. In conclusion, the location, the molecular signature and the possible sclerotomal origin of the myoseptal cells suggest that the fish myoseptal cells are homologous to the axial tenocytes in amniotes.

  2. Choriodecidual group B streptococcal inoculation induces fetal lung injury without intra-amniotic infection and preterm labor in Macaca nemestrina.

    Directory of Open Access Journals (Sweden)

    Kristina M Adams Waldorf

    Full Text Available BACKGROUND: Early events leading to intrauterine infection and fetal lung injury remain poorly defined, but may hold the key to preventing neonatal and adult chronic lung disease. Our objective was to establish a nonhuman primate model of an early stage of chorioamnionitis in order to determine the time course and mechanisms of fetal lung injury in utero. METHODOLOGY/PRINCIPAL FINDINGS: Ten chronically catheterized pregnant monkeys (Macaca nemestrina at 118-125 days gestation (term=172 days received one of two treatments: 1 choriodecidual and intra-amniotic saline (n=5, or 2 choriodecidual inoculation of Group B Streptococcus (GBS 1×10(6 colony forming units (n=5. Cesarean section was performed regardless of labor 4 days after GBS or 7 days after saline infusion to collect fetal and placental tissues. Only two GBS animals developed early labor with no cervical change in the remaining animals. Despite uterine quiescence in most cases, blinded review found histopathological evidence of fetal lung injury in four GBS animals characterized by intra-alveolar neutrophils and interstitial thickening, which was absent in controls. Significant elevations of cytokines in amniotic fluid (TNF-α, IL-8, IL-1β, IL-6 and fetal plasma (IL-8 were detected in GBS animals and correlated with lung injury (p<0.05. Lung injury was not directly caused by GBS, because GBS was undetectable in amniotic fluid (~10 samples tested/animal, maternal and fetal blood by culture and polymerase chain reaction. In only two cases was GBS cultured from the inoculation site in low numbers. Chorioamnionitis occurred in two GBS animals with lung injury, but two others with lung injury had normal placental histology. CONCLUSIONS/SIGNIFICANCE: A transient choriodecidual infection can induce cytokine production, which is associated with fetal lung injury without overt infection of amniotic fluid, chorioamnionitis or preterm labor. Fetal lung injury may, thus, occur silently without

  3. The Use of Fibrous, Supramolecular Membranes and Human Tubular Cells for Renal Epithelial Tissue Engineering : Towards a Suitable Membrane for a Bioartificial Kidney

    NARCIS (Netherlands)

    Dankers, Patricia Y. W.; Boomker, Jasper M.; Huizinga-van der Vlag, Ali; Smedts, Frank M. M.; Harmsen, Martin C.; van Luyn, Marja J. A.

    2010-01-01

    A bioartificial kidney, which is composed of a membrane cartridge with renal epithelial cells, can substitute important kidney functions in patients with renal failure. A particular challenge is the maintenance of monolayer integrity and specialized renal epithelial cell functions ex vivo. We

  4. The use of fibrous, supramolecular membranes and human tubular cells for renal epithelial tissue engineering: towards a suitable membrane for a bioartificial kidney,

    NARCIS (Netherlands)

    Dankers, P.Y.W.; Boomker, J.M.; Huizinga-van der Vlag, A.; Smedts, F.M.M.; Harmsen, M.C.; Luyn, van M.J.A.

    2010-01-01

    A bioartificial kidney, which is composed of a membrane cartridge with renal epithelial cells, can substitute important kidney functions in patients with renal failure. A particular challenge is the maintenance of monolayer integrity and specialized renal epithelial cell functions ex vivo. We

  5. Production and Characterization of a Novel, Electrospun, Tri-Layer Polycaprolactone Membrane for the Segregated Co-Culture of Bone and Soft Tissue

    Directory of Open Access Journals (Sweden)

    Sasima Puwanun

    2016-06-01

    Full Text Available Composite tissue-engineered constructs combining bone and soft tissue have applications in regenerative medicine, particularly dentistry. This study generated a tri-layer, electrospun, poly-ε-caprolactone membrane, with two microfiber layers separated by a layer of nanofibers, for the spatially segregated culture of mesenchymal progenitor cells (MPCs and fibroblasts. The two cell types were seeded on either side, and cell proliferation and spatial organization were investigated over several weeks. Calcium deposition by MPCs was detected using xylenol orange (XO and the separation between fibroblasts and the calcified matrix was visualized by confocal laser scanning microscopy. SEM confirmed that the scaffold consisted of two layers of micron-diameter fibers with a thin layer of nano-diameter fibers in-between. Complete separation of cell types was maintained and calcified matrix was observed on only one side of the membrane. This novel tri-layer membrane is capable of supporting the formation of a bilayer of calcified and non-calcified connective tissue.

  6. Amniotic band-like structures | Govender | Obstetrics and ...

    African Journals Online (AJOL)

    Intra-amniotic band-like structures are seen fairly commonly on routine obstetric scans, especially during the first and second trimesters of pregnancy. It is important to establish the cause for such findings in order to determine their clinical significance and to assess prognosis. The vast majority of band-like structures are ...

  7. Amniotic fluid as a source of engraftable stem cells

    Directory of Open Access Journals (Sweden)

    Cesar V Borlongan

    2017-01-01

    Full Text Available The ability of stem cells to differentiate into various lineages has made them powerful tools of regenerative medicine and applicable to multiple human diseases. Of particular interest, amniotic fluid-derived stem cells (AFSC have been characterized to express both adult and embryonic cell markers, indicating them as cells within an intermediate stage between embryonic and adult phenotype. AFSC can differentiate into cells of all three germ layers, including hepatic, myogenic, osteogenic, and neurogenic cell types. Furthermore, AFSC have minimal replicative senescence, retaining the ability to divide effectively for over 250 doublings. These facts indicate that amniotic fluid may exist as a promising donor source of stem cells for the treatment of multiple clinically relevant conditions. Of particular interest is the convenience of harvesting stem cells from the amniotic fluid stem for the treatment of newborns, as well as for banking or cryopreserving purposes to be used at a later date. Importantly, the promise of amniotic fluid as a source of stem cells merits ongoing research into their potential therapeutic applications. This paper is a review article. Referred literature in this paper has been listed in the references section. The datasets supporting the conclusions of this article are available online by searching various databases, including PubMed. Some original points in this article come from the laboratory practice in our research center and the authors' experiences.

  8. Amniotic fluid MMP-9 and neurotrophins in autism spectrum disorders

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Pearce, Brad D; Larsen, Nanna

    2012-01-01

    Evidence suggests that some developmental disorders, such as autism spectrum disorders (ASDs), are caused by errors in brain plasticity. Given the important role of matrix metalloproteinases (MMPs) and neurotrophins (NTs) in neuroplasticity, amniotic fluid samples for 331 ASD cases and 698...

  9. Amniotic fluid index predicts the relief of variable decelerations after amnioinfusion bolus.

    Science.gov (United States)

    Spong, C Y; McKindsey, F; Ross, M G

    1996-10-01

    Our purpose was to determine whether intrapartum amniotic fluid index before amnioinfusion can be used to predict response to therapeutic amnioinfusion. Intrapartum patients (n = 85) with repetitive variable decelerations in fetal heart rate that necessitated amnioinfusion (10 ml/min for 60 minutes) underwent determination of amniotic fluid index before and after bolus amnioinfusion. The fetal heart tracing was scored (scorer blinded to amniotic fluid index values) for number and characteristics of variable decelerations before and 1 hour after initiation of amnioinfusion. The amnioinfusion was considered successful if it resulted in a decrease of > or = 50% in total number of variable decelerations or a decrease of > or = 50% in the rate of atypical or severe variable decelerations after administration of the bolus. Spontaneous vaginal births before completion of administration of the bolus (n = 18) were excluded from analysis. The probability of success of amnioinfusion in relation to amniotic fluid index was analyzed with the chi(2) test for progressive sequence. The mean amniotic fluid index before amnioinfusion was 6.2 +/- 3.3 cm. An amniotic fluid index of amnioinfusion decreased with increasing amniotic fluid index before amnioinfusion (76% [16/21] when initial amniotic fluid index was 0 to 4 cm, 63% [17/27] when initial amniotic fluid index was 4 to 8 cm, 44% [7/16] when initial amniotic fluid index was 8 to 12 cm, and 33% [1/3] when initial amniotic fluid index was > 12 cm, p = 0.03). The incidence of nuchal cords or true umbilical cord knots increased in relation to amniotic fluid index before amnioinfusion. Amniotic fluid index before amnioinfusion can be used to predict the success of amnioinfusion for relief of variable decelerations in fetal heart rate. Failure of amnioinfusion at a high amniotic fluid index before amnioinfusion may be explained by the increased prevalence of nuchal cords or true knots in the umbilical cord.

  10. The tissue-specific Rep8/UBXD6 tethers p97 to the endoplasmic reticulum membrane for degradation of misfolded proteins.

    Directory of Open Access Journals (Sweden)

    Louise Madsen

    Full Text Available The protein known as p97 or VCP in mammals and Cdc48 in yeast is a versatile ATPase complex involved in several biological functions including membrane fusion, protein folding, and activation of membrane-bound transcription factors. In addition, p97 plays a central role in degradation of misfolded secretory proteins via the ER-associated degradation pathway. This functional diversity of p97 depends on its association with various cofactors, and to further our understanding of p97 function it is important that these cofactors are identified and analyzed. Here, we isolate and characterize the human protein named Rep8 or Ubxd6 as a new cofactor of p97. Mouse Rep8 is highly tissue-specific and abundant in gonads. In testes, Rep8 is expressed in post-meiotic round spermatids, whereas in ovaries Rep8 is expressed in granulosa cells. Rep8 associates directly with p97 via its UBX domain. We show that Rep8 is a transmembrane protein that localizes to the ER membrane with its UBX domain facing the cytoplasm. Knock-down of Rep8 expression in human cells leads to a decreased association of p97 with the ER membrane and concomitantly a retarded degradation of misfolded ER-derived proteasome substrates. Thus, Rep8 tethers p97 to the ER membrane for efficient ER-associated degradation.

  11. H+-ATPase activity from storage tissue of Beta vulgaris. IV. N,N'-dicyclohexylcarbodiimide binding and inhibition of the plasma membrane H+-ATPase

    International Nuclear Information System (INIS)

    Oleski, N.A.; Bennett, A.B.

    1987-01-01

    The molecular weight and isoelectric point of the plasma membrane H + -ATPase from red beet storage tissue were determined using N,N'-dicyclohexylcarbodiimide (DCCD) and a H + -ATPase antibody. When plasma membrane vesicles were incubated with 20 micromolar [ 14 C]-DCCD at 0 0 C, a single 97,000 dalton protein was visualized on a fluorography of a sodium dodecyl sulfate polyacrylamide gel. A close correlation between [ 14 C]DCCD labeling of the 97,000 dalton protein and the extent of ATPase inhibition over a range of DCCD concentration suggests that this 97,000 dalton protein is a component of the plasma membrane H + -ATPase. An antibody raised against the plasma membrane H + -ATPase of Neurospora crassa cross-reacted with the 97,000 dalton DCCD-binding protein, further supporting the identity of this protein. Immunoblots of two-dimensional gels of red beet plasma membrane vesicles indicated the isoelectric point of the H + -ATPase to be 6.5

  12. An Adaptable Spectrin/Ankyrin-Based Mechanism for Long-Range Organization of Plasma Membranes in Vertebrate Tissues.

    Science.gov (United States)

    Bennett, Vann; Lorenzo, Damaris N

    2016-01-01

    Ankyrins are membrane-associated proteins that together with their spectrin partners are responsible for micron-scale organization of vertebrate plasma membranes, including those of erythrocytes, excitable membranes of neurons and heart, lateral membrane domains of columnar epithelial cells, and striated muscle. Ankyrins coordinate functionally related membrane transporters and cell adhesion proteins (15 protein families identified so far) within plasma membrane compartments through independently evolved interactions of intrinsically disordered sequences with a highly conserved peptide-binding groove formed by the ANK repeat solenoid. Ankyrins are coupled to spectrins, which are elongated organelle-sized proteins that form mechanically resilient arrays through cross-linking by specialized actin filaments. In addition to protein interactions, cellular targeting and assembly of spectrin/ankyrin domains also critically depend on palmitoylation of ankyrin-G by aspartate-histidine-histidine-cysteine 5/8 palmitoyltransferases, as well as interaction of beta-2 spectrin with phosphoinositide lipids. These lipid-dependent spectrin/ankyrin domains are not static but are locally dynamic and determine membrane identity through opposing endocytosis of bulk lipids as well as specific proteins. A partnership between spectrin, ankyrin, and cell adhesion molecules first emerged in bilaterians over 500 million years ago. Ankyrin and spectrin may have been recruited to plasma membranes from more ancient roles in organelle transport. The basic bilaterian spectrin-ankyrin toolkit markedly expanded in vertebrates through gene duplications combined with variation in unstructured intramolecular regulatory sequences as well as independent evolution of ankyrin-binding activity by ion transporters involved in action potentials and calcium homeostasis. In addition, giant vertebrate ankyrins with specialized roles in axons acquired new coding sequences by exon shuffling. We speculate that

  13. Characterization of an apically derived epithelial membrane glycoprotein from bovine milk, which is expressed in capillary endothelia in diverse tissues

    OpenAIRE

    1985-01-01

    A glycoprotein (PAS IV) of apparent Mr 76,000 was purified from bovine milk-fat-globule membrane and partially characterized. PAS IV contained mannose, galactose, and sialic acid as principal sugars (approximately 5.3% total carbohydrate [wt/wt]) and existed in milk in at least four isoelectric variants. The glycoprotein appeared to be an integral membrane protein by several criteria. PAS IV was recovered in the detergent phase of Triton X-114 extracts of milk-fat-globule membrane at room tem...

  14. Activity of adenylate cyclase in plasma membranes of pulmonary tissue remote times following nonlethal gamma-irradiation of rats

    International Nuclear Information System (INIS)

    Slozhenkina, L.V.; Ruda, V.P.; Ushakova, T.E.; Kuzin, A.M.

    1990-01-01

    Basal and stimulated activity of adenylate cyclase (cyclizing ATP-pyrophosphate lyase, E.C. 4.6.1.1., AC) in plasma membranes of pumonary tissye was being studied during a year after fractionated irradiation of rats (2 Gyx3). Basal and hormone-stimulated activity of AC was shown to vary significantly from normal 6 and 12 months after irradiation. The exposed membranes responded differently to AC activation by isoproterenol and F -

  15. Fibrous guided tissue regeneration membrane loaded with anti-inflammatory agent prepared by coaxial electrospinning for the purpose of controlled release

    Energy Technology Data Exchange (ETDEWEB)

    He, Min; Xue, Jiajia [Beijing Laboratory of Biomedical Materials, Beijing University of Chemical Technology, Beijing 100029 (China); Geng, Huan; Gu, Hao [State Key Laboratory of Organic–Inorganic Composites, Beijing University of Chemical Technology, Beijing 100029 (China); Chen, Dafu [Laboratory of Bone Tissue Engineering of Beijing Research Institute of Traumatology and Orthopaedics, Beijing 100035 (China); Shi, Rui, E-mail: sharell@126.com [Laboratory of Bone Tissue Engineering of Beijing Research Institute of Traumatology and Orthopaedics, Beijing 100035 (China); Zhang, Liqun, E-mail: zhanglq@mail.buct.edu.cn [Beijing Laboratory of Biomedical Materials, Beijing University of Chemical Technology, Beijing 100029 (China); State Key Laboratory of Organic–Inorganic Composites, Beijing University of Chemical Technology, Beijing 100029 (China)

    2015-04-30

    Graphical abstract: The metronidazole released from PCL/gelatin core/sheath nanofiber membranes can effectively inhibit the colonization of anerobic bacteria. - Highlights: • Core/sheath PCL/gelatin nanofiber membrane loaded with metronidazole in a wide range of drug loading (5–35 wt.%) were successfully fabricated in good quality. • The encapsulation of gelatin can effectively alleviate the initial burst release of drugs. • The membrane can inhibit the growth of bacteria as the drug content reaches 10% (w/w), and the bacterial inhibition ability can effectively last at least 4 weeks. • The encapsulation of gelatin can overcome the disadvantage of PCL's hydrophobicity, which can effectively promote the adhesion and proliferation of cells. - Abstract: Here, with the aim of inhibiting inflammation during guided tissue regeneration membrane (GTRM) implant surgery, coaxial electrospinning was used to fabricate drug-loaded core/sheath nanofiber GTRMs capable of controlled drug release. Various amounts of the anti-inflammatory agent metronidazole (MNA) were encapsulated into the core/sheath nanofibers (where PCL was the core, gelatin the sheath, and the gelatin shell was crosslinked with genipin) in order to establish the minimal drug content necessary to achieve the appropriate anti-inflammatory effect. By using TEM and SEM, the core/sheath structure was confirmed. In vitro drug disolution results showed that the core/sheath nanofibers exhibited sustained release profiles that were superior to those nanofibers produced by blending electrospinning. Additionally, the membrane significantly inhibited the colonization of anaerobic bacteria. Furthermore, with gelatin as a shell, the core/shell nanofiber membranes showed improved hydrophilicity, which resulted in better cell adhesion and proliferation without cytotoxicity. Therefore, in this study, a simple and effective coaxial electrospinning approach was demonstrated for the fabrication of anti

  16. Amniotic fluid stem cells: a promising therapeutic resource for cell-based regenerative therapy.

    Science.gov (United States)

    Antonucci, Ivana; Pantalone, Andrea; Tete, Stefano; Salini, Vincenzo; Borlongan, Cesar V; Hess, David; Stuppia, Liborio

    2012-01-01

    Stem cells have been proposed as a powerful tool in the treatment of several human diseases, both for their ability to represent a source of new cells to replace those lost due to tissue injuries or degenerative diseases, and for the ability of produce trophic molecules able to minimize damage and promote recovery in the injured tissue. Different cell types, such as embryonic, fetal or adult stem cells, human fetal tissues and genetically engineered cell lines, have been tested for their ability to replace damaged cells and to restore the tissue function after transplantation. Amniotic fluid -derived Stem cells (AFS) are considered a novel resource for cell transplantation therapy, due to their high renewal capacity, the "in vitro" expression of embryonic cell lineage markers, and the ability to differentiate in tissues derived from all the three embryonic layers. Moreover, AFS do not produce teratomas when transplanted into animals and are characterized by a low antigenicity, which could represent an advantage for cell transplantation or cell replacement therapy. The present review focuses on the biological features of AFS, and on their potential use in the treatment of pathological conditions such as ischemic brain injury and bone damages.

  17. The tissue-specific Rep8/UBXD6 tethers p97 to the endoplasmic reticulum membrane for degradation of misfolded proteins

    DEFF Research Database (Denmark)

    Madsen, Louise; Kriegenburg, Franziska; Lages Lino Vala, Andrea

    2011-01-01

    is a transmembrane protein that localizes to the ER membrane with its UBX domain facing the cytoplasm. Knock-down of Rep8 expression in human cells leads to a decreased association of p97 with the ER membrane and concomitantly a retarded degradation of misfolded ER-derived proteasome substrates. Thus, Rep8 tethers p......The protein known as p97 or VCP in mammals and Cdc48 in yeast is a versatile ATPase complex involved in several biological functions including membrane fusion, protein folding, and activation of membrane-bound transcription factors. In addition, p97 plays a central role in degradation of misfolded...... protein named Rep8 or Ubxd6 as a new cofactor of p97. Mouse Rep8 is highly tissue-specific and abundant in gonads. In testes, Rep8 is expressed in post-meiotic round spermatids, whereas in ovaries Rep8 is expressed in granulosa cells. Rep8 associates directly with p97 via its UBX domain. We show that Rep8...

  18. Tissue banking in the Asia Pacific region - an overview

    International Nuclear Information System (INIS)

    Hasim Mohamad

    1999-01-01

    Tissue banking is a relative new entity in the Asia-Pacific region. Sporadic cases of tissue transplantation were reported in the early sixties whereby corneas mainly from Sri Lanka were airflown to ophthalmic surgeons in this region. Till today Sri Lanka has been playing a key role in supplying corneas and other tissues throughout the globe. Similar cases of allograft bone transplantation were reported from Myanmar during that period. However it was not until early eighties when a group of scientist medical doctors from various countries in this region met in Colombo under IAEA to discuss the possibilities of developing tissue banks in their respective countries. This development had been very slow and until today only about fourteen countries are directly involved. Due to the lack of financial support both from the authorities or non-government organizations most of the tissue banks survived by research grants from universities, the government research and development grants or grants from private bodies which are hard to come by and are usually inadequate to sustain or to maintain the banks. Most of our tissue banks started small by involving in amniotic membrane processing for use in bum patients in their initial stage of development. These are true for countries like China, Malaysia, Bangladesh, Pakistan and Indonesia where amniotic membranes are easily available and the processing cost is cheap. The availability of cobalt source for radiation of these tissues is also an important factor. Some of these banks until today still focussed on these areas whereas others had rapidly progress into production of other tissue such as bones, heart valves, allograft skin, and xenograft bones. Richer counties in the Pacific rim, where financing is not a major problem, the progress of their banks depends greatly on the demands of tissue by the clinicians and the availability of raw materials and tissue bank facilities. At the moment due to the lack of donors the raw

  19. RESEARCH ON REDUCING PREMATURITY RUPTURE OF MEMBRANE

    Directory of Open Access Journals (Sweden)

    Maria URSACHI (BOLOTA

    2016-12-01

    Full Text Available The membranes surrounding the amniotic cavity are composed from amnion and chorion, tightly adherent layers which are composed of several cell types, including epithelial cells, trophoblasts cells and mesenchyme cells, embedded in a collagenous matrix. They retain amniotic fluid, secret substances into the amniotic fluid, as well as to the uterus and protect the fetus against upward infections from urogenital tract. Normally, the membranes it breaks during labor. Premature rupture of the amniotic sac (PRAS is defined as rupture of membranes before the onset of labor. Premature rupture of the fetal membrane, which occurs before 37 weeks of gestation, usually, refers to preterm premature rupture of membranes. Despite advances in the care period, premature rupture of membranes and premature rupture of membranes preterm continue to be regarded as serious obstetric complications. On the term 8% - 10% of pregnant women have premature rupture of membranes; these women are at increased risk of intrauterine infections, where the interval between membrane rupture and expulsion is rolled-over. Premature rupture of membranes preterm occurs in approximately 1% of all pregnancies and is associated with 30% -40% of preterm births. Thus, it is important to identify the cause of pre-term birth (after less than 37 completed weeks of "gestation" and its complications, including respiratory distress syndrome, neonatal infection and intraventricular hemorrhage. Objectives: the development of the protocol of the clinical trial on patients with impending preterm birth, study clinical and statistical on the socio-demographic characteristics of patients with imminent preterm birth; clinical condition of patients and selection of cases that could benefit from the application of interventional therapy; preclinical investigation (biological and imaging of patients with imminent preterm birth; the modality therapy; clinical investigation of the effectiveness of short

  20. [Intrapartum amnioinfusion in patients with meconium-stained amniotic fluid].

    Science.gov (United States)

    Engel, Karina; Samborska, Monika; Bilar, Marek; Sipak-Szmigiel, Olimpia; Ronin-Walknowska, Elzbieta

    2008-09-01

    The aim of the study was to evaluate the effect of intrapartum amnioinfusion in the presence of meconium stained amniotic fluid. 93 women with meconium-stained amniotic fluid were assigned to receive amnioinfusion or no amnioinfusion (128 women). The trials were evaluated for fetal distress syndrome, route of delivery, fetal acidemia, Apgar score at 1 and 5 min., meconium aspiration syndrome, postpartum endometritis and maternal hospital stays. Amnioinfusion in cases of meconium-stained fluid did not improve the number of fetal distress symptoms during fetal heart rate monitoring. Amnioinfusion was associated with a significant decrease of neonatal acidemia although it did not improve Apgar score. In our study amnioinfusion was not associated with reduction in the incidence of neonatal outcome and puerperial complications.

  1. RIA of alpha-fetoprotein in serum and amniotic fluid

    International Nuclear Information System (INIS)

    Fingerova, H.; Talas, M.; Stroufova, A.

    1979-01-01

    An own modification of the double antibody radioimmunoassay for AFP using 125 I-labelled AFP as a tracer, rabbit anti-AFP obtained from SEVAC, Prague and precipitating antibodies prepared by the authors is described. The AFP levels measured in the serum and the amniotic fluid using the method were in agreement with those obtained by the means of the AFPK RIA kit by SORIN in the Institute for the Care of Mother and Child in Prague. The AFP concentrations found in the cord serum and the amniotic fluid were confirmed also by the rocket electroimmunoassay according to Laurell. The described AFP RIA seems suitable for the clinical application in prenatal screening for congenital malformations, in difficult pregnancies, in hepatology and the diagnosis and the evaluation of therapy of some human malignancies. (author)

  2. RIA of alpha-fetoprotein in serum and amniotic fluid

    Energy Technology Data Exchange (ETDEWEB)

    Fingerova, H; Talas, M; Stroufova, A [Palackeho Univ., Olomouc (Czechoslovakia). Lekarska Fakulta; Santavy, J; Krikal, Z [Ustav pro Peci o Matku a Dite, Prague (Czechoslovakia)

    1979-01-01

    An own modification of the double antibody radioimmunoassay for AFP using /sup 125/I-labelled AFP as a tracer, rabbit anti-AFP obtained from SEVAC, Prague and precipitating antibodies prepared by the authors is described. The AFP levels measured in the serum and the amniotic fluid using the method were in agreement with those obtained by the means of the AFPK RIA kit by SORIN in the Institute for the Care of Mother and Child in Prague. The AFP concentrations found in the cord serum and the amniotic fluid were confirmed also by the rocket electroimmunoassay according to Laurell. The described AFP RIA seems suitable for the clinical application in prenatal screening for congenital malformations, in difficult pregnancies, in hepatology and the diagnosis and the evaluation of therapy of some human malignancies.

  3. Comparative evaluation of a bioabsorbable collagen membrane and connective tissue graft in the treatment of localized gingival recession: A clinical study

    Directory of Open Access Journals (Sweden)

    Harsha Mysore Babu

    2011-01-01

    Full Text Available Background: Gingival recession (GR can result in root sensitivity, esthetic concern to the patient, and predilection to root caries. The purpose of this randomized clinical study was to evaluate (1 the effect of guided tissue regeneration (GTR procedure using a bioabsorbable collagen membrane, in comparison to autogenous subepithelial connective tissue graft (SCTG for root coverage in localized gingival recession defects; and (2 the change in width of keratinized gingiva following these two procedures. Materials and Methods: A total of 10 cases, showing at least two localized Miller′s Class I or Class II gingival recession, participated in this study. In a split mouth design, the pairs of defects were randomly assigned for treatment with either SCTG (SCTG Group or GTR-based collagen membrane (GTRC Group. Both the grafts were covered with coronally advanced flap. Recession depth (RD, recession width (RW, width of keratinized gingiva (KG, probing depth (PD, relative attachment level (RAL, plaque index (PI, and gingival index (GI were recorded at baseline, 3 and 6 months postoperatively. Results: Six months following root coverage procedures, the mean root coverage was found to be 84.84% ± 16.81% and 84.0% ± 15.19% in SCTG Group and GTRC Group, respectively. The mean keratinized gingival width increase was 1.50 ± 0.70 mm and 2.30 ± 0.67 mm in the SCTG and GTRC group, respectively, which was not statistically significant. Conclusion: It may be concluded that resorbable collagen membrane can be a reliable alternative to autogenous connective tissue graft in the treatment of gingival recession.

  4. The oldest record of aquatic amniote congenital scoliosis.

    Directory of Open Access Journals (Sweden)

    Tomasz Szczygielski

    Full Text Available We report the first occurrence of congenital scoliosis in an early Permian aquatic parareptile, Stereosternum tumidum from Paraná state, Brazil. The spine malformation is caused by a congenital hemivertebra. These observations give insight into the biomechanical aspects of underwater locomotion in an axial skeleton-compromised aquatic amniote. This is the oldest record of a hemivertebra in an aquatic animal.

  5. Results of six years of cytogenetic studies in amniotic fluid

    Directory of Open Access Journals (Sweden)

    Enelis Reyes Reyes

    2015-10-01

    Full Text Available Background: research into different genetic diseases is one of the preventive programs of paramount importance at public health level. The early detection of chromosomopathies and the establishment of an appropriate strategy reduce the morbidity-morality rate and improve the patients’ quality of life.Objective: to describe the behavior of the results of the cytogenetic studies in the amniotic fluid of pregnant women from Las Tunas province during six years: from 2008 to 2014.Methods: a retrospective and descriptive study was carried out to assess the results of cytogenetic studies in amniotic liquid during six years: from 2008 to 2014. The statistical records were checked and the results, the indication criteria, the behavior of the age groups in women advanced in age and the diagnosed chromosomopathies were assessed.Results: the samples with results that exceeded the non-conclusive and positive women prevailed; 2, 3 positive cases of chromosomopathies were diagnosed out of 100 studied women at risk; pregnant women of advanced gestational years prevailed as indication criterion, being the 37 to 40 years old age group the predominant one; in the positive cases, numeric chromosomopathies of the type trisomy 21 or Down’s syndrome prevailed, with a frequency of 1, 2 out of 100 pregnant women at risk.Conclusions: the program of the cytogenetic diagnosis in the amniotic fluid has been an effective tool to detect congenital prenatal defects by chromosomopathies, very useful in the process of genetic advice.

  6. Bartter syndrome prenatal diagnosis based on amniotic fluid biochemical analysis.

    Science.gov (United States)

    Garnier, Arnaud; Dreux, Sophie; Vargas-Poussou, Rosa; Oury, Jean-François; Benachi, Alexandra; Deschênes, Georges; Muller, Françoise

    2010-03-01

    Bartter syndrome is an autosomic recessive disease characterized by severe polyuria and sodium renal loss. The responsible genes encode proteins involved in electrolyte tubular reabsorption. Prenatal manifestations, mainly recurrent polyhydramnios because of fetal polyuria, lead to premature delivery. After birth, polyuria leads to life-threatening dehydration. Prenatal genetic diagnosis needs an index case. The aim of this study was to analyze amniotic fluid biochemistry for the prediction of Bartter syndrome. We retrospectively studied 16 amniotic fluids of Bartter syndrome-affected fetuses diagnosed after birth, only six of them being genetically proven. We assayed total proteins, alpha-fetoprotein, and electrolytes and defined a Bartter index corresponding to the multiplication of total protein and of alpha-fetoprotein. Results were compared with two control groups matched for gestational age-non-Bartter polyhydramnios (n = 30) and nonpolyhydramnios (n = 60). In Bartter syndrome, we observed significant differences (p Bartter index (0.16, 0.82, and 1.0, respectively). No statistical difference was observed for electrolytes. In conclusion, Bartter syndrome can be prenatally suspected on amniotic fluid biochemistry (sensitivity 93% and specificity 100%), allowing appropriate management before and after birth.

  7. Peptidome analysis of amniotic fluid from pregnancies with preeclampsia.

    Science.gov (United States)

    Qian, Yating; Zhang, Lei; Rui, Can; Ding, Hongjuan; Mao, Pengyuan; Ruan, Hongjie; Jia, Ruizhe

    2017-11-01

    Preeclampsia (PE), a life‑threatening, complicated pregnancy‑associated disease, has recently become a research focus in obstetrics. However, the peptidome of the amniotic fluid in PE patients has rarely been investigated. The present study used peptidomic profiling to perform a comparative analysis of human amniotic fluid between normal and PE pregnancies. Centrifugal ultrafiltration and liquid chromatography‑tandem mass spectrometry (LC‑MS/MS) was combined with isotopomeric dimethyl labels to gain a deeper understanding of the role of proteins and the peptidome in the onset of PE. Following ultrafiltration and LC‑MS/MS, 352 peptides were identified. Of these, 23 peptides were observed to be significantly differentially expressed (6 downregulated and 17 upregulated; POntology and Blastp analyses, the functions and biological activities of these 23 peptides were identified and revealed to include autophagy, signal transduction, receptor activity, enzymatic activity and nucleic acid binding. In addition, a bibliographic search revealed that some of the identified peptides, including Titin, are crucial to the pathogenesis underlying PE. The present study identified 23 peptides expressed at significantly different levels in the amniotic fluid of PE and normal pregnancies. A comprehensive peptidome analysis is more efficient than a simple biomarker analysis at revealing deficiencies and improving the detection rate in diseases. These analyses therefore provide a substantial advantage in applications aimed at the discovery of disease‑specific biomarkers.

  8. Tissue-specific expression of the gene for a putative plasma membrane H(+)-ATPase in a seagrass.

    Science.gov (United States)

    Fukuhara, T; Pak, J Y; Ohwaki, Y; Tsujimura, H; Nitta, T

    1996-01-01

    A cDNA clone corresponding to the gene (ZHA1) for a putative plasma membrane H(+)-ATPase of a seagrass (Zostera marina L.) was isolated and sequenced. Comparison of the amino acid predicted sequence from the nucleotide sequence of ZHA1 with those encoded by known genes for plasma membrane H(+)-ATPases from other plants indicated that ZHA1 is most similar to the gene (PMA4) for a plasma membrane H(+)-ATPase in a tobacco (84.4%). Northern hybridization indicated that ZHA1 was strongly expressed in mature leaves, which are exposed to seawater and have the ability of tolerate salinity; ZHA1 was weakly expressed in immature leaves, which are protected from seawater by tightly enveloping sheaths and are sensitive to salinity. In mature leaves, in situ hybridization revealed that ZHA1 was expressed specifically in epidermal cells, the plasma membranes of which were highly invaginated and morphologically similar to those of typical transfer cells. Therefore, the differentiation of the transfer cell-like structures, accompanied by the high-level expression of ZHA1, in the epidermal cells of mature leaves in particular may be important for the excretion of salt by these cells. PMID:8587992

  9. Albumin solder covalently bound to a polymer membrane: New approach to improve binding strength in laser tissue soldering in-vitro.

    Science.gov (United States)

    Hiebl, B; Ascher, L; Luetzow, K; Kratz, K; Gruber, C; Mrowietz, C; Nehring, M E; Lendlein, A; Franke, R-P; Jung, F

    2018-01-01

    Laser tissue soldering (LTS) based on indocyanine green (ICG)-mediated heat-denaturation of proteins might be a promising alternative technique for micro-suturing, but up to now the problem of too weak shear strength of the solder welds in comparison to sutures is not solved. Earlier reports gave promising results showing that solder supported by carrier materials can enhance the cohesive strength of the liquid solder. In these studies, the solder was applied to the carriers by dip coating. Higher reliability of the connection between the solder and the carrier material is expected when the solder is bound covalently to the carrier material. In the present study a poly(ether imide) (PEI) membrane served as carrier material and ICG-supplemented albumin as solder substrate. The latter was covalently coupled to the carrier membrane under physiological conditions to prevent structural protein changes. As laser source a diode continuous-wave laser emitting at 808 nm with intensities between 250 mW and 1500 mW was utilized. The albumin functionalized carrier membrane was placed onto the tunica media of explanted pig thoracic aortae forming an overlapping area of approximately 0.5×0.5 cm2. All tests were performed in a dry state to prevent laser light absorption by water. Infrared spectroscopy, spectro-photometrical determination of the secondary and primary amine groups after acid orange II staining, contact angle measurements, and atomic force microscopy proved the successful functionalization of the PEI membrane with albumin. A laser power of 450 mW LTS could generate a membrane-blood vessel connection which was characterized by a shear strength of 0.08±0.002 MPa, corresponding to 15% of the tensile strength of the native blood vessel. Theoretically, an overlapping zone of 4.1 mm around the entire circumference of the blood vessel could have provided shear strength of the PEI membrane-blood vessel compound identical to the tensile strength of the native

  10. Premature rupture of membranes

    Science.gov (United States)

    ... gov/ency/patientinstructions/000512.htm Premature rupture of membranes To use the sharing features on this page, ... water that surrounds your baby in the womb. Membranes or layers of tissue hold in this fluid. ...

  11. Higher levels of ethyl paraben and butyl paraben in rat amniotic fluid than in maternal plasma after subcutaneous administration

    DEFF Research Database (Denmark)

    Frederiksen, Hanne; Taxvig, Camilla; Hass, Ulla

    2008-01-01

    to obtain more knowledge about the distribution of ethyl paraben and butyl paraben in pregnant rats and pups after perinatal exposure, the presented study was designed. The data show response and distribution of ethyl paraben and butyl paraben in maternal rat plasma, pools of amniotic fluids, placenta......, whole-body fetuses, and in fetal liver after dosing of dams with 100, 200, and 400 mg/kg body weight (bw)/day from gestational day 7 to 21. After cesarean section of dams, the fluids and tissues were collected, deconjugated, and purified by solid-phase extraction, and ethyl paraben and butyl paraben...... were analyzed by liquid chromatography-tandem mass spectrometry. Markedly higher levels of ethyl paraben compared to butyl paraben were found in all fluids and tissues. Both ethyl paraben and butyl paraben in maternal plasma, livers, and whole-body tissues from fetus seemed to be saturated after dosing...

  12. Regenerative and Antibacterial Properties of Acellular Fish Skin Grafts and Human Amnion/Chorion Membrane: Implications for Tissue Preservation in Combat Casualty Care.

    Science.gov (United States)

    Magnusson, Skuli; Baldursson, Baldur Tumi; Kjartansson, Hilmar; Rolfsson, Ottar; Sigurjonsson, Gudmundur Fertram

    2017-03-01

    Improvised explosive devices and new directed energy weapons are changing warfare injuries from penetrating wounds to large surface area thermal and blast injuries. Acellular fish skin is used for tissue repair and during manufacturing subjected to gentle processing compared to biologic materials derived from mammals. This is due to the absence of viral and prion disease transmission risk, preserving natural structure and composition of the fish skin graft. The aim of this study was to assess properties of acellular fish skin relevant for severe battlefield injuries and to compare those properties with those of dehydrated human amnion/chorion membrane. We evaluated cell ingrowth capabilities of the biological materials with microscopy techniques. Bacterial barrier properties were tested with a 2-chamber model. The microstructure of the acellular fish skin is highly porous, whereas the microstructure of dehydrated human amnion/chorion membrane is mostly nonporous. The fish skin grafts show superior ability to support 3-dimensional ingrowth of cells compared to dehydrated human amnion/chorion membrane (p fish skin is a bacterial barrier for 24 to 48 hours. The unique biomechanical properties of the acellular fish skin graft make it ideal to be used as a conformal cover for severe trauma and burn wounds in the battlefield. Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.

  13. Computational modeling of adherent cell growth in a hollow-fiber membrane bioreactor for large-scale 3-D bone tissue engineering.

    Science.gov (United States)

    Mohebbi-Kalhori, Davod; Behzadmehr, Amin; Doillon, Charles J; Hadjizadeh, Afra

    2012-09-01

    The use of hollow-fiber membrane bioreactors (HFMBs) has been proposed for three-dimensional bone tissue growth at the clinical scale. However, to achieve an efficient HFMB design, the relationship between cell growth and environmental conditions must be determined. Therefore, in this work, a dynamic double-porous media model was developed to determine nutrient-dependent cell growth for bone tissue formation in a HFMB. The whole hollow-fiber scaffold within the bioreactor was treated as a porous domain in this model. The domain consisted of two interpenetrating porous regions, including a porous lumen region available for fluid flow and a porous extracapillary space filled with a collagen gel that contained adherent cells for promoting long-term growth into tissue-like mass. The governing equations were solved numerically and the model was validated using previously published experimental results. The contributions of several bioreactor design and process parameters to the performance of the bioreactor were studied. The results demonstrated that the process and design parameters of the HFMB significantly affect nutrient transport and thus cell behavior over a long period of culture. The approach presented here can be applied to any cell type and used to develop tissue engineering hollow-fiber scaffolds.

  14. In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration

    Directory of Open Access Journals (Sweden)

    T.I. Wodewotzky

    2012-12-01

    Full Text Available Support structures for dermal regeneration are composed of biodegradable and bioresorbable polymers, animal skin or tendons, or are bacteria products. The use of such materials is controversial due to their low efficiency. An important area within tissue engineering is the application of multipotent mesenchymal stromal cells (MSCs to reparative surgery. The combined use of biodegradable membranes with stem cell therapy may lead to promising results for patients undergoing unsuccessful conventional treatments. Thus, the aim of this study was to test the efficacy of using membranes composed of anionic collagen with or without the addition of hyaluronic acid (HA as a substrate for adhesion and in vitro differentiation of bone marrow-derived canine MSCs. The benefit of basic fibroblast growth factor (bFGF on the differentiation of cells in culture was also tested. MSCs were collected from dog bone marrow, isolated and grown on collagen scaffolds with or without HA. Cell viability, proliferation rate, and cellular toxicity were analyzed after 7 days. The cultured cells showed uniform growth and morphological characteristics of undifferentiated MSCs, which demonstrated that MSCs successfully adapted to the culture conditions established by collagen scaffolds with or without HA. This demonstrates that such scaffolds are promising for applications to tissue regeneration. bFGF significantly increased the proliferative rate of MSCs by 63% when compared to groups without the addition of the growth factor. However, the addition of bFGF becomes limiting, since it has an inhibitory effect at high concentrations in culture medium.

  15. In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration

    International Nuclear Information System (INIS)

    Wodewotzky, T.I.; Lima-Neto, J.F.; Pereira-Júnior, O.C.M.; Sudano, M.J.; Lima, S.A.F.; Bersano, P.R.O.; Yoshioka, S.A.; Landim-Alvarenga, F.C.

    2012-01-01

    Support structures for dermal regeneration are composed of biodegradable and bioresorbable polymers, animal skin or tendons, or are bacteria products. The use of such materials is controversial due to their low efficiency. An important area within tissue engineering is the application of multipotent mesenchymal stromal cells (MSCs) to reparative surgery. The combined use of biodegradable membranes with stem cell therapy may lead to promising results for patients undergoing unsuccessful conventional treatments. Thus, the aim of this study was to test the efficacy of using membranes composed of anionic collagen with or without the addition of hyaluronic acid (HA) as a substrate for adhesion and in vitro differentiation of bone marrow-derived canine MSCs. The benefit of basic fibroblast growth factor (bFGF) on the differentiation of cells in culture was also tested. MSCs were collected from dog bone marrow, isolated and grown on collagen scaffolds with or without HA. Cell viability, proliferation rate, and cellular toxicity were analyzed after 7 days. The cultured cells showed uniform growth and morphological characteristics of undifferentiated MSCs, which demonstrated that MSCs successfully adapted to the culture conditions established by collagen scaffolds with or without HA. This demonstrates that such scaffolds are promising for applications to tissue regeneration. bFGF significantly increased the proliferative rate of MSCs by 63% when compared to groups without the addition of the growth factor. However, the addition of bFGF becomes limiting, since it has an inhibitory effect at high concentrations in culture medium

  16. In vitro cultivation of canine multipotent mesenchymal stromal cells on collagen membranes treated with hyaluronic acid for cell therapy and tissue regeneration

    Energy Technology Data Exchange (ETDEWEB)

    Wodewotzky, T.I.; Lima-Neto, J.F. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Pereira-Júnior, O.C.M. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Departamento de Cirurgia e Anestesiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Sudano, M.J.; Lima, S.A.F. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Bersano, P.R.O. [Departamento de Patologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil); Yoshioka, S.A. [Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos, SP (Brazil); Landim-Alvarenga, F.C. [Departamento de Reprodução Animal e Radiologia Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual de São Paulo, Botucatu, SP (Brazil)

    2012-09-21

    Support structures for dermal regeneration are composed of biodegradable and bioresorbable polymers, animal skin or tendons, or are bacteria products. The use of such materials is controversial due to their low efficiency. An important area within tissue engineering is the application of multipotent mesenchymal stromal cells (MSCs) to reparative surgery. The combined use of biodegradable membranes with stem cell therapy may lead to promising results for patients undergoing unsuccessful conventional treatments. Thus, the aim of this study was to test the efficacy of using membranes composed of anionic collagen with or without the addition of hyaluronic acid (HA) as a substrate for adhesion and in vitro differentiation of bone marrow-derived canine MSCs. The benefit of basic fibroblast growth factor (bFGF) on the differentiation of cells in culture was also tested. MSCs were collected from dog bone marrow, isolated and grown on collagen scaffolds with or without HA. Cell viability, proliferation rate, and cellular toxicity were analyzed after 7 days. The cultured cells showed uniform growth and morphological characteristics of undifferentiated MSCs, which demonstrated that MSCs successfully adapted to the culture conditions established by collagen scaffolds with or without HA. This demonstrates that such scaffolds are promising for applications to tissue regeneration. bFGF significantly increased the proliferative rate of MSCs by 63% when compared to groups without the addition of the growth factor. However, the addition of bFGF becomes limiting, since it has an inhibitory effect at high concentrations in culture medium.

  17. Bioprinted Amniotic Fluid-Derived Stem Cells Accelerate Healing of Large Skin Wounds

    Science.gov (United States)

    Skardal, Aleksander; Mack, David; Kapetanovic, Edi; Atala, Anthony; Jackson, John D.; Yoo, James

    2012-01-01

    Stem cells obtained from amniotic fluid show high proliferative capacity in culture and multilineage differentiation potential. Because of the lack of significant immunogenicity and the ability of the amniotic fluid-derived stem (AFS) cells to modulate the inflammatory response, we investigated whether they could augment wound healing in a mouse model of skin regeneration. We used bioprinting technology to treat full-thickness skin wounds in nu/nu mice. AFS cells and bone marrow-derived mesenchymal stem cells (MSCs) were resuspended in fibrin-collagen gel and “printed” over the wound site. At days 0, 7, and 14, AFS cell- and MSC-driven wound closure and re-epithelialization were significantly greater than closure and re-epithelialization in wounds treated by fibrin-collagen gel only. Histological examination showed increased microvessel density and capillary diameters in the AFS cell-treated wounds compared with the MSC-treated wounds, whereas the skin treated only with gel showed the lowest amount of microvessels. However, tracking of fluorescently labeled AFS cells and MSCs revealed that the cells remained transiently and did not permanently integrate in the tissue. These observations suggest that the increased wound closure rates and angiogenesis may be due to delivery of secreted trophic factors, rather than direct cell-cell interactions. Accordingly, we performed proteomic analysis, which showed that AFS cells secreted a number of growth factors at concentrations higher than those of MSCs. In parallel, we showed that AFS cell-conditioned media induced endothelial cell migration in vitro. Taken together, our results indicate that bioprinting AFS cells could be an effective treatment for large-scale wounds and burns. PMID:23197691

  18. Tissue expression and enzymologic characterization of human prostate specific membrane antigen and its rat and pig orthologs

    Czech Academy of Sciences Publication Activity Database

    Rovenská, Miroslava; Hlouchová, Klára; Šácha, Pavel; Mlčochová, Petra; Horák, Vratislav; Zámečník, J.; Bařinka, C.; Konvalinka, Jan

    2008-01-01

    Roč. 68, č. 2 (2008), s. 171-182 ISSN 0270-4137 R&D Projects: GA MŠk 1M0508; GA ČR GA524/04/0102 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50450515 Keywords : prostate specific membrane antigen * glutamate carboxypeptidase II * animal orthologs * prostate cancer * animal model Subject RIV: CE - Biochemistry Impact factor: 3.069, year: 2008

  19. A randomized controlled trial of foley catheter, extra-amniotic saline infusion and prostaglandin e2 suppository for labor induction.

    Directory of Open Access Journals (Sweden)

    Mandana Mansour Ghanaie

    2013-06-01

    Full Text Available The aim of this study is to further compare the efficacy of PGE2 suppository, the intracervical foley catheter and extra-amniotic saline infusion in nulliparous women referred for labor induction.Totally 368 nulliparous women with a Bishop score ≤ 4 with singleton gestation, vertex presentation and intact membrane referred for labor induction were randomly assigned to 3 groups; Foley catheter alone, Extra-amniotic saline infusion (EASI and PGE2 suppository. All women received concurrent dilute oxytocine infusion. The change in the Bishop Score, labor progress, various labor endpoints and outcomes of labor were assessed.From 363 women studied after exclusion of 5, 119 were assigned to EASI, 121 to Foley and 118 to PGE2. Patients' demographics did not differ significantly between three groups nor did indication for induction (P = 0.0001. The EASI group had a significant improvement in Bishop Score 6 hours after induction. The mean time to active phase was 357±135min for EASI,457±178 for Foley and 609±238 min for PGE2 group respectively (P < 0.05.rate of spontaneous rupture of membranes was higher in the EASI group (P = 0.0001 and the mean time from the start of induction up to spontaneous rupture of membranes in the EASI group was shorter than other group(P < 0.05. The mean time to vaginal delivery was 14.8±6.1 in EASI group,11.4±4.8 in Foley and 18.9±6.4 in PGE2 group(P < 0.05.there were no differences in Apgar scores, mean neonatal birth weight and neonatal morbidity.Our study showed that pre-induction cervical ripening by EASI with concurrent oxytocin is better than Foley and PGE2 in Bishop score and various labor end point and outcomes.

  20. Amniotic oxytocin and vasopressin in relation to human fetal development and labour

    NARCIS (Netherlands)

    Oosterbaan, H. P.; Swaab, D. F.

    1989-01-01

    Previous experiments in rats revealed increased amniotic oxytocin (OXT) levels in the course of normal development and increased vasopressin (AVP) levels in retarded fetal growth. In order to see whether similar changes would also occur in human, OXT and AVP levels were determined in amniotic fluid,

  1. Processing laboratory of radio sterilized biological tissues

    International Nuclear Information System (INIS)

    Aguirre H, Paulina; Zarate S, Herman; Silva R, Samy; Hitschfeld, Mario

    2005-01-01

    The nuclear development applications have also reached those areas related to health. The risk of getting contagious illnesses through applying biological tissues has been one of the paramount worries to be solved since infectious illnesses might be provoked by virus, fungis or bacterias coming from donors or whether they have been introduced by means of intermediate stages before the use of these tissues. Therefore it has been concluded that the tissue allografts must be sterilized. The sterilization of medical products has been one of the main applications of the ionizing radiations and that it is why the International Organization of Atomic Energy began in the 70s promoting works related to the biological tissue sterilization and pharmaceutical products. The development of different tissue preservation methods has made possible the creation of tissue banks in different countries, to deal with long-term preservation. In our country, a project was launched in 1998, 'Establishment of a Tissue Bank in Latino america', this project was supported by the OIEA through the project INT/ 6/ 049, and was the starting of the actual Processing Laboratory of Radioesterilized Biological Tissues (LPTR), leaded by the Chilean Nuclear Energy Commission (CCHEN). This first organization is part of a number of entities compounding the Tissue Bank in Chile, organizations such as the Transplantation Promotion Corporation hospitals and the LPTR. The working system is carried out by means of the interaction between the hospitals and the laboratory. The medical professionals perform the procuring of tissues in the hospitals, then send them to the LPTR where they are processed and sterilized with ionizing radiation. The cycle ends up with the tissues return released to the hospitals, where they are used, and then the result information is sent to the LPTR as a form of feedback. Up to now, human skin has been processed (64 donors), amniotic membranes (35 donors) and pig skin (175 portions

  2. Membrane-Tethered Delta-Like 1 Homolog (DLK1) Restricts Adipose Tissue Size By Inhibiting Preadipocyte Proliferation

    DEFF Research Database (Denmark)

    Mortensen, Sussi B; Jensen, Charlotte H; Schneider, Mikael

    2012-01-01

    Adipocyte renewal from preadipocytes has been shown to occur throughout life and to contribute to obesity, yet very little is known about the molecular circuits that control preadipocyte expansion. The soluble form of the preadipocyte factor (also known as pref-1) delta-like 1 homolog (DLK1(S...... in vivo. Here, we demonstrate for the first time that only membrane-bound DLK1 (DLK1(M)) exhibits a substantial repression effect on preadipocyte proliferation. Thus, by independently manipulating DLK1 isoform levels, we established that DLK1(M) inhibits G1-to-S-phase cell cycle progression and thereby...

  3. Intraamniotic Inflammation in Women with Preterm Prelabor Rupture of Membranes.

    Directory of Open Access Journals (Sweden)

    Ivana Musilova

    Full Text Available To characterize subgroups of preterm prelabor rupture of membranes (PPROM and short-term neonatal outcomes based on the presence and absence of intraamniotic inflammation (IAI and/or microbial invasion of the amniotic cavity (MIAC.One hundred and sixty-six Caucasian women with singleton pregnancies were included in this study. Amniotic fluid samples were obtained by transabdominal amniocentesis (n=166 and were assayed for interleukin-6 levels by a lateral flow immunoassay. The presence of Ureaplasma species, Mycoplasma hominis, Chlamydia trachomatis, and 16S rRNA was evaluated in the amniotic fluid. IAI was defined as amniotic fluid IL-6 values, measured by a point of care test, higher than 745 pg/mL.Microbial-associated IAI (IAI with MIAC and sterile intraamniotic inflammation (IAI alone were found in 21% and 4%, respectively, of women with PPROM. Women with microbial-associated IAI had higher microbial loads of Ureaplasma species in the amniotic fluid than women with MIAC alone. No differences in the short-term neonatal morbidity with respect to the presence of microbial-associated IAI, sterile IAI and MIAC alone were found after adjusting for the gestational age at delivery in women with PPROM.Microbial-associated but not sterile intraamniotic inflammation is common in Caucasian women with PPROM. The gestational age at delivery but not the presence of inflammation affects the short-term neonatal morbidity of newborns from PPROM pregnancies.

  4. Prostate specific membrane antigen (PSMA) is a tissue-specific target for adenoviral transduction of prostate cancer in vitro

    NARCIS (Netherlands)

    Kraaij, R; van Rijswijk, ALCT; Haisma, HJ; Bangma, CH

    2005-01-01

    BACKGROUND. Adenovirus binds to the coxsackievirus and adenovirus receptor (CAR) as a first step in the process of cellular infection. This dependence on CAR potentially limits the use of adenovirus in gene therapy, since CAR is expressed in many tissues of the body, and, expression of CAR may be

  5. Effects of maternal subclinical hypothyroidism on amniotic fluid cells oxidative status.

    Science.gov (United States)

    Novakovic, Tanja R; Dolicanin, Zana C; Djordjevic, Natasa Z

    2018-06-01

    In this study, we researched the effects of maternal subclinical hypothyroidism on the amniotic fluid cells oxidative metabolism during the first trimester of pregnancy. Oxidative stress and damage biomarkers were assayed in the amniotic fluid cells of healthy and pregnant women with subclinical hypothyroidism. Obtained results show that amniotic fluid cells of pregnant women with subclinical hypothyroidism have significantly higher concentrations of oxidative stress biomarkers (superoxide anion, nitric oxide, peroxynitrite) and oxidative damage (lipid peroxide and micronuclei frequency), but lower concentrations of hydrogen peroxide and oxidized glutathione in comparison to healthy pregnant women. We also showed that oxidative stress biomarkers were positively correlated with micronuclei frequency and lipid peroxide concentration in amniotic fluid cells of pregnant women with subclinical hypothyroidism. The present study provides the first evidence for prooxidative effects of maternal subclinical hypothyroidism on the fetus obtained by the estimating oxidative metabolism in the amniotic fluid cells. Copyright © 2018 Elsevier Inc. All rights reserved.

  6. [PELVIS/SACRAL syndrome with livedoid haemangioma and amniotic band].

    Science.gov (United States)

    Bourrat, E; Lemarchand-Venencie, F; Jacquemont, M-L; El Ghoneimi, A; Wassef, M; Leger, J; Morel, P

    2008-12-01

    PELVIS or SACRAL syndrome denotes the association of local haemangioma and malformation in the pelvic region. In this paper, we report a case noteworthy on account of the initially livedoid appearance of the haemangioma as well as associated amniotic banding of an upper limb. A newborn male infant underwent left colostomy on the day of birth due to anal imperforation and anomalies of the external genital organs with sexual ambiguity. Examination of the skin and appendages revealed poorly delineated hypopigmentation in the sacrolumbar region and a fibrous groove around the right arm characteristic of amniotic band syndrome. Sacrolumbar and pelvic MRI scans revealed deviation towards the left of the last three sacral vertebrae with no medullary anomalies. Retrograde cystography showed a recto-uretral fistula. Progression of the infant's condition was marked by the appearance during the first month of a flat, violaceous, angiomatous, livedoid lesion in the middle of the buttocks and the perineum and a linear lesion on the rear aspect of the right lower limb. The skin biopsy of this lesion revealed a single capillary lobule at the dermal-hypodermal junction of non-specific appearance but with marked Glut1 expression by endothelial cells highly evocative of infantile haemangioma. Segmented haemangiomas are commonly associated with extracutaneous abnormalities. By analogy with PHACE syndrome, defined as association of segmented facial haemangioma with cerebral, ocular and cardio-aortic abnormalities, PELVIS/SACRAL syndrome denotes the association of segmented haemangioma of the loins (sacrolumbar region, buttocks or perineum=napkin haemangioma) with spinal dysraphia affecting the sacrolumbar spine, the terminal medullary cone, the genitourinary organs and the anal region to different degrees. Diagnosis of haemangioma associated with PELVIS/SACRAL syndrome may be delayed or complicated due to the macular, telangiectasic or livedoid appearance commonly seen. To our

  7. (3H)leukotriene B4 binding to the guinea pig spleen membranes: a rich tissue source for a high affinity leukotriene B4 receptor site

    International Nuclear Information System (INIS)

    Cheng, J.B.; Kohi, F.; Townley, R.G.

    1986-01-01

    To select a tissue rich for the high affinity leukotriene (LT)B 4 receptor site, they compared binding of 1 nM ( 3 H)LTB 4 (180 Ci/mmol) to the crude membrane preparations of guinea pig spleen, thymus, lung, uterus, bladder, brain, adrenal gland, small intestine, liver, kidney and heart. They found that the membrane preparations from spleen contained the highest binding activity per mg protein. They characterized the LTB 4 binding to the spleen preparation in detail. LTB 4 binding was rapid, reversible, stereoselective and saturable. The data from equilibrium experiments showed a linear Scatchard plot with a K/sub d/ of 1.6 nM and a binding site density of 259 fmol/mg prot. The rank order of agents competing for spleen ( 3 H)LTB 4 binding at 25 0 C was: LTB 4 (K/sub i/ = 2.8 nM) > 20-OH-LTB 4 (23 nM) > LTA 4 (48 nM) > LTA 4 methyl ester (0.13 μM) > 20-COOH-LTB 4 (> 6.6 μM) ≥ arachidonic acid (0.15 mM) similarly ordered FPL-55,712 (0.11 mM). At 4 0 C, LTB 4 (2.3 nM) competed at least 10x more effectively than 20-OH-LTB 4 (29 nM) and 20-COOH-LTB 4 (> 6.6 μM). HPLC analysis indicated that incubation of 84 ng LTB 4 with the spleen membrane at 25 0 C did not result in the formation of 20-OH-LTB 4 ( 3 H)LTB 4 receptor binding sites

  8. Membranous glomerulonephritis in a patient with anti-u1 ribonucleoprotein (RNP antibody-positive mixed connective tissue disease: A case report

    Directory of Open Access Journals (Sweden)

    Naoya Toriu

    2018-03-01

    Full Text Available We report a 33-year-old Japanese man diagnosed with mixed connective tissue disease (MCTD who developed nephrotic proteinuria. Both speckled antinuclear antibody (ANA and anti-U1 ribonucleoprotein (RNP antibody were positive, but anti-double-stranded DNA (dsDNA antibody and anti-Smith (Sm antibody were negative, while complement levels were normal. Renal biopsy revealed membranous glomerulonephritis (MGN with diffuse thickening of the glomerular basement membrane (GBM plus spike and bubble formation. Immunofluorescence demonstrated granular deposits of IgG and C3 along the GBM. Analysis of IgG subclasses showed predominant deposition of IgG1 and IgG4, unlike typical lupus nephritis in which there is predominant deposition of IgG1, IgG2, IgG3, and C1q. Electron microscopy identified numerous large electron-dense deposits (EDD of various types in the subepithelial region of the GBM, but there were no EDD localized in the mesangium or subendothelium. Based on these findings, MGN was considered to be closely related to MCTD in this patient.

  9. Ultrastructural and cytochemical study of membrane alterations in x-irradiated liver tissue from normal and vitamin E-deficient ducklings

    International Nuclear Information System (INIS)

    Huijbers, W.A.R.; Oosterbaan, J.A.; Meskendorp-Haarsma, T.J.; Hardonk, M.J.; Molenaar, I.

    1979-01-01

    An investigation into the differential susceptibility of liver cellular membranes to peroxidative processes has been performed, using x irradiation on the liver surface, resulting in a a 3-mm penetrating gradient of membrane damage. Ultrastructural, cytochemical, and histochemical findings in this area point to a differential sensitivity of cellular membranes to x irradiation. The plasma membrane and the lysosomal membrane, containing much lipid and cholesterol and little membrane and the lysosomal membrane, containing much lipid and cholesterol and little vitamin E, are highly susceptible to x irradiation. Less sensitive are the membranes of mitochondria and endoplasmic reticulum, containing relatively much vitamin E and proteins and a lower amount of lipids and cholesterol

  10. Regulation of osteogenesis of human amniotic mesenchymal stem cells by sodium butyrate.

    Science.gov (United States)

    Fan, Xiaoting; Li, Lei; Ye, Zhaoyang; Zhou, Yan; Tan, Wen-Song

    2018-04-01

    Human amniotic membrane-derived mesenchymal stem cells (hAMSCs) draw great interests for regenerative medicine due to convenient availability and low immunogenicity. However, suboptimal culture conditions limit their application. In recent years, small molecules have proven powerful in regulating stem cell fates and can be applied to stimulate their function. In the present study, the impacts of sodium butyrate (NaBu), a histone deacetylase inhibitor (HDACi), on hAMSCs were investigated. It was shown that NaBu at a low concentration inhibited cell proliferation by arresting cell cycle at G0/G1 rather than inducing apoptosis. When NaBu was supplemented at a concentration of generated and the expression of osteogenesis-related genes (ALP, Runx2, Opn, and Ocn) and proteins (Col1a1, OPN, OCN, Runx2, and TAZ) were both significantly enhanced. However, a higher concentration (1.0 mM) and longer exposure time (14 days) of NaBu showed no such effects, which may be partially attributed to both the increased expression of histone deacetylase 8 (HDAC8) and reduced level of H3K9-Ace, thus leading to the transcriptional inhibition during osteogenesis. Further, it was indicated that ERK might be involved in the stimulatory effects of NaBu. These findings may be helpful to develop an efficient culture process for hAMSCs towards bone regeneration. © 2018 International Federation for Cell Biology.

  11. Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication

    Directory of Open Access Journals (Sweden)

    Natascha Krömmelbein

    2016-02-01

    Full Text Available The human cytomegalovirus (HCMV replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming functions of adenoviruses have been notoriously resistant to HCMV replication and progeny production. Here, we provide first-time evidence that a permanent cell line immortalized by adenovirus type 5 E1A and E1B (CAP is supporting the full HCMV replication cycle and is releasing infectious progeny. The CAP cell line had previously been established from amniotic fluid cells which were likely derived from membranes of the developing fetus. These cells can be grown under serum-free conditions. HCMV efficiently penetrated CAP cells, expressed its immediate-early proteins and dispersed restrictive PML-bodies. Viral DNA replication was initiated and viral progeny became detectable by electron microscopy in CAP cells. Furthermore, infectious virus was released from CAP cells, yet to lower levels compared to fibroblasts. Subviral dense bodies were also secreted from CAP cells. The results show that E1A/E1B expression in transformed cells is not generally repressive to HCMV replication and that CAP cells may be a good substrate for dense body based vaccine production.

  12. The Implication of PGC-1α on Fatty Acid Transport across Plasma and Mitochondrial Membranes in the Insulin Sensitive Tissues

    Directory of Open Access Journals (Sweden)

    Elżbieta Supruniuk

    2017-11-01

    Full Text Available PGC-1α coactivator plays a decisive role in the maintenance of lipid balance via engagement in numerous metabolic processes (i.e., Krebs cycle, β-oxidation, oxidative phosphorylation and electron transport chain. It constitutes a link between fatty acids import and their complete oxidation or conversion into bioactive fractions through the coordination of both the expression and subcellular relocation of the proteins involved in fatty acid transmembrane movement. Studies on cell lines and/or animal models highlighted the existence of an upregulation of the total and mitochondrial FAT/CD36, FABPpm and FATPs content in skeletal muscle in response to PGC-1α stimulation. On the other hand, the association between PGC-1α level or activity and the fatty acids transport in the heart and adipocytes is still elusive. So far, the effects of PGC-1α on the total and sarcolemmal expression of FAT/CD36, FATP1, and FABPpm in cardiomyocytes have been shown to vary in relation to the type of PPAR that was coactivated. In brown adipose tissue (BAT PGC-1α knockdown was linked with a decreased level of lipid metabolizing enzymes and fatty acid transporters (FAT/CD36, FABP3, whereas the results obtained for white adipose tissue (WAT remain contradictory. Furthermore, dysregulation in lipid turnover is often associated with insulin intolerance, which suggests the coactivator's potential role as a therapeutic target.

  13. Meconium concentration and amniotic fluid index influence the outcome of amnioinfusion.

    Science.gov (United States)

    Puertas, A; Carrillo, M P; çlvarez, M; Cañizares, J M; Miño, M; Malde, J

    2001-10-01

    To determine the usefulness of amnioinfusion as a function of meconium concentration and amniotic fluid index. This was a prospective study of 206 pregnant women in whom amniotic fluid was moderately or heavily stained with meconium, according to subjective evaluation. The women were assigned randomly to receive amnioinfusion (n=103) or no amnioinfusion (control group, n=103). The results were compared in women with =/15 % meconium in the amniotic fluid (measured by centrifugation), and in women in whom the amniotic fluid index calculated 60 min after insertion of the amnioinfusion catheter was 10. In women with >15% meconium, amnioinfusion decreased the rate of cesarian sections motivated by fetal distress (2.5% vs 22.2%), and in women with =/amnioinfusion decreased the presence of meconium below the vocal cords (6.4% vs 25.9%). Greater benefits after amnioinfusion were seen in women with an amniotic fluid index =/>10: the rate of cesarian sections was lower (1.3% vs 13.3%), as was the frequency of meconium below the vocal cords (10.1% vs 33.3%). Beneficial effects of amnioinfusion were seen in women with high and low concentrations of meconium, and with high and low amniotic fluid indexes. These criteria should therefore not be used to decide whether amnioinfusion is indicated when the amniotic fluid is moderately or heavily stained with meconium.

  14. The Immunomodulatory Properties of Amniotic Cells: The Two Sides of the Coin.

    Science.gov (United States)

    Magatti, Marta; Vertua, Elsa; Cargnoni, Anna; Silini, Antonietta; Parolini, Ornella

    2018-01-01

    Among the many cell types useful in developing therapeutic treatments, human amniotic cells from placenta have been proposed as valid candidates. Both human amniotic epithelial and mesenchymal stromal cells, and the conditioned medium generated from their culture, exert multiple immunosuppressive activities. Indeed, they inhibit T and B cell proliferation, suppress inflammatory properties of monocytes, macrophages, dendritic cells, neutrophils, and natural killer cells, while promoting induction of cells with regulatory functions such as regulatory T cells and anti-inflammatory M2 macrophages. These properties have laid the foundation for their use for the treatment of inflammatory-based diseases, and encouraging results have been obtained in different preclinical disease models where exacerbated inflammation is present. Moreover, an immune-privileged status of amniotic cells has been often highlighted. However, even if long-term engraftment of amniotic cells has been reported into immunocompetent animals, only few cells survive after infusion. Furthermore, amniotic cells have been shown to be able to induce immune responses in vivo and, under specific culture conditions, they can stimulate T cell proliferation in vitro. Although immunosuppressive properties are a widely recognized characteristic of amniotic cells, immunogenic and stimulatory activities appear to be less reported, sporadic events. In order to improve therapeutic outcome, the mechanisms responsible for the suppressive versus stimulatory activity need to be carefully addressed. In this review, both the immunosuppressive and immunostimulatory activity of amniotic cells will be discussed.

  15. Anxiolytic-Like Actions of Fatty Acids Identified in Human Amniotic Fluid

    Directory of Open Access Journals (Sweden)

    Rosa Isela García-Ríos

    2013-01-01

    Full Text Available Eight fatty acids (C12–C18 were previously identified in human amniotic fluid, colostrum, and milk in similar proportions but different amounts. Amniotic fluid is well known to be the natural environment for development in mammals. Interestingly, amniotic fluid and an artificial mixture of fatty acids contained in amniotic fluid produce similar anxiolytic-like actions in Wistar rats. We explored whether the lowest amount of fatty acids contained in amniotic fluid with respect to colostrum and milk produces such anxiolytic-like effects. Although a trend toward a dose-response effect was observed, only an amount of fatty acids that was similar to amniotic fluid fully mimicked the effect of diazepam (2 mg/kg, i.p. in the defensive burying test, an action devoid of effects on locomotor activity and motor coordination. Our results confirm that the amount of fatty acids contained in amniotic fluid is sufficient to produce anxiolytic-like effects, suggesting similar actions during intrauterine development.

  16. Oxidative stress biomarkers in amniotic fluid of pregnant women with hypothyroidism.

    Science.gov (United States)

    Novakovic, Tanja R; Dolicanin, Zana C; Djordjevic, Natasa Z

    2017-11-15

    Hypothyroidism in pregnancy is the serious state that may lead to fetal morbidity and mortality. Oxidative stress biomarkers in the amniotic fluid can provide important information on the health, development and maturation of the fetus during pregnancy. In this study, we examined whether maternal hypothyroidism contributes to increased oxidative stress biomarkers in the amniotic fluid during the first trimester of pregnancy. The study was conducted on healthy pregnant women and pregnant women with hypothyroidism (gestational age: 16-18 weeks). Oxidative stress biomarkers, such as superoxide anion (O 2 •- ), hydrogen peroxide (H 2 O 2 ), nitric oxide (NO), peroxynitrite (ONOO - ), lipid peroxide (LPO), reduced glutathione (GSH) and oxidized glutathione (GSSG) were assayed in the amniotic fluid. The results of this study indicated that concentrations of O 2 •- and NO are significantly higher, while the concentration of H 2 O 2 is significantly lower in the amniotic fluid of pregnant women with hypothyroidism in comparison to healthy pregnant women. There were no differences in concentrations of LPO, GSH and GSSG among tested groups. Also, we found that amniotic fluid concentration of O 2 •- is negatively correlated with the body weight and Apgar score values of the newborns. These results suggest that pregnancy hypothyroidism is characterized by the amniotic fluid oxidative stress. Incorporation of the oxidative stress biomarkers measurement in the amniotic fluid may be of clinical importance in the management of pregnancy hypothyroidism.

  17. Milk Fat Globule Membrane Attenuates High-Fat Diet-Induced Obesity by Inhibiting Adipogenesis and Increasing Uncoupling Protein 1 Expression in White Adipose Tissue of Mice

    Directory of Open Access Journals (Sweden)

    Tiange Li

    2018-03-01

    Full Text Available Milk fat globule membrane (MFGM, a protein-lipid complex surrounding the fat globules in milk, has many health benefits. The aim of the current study was to investigate whether MFGM could prevent obesity through inhibiting adipogenesis and promoting brown remodeling of white adipose tissue (WAT in mice fed with high-fat diet. C57BL/6 mice were fed a normal diet (ND, high-fat diet (HFD, HFD plus MFGM at 100 mg/kg BW, 200 mg/kg BW or 400 mg/kg BW for 8 weeks. Results showed that MFGM suppressed body weight gain induced by HFD, reduced white adipose tissue (WAT mass accompanied with the decrease in adipocyte sizes. MFGM was found to have partially improved serum lipid profiles, as well as to have suppressed HFD-induced adipogenesis as shown by reduced expression of peroxisome proliferators-activator receptor-γ (PPARγ, CCAAT/enhancer-binding protein-α (C/EBPα and sterol regulatory element-binding protein-1c (SREBP-1c. MFGM also markedly increased the phosphorylation of AMP-activated protein kinase (AMPK and acetyl-CoA carboxylase (ACC, showing activation of AMPK pathway. Moreover, MFGM promoted browning of inguinal WAT by upregulation the protein expression of uncoupling protein 1 (UCP1 in HFD mice. Taken together, these findings provide evidence that MFGM may protect against diet-induced adiposity by suppressing adipogenesis and promoting brown-like transformation in WAT.

  18. A novel amniote model of epimorphic regeneration: the leopard gecko, Eublepharis macularius.

    Science.gov (United States)

    McLean, Katherine E; Vickaryous, Matthew K

    2011-08-16

    Epimorphic regeneration results in the restoration of lost tissues and structures from an aggregation of proliferating cells known as a blastema. Among amniotes the most striking example of epimorphic regeneration comes from tail regenerating lizards. Although tail regeneration is often studied in the context of ecological costs and benefits, details of the sequence of tissue-level events are lacking. Here we investigate the anatomical and histological events that characterize tail regeneration in the leopard gecko, Eublepharis macularius. Tail structure and tissue composition were examined at multiple days following tail loss, revealing a conserved pattern of regeneration. Removal of the tail results in a consistent series of morphological and histological events. Tail loss is followed by a latent period of wound healing with no visible signs of regenerative outgrowth. During this latent period basal cells of the epidermis proliferate and gradually cover the wound. An additional aggregation of proliferating cells accumulates adjacent to the distal tip of the severed spinal cord marking the first appearance of the blastema. Continued growth of the blastema is matched by the initiation of angiogenesis, followed by the re-development of peripheral axons and the ependymal tube of the spinal cord. Skeletal tissue differentiation, corresponding with the expression of Sox9, and muscle re-development are delayed until tail outgrowth is well underway. We demonstrate that tail regeneration in lizards involves a highly conserved sequence of events permitting the establishment of a staging table. We show that tail loss is followed by a latent period of scar-free healing of the wound site, and that regeneration is blastema-mediated. We conclude that the major events of epimorphic regeneration are highly conserved across vertebrates and that a comparative approach is an invaluable biomedical tool for ongoing regenerative research.

  19. High dietary level of synthetic vitamin E on lipid peroxidation, membrane fatty acid composition and cytotoxicity in breast cancer xenograft and in mouse host tissue

    Directory of Open Access Journals (Sweden)

    Barnes Christopher J

    2003-03-01

    Full Text Available Abstract Background d-α-tocopherol is a naturally occurring form of vitamin E not previously known to have antitumor activity. Synthetic vitamin E (sE is a commonly used dietary supplement consisting of a mixture of d-α-tocopherol and 7 equimolar stereoisomers. To test for antilipid peroxidation and for antitumor activity of sE supplementation, two groups of nude mice bearing a MDA-MB 231 human breast cancer tumor were fed an AIN-76 diet, one with and one without an additional 2000 IU/kg dry food (equivalent to 900 mg of all-rac-α-tocopherol or sE. This provided an intake of about 200 mg/kg body weight per day. The mice were killed at either 2 or 6 weeks after the start of dietary intervention. During necropsy, tumor and host tissues were excised for histology and for biochemical analyses. Results Tumor growth was significantly reduced by 6 weeks of sE supplementation. Thiobarbituric acid reactive substances, an indicator of lipid peroxidation, were suppressed in tumor and in host tissues in sE supplemented mice. In the sE treated mice, the fatty acid composition of microsomal and mitochondrial membranes of tumor and host tissues had proportionately less linoleic acid (n-6 C 18-2, similar levels of arachidonic acid (n-6 C 20-4, but more docosahexanoic acid (n-3 C 22-6. The sE supplementation had no significant effect on blood counts or on intestinal histology but gave some evidence of cardiac toxicity as judged by myocyte vacuoles and by an indicator of oxidative stress (increased ratio of Mn SOD mRNA over GPX1 mRNA. Conclusions At least one of the stereoisomers in sE has antitumor activity. Synthetic vitamin E appears to preferentially stabilize membrane fatty acids with more double bonds in the acyl chain. Although sE suppressed tumor growth and lipid peroxidation, it may have side-effects in the heart.

  20. Amniotic Mesenchymal Stromal Cells Exhibit Preferential Osteogenic and Chondrogenic Differentiation and Enhanced Matrix Production Compared With Adipose Mesenchymal Stromal Cells.

    Science.gov (United States)

    Topoluk, Natasha; Hawkins, Richard; Tokish, John; Mercuri, Jeremy

    2017-09-01

    Therapeutic efficacy of various mesenchymal stromal cell (MSC) types for orthopaedic applications is currently being investigated. While the concept of MSC therapy is well grounded in the basic science of healing and regeneration, little is known about individual MSC populations in terms of their propensity to promote the repair and/or regeneration of specific musculoskeletal tissues. Two promising MSC sources, adipose and amnion, have each demonstrated differentiation and extracellular matrix (ECM) production in the setting of musculoskeletal tissue regeneration. However, no study to date has directly compared the differentiation potential of these 2 MSC populations. To compare the ability of human adipose- and amnion-derived MSCs to undergo osteogenic and chondrogenic differentiation. Controlled laboratory study. MSC populations from the human term amnion were quantified and characterized via cell counting, histologic assessment, and flow cytometry. Differentiation of these cells in comparison to commercially purchased human adipose-derived mesenchymal stromal cells (hADSCs) in the presence and absence of differentiation media was evaluated via reverse transcription polymerase chain reaction (PCR) for bone and cartilage gene transcript markers and histology/immunohistochemistry to examine ECM production. Analysis of variance and paired t tests were performed to compare results across all cell groups investigated. The authors confirmed that the human term amnion contains 2 primary cell types demonstrating MSC characteristics-(1) human amniotic epithelial cells (hAECs) and (2) human amniotic mesenchymal stromal cells (hAMSCs)-and each exhibited more than 90% staining for MSC surface markers (CD90, CD105, CD73). Average viable hAEC and hAMSC yields at harvest were 2.3 × 10 6 ± 3.7 × 10 5 and 1.6 × 10 6 ± 4.7 × 10 5 per milliliter of amnion, respectively. As well, hAECs and hAMSCs demonstrated significantly greater osteocalcin ( P = .025), aggrecan ( P

  1. Eggshell membrane: A possible new natural therapeutic for joint and connective tissue disorders. Results from two open-label human clinical studies

    Directory of Open Access Journals (Sweden)

    Kevin J Ruff

    2009-05-01

    Full Text Available Kevin J Ruff1, Dale P DeVore2, Michael D Leu3, Mark A Robinson41ESM Technologies, LLC, Carthage, MO, USA; 2Membrell, LLC, Carthage, MO, USA; 3Private Practice, Jenks, OK, USA; 4Robinson Family Health Center, Carthage, MO, USABackground: Natural Eggshell Membrane (NEM® is a novel dietary supplement that contains naturally occurring glycosaminoglycans and proteins essential for maintaining healthy joint and connective tissues. Two single center, open-label human clinical studies were conducted to evaluate the efficacy and safety of NEM® as a treatment for pain and inflexibility associated with joint and connective tissue disorders. Methods: Eleven (single-arm trial and 28 (double-arm trial patients received oral NEM® 500 mg once daily for four weeks. The primary outcome measure was to evaluate the change in general pain associated with the treatment joints/areas (both studies. In the single-arm trial, range of motion (ROM and related ROM-associated pain was also evaluated. The primary treatment response endpoints were at seven and 30 days. Both clinical assessments were performed on the intent-to-treat (ITT population within each study.Results: Single-arm trial: Supplementation with NEM® produced a significant treatment response at seven days for flexibility (27.8% increase; P = 0.038 and at 30 days for general pain (72.5% reduction; P = 0.007, flexibility (43.7% increase; P = 0.006, and ROM-associated pain (75.9% reduction; P = 0.021. Double-arm trial: Supplementation with NEM® produced a significant treatment response for pain at seven days for both treatment arms (X: 18.4% reduction; P = 0.021. Y: 31.3% reduction; P = 0.014. There was no clinically meaningful difference between treatment arms at seven days, so the Y arm crossed over to the X formulation for the remainder of the study. The significant treatment response continued through 30 days for pain (30.2% reduction; P = 0.0001. There were no adverse events reported during either

  2. Cell cytoskeletal changes effected by static compressive stress lead to changes in the contractile properties of tissue regenerative collagen membranes

    Directory of Open Access Journals (Sweden)

    K Gellynck

    2013-06-01

    Full Text Available Static compressive stress can influence the matrix, which subsequently affects cell behaviour and the cell’s ability to further transform the matrix. This study aimed to assess response to static compressive stress at different stages of osteoblast differentiation and assess the cell cytoskeleton’s role as a conduit of matrix-derived stimuli. Mouse bone marrow mesenchymal stem cells (MSCs (D1 ORL UVA, osteoblastic cells (MC3T3-E1 and post-osteoblast/pre-osteocyte-like cells (MLO-A5 were seeded in hydrated and compressed collagen gels. Contraction was quantified macroscopically, and cell morphology, survival, differentiation and mineralisation assessed using confocal microscopy, alamarBlue® assay, real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR and histological stains, respectively. Confocal microscopy demonstrated cell shape changes and favourable microfilament organisation with static compressive stress of the collagen matrix; furthermore, cell survival was greater compared to the hydrated gels. The stage of osteoblast differentiation determined the degree of matrix contraction, with MSCs demonstrating the greatest amount. Introduction of microfilament disrupting inhibitors confirmed that pre-stress and tensegrity forces were under the influence of gel density, and there was increased survival and differentiation of the cells within the compressed collagen compared to the hydrated collagen. There was also relative stiffening and differentiation with time of the compressed cell-seeded collagen, allowing for greater manipulation. In conclusion, the combined collagen chemistry and increased density of the microenvironment can promote upregulation of osteogenic genes and mineralisation; MSCs can facilitate matrix contraction to form an engineered membrane with the potential to serve as a ‘pseudo-periosteum’ in the regeneration of bone defects.

  3. Pseudomonas aeruginosa outer membrane vesicles triggered by human mucosal fluid and lysozyme can prime host tissue surfaces for bacterial adhesion

    Directory of Open Access Journals (Sweden)

    Matteo Maria Emiliano Metruccio

    2016-06-01

    Full Text Available Pseudomonas aeruginosa is a leading cause of human morbidity and mortality that often targets epithelial surfaces. Host immunocompromise, or the presence of indwelling medical devices, including contact lenses, can predispose to infection. While medical devices are known to accumulate bacterial biofilms, it is not well understood why resistant epithelial surfaces become susceptible to P. aeruginosa. Many bacteria, including P. aeruginosa, release Outer Membrane Vesicles (OMVs in response to stress that can fuse with host cells to alter their function. Here, we tested the hypothesis that mucosal fluid can trigger OMV release to compromise an epithelial barrier. This was tested using tear fluid and corneal epithelial cells in vitro and in vivo. After 1 h both human tear fluid, and the tear component lysozyme, greatly enhanced OMV release from P. aeruginosa strain PAO1 compared to PBS controls (~100 fold. TEM and SDS-PAGE showed tear fluid and lysozyme-induced OMVs were similar in size and protein composition, but differed from biofilm-harvested OMVs, the latter smaller with fewer proteins. Lysozyme-induced OMVs were cytotoxic to human corneal epithelial cells in vitro and murine corneal epithelium in vivo. OMV exposure in vivo enhanced Ly6G/C expression at the corneal surface, suggesting myeloid cell recruitment, and primed the cornea for bacterial adhesion (~4-fold, P < 0.01. Sonication disrupted OMVs retained cytotoxic activity, but did not promote adhesion, suggesting the latter required OMV-mediated events beyond cell killing. These data suggest that mucosal fluid induced P. aeruginosa OMVs could contribute to loss of epithelial barrier function during medical device-related infections.

  4. Prenatal sex determination by radioimmunoassay of testosterone with and without chromatography of the amniotic fluid

    International Nuclear Information System (INIS)

    Distler, W.; Boniver-Ollmann, U.; Tigges, J.; Terinde, R.; Claussen, U.

    1979-01-01

    Amniotic fluid testosterone measured by radioimmunoassay (RIA) without chromatography (immunoreactive testosterone) seems not to be a definitive test for prenatal sex determination in all cases. In this study testosterone (T) levels measured by RIA with chromatography of the amniotic fluid samples were compared with immunoreactive testosterone (iT) values, to determine the predictive accuracy of the two methods. In 111 amniotic fluid samples between 15 and 19 weeks of gestation iT and T were measured parallelly. There are significant differences between iT- and T-means of both sexes (p [de

  5. Multielemental analysis of 18 essential and toxic elements in amniotic fluid samples by ICP-MS: Full procedure validation and estimation of measurement uncertainty.

    Science.gov (United States)

    Markiewicz, B; Sajnóg, A; Lorenc, W; Hanć, A; Komorowicz, I; Suliburska, J; Kocyłowski, R; Barałkiewicz, D

    2017-11-01

    Amniotic fluid is the substantial factor in the development of an embryo and fetus due to the fact that water and solutes contained in it penetrate the fetal membranes in an hydrostatic and osmotic way as well as being swallowed by the fetus. Elemental composition of amniotic fluid influences the growth and health of the fetus, therefore, an analysis of amniotic fluid is important because the results would indicate abnormal levels of minerals or toxic elements. Inductively coupled plasma mass spectroscopy (ICP-MS) is often used for determination of trace and ultra-trace level elements in a wide range of matrices including biological samples because of its unique analytical capabilities. In the case of trace and ultra-trace level analysis detailed characteristics of analytical procedure as well as properties of the analytical result are particularly important. The purpose of this study was to develop a new analytical procedure for multielemental analysis of 18 elements (Al, As, Ba, Ca, Cd, Co, Cr, Cu, Mg, Mn, Ni, Pb, Sb, Se, Sr, U, V and Zn) in amniotic fluid samples using ICP-MS. Dynamic reaction cell (DRC) with two reaction gases, ammonia and oxygen, was involved in the experiment to eliminate spectral interferences. Detailed validation was conducted using 3 certified reference mterials (CRMs) and real amniotic fluid samples collected from patients. Repeatability for all analyzed analytes was found to range from 0.70% to 8.0% and for intermediate precision results varied from 1.3% to 15%. Trueness expressed as recovery ranged from 80% to 125%. Traceability was assured through the analyses of CRMs. Uncertainty of the results was also evaluated using single-laboratory validation approach. The obtained expanded uncertainty (U) results for CRMs, expressed as a percentage of the concentration of an analyte, were found to be between 8.3% for V and 45% for Cd. Standard uncertainty of the precision was found to have a greater influence on the combined standard uncertainty

  6. Trace elements and antibacterial activity in amniotic fluid.

    Science.gov (United States)

    Honkonen, E; Näntö, V; Hyörä, H; Vuorinen, K; Erkkola, R

    1986-01-01

    Antibacterial activity and trace element concentrations in amniotic fluid (AF) were determined in a population of 39 pregnant women in the second half of gestation. Antibacterial activity in each AF was measured by a spectrophotometric micromethod after 18 h incubation at 37 degrees C using Escherichia coli K 12 as a reference bacterium. Concentrations of zinc, iron, copper, calcium, potassium and bromine were measured by particle-induced X-ray emission method and the zinc concentration was also measured by atomic absorption spectrophotometry. Phosphate concentration was determined by direct albumin adding method. In AFs with good antibacterial activity significantly lower concentrations of potassium and bromine were found when compared to AFs with lower antibacterial activity. Concentrations of zinc, iron, copper, calcium or phosphate did not correlate with antibacterial activity in AF.

  7. Calcium transport in sealed vesicles from red beet (Beta vulgaris L.) storage tissue. II. Characterization of 45Ca2+ uptake into plasma membrane vesicles

    International Nuclear Information System (INIS)

    Giannini, J.L.; Ruiz-Cristin, J.; Briskin, D.P.

    1987-01-01

    Calcium uptake was examined in sealed plasma membrane vesicles isolated from red beet (Beta vulgaris L.) storage tissue using 45 Ca 2+ . Uptake of 45 Ca 2+ by the vesicles was ATP-dependent and radiotracer accumulated by the vesicles could be released by the addition of the calcium ionophore A23187. The uptake was stimulated by gramicidin D but slightly inhibited by carbonylcyanide m-chlorophenylhydrazone. Although the latter result might suggest some degree of indirect coupling of 45 Ca 2+ uptake to ATP utilization via ΔμH + , no evidence for a secondary H + /Ca 2+ antiport in this vesicle system could be found. Following the imposition of an acid-interior pH gradient, proton efflux from the vesicle was not enhanced by the addition of Ca 2+ and an imposed pH gradient could not drive 45 Ca 2+ uptake. Optimal uptake of 45 Ca 2+ occurred broadly between pH 7.0 and 7.5 and the transport was inhibited by orthovanadate, N,N'-dicyclohexylcarbodiimide, and diethylstilbestrol but insensitive to nitrate and azide. The dependence of 45 Ca 2+ uptake on both calcium and Mg:ATP concentration demonstrated saturation kinetics with K/sub m/ values of 6 micromolar and 0.37 millimolar, respectively. While ATP was the preferred substrate for driving 45 Ca 2+ uptake, GTP could drive transport at about 50% of the level observed for ATP. The results of this study demonstrate the presence of a unique primary calcium transport system associated with the plasma membrane which could drive calcium efflux from the plant cell

  8. A study of chromosomal aberrations in amniotic fluid cell cultures.

    Science.gov (United States)

    Wolstenholme, J; Crocker, M; Jonasson, J

    1988-06-01

    This paper represents the analysis of 1916 routine amniotic fluid specimens harvested by an in situ fixation technique in a prospective study with regard to cultural chromosome anomalies. Excluding constitutional abnormalities, 2.9 per cent of 19,432 cells analysed showed some form of chromosome anomaly, terminal deletions (57 per cent) and chromatid/chromosome breaks and gaps (18 per cent) being the most frequent, followed by interchange aberrations (13 per cent) and trisomy (5 per cent). No case was found of more than one colony from the same culture showing the same anomaly without it being present in other cultures from the same fluid. The wholly abnormal colonies had a surplus of trisomies and from the mathematical considerations presented one may infer that these are likely to reflect the presence of abnormal cells in the amniotic fluid. Partly abnormal colonies appeared at a frequency that would correspond to virtual absence of selection against chromosomally abnormal cells when cultured in vitro. The aberrations found were similar to those seen as single cell anomalies, except for chromatid breaks and exchanges. The data suggest a basic preferential induction of trisomy for chromosomes 2, 18, 21, and the Y-chromosome. Structural aberrations showed a marked clustering of breakpoints around the centromeres. The frequency of mutant cells was low (1.4 X 10(-3)) before culture was initiated. At harvest, the frequency of abnormal cells was much higher (3 X 10(-2)) corresponding to 3 X 10(-3) mutations per cell per generation accumulating over approximately ten generations in vitro.

  9. Infection-Induced Thrombin Production: A Potential Novel Mechanism for Preterm Premature Rupture of Membranes (PPROM).

    Science.gov (United States)

    Feng, Liping; Allen, Terrence K; Marinello, William P; Murtha, Amy P

    2018-04-13

    Preterm premature rupture of membranes (PPROM) is a leading contributor to maternal and neonatal morbidity and mortality. Epidemiologic and experimental studies have demonstrated that thrombin causes fetal membrane weakening and subsequently PPROM. Although blood is suspected as the likely source of thrombin in fetal membranes and amniotic fluid of patients with PPROM, this has not been proven. Ureaplasma Parvum (U. parvum) is emerging as a pathogen involved in prematurity, including PPROM, but until now, prothrombin production directly induced by bacteria in fetal membranes has not been described. This study was designed to investigate whether U. parvum exposure can induce prothrombin production in fetal membranes cells. Primary fetal membrane cells (amnion epithelial, chorion trophoblast, and decidua stromal) or full-thickness fetal membrane tissue explants from elective, term, uncomplicated cesarean deliveries were harvested. Cells or tissue explants were infected with live U. parvum (1 x 10 5 , 1 x 10 6 , or 1 x 10 7 colony forming units (cfu)/ml) or lipopolysaccharide (Escherichia coli J5, L-5014, Sigma, 100 ng/ml or 1000 ng/ml) for 24 hours. Tissue explants were fixed for immunohistochemistry staining of thrombin/prothrombin. Fetal membrane cells were fixed for confocal immunofluorescent staining of the biomarkers of fetal membrane cell types and thrombin/prothrombin. Protein and mRNA were harvested from the cells and tissue explants for Western blot or qRT-PCR to quantify thrombin/prothrombin protein or mRNA production, respectively. Data are presented as mean values ± standard errors of mean. Data were analyzed using one-way ANOVA with post hoc Dunnett's test. Prothrombin production and localization was confirmed by Western blot and immunostainings in all primary fetal membrane cells and tissue explants. Immunofluorescence observations revealed a perinuclear localization of prothrombin in amnion epithelial cells. Localization of prothrombin in chorion and

  10. Radioimmunoassay of alpha-foeto protein in the amniotic fluid in normal and pathological pregnancies

    International Nuclear Information System (INIS)

    Degueldre, M.; Golstein, J.; Rodesch, F.; L'Hermite, M.

    1975-01-01

    A radioimmunoassay of AFP was developed and the normal amniotic concentrations of this protein were measured as a function of the gestation age. The specific reagents used included a rabbit anti-AFP serum and a highly purified preparation of AFP labelled with 125 I, which also served as a standard. Amniotic fluid was drawn off by amniocentesis between the 11th and 40th week following the last menstrual period. Samples of amniotic fluid were also obtained in 2 cases of foetal death in utero and 3 cases of anencephaly. The normal amniotic AFP concentration limits were particularly well established between the 14th and 18th weeks, an ideal period to perform an amniocentesis both from a technical viewpoint and with regard to the bulk of information obtainable for the antenatal diagnosis of many congenital diseases. The concentrations observed in pathological cases were distinctly higher than the upper 95% confidence limit of normal values for the gestation age considered [fr

  11. Matrix metalloproteinase-2 is elevated in midtrimester amniotic fluid prior to the development of preeclampsia

    Directory of Open Access Journals (Sweden)

    Daniel-Spiegel Etty

    2009-08-01

    Full Text Available Abstract Objective To evaluate levels of matrix metalloproteinases (MMP and their inhibitors (TIMP in second trimester amniotic fluid of women with hypertensive disorders compared to normotensive women. Study Design Amniotic fluid was obtained from 133 women undergoing genetic second trimester amniocentesis. Zymography was performed for MMP characterization and an MMP-2 ELISA kit was used to determine MMP-2 levels. TIMP-2 expression was evaluated using western blot. Results Mean amniotic fluid MMP-2 and TIMP-2 levels were significantly higher in women who developed a hypertensive disorder compared to normotensive women (P Conclusion Higher amniotic fluid MMP-2 and TIMP-2 levels are found in women who eventually develop preeclampsia.

  12. Prophylactic Cefazolin in Amnioinfusions Administered for Meconium-Stained Amniotic Fluid

    Directory of Open Access Journals (Sweden)

    R. K. Edwards

    1999-01-01

    Full Text Available Objective: To determine if amnioinfusion with an antibiotic solution decreased the rate of clinical chorioamnionitis and puerperal endometritis in patients with meconium-stained amniotic fluid.

  13. Bone morphogenetic protein-7 promotes chondrogenesis in human amniotic epithelial cells.

    Science.gov (United States)

    Zhou, Junjie; Yu, Guangrong; Cao, Chengfu; Pang, Jinhui; Chen, Xianqi

    2011-06-01

    Bone morphogenetic proteins (BMPs) play important roles at multiple stages of chondrogenesis. This study was undertaken to investigate the potential role of bone morphogenetic protein-7 (BMP-7) in the differentiation of chondrocytes using tissue engineering techniques. The impact of BMP-7 on human amniotic epithelial cells (hAECs) was tested. The hAECs were treated either with recombinant human BMP-7 cDNA or with transforming growth factor beta 1 (TGF-β1) as a positive control for three weeks in vitro. Cartilaginous differentiation and proliferation were assayed by quantitative RT-PCR, histology, and in situ hybridization. Our results were such that hAECs treated with either BMP-7 or TGF-β1 expressed cartilage markers (aggrecan, Sox9, CEP-68, and type II and X collagens) within three weeks. Compared with a control vector, BMP-7 induced a decrease in type I collagen expression, while the transcription of the cartilage-specific type II collagen remained stable. In induction experiments, BMP-7 transgenic hAECs exhibited the largest amount of matrix synthesis. In conclusion, these data indicate that BMP-7 plays an important role in inducing the production of cartilage by hAECs in vitro. Cartilage differentiation and matrix maturation can be promoted by BMPs in a cartilage engineering paradigm. These properties make BMPs promising tools in the engineering of cartilaginous joint bio-prostheses and as candidate biological agents or genes for cartilage stabilisation.

  14. Expansion of the neck reconstituted the shoulder-diaphragm in amniote evolution.

    Science.gov (United States)

    Hirasawa, Tatsuya; Fujimoto, Satoko; Kuratani, Shigeru

    2016-01-01

    The neck acquired flexibility through modifications of the head-trunk interface in vertebrate evolution. Although developmental programs for the neck musculoskeletal system have attracted the attention of evolutionary developmental biologists, how the heart, shoulder and surrounding tissues are modified during development has remained unclear. Here we show, through observation of the lateral plate mesoderm at cranial somite levels in chicken-quail chimeras, that the deep part of the lateral body wall is moved concomitant with the caudal transposition of the heart, resulting in the infolding of the expanded cervical lateral body wall into the thorax. Judging from the brachial plexus pattern, an equivalent infolding also appears to take place in mammalian and turtle embryos. In mammals, this infolding process is particularly important because it separates the diaphragm from the shoulder muscle mass. In turtles, the expansion of the cervical lateral body wall affects morphogenesis of the shoulder. Our findings highlight the cellular expansion in developing amniote necks that incidentally brought about the novel adaptive traits. © 2015 The Authors Development, Growth & Differentiation published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Developmental Biologists.

  15. Stem Cells Derived from Amniotic Fluid: A Potential Pluripotent-Like Cell Source for Cellular Therapy?

    Science.gov (United States)

    Ramasamy, Thamil Selvee; Velaithan, Vithya; Yeow, Yelena; Sarkar, Fazlul H

    2018-01-01

    Regenerative medicine aims to provide therapeutic treatment for disease or injury, and cell-based therapy is a newer therapeutic approach different from conventional medicine. Ethical issues that rose by the utilisation of human embryonic stem cells (hESC) and the limited capacity of adult stem cells, however, hinder the application of these stem cells in regenerative medicine. Recently, isolation and characterisation of c-kit positive cells from human amniotic fluid, which possess intermediate characteristics between hESCs and adult stem cells, provided a new approach towards realising their promise for fetal and adult regenerative medicine. Despite the number of studies that have been initiated to characterize their molecular signature, research on developing approaches to maintain and enhance their regenerative potential is urgently needed and must be developed. Thus, this review is focused on understanding their potential uses and factors influencing their pluripotent status in vitro. In short, this cell source could be an ideal cellular resource for pluripotent cells for potential applications in allogeneic cellular replacement therapies, fetal tissue engineering, pharmaceutical screening, and in disease modelling. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  16. The biological activities of (1,3)-(1,6)-{beta}-d-glucan and porous electrospun PLGA membranes containing {beta}-glucan in human dermal fibroblasts and adipose tissue-derived stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Woo, Yeon I; Park, Bong Joo; Kim, Hye-Lee; Lee, Mi Hee; Kim, Jungsung; Park, Jong-Chul [Department of Medical Engineering, Yonsei University College of Medicine, 134 Shinchon-dong, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Yang, Young-Il [Department of Pathology, School of Medicine, Paik Institute for Clinical Research, Inje University, 633-165 Gae-dong, Busan-jin-gu, Busan 614-735 (Korea, Republic of); Kim, Jung Koo [Department of Biomedical Engineering, College of Biomedical Science and Engineering, Inje University, Kimhae 621-749 (Korea, Republic of); Tsubaki, Kazufumi [R and D division, Asahi Denka Co. Ltd, 7-2-35 Higashi-ogu, Arakawa-ku, Tokyo 116-8554 (Japan); Han, Dong-Wook, E-mail: parkjc@yuhs.a [Department of Nanomedical Engineering, College of Nanoscience and Nanotechnology, Pusan National University, geumjeong-gu, Busan 609-735 (Korea, Republic of)

    2010-08-01

    In this study, we investigated the possible roles of (1,3)-(1,6)-{beta}-d-glucan ({beta}-glucan) and porous electrospun poly-lactide-co-glycolide (PLGA) membranes containing {beta}-glucan for skin wound healing, especially their effect on adult human dermal fibroblast (aHDF) and adipose tissue-derived stem cell (ADSC) activation, proliferation, migration, collagen gel contraction and biological safety tests of the prepared membrane. This study demonstrated that {beta}-glucan and porous PLGA membranes containing {beta}-glucan have enhanced the cellular responses, proliferation and migration, of aHDFs and ADSCs and the result of a collagen gel contraction assay also revealed that collagen gels contract strongly after 4 h post-gelation incubation with {beta}-glucan. Furthermore, we confirmed that porous PLGA membranes containing {beta}-glucan are biologically safe for wound healing study. These results indicate that the porous PLGA membranes containing {beta}-glucan interacted favorably with the membrane and the topical administration of {beta}-glucan was useful in promoting wound healing. Therefore, our study suggests that {beta}-glucan and porous PLGA membranes containing {beta}-glucan may be useful as a material for enhancing wound healing.

  17. The cellular prion protein interacts with the tissue non-specific alkaline phosphatase in membrane microdomains of bioaminergic neuronal cells.

    Directory of Open Access Journals (Sweden)

    Myriam Ermonval

    Full Text Available BACKGROUND: The cellular prion protein, PrP(C, is GPI anchored and abundant in lipid rafts. The absolute requirement of PrP(C in neurodegeneration associated to prion diseases is well established. However, the function of this ubiquitous protein is still puzzling. Our previous work using the 1C11 neuronal model, provided evidence that PrP(C acts as a cell surface receptor. Besides a ubiquitous signaling function of PrP(C, we have described a neuronal specificity pointing to a role of PrP(C in neuronal homeostasis. 1C11 cells, upon appropriate induction, engage into neuronal differentiation programs, giving rise either to serotonergic (1C11(5-HT or noradrenergic (1C11(NE derivatives. METHODOLOGY/PRINCIPAL FINDINGS: The neuronal specificity of PrP(C signaling prompted us to search for PrP(C partners in 1C11-derived bioaminergic neuronal cells. We show here by immunoprecipitation an association of PrP(C with an 80 kDa protein identified by mass spectrometry as the tissue non-specific alkaline phosphatase (TNAP. This interaction occurs in lipid rafts and is restricted to 1C11-derived neuronal progenies. Our data indicate that TNAP is implemented during the differentiation programs of 1C11(5-HT and 1C11(NE cells and is active at their cell surface. Noteworthy, TNAP may contribute to the regulation of serotonin or catecholamine synthesis in 1C11(5-HT and 1C11(NE bioaminergic cells by controlling pyridoxal phosphate levels. Finally, TNAP activity is shown to modulate the phosphorylation status of laminin and thereby its interaction with PrP. CONCLUSION/SIGNIFICANCE: The identification of a novel PrP(C partner in lipid rafts of neuronal cells favors the idea of a role of PrP in multiple functions. Because PrP(C and laminin functionally interact to support neuronal differentiation and memory consolidation, our findings introduce TNAP as a functional protagonist in the PrP(C-laminin interplay. The partnership between TNAP and PrP(C in neuronal cells may

  18. Maternal and fetal characteristics associated with meconium-stained amniotic fluid

    DEFF Research Database (Denmark)

    Balchin, Imelda; Whittaker, John C; Lamont, Ronald F

    2011-01-01

    To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF.......To estimate the rates of meconium-stained amniotic fluid (AF) and adverse outcome in relation to gestational age and racial group, and to investigate the predictors of meconium-stained AF....

  19. Ureaplasma parvum serovar 3 multiple banded antigen size variation after chronic intra-amniotic infection/colonization.

    Science.gov (United States)

    Robinson, James W; Dando, Samantha J; Nitsos, Ilias; Newnham, John; Polglase, Graeme R; Kallapur, Suhas G; Pillow, J Jane; Kramer, Boris W; Jobe, Alan H; Payton, Diane; Knox, Christine L

    2013-01-01

    Ureaplasma species are the microorganisms most frequently associated with adverse pregnancy outcomes. The multiple banded antigen (MBA), a surface-exposed lipoprotein, is a key virulence factor of ureaplasmas. The MBA demonstrates size variation, which we have shown previously to be correlated with the severity of chorioamnion inflammation. We aimed to investigate U. parvum serovar 3 pathogenesis in vivo, using a sheep model, by investigating: MBA variation after long term (chronic) and short term (acute) durations of in utero ureaplasma infections, and the severity of chorioamnionitis and inflammation in other fetal tissues. Inocula of 2 × 10(7) colony-forming-units (CFU) of U. parvum serovar 3 (Up) or media controls (C) were injected intra-amniotically into pregnant ewes at one of three time points: day 55 (69d Up, n = 8; C69, n = 4); day 117 (7d Up, n = 8; C7, n = 2); and day 121 (3d Up, n = 8; C3, n = 2) of gestation (term = 145-150d). At day 124, preterm fetuses were delivered surgically. Samples of chorioamnion, fetal lung, and umbilical cord were: (i) snap frozen for subsequent ureaplasma culture, and (ii) fixed, embedded, sectioned and stained by haematoxylin and eosin stain for histological analysis. Selected fetal lung clinical ureaplasma isolates were cloned and filtered to obtain cultures from a single CFU. Passage 1 and clone 2 ureaplasma cultures were tested by western blot to demonstrate MBA variation. In acute durations of ureaplasma infection no MBA variants (3d Up) or very few MBA variants (7d Up) were present when compared to the original inoculum. However, numerous MBA size variants were generated in vivo (alike within contiguous tissues, amniotic fluid and fetal lung, but different variants were present within chorioamnion), during chronic, 69d exposure to ureaplasma infection. For the first time we have shown that the degree of ureaplasma MBA variation in vivo increased with the duration of gestation.

  20. Ureaplasma parvum serovar 3 multiple banded antigen size variation after chronic intra-amniotic infection/colonization.

    Directory of Open Access Journals (Sweden)

    James W Robinson

    Full Text Available Ureaplasma species are the microorganisms most frequently associated with adverse pregnancy outcomes. The multiple banded antigen (MBA, a surface-exposed lipoprotein, is a key virulence factor of ureaplasmas. The MBA demonstrates size variation, which we have shown previously to be correlated with the severity of chorioamnion inflammation. We aimed to investigate U. parvum serovar 3 pathogenesis in vivo, using a sheep model, by investigating: MBA variation after long term (chronic and short term (acute durations of in utero ureaplasma infections, and the severity of chorioamnionitis and inflammation in other fetal tissues. Inocula of 2 × 10(7 colony-forming-units (CFU of U. parvum serovar 3 (Up or media controls (C were injected intra-amniotically into pregnant ewes at one of three time points: day 55 (69d Up, n = 8; C69, n = 4; day 117 (7d Up, n = 8; C7, n = 2; and day 121 (3d Up, n = 8; C3, n = 2 of gestation (term = 145-150d. At day 124, preterm fetuses were delivered surgically. Samples of chorioamnion, fetal lung, and umbilical cord were: (i snap frozen for subsequent ureaplasma culture, and (ii fixed, embedded, sectioned and stained by haematoxylin and eosin stain for histological analysis. Selected fetal lung clinical ureaplasma isolates were cloned and filtered to obtain cultures from a single CFU. Passage 1 and clone 2 ureaplasma cultures were tested by western blot to demonstrate MBA variation. In acute durations of ureaplasma infection no MBA variants (3d Up or very few MBA variants (7d Up were present when compared to the original inoculum. However, numerous MBA size variants were generated in vivo (alike within contiguous tissues, amniotic fluid and fetal lung, but different variants were present within chorioamnion, during chronic, 69d exposure to ureaplasma infection. For the first time we have shown that the degree of ureaplasma MBA variation in vivo increased with the duration of gestation.

  1. Evolution and homology of the astragalus in early amniotes: new fossils, new perspectives.

    Science.gov (United States)

    O'Keefe, F Robin; Sidor, Christian A; Larsson, Hans C E; Maga, Abdoudaye; Ide, Oumarou

    2006-04-01

    The reorganization of the ankle in basal amniotes has long been considered a key innovation allowing the evolution of more terrestrial and cursorial behavior. Understanding how this key innovation arose is a complex problem that largely concerns the homologizing of the amniote astragalus with the various ossifications in the anamniote tarsus. Over the last century, several hypotheses have been advanced homologizing the amniote astragalus with the many ossifications in the ankle of amphibian-grade tetrapods. There is an emerging consensus that the amniote astragalus is a complex structure emerging via the co-ossification of several originally separate elements, but the identities of these elements remain unclear. Here we present new fossil evidence bearing on this contentious question. A poorly ossified, juvenile astragalus of the large captorhinid Moradisaurus grandis shows clear evidence of four ossification centers, rather than of three centers or one center as posited in previous models of astragalus homology. Comparative material of the captorhinid Captorhinikos chozaensis is also interpretable as demonstrating four ossification centers. A new, four-center model for the homology of the amniote astragalus is advanced, and is discussed in the context of the phylogeny of the Captorhinidae in an attempt to identify the developmental transitions responsible for the observed pattern of ossification within this clade. Lastly, the broader implications for amniote phylogeny are discussed, concluding that the neomorphic pattern of astragalus ossification seen in all extant reptiles (including turtles) arose within the clade Diapsida.

  2. An assessment of the accuracy of visual diagnosis of meconium-stained amniotic fluid

    International Nuclear Information System (INIS)

    Sanlialp, C.; Caglar, G.S.; Tapisiz, O.L.; Avsar, A.F.

    2004-01-01

    Objective: The assessment of meconium content in the amniotic fluid depends on visual observation by clinicians at the bedside. The aim of the present study was to compare visual evaluation of meconium-stained amniotic fluid with spectrophotometer evaluation. Study Design: Ten gram of meconium was added to 100 ml of amniotic fluid and mixed. The solution was serially two-fold diluted with amniotic fluid. The serially diluted tubes' absorbance spectrum was measured at 420 nm and thus a standard scale was established. Ninety five samples of meconium- stained amniotic fluid were collected from labouring women and the grade of meconium was deter- mined visually at the bedside. The samples' absorbance spectrum was measured at 420 nm and recorded. Spectrophotometer was considered gold standard and the ranges of optical density in the standard scale was used to test the accuracy of visual categorization of the samples. In the statistical analysis chi-square test was used and significance was p<0.05. Results: The accuracy rate of visual diagnosis of meconium-stained amniotic fluid were found as statistically significant (accuracy rate=54.74%, p<0.001). Visual evaluation was correct in 19.4% of thin, 53.1 % of moderate and 90.6% of thick meconium samples when examined with spectrophotometer. Conclusion: Visually diagnosed thin meconium can be moderate or thick meconium when examined objectively. The visual diagnosis at bedside is not always reliable and should be replaced with an objective method like spectrophotometry. (author)

  3. Crucial Role of Mesangial Cell-derived Connective Tissue Growth Factor in a Mouse Model of Anti-Glomerular Basement Membrane Glomerulonephritis.

    Science.gov (United States)

    Toda, Naohiro; Mori, Kiyoshi; Kasahara, Masato; Ishii, Akira; Koga, Kenichi; Ohno, Shoko; Mori, Keita P; Kato, Yukiko; Osaki, Keisuke; Kuwabara, Takashige; Kojima, Katsutoshi; Taura, Daisuke; Sone, Masakatsu; Matsusaka, Taiji; Nakao, Kazuwa; Mukoyama, Masashi; Yanagita, Motoko; Yokoi, Hideki

    2017-02-13

    Connective tissue growth factor (CTGF) coordinates the signaling of growth factors and promotes fibrosis. Neonatal death of systemic CTGF knockout (KO) mice has hampered analysis of CTGF in adult renal diseases. We established 3 types of CTGF conditional KO (cKO) mice to investigate a role and source of CTGF in anti-glomerular basement membrane (GBM) glomerulonephritis. Tamoxifen-inducible systemic CTGF (Rosa-CTGF) cKO mice exhibited reduced proteinuria with ameliorated crescent formation and mesangial expansion in anti-GBM nephritis after induction. Although CTGF is expressed by podocytes at basal levels, podocyte-specific CTGF (pod-CTGF) cKO mice showed no improvement in renal injury. In contrast, PDGFRα promoter-driven CTGF (Pdgfra-CTGF) cKO mice, which predominantly lack CTGF expression by mesangial cells, exhibited reduced proteinuria with ameliorated histological changes. Glomerular macrophage accumulation, expression of Adgre1 and Ccl2, and ratio of M1/M2 macrophages were all reduced both in Rosa-CTGF cKO and Pdgfra-CTGF cKO mice, but not in pod-CTGF cKO mice. TGF-β1-stimulated Ccl2 upregulation in mesangial cells and macrophage adhesion to activated mesangial cells were decreased by reduction of CTGF. These results reveal a novel mechanism of macrophage migration into glomeruli with nephritis mediated by CTGF derived from mesangial cells, implicating the therapeutic potential of CTGF inhibition in glomerulonephritis.

  4. Multiple facets of sialomucin complex/MUC4, a membrane mucin and erbb2 ligand, in tumors and tissues (Y2K update).

    Science.gov (United States)

    Carraway, K L; Price-Schiavi, S A; Komatsu, M; Idris, N; Perez, A; Li, P; Jepson, S; Zhu, X; Carvajal, M E; Carraway, C A

    2000-01-01

    Sialomucin complex (SMC, MUC4) is a high Mr glycoprotein heterodimer, composed of mucin (ASGP-1) and transmembrane (ASGP-2) subunits. ASGP-2 contains two EGF-like domains and acts as an intramembrane ligand for the receptor tyrosine kinase ErbB2. Transfection studies with SMC DNAs showed that SMC expression could markedly reduce both cell-cell and cell-matrix interactions in vitro and increase the growth of primary tumors and the formation of metastatic foci of human A375 melanoma cells as xenotransplants in nude mice, possibly through the ability to suppress apoptosis. SMC is expressed in most vulnerable epithelia as a protective agent, which is found in both membrane and soluble forms at luminal surfaces and secreted into fluids such as milk and tears. SMC appears to be constitutively expressed by most accessible epithelia, notable exceptions being the mammary gland and uterine luminal epithelium, in which it is tightly regulated during pregnancy. Down-regulation at the luminal uterine surface appears necessary for blastocyst implantation. TGF-b is a potent repressor of SMC expression in the mammary gland and uterus, though by different mechanisms. These combined results suggest that SMC has multiple functions in epithelia and is tightly regulated in those tissues where its special functions are required.

  5. In vitro secretion profiles of interleukin (IL-1beta, IL-6, IL-8, IL-10, and TNF alpha after selective infection with Escherichia coli in human fetal membranes

    Directory of Open Access Journals (Sweden)

    Maida-Claros Rolando

    2007-12-01

    Full Text Available Abstract Background Chorioamniotic membranes infection is a pathologic condition in which an abnormal secretion of proinflammatory cytokines halts fetal immune tolerance. The aim of the present study was to evaluate the functional response of human chorioamniotic membranes, as well as the individual contribution of the amnion and choriodecidua after stimulation with Escherichia coli, a pathogen associated with preterm labor. Methods Explants of chorioamniotic membranes from 10 women (37–40 weeks of gestation were mounted and cultured in a Transwell system, which allowed us to test the amnion and choriodecidua compartments independently. Escherichia coli (1 × 10 6 CFU/mL was added to either the amniotic or the choriodecidual regions or both; after a 24-h incubation, the secretion of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10 in both compartments was measured using a specific ELISA. Data were analyzed by Kruskal-Wallis one-way analysis of variance. Results After stimulation with Escherichia coli, the choriodecidua compartment showed an increase in the secretion of IL-1beta (21-fold, IL-6 (2-fold, IL-8 (6-fold, and IL-10 (37-fold, regardless of which side of the membrane was stimulated; TNFalpha secretion augmented (22-fold also but only when the stimulus was applied simultaneously to both sides. When the amnion was stimulated directly, the level of IL-1beta (13-fold rose significantly; however, the increase in IL-8 secretion was larger (20-fold, regardless of the primary site of infection. TNFalpha secretion in the amnion compartment rose markedly only when Escherichia coli was simultaneously applied to both sides. Conclusion Selective stimulation of fetal membranes with Escherichia coli results in a differential production of IL-1beta, IL-6, TNFalpha, IL-8, and IL-10. These tissues were less responsive when the amnion side was stimulated. This is in agreement with the hypothesis that the choriodecidua may play a primary role during an ascending

  6. Circulating Vascular Basement Membrane Fragments are Associated with the Diameter of the Abdominal Aorta and Their Expression Pattern is Altered in AAA Tissue.

    Science.gov (United States)

    Holsti, Mari; Wanhainen, Anders; Lundin, Christina; Björck, Martin; Tegler, Gustaf; Svensson, Johan; Sund, Malin

    2018-04-12

    Abdominal aortic aneurysm (AAA) is characterised by enhanced proteolytic activity, and extracellular matrix (ECM) remodelling in the vascular wall. Type IV and XVIII collagen/endostatin are structural proteins in vascular basement membrane (VBM), a specialised ECM structure. Here the association between plasma levels of these collagens with the aortic diameter and expansion rate is studied, and their expression in aortic tissue characterised. This was a retrospective population based cohort study. Type IV and XVIII collagen/endostatin were analysed in plasma by ELISA assay in 615 men, divided into three groups based on the aortic diameter: 1) normal aorta ≤ 25 mm, 2) sub-aneurysmal aorta (SAA) 26-29 mm, and 3) AAA ≥ 30 mm. Follow up data were available for 159 men. The association between collagen levels and aortic diameter at baseline, and with the expansion rate at follow up were analysed in ordinal logistic regression and linear regression models, controlling for common confounding factors. Tissue expression of the collagens was analysed in normal aorta (n = 6) and AAA (n = 6) by immunofluorescence. Plasma levels of type XVIII collagen/endostatin (136 ng/mL [SD 29] in individuals with a normal aorta diameter, 154 ng/ml [SD 45] in SAA, and 162 ng/ml [SD 46] in AAA; p = .001) and type IV collagen (105 ng/mL [SD 42] normal aorta, 124 ng/ml [SD 46] SAA, and 127 ng/ml [SD 47] AAA; p = .037) were associated with a larger aortic diameter. A significant association was found between the baseline levels of type XVIII/endostatin and the aortic expansion rate (p = .035), but in the multivariable model, only the initial aortic diameter remained significantly associated with expansion (p = .005). Altered expression patterns of both collagens were observed in AAA tissue. Plasma levels of circulating type IV and XVIII collagen/endostatin increase with AAA diameter. The expression pattern of VBM proteins is altered in the aneurysm wall. Copyright

  7. Transcapillary permeability and subendothelial distribution of endothelial and amniotic fluid insulin-like growth factor binding proteins in the rat heart

    International Nuclear Information System (INIS)

    Bar, R.S.; Clemmons, D.R.; Boes, M.; Busby, W.H.; Booth, B.A.; Dake, B.L.; Sandra, A.

    1990-01-01

    Insulin-like growth factor (IGF) binding proteins (IGFBP) were purified from conditioned media of cultured bovine endothelial cells (ECBP) and from human amniotic fluid (IGFBP-1), and then labeled by radioiodination. 125I-ECBP and 125I-IGFBP-1 were perfused through isolated beating rat hearts for 1 and 5 min, and the hearts fixed and analyzed for 125I-BP content and distribution. One to 4% of the perfused 125I-ECBP and 125I-IGFBP-1 crossed the capillary boundary. The ECBPs predominantly localized as intact 125I-BP in connective tissue elements of the heart with less 125I-BP in cardiac muscle. The ratio of 125I-ECBP in connective tissue: muscle (normalized to percent vol of these compartments) was greater than or equal to 10:1. In contrast, the IGFBP-1 had a greater affinity for cardiac muscle with ratios of 125I-IGFBP-1 in connective tissue:muscle of approximately 1:2. When 125I-IGF-I, in the absence of any BPs, was perfused through the hearts approximately 3-5% left the microcirculation and was found in subendothelial tissues. 125I-IGF-I localized primarily to cardiac muscle with a distribution of connective tissue:cardiac muscle of about 1:3. The findings in the isolated perfused heart were confirmed in intact animals. After 125I-IGFBP-1 was injected into anesthetized rats and allowed to circulate for 5 min, substantial radioactivity was associated with the heart. As in the isolated heart, the IGFBP-1 preferentially localized to cardiac muscle with a connective tissue:cardiac muscle ratio of 1:3. We conclude that IGFBPs produced by endothelial cells and the IGFBP-1 contained in amniotic fluid can cross the capillary boundaries of the rat heart, and that the ECBPs preferentially localize in connective tissue elements of the myocardium, whereas IGFBP-1 predominantly localizes in cardiac muscle

  8. A comparative clinico-radiographic study of guided tissue regeneration with bioresorbable membrane and a composite synthetic bone graft for the treatment of periodontal osseous defects

    Directory of Open Access Journals (Sweden)

    Sumedha Srivastava

    2015-01-01

    Full Text Available Aim: The aim was to evaluate the bonefill in periodontal osseous defects with the help of guided tissue regeneration, bioresorbable membrane (PerioCol + bone graft (Grabio Glascera in combination and with bonegraft (Grabio Glascera alone. Materials and Methods: The study involved total 30 sites in systemically healthy 19 patients. The parameters for evaluation includes plaque index sulcus bleeding index with one or more periodontal osseous defects having (i probing depth (PD of ≥ 5 mm (ii clinical attachment loss (CAL of ≥ 5 mm and (iii ≥3 mm of radiographic periodontal osseous defect (iv bonefill (v crestal bone loss (vi defect resolution. The study involved the three wall and two wall defects which should be either located interproximally or involving the furcation area. The statistical analysis was done using Statistical Package for Social Sciences, the Wilcoxon signed rank statistic W + for Mann-Whitney U-test. Results: The net gain in PD and CAL after 6 months for Group I ([PerioCol] + [Grabio Glascera] and Group II (Grabio Glascera was 3.94 ± 1.81 mm, 3.57 ± 2.21 mm and 3.94 ± 1.81, 3.57 ± 2.21 mm, respectively. The results of the study for Group I and Group II with regards to mean net bonefill, was 3.25 ± 2.32 (58% mm and 5.14 ± 3.84 (40.26 ± 19.14% mm, crestal bone loss − 0.25 ± 0.68 mm and − 0.79 ± 1.19 mm. Defect resolution 3.50 ± 2.34 mm and 5.93 ± 4.01 mm, respectively. Conclusion: On comparing both the groups together after 6 months of therapy, the results were equally effective for combination of graft and membrane versus bone graft alone since no statistical significant difference was seen between above parameters for both the groups. Thus, both the treatment modalities are comparable and equally effective.

  9. High Aminopeptidase N/CD13 Levels Characterize Human Amniotic Mesenchymal Stem Cells and Drive Their Increased Adipogenic Potential in Obese Women

    Science.gov (United States)

    Iaffaldano, Laura; Nardelli, Carmela; Raia, Maddalena; Mariotti, Elisabetta; Ferrigno, Maddalena; Quaglia, Filomena; Labruna, Giuseppe; Capobianco, Valentina; Capone, Angela; Maruotti, Giuseppe Maria; Pastore, Lucio; Di Noto, Rosa; Martinelli, Pasquale; Del Vecchio, Luigi

    2013-01-01

    Maternal obesity is associated to increased fetal risk of obesity and other metabolic diseases. Human amniotic mesenchymal stem cells (hA-MSCs) have not been characterized in obese women. The aim of this study was to isolate and compare hA-MSC immunophenotypes from obese (Ob-) and normal weight control (Co-) women, to identify alterations possibly predisposing the fetus to obesity. We enrolled 16 Ob- and 7 Co-women at delivery (mean/SEM prepregnancy body mass index: 40.3/1.8 and 22.4/1.0 kg/m2, respectively), and 32 not pregnant women. hA-MSCs were phenotyped by flow cytometry; several maternal and newborn clinical and biochemical parameters were also measured. The expression of membrane antigen CD13 was higher on Ob-hA-MSCs than on Co-hA-MSCs (P=0.005). Also, serum levels of CD13 at delivery were higher in Ob- versus Co-pregnant women and correlated with CD13 antigen expression on Ob-hA-MSCs (r2=0.84, P<0.0001). Adipogenesis induction experiments revealed that Ob-hA-MSCs had a higher adipogenic potential than Co-hA-MSCs as witnessed by higher peroxisome proliferator-activated receptor gamma and aP2 mRNA levels (P=0.05 and P=0.05, respectively), at postinduction day 14 associated with increased CD13 mRNA levels from baseline to day 4 postinduction (P<0.05). Adipogenesis was similar in the two sets of hA-MSCs after CD13 silencing, whereas it was increased in Co-hA-MSCs after CD13 overexpression. CD13 expression was high also in Ob-h-MSCs from umbilical cords or visceral adipose tissue of not pregnant women. In conclusion, antigen CD13, by influencing the adipogenic potential of hA-MSCs, could be an in utero risk factor for obesity. Our data strengthen the hypothesis that high levels of serum and MSC CD13 are obesity markers. PMID:23488598

  10. Towards an understanding of the evolution of the chorioallantoic placenta: steroid biosynthesis and steroid hormone signaling in the chorioallantoic membrane of an oviparous reptile.

    Science.gov (United States)

    Cruze, Lori; Kohno, Satomi; McCoy, Michael W; Guillette, Louis J

    2012-09-01

    Amniotes, mammals, reptiles, and birds form common extraembryonic membranes during development to perform essential functions, such as protection, nutrient transfer, gas exchange, and waste removal. Together with the maternal uterus, extraembryonic membranes of viviparous (live-bearing) amniotes develop as an endocrine placenta that synthesizes and responds to steroid hormones critical for development. The ability of these membranes to synthesize and respond to steroid hormone signaling has traditionally been considered an innovation of placental amniotes. However, our laboratory recently demonstrated that this ability extends to the chorioallantoic membrane (CAM) of an oviparous (egg-laying) amniote, the domestic chicken, and we hypothesized that steroidogenic extraembryonic membranes could be an evolutionarily conserved characteristic of all amniotes because of similarities in basic structure, function, and shared evolutionary ancestry. In this study, we examined steroid hormone synthesis and signaling in the CAM of another oviparous amniote, the American alligator (Alligator mississippiensis). We quantified mRNA expression of a steroidogenic factor involved in the regulation of steroidogenesis (NR5A1), the key steroidogenic enzymes involved in the synthesis of progestins (HSD3B1), androgens (CYP17A1), and estrogens (CYP19A1), and the receptors involved in the signaling of progestins (PR), androgens (AR), estrogens (ESR1 and ESR2), and glucocorticoids (GR). Furthermore, we performed protein immunolocalization for PR and ESR1. Collectively, our findings indicate that the alligator CAM has the capability to regulate, synthesize, and respond to steroid hormone signaling, thus, supporting our hypothesis that the extraembryonic membranes of Amniota share a unifying characteristic, that is, the ability to synthesize and respond to steroid hormones.

  11. Intrapartum transcervical amnioinfusion for meconium-stained amniotic fluid.

    Science.gov (United States)

    Das, A K; Jana, N; Dasgupta, S; Samanta, B

    2007-06-01

    To assess the rates of cesarean deliveries and perinatal outcome following intrapartum transcervical amnioinfusion in women with meconium-stained amniotic fluid (MSAF) in a setting with no electronic fetal monitoring or specialized neonatal care. In this prospective comparative study with 150 women who were in labor and had MSAF, 50 of the women received a transcervical amnioinfusion and the remaining 100 women received standard care. The inclusion criteria were a pregnancy of at least 37 weeks' duration, a single live fetus in cephalic presentation, no major medical or obstetric complications, and no known fetal malformation. The amnioinfusion was performed with 1000 mL of normal saline solution through a red rubber catheter. Amnioinfusion was associated with a significant decrease in the incidence of low Apgar score (amnioinfusion than in the control group (24% vs. 48%; RR, 0.5; 95% CI, 0.29-0.85). There were no major complications related to amnioinfusion. Intrapartum amnioinfusion for MSAF is a simple, safe, effective, and inexpensive procedure feasible in settings where intrapartum monitoring is limited. It is associated with improved perinatal outcome and could lower cesarean delivery rates in low-resource countries.

  12. The role of the notochord in amniote vertebral column segmentation.

    Science.gov (United States)

    Ward, Lizzy; Pang, Angel S W; Evans, Susan E; Stern, Claudio D

    2018-07-01

    The vertebral column is segmented, comprising an alternating series of vertebrae and intervertebral discs along the head-tail axis. The vertebrae and outer portion (annulus fibrosus) of the disc are derived from the sclerotome part of the somites, whereas the inner nucleus pulposus of the disc is derived from the notochord. Here we investigate the role of the notochord in vertebral patterning through a series of microsurgical experiments in chick embryos. Ablation of the notochord causes loss of segmentation of vertebral bodies and discs. However, the notochord cannot segment in the absence of the surrounding sclerotome. To test whether the notochord dictates sclerotome segmentation, we grafted an ectopic notochord. We find that the intrinsic segmentation of the sclerotome is dominant over any segmental information the notochord may possess, and no evidence that the chick notochord is intrinsically segmented. We propose that the segmental pattern of vertebral bodies and discs in chick is dictated by the sclerotome, which first signals to the notochord to ensure that the nucleus pulposus develops in register with the somite-derived annulus fibrosus. Later, the notochord is required for maintenance of sclerotome segmentation as the mature vertebral bodies and intervertebral discs form. These results highlight differences in vertebral development between amniotes and teleosts including zebrafish, where the notochord dictates the segmental pattern. The relative importance of the sclerotome and notochord in vertebral patterning has changed significantly during evolution. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  13. Atypical Amniotic Fluid Embolism Managed with a Novel Therapeutic Regimen

    Directory of Open Access Journals (Sweden)

    Shadi Rezai

    2017-01-01

    Full Text Available Amniotic fluid embolism (AFE is the second leading cause of maternal mortality in the USA with an incidence of 1 : 15,200 births. The case fatality rate and perinatal mortality associated with AFE are 13–30% and 9–44%, respectively. This rare but devastating complication can be difficult to diagnose as many of the early signs and symptoms are nonspecific. Compounding this diagnostic challenge is a lack of effective treatment regimens which to date are mostly supportive. We present the case of a 26-year-old woman who suffered from suspected AFE and was successfully treated with the novel regimen of Atropine, Ondansetron, and Ketorolac (A-OK. The authors acknowledge that this case does not meet the new criteria proposed, by Clark in 2016, but feel that it is important to share this case report, due to dramatic patient response to the provided supportive therapy presented in this case report. We hope this case report will prompt further research into this novel approach to treating AFE with Atropine, Ondansetron, and Ketorolac.

  14. Biochemical composition of amniotic fluid in normal puppies at term of pregnancy: preliminary data

    Directory of Open Access Journals (Sweden)

    Barbara Bolis

    2017-05-01

    Full Text Available The full knowledge of the normal fetal fluids composition could be useful in the dog for the better management of newborns. The aim of the present study was to define the biochemical composition of amniotic fluid of puppies born by elective Caesarean section (CS at term of pregnancy. The study enrolled 24 purebred bitches, classified into small size (20kg. All the bitches were healthy and clinically monitored from mating until parturition. For all the bitches an elective CS at term of pregnancy was performed [1]. For each puppy, the amniotic fluid was collected, immediately centrifuged and frozen at – 20° C until analysis for ALB, AMY, TB, CHOL, CK, ALP, GGT, AST, ALT, LDH, Mg, Ca, K, Na, Trig, BUN, Glc, TP, CREA, LIP, Cl, and GLOB. Data were analyzed by ANCOVA to verify the possible effects of parity, breed body size and newborn gender on amniotic biochemical composition. A total of 69 amniotic fluid samples were collected. The amniotic mean±SD and min-max values for each parameter were defined. LDH (p<0.01 and CK activity (p<0.05, as well as Glc concentrations (p<0.0001 were negatively influenced by the parity. AMY activity was significantly (p<0.05 higher in large sized (44.2±20.87 U/L respect to small sized dogs (30.3±19.89 U/L, while lower (p<0.05 CHOL amniotic concentrations were found in small sized (3.0±2.71 mg/dl as compared to large sized (3.9±2.93 mg/dl dogs. Gender of the newborn did not influence the amniotic biochemical composition. The preliminary results of this study showed some similarities as well as some differences concerning the biochemical composition of the amniotic fluid in dogs at term of pregnancy if compared to data reported for the cat [2]. Furthermore, the results suggested that, in dogs, some amniotic parameters could be influenced by breed body size and by parity.

  15. Synergic activation of toll-like receptor (TLR) 2/6 and 9 in response to Ureaplasma parvum & urealyticum in human amniotic epithelial cells.

    Science.gov (United States)

    Triantafilou, Martha; De Glanville, Benjamin; Aboklaish, Ali F; Spiller, O Brad; Kotecha, Sailesh; Triantafilou, Kathy

    2013-01-01

    Ureaplasma species are the most frequently isolated microorganisms inside the amniotic cavity and have been associated with spontaneous abortion, chorioamnionitis, premature rupture of the membranes (PROM), preterm labour (PL) pneumonia in neonates and bronchopulmonary dysplasia in neonates. The mechanisms by which Ureaplasmas cause such diseases remain unclear, but it is believed that inappropriate induction of inflammatory responses is involved, triggered by the innate immune system. As part of its mechanism of activation, the innate immune system employs germ-lined encoded receptors, called pattern recognition receptors (PRRs) in order to "sense" pathogens. One such family of PRRs are the Toll like receptor family (TLR). In the current study we aimed to elucidate the role of TLRs in Ureaplasma-induced inflammation in human amniotic epithelial cells. Using silencing, as well as human embryonic kidney (HEK) transfected cell lines, we demonstrate that TLR2, TLR6 and TLR9 are involved in the inflammatory responses against Ureaplasma parvum and urealyticum serovars. Ureaplasma lipoproteins, such as Multiple Banded antigen (MBA), trigger responses via TLR2/TLR6, whereas the whole bacterium is required for TLR9 activation. No major differences were observed between the different serovars. Cell activation by Ureaplasma parvum and urealyticum seem to require lipid raft function and formation of heterotypic receptor complexes comprising of TLR2 and TLR6 on the cell surface and TLR9 intracellularly.

  16. Synergic activation of toll-like receptor (TLR 2/6 and 9 in response to Ureaplasma parvum & urealyticum in human amniotic epithelial cells.

    Directory of Open Access Journals (Sweden)

    Martha Triantafilou

    Full Text Available Ureaplasma species are the most frequently isolated microorganisms inside the amniotic cavity and have been associated with spontaneous abortion, chorioamnionitis, premature rupture of the membranes (PROM, preterm labour (PL pneumonia in neonates and bronchopulmonary dysplasia in neonates. The mechanisms by which Ureaplasmas cause such diseases remain unclear, but it is believed that inappropriate induction of inflammatory responses is involved, triggered by the innate immune system. As part of its mechanism of activation, the innate immune system employs germ-lined encoded receptors, called pattern recognition receptors (PRRs in order to "sense" pathogens. One such family of PRRs are the Toll like receptor family (TLR. In the current study we aimed to elucidate the role of TLRs in Ureaplasma-induced inflammation in human amniotic epithelial cells. Using silencing, as well as human embryonic kidney (HEK transfected cell lines, we demonstrate that TLR2, TLR6 and TLR9 are involved in the inflammatory responses against Ureaplasma parvum and urealyticum serovars. Ureaplasma lipoproteins, such as Multiple Banded antigen (MBA, trigger responses via TLR2/TLR6, whereas the whole bacterium is required for TLR9 activation. No major differences were observed between the different serovars. Cell activation by Ureaplasma parvum and urealyticum seem to require lipid raft function and formation of heterotypic receptor complexes comprising of TLR2 and TLR6 on the cell surface and TLR9 intracellularly.

  17. Sealing of Corneal Lacerations Using Photo-Activated Rose Bengal Dye and Amniotic Membrane

    Science.gov (United States)

    2017-01-10

    there were no 213 statistical differences in any of the categories except epithelial hyperplasia . On day 3 there was 214 more epithelial hyperplasia ...histopathological analysis of the three surgical arms was the 273 epithelial hyperplasia noted on day 3. The authors hypothesize that this reflects the... epithelial ingrowth, infection, and potential loss of the eye. Complex lacerations can be difficult 23 to treat, and repair with sutures alone is often

  18. Sealing of Corneal Lacerations Using Photo-Activated Rose Bengal Dye and Amniotic Membrane

    Science.gov (United States)

    2017-05-05

    presentation, state the type (e.g., case report, QA/QI study, program evaluation study, informational report/briefing, etc.) in the "Protocol...Number of Control 19 15 15 Eyes 3mm linear central cornea cornea corneoscleral cornea Experimental Repair 1-Epi Debridement 1-Epi Debridement 1-Epi... Debridement 1-Epi Debridement Method 2-Wound 2-Wound 2-Wound 2-Wound 3-Central 10-0 nylon 3-Apex10-0 nylon 3-Limbal 10-0 nylon 3-RBAM 4-RBAM 4

  19. Use of conjunctival autograft and amniotic membrane in primary pterygium surgery

    OpenAIRE

    Fernández García, Keyly; Gómez Castillo, Zulema; Castillo Pérez, Alexeide; Pérez Parra, Zaadia; Jareño Ochoa, Madelyn; Perea Ruiz, Carlos Alberto

    2012-01-01

    Objetivo: comparar la efectividad del autoinjerto conjuntival y el injerto de membrana amniótica en la cirugía del pterigión primario en el Instituto Cubano de Oftalmología "Ramón Pando Ferrer" de septiembre de 2009 a septiembre de 2010. Métodos: estudio prospectivo, comparativo y aleatorio. El universo estuvo constituido por 80 pacientes (80 ojos) con diagnóstico de pterigión primario que se les realizó escisión quirúrgica de la hiperplasia fibrovascular nasal primaria. Se dividieron en 2 gr...

  20. Sealing of Corneal Lacerations Using Photo Activated Rose Bengal Dye and Amniotic Membrane

    Science.gov (United States)

    2017-04-22

    detected nor was there any appearance o f fluorescein leakage from 234 the retinal vessels (Figure 4 B). 235 Irreversible damage to the photoreceptors...unaffected and no evidence of 35 breakdown of the blood retina barrier was detected . Retina from grccnlight laser treated eyes 36 showed normal RPE...formation of a 55 water -tight seal is critical to preventing infection and preventing surface epithelium from gaining 56 entry into the eye. This stabi

  1. Transabdominal amnioinfusion for improving fetal outcomes after oligohydramnios secondary to preterm prelabour rupture of membranes before 26 weeks

    NARCIS (Netherlands)

    van Teeffelen, Stijn; Pajkrt, Eva; Willekes, Christine; van Kuijk, Sander M. J.; Mol, Ben Willem J.

    2013-01-01

    Preterm prelabour rupture of membranes (PPROM) before 26 weeks can delay lung development and can cause pulmonary hypoplasia, as a result of oligohydramnios. Restoring the amniotic fluid volume by transabdominal amnioinfusion might prevent abnormal lung development and might have a protective effect

  2. Improvement of Heart Failure by Human Amniotic Mesenchymal Stromal Cell Transplantation in Rats.

    Science.gov (United States)

    Razavi Tousi, Seyed Mohammad Taghi; Faghihi, Mahdieh; Nobakht, Maliheh; Molazem, Mohammad; Kalantari, Elham; Darbandi Azar, Amir; Aboutaleb, Nahid

    2016-07-06

    Background: Recently, stem cells have been considered for the treatment of heart diseases, but no marked improvement has been recorded. This is the first study to examine the functional and histological effects of the transplantation of human amniotic mesenchymal stromal cells (hAMSCs) in rats with heart failure (HF). Methods: This study was conducted in the years 2014 and 2015. 35 male Wistar rats were randomly assigned into 5 equal experimental groups (7 rats each) as 1- Control 2- Heart Failure (HF) 3- Sham 4- Culture media 5- Stem Cell Transplantation (SCT). Heart failure was induced using 170 mg/kg/d of isoproterenol subcutaneously injection in 4 consecutive days. The failure confirmed by the rat cardiac echocardiography on day 28. In SCT group, 3×10 6 cells in 150 µl of culture media were transplanted to the myocardium. At the end, echocardiographic and hemodynamic parameters together with histological evaluation were done. Results: Echocardiography results showed that cardiac ejection fraction in HF group increased from 58/73 ± 9% to 81/25 ± 6/05% in SCT group (p value < 0.001). Fraction shortening in HF group was increased from 27/53 ± 8/58% into 45/55 ± 6/91% in SCT group (p value < 0.001). Furthermore, hAMSCs therapy significantly improved mean diastolic blood pressure, mean arterial pressure, left ventricular systolic pressure, rate pressure product, and left ventricular end-diastolic pressure compared to those in the HF group, with the values reaching the normal levels in the control group. A marked reduction in fibrosis tissue was also found in the SCT group (p value < 0.001) compared with the animals in the HF group. Conclusion: The transplantation of hAMSCs in rats with heart failure not only decreased the level of fibrosis but also conferred significant improvement in heart performance in terms of echocardiographic and hemodynamic parameters.

  3. The Effect of Cryopreserved Human Placental Tissues on Biofilm Formation of Wound-Associated Pathogens.

    Science.gov (United States)

    Mao, Yong; Singh-Varma, Anya; Hoffman, Tyler; Dhall, Sandeep; Danilkovitch, Alla; Kohn, Joachim

    2018-01-08

    Biofilm, a community of bacteria, is tolerant to antimicrobial agents and ubiquitous in chronic wounds. In a chronic DFU (Diabetic Foot Ulcers) clinical trial, the use of a human cryopreserved viable amniotic membrane (CVAM) resulted in a high rate of wound closure and reduction of wound-related infections. Our previous study demonstrated that CVAM possesses intrinsic antimicrobial activity against a spectrum of wound-associated bacteria under planktonic culture conditions. In this study, we evaluated the effect of CVAM and cryopreserved viable umbilical tissue (CVUT) on biofilm formation of S. aureus and P. aeruginosa , the two most prominent pathogens associated with chronic wounds. Firstly, we showed that, like CVAM, CVUT released antibacterial activity against multiple bacterial pathogens and the devitalization of CVUT reduced its antibacterial activity. The biofilm formation was then measured using a high throughput method and an ex vivo porcine dermal tissue model. We demonstrate that the formation of biofilm was significantly reduced in the presence of CVAM- or CVUT-derived conditioned media compared to control assay medium. The formation of P. aeruginosa biofilm on CVAM-conditioned medium saturated porcine dermal tissues was reduced 97% compared with the biofilm formation on the control medium saturated dermal tissues. The formation of S. auerus biofilm on CVUT-conditioned medium saturated dermal tissues was reduced 72% compared with the biofilm formation on the control tissues. This study is the first to show that human cryopreserved viable placental tissues release factors that inhibit biofilm formation. Our results provide an explanation for the in vivo observation of their ability to support wound healing.

  4. The Effect of Cryopreserved Human Placental Tissues on Biofilm Formation of Wound-Associated Pathogens

    Directory of Open Access Journals (Sweden)

    Yong Mao

    2018-01-01

    Full Text Available Biofilm, a community of bacteria, is tolerant to antimicrobial agents and ubiquitous in chronic wounds. In a chronic DFU (Diabetic Foot Ulcers clinical trial, the use of a human cryopreserved viable amniotic membrane (CVAM resulted in a high rate of wound closure and reduction of wound-related infections. Our previous study demonstrated that CVAM possesses intrinsic antimicrobial activity against a spectrum of wound-associated bacteria under planktonic culture conditions. In this study, we evaluated the effect of CVAM and cryopreserved viable umbilical tissue (CVUT on biofilm formation of S. aureus and P. aeruginosa, the two most prominent pathogens associated with chronic wounds. Firstly, we showed that, like CVAM, CVUT released antibacterial activity against multiple bacterial pathogens and the devitalization of CVUT reduced its antibacterial activity. The biofilm formation was then measured using a high throughput method and an ex vivo porcine dermal tissue model. We demonstrate that the formation of biofilm was significantly reduced in the presence of CVAM- or CVUT-derived conditioned media compared to control assay medium. The formation of P. aeruginosa biofilm on CVAM-conditioned medium saturated porcine dermal tissues was reduced 97% compared with the biofilm formation on the control medium saturated dermal tissues. The formation of S. auerus biofilm on CVUT-conditioned medium saturated dermal tissues was reduced 72% compared with the biofilm formation on the control tissues. This study is the first to show that human cryopreserved viable placental tissues release factors that inhibit biofilm formation. Our results provide an explanation for the in vivo observation of their ability to support wound healing.

  5. Amniotic fluid gamma-glutamyl transpeptidase activity during the second trimester.

    Science.gov (United States)

    Legge, M; Potter, H C

    1986-03-12

    Gamma glutamyl transpeptidase (GGTP) activity was determined in second trimester amniotic fluid taken from normal fetuses and those with fetal abnormalities. GGTP activity decreased with advancing gestation. Increasing meconium contamination correlated with an increase in GGTP activity as did increasing fetal blood contamination. Maternal blood did not affect GGTP activity. Anencephaly did not significantly alter the GGTP activity, however, fetuses with spina bifida had significantly lower activity. Klinefelters and Turners syndromes both had GGTP activity close to the 50th percentile, and two trisomy 21 fetuses had GGTP activity below the 40th percentile. Two trisomy 18 fetuses and two translocation Downs syndromes (46 XY, t (14;21) had GGTP activities considerably lower than the 20th percentile as did a fetus with gastroschisis. Second trimester amniotic fluid GGTP activity may provide an easy preliminary test to screen amniotic fluids for the possibility of certain fetal chromosome abnormalities.

  6. Amniotic Constriction Bands: Secondary Deformities and Their Treatments.

    Science.gov (United States)

    Drury, Benjamin T; Rayan, Ghazi M

    2018-01-01

    The purpose of this study was to report the surgical treatment experience of patients with amniotic constriction bands (ACB) over a 35-year interval and detail consequential limb deformities with emphasis on hands and upper extremities, along with the nature and frequency of their surgical treatment methods. Fifty-one patients were identified; 26 were males and 25 females. The total number of deformities was listed. The total number of operations, individual procedures, and operations plus procedures that were done for each patient and their frequency were recorded. The total number of operations was 117, and total number of procedures was 341. More procedures were performed on the upper extremity (85%) than the lower extremity (15%). Including the primary deformity ACB, 16 different hand deformities secondary to ACB were encountered. Sixteen different surgical methods for the upper extremity were utilized; a primary procedure for ACB and secondary reconstructions for all secondary deformities. Average age at the time of the first procedure was 9.3 months. The most common procedures performed, in order of frequency, were excision of ACB plus Z-plasty, release of partial syndactyly, release of fenestrated syndactyly, full-thickness skin grafts, resection of digital bony overgrowth from amputation stumps, and deepening of first and other digital web spaces. Many hand and upper extremity deformities secondary to ACB are encountered. Children with ACB may require more than one operation including multiple procedures. Numerous surgical methods of reconstruction for these children's secondary deformities are necessary in addition to the customary primary procedure of excision of ACB and Z-plasty.

  7. Membrane dynamics

    DEFF Research Database (Denmark)

    Bendix, Pól Martin

    2015-01-01

    Current topics include membrane-protein interactions with regard to membrane deformation or curvature sensing by BAR domains. Also, we study the dynamics of membrane tubes of both cells and simple model membrane tubes. Finally, we study membrane phase behavior which has important implications...... for the lateral organization of membranes as wells as for physical properties like bending, permeability and elasticity...

  8. Lim Mineralization Protein 3 Induces the Osteogenic Differentiation of Human Amniotic Fluid Stromal Cells through Kruppel-Like Factor-4 Downregulation and Further Bone-Specific Gene Expression

    Directory of Open Access Journals (Sweden)

    Marta Barba

    2012-01-01

    Full Text Available Multipotent mesenchymal stem cells with extensive self-renewal properties can be easily isolated and rapidly expanded in culture from small volumes of amniotic fluid. These cells, namely, amniotic fluid-stromal cells (AFSCs, can be regarded as an attractive source for tissue engineering purposes, being phenotypically and genetically stable, plus overcoming all the safety and ethical issues related to the use of embryonic/fetal cells. LMP3 is a novel osteoinductive molecule acting upstream to the main osteogenic pathways. This study is aimed at delineating the basic molecular events underlying LMP3-induced osteogenesis, using AFSCs as a cellular model to focus on the molecular features underlying the multipotency/differentiation switch. For this purpose, AFSCs were isolated and characterized in vitro and transfected with a defective adenoviral vector expressing the human LMP3. LMP3 induced the successful osteogenic differentiation of AFSC by inducing the expression of osteogenic markers and osteospecific transcription factors. Moreover, LMP3 induced an early repression of the kruppel-like factor-4, implicated in MSC stemness maintenance. KLF4 repression was released upon LMP3 silencing, indicating that this event could be reasonably considered among the basic molecular events that govern the proliferation/differentiation switch during LMP3-induced osteogenic differentiation of AFSC.

  9. Cell fusion phenomena detected after in utero transplantation of Ds-red-harboring porcine amniotic fluid stem cells into EGFP transgenic mice.

    Science.gov (United States)

    Peng, Shao-Yu; Chen, Yu-Hsu; Chou, Chih-Jen; Wang, Yao-Horng; Lee, Hung-Maan; Cheng, Winston Teng-Kui; Shaw, S W Steven; Wu, Shinn-Chih

    2014-05-01

    Amniotic fluid stem cells (AFSCs) are derived from the amniotic fluid of the developing fetus and can give rise to diverse differentiated cells of ectoderm, mesoderm, and endoderm lineages. Intrauterine transplantation is an approach used to cure inherited genetic fetal defects during the gestation period of pregnant dams. Certain disease such as osteogenesis imperfecta was successfully treated in affected fetal mice using this method. However, the donor cell destiny remains uncertain. The purpose of this study was to investigate the biodistribution and cell fate of Ds-red-harboring porcine AFSCs (Ds-red pAFSCs) after intrauterine transplantation into enhanced green fluorescent protein-harboring fetuses of pregnant mice. Pregnant mice (12.5 days) underwent open laparotomy with intrauterine pAFSC transplantation (5 × 10(4) cells per pup) into fetal peritoneal cavity. Three weeks after birth, the mice were sacrificed. Several samples from different organs were obtained for histological examination and flow cytometric analysis. Ds-red pAFSCs migrated most frequently into the intestines. Furthermore, enhanced green fluorescent protein and red fluorescent protein signals were co-expressed in the intestine and liver cells via immunohistochemistry studies. In utero xenotransplantation of pAFSCs fused with recipient intestinal cells instead of differentiating or maintaining the undifferentiated status in the tissue. © 2014 John Wiley & Sons, Ltd.

  10. Utilization of human amniotic mesenchymal cells as feeder layers to sustain propagation of human embryonic stem cells in the undifferentiated state.

    Science.gov (United States)

    Zhang, Kehua; Cai, Zhe; Li, Yang; Shu, Jun; Pan, Lin; Wan, Fang; Li, Hong; Huang, Xiaojie; He, Chun; Liu, Yanqiu; Cui, Xiaohui; Xu, Yang; Gao, Yan; Wu, Liqun; Cao, Shanxia; Li, Lingsong

    2011-08-01

    Human embryonic stem (ES) cells are usually maintained in the undifferentiated state by culturing on feeder cells layers of mouse embryonic fibroblasts (MEFs). However, MEFs are not suitable to support human ES cells used for clinical purpose because of risk of zoonosis from animal cells. Therefore, human tissue-based feeder layers need to be developed for human ES cells for clinical purpose. Hereof we report that human amniotic mesenchymal cells (hAMCs) could act as feeder cells for human ES cells, because they are easily obtained and relatively exempt from ethical problem. Like MEFs, hAMCs could act as feeder cells for human ES cells to grow well on. The self-renewal rate of human ES cells cultured on hAMCs feeders was higher than that on MEFs and human amniotic epithelial cells determined by measurement of colonial diameters and growth curve as well as cell cycle analysis. Both immunofluorescence staining and immunoblotting showed that human ES cells cultured on hAMCs expressed stem cell markers such as Oct-3/4, Sox2, and NANOG. Verified by embryoid body formation in vitro and teratoma formation in vivo, we found out that after 20 passages of culture, human ES cells grown on hAMCs feeders could still retain the potency of differentiating into three germ layers. Taken together, our data suggested hAMCs may be safe feeder cells to sustain the propagation of human ES cells in undifferentiated state for future therapeutic use.

  11. Long-term amnioinfusion through a subcutaneously implanted amniotic fluid replacement port system for treatment of PPROM in humans.

    Science.gov (United States)

    Tchirikov, Michael; Steetskamp, Joscha; Hohmann, Manfred; Koelbl, Heinz

    2010-09-01

    To introduce a novel method for the treatment of PPROM (preterm premature rupture of membranes) using continuous amnioinfusion via a subcutaneously implanted port system. After development and testing since 2001 in a fetal sheep model, the port system has been successfully implanted in two humans with PPROM. In the first case, the subcutaneous port system was implanted during the 23rd week of gestation in a 39-year-old 5th-gravida with PPROM since the 18th week of gestation; in the second case, the port system was implanted during the 24th week of gestation in a 27-year-old 3rd gravida with PPROM since the 21st week of gestation. After port implantation, 100ml/h saline solution was infused intermittently into the amniotic cavity. The whole course of treatment was supported by tocolysis. In the cases presented, gestation was terminated by cesarean section, in one case in the 29th week of gestation, and in the other case in the 30th week. The newborns showed no signs of lung hypoplasia and were successfully extubated on the 1st or 2nd day after delivery. Six months later the children did not exhibit any deviation from the normal development. Long-term amnioinfusion via a subcutaneously implanted port system could be used in humans with PPROM for prolongation of pregnancy and to avoid lung hypoplasia. Prospective randomized studies are ongoing. Copyright 2010 Elsevier Ireland Ltd. All rights reserved.

  12. Concentrations and speciation of polybrominated diphenyl ethers in human amniotic fluid

    Science.gov (United States)

    Miller, Mark F.; Chernyak, Sergei M.; Domino, Steven E.; Batterman, Stuart A.; Loch-Caruso, Rita

    2012-01-01

    Polybrominated diphenyl ethers (PBDEs) are persistent organic chemicals used as flame retardants in textiles, plastics, and consumer products. Although PBDE accumulation in humans has been noted since the 1970s, few studies have investigated PBDEs within the gestational compartment, and none to date has identified levels in amniotic fluid. The present study reports congener-specific brominated diphenyl ether (BDE) concentrations in second-trimester clinical amniotic fluid samples collected in 2009 from fifteen women in southeast Michigan, USA. Twenty-one BDE congeners were measured by GC/MS/NCI. The average total PBDE concentration was 3795 pg/ml amniotic fluid (range: 337 – 21842 pg/ml). BDE-47 and BDE-99 were identified in all samples. Based on median concentrations, the dominant congeners were BDE-208, 209, 203, 206, 207, and 47 representing 23, 16, 12, 10, 9 and 6%, respectively, of the total detected PBDEs. PBDE concentrations were identified in all amniotic fluid samples from southeast Michigan, supporting a need for further investigations of fetal exposure pathways and potential impacts on perinatal health. PMID:22236635

  13. ALPHA-FETOPROTEIN IN FETAL SERUM, AMNIOTIC-FLUID, AND MATERNAL SERUM

    NARCIS (Netherlands)

    VANLITH, JMM; BEEKHUIS, [No Value; VANLOON, AJ; MANTINGH, A; DEWOLF, BTHM; BREED, ASPM

    In order to gain more insight into the association between alpha-fetoprotein (AFP) and fetal chromosomal disorders, especially Down's syndrome, we measured AFP in fetal serum, amniotic fluid, and maternal serum at cordocentesis. We compared the concentration and gradient of AFP in these three

  14. Decreased alpha-fetoprotein in amniotic fluid and maternal serum in diabetic pregnancy

    DEFF Research Database (Denmark)

    Henriques, C U; Damm, P; Tabor, A

    1993-01-01

    OBJECTIVE: To determine a reference level for alpha-fetoprotein (AFP) in the amniotic fluid (AF) in pregnant women with insulin-dependent diabetes mellitus in order to suggest an explanation for the observed decrease in maternal serum AFP. METHODS: Alpha-fetoprotein was measured in AF, maternal...

  15. Amniotic fluid RNA gene expression profiling provides insights into the phenotype of Turner syndrome.

    Science.gov (United States)

    Massingham, Lauren J; Johnson, Kirby L; Scholl, Thomas M; Slonim, Donna K; Wick, Heather C; Bianchi, Diana W

    2014-09-01

    Turner syndrome is a sex chromosome aneuploidy with characteristic malformations. Amniotic fluid, a complex biological material, could contribute to the understanding of Turner syndrome pathogenesis. In this pilot study, global gene expression analysis of cell-free RNA in amniotic fluid supernatant was utilized to identify specific genes/organ systems that may play a role in Turner syndrome pathophysiology. Cell-free RNA from amniotic fluid of five mid-trimester Turner syndrome fetuses and five euploid female fetuses matched for gestational age was extracted, amplified, and hybridized onto Affymetrix(®) U133 Plus 2.0 arrays. Significantly differentially regulated genes were identified using paired t tests. Biological interpretation was performed using Ingenuity Pathway Analysis and BioGPS gene expression atlas. There were 470 statistically significantly differentially expressed genes identified. They were widely distributed across the genome. XIST was significantly down-regulated (p Turner syndrome transcriptome from other aneuploidies we previously studied. Manual curation of the differentially expressed gene list identified genes of possible pathologic significance, including NFATC3, IGFBP5, and LDLR. Transcriptomic differences in the amniotic fluid of Turner syndrome fetuses are due to genome-wide dysregulation. The hematologic/immune system differences may play a role in early-onset autoimmune dysfunction. Other genes identified with possible pathologic significance are associated with cardiac and skeletal systems, which are known to be affected in females with Turner syndrome. The discovery-driven approach described here may be useful in elucidating novel mechanisms of disease in Turner syndrome.

  16. Species-Specific Mechanisms of Neuron Subtype Specification Reveal Evolutionary Plasticity of Amniote Brain Development

    Directory of Open Access Journals (Sweden)

    Tadashi Nomura

    2018-03-01

    Full Text Available Summary: Highly ordered brain architectures in vertebrates consist of multiple neuron subtypes with specific neuronal connections. However, the origin of and evolutionary changes in neuron specification mechanisms remain unclear. Here, we report that regulatory mechanisms of neuron subtype specification are divergent in developing amniote brains. In the mammalian neocortex, the transcription factors (TFs Ctip2 and Satb2 are differentially expressed in layer-specific neurons. In contrast, these TFs are co-localized in reptilian and avian dorsal pallial neurons. Multi-potential progenitors that produce distinct neuronal subtypes commonly exist in the reptilian and avian dorsal pallium, whereas a cis-regulatory element of avian Ctip2 exhibits attenuated transcription suppressive activity. Furthermore, the neuronal subtypes distinguished by these TFs are not tightly associated with conserved neuronal connections among amniotes. Our findings reveal the evolutionary plasticity of regulatory gene functions that contribute to species differences in neuronal heterogeneity and connectivity in developing amniote brains. : Neuronal heterogeneity is essential for assembling intricate neuronal circuits. Nomura et al. find that species-specific transcriptional mechanisms underlie diversities of excitatory neuron subtypes in mammalian and non-mammalian brains. Species differences in neuronal subtypes and connections suggest functional plasticity of regulatory genes for neuronal specification during amniote brain evolution. Keywords: Ctip2, Satb2, multi-potential progenitors, transcriptional regulation, neuronal connectivity

  17. Targeted Expression of Stromelysin-1 in Mammary Gland Provides Evidence for a Role of Proteinases in Branching Morphogenesis and the Requirement for an Intact Basement Membrane for Tissue-specific Gene Expression

    Energy Technology Data Exchange (ETDEWEB)

    Sympson, Carolyn J; Talhouk, Rabih S; Alexander, Caroline M; Chin, Jennie R; Cliff, Shirley M; Bissell, Mina J; Werb, Zena

    1994-05-01

    The extracellular matrix (ECM) is an important regulator of the differentiated phenotype of mammary epithelial cells in culture. Despite the fact that ECM-degrading enzymes have been implicated in morphogenesis and tissue remodeling, there is little evidence for a direct role for such regulation in vivo. We generated transgenic mice that express autoactivated isoforms of the matrix metalloproteinase stromelysin-1, under the control of the whey acidic protein gene promoter, to examine the effect of inappropriate expression of this enzyme. Stromelysin-1 is implicated as the primary player in the loss of basement membrane and loss of function in the mammary gland during involution. The transgene was expressed at low levels in mammary glands of virgin female mice, leading to an unexpected phenotype: The primary ducts had supernumerary branches and showed precocious development of alveoli that expressed beta-casein at levels similar to that of an early- to mid-pregnant gland. Lactating glands showed high levels of transgene expression, with accumulation at the basement membrane, and a decrease in laminin and collagen IV, resulting in a loss of basement membrane integrity; this was accompanied by a dramatic alteration of alveolar morphology, with decreased size and shrunken lumina containing little beta-casein. During pregnancy, expression of endogenous whey acidic protein and beta-casein was reduced in transgenic glands, confirming the observed dependence of milk protein transcription of ECM in mammary epithelial cells in culture. These data provide direct evidence that stromelysin-1 activity can be morphogenic for mammary epithelial cells, inducing hyperproliferation and differentiation in virgin animals, and that its lytic activity can, indeed, disrupt membrane integrity and reduce mammary-specific function. We conclude that the balance of ECM-degrading enzymes with their inhibitors, and the associated regulation of ECM structure, is crucial for tissue-specific gene

  18. Nutritional intra-amniotic therapy increases survival in a rabbit model of fetal growth restriction

    Science.gov (United States)

    Illa, Miriam; Pla, Laura; Zamora, Monica; Crispi, Fatima; Gratacos, Eduard

    2018-01-01

    Objective To evaluate the perinatal effects of a prenatal therapy based on intra-amniotic nutritional supplementation in a rabbit model of intrauterine growth restriction (IUGR). Methods IUGR was surgically induced in pregnant rabbits at gestational day 25 by ligating 40–50% of uteroplacental vessels of each gestational sac. At the same time, modified-parenteral nutrition solution (containing glucose, amino acids and electrolytes) was injected into the amniotic sac of nearly half of the IUGR fetuses (IUGR-T group n = 106), whereas sham injections were performed in the rest of fetuses (IUGR group n = 118). A control group without IUGR induction but sham injection was also included (n = 115). Five days after the ligation procedure, a cesarean section was performed to evaluate fetal cardiac function, survival and birth weight. Results Survival was significantly improved in the IUGR fetuses that were treated with intra-amniotic nutritional supplementation as compared to non-treated IUGR animals (survival rate: controls 71% vs. IUGR 44% p = 0.003 and IUGR-T 63% vs. IUGR 44% p = 0.02), whereas, birth weight (controls mean 43g ± SD 9 vs. IUGR 36g ± SD 9 vs. IUGR-T 35g ± SD 8, p = 0.001) and fetal cardiac function were similar among the IUGR groups. Conclusion Intra-amniotic injection of a modified-parenteral nutrient solution appears to be a promising therapy for reducing mortality among IUGR. These results provide an opportunity to develop new intra-amniotic nutritional strategies to reach the fetus by bypassing the placental insufficiency. PMID:29466434

  19. Development of a novel method for amniotic fluid stem cell storage.

    Science.gov (United States)

    Zavatti, Manuela; Beretti, Francesca; Casciaro, Francesca; Comitini, Giuseppina; Franchi, Fabrizia; Barbieri, Veronica; Bertoni, Laura; De Pol, Anto; La Sala, Giovanni B; Maraldi, Tullia

    2017-08-01

    Current procedures for collection of human amniotic fluid stem cells (hAFSCs) indicate that cells cultured in a flask for 2 weeks can then be used for research. However, hAFSCs can be retrieved directly from a small amount of amniotic fluid that can be obtained at the time of diagnostic amniocentesis. The aim of this study was to determine whether direct freezing of amniotic fluid cells is able to maintain or improve the potential of a sub-population of stem cells. We compared the potential of the hAFSCs regarding timing of freezing, cells obtained directly from amniotic fluid aspiration (D samples) and cells cultured in a flask before freezing (C samples). Colony-forming-unit ability, proliferation, morphology, stemness-related marker expression, senescence, apoptosis and differentiation potential of C and D samples were compared. hAFSCs isolated from D samples expressed mesenchymal stem cells markers until later passages, had a good proliferation rate and exhibited differentiation capacity similar to hAFSCs of C samples. Interestingly, direct freezing induced a higher concentration of cells positive for pluripotency stem cell markers, without teratoma formation in vivo. This study suggests that minimal processing may be adequate for the banking of amniotic fluid cells, avoiding in vitro passages before the storage and exposure to high oxygen concentration, which affect stem cell properties. This technique might be a cost-effective and reasonable approach to the process of Good Manufacturing Process accreditation for stem-cell banks. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  20. Effect of trichloroethylene (TCE) toxicity on the enzymes of carbohydrate metabolism, brush border membrane and oxidative stress in kidney and other rat tissues.

    Science.gov (United States)

    Khan, Sheeba; Priyamvada, Shubha; Khan, Sara A; Khan, Wasim; Farooq, Neelam; Khan, Farah; Yusufi, A N K

    2009-07-01

    Trichloroethylene (TCE), an industrial solvent, is a major environmental contaminant. Histopathological examinations revealed that TCE caused liver and kidney toxicity and carcinogenicity. However, biochemical mechanism and tissue response to toxic insult are not completely elucidated. We hypothesized that TCE induces oxidative stress to various rat tissues and alters their metabolic functions. Male Wistar rats were given TCE (1000 mg/kg/day) in corn oil orally for 25 d. Blood and tissues were collected and analyzed for various biochemical and enzymatic parameters. TCE administration increased blood urea nitrogen, serum creatinine, cholesterol and alkaline phosphatase but decreased serum glucose, inorganic phosphate and phospholipids indicating kidney and liver toxicity. Activity of hexokinase, lactate dehydrogenase increased in the intestine and liver whereas decreased in renal tissues. Malate dehydrogenase and glucose-6-phosphatase and fructose-1, 6-bisphosphatase decreased in all tissues whereas increased in medulla. Glucose-6-phosphate dehydrogenase increased but NADP-malic enzyme decreased in all tissues except in medulla. The activity of BBM enzymes decreased but renal Na/Pi transport increased. Superoxide dismutase and catalase activities variably declined whereas lipid peroxidation significantly enhanced in all tissues. The present results indicate that TCE caused severe damage to kidney, intestine, liver and brain; altered carbohydrate metabolism and suppressed antioxidant defense system.

  1. [EXPERIMENTAL RESEARCH OF DIFFERENTIATION OF HUMAN AMNIOTIC MESENCHYMAL STEM CELLS INTO LIGAMENT CELLS IN VITRO].

    Science.gov (United States)

    Jin, Ying; Li, Yuwan; Zhang, Chenghao; Wu, Shuhong; Cheng, Daixiong; Liu, Yi

    2016-02-01

    To discuss whether human amniotic mesenchymal stem cells (hAMSCs) possesses the characteristic of mesenchymal stem cells, and could differentiate into ligament cells in vitro after induction. The hAMSCs were separated through enzyme digestion, and the phenotypic characteristics of hAMSCs were tested through flow cytometry. The cells at passage 3 were cultured with L-DMEM/F12 medium containing transforming growth factor beta1 (TGF-beta1) + basic fibroblast growth factor (bFGF) (group A), containing hyaluronic acid (HA) (group B), containing TGF-beta1+bFGF+HA (group C), and simple L-DMEM/F12 medium (group D) as control group. The morphology changes of cells in each group were observed by inverted phase contrast microscope at 21 days after induction; the cellular activities and proliferation were examined by sulforhodamine (SRB) colorimetric method; and specific mRNA and protein expressions of ligament including collagen type I, collagen type III, and tenascin C (TNC) were measured by real-time fluorescence quantitative PCR and immunohistochemical staining. The flow cytometry result indicated that hAMSCs expressed mesenchymal stem cell phenotype. After 21 days of induction, the cells in groups A, B, and C grew like spindle-shaped fibroblasts under inverted phase contrast microscope, and cells showed single shape, obvious directivity, and compact arrangement in group C. The SRB result indicated that the cells in each group reached the peak of growth curve at 6 days; the cellular activities of groups A, B, and C were significantly higher than that of group D at 6 days after induction. Also, the immunohistochemical staining results showed that no expressions of TNC were detected in 4 groups at 7 days; expressions of collagen type I in groups A, B, and C were significantly higher than that in group D at 7, 14, and 21 days (Pligament specific genes can be up-regulated and the synthesis of ligament specific proteins can be also strengthened. As a result, it can be used as

  2. Amnion and Chorion Membranes: Potential Stem Cell Reservoir with Wide Applications in Periodontics.

    Science.gov (United States)

    Gupta, Akanksha; Kedige, Suresh D; Jain, Kanu

    2015-01-01

    The periodontal therapy usually aims at elimination of disease causing bacteria and resolution of inflammation. It involves either resective or regenerative surgery to resolve the inflammation associated defects. Over the years, several methods have been used for achievement of periodontal regeneration. One of the oldest biomaterials used for scaffolds is the fetal membrane. The amniotic membranes of developing embryo, that is, amnion (innermost lining) and chorion (a layer next to it), have the properties with significant potential uses in dentistry. This paper reviews the properties, mechanism of action, and various applications of these placental membranes in general and specifically in Periodontics.

  3. STUDY OF BIOCHEMICAL PARAMETERS IN AMNIOTIC FLUID FOR ASSESSMENT OF FOETAL MATURITY IN CASES OF NORMAL PREGNANCY

    Directory of Open Access Journals (Sweden)

    Leela

    2015-11-01

    Full Text Available Assessment of foetal maturity had been proven of value in evaluating the foetal condition. Accurate assessment of foetal maturity is essential for the proper timing of delivery in various risk pregnancies. Amniotic Fluid analysis for foetal maturity had been of proven value. In the present study, study of biochemical parameters in amniotic fluid in respect of Creatinine, Uric Acid, Urea, Total Proteins, and Electrolytes i.e. Sodium, Potassium and Chloride has been done, along with Serum Electrolytes. Standard methodologies were adopted. The observations in the present study correlated with the works of Chadick et al and Pitkin and Zwirek. The levels of Creatinine, Uric Acid and Urea in Amniotic Fluid showed elevation, while Total Proteins and Serum Sodium showed a decline, as gestation progressed. The Serum and Amniotic Fluid Potassium and Chloride levels remain almost constant throughout the pregnancy. Thus, it is observed that the use of multiple parameters is desirable for accurate assessment of foetal maturity.

  4. Measurements of the cesium 137 concentration in surgical specimens of gynaecologic tumours and mother's milk, amniotic fluid, and in umbilical cords, 6 years after Chernobyl

    International Nuclear Information System (INIS)

    Czerwenka, K.F.; Heuss, F.; Teherani, D.K.

    1994-01-01

    In the present study, the cesium 137 content in various human tissues was examined 6 years after the Chernobyl reactor catastrophe. The measurements were performed with a gamma-ray spectrometer by means of a germanium/lithium detector. The median of cesium 137 was 20 mBq/ml in mother's milk. 60 mBq/ml in amniotic fluid, 105 mBq/g in umbilical cords. 51 mBq/g in ovarian tumours, and 140 mBq/g in mammary carcinomas. These values lay far below the permissible limit values of 528 mBq/ml or 528 mBq/g for persons not exposed to radiation. The problems of determining the upper limit were also discussed, and it was ascertained that despite the favorably low values recorded in this study a residual risk to health cannot be absolutely precluded in the light of present-day knowledge. (authors)

  5. Prenatal imaging of amniotic band sequence: utility and role of fetal MRI as an adjunct to prenatal US

    International Nuclear Information System (INIS)

    Neuman, Jeremy; Calvo-Garcia, Maria A.; Kline-Fath, Beth M.; Bitters, Constance; Merrow, Arnold C.; Guimaraes, Carolina V.A.; Lim, Foong-Yen

    2012-01-01

    Amniotic band sequence and its US manifestations have been well-described. There is little information, however, regarding the accuracy and utility of fetal MRI. To describe the MRI findings in amniotic band sequence and to compare the diagnostic accuracy of MRI and US. Prenatal MRI and US studies were retrospectively reviewed in 14 consecutive pregnancies with confirmed amniotic band sequence. Both studies were evaluated for amniotic band visualization, body part affected, type of deformity, umbilical cord involvement and vascular abnormality. Amniotic bands were confidently identified with MRI in 8 fetuses (57%), suggested with MRI in 3 fetuses (21%) and confidently seen by US in 13 fetuses (93%). Neither modality detected surgically proven bands on one fetus. Both techniques were equally able to define the body part affected and the type of deformity. At least one limb abnormality was visualized in all cases and truncal involvement was present in two cases. Cord involvement was identified in seven cases, with one case detected only by MRI. Fetal MRI is able to visualize amniotic bands and their secondary manifestations and could be complementary to prenatal US when fetal surgery is contemplated. (orig.)

  6. Role of amnioinfusion in the management of premature rupture of the membranes at <26 weeks' gestation.

    Science.gov (United States)

    Locatelli, A; Vergani, P; Di Pirro, G; Doria, V; Biffi, A; Ghidini, A

    2000-10-01

    We sought to evaluate whether serial amnioinfusions for persistent oligohydramnios can affect the perinatal and long-term outcomes in extreme cases of preterm premature rupture of membranes. All singleton pregnancies with preterm premature rupture of membranes at 4 days between January 1991 and June 1998 were included. Amniotic fluid volume was assessed as the maximum cord-free pocket with serial ultrasonographic examinations. Consenting women with persistent (>4 days) oligohydramnios (amniotic fluid amnioinfusions to maintain an amniotic fluid pocket >2 cm. The pregnancy, neonatal, and long-term neurologic outcomes of the cases that spontaneously maintained a median amniotic fluid pocket >2 cm (amnioinfusion-not-necessary group) were compared with those of women with oligohydramnios who underwent amnioinfusion but continued to have a median amniotic fluid pocket after preterm premature rupture of membranes amnioinfusion group). Statistical analysis included the Wilcoxon rank-sum test and the Fisher exact test, with a 2-tailed P amnioinfusions, which successfully restored a median amniotic fluid pocket >2 cm for >/=48 hours in 11 (30%) patients. This successful amnioinfusion group was comparable with the persistent oligohydramnios group (n = 25) in gestational age at first amnioinfusion (median, 20.2 weeks; range, 16-25.6 weeks; vs median, 20.3 weeks; range, 16.5-24.2 weeks; P =.4), number of amnioinfusions (median, 3; range, 1-9; vs median, 3; range, 1-5; P =.4), and interval between amnioinfusions (median, 6 days; range, 4-14 days; vs median, 8 days; range, 6-43 days; P =. 1). However, patients in the persistent oligohydramnios group had a significantly shorter interval to delivery, lower neonatal survival (20%), and higher rates of pulmonary hypoplasia (62%) and abnormal neurologic outcomes (60%) than the patients in the groups in which amnioinfusion was not necessary or was successful (all P amnioinfusions successfully alleviate oligohydramnios have a perinatal

  7. Thoraco-amniotic shunting for fetal pleural effusion--a case series.

    LENUS (Irish Health Repository)

    Walsh, J

    2011-11-15

    Fetal pleural effusion is a rare occurrence, with an incidence of 1 per 10-15,000 pregnancies. The prognosis is related to the underlying cause and is often poor. There is increasing evidence that in utero therapy with thoraco-amniotic shunting improves prognosis by allowing lung expansion thereby preventing hydrops and pulmonary hypoplasia. This is a review of all cases of fetal pleural effusion managed over an eight year period the National Maternity Hospital Dublin. Over the nine year period there were 21 cases of fetal pleural effusion giving an overall incidence of 1 per 9281 deliveries. Of these, 15 underwent thoraco-amniotic shunting. There were associated anomalies diagnosed in 5 (33%) of cases. The overall survival in our cohort was 53%. The presence of hydrops was a poor prognostic factor, with survival in cases with hydrops of 33% (3\\/9) compared to 83% (5\\/6) in those cases without associated hydrops.

  8. The Evolution and Fossil History of Sensory Perception in Amniote Vertebrates

    Science.gov (United States)

    Müller, Johannes; Bickelmann, Constanze; Sobral, Gabriela

    2018-05-01

    Sensory perception is of crucial importance for animals to interact with their biotic and abiotic environment. In amniotes, the clade including modern mammals (Synapsida), modern reptiles (Reptilia), and their fossil relatives, the evolution of sensory perception took place in a stepwise manner after amniotes appeared in the Carboniferous. Fossil evidence suggests that Paleozoic taxa had only a limited amount of sensory capacities relative to later forms, with the majority of more sophisticated types of sensing evolving during the Triassic and Jurassic. Alongside the evolution of improved sensory capacities, various types of social communication evolved across different groups. At present there is no definitive evidence for a relationship between sensory evolution and species diversification. It cannot be excluded, however, that selection for improved sensing was partially triggered by biotic interactions, e.g., in the context of niche competition, whereas ecospace expansion, especially during the Mesozoic, might also have played an important role.

  9. [Imbalance of system of glutamin - glutamic acid in the placenta and amniotic fluid at placental insufficiency].

    Science.gov (United States)

    Pogorelova, T N; Gunko, V O; Linde, V A

    2014-01-01

    Metabolism of glutamine and glutamic acid has been investigated in the placenta and amniotic fluid under conditions of placental insufficiency. The development of placental insufficiency is characterized by the increased content of glutamic acid and a decrease of glutamine in both placenta and amniotic fluid. These changes changes were accompanied by changes in the activity of enzymes involved in the metabolism of these amino acids. There was a decrease in glutamate dehydrogenase activity and an increase in glutaminase activity with the simultaneous decrease of glutamine synthetase activity. The compensatory decrease in the activity of glutamine keto acid aminotransferase did not prevent a decrease in the glutamine level. The impairments in the system glutamic acid-glutamine were more pronounced during the development of premature labor.

  10. Transplantation of human amniotic epithelial cells repairs brachial plexus injury:pathological and biomechanical analyses

    Institute of Scientific and Technical Information of China (English)

    Qi Yang; Min Luo; Peng Li; Hai Jin

    2014-01-01

    A brachial plexus injury model was established in rabbits by stretching the C6 nerve root. Imme-diately after the stretching, a suspension of human amniotic epithelial cells was injected into the injured brachial plexus. The results of tensile mechanical testing of the brachial plexus showed that the tensile elastic limit strain, elastic limit stress, maximum stress, and maximum strain of the injured brachial plexuses were signiifcantly increased at 24 weeks after the injection. The treat-ment clearly improved the pathological morphology of the injured brachial plexus nerve, as seen by hematoxylin eosin staining, and the functions of the rabbit forepaw were restored. These data indicate that the injection of human amniotic epithelial cells contributed to the repair of brachial plexus injury, and that this technique may transform into current clinical treatment strategies.

  11. Automated enzymatic measurement of lecithin, sphingomyelin, and phosphatidylglycerol in amniotic fluid.

    Science.gov (United States)

    Bradley, C A; Salhany, K E; Entman, S S; Aleshire, S L; Parl, F F

    1987-01-01

    We describe methods for automated enzymatic measurement of lecithin, sphingomyelin, and phosphatidylglycerol in amniotic fluid. Phospholipase C (EC 3.1.4.3) and sphingomyelin phosphodiesterase (EC 3.1.4.12) are reacted with lecithin and sphingomyelin, respectively, to liberate phosphocholine. Phosphocholine is then reacted with alkaline phosphatase, choline oxidase, peroxidase, and 4-aminoantipyrine to form a colored complex, for which the absorbance at 500 nm is measured with a centrifugal analyzer. Phosphatidylglycerol is hydrolyzed by phospholipase D (EC 3.1.4.4) to form glycerol, which is subsequently reacted with ATP and NAD+ in the presence of glycerol kinase and glycerol-3-phosphate dehydrogenase to yield NADH. The absorbance of the NADH formed is measured at 340 nm. These methods provide a simple, rapid, and accurate alternative to thin-layer chromatography for determination of phospholipids in amniotic fluid for assessment of fetal lung maturity.

  12. Amniotic fluid and colostrum as potential diets in the critical care of preterm infants

    DEFF Research Database (Denmark)

    Støy, Ann Cathrine Findal; Viberg Østergaard, Mette; Torp Sangild, Per

    2015-01-01

    Amniotic fluid is the enteral “diet” of the developing fetus, while the first mammary gland secretion, colostrum, is the natural diet of the newborn mammal. Both diets contain nutrients but also growth factors, immune-modulating components, and antibacterial agents that support perinatal organ...... development, particularly of the gastrointestinal (GI) tract. Birth requires a sudden transition to nutrient uptake via the GI tract and exposure to microorganisms. Ingestion of amniotic fluid before birth and of colostrum just after birth helps to adapt GI functions and provides protection against...... and colostrum, and we describe how these fluids may have a therapeutic potential for GI conditions in some pediatric patients, particularly preterm infants. The composition of the two fluids varies widely among different species and the effects are likely highly species specific. Some effects may however...

  13. Management of interstitial ectopic pregnancies with a combined intra-amniotic and systemic approach.

    Science.gov (United States)

    Swank, Morgan L; Harken, Tabetha R; Porto, Manuel

    2013-08-01

    Approximately 2% of all pregnancies are ectopic; of these, 4% are interstitial or cervical. There exists no clear consensus as to whether surgical or medical management is superior. We present three cases of advanced nonfallopian tube ectopic pregnancies from 6 to 8 weeks of gestation. Our first two cases were managed with a combined intrafetal, intra-amniotic and systemic approach using methotrexate and potassium chloride, whereas our third case was managed with an intra-amniotic approach alone. Our combined approach cases were successful, with resolution of human chorionic gonadotropin in 50 and 34 days, whereas our single approach case re-presented with bleeding requiring uterine artery embolization and operative removal of products of conception. Patients presenting with advanced interstitial or cervical pregnancies who are clinically stable can be offered medical management with a combined approach.

  14. Membrane fusion

    DEFF Research Database (Denmark)

    Bendix, Pól Martin

    2015-01-01

    At Stanford University, Boxer lab, I worked on membrane fusion of small unilamellar lipid vesicles to flat membranes tethered to glass surfaces. This geometry closely resembles biological systems in which liposomes fuse to plasma membranes. The fusion mechanism was studied using DNA zippering...... between complementary strands linked to the two apposing membranes closely mimicking the zippering mechanism of SNARE fusion complexes....

  15. Aging of intrauterine tissues in spontaneous preterm birth and preterm premature rupture of the membranes: A systematic review of the literature.

    Science.gov (United States)

    Polettini, J; Dutta, E H; Behnia, F; Saade, G R; Torloni, M R; Menon, R

    2015-09-01

    Many adverse pregnancy outcomes (APOs), including spontaneous preterm birth (PTB), are associated with placental dysfunction. Recent clinical and experimental evidences suggest that premature aging of the placenta may be involved in these events. Although placental aging is a well-known concept, the mechanisms of aging during normal pregnancy and premature aging in APOs are still unclear. This review was conducted to assess the knowledge on placental aging related biochemical changes leading to placental dysfunction in PTB and/or preterm premature rupture of membranes (pPROM). We performed a systematic review of studies published over the last 50 years in two electronic databases (Pubmed and Embase) on placental aging and PTB or pPROM. The search yielded 554 citations, 30 relevant studies were selected for full-text review and three were included in the review. Only one study reported oxidative stress-related aging and degenerative changes in human placental membranes and telomere length reduction in fetal cells as part of PTB and/or pPROM mechanisms. Similarly, two animal studies reported findings of decidual senescence and referred to PTB mechanisms. Placental and fetal membrane oxidative damage and telomere reduction are linked to premature aging in PTB and pPROM but the risk factors and biomolecular pathways causing this phenomenon are not established in the literature. However, no biomarkers or clinical indicators of premature aging as a pathology of PTB and pPROM have been reported. We document major knowledge gaps and propose several areas for future research to improve our understanding of premature aging linked to placental dysfunction. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Tritium-labelled hemicholinium-3 ([3H]HC-3): membrane binding properties and potential uses for a novel presynaptic marker in cholinergically-innervated tissues

    International Nuclear Information System (INIS)

    Vickroy, T.W.; Watson, M.; Roeske, W.R.; Yamamura, H.I.

    1986-01-01

    Sodium-dependent high-affintiy choline uptake (SDHACU) is the primary regulatory step in acetylcholine biosynthesis and subserves an essential function in cholinergically-mediated neurotransmission. Recent studies with [ 3 H]hemicholinium-3 ([ 3 H]HC-3), a potent competitive inhibitor of SDHACU, reveal that closely associated membrane sites mediate high-affinity [ 3 H]HC-3 binding and SDHACU. In this report, supportive evidences for this association are presented and potential uses of [ 3 H]HC-3 are outlined for studies of disorders that involve cholinergic nervous system dysfunction. 40 refs.; 1 figure

  17. DNA methylation patterns of imprinting centers for H19, SNRPN, and KCNQ1OT1 in single-cell clones of human amniotic fluid mesenchymal stem cell

    Directory of Open Access Journals (Sweden)

    Hsiu-Huei Peng

    2012-09-01

    Conclusion: In conclusion, human amniotic fluid mesenchymal stem cells contain a unique epigenetic signature during in vitro cell culture. H19 and KCNQ1OT1 possessed a substantial degree of hypermethylation status, and variable DNA methylation patterns of SNRPN was observed during in vitro cell culture of human amniotic fluid mesenchymal stem cells. Our results urge further understanding of epigenetic status of human amniotic fluid mesenchymal stem cells before it is applied in cell replacement therapy.

  18. Metabolite Profiling of Human Amniotic Fluid by Hyphenated Nuclear Magnetic Resonance Spectroscopy

    OpenAIRE

    Graça, Gonçalo; Duarte, Iola F.; Goodfellow, Brian J.; Carreira, Isabel M.; Couceiro, Ana Bela; Domingues, Maria do Rosário; Spraul, Manfred; Tseng, Li-Hong; Gil, Ana M.

    2008-01-01

    The metabolic profiling of human amniotic fluid (HAF) is of potential interest for the diagnosis of disorders in the mother or the fetus. In order to build a comprehensive metabolite database for HAF, hyphenated NMR has been used, for the first time, for systematic HAF profiling. Experiments were carried out using reverse-phase (RP) and ion-exchange liquid chromatography (LC), in order to detect less and more polar compounds, respectively. RP-LC conditions achieved good separation of amino ac...

  19. Accuracy of real-time polymerase chain reaction for Toxoplasma gondii in amniotic fluid.

    Science.gov (United States)

    Wallon, Martine; Franck, Jacqueline; Thulliez, Philippe; Huissoud, Cyril; Peyron, François; Garcia-Meric, Patricia; Kieffer, François

    2010-04-01

    To provide clinicians with information about the accuracy of real-time polymerase chain reaction (PCR) analysis of amniotic fluid for the prenatal diagnosis of congenital Toxoplasma infection. This was a prospective cohort study of women with Toxoplasma infection identified by prenatal screening in three centers routinely carrying out real-time PCR for the detection of Toxoplasma gondii in amniotic fluid. The data available were gestational age at maternal infection, types and dates of maternal treatment, results of amniocentesis and neonatal work-up and definitive infectious status of the child. We estimated sensitivity, specificity and positive and negative predictive values both overall and per trimester of pregnancy at the time of maternal infection. Polymerase chain reaction analysis was carried out on amniotic fluid for 261 of the 377 patients included (69%). It was accurate with the exception of four negative results in children who were infected. Overall sensitivity and negative predictive value were 92.2% (95% confidence interval [CI] 81-98%) and 98.1% (95% CI 95-99.5%), respectively. There was no significant association with the trimester of pregnancy during which maternal infection occurred. Specificity and positive predictive values of 100% were obtained for all trimesters. Real-time PCR analysis significantly improves the detection of T. gondii on amniotic fluid. It provides an accurate tool to predict fetal infection and to decide on appropriate treatment and surveillance. However, postnatal follow-up remains necessary in the first year of life to fully exclude infection in children for whom PCR results were negative. III.

  20. Characteristics of human amniotic fluid mesenchymal stem cells and their tropism to human ovarian cancer.

    Directory of Open Access Journals (Sweden)

    Liru Li

    Full Text Available The mesenchymal stem cells (MSCs derived from amniotic fluid (AF have become an attractive stem cells source for cell-based therapy because they can be harvested at low cost and avoid ethical disputes. In human research, stem cells derived from AF gradually became a hot research direction for disease treatment, specifically for their plasticity, their reduced immunogenicity and their tumor tropism regardless of the tumor size, location and source. Our work aimed to obtain and characterize human amniotic fluid mesenchymal stem cells (AFMSCs and detect their ovarian cancer tropsim in nude mice model. Ten milliliters of twenty independent amniotic fluid samples were collected from 16-20 week pregnant women who underwent amniocentesis for fetal genetic determination in routine prenatal diagnosis in the first affiliated hospital of Harbin medical university. We successfully isolated the AFMSCs from thirteen of twenty amniotic fluid samples. AFMSCs presented a fibroblastic-like morphology during the culture. Flow cytometry analyses showed that the cells were positive for specific stem cell markers CD73,CD90, CD105, CD166 and HLA-ABC (MHC class I, but negative for CD 45,CD40, CD34, CD14 and HLA-DR (MHC class II. RT-PCR results showed that the AFMSCs expressed stem cell marker OCT4. AFMSCs could differentiate into bone cells, fat cells and chondrocytes under certain conditions. AFMSCs had the high motility to migrate to ovarian cancer site but didn't have the tumorigenicity. This study enhances the possibility of AFMSCs as drug carrier in human cell-based therapy. Meanwhile, the research emphasis in the future can also put in targeting therapy of ovarian cancer.

  1. Prophylactic cefazolin in amnioinfusions administered for meconium-stained amniotic fluid.

    Science.gov (United States)

    Edwards, R K; Duff, P

    1999-01-01

    OBJECTIVE: To determine if amnioinfusion with an antibiotic solution decreased the rate of clinical chorioamnionitis and puerperal endometritis in patients with meconium-stained amniotic fluid. METHODS: Patients in labor at 36 weeks of gestation or greater with singleton pregnancies and meconium-stained amniotic fluid were randomized to receive either cefazolin, 1 g/1,000 mL, of normal saline (n = 90) or normal saline (n = 93) amnioinfusion. Rates of clinically diagnosed chorioamnionitis and endometritis and of suspected and culture-proven neonatal infection were determined. RESULTS: Between the study and control groups, the incidences of clinical chorioamnionitis (7.8% vs. 8.6%), endometritis (2.4% vs. 3.5%), aggregate intrauterine infection (10.0% vs. 11.8%), suspected neonatal infection (17.8% vs. 21.5%), and proven neonatal infection (0.0% vs. 2.2%) were not significantly different. CONCLUSIONS: Prophylactic use of cefazolin in amnioinfusions did not significantly reduce rates of maternal or neonatal infection in patients with meconium-stained amniotic fluid. PMID:10371474

  2. Effect of amnioinfusion for meconium stained amniotic fluid on perinatal outcome.

    Science.gov (United States)

    Ashfaq, F; Shah, A A

    2004-06-01

    To see the effect of amnioinfusion on perinatal outcome in cases of meconium staining of liquor. This study was conducted in department of Obstetrics and Gynaecology, unit 1, Jinnah Postgraduate Medical Centre, Karachi, from 1st January 1998 to 31st December 2000. Four hundred patients were included in this study, assigning 200 for amnioinfusion and 200 as control. All patients were matched in both the groups with respect to age, antenatal booking, parity, gestational age, stage of labour, colour of amniotic fluid and fetal birth weight. Both the groups were found to be comparable. The rate of Caesarean section was found to be 37% in amnioinfusion group, which collaborates with other international studies. The fetal outcome was better i.e. 91% alive and healthy, after amnioinfusion due to dilution of meconium stained amniotic fluid with physiological solutions. The perinatal outcome was recorded by Apgar score at 5 minutes. The perinatal morbidity and mortality both were significantly lowered and was found to be 6% as compared to 14% in control, which was also noticed by less number of admissions in nursery i.e. 12% and perinatal deaths. The incidence of meconium aspiration syndrome was found to be 56% in control and was reduced to 22% after amnioinfusion in the other arm of the study. These results are very encouraging and suggestion can be safely made that in future amnioinfusion will be the ideal method of preventing fetal distress due to meconium stained amniotic fluid.

  3. Role of intrapartum transcervical amnioinfusion in patients with meconium-stained amniotic fluid.

    Science.gov (United States)

    Bhatia, Pushpa; Reena, Kumari; Nangia, Sangita

    2013-03-01

    The study was undertaken to evaluate maternal, perinatal outcomes following transcervical intrapartum amnioinfusion in women with meconium-stained amniotic fluid. A prospective comparative study was conducted on 100 women with meconium-stained amniotic fluid in labor. Group A: study group (50 cases) received amnioinfusion. Group B: control group (50 cases) did not receive amnioinfusion. FHR monitoring was done using cardiotocography. Significant relief from variable decelerations was seen in 68.18 % cases in the amnioinfusion group as compared to 7.1 % cases in the control group. 78 % cases who were given amnioinfusion had vaginal delivery as compared to 18 % cases in the control group. Fourteen percent cases in the study group had cesarean delivery as compared to 68 % cases in the control group. Meconium aspiration syndrome was seen in six percent neonates in the study group as compared to 20 % in the control group. Two neonates died in the control group due to meconium aspiration syndrome. There was no maternal mortality or major maternal complication. Intrapartum transcervical amnioinfusion is valuable in patients with meconium-stained amniotic fluid.

  4. Maternal Plasma and Amniotic Fluid Chemokines Screening in Fetal Down Syndrome

    Directory of Open Access Journals (Sweden)

    Piotr Laudanski

    2014-01-01

    Full Text Available Objective. Chemokines exert different inflammatory responses which can potentially be related to certain fetal chromosomal abnormalities. The aim of the study was to determine the concentration of selected chemokines in plasma and amniotic fluid of women with fetal Down syndrome. Method. Out of 171 amniocentesis, we had 7 patients with confirmed fetal Down syndrome (15th–18th weeks of gestation. For the purpose of our control, we chose 14 women without confirmed chromosomal aberration. To assess the concentration of chemokines in the blood plasma and amniotic fluid, we used a protein macroarray, which allows the simultaneous determination of 40 chemokines per sample. Results. We showed significant decrease in the concentration of 4 chemokines, HCC-4, IL-28A, IL-31, and MCP-2, and increase in the concentration of CXCL7 (NAP-2 in plasma of women with fetal Down syndrome. Furthermore, we showed decrease in concentration of 3 chemokines, ITAC, MCP-3, MIF, and increase in concentration of 4 chemokines, IP-10, MPIF-1, CXCL7, and 6Ckine, in amniotic fluid of women with fetal Down syndrome. Conclusion. On the basis of our findings, our hypothesis is that the chemokines may play role in the pathogenesis of Down syndrome. Defining their potential as biochemical markers of Down syndrome requires further investigation on larger group of patients.

  5. Evaluation of two real time PCR assays for the detection of bacterial DNA in amniotic fluid.

    Science.gov (United States)

    Girón de Velasco-Sada, Patricia; Falces-Romero, Iker; Quiles-Melero, Inmaculada; García-Perea, Adela; Mingorance, Jesús

    2018-01-01

    The aim of this study was to evaluate two non-commercial Real-Time PCR assays for the detection of microorganisms in amniotic fluid followed by identification by pyrosequencing. We collected 126 amniotic fluids from 2010 to 2015 for the evaluation of two Real-Time PCR assays for detection of bacterial DNA in amniotic fluid (16S Universal PCR and Ureaplasma spp. specific PCR). The method was developed in the Department of Microbiology of the University Hospital La Paz. Thirty-seven samples (29.3%) were positive by PCR/pyrosequencing and/or culture, 4 of them were mixed cultures with Ureaplasma urealyticum. The Universal 16S Real-Time PCR was compared with the standard culture (81.8% sensitivity, 97.4% specificity, 75% positive predictive value, 98% negative predictive value). The Ureaplasma spp. specific Real-Time PCR was compared with the Ureaplasma/Mycoplasma specific culture (92.3% sensitivity, 89.4% specificity, 50% positive predictive value, 99% negative predictive value) with statistically significant difference (p=0.005). Ureaplasma spp. PCR shows a rapid response time (5h from DNA extraction until pyrosequencing) when comparing with culture (48h). So, the response time of bacteriological diagnosis in suspected chorioamnionitis is reduced. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Amniotic fluid volume: Rapid MR-based assessment at 28-32 weeks gestation

    Energy Technology Data Exchange (ETDEWEB)

    Hilliard, N.J.; Hawkes, R.; Patterson, A.J.; Graves, M.J.; Priest, A.N.; Hunter, S.; Set, P.A.; Lomas, D.J. [Cambridge University Hospitals NHS Foundation Trust, Department of Radiology, Cambridge (United Kingdom); Lees, C. [Imperial College Healthcare NHS Trust, Department of Obstetrics and Fetal Medicine, London (United Kingdom)

    2016-10-15

    This work evaluates rapid magnetic resonance projection hydrography (PH) based amniotic fluid volume (AFV) estimates against established routine ultrasound single deepest vertical pocket (SDVP) and amniotic fluid index (AFI) measurements, in utero at 28-32 weeks gestation. Manual multi-section planimetry (MSP) based measurement of AFV is used as a proxy reference standard. Thirty-five women with a healthy singleton pregnancy (20-41 years) attending routine antenatal ultrasound were recruited. SDVP and AFI were measured using ultrasound, with same day MRI assessing AFV with PH and MSP. The relationships between the respective techniques were assessed using linear regression analysis and Bland-Altman method comparison statistics. When comparing estimated AFV, a highly significant relationship was observed between PH and the reference standard MSP (R{sup 2} = 0.802, p < 0.001). For the US measurements, SDVP measurement related most closely to amniotic fluid volume, (R{sup 2} = 0.470, p < 0.001), with AFI demonstrating a weaker relationship (R{sup 2} = 0.208, p = 0.007). This study shows that rapid MRI based PH measurement is a better predictor of AFV, relating more closely to our proxy standard than established US techniques. Although larger validation studies across a range of gestational ages are required this approach could form part of MR fetal assessment, particularly where poly- or oligohydramnios is suspected. (orig.)

  7. A microfluidic device for separation of amniotic fluid mesenchymal stem cells utilizing louver-array structures.

    Science.gov (United States)

    Wu, Huei-Wen; Lin, Xi-Zhang; Hwang, Shiaw-Min; Lee, Gwo-Bin

    2009-12-01

    Human mesenchymal stem cells can differentiate into multiple lineages for cell therapy and, therefore, have attracted considerable research interest recently. This study pr