WorldWideScience
1

Establishment, characterization and immortalization of a fibroblast cell line from the Chinese red belly toad Bombina maxima skin  

OpenAIRE

The skin of the amphibian Bombina maxima is rich in biologically active proteins and peptides, most of which have mammalian analogues. The physiological functions of most of the mammalian analogues are still unknown. Thus, Bombina maxima skin may be a promising model to reveal the physiological role of these proteins and peptides because of their large capacity for secretion. To investigate the physiological role of these proteins and peptides in vitro, a fibroblast cell line was successfully...

Xiang, Yang; Gao, Qian; Su, Weiting; Zeng, Lin; Wang, Jinhuan; Hu, Yi; Nie, Wenhui; Ma, Xutong; Zhang, Yong; Lee, Wenhui; Zhang, Yun

2011-01-01

2

Critical role of GSH in Sulfur Mustard-induced Oxidative Stress and Cytotoxicity in Human Skin Fibroblast Cell Line  

OpenAIRE

In this study the role of glutathione (GSH) in sulfur mustard -induced oxidative stress and cytotoxicity, in human skin fibroblast cell line (HF2FF) was evaluated. Sulfur mustard-induced superoxide radical and hydrogen peroxide formation were evaluated by determination of superoxide dismutase and catalase activity in cell lysate. The cytotoxicity of sulfur mustard was estimated by lactate dehydrogenase leakage. The intracellular GSH content was modulated by N-acetylcysteine (NAC), a GSH precu...

Ali Beman Zaree Mahmoudabad; Mehdy Saberi; Jilla Pirzad

2008-01-01

3

Critical role of GSH in Sulfur Mustard-induced Oxidative Stress and Cytotoxicity in Human Skin Fibroblast Cell Line  

Directory of Open Access Journals (Sweden)

Full Text Available In this study the role of glutathione (GSH in sulfur mustard -induced oxidative stress and cytotoxicity, in human skin fibroblast cell line (HF2FF was evaluated. Sulfur mustard-induced superoxide radical and hydrogen peroxide formation were evaluated by determination of superoxide dismutase and catalase activity in cell lysate. The cytotoxicity of sulfur mustard was estimated by lactate dehydrogenase leakage. The intracellular GSH content was modulated by N-acetylcysteine (NAC, a GSH precursor, and buthionine sulfoximine (BSO, a specific GSH synthesis inhibitor. It was found that sulfur mustard exposure led to a dose-and time-dependent decrease in GSH content in HF2FF cells. NAC increased intracellular GSH level and protected the cells against sulfur mustard-induced reactive oxygen species formation and lactate dehydrogenase leakage. In contrast, buthionine sulfoximine pretreatment depleted cellular GSH and enhanced the susceptibility of HF2FF to the cytotoxic effects of sulfur mustard. These results indicated that GSH plays a critical role in protecting HF2FF cell line against sulfur mustar-induced cell injury, most probably through its antioxidant activity.

Ali Beman Zaree Mahmoudabad

2008-01-01

4

Lymphoblastoid lines and skin fibroblasts from patients with tuberous sclerosis are abnormally sensitive to ionizing radiation and to a radiomimetic chemical  

International Nuclear Information System (INIS)

Lymphoblastoid lines, derived by transforming peripheral blood lymphocytes with Epstein-Barr virus, and skin fibroblast lines were established from two patients with tuberous sclerosis. The number of viable lymphoblastoid cells was determined by their ability to exclude the vital dye trypan blue after their irradiation with x-rays or 254 nm ultraviolet light. The growth of fibroblasts was determined by their ability to form colonies after treatment with the radiomimetic, DNA-damaging chemical N-methyl-N'-nitro-N-nitrosoguanidine. The tuberous sclerosis lymphoblastoid lines were hypersensitive to x-rays but had normal sensitivity to the ultraviolet radiation. The tuberous sclerosis fibroblast lines were hypersensitive to the N-methyl-N'-nitro-N-nitrosoguanidine. The hypersensitivity of tuberous sclerosis cells to x-rays and to N-methyl-N'-nitro-N-nitrosoguanidine is believed to reflect defective repair of DNA damaged by these agents and may provide the basis for in vitro, including prenatal, diagnostic tests for tuberous sclerosis

5

Cytoskeletal proteins from human skin fibroblasts, peripheral blood leukocytes, and a lymphoblastoid cell line compared by two-dimensional gel electrophoresis  

International Nuclear Information System (INIS)

Differences in proteins between cells grown as suspension cultures and those grown as attached cultures were studied by comparing the proteins of detergent-resistant cytoskeletons prepared from peripheral blood leukocytes and a lymphoblastoid cell line (GM607) (both grown as suspension cultures) and those of human skin fibroblasts (grown as attached cultures) by two-dimensional gel electrophoresis. The major cytoskeletal proteins of the leukocytes were also present in the protein pattern of GM607 cytoskeletons. In contrast, the fibroblast cytoskeletal protein pattern contained four groups of proteins that differed from the patterns of the leukocytes and GM607. In addition, surface labeling of GM607 and human fibroblasts with 125I demonstrated that substantial amounts of vimentin and actin are exposed at the surface of the attached fibroblasts, but there is little evidence of similar exposure at the surface of the suspension-grown GM607. These results demonstrate some differences in cytoskeletal protein composition between different types of cells could be related to their ability or lack of ability to grow as attached cells in tissue culture

6

Power line frequency electromagnetic fields do not increase the rate of protein synthesis in human skin fibroblasts as previously reported.  

Science.gov (United States)

Rodemann et al. [Rodemann et al. (1987): Biochem Biophys Res Commun 145:1-9] reported that human skin fibroblasts increase their rate of protein synthesis by as much as over ninefold in response to long term exposure to 20 Hz, 8.4 mT (84 G) magnetic fields. Here we report studies of protein synthesis using an identical cell type, exposure conditions, and the same means of measuring protein synthetic rates. Our initial goal was to determine if the earlier results could be replicated, but we found an inconsistency in the earlier protocol. It exposed cells to [(3)H]leucine for 6 h prior to measuring incorporation into protein. We found, however, that 24 h is required for [(3)H]leucine to reach a steady state distribution across the cells' plasma membranes. In addition, we typically measured 100-200 cpm/thousand cells. This is four- to eightfold higher than the 19-28 cpm/1000 cells previously reported. Using these conditions, we could find no significant difference in protein synthesis rates between control cells and cells exposed for up to three weeks in an identical electromagnetic field. In addition, we investigated the effects of a 60 Hz field since that is the frequency used for electric power distribution in the United States. Again, we could find no significant effect of this field on rates of protein synthesis, even after 21 days of exposure. PMID:12955751

Shi, Biao; Isseroff, R Rivkah; Nuccitelli, Richard

2003-10-01

7

Estrogen Protection in Friedreich's Ataxia Skin Fibroblasts  

OpenAIRE

Estrogens have been shown to have protective effects on a wide range of cell types and animal models for many neurodegenerative diseases. The present study demonstrates the cytoprotective effects of 17?-estradiol (E2) and estrogen-like compounds in an in vitro model of Friedreich's ataxia (FRDA) using human donor FRDA skin fibroblasts. FRDA fibroblasts are extremely sensitive to free radical damage and oxidative stress, produced here using l-buthionine (S,R)-sulfoximine to inhibit de novo gl...

Richardson, Timothy E.; Yang, Shao-hua; Wen, Yi; Simpkins, James W.

2011-01-01

8

Alteration of Skin Properties with Autologous Dermal Fibroblasts  

Directory of Open Access Journals (Sweden)

Full Text Available Dermal fibroblasts are mesenchymal cells found between the skin epidermis and subcutaneous tissue. They are primarily responsible for synthesizing collagen and glycosaminoglycans; components of extracellular matrix supporting the structural integrity of the skin. Dermal fibroblasts play a pivotal role in cutaneous wound healing and skin repair. Preclinical studies suggest wider applications of dermal fibroblasts ranging from skin based indications to non-skin tissue regeneration in tendon repair. One clinical application for autologous dermal fibroblasts has been approved by the Food and Drug Administration (FDA while others are in preclinical development or various stages of regulatory approval. In this context, we outline the role of fibroblasts in wound healing and discuss recent advances and the current development pipeline for cellular therapies using autologous dermal fibroblasts. The microanatomic and phenotypic differences of fibroblasts occupying particular locations within the skin are reviewed, emphasizing the therapeutic relevance of attributes exhibited by subpopulations of fibroblasts. Special focus is provided to fibroblast characteristics that define regional differences in skin, including the thick and hairless skin of the palms and soles as compared to hair-bearing skin. This regional specificity and functional identity of fibroblasts provides another platform for developing regional skin applications such as the induction of hair follicles in bald scalp or alteration of the phenotype of stump skin in amputees to better support their prosthetic devices.

Rajesh L. Thangapazham

2014-05-01

9

Establishment and cryopreservation of a skin fibroblast cell line derived from Yunnan semi-fine wool sheep in the presence of synthetic ice blocker.  

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In this study, the fibroblasts cell line derived from ear marginal tissue of Yunnan semi-fine wool sheep was successfully established using the primary explants technique and cryopreservation technology. Additionally, the protective effect of synthetic ice blocker (SIB) including 1, 3-cyclohexanediol (1, 3-CHD) and 1, 4-cyclohexanediol (1, 4-CHD) on frozen fibroblast cells was also assessed and compared. Propidium iodide (PI) was used to stain the dead cells following cryopreservation and thawing. The results showed that compared with Medium 199 (M199) and Dulbecco's modified Eagle's medium : Nutrient Mixture F-12 (1 : 1) Mixture (DMEM/F12), Dulbecco's modified Eagle's medium (DMEM) may be more suitable for the primary culture of fibroblast cells of Yunnan semi-fine wool sheep. The growth curve of cells is a typical "S" type. After subculture for four days, the cells entered the plateau phase and began to degenerate. Biological analysis showed that the population doubling time (PDT) for subculturing fibroblast cells was approximately 26h. The Karyotyping data indicated that the percentage of fibroblast cells with normal chromosome number 2n = 54 was over 90% following subculture for 10 passages. Moreover, the tests for bacteria, fungi, viruses and mycoplasma were negative. After serial subculture for 5 generations, the fibroblast cells were cryopreserved in the presence or absence of 1, 3-CHD or 1, 4-CHD. The data indicated that with increase of the synthetic ice blocker concentrations, the viability of frozen-thawed fibroblast cells was firstly increased and then decreased. When the concentration of 1, 3-CHD or 1, 4-CHD was 50 mM, the viable percentage of frozen-thawed fibroblast cells was 91.93% +/- 2.24% and 94.13% +/- 0.55% respectively and significantly higher than that of the cells frozen in the absence of synthetic ice blockers (88.10% +/- 1.49%, P cell line of Yunnan semi-fine wool sheep was firstly established in this study. Additionally, the presence of synthetic ice blocker can increase the viability of frozen-thawed sheep fibroblast cell line. PMID:24448770

Wu, Shuai Shuai; Li, Dong Jiang; Lv, Chun Rong; Yang, Hong Yuan; Zhu, Lan; Li, Wei Juan; Quan, Guo Bo; Hong, Qiong Hua

2013-01-01

10

Protective and restorative effects of a Commiphora mukul gum resin and triheptanoin preparation on the CCL-110 skin fibroblast cell line.  

Science.gov (United States)

Coenzyme Q10 (CoQ10) is a major ingredient in skin care products because of its anti-wrinkle effects, although it has some side effects especially at higher amounts. In this study, we compare the anti-wrinkle related properties of CoQ10 and a proprietary Commiphora mukul gum resin (guggul) and triheptanoin preparation (GU-TC7). GU-TC7 is prepared with a supercritical CO?-co-solvent extraction with ethanol, standardized to 2% guggulsterones and triheptanoin, a triglyceride composed of three 7-carbon fatty acids. Treatment of CCL-110 skin fibroblasts with GU-TC7 demonstrates a mild proliferative effect compared to CoQ10 and increased type I collagen synthesis. Additionally, GU-TC7 inhibited matrix metalloproteinase-1 (MMP-1) expression in a dose-dependent manner at 20-100??g?mL?¹ and inhibited human elastase expression by more than 50% as compared to no elastase inhibition with CoQ10 treatment. These results suggest that GU-TC7 possesses properties that are applicable to the treatment of wrinkles and may be considered for its further evaluation in skin care products. PMID:22084831

Ramachandran, C; Quirin, K-W; Resek, A; Melnick, S J

2012-04-01

11

Induction of growth stimulation in skin fibroblasts from retinoblastoma donors after ionizing radiation  

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Skin fibroblasts from normal children and two children with a 13q14 deletion retinoblastoma (Rb) were submitted to fractionated doses of ..gamma.. radiations. Irradiation reduced the population doublings in normal fibroblasts and the decline was inversely related to the dose. An increase in population doublings was obtained with one of the Rb cell lines. Foci appeared in the irradiated culture of the other Rb donor. It is suggested that fibroblasts from patients with Rb are able to express some phenotypical properties of transformed cells, perhaps related to factors rendering them more susceptible to carcinogens.

Diatloff-Zito, C.; Macieira-Coelho, A. (Institut de Cancerologie et d' Immunogenetique, I.N.S.E.R.M. U. 50, 94 - Villejuif (France)); Turleau, C.; Cabanis, M.O.; Grouchy, J. de (Hopital Necker et des Enfants-Malades, 75 - Paris (France))

1983-11-07

12

Induction of growth stimulation in skin fibroblasts from retinoblastoma donors after ionizing radiation  

International Nuclear Information System (INIS)

Skin fibroblasts from normal children and two children with a 13q14 deletion retinoblastoma (Rb) were submitted to fractionated doses of ? radiations. Irradiation reduced the population doublings in normal fibroblasts and the decline was inversely related to the dose. An increase in population doublings was obtained with one of the Rb cell lines. Foci appeared in the irradiated culture of the other Rb donor. It is suggested that fibroblasts from patients with Rb are able to express some phenotypical properties of transformed cells, perhaps related to factors rendering them more susceptible to carcinogens

13

Extracellular depolymerization of hyaluronic acid in cultured human skin fibroblasts  

International Nuclear Information System (INIS)

The chain length of [3H]hyaluronic acid synthesized by cultivating human skin fibroblasts in the presence of [3H]glucosamine was investigated. [3H]Hyaluronic acid obtained from the matrix fraction was excluded from a Sepharose CL-2B column irrespective of the incubation period, whereas that from the medium was depolymerized into a constant chain length (Mr = 40,000). The reducing and non-reducing terminals of the depolymerized hyaluronic acid were N-acetylglucosamine and glucuronic acid, respectively. Prolonged incubation produced no oligosaccharides as shown by examination of hyaluronidase digests, suggesting the presence of a novel endo-beta-N-acetylglucosaminidase in cultured human skin fibroblasts

14

Studies of the in vivo radiosensitivity of human skin fibroblasts  

International Nuclear Information System (INIS)

Background and purpose: To examine the radiosensitivity of skin cells obtained directly from the irradiated skin of patients undergoing fractionated radiation treatment prior to surgery for treatment of soft tissue sarcoma (STS) and to determine if there was a relationship with the development of wound healing complications associated with the surgery post-radiotherapy. Methods: Micronucleus (MN) formation was measured in cells (primarily dermal fibroblasts) obtained from human skin at their first division after being removed from STS patients during post-radiotherapy surgery (2-9 weeks after the end of the radiotherapy). At the time of radiotherapy (planned tumor dose - 50 Gy in 25 daily fractions) measurements were made of surface skin dose at predetermined marked sites. Skin from these sites was obtained at surgery and cell suspensions were prepared directly for the cytokinesis-blocked MN assay. Cultured strains of the fibroblasts were also established from skin nominally outside the edge of the radiation beam and DNA damage (MN formation) was examined following irradiation in vitro for comparison with the results from the in situ irradiations. Results: Extensive DNA damage (MN) was detectable in fibroblasts from human skin at extended periods after irradiation (2-9 weeks after the end of the 5-week fractionated radiotherapy). Analysis of skin receiving a range of doses demonstrated that the level of damage observed was dose dependent. There was no clear correlatioe dependent. There was no clear correlation between the level of damage observed after irradiation in situ and irradiation of cell strains in culture. Similarly, there was no correlation between the extent of MN formation following in situ irradiation and the propensity for the patient to develop wound healing complications post-surgery. Conclusions: Despite the presence of DNA damage in dermal fibroblasts weeks after the end of the radiation treatment, there was no relationship between this damage and wound healing complications following surgery post-irradiation. These results suggest that factors other than the radiosensitivity of the skin fibroblasts likely also play a role in wound healing in deep wound sites associated with surgery for STS following radiation therapy

15

Oral fibroblasts produce more HGF and KGF than skin fibroblasts in response to co-culture with keratinocytes  

DEFF Research Database (Denmark)

The production of hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF) in subepithelial fibroblasts from buccal mucosa, periodontal ligament, and skin was determined after co-culture with keratinocytes. The purpose was to detect differences between the fibroblast subpopulations that could explain regional variation in epithelial growth and wound healing. Normal human fibroblasts were cultured on polystyrene or maintained in collagen matrix and stimulated with keratinocytes cultured on membranes. The amount of HGF and KGF protein in the culture medium was determined every 24 h for 5 days by ELISA. When cultured on polystyrene, the constitutive level of KGF and HGF in periodontal fibroblasts was higher than the level in buccal and skin fibroblasts. In the presence of keratinocytes, all three types of fibroblasts in general increased their HGF and KGF production 2-3 times. When cells were maintained in collagen, the level of HGF and KGF was decreased mainly in skin cultures. However, in oral fibroblasts, induction after stimulation was at a similar level in collagen compared to on polystyrene. Skin fibroblasts maintained in collagen produced almost no HGF whether with or without stimulation. The results demonstrate that the secretion of KGF and HGF in both unstimulated fibroblasts and in fibroblasts co-cultured with keratinocytes is dependent on the type of fibroblasts. In general, the periodontal fibroblasts had the highest level of cytokine production. This high level of growth factor production may influence the proliferation and the migration of junctional epithelium and thereby influence the development of periodontal disease.

GrØn, Birgitte; Stoltze, Kaj

2002-01-01

16

Proliferation and glycosaminoglycans secretion in fibroblasts from psoriatic skin: differential responses to retinoids.  

Science.gov (United States)

The effects of four retinoids, all-trans-retinoic acid (tretinoin), 13-cis-retinoic acid (isotretinoin), R0 10-1670 (etretin) and the arotinoid, R0 15-0778, on fibroblast proliferation and glycosaminoglycans (GAG) secretion in vitro were studied. Fibroblasts lines cultured from normal skin (HSF) were compared with those from lesional (PSA) and non-lesional (PSB) psoriatic skin. In general, the retinoids inhibited proliferation; the action was cytostatic, in rank order tretinoin greater than isotretinoin greater than etretin greater than arotinoid. The psoriatic cells tended to be more sensitive than the HSF lines, overall mean proliferation values (+/- SEM), as a percentage of untreated controls being: HSF 72 ++- 3, PSA 61 +/- 3 and PSB 54 +/- 3. Stimulation of GAG secretion at low concentrations (10(-7) M) of all four retinoids, declined as concentrations increased, and secretion was inhibited at 10(-4)M in PSB fibroblasts. Calculation of effects on GAG secretion due to changes in cell density confirmed the rank order for direct stimulation of secretion as arotinoid greater than etretin greater than isotretinoin greater than tretinoin. Electrophoresis of [3H]-labelled glycosaminoglycans secreted in the presence of 10(-7) M arotinoid showed that it was predominantly hyaluronic acid, as in untreated cells. These data confirm that different retinoids have contrasting levels of effects on mesenchymal cells and suggest a greater sensitivity to drugs in fibroblasts from psoriatic skin. PMID:2961363

Priestley, G C

1987-11-01

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Proliferation index of camel skin fibroblast cells as nuclear donor  

International Nuclear Information System (INIS)

Jaiselmeri is an excellent breed of riding camel, found in Jaiselmer and other adjoining districts of Western Rajasthan in India. Jaiselmeri camel like other pack animals are declining in India over the years due to increased mechanization and control of desert agriculture to some extent. The deep freezing technology on camel semen is poorly developed in India. The somatic cell technology has been developed at this Institute as an alternative tool of long-term conservation on endangered livestock breeds. For this study, samples of (0.25 cm2) skin tissue were collected from ear biopsy from elite male germplasm from National Research Centre on Camel, Bikaner. Skin tissues were cultured at 37 deg. C in Medium (DMEM+ Ham's F-12 nutritive mixture) supplemented with 10% fetal bovine serum, L-Glutamine and antibiotics in an incubator under 98% humidified and 5% Co2 atmosphere. The cell explants were visible from 12-16 days of culture. The cells were allowed to confluent in the TC flasks for additional 3-5 days till nearly 80% surface area is covered by the cells. The primary cells were harvested by usual trypsin-EDTA protocol. The cells were counted using Neubar's haemocytometer and cells were passaged subsequently. Since no reference values were available for camel skin fibroblasts, the present experiments were conducted to study the cell proliferation index, population doubling time, standard growth curve and cell viability using standard growth and MTT assays. ity using standard growth and MTT assays. It is shown that growth curves showed true sigmoid shape but a marked variation between the cell lines was observed. Moreover, cells, which grew faster attained plateau on day 6 while in slow growing cultures, the curve showed elevation even on day 8. This is probably due to non-availability of growing space for cells having faster growth rate. It was concluded that all animals do not produce karyoplast donors at equal rate or efficiency. Therefore, the growing cultures need to be compared with standard growth curve each time the cells are used as nuclear donor cells for cloning. Cell Proliferation Index: Cell multiplication rates vary considerably under different culture condition and slight change in environment or composition of medium may affect the proliferation of cells significantly. For camel skin fibroblast cells, the standard multiplication rate and the population doubling time was not known earlier. In order to study the proliferative indices of the growing cells using objective parameters, MTT assay was conducted. In this assay, the dividing and viable cells take up MTT [3- (4,5- dimethylthiozol- 2yl) 2,5 diphenyltetrazolium bromide] and a colour is developed. The intensity of colour is measured by ELISA reader at 540-570 nm. For this, 4000 cells per well were seeded in 96 well ELISA plate (flat bottom, Nunc) and cultured at 37 deg. C. First two rows of eight wells each were kept as negative and positive controls respectively. Rest of the 10 rows were kept as treatments. One row was harvested at an interval of 24 hours and adjoining row was treated with MTT solution for 4 hours. The MTT treated cells were fixed in 10% DMSO. Figures 2 and 3 show that the cell proliferation index both in terms of cell count and absorbance values in ELISA reader at appropriate wavelength was similar. From this study it is clear that MTT assay can give fairly accurate figures of cell proliferation rate of skin fibroblasts. Ploidy level: During long-term culture, the cells are likely to develop one or other type of chromosomal abnormalities. It must be ensured that the cells in different passages are checked for normal ploidy so that the viable clones can be developed from them. In order to see the utility of cells from Jaiselmeri camel as nuclear donor, the chromosomal profile was studied following the protocol described elsewhere. The 2N chromosomes up to passage No 4 (15th population doubling) was found to be normal (74XY) in 97% of the cells. From these preliminary studies it appears that camel skin fibroblast cells behave normally i

18

PKC? Deficits in Alzheimer's Disease Brains and Skin Fibroblasts.  

Science.gov (United States)

In Alzheimer's disease (AD) transgenic mice, activation of synaptogenic protein kinase C ? (PKC?) was found to prevent synaptotoxic amyloid-? (A?)-oligomer elevation, PKC? deficits, early synaptic loss, cognitive deficits, and amyloid plaque formation. In humans, to study the role of PKC? in the pathophysiology of AD and to evaluate its possible use as an early AD-biomarker, we examined PKC? and A? in the brains of autopsy-confirmed AD patients (n = 20) and age-matched controls (AC, n = 19), and in skin fibroblast samples from AD (n = 14), non-AD dementia patients (n = 14), and AC (n = 22). Intraneuronal A? levels were measured immunohistochemically (using an A?-specific antibody) in hippocampal pyramidal cells of human autopsy brains. PKC? was significantly lower in the hippocampus and temporal pole areas of AD brains, whereas A? levels were significantly higher. The ratio of PKC? to A? in individual CA1 pyramidal cells was markedly lower in the autopsy AD brains versus controls. PKC? was inversely correlated with A? levels in controls, whereas in AD patients, PKC? showed no significant correlation with A?. In autopsy brains, PKC? decreased as the Braak score increased. Skin fibroblast samples from AD patients also demonstrated a deficit in PKC? compared to controls and an AD-specific change in the A?-oligomer effects on PKC?. Together, these data demonstrate that the relationship between A? levels and PKC? is markedly altered in AD patients' brains and skin fibroblasts, reflecting a loss of protective effect of PKC? against toxic A? accumulation. These changes of PKC? levels in human skin fibroblasts may provide an accurate, non-invasive peripheral AD biomarker. PMID:25125477

Khan, Tapan K; Sen, Abhik; Hongpaisan, Jarin; Lim, Chol S; Nelson, Thomas J; Alkon, Daniel L

2015-01-01

19

Differential degradation of [35S]methionine polypeptides in Duchenne muscular dystrophy skin fibroblasts in vitro.  

OpenAIRE

Rates of protein turnover have been measured in three normal and three Duchenne muscular dystrophy (DMD) skin fibroblast cell lines. Cell populations were analyzed at identical states with regard to cell number, state of topoinhibition, and cumulative population doublings (CPD). Net protein synthesis measured by the incorporation of [35S]methionine in an 18-hr pulse was reduced by an average of 34%; degradation of total cellular protein measured after an 18-hr pulse with [35S]methionine and a...

Rodemann, H. P.; Bayreuther, K.

1986-01-01

20

Skin equivalent tensional force alters keloid fibroblast behavior and phenotype.  

Science.gov (United States)

Skin tension may influence keloid scar behavior, development, and spreading, e.g., butterfly-shaped keloid disease in the sternum. Here, we developed a three-dimensional (3D) in vitro model to mimic in vivo tension and evaluate keloid fibroblast (KF) behavior and extracellular matrix synthesis under tension. In vivo skin tension measured in volunteers (n?=?4) using 3D?image photogrammetry enabled prediction of actual force (35?mN). A novel cell force monitor applied tension in a fibroblast-populated 3D collagen lattice replicating the in vivo force. The effect of tension on keloid (n?=?10) fibroblast (KF) and normal skin (n?=?10) fibroblasts (NF) at set time points (6, 12, and 24 hours) was measured in Hsp27, PAI-2, and ?2?1 integrin, tension-related genes demonstrating significant (p?

Suarez, Edna; Syed, Farhatullah; Rasgado, Teresa A; Walmsley, Alan; Mandal, Parthasarathi; Bayat, Ardeshir

2014-01-01

21

Leopard syndrome associated with hyperelastic skin: analysis of collagen metabolism in cultured skin fibroblasts.  

Science.gov (United States)

We present a patient with Leopard syndrome and hyperelastic skin. Biochemical analysis using cultured skin fibroblasts showed normal type III and V collagen synthesis, lysyl hydroxylation level of type I procollagen and processing of pro-alpha(1) and alpha(2)(I). Our results suggest that molecular defects of hyperelasticity in Leopard syndrome are not related to abnormal collagen metabolism, although not all steps of collagen synthesis have been investigated. PMID:10449938

Ohkura, T; Ohnishi, Y; Kawada, A; Tajima, S; Ishibashi, A; Ono, K

1999-01-01

22

Abnormal collagen metabolism in cultured skin fibroblasts from patients with Duchenne muscular dystrophy.  

OpenAIRE

Total collagen synthesis is decreased by about 29% (P less than 0.01) in skin fibroblasts established in vitro from male patients with Duchenne muscular dystrophy (DMD) as compared with that in normal male skin fibroblasts in vitro. The reduction in collagen synthesis is associated with an approximately 2-fold increase in collagen degradation in DMD fibroblasts. Correlated to these alterations in the metabolism of collagen, DMD fibroblasts express a significantly higher hydroxyproline/proline...

Rodemann, H. P.; Bayreuther, K.

1984-01-01

23

Human skin fibroblasts in vitro differentiate along a terminal cell lineage.  

OpenAIRE

Secondary mitotic human skin fibroblast populations in vitro underwent 53 +/- 6 cumulative population doublings (CPD) in 302 +/- 27 days. When the growth capacity of the mitotic fibroblasts is exhausted, and if appropriate methods are applied, the fibroblasts differentiate spontaneously into postmitotic fibroblast populations, which were kept in stationary culture for up to 305 +/- 41 additional days. Mitotic and postmitotic fibroblast populations are heterogeneous populations with reproducib...

Bayreuther, K.; Rodemann, H. P.; Hommel, R.; Dittmann, K.; Albiez, M.; Francz, P. I.

1988-01-01

24

Aluminum is More Cytotoxic than Lunar Dust in Human Skin and Lung Fibroblasts  

Science.gov (United States)

NASA plans to build a permanent space station on the moon to explore its surface. The surface of the moon is covered in lunar dust, which consists of fine particles that contain silicon, aluminum and titanium, among others. Because this will be a manned base, the potential toxicity of this dust has to be studied. Also, toxicity standards for potential exposure have to be set. To properly address the potential toxicity of lunar dust we need to understand the toxicity of its individual components, as well as their combined effects. In order to study this we compared NASA simulant JSC-1AVF (volcanic ash particles), that simulates the dust found on the moon, to aluminum, the 3rd most abundant component in lunar dust. We tested the cytotoxicity of both compounds on human lung and skin fibroblasts (WTHBF-6 and BJhTERT cell lines, respectively). Aluminum oxide was more cytotoxic than lunar dust to both cell lines. In human lung fibroblasts 5, 10 and 50 g/sq cm of aluminum oxide induced 85%, 61% and 30% relative survival, respectively. For human skin fibroblasts the same concentrations induced 58%, 41% and 58% relative survival. Lunar dust was also cytotoxic to both cell lines, but its effects were seen at higher concentrations: 50, 100, 200 and 400 g/sq cm of lunar dust induced a 69%, 46%, 35% and 30% relative survival in the skin cells and 53%, 16%, 8% and 2% on the lung cells. Overall, for both compounds, lung cells were more sensitive than skin cells. This work was supported by a NASA EPSCoR grant through the Maine Space Grant Consortium (JPW), the Maine Center for Toxicology and Environmental Health., a Fulbright Grant (JM) and a Delta Kappa Gamma Society International World Fellowship (JM).

Hammond, D.; Shehata, T.; Hammond, D.; Shehata, T.; Wise, J.P.; Martino, J; Wise, J.P.; Wise, J.P.

2009-01-01

25

Lysophosphatidic acid-activated Cl? current activity in human systemic sclerosis skin fibroblasts  

OpenAIRE

Objectives. SSc (scleroderma) is an often fatal disease characterized by widespread tissue fibrosis. Fibroblasts play a key role in SSc-associated fibrosis. This study was designed to determine: (i) whether fibroblasts isolated from skin of patients with SSc have increased lysophosphatidic acid-activated Cl? current (IClLPA) activity vs healthy controls; (ii) whether myofibroblast differentiation is involved in SSc skin fibrosis; and (iii) whether SSc fibroblasts have different proliferatio...

Yin, Zhaohong; Carbone, Laura D.; Gotoh, Mari; Postlethwaite, Arnold; Bolen, Alyssa L.; Tigyi, Gabor J.; Murakami-murofushi, Kimiko; Watsky, Mitchell A.

2010-01-01

26

Effect of resveratrol on cultured skin fibroblasts from patients with oxidative phosphorylation defects.  

Science.gov (United States)

Few therapeutic options are available to patients with oxidative phosphorylation disorders. Administering pharmacological agents that are able to stimulate mitochondrial biogenesis have been put forward as a possible treatment, yet the approach remains in need of thorough testing. We investigated the effect of resveratrol in an in vitro setting. Mitochondrial enzymatic activities were tested in cultured skin fibroblasts from patients harboring a nuclear defect in either complex II or complex IV (n?=?11), and in fibroblasts from healthy controls (n?=?11). In the latter, preincubation with resveratrol resulted in a significant increase of citrate synthase, complex II and complex IV enzyme activity. In patients with complex II or complex IV deficiency, however, activity of the deficient complex could not be substantially augmented, and response was dependent upon the residual activity. We conclude that resveratrol is not capable of normalizing oxidative phosphorylation activities in deficient cell lines. PMID:23620374

De Paepe, Boel; Vandemeulebroecke, Katrien; Smet, Joél; Vanlander, Arnaud; Seneca, Sara; Lissens, Willy; Van Hove, Johan Lk; Deschepper, Ellen; Briones, Paz; Van Coster, Rudy

2014-02-01

27

Lysosomal Storage Causes Cellular Dysfunction in Mucolipidosis II Skin Fibroblasts*  

Science.gov (United States)

Mucolipidosis II (ML-II) is a fatal inherited metabolic disease caused by deficiency of GlcNAc-phosphotransferase, which plays a role in generating the mannose 6-phosphate recognition marker on lysosomal enzymes. In ML-II, many lysosomal acid hydrolases are mistargeted out of cells, and lysosomes become filled with undigested substrates, which explains inclusion cell disease as an alternative name for this disease. In this study, we revealed various cellular phenotypes in ML-II skin fibroblasts. We quantitated phospholipid and cholesterol within cells and showed ?2-fold accumulation in ML-II as compared with normal cells. Lysosomal pH of ML-II cells was higher than that of normal cells (5.29 ± 0.08 versus 4.79 ± 0.10, p < 0.001). The proliferated lysosomes in ML-II cells were accumulated ?3-fold in amount as compared with normal cells. Intracellular logistics including endocytosis and mannose 6-phosphate receptor recycling were impaired in ML-II cells. To confirm whether these ML-II cellular phenotypes derive from deficient lysosomal acid hydrolases within lysosomes, we performed supplementation of lysosomal enzymes using a partially purified total enzyme mixture, which was derived from the conditioned culture medium of normal skin fibroblasts after NH4Cl treatment. This supplementation corrected all of the previously described ML-II phenotypes. In addition, the autophagic and mitochondrial impairment that we have previously reported improved, and inclusion bodies disappeared on electron micrography following total lysosomal enzyme supplementation. Our results indicate that various cellular phenotypes in ML-II are caused by the deficiency of many lysosomal enzymes and massive accumulation of undigested substrates. PMID:21846724

Otomo, Takanobu; Higaki, Katsumi; Nanba, Eiji; Ozono, Keiichi; Sakai, Norio

2011-01-01

28

Normal skin and hypertrophic scar fibroblasts differentially regulate collagen and fibronectin expression as well as mitochondrial membrane potential in response to basic fibroblast growth factor  

OpenAIRE

Basic fibroblast growth factor (bFGF) regulates skin wound healing; however, the underlying mechanism remains to be defined. In the present study, we determined the effects of bFGF on the regulation of cell growth as well as collagen and fibronectin expression in fibroblasts from normal human skin and from hypertrophic scars. We then explored the involvement of mitochondria in mediating bFGF-inducedeffects on the fibroblasts. We isolated and cultivated normal and hypertrophic scar fibroblasts...

Rui Song; Hui-Ning Bian; Wen Lai; Hua-De Chen; Ke-Seng Zhao

2011-01-01

29

Diversity of fibroblasts--a review on implications for skin tissue engineering.  

Science.gov (United States)

Enormous advances in the development of skin substitutes have occurred in the past 3 decades. Major obstacles yet to be overcome in the quest for an optimal skin substitute include controlling scar formation, contraction and the loss of adnexal structures. Mesenchyme-derived signals are essential for epithelial proliferation, skin morphogenesis, homeostasis and differentiation. Having previously shown that fibroblasts differentiate along a lineage from highly proliferative progenitor fibroblasts with characteristic spindle-shaped appearance to differentiated postmitotic polygonal fibrocytes, we have now established that the different subsets of fibroblasts exert significantly different patterns of cytokine release and that the highest levels of keratinocyte growth factor and transforming growth factor-beta1 expression result from differentiated fibroblasts. Coculture studies with keratinocytes reveal that postmitotic fibroblasts stimulate keratinocyte proliferation to a greater extent than progenitor fibroblasts. Acellular and fibroblast-seeded dermal substitutes have been shown to improve scarring and contraction in animal studies, the latter substitutes yielding the most favorable results. Fibroblasts from different body sites display different functional properties which may affect their suitability for dermal substitutes. Future in vivo human studies in tissue-engineered dermal substitutes will likely focus on fibroblast-seeded lattices and the impact of fibroblast subpopulations and bone marrow-derived mesenchymal stem cells on dermal regeneration. PMID:18042973

Nolte, Sonja Veronika; Xu, Weiguo; Rennekampff, Hans-Oliver; Rodemann, H Peter

2008-01-01

30

Abnormal Collagen Metabolism in Cultured Skin Fibroblasts from Patients with Duchenne Muscular Dystrophy  

Science.gov (United States)

Total collagen synthesis is decreased by about 29% (P skin fibroblasts established in vitro from male patients with Duchenne muscular dystrophy (DMD) as compared with that in normal male skin fibroblasts in vitro. The reduction in collagen synthesis is associated with an approximately 2-fold increase in collagen degradation in DMD fibroblasts. Correlated to these alterations in the metabolism of collagen, DMD fibroblasts express a significantly higher hydroxyproline/proline ratio (DMD: 1.36-1.45; P < 0.01) than do normal fibroblasts (controls: 0.86-0.89). The increased hydroxylation of proline residues of collagen (composed of type I and type III) could be the cause for the enhanced degradation of collagen in DMD fibroblasts.

Rodemann, H. Peter; Bayreuther, Klaus

1984-08-01

31

Radiation effects on the concentration of SOD and cathepsin D activities in cultured skin fibroblasts  

International Nuclear Information System (INIS)

Human skin fibroblasts were irradiated with 60Co ?-ray 10 Gy?40 Gy. After one hour, the concentration of the oxygen free radicals increased significantly, the activity of SOD in cultured fibroblasts decreased markedly, and the activity of cathepsin D increased significantly. These changes related with the dose. when the radiation dose within the range of 0?40 Gy, the the concentration of the oxygen free radicals related with the activity of SOD and cathepsin D in cultured sin fibroblasts

32

Mutagenic effects of alpha particles in normal human skin fibroblasts  

International Nuclear Information System (INIS)

Alpha-irradiation to the bronchial airways from inhaled radon progeny increases the risk of developing lung cancer. The molecular mechanism of radon-induced lung cancer is not clear, but one of the most important genetic effects of ionizing radiation is the induction of gene mutation. Mutations, especially those associated with visible chromosome abnormalities in humans, have been associated with cancer. Therefore, our objective is to use a well-defined model system to determine the mutagenic potential of alpha particles in normal human skin cells and to define this action at the molecular level. Normal human skin fibroblasts were irradiated with alpha particles (3.59 MeV, LET 115 keV ?m-1) emitted from the decay of 238Pu. Mutagenicity was determined at the X-linked hypoxanthine guanine phosphoribosyl transferase (HPRT) locus. Results from this study indicate that beta particles were more efficient in mutation induction than gamma rays. Based on the initial slopes of the dose-response curves, the RBE for mutation is about 8 for alpha particles. HPRT-deficient mutants which are resistant to 6-thioguanine have been isolated and analyzed by the Southern blot technique. To date, we have characterized 69 gamma-ray-induced and 195 alpha-particle-induced HPRT-deficient mutants. Our data indicate that more than 50% of all gamma-ray-induced mutants have band patterns identical to that observed for the normal structural HPRT gene, whereas the remaining mutural HPRT gene, whereas the remaining mutants (45%) contain either a rearrangement, partial deletion, or total deletion of the HPRT gene. In contrast, only 30% of alpha-particle-induced human HPRT mutants contain a normal Southern blot pattern, and about 50% indicate total deletion of the HPRT gene. Our results support the notion that high-LET radiation produces more unrepaired or misrepaired DNA damage than do gamma rays

33

Colony size distributions according to in vitro aging in human skin fibroblasts  

International Nuclear Information System (INIS)

To investigate the percentage of colonies with 16 or more cells distribution of human skin fibroblast according to in vitro aging, and to evaluate the relationship between percentage of colonies with 16 or more cells and in vivo donor age in human skin fibroblast culture. C1, C2, C3a, and C3b human skin fibroblast samples from three breast cancer patients were used as subjects. The C1, C2, and C3a donor were 44, 54, and 55 years old, respectively. C3a and C3b cells were isolated from the same person. Single cell suspension of skin fibroblasts was prepared with primary explant technique. One hundred cells are plated into 100ml tissue culture flask and cultured for two weeks. The colony size was defined as colonies with 16 or more cells. The cultured cell was stained with crystal violet, and number of cells in each colony was determined with stereo microscope at x 10 magnification. Passage number of C1, C2, C3a and C3b skin fibroblast were 12th, 17th, and 14th, respectively. Percentage of colonies with 16 or more cells of skin fibroblast samples decreased with increasing in vitro passage number. In contrast, cumulative population doublings of skin fibroblast sample increased with increasing in vitro passage number. Percentage of colonies with 16 or more cells also decreased with increasing population doublings in human skin fibroblast culture. There was strong correlation with percentage of colonised with 16 or more cells and population doublings in C3a skin fibroblast pulation doublings in C3a skin fibroblast sample. At the same point of population doublings, the percentage of colonies with 16 or more cells of the young C1 donor was higher level than the old C3a donor. The population doublings increased with increasing in vitro passage number but percentage of colonies with 16 or more cells decreased. The results of this study imply that percentage of colonies with 16 or more cells is useful as a indicator of in vitro human skin fibroblast aging and may estimate the in vivo donor age

34

Coxsackievirus B3 infection alters plasma membrane of neonatal skin fibroblasts.  

OpenAIRE

Replication of coxsackievirus B3 occurred for days in cultures of murine neonatal skin fibroblasts in the absence of cytopathology and resulted in alteration of the plasma membrane. Dual immunofluorescence studies showed that the lectin Ulex europaeus agglutinin I bound only to cells producing viral capsid antigens. Cultures of coxsackievirus B3-inoculated murine neonatal skin fibroblasts showed maximum binding of this lectin at 72 h postinoculation. These data show that in a nonlytic infecti...

Lutton, C. W.; Gauntt, C. J.

1986-01-01

35

Establishment and biological characteristics comparison of Chinese swamp buffalo (Bubalus bubalis) fibroblast cell lines.  

Science.gov (United States)

To establish fibroblast cell lines from different tissues and to compare the biological characteristics of those cell lines, five fibroblast cell lines derived from Chinese swamp buffalo (Bubalus bubalis) were selected for comparative assays. Cell style and survival rate (before cryogenic preservation and after recovery) were tested, and karyotype, patterns of isoenzymes of lactic dehydrogenase, malic dehydrogenase, and cell cycle were analyzed. These cell lines had a healthy morphology with a typical spindle shape, and assessment of cell style showed these cells to be very pure fibroblasts. Cell growth curves showed a typical "S" shape. Results of microorganism contamination assays were negative, and isoenzyme analysis showed no cross-contamination. The number of chromosomes (2n) of swamp buffalo is 48. Between 28% and 46% of the cells were 2n, and cell apoptosis was not pronounced at 20th generation. Results showed that skin fibroblasts were more adaptable to tissue culture conditions than the ones from kidneys and ear margin, and they are more suitable for cellular manipulation in Chinese swamp buffalo. PMID:23990385

Luo, Jun; Liang, Ming-ming; Yang, Xiao-gan; Xu, Hui-yan; Shi, De-shun; Lu, Sheng-sheng

2014-01-01

36

Senescent phenotypes of skin fibroblasts from patients with Tangier disease  

International Nuclear Information System (INIS)

Tangier disease (TD) is characterized by a deficiency of high density lipoprotein (HDL) in plasma and patients with TD have an increased risk for coronary artery disease (CAD). Recently, we reported that fibroblasts from TD exhibited large and flattened morphology, which is often observed in senescent cells. On the other hand, data have accumulated to show the relationship between cellular senescence and development of atherosclerotic CAD. The aim of the present study was to investigate whether TD fibroblasts exhibited cellular senescence. The proliferation of TD fibroblasts was gradually decreased at population doubling level (PDL) ?10 compared with control cells. TD cells practically ceased proliferation at PDL ?30. DNA synthesis was markedly decreased in TD fibroblasts. TD cells exhibited a higher positive rate for senescence-associated ?-galactosidase (SA-?-gal), which is one of the biomarkers of cellular senescence in vitro. These data showed that TD cells reached cellular senescence at an earlier PDL compared with controls. Although, there was no difference in the telomere length of fibroblasts between TD and controls at the earlier passage (PDL 6), the telomere length of TD cells was shorter than that of controls at the late passage (PDL 25). Taken together, the current study demonstrates that the late-passaged TD fibroblasts showed senescent phenotype in vitro, which might be related to the increased cardiovascular manifestations in TD patientsifestations in TD patients

37

Hyaluronan uptake by adult human skin fibroblasts in vitro  

Directory of Open Access Journals (Sweden)

Full Text Available Low and high molecular weight hyaluronan (HA was added to adult human fibroblasts grown in monolayer to assess its influence on CD44 expression, its internalisation and effect on cell growth. CD44 expression on the surface of in vitro fibroblasts was not modified by different concentrations of FCS, whereas it was sensitive to cell cycle, being higher in the growing than in the resting phase. Independently from molecular weight, upon addition of exogenous HA (from 0.1 up to 1 mg/mL to fibroblasts in the growing phase, a slight but constant decrease of the expression of CD44 on the surface of fibroblasts was observed; moreover, HA induced a rearrangement of CD44 into patches in close relationship with the terminal regions of stress fibers, which became thicker and more rigid after a few hours from the addition of HA to the medium. Fluorescent HA, added to the culture medium, rapidly attached to the plasma membrane and in less than two minutes was observed within cells, partly in association with its receptor CD44. By the contemporary use of neutral red, which accumulates into functional lysosomes, the great majority of internalised HA was found within lysosomes. HA receptor RHAMM-IHABP was rather homogeneously localised within the cytoplasm of normal growing fibroblasts. Upon addition of HA, the RHAMM-IHABP distribution became discontinuous around the nucleus. Addition of HA to fibroblasts induced a significant inhibition of cell growth, which was dependent on HA concentration and irrespective of HA molecular weight, at least in the ranges tested. Results show that extra-cellular HA is rapidly taken up by human dermal fibroblasts together with its CD44 receptor, and transported mostly to the lysosomes. Both low and high molecular weight HA induced down-regulation of cell proliferation, which would seem to be mediated by HA catabolism.

MA Croce

2009-06-01

38

Trisomy 13 mosaicism demonstrated only in skin fibroblasts in a patient presenting psychomotor retardation, pigmentary dysplasia and some dysmorphic features  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english A Brazilian female infant presented delayed psychomotor development, skin pigmentary dysplasia and some dysmorphic features. Chromosome analysis from peripheral blood culture was normal, but the karyotype from skin fibroblasts revealed mosaicism for trisomy 13. This case demonstrates the relevance o [...] f performing chromosomal analysis of skin fibroblasts in patients with mental retardation, associated with pigmentary dysplasia of the skin and a normal karyotype in peripheral blood lymphocytes. To our knowledge, it is the first report of trisomy 13 demonstrated only in skin fibroblasts.

A.P.S., Ferreira; L.F., Mazzucatto; E.S., Ramos; J.M., Pina-Neto.

39

Trisomy 13 mosaicism demonstrated only in skin fibroblasts in a patient presenting psychomotor retardation, pigmentary dysplasia and some dysmorphic features  

Directory of Open Access Journals (Sweden)

Full Text Available A Brazilian female infant presented delayed psychomotor development, skin pigmentary dysplasia and some dysmorphic features. Chromosome analysis from peripheral blood culture was normal, but the karyotype from skin fibroblasts revealed mosaicism for trisomy 13. This case demonstrates the relevance of performing chromosomal analysis of skin fibroblasts in patients with mental retardation, associated with pigmentary dysplasia of the skin and a normal karyotype in peripheral blood lymphocytes. To our knowledge, it is the first report of trisomy 13 demonstrated only in skin fibroblasts.

A.P.S. Ferreira

1996-01-01

40

Enhanced reactivation of ultraviolet-damaged herpes virus in ultraviolet pretreated skin fibroblasts of cancer prone donors  

International Nuclear Information System (INIS)

An enhanced reactivation of ultraviolet-damaged (u.v. at 254 nm) unclear replicating double-stranded DNA viruses occurs when corresponding host cells are treated with radiation or carcinogens prior to infection. This phenomenon seems to be due to an induced DNA repair activity the nature of which is yet unknown. The u.v.-induced enhanced reactivation (ER) of u.v.-damaged herpes simplex virus (u.v. - HSV) was compared in dividing skin fibroblasts of 30 donors either normal or afflicted by genetic disorders, some of which confer a high risk for sunlight induced skin cancers. Cultures were exposed to a single dose of 1.0-25 J.m-2 from 0-60 h before infection with u.v.-HSV (at about 10-3 survival) and the rate of viral production was determined. ER was maximal for a 36 h time interval in all lines. The u.v. dose eliciting maximal ER was 15 J.m-2 in fibroblasts from normal donors, xeroderma pigmentosum (XP) heterozygotes, Mibelli's porokeratosis, diffused naevomatosis, Down's syndrome, xerodermoids, XP variants and epidermodysplasia verruciformis. However, in the latter 3 cases, ER was almost 10 times more pronounced than in the normal cases. The u.v. dose eliciting maximal ER was 0.1, 0.3 and 2 J.m-2 in excision deficient XP fibroblasts from groups A, D and C, respectively, 2.5 J.m-2 in 11961 fibroblasts and 5 J.m-2 in fibroblast lines from cockayne s syndrome

41

Distinct fibroblast lineages determine dermal architecture in skin development and repair.  

Science.gov (United States)

Fibroblasts are the major mesenchymal cell type in connective tissue and deposit the collagen and elastic fibres of the extracellular matrix (ECM). Even within a single tissue, fibroblasts exhibit considerable functional diversity, but it is not known whether this reflects the existence of a differentiation hierarchy or is a response to different environmental factors. Here we show, using transplantation assays and lineage tracing in mice, that the fibroblasts of skin connective tissue arise from two distinct lineages. One forms the upper dermis, including the dermal papilla that regulates hair growth and the arrector pili muscle, which controls piloerection. The other forms the lower dermis, including the reticular fibroblasts that synthesize the bulk of the fibrillar ECM, and the preadipocytes and adipocytes of the hypodermis. The upper lineage is required for hair follicle formation. In wounded adult skin, the initial wave of dermal repair is mediated by the lower lineage and upper dermal fibroblasts are recruited only during re-epithelialization. Epidermal ?-catenin activation stimulates the expansion of the upper dermal lineage, rendering wounds permissive for hair follicle formation. Our findings explain why wounding is linked to formation of ECM-rich scar tissue that lacks hair follicles. They also form a platform for discovering fibroblast lineages in other tissues and for examining fibroblast changes in ageing and disease. PMID:24336287

Driskell, Ryan R; Lichtenberger, Beate M; Hoste, Esther; Kretzschmar, Kai; Simons, Ben D; Charalambous, Marika; Ferron, Sacri R; Herault, Yann; Pavlovic, Guillaume; Ferguson-Smith, Anne C; Watt, Fiona M

2013-12-12

42

Cell surface abnormality in clones of skin fibroblasts from a carrier of Duchenne muscular dystrophy.  

OpenAIRE

We have previously reported that skin fibroblasts from patients with Duchenne muscular dystrophy (DMD) have a lower intercellular adhesiveness than control cells, and that cells from carriers of DMD have normal adhesiveness instead of the expected intermediate value. We have now cloned skin fibroblasts from a carrier of DMD (subject AS) who is also heterozygous for G6PD B/G6PD Mediterranean and determined the intercellular adhesiveness and G6PD phenotypes of the clones. G6PD activity was dete...

Hillier, J.; Jones, G. E.; Statham, H. E.; Witkowski, J. A.; Dubowitz, V.

1985-01-01

43

The effect of keratinocytes on the biomechanical characteristics and pore microstructure of tissue engineered skin using deep dermal fibroblasts.  

Science.gov (United States)

Fibrosis affects most organs, it results in replacement of normal parenchymal tissue with collagen-rich extracellular matrix, which compromises tissue architecture and ultimately causes loss of function of the affected organ. Biochemical pathways that contribute to fibrosis have been extensively studied, but the role of biomechanical signaling in fibrosis is not clearly understood. In this study, we assessed the effect keratinocytes have on the biomechanical characteristics and pore microstructure of tissue engineered skin made with superficial or deep dermal fibroblasts in order to determine any biomaterial-mediated anti-fibrotic influences on tissue engineered skin. Tissue engineered skin with deep dermal fibroblasts and keratinocytes were found to be less stiff and contracted and had reduced number of myofibroblasts and lower expression of matrix crosslinking factors compared to matrices with deep fibroblasts alone. However, there were no such differences between tissue engineered skin with superficial fibroblasts and keratinocytes and matrices with superficial fibroblasts alone. Also, tissue engineered skin with deep fibroblasts and keratinocytes had smaller pores compared to those with superficial fibroblasts and keratinocytes; pore size of tissue engineered skin with deep fibroblasts and keratinocytes were not different from those matrices with deep fibroblasts alone. A better understanding of biomechanical characteristics and pore microstructure of tissue engineered skin may prove beneficial in promoting normal wound healing over pathologic healing. PMID:25176070

Varkey, Mathew; Ding, Jie; Tredget, Edward E

2014-12-01

44

Normal skin and hypertrophic scar fibroblasts differentially regulate collagen and fibronectin expression as well as mitochondrial membrane potential in response to basic fibroblast growth factor  

Directory of Open Access Journals (Sweden)

Full Text Available Basic fibroblast growth factor (bFGF regulates skin wound healing; however, the underlying mechanism remains to be defined. In the present study, we determined the effects of bFGF on the regulation of cell growth as well as collagen and fibronectin expression in fibroblasts from normal human skin and from hypertrophic scars. We then explored the involvement of mitochondria in mediating bFGF-inducedeffects on the fibroblasts. We isolated and cultivated normal and hypertrophic scar fibroblasts from tissue biopsies of patients who underwent plastic surgery for repairing hypertrophic scars. The fibroblasts were then treated with different concentrations of bFGF (ranging from 0.1 to 1000 ng/mL. The growth of hypertrophic scar fibroblasts became slower with selective inhibition of type I collagen production after exposure to bFGF. However, type III collagen expression was affected in both normal and hypertrophic scar fibroblasts. Moreover, fibronectin expression in the normal fibroblasts was up-regulated after bFGF treatment. bFGF (1000 ng/mL also induced mitochondrial depolarization in hypertrophic scar fibroblasts (P < 0.01. The cellular ATP level decreased in hypertrophic scar fibroblasts (P < 0.05, while it increased in the normal fibroblasts following treatment with bFGF (P < 0.01. These data suggest that bFGF has differential effects and mechanisms on fibroblasts of the normal skin and hypertrophic scars, indicating that bFGF may play a role in the early phase of skin wound healing and post-burn scar formation.

Rui Song

2011-05-01

45

Transformation of Human Mesenchymal Cells and Skin Fibroblasts into Hematopoietic Cells  

OpenAIRE

Patients with prolonged myelosuppression require frequent platelet and occasional granulocyte transfusions. Multi-donor transfusions induce alloimmunization, thereby increasing morbidity and mortality. Therefore, an autologous or HLA-matched allogeneic source of platelets and granulocytes is needed. To determine whether nonhematopoietic cells can be reprogrammed into hematopoietic cells, human mesenchymal stromal cells (MSCs) and skin fibroblasts were incubated with the demethylating agent 5-...

Harris, David M.; Hazan-haley, Inbal; Coombes, Kevin; Bueso-ramos, Carlos; Liu, Jie; Liu, Zhiming; Li, Ping; Ravoori, Murali; Abruzzo, Lynne; Han, Lin; Singh, Sheela; Sun, Michael; Kundra, Vikas; Kurzrock, Razelle; Estrov, Zeev

2011-01-01

46

Hyaluronan synthase 2 protects skin fibroblasts against apoptosis induced by environmental stress.  

Science.gov (United States)

A balanced turnover of dermal fibroblasts is crucial for structural integrity and normal function of the skin. During recovery from environmental injury (such as UV exposure and physical wounding), apoptosis is an important mechanism regulating fibroblast turnover. We are interested in the role that hyaluronan (HA), an extracellular matrix molecule synthesized by HA synthase enzymes (Has), plays in regulating apoptosis in fibroblasts. We previously reported that Has1 and Has3 double knock-out (Has1/3 null) mice show accelerated wound closure and increased numbers of fibroblasts in the dermis. In the present study, we report that HA levels and Has2 mRNA expression are higher in cultured Has1/3 null primary skin fibroblasts than in wild type (WT) cells. Apoptosis induced by two different environmental stressors, UV exposure and serum starvation (SS), was reduced in the Has1/3 null cells. Hyaluronidase, added to cultures to remove extracellular HA, surprisingly had no effect upon apoptotic susceptibility to UVB or SS. However, cells treated with 4-methylumbelliferone to inhibit HA synthesis were sensitized to apoptosis induced by SS or UVB. When fibroblasts were transfected with Has2-specific siRNA that lowered Has2 mRNA and HA levels by 90%, both Has1/3 null and WT cells became significantly more sensitive to apoptosis. The exogenous addition of high molecular weight HA failed to reverse this effect. We conclude that Has1/3 null skin fibroblasts (which have higher levels of Has2 gene expression) are resistant to stress-induced apoptosis. PMID:25266724

Wang, Yan; Lauer, Mark E; Anand, Sanjay; Mack, Judith A; Maytin, Edward V

2014-11-14

47

Zinc and Propolis Reduces Cytotoxicity and Proliferation in Skin Fibroblast Cell Culture: Total Polyphenol Content and Antioxidant Capacity of Propolis  

OpenAIRE

It has been demonstrated that zinc exerts its beneficial influence on skin fibroblasts. Propolis, a complex mixture of plant-derived and bees’ products, was reported to stimulate cicatrization processes in skin and prevent infections. The aim of this study was to find out how zinc and propolis influence human skin fibroblasts in cell culture and to compare the effect of individual compounds to the effect of a mixture of zinc and propolis. In this study, zinc, as zinc aspartate, at a concent...

Tyszka-czochara, Ma?gorzata; Pas?ko, Pawe?; Reczyn?ski, Witold; Szlo?sarczyk, Marek; Bystrowska, Beata; Opoka, W?odzimierz

2014-01-01

48

Skin-derived fibroblasts from long-lived species are resistant to some, but not all, lethal stresses and to the mitochondrial inhibitor rotenone  

OpenAIRE

Fibroblast cell lines were developed from skin biopsies of eight species of wild-trapped rodents, one species of bat, and a group of genetically heterogeneous laboratory mice. Each cell line was tested in vitro for their resistance to six varieties of lethal stress, as well as for resistance to the nonlethal metabolic effects of the mitochondrial inhibitor rotenone and of culture at very low glucose levels. Standard linear regression of species-specific lifespan against each species mean stre...

Harper, James M.; Salmon, Adam B.; Leiser, Scott F.; Galecki, Andrzej T.; Miller, Richard A.

2006-01-01

49

Multi-layered culture of human skin fibroblasts and keratinocytes through three-dimensional freeform fabrication.  

Science.gov (United States)

We present a method to create multi-layered engineered tissue composites consisting of human skin fibroblasts and keratinocytes which mimic skin layers. Three-dimensional (3D) freeform fabrication (FF) technique, based on direct cell dispensing, was implemented using a robotic platform that prints collagen hydrogel precursor, fibroblasts and keratinocytes. A printed layer of cell-containing collagen was crosslinked by coating the layer with nebulized aqueous sodium bicarbonate. The process was repeated in layer-by-layer fashion on a planar tissue culture dish, resulting in two distinct cell layers of inner fibroblasts and outer keratinocytes. In order to demonstrate the ability to print and culture multi-layered cell-hydrogel composites on a non-planar surface for potential applications including skin wound repair, the technique was tested on a poly(dimethylsiloxane) (PDMS) mold with 3D surface contours as a target substrate. Highly viable proliferation of each cell layer was observed on both planar and non-planar surfaces. Our results suggest that organotypic skin tissue culture is feasible using on-demand cell printing technique with future potential application in creating skin grafts tailored for wound shape or artificial tissue assay for disease modeling and drug testing. PMID:19108884

Lee, Wonhye; Debasitis, Jason Cushing; Lee, Vivian Kim; Lee, Jong-Hwan; Fischer, Krisztina; Edminster, Karl; Park, Je-Kyun; Yoo, Seung-Schik

2009-03-01

50

Normal skin and hypertrophic scar fibroblasts differentially regulate collagen and fibronectin expression as well as mitochondrial membrane potential in response to basic fibroblast growth factor  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Basic fibroblast growth factor (bFGF) regulates skin wound healing; however, the underlying mechanism remains to be defined. In the present study, we determined the effects of bFGF on the regulation of cell growth as well as collagen and fibronectin expression in fibroblasts from normal human skin a [...] nd from hypertrophic scars. We then explored the involvement of mitochondria in mediating bFGF-inducedeffects on the fibroblasts. We isolated and cultivated normal and hypertrophic scar fibroblasts from tissue biopsies of patients who underwent plastic surgery for repairing hypertrophic scars. The fibroblasts were then treated with different concentrations of bFGF (ranging from 0.1 to 1000 ng/mL). The growth of hypertrophic scar fibroblasts became slower with selective inhibition of type I collagen production after exposure to bFGF. However, type III collagen expression was affected in both normal and hypertrophic scar fibroblasts. Moreover, fibronectin expression in the normal fibroblasts was up-regulated after bFGF treatment. bFGF (1000 ng/mL) also induced mitochondrial depolarization in hypertrophic scar fibroblasts (P

Rui, Song; Hui-Ning, Bian; Wen, Lai; Hua-De, Chen; Ke-Seng, Zhao.

2011-05-01

51

209 cloned embryonic development and gene expression of spontaneously immortalized porcine skin fibroblasts.  

Science.gov (United States)

Immortalization of somatic cells by oncogenes is effective for evaluation of gene targeting tools or molecular cell pathway because of relatively longer life span than normal cells. The BMI1 gene has been used to immortalize human and murine somatic cells, and we confirmed that BMI1 increased cell-life span of porcine skin fibroblasts and had no detrimental effect on pre-implantational development after somatic-cell nuclear transfer (SCNT; unpublished data). Here, we report a primary cell-line which was spontaneously immortalized without transduction of any additional gene. The minipig skin fibroblasts (passage 3 after primary culture) were divided into two parts, one group was electroporetically transfected with pCAG-BMI1-T2A-RFP plasmids (BC1) and the other had no treatment (CC1). To establish the single-cell-originated cell-lines, cells of each group were plated in 100-mm dishes at 100 cells/dish and well-formed colonies were picked up after 2 weeks. These colonies were expanded until they were fully confluent in 100-mm dishes (designated Passage 0) and then, the cell were maintained with DMEM (20% FBS) at 3×10(5) cells/60mm dish. Population doubling time was checked every 5 passages until Passage 45 (sub-cultured during ~160 days) by calculation of cells that had been plated in 12-well plates at 4×10(4) cells/well. The expressions of p16, p21, and DNMT3b genes were determined by RT-qPCR at early (Passage 5) and late (Passage 45) stage. Also, SCNT (Song et al. 2009 Mol. Reprod. Dev. 76, 611-619) embryos using cells of early and middle (Passage 35) stage were evaluated in terms of reprograming efficiency. Data were analysed by t-test (Prism version 5.01, GraphPad Software, La Jolla, CA, USA). While the mean doubling time of BC1 was 25.3h until Passage 40 and drastically increased at Passage 45 (81.7h), that of CC1 was maintained until Passage 45 (mean 38.3h). Expression of p16 in CC1 was significantly higher than that in BC1 at all stages. However, in late stage CC1, p21 expression was significantly lower than other groups and DNMT3b expression was increased. In SCNT embryos, the rate of blastocysts with early stage CC1 (18.3%) was not different from that of early stage BC1 (19.9%). And, although the rates of SCNT blastocyst derived from middle stage cells were decreased than those of early stage cells, there was no difference between BC1 and CC1 (6.6% and 5.4%, respectively). In karyotyping, while BC1 was trisomy of chromosome 14 only in late stage, CC1 had an isochromosome in chromosome 17 from early stage and an additional part was attached in chromosome 11 at late stage CC1. In summary, spontaneously immortalized skin fibroblasts could maintain the cell-life span by down-regulating the p21 expression and the pre-implantational development after SCNT was not different from that of BMI1-immortalized cells. And, additional studies are needed to confirm whether the chromosomal abnormality influences the expression of other genes related with cell cycle or senescence. PMID:25472257

Song, K; Jang, G; Lee, B C

2014-12-01

52

Annexin A2 participates in human skin keloid formation by inhibiting fibroblast proliferation.  

Science.gov (United States)

Abnormal scarring results from the expression and composition of extracellular matrix molecules. The transcription and translation of collagens I and III, fibronectin, laminin, periostin, and tenascin are all increased in raised dermal scar tissue. However, human keloid development is not fully defined. In this study, we identified proteins expressed differentially between normal skin and keloid scar tissues and examined their function in keloid formation using fibroblasts. Skin specimens from normal volunteers and patients with keloids were obtained by skin biopsy. Whole proteins were isolated by two-dimensional electrophoresis, and differentially expressed proteins were identified by matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry. Protein function was determined by proliferation assay using annexin A2-overexpressing keloid fibroblasts. The expression of 11 protein spots was altered by at least 1.5-fold in patients with keloids than in normal volunteers. Of these proteins, annexin A2, a pre-serum amyloid P component, serum albumin precursor, and tryptase-I, were down-regulated in keloid tissue compared to normal skin. Collagen alpha 1(V) chain precursor, collagen alpha 1(I) chain precursor, ferritin light subunit, alpha 1(III) collagen, 6-phosphogluconolactonase, and calponin 2 were up-regulated. Diminished expression of annexin A2 was confirmed by immunoblotting and immunohistochemistry. Treatment with the recombinant human epidermal growth factor increased proliferation of keloid fibroblasts, which was more inhibited in annexin A2-overexpressing fibroblasts than in non-transfected control cells. These results imply that annexin A2 may participate in keloid formation by inhibiting keloid fibroblast proliferation. Therefore, it is concluded that annexin A2 may be a valuable therapeutic target for keloid lesions. PMID:24402284

Kim, Soon Heum; Jung, Seung-Hyo; Chung, Hong; Jo, Dong In; Kim, Cheol Keun; Park, Seung Hwa; Won, Kyung-Jong; Jeon, Hyun Soo; Kim, Bokyung

2014-05-01

53

Effects of recombinant human epidermal growth factor on the proliferation and radiation survival of human fibroblast cell lines in vitro  

Energy Technology Data Exchange (ETDEWEB)

To explore the effect of recombinant human EGF on the proliferation and survival of human fibroblast cell lines following irradiation. Fibroblast was originated human skin and primary cultured. The trypan blue stain assay and MTT assay were used to study the proliferative effects of EGF on human fibroblast cell lines in vitro. An incubation of fibroblasts with rhEGF for 24 hours immediately after irradiation was counted everyday. Cell cycle distributions were analyzed by FACS analysis. Number of fibroblast was significant more increased rhEGF (1.0 nM, 10 nM, 100 nM, 1,000 nM) treated cell than control after 8 Gy irradiation. Most effective dose of rhEGF was at 160 nM. These survival differences were maintained at 1 week later. Proportion of S phase was significantly increased on rhEGF treated cells. rhEGF cause increased fibroblast proliferation following irradiation. We expect that rhEGF was effective for radiation induced wound healing.

Kim, Hyun Sook; Kang, Ki Mun; Na, Jae Boem; Chai, Gyu Young [Gyeongsang National University College of Medicine, Jinju (Korea, Republic of); Lee, Sang Wook [University of Ulsan College of Medicine, Asan Medical Center, Seoul (Korea, Republic of)

2006-09-15

54

Effects of recombinant human epidermal growth factor on the proliferation and radiation survival of human fibroblast cell lines in vitro  

International Nuclear Information System (INIS)

To explore the effect of recombinant human EGF on the proliferation and survival of human fibroblast cell lines following irradiation. Fibroblast was originated human skin and primary cultured. The trypan blue stain assay and MTT assay were used to study the proliferative effects of EGF on human fibroblast cell lines in vitro. An incubation of fibroblasts with rhEGF for 24 hours immediately after irradiation was counted everyday. Cell cycle distributions were analyzed by FACS analysis. Number of fibroblast was significant more increased rhEGF (1.0 nM, 10 nM, 100 nM, 1,000 nM) treated cell than control after 8 Gy irradiation. Most effective dose of rhEGF was at 160 nM. These survival differences were maintained at 1 week later. Proportion of S phase was significantly increased on rhEGF treated cells. rhEGF cause increased fibroblast proliferation following irradiation. We expect that rhEGF was effective for radiation induced wound healing

55

Relationship between the in vitro radiosensitivity of skin fibroblasts and the expression of subcutaneous fibrosis, telangiectasia, and skin erythema after radiotherapy  

DEFF Research Database (Denmark)

To investigate if the occurrence of subcutaneous fibrosis after radiotherapy in an unselected group of breast cancer patients is related to cellular radiosensitivity of skin fibroblasts as measured in a clonogenic assay.

Johansen, J; Bentzen, SØren M

1996-01-01

56

Increased collagen production in fibroblasts cultured from irradiated skin and effect of TGF ?1– clinical study  

OpenAIRE

Fibrosis in normal tissues is a common and dose-limiting late complication of radiotherapy at many cancer sites, but its pathogenesis is poorly understood. We undertook a controlled study of the effect of irradiation on the collagen production of fibroblasts cultured from skin biopsies taken from patients undergoing radiotherapy treatment. Eight weeks after a single 8 Gy fraction using 300 kV X-rays, five patients treated at the Royal Marsden Hospital underwent biopsy of the irradiated site a...

Illsley, M. C.; Peacock, J. H.; Mcanulty, R. J.; Yarnold, J. R.

2000-01-01

57

Silver nanoparticles mediate differential responses in keratinocytes and fibroblasts during skin wound healing  

OpenAIRE

With advances in nanotechnology, pure silver has been recently engineered into nanometer-sized particles (diameter < 100 nm) for use in the treatment of wounds. In conjunction with other studies, we previously demonstrated that the topical application of silver nanoparticles (AgNPs) can promote wound healing through the modulation of cytokines. Nonetheless, the question as to whether AgNPs can affect various skin cell types - keratinocytes and fibroblasts - during the wound-healing process st...

Ho, Cm; Lui, Vch; Chen, Y.; Che, Cm; Tam, Pkh; Wong, Kky; Liu, X.; Lee, Py

2010-01-01

58

Senescent fibroblasts enhance early skin carcinogenic events via a paracrine MMP-PAR-1 axis.  

Science.gov (United States)

The incidence of carcinoma increases greatly with aging, but the cellular and molecular mechanisms underlying this correlation are only partly known. It is established that senescent fibroblasts promote the malignant progression of already-transformed cells through secretion of inflammatory mediators. We investigated here whether the senescent fibroblast secretome might have an impact on the very first stages of carcinogenesis. We chose the cultured normal primary human epidermal keratinocyte model, because after these cells reach the senescence plateau, cells with transformed and tumorigenic properties systematically and spontaneously emerge from the plateau. In the presence of medium conditioned by autologous senescent dermal fibroblasts, a higher frequency of post-senescence emergence was observed and the post-senescence emergent cells showed enhanced migratory properties and a more marked epithelial-mesenchymal transition. Using pharmacological inhibitors, siRNAs, and blocking antibodies, we demonstrated that the MMP-1 and MMP-2 matrix metalloproteinases, known to participate in late stages of cancer invasion and metastasis, are responsible for this enhancement of early migratory capacity. We present evidence that MMPs act by activating the protease-activated receptor 1 (PAR-1), whose expression is specifically increased in post-senescence emergent keratinocytes. The physiopathological relevance of these results was tested by analyzing MMP activity and PAR-1 expression in skin sections. Both were higher in skin sections from aged subjects than in ones from young subjects. Altogether, our results suggest that during aging, the dermal and epidermal skin compartments might be activated coordinately for initiation of skin carcinoma, via a paracrine axis in which MMPs secreted by senescent fibroblasts promote very early epithelial-mesenchymal transition of keratinocytes undergoing transformation and oversynthesizing the MMP-activatable receptor PAR-1. PMID:23675494

Malaquin, Nicolas; Vercamer, Chantal; Bouali, Fatima; Martien, Sébastien; Deruy, Emeric; Wernert, Nicolas; Chwastyniak, Maggy; Pinet, Florence; Abbadie, Corinne; Pourtier, Albin

2013-01-01

59

DNA damage in human skin fibroblasts exposed to UVA light used in clinical PUVA treatment  

International Nuclear Information System (INIS)

Human skin fibroblasts were irradiated with a clinically used UVA light source. The doses (1.1 and 3 J/cm2) were similar to those reaching the dermis during clinical PUVA treatment of psoriasis. DNA strand breaks, as determined by alkaline elution, were formed in a dose-dependent way and disappeared within 1 hr of postincubation at 37 degrees C. These findings have clinical implications since UVA-induced DNA damage may be accompanied by mutagenic and tumor promoting effects

60

Protective activity of hamamelitannin on cell damage of murine skin fibroblasts induced by UVB irradiation.  

Science.gov (United States)

The protective activities of hamamelitannin (2',5-di-O-galloyl-hamamelose) in Hamamelis virginiana L. and its related compound, gallic acid, on damaged murine skin fibroblasts induced by UVB irradiation were investigated. In order to exclude the UV absorbing effect of the compounds, the protection study was performed such that the fibroblasts were pretreated with hamamelitannin or gallic acid for 24 h before UVB irradiation. At 200 microM concentration, hamamelitannin gave the higher survival of 72.6 +/- 0.4% in comparison with that of gallic acid (35.5 +/- 1.0%), while UVB absorbers such as 2-ethylhexyl p-methoxycinnamate and hexylbenzoate did not show such protection. The scavenging activities of hamamelitannin and gallic acid against active oxygens such as superoxide anion radicals, hydroxyl radicals and singlet oxygens were evaluated using electron spin resonance (ESR-spin trapping method). Hamamelitannin and gallic acid showed potent scavenging activities against all active oxygens tested. Furthermore, the association of hamamelitannin to fibroblasts was examined by comparing it with that of gallic acid, and the following results were obtained: (1) hamamelitannin reduces the reaction rate of liposome entrapped-nitroblue tetrazolium (NBT) with external superoxide anions, and (2) several glycosides associate with fibroblasts. From these results, it was concluded that hamamelitannin protects murine fibroblasts against external active oxygens by associating with the cell surface through its sugar moiety. PMID:7577835

Masaki, H; Atsumi, T; Sakurai, H

1995-07-01

61

Transformation of Human Mesenchymal Cells and Skin Fibroblasts into Hematopoietic Cells  

Science.gov (United States)

Patients with prolonged myelosuppression require frequent platelet and occasional granulocyte transfusions. Multi-donor transfusions induce alloimmunization, thereby increasing morbidity and mortality. Therefore, an autologous or HLA-matched allogeneic source of platelets and granulocytes is needed. To determine whether nonhematopoietic cells can be reprogrammed into hematopoietic cells, human mesenchymal stromal cells (MSCs) and skin fibroblasts were incubated with the demethylating agent 5-azacytidine (Aza) and the growth factors (GF) granulocyte-macrophage colony-stimulating factor and stem cell factor. This treatment transformed MSCs to round, non-adherent cells expressing T-, B-, myeloid-, or stem/progenitor-cell markers. The transformed cells engrafted as hematopoietic cells in bone marrow of immunodeficient mice. DNA methylation and mRNA array analysis suggested that Aza and GF treatment demethylated and activated HOXB genes. Indeed, transfection of MSCs or skin fibroblasts with HOXB4, HOXB5, and HOXB2 genes transformed them into hematopoietic cells. Further studies are needed to determine whether transformed MSCs or skin fibroblasts are suitable for therapy. PMID:21731684

Harris, David M.; Hazan-Haley, Inbal; Coombes, Kevin; Bueso-Ramos, Carlos; Liu, Jie; Liu, Zhiming; Li, Ping; Ravoori, Murali; Abruzzo, Lynne; Han, Lin; Singh, Sheela; Sun, Michael; Kundra, Vikas; Kurzrock, Razelle; Estrov, Zeev

2011-01-01

62

Human amniotic fluid derived cells can competently substitute dermal fibroblasts in a tissue-engineered dermo-epidermal skin analog  

OpenAIRE

PURPOSE: Human amniotic fluid comprises cells with high differentiation capacity, thus representing a potential cell source for skin tissue engineering. In this experimental study, we investigated the ability of human amniotic fluid derived cells to substitute dermal fibroblasts and support epidermis formation and stratification in a humanized animal model. METHODS: Dermo-epidermal skin grafts with either amniocytes or with fibroblasts in the dermis were compared in a rat model. Full-thicknes...

Hartmann-fritsch, Fabienne; Hosper, Nynke; Luginbu?hl, Joachim; Biedermann, Thomas; Reichmann, Ernst; Meuli, Martin

2013-01-01

63

Resistance to 1,25-dihydroxyvitamin D. Association with heterogeneous defects in cultured skin fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

The authors evaluated the interaction of (/sub 3/H)1,25(OH)/sup 2/D3 with skin fibroblasts cultured from normal subjects or from affected members of six kindreds with rickets and resistance to 1-alpha, 25(OH)/sub 2/D (1,25(OH)/sub 2/D). They analyzed two aspects of the radioligand interaction; nuclear uptake with dispersed, intact cells at 37 degrees C and binding at 0 degrees C with soluble extract (cytosol) prepared from cells disrupted in buffer. With normal fibroblasts the affinity and capacity of nuclear uptake of (/sub 3/H)1,25(OH)/sup 2/D3 were 0.5 nM and 10,300 sites per cell, respectively; for binding with cytosol these were 0.13 nM and 8,900 sites per cell, respectively. The following four patterns of interaction with (/sub 3/H)1,25(OH)/sup 2/D3 were observed with cells cultured from affected patients. In all cases where the radioligand bound with high affinity in nucleus or cytosol, the nucleus- or cytosol-associated radioligand exhibited normal sedimentation velocity on sucrose density gradients. When two kindreds exhibited similar patterns (i.e. pattern a or c) with the analyses of cultured fibroblasts, clinical features in affected members suggested that the underlying genetic defects were not identical. In conclusion: (a) Fibroblasts cultured from human skin manifest nuclear uptake and cytosol binding of (/sub 3/H)1,25(OH)/sup 2/D3 that is an expression of the genes determining these processes in target tissues. (b) Based upon data from clinical evaluations and from analyses of cultured fibroblasts, severe resistance to 1,25(OH)/sup 2/D resulted from five or six distinct genetic mutations in six kindreds.

Liberman, U.A.; Eil, C.; Marx, S.J.

1983-02-01

64

Resistance to 1,25-dihydroxyvitamin D. Association with heterogeneous defects in cultured skin fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

We evaluated the interaction of (/sup 3/H)1,25(OH)/sub 2/D/sub 3/ with skin fibroblasts cultured from normal subjects or from affected members of six kindreds with rickets and resistance to 1-alpha, 25(OH)/sub 2/D (1,25(OH)/sub 2/D). We analyzed two aspects of the radioligand interaction; nuclear uptake with dispersed, intact cells at 37 degrees C and binding at 0 degrees C with soluble extract (cytosol) prepared from cells disrupted in buffer containing 300 mM KCl and 10 mM sodium molybdate. With normal fibroblasts the affinity and capacity of nuclear uptake of (/sup 3/H)1,25(OH)/sub 2/D/sub 3/ were 0.5 nM and 10,300 sites per cell, respectively; for binding with cytosol these were 0.13 nM and 8,900 sites per cell, respectively. In all cases where the radioligand bound with high affinity in nucleus or cytosol, the nucleus- or cytosol-associated radioligand exhibited normal sedimentation velocity on sucrose density gradients. When two kindreds exhibited similar patterns (i.e. pattern a or c) with the analyses of cultured fibroblasts, clinical features in affected members suggested that the underlying genetic defects were not identical. In conclusion: (a) Fibroblasts cultured from human skin manifest nuclear uptake and cytosol binding of (/sup 3/H)1,25(OH)/sub 2/D/sub 3/ that is an expression of the genes determining these processes in target tissues. (b) Based upon data from clinical evaluations and from analyses of cultured fibroblasts, severe resistance to 1,25(OH)/sub 2/D resulted from five or six distinct genetic mutations in six kindreds.

Liberman, U.A.; Eil, C.; Marx, S.J.

1983-02-01

65

Resistance to 1,25-dihydroxyvitamin D. Association with heterogeneous defects in cultured skin fibroblasts  

International Nuclear Information System (INIS)

We evaluated the interaction of [3H]1,25(OH)2D3 with skin fibroblasts cultured from normal subjects or from affected members of six kindreds with rickets and resistance to 1-alpha, 25(OH)2D [1,25(OH)2D]. We analyzed two aspects of the radioligand interaction; nuclear uptake with dispersed, intact cells at 37 degrees C and binding at 0 degrees C with soluble extract (cytosol) prepared from cells disrupted in buffer containing 300 mM KCl and 10 mM sodium molybdate. With normal fibroblasts the affinity and capacity of nuclear uptake of [3H]1,25(OH)2D3 were 0.5 nM and 10,300 sites per cell, respectively; for binding with cytosol these were 0.13 nM and 8,900 sites per cell, respectively. In all cases where the radioligand bound with high affinity in nucleus or cytosol, the nucleus- or cytosol-associated radioligand exhibited normal sedimentation velocity on sucrose density gradients. When two kindreds exhibited similar patterns (i.e. pattern a or c) with the analyses of cultured fibroblasts, clinical features in affected members suggested that the underlying genetic defects were not identical. In conclusion: (a) Fibroblasts cultured from human skin manifest nuclear uptake and cytosol binding of [3H]1,25(OH)2D3 that is an expression of the genes determining these processes in target tissues. (b) Based upon data from clinical evaluations and from analyses om clinical evaluations and from analyses of cultured fibroblasts, severe resistance to 1,25(OH)2D resulted from five or six distinct genetic mutations in six kindreds

66

Resistance to 1,25-dihydroxyvitamin D. Association with heterogeneous defects in cultured skin fibroblasts  

International Nuclear Information System (INIS)

The authors evaluated the interaction of [3H]1,25(OH)2D3 with skin fibroblasts cultured from normal subjects or from affected members of six kindreds with rickets and resistance to 1-alpha, 25(OH)2D [1,25(OH)2D]. They analyzed two aspects of the radioligand interaction; nuclear uptake with dispersed, intact cells at 37 degrees C and binding at 0 degrees C with soluble extract (cytosol) prepared from cells disrupted in buffer. With normal fibroblasts the affinity and capacity of nuclear uptake of [3H]1,25(OH)2D3 were 0.5 nM and 10,300 sites per cell, respectively; for binding with cytosol these were 0.13 nM and 8,900 sites per cell, respectively. The following four patterns of interaction with [3H]1,25(OH)2D3 were observed with cells cultured from affected patients. In all cases where the radioligand bound with high affinity in nucleus or cytosol, the nucleus- or cytosol-associated radioligand exhibited normal sedimentation velocity on sucrose density gradients. When two kindreds exhibited similar patterns (i.e. pattern a or c) with the analyses of cultured fibroblasts, clinical features in affected members suggested that the underlying genetic defects were not identical. In conclusion: (a) Fibroblasts cultured from human skin manifest nuclear uptake and cytosol binding of [3H]1,25(OH)2D3 that is an expression of the genes determining these processes in target nes determining these processes in target tissues. (b) Based upon data from clinical evaluations and from analyses of cultured fibroblasts, severe resistance to 1,25(OH)2D resulted from five or six distinct genetic mutations in six kindreds

67

Abnormal phenotype of cultured fibroblasts in human skin with chronic radiotherapy damage  

International Nuclear Information System (INIS)

Purpose: The pathophysiological aspects of radiation-induced fibrosis (RIF) have not been well characterized. We therefore cultured human fibroblasts from samples of skin with RIF to investigate the long-term effects of therapeutic irradiation. Materials and methods: Biopsies of normal and RIF skin were obtained from patients previously irradiated for cancer, without recurrence. Cells were extracted from dermis samples by the outgrowth technique, seeded as monolayers and cultured at confluence. Enzyme activities and proteins were assayed, RNA was isolated and Northern blot analysis was performed on surviving cells between passages 2 and 5. Results: RIF cell cultures displayed heterogeneous fibroblasts populations. The initial outgrowth consisted of one-third small cells that floated rapidly, one-third spindle-shaped cells migrating far from the explant to form islets and one-third large pleiomorphic cells. In subsequent subcultures, surviving cells exhibited either myofibroblastic characteristics with a normal proliferative capacity or senescent morphology with a reduced proliferative capacity. These RIF cells had a brief finite lifespan, with dramatically reduced growth rate during their initial outgrowth and the following passages. Study of the antioxidant metabolism showed that Mn superoxide dismutase and catalase activities were significantly weaker in surviving RIF cells than healthy fibroblasts. These exhausted RIF cells exhibited no overexpression of transformi exhibited no overexpression of transforming growth factor ? or tissue inhibitor of metalloproteinase. Conclusion: Irradiation may lead to apparently contradictory effects such as fibrosis and necrosis in clinical practice. In cell culture, we observed two main cellular phenotypes which may be related to both processes, i.e. myofibroblast-like cells and fibrocyte-like cells. These two phenotypes may represent two steps in the differentiation induced as a long-term effect of therapeutic irradiation of the skin. Cell culture probably accelerates the induction of the terminal differentiation in RIF fibroblasts. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

68

MUC1 Is Expressed by Human Skin Fibroblasts and Plays a Role in Cell Adhesion and Migration.  

Science.gov (United States)

The mucin MUC1 is expressed by normal and cancerous epithelial cells and some nonepithelial cells in which it plays roles in regulating adhesion, migration, and cell signaling. In the present studies we found that MUC1 is expressed by normal human neonatal and adult skin fibroblasts. Fibroblasts are usually considered negative for MUC1 expression. Reverse-transcription polymerase chain reaction and Western blot analyses indicate the presence of full-length MUC1, and immunofluorescence and subcellular fractionation studies show that the protein is expressed on the plasma membrane. Immunohistochemical analyses confirmed the expression of MUC1 by fibroblasts in cryosections of normal human skin. Silencing MUC1 expression in fibroblasts using MUC1 shRNA increased the adhesion of cells to collagen and laminin. Transfection with MUC1 shRNA also increased fibroblast migration on collagen as measured in a wound-healing assay. The expression of ?2-integrin was increased in MUC1 shRNA-transfected fibroblasts in which it was localized to membrane ruffles, providing a possible explanation for the increased cell migration on collagen. These results extend the range of expression of MUC1 to skin fibroblasts and suggest a functional role for MUC1 in fibroblast adhesion and motility. PMID:24804164

Kumar, Priyadarsini; Ji, Jennifer; Thirkill, Twanda L; Douglas, Gordon C

2014-04-01

69

Absence of ?-galactosidase cross-correction in Fabry heterozygote cultured skin fibroblasts.  

Science.gov (United States)

Fabry disease (FD) is an X-linked lysosomal storage disorder resulting from deficiency of ?-galactosidase A (GLA). Traditionally, heterozygotes were considered asymptomatic carriers of FD, but it is now apparent that the asymptomatic female carrier is the exception and most heterozygotes suffer significant multisystemic disease. To determine why the process of cross-correction does not occur effectively in FD heterozygotes, we investigated GLA production and secretion in cultured skin fibroblasts as well as GLA levels in plasma. The maturation of GLA was similar in FD heterozygotes and control fibroblasts, confirming that both produce the 46kDa mature form; the same as that present in control plasma. However, the proportion of GLA secreted into the culture media was substantially less than eight other lysosomal proteins. Artificial generation of FD heterozygotes in cellulo, along with another lysosomal storage disorder, mucopolysaccharidosis type II, revealed no cross-correction in the FD system, whereas MPS II fibroblasts were able to cross-correct. In plasma, GLA was present as the 46kDa mature form, which lacks the mannose 6-phosphorylated moiety and is not able to be efficiently endocytosed by affected cells. Our evidence shows that fibroblasts secrete minimal amounts of GLA and consequently normal fibroblasts are unable to cross-correct FD fibroblasts. We suggest that symptomatic FD heterozygotes arise due to the secretion of primarily the mature form, with only small amounts of the mannose 6-phosphorylated form of GLA from unaffected cells. This limits capacity for enzyme cross correction of affected cells, despite uptake of exogenous recombinant GLA. PMID:25468650

Fuller, Maria; Mellett, Natalie; Hein, Leanne K; Brooks, Doug A; Meikle, Peter J

2015-02-01

70

Chitosan scaffold co-cultured with keratinocyte and fibroblast heals full thickness skin wounds in rabbit.  

Science.gov (United States)

This study evaluated the modulatory effect of chitosan sponge co-cultured with keratinocyte and fibroblast on wound healing. Dermal fibroblasts and keratinocyte isolated from rabbit skin were co-cultured on chitosan sponge, to fabricate cell-loaded chitosan tissue engineered construct. Full thickness excision wounds created on the rabbit dorsum were treated with three types of graft materials – a noncellular chitosan graft, homologous keratinocyte fibroblast loaded chitosan, and a commercial product. Postgraft skin-wound samples were examined histomorphologically at 7th, 14th, and 28th day after staining with hematoxylin and eosin, picrosirius red and/or immunohistochemistry. Wound healing parameters considered were the extent of re-epithelialization, collagen deposition, and neoangiogenesis. The number of proliferating cells, vimentin positive cells, and alpha smooth muscle actin cells were also quantified. The histology results suggested that the grafts aided wound healing but, the cell-loaded graft induced a differential pattern of healing and had lower scarring tendency. The cell-loaded tissue construct may be useful as a therapeutic graft for treating wounds where there is a total loss of tissue and cells as in burn injury. PMID:24133040

Revi, Deepa; Paul, Willi; Anilkumar, T V; Sharma, Chandra P

2014-09-01

71

Biosynthetic support based on dendritic poly(L-lysine) improves human skin fibroblasts attachment.  

Science.gov (United States)

Poly(L-lysine) (PLL) dendrigrafts (DGLs) are arborescent biosynthetic polymers of regular and controlled structures. They have specific properties such as biocompatibility and non-immunogenicity, and their surface density of NH2 functions can be easily modified and therefore appears as a powerful tool for the functionalization of hydrophobic polymers used in the context of tissue engineering. In this study, we evaluated several criteria of human skin fibroblasts when cultured with DGL of generations 2, 3 and 4, with linear PLL polymer as reference. In aqueous phase, DGLs and PLL displayed a similar cytotoxicity towards fibroblasts. Plastic culture plates grafted with DGLs were further characterized as homogeneous surfaces by atomic force microscopy and surface characterization by amino density estimation by colorimetric assay. Proliferation of fibroblasts was increased when cultured onto PLL and DGLs monolayers when compared with crude plates. Cellular adhesion was increased by 20% on DGLs in comparison to PLL. Integrin ?5 subunit protein expression level was increased after 48 h of culture on DGLs, in comparison to control or PLL-coated surfaces. The presence of DGLs did not lead to overexpression or activation of matrix metalloproteinases 2 and 9. Finally, fibroblasts adhesion was increased by 40% on poly-(lactic-co-glycolic acid) matrices functionalized with DGLs when compared to PLL. Overall, these features make DGL promising candidates for the surface engineering of biomaterials in tissue engineering. PMID:24116875

Lorion, Chloé; Faye, Clément; Maret, Barbara; Trimaille, Thomas; Régnier, Thomas; Sommer, Pascal; Debret, Romain

2014-01-01

72

Differentiation state of skin fibroblast cultures versus risk of subcutaneous fibrosis after radiotherapy  

International Nuclear Information System (INIS)

Background and purpose: There is increasing evidence for patient-to-patient variation in the response of normal tissue to radiotherapy. Recently, it has been suggested that accumulation of functional fibrocytes may be a key step in the development of radiation-induced fibrosis. Therefore, we have examined a possible relationship between the differentiation state of untreated fibroblasts and the risk of radiation-induced subcutaneous fibrosis in individual patients. Materials and methods: We used skin fibroblast cultures isolated from eight postmastectomy radiotherapy patients whose individual clinical radiosensitivity was assessed by the mean excess risk of fibrosis. Different types of potentially mitotic progenitor fibroblasts (MF) and postmitotic functional fibrocytes (PMF) in the terminal differentiation lineage (MFI approaches MFII approaches MFIII approaches PMF) were scored morphologically in clonal culture. Progression of differentiation was quantified by the ratio L/E of colony-forming late (MFIII and late MFII) and early (MFI and early MFII) progenitors. Results: We observed a correlation between the ratio L/E and the mean risk of fibrosis (rS=0.743, P=0.03), indicating an approximately 10-fold increase in L/E with an increasing risk of fibrosis. This was paralleled by a decreasing trend in the absolute numbers of early progenitor types. By contrast, there was no significant correlation between the plating efficiency and the risk of fibrosis. Conclefficiency and the risk of fibrosis. Conclusions: The data suggest that the risk of fibrosis increases with the progression of the differentiation of untreated progenitor fibroblasts, indicating that the progression of fibroblast differentiation may be a co-factor in the development of radiation-induced fibrosis. If this hypothesis is validated, it provides a rationale for a novel predictive test to identify patients with an increased risk of subcutaneous fibrosis. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

73

Relationship between DNA double-strand breaks, cell killing, and fibrosis studied in confluent skin fibroblasts derived from breast cancer patients  

DEFF Research Database (Denmark)

To investigate the relationship between DNA double-strand breaks (dsbs), cell killing, and fibrosis using skin fibroblasts derived from breast cancer patients who received postmastectomy radiotherapy.

Dikomey, E; Brammer, I

2000-01-01

74

Fibroblast radiosensitivity versus acute and late normal skin responses in patients treated for breast cancer  

International Nuclear Information System (INIS)

Purpose/Objective: To determine if the radiosensitivity of normal human skin fibroblasts, measured in early passage cultures, is significantly correlated with the degree of acute or late normal skin damage in patients treated for breast cancer with radiotherapy. Methods and Materials: In the 1970s, a series of breast cancer patients was treated at the Department of Oncology in Gothenburg, Sweden with postoperative irradiation to the parasternal region. Patients were treated bilaterally using different fractionation schedules and doses to the right and left fields. Peak acute reactions were scored on a six-point scale, and skin erythema was measured by reflectance spectrophotometry. Telangiectasia was graded over time on a six-point scale. In April 1992, two small skin biopsies were obtained from 22 patients in two treatment groups (i.e., four dose-fractionation schedules) and, using either delayed or immediate plating, fibroblast radiosensitivity was measured in early passage cultures by clonogenic survival, after high and low dose-rate irradiations. Survival at 2.0 Gy (SF2) was calculated from complete survival curves. Results: To test assay reproducibility, SF2 values derived from paired biopsies of the same patient (12 cases) were compared. A reasonably good correlation (p = 0.075) was obtained for SF2s determined by high dose-rate irradiations with immediate plating, but not for delayed plating or low dose-rate treatments. The median coefficient of variation in th The median coefficient of variation in the replicate SF2s after high dose-rate treatment and immediate plating was 13%, suggesting that the poor correlation in paired SF2 values is due to the magnitude of the uncertainty in SF2 relative to the overall spread in SF2 values between patients (CV = 28%). Individual SF2 values and averaged values from patients with data from two biopsies were compared with the acute and late clinical reactions. A significant negative correlation was found between SF2 and relative clinical response, but only when averaged high dose-rate SF2 values and telangiectasia scores were compared. There was no significant correlation between average SF2 values and acute responses or between individual SF2 measurements and either the acute or late clinical response. Conclusion: The results of this study suggest that the degree of late telangiectasia is at least partially dependent upon the intrinsic cellular radiosensitivity of normal fibroblasts, but the relationship is not clear cut. Multiple replicate assays are necessary to obtain reliable estimates of fibroblast SF2 values using current techniques

75

Binding of plasma fibronectin to cell layers of human skin fibroblasts  

OpenAIRE

Human plasma fibronectin bound to confluent cell layers of cultured human-skin fibroblasts in two distinct pools. Initial binding of fibronectin occurred in a deoxycholate-soluble pool (Pool I). Binding in Pool I was reversible and reached a steady state after 3 h. After longer periods of incubation, fibronectin became bound in a deoxycholate-insoluble pool (Pool II). Binding in Pool II was irreversible and proceeded at a linear rate for 30 h. After 30 h of incubation, a significant proportio...

1983-01-01

76

Doxorubicin-induced inhibition of prolyl hydroxylation during collagen biosynthesis in human skin fibroblast cultures. Relevance to imparied wound healing.  

OpenAIRE

Previous clinical and experimental observations have indicated that wound healing is impaired as a result of treatment with doxorubicin, a chemotherapeutic agent. In this study, the effects of doxorubicin were examined in human skin fibroblast cultures with respect to collagen production and fibroblast proliferation. The results indicated that the synthesis of hydroxyproline as a marker of collagen production was markedly reduced, with an approximate concentration of inhibitor yielding 50% in...

Sasaki, T.; Holeyfield, K. C.; Uitto, J.

1987-01-01

77

Analysis of amyloid precursor protein mRNAs in skin fibroblasts in Down's syndrome.  

Science.gov (United States)

We examined amyloid precursor protein (APP) mRNAs expression in skin fibroblasts from Down's syndrome (DS) patients of different ages to determine the time of occurrence of abnormal splicing. The ratio of APP770 + 751 mRNA to APP695 mRNA (APP770 + 751/695) was significantly increased in the young DS group and adult DS group compared with the age-matched control groups (p < 0.01, p < 0.05), but no significant increase was observed in the aged DS group compared with the age-matched control group. These findings suggest that metabolic abnormalities of the APP gene occur at a very early stage of DS, at a mean age of about 5 years. Therefore, metabolic abnormalities of the APP gene are considered to appear at a very young age also Alzheimer's disease (AD). In this study, we confirmed that examination of the APP gene in skin fibroblasts might be useful for early diagnosis of AD. PMID:8866680

Urakami, K; Kataoka, J; Okada, A; Isoe, K; Wakutani, Y; Ji, Y; Adachi, Y; Ohno, K; Takahashi, K

1996-01-01

78

Zinc and propolis reduces cytotoxicity and proliferation in skin fibroblast cell culture: total polyphenol content and antioxidant capacity of propolis.  

Science.gov (United States)

It has been demonstrated that zinc exerts its beneficial influence on skin fibroblasts. Propolis, a complex mixture of plant-derived and bees' products, was reported to stimulate cicatrization processes in skin and prevent infections. The aim of this study was to find out how zinc and propolis influence human skin fibroblasts in cell culture and to compare the effect of individual compounds to the effect of a mixture of zinc and propolis. In this study, zinc, as zinc aspartate, at a concentration of 16 ?M, increased human fibroblasts proliferation in cell culture, whereas propolis at a concentration of 0.01% (w/v) revealed antiproliferative and cytotoxic action followed by mild cell necrosis. In culture, zinc was effectively transported into fibroblasts, and propolis inhibited the amount of zinc incorporated into the cells. An addition of propolis to the medium caused a decrease in the Zn(II) amount incorporated into fibroblasts. The obtained results also indicate an appreciable antioxidant property of propolis and revealed its potential as a supplement when applied at doses lower than 0.01% (w/v). In conclusion, the present study showed that zinc had a protective effect on human cultured fibroblasts' viability, although propolis revealed its antiproliferative action and caused mild necrosis. PMID:24913100

Tyszka-Czochara, Ma?gorzata; Pa?ko, Pawe?; Reczy?ski, Witold; Szlósarczyk, Marek; Bystrowska, Beata; Opoka, W?odzimierz

2014-07-01

79

Protein, RNA, and DNA synthesis in cultures of skin fibroblasts from healthy subjects and patients with rheumatic diseases  

International Nuclear Information System (INIS)

To study the mechanism of the lasting disturbance of fibroblast function, protein, RNA and DNA synthesis was investigated in skin fibroblasts from patients with rheumatoid arthritis (RA) and systemic scleroderma (SS). The labeled precursors used to analyze synthesis of protein, RNA, and DNA were 14C-protein hydrolysate, (14C)uridine, and (14C) thymidine. Stimulation was determined by measuring incorporation of (14C)proline into fibroblast proteins. During analysis of stability of fast-labeled RNA tests were carried out to discover whether all measurable radioactivity belonged to RNA molecules

80

Gallic Acid Regulates Skin Photoaging in UVB-exposed Fibroblast and Hairless Mice.  

Science.gov (United States)

Ultraviolet (UV) radiation is the primary factor in skin photoaging, which is characterized by wrinkle formation, dryness, and thickening. The mechanisms underlying skin photoaging are closely associated with degradation of collagen via upregulation of matrix metalloproteinase (MMP) activity, which is induced by reactive oxygen species (ROS) production. Gallic acid (GA), a phenolic compound, possesses a variety of biological activities including antioxidant and antiinflammatory activities. We investigated the protective effects of GA against photoaging caused by UVB irradiation using normal human dermal fibroblasts (NHDFs) in vitro and hairless mice in vivo. The production levels of ROS, interlukin-6, and MMP-1 were significantly suppressed, and type I procollagen expression was stimulated in UVB-irradiated and GA-treated NHDFs. GA treatment inhibited the activity of transcription factor activation protein 1. The effects of GA following topical application and dietary administration were examined by measuring wrinkle formation, histological modification, protein expression, and physiological changes such as stratum corneum hydration, transepidermal water loss, and erythema index. We found that GA decreased dryness, skin thickness, and wrinkle formation via negative modulation of MMP-1 secretion and positive regulation of elastin, type I procollagen, and transforming growth factor-?1. Our data indicate that GA is a potential candidate for the prevention of UVB-induced premature skin aging. Copyright © 2014 John Wiley & Sons, Ltd. PMID:25131997

Hwang, Eunson; Park, Sang-Yong; Lee, Hyun Ji; Lee, Tae Youp; Sun, Zheng-Wang; Yi, Tae Hoo

2014-12-01

81

Skin fibroblasts from a D-deletion type retinoblastoma patient are abnormally X-ray sensitive  

International Nuclear Information System (INIS)

Retinoblastoma is a rare malignant eye tumour that appears either spontaneously or in genetically predisposed persons. The latter group is composed of persons who inherit the tumour with a dominant mode of transmission (the familial type) and those who have a deletion in the long arm of chromosome 13 referred to as the D-deletion type. When this deletion is present it is observed in many somatic cells and is often associated with structural defects. Survivors of the genetic forms of retinoblastoma have an increased risk of the development of cancers at other sites. A single genetic locus is unlikely to predispose many somatic cells to tumour formation unless a fundamental molecular defect, possibly related to DNA repair, is present. In order to investigate this hypothesis a study was made of the in vitro X-ray sensitivity of skin fibroblasts derived from three retinoblastoma patients, comprising a pair of twins with the familial type accompanied by no gross chromosome abnormalities, and a patient with the D-deletion type. It was found that fibroblasts derived from the D-deletion patient were significantly more radiosensitive than those from the other two patients. X-ray survival curves are shown. It is concluded that skin fibroblasts derived from a patient with the D-deletion variant of retinoblastoma are abnormally radiosensitive. Future investigations may indicate a specific defect in molecular repair of DNA that will explain the predisposition of these patients to n the predisposition of these patients to the development of other tumours. (U.K.)

82

Fibroblast radiosensitivity versus acute and late normal skin responses in patients treated for breast cancer  

International Nuclear Information System (INIS)

To determine if the radiosensitivity of normal human skin fibroblasts, measured in early passage cultures, is significantly correlated with the degree of acute or late normal skin damage in patients treated for breast cancer with radiotherapy. To test assay reproducibility, SF2 values derived from paired biopsies of the same patient (12 cases) were compared. A reasonably good correlation (p = 0.075) was obtained for SF2s determined by high dose-rate irradiations with immediated plating, but not for delayed plating or low dose-rate treatments. The median coefficient of variation in the replicate SF2s after high dose-rate treatment and immediate plating was 13%, suggesting that the poor correlation in paired SF2 values is due to the magnitude of the uncertainty in SF2 relative to the overall spread in SF2 values between patients (CV = 28%). Individual SF2 values and averaged values from patients with data from two biopsies were compared with the acute and late clinical reactions. A significant negative correlation was found between SF2 and relative clinical response, but only when averaged high dose-rate SF2 values and telangiectasia scores were compared. There was no significant correlation between average SF2 values and acute responses or between individual SF2 measurements and either the acute or late clinical response. The results of this study suggest that the degree of late telangiectasia is at least partially dependent upon the intrinsic cellular radiosensitivityon the intrinsic cellular radiosensitivity of normal fibroblasts, but the relationship is not clear cut. Multiple replicate assays are necessary to obtain reliable estimates of fibroblast SF2 values using current techniques. 20 refs., 3 figs., 3 tabs

83

Ionizing radiation induces early, sustained increases in collagen biosynthesis: a 48-week study in mouse skin and skin fibroblast cultures  

International Nuclear Information System (INIS)

Groups of 10 CF1 female mice, irradiated to the thorax with a dual-head 137Cs gamma-RAY source, received single doses of 0, 5, 10, 15, or 25 Gy. One to forty-eight weeks later collagen synthesis was measured in minced skin specimens incubated in medium containing [3H]proline and then assayed for radioactive hydroxyproline. A progressive, generally dose-dependent increase in collagen biosynthesis, up to 50% above control sites, was found 1, 4, and 12 weeks after radiation exposure. These changes showed further small fluctuations at 12-36 weeks, increasing again at the 48-week interval. At the same times throughout the study fibroblasts were cultured from skin explants. Following the second subculture, these cells were also incubated in medium containing [3H]proline, and collagen synthesis was again determined by [3H]hydroxyproline assay. At all radiation dose levels studied, collagen production increased threefold by 12 weeks postradiation and remained elevated for the 48-week duration of the study. In vitro radiation dose response differences were not observed

84

Endothelin receptor antagonists: effects on extracellular matrix synthesis in primary cultures of skin fibroblasts from systemic sclerosis patients  

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Full Text Available Endothelin-1 (ET-1 seems to enhance the pro-fibrotic protein synthesis by skin fibroblasts and its effects are mediated by endothelin-A and B (ETA and ETB receptors. This study aimed to investigate the effects of ETA and ETB receptor antagonists (ETARA-sitaxentan and ETA/BRA-bosentan on type I collagen (COL-1, fibronectin (FN and fibrillin-1 (FBL-1 synthesis in primary cultures of skin fibroblasts from systemic sclerosis patients. Primary cultures of fibroblasts were obtained from skin biopsies of 6 female systemic sclerosis patients and were treated with ET-1 (100 nM for 24 and 48 hrs with or without pre-treatment (1 hr with ETARA (2 ?M or ETA/BRA (10 ?M. Primary culture of human scleroderma skin fibroblasts not treated with ET-1 or ET receptor antagonists (ETARA and ETA/BRA were used as controls. COL-1, FN and FBL-1 synthesis was evaluated by immunocytochemistry and Western blot analysis. Immunocytochemistry and Western blot analysis showed that ET-1 significantly increased COL-1 and FN synthesis at 24 and 48 hrs and FBL-1 synthesis at 48 hrs vs untreated cells. ETARA significantly contrasted the ET-1-mediated increase in COL-1 and FN at 24 hrs as well as COL-1 and FBL-1 at 48 hrs, but not FN synthesis vs ET-1-treated fibroblasts. Conversely, ETA/BRA significantly antagonized the ET-1-mediated overproduction of COL-1 and FN both at 24 and 48 hrs and the FBL-1 synthesis at 48 hrs vs ET-1-treated cells. The single ETARA treatment seems to contrast significantly the increase in COL-1 synthesis, whereas the dual ETA/BRA treatment seems active in significantly antagonizing both COL-1 and FN overproduction induced by ET-1. In conclusion, ET-1 antagonism might have positive effects in contrasting the profibrotic activity of systemic sclerosis skin fibroblasts.

B. Villaggio

2012-12-01

85

Effect of basic fibroblast growth factor-saporin mitotoxin on breast cancer cell lines in vitro.  

Science.gov (United States)

Fibroblast growth factor receptors are widely expressed on breast cancer cells and we report a preliminary study to determine whether these could be useful as potential targets for delivery of a cytotoxic fibroblast growth factor 2-saporin conjugate. We show that this mitotoxin conjugate can displace I-125-fibroblast growth factor 2 binding though with reduced affinity compared to unlabeled fibroblast growth factor 2. For 4 out of 5 cell lines it is an effective inhibitor of cell growth, and cytotoxic for at least 2 of the lines. Inhibitory effects did not depend on responsiveness of cells to fibroblast growth factor 2. This activity was not achieved with free saporin. There may be potential uses for this conjugate in both experimental systems to study receptor function and subsequent processing, and also in clinical settings to eliminate breast cancer cells. PMID:21590072

Luqmani, Y; Chander, S; Coope, R; Gomm, J; Lappi, D; Coombes, R

1997-01-01

86

Mesenchymal stem cell-conditioned medium accelerates skin wound healing: An in vitro study of fibroblast and keratinocyte scratch assays  

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We have used in vitro scratch assays to examine the relative contribution of dermal fibroblasts and keratinocytes in the wound repair process and to test the influence of mesenchymal stem cell (MSC) secreted factors on both skin cell types. Scratch assays were established using single cell and co-cultures of L929 fibroblasts and HaCaT keratinocytes, with wound closure monitored via time-lapse microscopy. Both in serum supplemented and serum free conditions, wound closure was faster in L929 fibroblast than HaCaT keratinocyte scratch assays, and in co-culture the L929 fibroblasts lead the way in closing the scratches. MSC-CM generated under serum free conditions significantly enhanced the wound closure rate of both skin cell types separately and in co-culture, whereas conditioned medium from L929 or HaCaT cultures had no significant effect. This enhancement of wound closure in the presence of MSC-CM was due to accelerated cell migration rather than increased cell proliferation. A number of wound healing mediators were identified in MSC-CM, including TGF-{beta}1, the chemokines IL-6, IL-8, MCP-1 and RANTES, and collagen type I, fibronectin, SPARC and IGFBP-7. This study suggests that the trophic activity of MSC may play a role in skin wound closure by affecting both dermal fibroblast and keratinocyte migration, along with a contribution to the formation of extracellular matrix.

Walter, M.N.M. [Institute for Science and Technology in Medicine, Keele University RJAH Orthopaedic Hospital, Oswestry, SY10 7AG (United Kingdom); School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ (United Kingdom); Wright, K.T.; Fuller, H.R. [Institute for Science and Technology in Medicine, Keele University RJAH Orthopaedic Hospital, Oswestry, SY10 7AG (United Kingdom); MacNeil, S. [Kroto Research Institute and Centre for Nanoscience and Technology, Sheffield University, Sheffield, S1 2UE (United Kingdom); Johnson, W.E.B., E-mail: w.e.johnson@aston.ac.uk [School of Life and Health Science, Aston University, Aston Triangle, Birmingham, B4 7EJ (United Kingdom)

2010-04-15

87

Cadherin-11 Induces Rheumatoid Arthritis Fibroblast-Like Synoviocytes to Form Lining Layers in Vitro  

OpenAIRE

The synovial lining of diarthrodial joints is composed of a condensed network of synoviocytes that form an intact layer via cell-to-cell contacts with significant intercellular matrix spaces. However, the molecular basis for synovial lining formation and its structural integrity has not been previously defined. In this study, using a three-dimensional fibroblast-like synoviocyte in vitro organ culture system, we provide evidence that cadherin-11 expressed in fibroblast-like synoviocytes plays...

Kiener, Hans P.; Lee, David M.; Agarwal, Sandeep K.; Brenner, Michael B.

2006-01-01

88

Cytogenetic study of skin fibroblasts in a case of accidental acute irradiation  

International Nuclear Information System (INIS)

The cytogenetic study of skin fibroblasts from a young boy, heavily irradiated by handling of an iridium-192 source of 25 curies is reported. About half of the cells examined had chromosomal abnormalities. The same clone, with multiple chromosome rearrangement, was observed in cultures from biopsies obtained 25 and 35 months after the accident. Several other clones were detected in vitro. The results obtained from cultures of biopsies from different locations show that no direct relationships were found between the absorbed dose and the frequency of stable chromosomal rearrangements. The comparison of the intrachromosomal rearrangements, mostly inversions, observed in this study with those detected in human pathology, in irradiation experiments in vitro, and in various species of primates indicates that these rearrangements do not occur at random. (orig.)

89

Multilineage differentiation potential of fibroblast-like stromal cells derived from human skin.  

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Adult stem cells that reside in adult tissues have been deemed to possess great potential for clinical application because of their capabilities of self-renewal and differentiation. However, the limitations such as infection risks and low isolation rate make the search for appropriate source to be continued. Here, we demonstrate isolation of progenitors from human foreskin tissue samples, which have fibroblast-like morphology and could be easily propagated for more than 50 passages. These foreskin-derived fibroblast-like stromal cells (FDSCs) expressed CD90, CD105, CD166, CD73, SH3, and SH4, which is similar to the immunophenotypes of human bone marrow-derived mesenchymal stem cells. In comparison with Hs68, the human fibroblast cell line, FDSCs are positive for CD105 and absent for CD54 expression. Further, FDSCs could be induced to differentiate into osteocytes, adipocytes, neural cells, smooth muscle cells, Schwann-like cells, and hepatocyte-like cells. Interestingly, when cultured in Dulbecco's modified Eagle's medium/F12 medium, FDSCs can form spheres with increased expression levels of fibronectin and vimentin. In conclusion, foreskin can serve as a valuable source of multipotent cells with the capabilities for endodermal, mesodermal, and ectodermal cells. Coupled with the advantages of their easy access, isolation, and propagation, these foreskin-derived stromal cells might be of potential use in future studies in stem cell differentiation and therapeutic application. PMID:20001268

Huang, Hsing-I; Chen, Shao-Kuan; Ling, Qing-Dong; Chien, Chih-Cheng; Liu, Hsiao-Tung; Chan, Shu-Hui

2010-05-01

90

Close-packed vesicles for diclofenac skin delivery and fibroblast targeting.  

Science.gov (United States)

Concentrated and interconnected penetration enhancer containing vesicles (PEVs) are proposed as carriers for dermal delivery of diclofenac. PEVs were prepared by using a commercial phosphatidylcholine mixture (180 mg/m) and transcutol in different amounts. Conventional liposomes were also prepared and tested as control. All vesicles showed a mean size ranging from 75 to 253 nm with fairly narrow size distribution, negative zeta potential value, and drug loading capacity between 48 and 70%. SWAXS studies showed that composition affected vesicle structure and morphology: 10 and 30% transcutol PEVs were unilamellar while liposomes and 20% transcutol PEVs were multilamellar. Rheological studies demonstrated that control liposomes and 10 and 30% transcutol containing PEVs behaved as Newtonian fluids while 20% transcutol containing PEVs showed a plastic behavior. Ex vivo (trans)dermal delivery experiments showed an improved skin deposition of diclofenac when PEVs were used. Vesicle toxicity and uptake of fibroblasts, target of inflammation treatment, were evaluated by MTT test and fluorescence microscopy. Control liposomes and PEVs were both able to interact and being internalized by the 3T3 fibroblasts at all time exposure tested. Furthermore, PEVs showed to be able to reduce the in vitro drug toxicity. PMID:23907049

Manca, Maria Letizia; Manconi, Maria; Falchi, Angela Maria; Castangia, Ines; Valenti, Donatella; Lampis, Sandrina; Fadda, Anna Maria

2013-11-01

91

Establishment, Characterization, and Toxicological Application of Loggerhead Sea Turtle (Caretta caretta) Primary Skin Fibroblast Cell Cultures.  

Science.gov (United States)

Pollution is a well-known threat to sea turtles but its impact is poorly understood. In vitro toxicity testing presents a promising avenue to assess and monitor the effects of environmental pollutants in these animals within the legal constraints of their endangered status. Reptilian cell cultures are rare and, in sea turtles, largely derived from animals affected by tumors. Here we describe the full characterization of primary skin fibroblast cell cultures derived from biopsies of multiple healthy loggerhead sea turtles (Caretta caretta), and the subsequent optimization of traditional in vitro toxicity assays to reptilian cells. Characterization included validating fibroblast cells by morphology and immunocytochemistry, and optimizing culture conditions by use of growth curve assays with a fractional factorial experimental design. Two cell viability assays, MTT and lactate dehydrogenase (LDH), and an assay measuring cytochrome P4501A (CYP1A) expression by quantitative PCR were optimized in the characterized cells. MTT and LDH assays confirmed cytotoxicity of perfluorooctanoic acid at 500 ?M following 72 and 96 h exposures while CYP1A5 induction was detected after 72 h exposure to 0.1-10 ?M benzo[a]pyrene. This research demonstrates the validity of in vitro toxicity testing in sea turtles and highlights the need to optimize mammalian assays to reptilian cells. PMID:25384208

Webb, Sarah J; Zychowski, Gregory V; Bauman, Sandy W; Higgins, Benjamin M; Raudsepp, Terje; Gollahon, Lauren S; Wooten, Kimberly J; Cole, Jennifer M; Godard-Codding, Céline

2014-12-16

92

Cultured skin keratinocytes from both normal individuals and basal cell naevus syndrome patients are more resistant to ?-rays and UV light compared with cultured skin fibroblasts  

International Nuclear Information System (INIS)

Measurement of colony-forming ability following exposure to ?-rays was performed on cultured skin keratinocytes and skin fibroblasts from normal individuals, basal cell naevus syndrome patients (BCNS) and ataxia telangiectasia (A-T) patients. The most striking observation was the radiation resistance of 8/8 keratinocyte strains compared with fibroblasts whether from BCNS patients or normals. The single A-T keratinocyte cell strain showed the same radiosensitivity as A-T fibroblast cell strains. The differential survival of keratinocytes and fibroblasts was also observed following exposure to 254 nm UV light. Survival curves of SV40 immortalized keratinocytes and retinoblasts derived from normal individuals or BCNS patients showed large shoulder regions following exposure to ?-rays or 254 nm UV light. An increased D37 rather than an increased Do was therefore the feature of such curves, contrasting with SV40 immortalized A-T keratinocytes or fibroblasts which showed a minimal shoulder effect and an increased Do. No difference in survival was observed between BCNS and normal cells following exposure to either UV or ?-rays. (author)

93

Melanin potentiates gentamicin-induced inhibition of collagen biosynthesis in human skin fibroblasts.  

Science.gov (United States)

One of the recognized side effects of gentamicin is ototoxicity. The mechanism underlying the organ specificity of this side effect of gentamicin has not been fully established. In view of the fact that a number of pharmacologic agents are known to form complexes with melanin and melanin is an abundant constituent of the inner ear tissues, we determined whether gentamicin interacts with melanin and how this process affects the biosynthesis of collagen in cultured human skin fibroblasts. Our results indicate that gentamicin forms complexes with melanin. The amount of gentamicin bound to melanin increases with increasing of initial drug concentration. The Scatchard plot analysis of drug binding to melanin showed that at least two classes of independent binding sites are implicated in gentamicin-melanin complex formation: one class with an association constant K(1) approximately 4 x 10(3) M(-1), and the second class with an association constant K(2) approximately 3 x 10(2) M(-1). The number of total binding sites (n(1)+n(2)) was calculated as about 1.36 micromol gentamicin per 1 mg melanin. We have suggested that prolidase, an enzyme involved in collagen metabolism, may be one of the targets for gentamicin-induced inhibition of collagen biosynthesis. We found that gentamicin-induced inhibition of prolidase activity (IC(50) approximately 100 microM) and collagen biosynthesis (IC(50) approximately 100 microM). At this concentration of gentamicin, DNA biosynthesis in human skin fibroblasts was inhibited only by about 30%. Melanin at 100 microg/ml produced about 25% inhibition of DNA synthesis and about 30% inhibition of prolidase activity, but it had no effect on collagen biosynthesis in cultured fibroblasts. However, the addition of melanin (100 microg/ml) to gentamicin-treated cells (100 microM) augmented the inhibitory action of gentamicin on collagen and DNA biosynthesis and partially reversed its inhibitory effect on prolidase activity. A melanin-induced augmentation of the inhibitory effects of gentamicin on collagen and DNA biosynthesis may explain the mechanism for the organ specificity of gentamicin-induced hearing loss in patients administered this drug. PMID:12098580

Wrze?niok, Dorota; Buszman, Ewa; Karna, Ewa; Nawrat, Piotr; Palka, Jerzy

2002-06-20

94

Cell chip-based monitoring of toxic effects of cosmetic compounds on skin fibroblast cells.  

Science.gov (United States)

The present study estimated the efficacy of electrochemical detection of imidazolidinyl urea-induced cell toxicity in skin human fibroblast cells (HFF cells). The gold nanopunct structures were fabricated through a nanoporous alumina mask, and the structural formations were confirmed via scanning electron microscopy. The HFF cells were allowed to attach to RGD (Arg-Gly-Asp) peptide nanopatterned surfaces, and electrochemical tools were applied to skin cells attached to the chip surface. The HFF cells evidenced inflammation responses to allergens such as imidazolidinyl urea. The cells were subsequently treated with different concentrations of imidazolidinyl urea for 24 h in culture, which induced a change in the cyclic voltammetry (CV) current peak. Treatment with imidazolidinyl urea induced a loss of cell viability and accelerated inflammation in a concentration-dependent manner. The expression level of inflammation-related proteins such as IL-1 beta were increased in imidazolidinyl urea-treated cells. The CV results demonstrated that imidazolidinyl urea significantly reduced the current peaks in a dose-dependent manner. The results showed that the current peak was reduced in accordance with the increases in imidazolidinyl urea-induced inflammation. In conclusion, the results of this study suggest that the electrochemical-based chip provides crucial information for improvements to a cell chip system for drug screening applications. PMID:22966535

An, Jeung Hee; Lee, Jin-Sung; Chun, Je-Ran; Oh, Byung-Keun; Kafi, M D Abdul; Choi, Jeong-Woo

2012-07-01

95

Relationship between the in vitro radiosensitivity of skin fibroblasts and the expression of subcutaneous fibrosis, telangiectasia, and skin erythema after radiotherapy  

International Nuclear Information System (INIS)

Objective: To investigate if the occurrence of subcutaneous fibrosis after radiotheraphy in an unselected group of breast cancer patients is related to cellular radiosensitivity of skin fibroblasts as measured in a clonogenic assay. Materials and methods: An in vitro colony-forming assay of normal fibroblast radiosensitivity was applied to primary skin biopsies from 31 breast cancer patients who received post-mastectomy radiotherapy with large doses per fraction (2.7-3.9 Gy) more than 10 years earlier. Three clinical normal-tissue endpoints were assessed. Two late endpoints, subcutaneous fibrosis and telangiectasia, were evaluated in three treatments fields by a single experienced clinician. In addition, skin erythema had been assessed at the end of treatment by members of the staff and junior staff. >From previous analyses of normal tissue response, individual clinical radiosensitivity could be assessed as 'excess risk' of each of the three reactions. This was defined as the difference between the actual observed response in the patient and the expected response estimated from individual treatment characteristics in a linear quadratic (LQ) mixture model and, for the two late endpoints, with correction for the follow-up time. This clinical radioresponsiveness was compared with the in vitro radiosensitivity of the skin fibroblasts. To this end, the fractions of colony-forming cells after graded single doses were fitted by an LQ survival curve using non-linear and linea survival curve using non-linear and linear regression from which the surviving fraction at 3.5 Gy (SF3.5) was estimated. Assessment at 3.5 Gy was chosen to reflect the fraction size during clinical radiotherapy. Results: A statistically significant variability of in vitro radiosensitivity between patients could be detected for both SF2 (P = 0.0095) and SF3.5 (P = 0.0008). A significant correlation was observed between SF3.5 and excess risk of fibrosis (rs -0.46, P = 0.009) while no association was found between fibroblast radiosensitivity and either the occurrence of severe skin telangiectasia or the acute endpoint skin erythema. Conclusion: These results suggest that variability in the occurrence of subcutaneous fibrosis, but not telangiectasia or erythema, after radiotherapy is partly accounted for by differences in cellular radiosensitivity of normal skin fibroblasts

96

The stimulatory effects of alpha1-adrenergic receptors on TGF-beta1, IGF-1 and hyaluronan production in human skin fibroblasts.  

Science.gov (United States)

Skin fibroblasts modulate tissue repair, wound healing and immunological responses. Adrenergic receptors (ARs) mediate important physiological functions, such as endocrine, metabolic and neuronal activity. In this study, the expression ?1A-ARs in human skin fibroblasts is examined and verified. Regulatory effects of ?1-agonist cirazoline on cell migration and the production of transforming growth factor ?1 (TGF-?1), insulin-like growth factor 1 (IGF-1), hyaluronan (HA), fibronectin and procollagen type I carboxy-terminal peptide (PIP) by human skin fibroblasts are assessed and validated. ?1A-AR mRNA and protein were found in human skin fibroblasts WS1. Exposure of cirazoline doubled skin fibroblast migration and the increase in cell migration was attenuated by ?1-antagonist prazosin. TGF-?1 mRNA and production were enhanced after exposure to cirazoline and IGF-1 production was also increased after treatment with cirazoline. Exposure to cirazoline also enhanced HA and PIP production. The increases in TGF-?1, IGF-1, HA and PIP production were partially abolished in fibroblasts transfected with ?1A-AR short interfering RNAs, indicating that ?1A-ARs are involved in the cirazoline-induced increases in TGF-?1, IGF-1, HA and PIP production. Thus, ?1A-ARs are stably expressed and stimulate cell migration and TGF-?1, IGF-1, HA and PIP production in human skin fibroblasts. Moreover, TGF-?1, IGF-1, HA and PIP production and the cell migration of human skin fibroblasts are possibly modulated by natural catecholamines produced by the endocrine system or sympathetic innervation, which could directly or indirectly participate in cytokine secretion, fibroblast migration and matrix production of wound healing in the skin. PMID:24844469

Liao, Ming-Huei; Liu, Shyh-Shyan; Peng, I-Chin; Tsai, Feng-Jen; Huang, Han Hsiang

2014-09-01

97

Thyroid hormone excess stimulates the synthesis of proteoglycan in human skin fibroblasts in culture  

International Nuclear Information System (INIS)

We previously demonstrated that proteoglycan accumulated in the affected skin of circumscribed pretibial myxedema of Graves' disease. As an underlying mechanism responsible for the accumulation, we sought to determine whether excess thyroid hormone was partially responsible for the increase in proteoglycan synthesis. Human skin fibroblasts were cultured in Ham's F-10 medium containing 1% Nutridoma with graded doses of T3(0.184 x 10-9 to 46 x 10-9 mol/l) and were labelled with [35S]sulphate and [3H]glucosamine. Proteoglycans were purified by Sephadex G-50, Q-Sepharose chromatography with NaCl-gradient and Sepharose CL-6B chromatography. 35S and 3H incorporated into dermatan sulphate proteoglycan and heparan sulphate proteoglycan and 3H incorporated into hyaluronan were measured. 35S and 3H incorporation into dermatan sulphate proteoglycan was minimum at a T3 concentration of 0.184 x 10-9 mol/l, and increased with increasing doses of T3 up to 46 x 10-9 mol/l. 35S and 3H incorporation into heparan sulphate proteoglycan also increased with increasing-doses of T3. 3H incorporation into hyaluranan was not influenced at all by T3. The increased incorporation of 35S into proteoglycan in high-T3 culture reflects the increased synthesis of proteoglycacts the increased synthesis of proteoglycan because 1. the extent of sulphation of disaccharides examined by thin-layer chromatography was not altered by T3; 2. the specific activity of [35S]sulphate was not influenced by T3, and 3. T3 did not decrease the degradation rate of cell-associated proteoglycan. (author)

98

The effect of hyaluronic acid on interleukin-1-induced deregulation of collagen metabolism in cultured human skin fibroblasts.  

Science.gov (United States)

Although hyaluronic acid (HA) has been used in the treatment of osteoarthritis for 30 years, the mechanism of its protective action on collagen metabolism disturbances in tissues during inflammation is not known. The present study was undertaken to evaluate the mechanism of Interleukin-1 (IL-1)-induced deregulation of collagen metabolism in cultured human skin fibroblast and the effect of HA on the process. In normal fibroblasts IL-1 strongly induced inhibition of collagen biosynthesis, while HA counteracted the process. The mechanism of this phenomenon was independent of prolidase activity, an enzyme that plays an important role in collagen biosynthesis at the post-translational level. Instead, IL-1 was found to inhibit the expression of insulin-like growth factor-I receptor (IGF-IR) and MAP kinases-ERK1 and ERK2, while HA was shown to counteract this process. Since insulin-like growth factor-I (IGF-I) is a most potent stimulator of collagen biosynthesis in fibroblasts the mechanism of IL-1-dependent inhibition of collagen biosynthesis may be related to inhibition of IGF-IR expression and signaling. The data suggest that hyaluronic acid protects collagen against IL-1-induced inhibition of biosynthesis of this protein in cultured human skin fibroblasts at the level of IGF-IR signaling. PMID:15749462

Nawrat, P; Surazy?ski, A; Karna, E; Pa?ka, J A

2005-05-01

99

YAG laser treatment causes rapid degeneration and regeneration of collagen fibres in pig skin and facilitates fibroblast growth.  

Science.gov (United States)

The non-ablative laser therapies have been speculated to cause microinjury in the dermal collagen fibres and increase collagen synthesis in the fibroblasts, leading to remodelling of the extracellular matrix. This study investigated the effects of neodymium YAG laser treatment on pig skin, especially focusing on its extracellular matrix molecules. The dorsal areas of a minipig were subjected to laser treatment, and samples were obtained by punch biopsies, and histological, immunohistochemical, and biochemical analyses were performed. The laser treatment caused degeneration of collagen fibres and fibrils, which were reconstituted within 24 hours, whereas there was no inflammation and no apparent damage on elastic fibres. Small blood vessels disappeared by the laser treatment, which re-appeared in 3 days. Biochemically, the amounts of collagen decreased up to day 3 after the treatment and then increased at day 7. When fibroblasts in dermal tissue at day 28 were counted, more fibroblasts in the treated tissue were observed than non-treated control. These results suggest that, although the laser treatment transiently degenerates collagen fibres and fibrils, it restores and increases them, mainly by an increase in dermal fibroblasts, assuring its minimal complication of skin. PMID:22998144

Kono, Ayuko; Oguri, Akiko; Yokoo, Kazuhisa; Watanabe, Hideto

2012-10-01

100

Skin Equivalent Tissue-Engineered Construct: Co-Cultured Fibroblasts/ Keratinocytes on 3D Matrices of Sericin Hope Cocoons  

OpenAIRE

The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from “Sericin Hope?...

Nayak, Sunita; Dey, Sancharika; Kundu, Subhas C.

2013-01-01

101

Photobiomodulation on the proliferation and collagen synthesis of normal human skin fibroblast cells  

Science.gov (United States)

Background and Objective: Cultured normal human skin fibroblast cells (HSFs) were once used to study the mechanism of the effects of low intensity He-Ne laser irradiation (LHNL) on wound healing. The proliferation and collagen synthesis of HFSs were modulated by LHNL in different papers, respectively, and both of them are studied in this paper. Study Design/Materials and Methods: The dosage was studied for the same radiation time 300s. The proliferation and collagen synthesis were measured by 3-[4,5-Dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay and the spectrophotometric method for the determination of hydroxyproline, respectively. Results: The dose zones were called dose 1, dose 2 and dose 3 from low dose on so that HSF proliferation was inhibited in dose 1 (16, 24 mJ/cm2), and promoted in dose 2 (298, 503, 597mJ/cm2), and the collagen synthesis was inhibited in dose 2 (401, 526 mJ/cm2), and promoted in dose 3 (714, 926, 1539, 1727mJ/cm2), which supports our biological model of photobiomodulation. It was found there is the linear relationship of the effect with dose with dose in each dose zone. Conclusions: The photobiomodulation on the proliferation and collagen synthesis of HSFs might be linearly dose-dependent in limited dosage with radiation time kept constant, which provides a foundation to discuss photobiomodulation on wound healing.

Cheng, Lei; Liu, Timon Cheng-Yi; Chi, Jin-Quan; Li, Yan; Jin, Hua

2006-01-01

102

Radiosensitivity of skin fibroblasts from atomic bomb survivors with and without breast cancer  

International Nuclear Information System (INIS)

Fibroblasts were established in vitro from skin biopsies obtained from 55 women and one man with or without breast cancer and with or without exposure to radiation from the atomic bomb explosion in Hiroshima. The radiosensitivity of these cells was evaluated by clonogenic assays after exposure to X rays or to fission neutrons from a 252Cf source. Data were fitted to a multitarget model, S/S0 = A[1-(1-ekD)N], for both X-ray and neutron dose-survival curves. A single-hit model, S/S0 = AekD, fits the neutron dose-survival responses as well. These was no difference in the means or variances of radiosensitivity between exposed and nonexposed groups, or between patients with or without breast cancer. Hence, although the sample is not large, it provides no support for the hypothesis that A-bomb radiation preferentially induces breast cancer in women whose cells in vitro are sensitive to cell killing by radiation. (author)

103

The expression of presenilin-1 mRNA in skin fibroblasts from Alzheimer's disease.  

Science.gov (United States)

The presenilin-1 (PS-1) gene was recently identified as one of the causative genes in the early onset of familial Alzheimer's disease (AD). Analysis of the PS-1 gene is thought to be useful in clarifying the pathogenesis of AD. However, there have been few reports about the expression of the PS-1 gene in AD. In this study, we analyzed the expression of PS-1 mRNA in cultured skin fibroblasts taken from living patients with AD by Northern blot analysis. The subjects consisted of 18 cases with AD and 10 cases of neurological patients without dementia (CTL). We found that the PS-1 mRNA levels in AD were significantly higher than those in CTL (p < 0.01). Moreover, we found that the PS-1 mRNA level increases in the early stages of AD and tends to decrease in the advanced stages. These findings suggest that high levels of PS-1 mRNA may play an important role in developing AD. PMID:9622002

Ikeda, K; Urakami, K; Isoe, K; Ohno, K; Nakashima, K

1998-01-01

104

Gene expression differences in skin fibroblasts in identical twins discordant for type 1 diabetes.  

Science.gov (United States)

Clinical studies suggest metabolic memory to hyperglycemia. We tested whether diabetes leads to persistent systematic in vitro gene expression alterations in patients with type 1 diabetes (T1D) compared with their monozygotic, nondiabetic twins. Microarray gene expression was determined in skin fibroblasts (SFs) of five twin pairs cultured in high glucose (HG) for ?6 weeks. The Exploratory Visual Analysis System tested group differences in gene expression levels within KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways. An overabundance of differentially expressed genes was found in eight pathways: arachidonic acid metabolism (P = 0.003849), transforming growth factor-? signaling (P = 0.009167), glutathione metabolism (P = 0.01281), glycosylphosphatidylinositol anchor (P = 0.01949), adherens junction (P = 0.03134), dorsal-ventral axis formation (P = 0.03695), proteasome (P = 0.04327), and complement and coagulation cascade (P = 0.04666). Several genes involved in epigenetic mechanisms were also differentially expressed. All differentially expressed pathways and all the epigenetically relevant differentially expressed genes have previously been related to HG in vitro or to diabetes and its complications in animal and human studies. However, this is the first in vitro study demonstrating diabetes-relevant gene expression differences between T1D-discordant identical twins. These SF gene expression differences, persistent despite the HG in vitro conditions, likely reflect "metabolic memory", and discordant identical twins thus represent an excellent model for studying diabetic epigenetic processes in humans. PMID:22315306

Caramori, M Luiza; Kim, Youngki; Moore, Jason H; Rich, Stephen S; Mychaleckyj, Josyf C; Kikyo, Nobuaki; Mauer, Michael

2012-03-01

105

Cytotoxic and mutagenic effects of high let charged particles on human skin fibroblasts  

International Nuclear Information System (INIS)

Cytotoxic and mutagenic effects of high LET charged particles were quantitatively measured using primary cultures of human skin fibroblasts. The span of LETs selected were from 25 keV/?m(330 MeV/u) to 920 keV/?m (600 MeV/u). Mutations were scored at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus using 6-thioguanine (6-TG) for selection. Exposure to these high LET charged particles resulted in exponential survival curves whereas mutant induction was fitted by a linear model. The Relative Biological Effect (RBE) for cell-killing ranged from 3.73 to 1.25, while that for mutant induction ranged from 5.74 to 0.48. Maximum RBE values were obtained at the LET of 150 keV/?m. The inactivation cross-section (?i) and the action cross-section for mutant induction (?m) ranged from 2.2 to 92.0 ?m2 and 0.09 to 5.56 x 10-3 ?m2 respectively, the maximum values were obtained by 56Fe with an LET of 200 keV/?m. The mutagenicity (?m/?i) ranged from 2.05 to 7.99 x 10-5 with the maximum value at 150 keV/?m. Furthermore, the results of multiplex polymerase chain reaction (PCR) of some of the mutants induced by charged particles indicate that higher LET beams are more likely to cause larger deletions in the hprt locus. (author)

106

Metabolism of cerebroside sulfate and subcellular distribution of its metabolites in cultured skin fibroblasts from controls, metachromatic leukodystrophy, and globoid cell leukodystrophy.  

OpenAIRE

With pulse-chase study of 1-[14C]stearic acid-labeled cerebroside sulfate (14C-CS) and subsequent subcellular fractionation by Percoll gradient, the metabolism of CS and translocation of its metabolites in human skin fibroblasts from controls, metachromatic leukodystrophy (MLD), and globoid cell leukodystrophy (GLD) were studied. In control skin fibroblasts, CS was transported to lysosome and metabolized there to galactosylceramide (GalCer) and ceramide (Cer) within 1 h. During the chase peri...

Inui, K.; Furukawa, M.; Okada, S.; Yabuuchi, H.

1988-01-01

107

Studies of the UVC-sensitivity of non-tumorigenic and tumorigenic human cell hybrids (HeLa x skin fibroblasts)  

International Nuclear Information System (INIS)

The UVC-sensitivities of a non-tumorigenic and a tumorigenic human cell hybrid (HeLa x skin fibroblasts) are compared and contrasted. The tumorigenic cells differ from the non-tumorigenic cells in that they have lost one copy each of chromosomes 11 and 14. For exponentially growing cultures, the tumorigenic cells are considerably more resistant than the non-tumorigenic cells. For confluent cultures, the differential in photosensitivity is much less. Flow cytometric studies of cell cycle distributions of both exponentially growing and confluent cultures of these cells indicate that the differences in photosensitivity cannot be explained by differences in cell cycle distribution. Studies of the kinetics of potentially lethal damage repair (PLDR) in confluent cultures of both cell lines indicate little or no recovery over the first 6h followed by an increase in survival over the next 12-24h. These data are consistent with previously published observations in human skin fibroblasts where the kinetics of PLDR reflected the kinetics of thymine dimer loss. The data are not consistent with 6-4 photoproducts being a potentially lethal lesion since such damage is rapidly repaired in human cells. (author)

108

Molecular alterations of tropoelastin and proteoglycans induced by tobacco smoke extracts and ultraviolet A in cultured skin fibroblasts  

International Nuclear Information System (INIS)

Functional integrity of normal skin is dependent on the balance between the biosynthesis and degradation of extracellular matrix, primarily composed of collagen, elastin and proteoglycans. In our previous studies, we found that tobacco smoke extracts decreased expressions of type I and III procollagen and induced matrix metalloproteinase-1 (MMP-1) and MMP-3 in the cultured skin fibroblasts. We here further investigated the effects of tobacco smoke extracts or ultraviolet A (UVA) treatments on the expression of tropoelastin (soluble elastin protein), and versican and decorin (proteoglycans) in cultured skin fibroblasts. The mRNA of tropoelastin increased by tobacco smoke extracts or UVA irradiation. Versican was markedly shown to decrease after these treatments by using western blotting and the mRNA of versican V0 also significantly decreased. UVA treatment did not show remarkable change in decorin protein, but resulted in marked decrease of decorin D1 mRNA. In contrast to UVA irradiation, the treatments of tobacco smoke extracts resulted in significant increase in decorin, while mRNA of decorin D1 decreased as compared to the control. MMP-7 increased after the treatment of tobacco smoke extracts or UVA. These results indicated that common molecular features might underlie the skin premature aging induced by tobacco smoke extracts and UVA, including abnormal regulation of extracellular matrix deposition through elevated MMPs, reduced collagen production, abnormal tropoeduced collagen production, abnormal tropoelastin accumulation, and altered proteoglycans. (author)

109

In vitro study of the impact of mechanical tension on the dermal fibroblast phenotype in the context of skin wound healing.  

Science.gov (United States)

Skin wound healing is finely regulated by both matrix synthesis and degradation which are governed by dermal fibroblast activity. Actually, fibroblasts synthesize numerous extracellular matrix proteins (i.e., collagens), remodeling enzymes and their inhibitors. Moreover, they differentiate into myofibroblasts and are able to develop endogenous forces at the wound site. Such forces are crucial during skin wound healing and have been widely investigated. However, few studies have focused on the effect of exogenous mechanical tension on the dermal fibroblast phenotype, which is the objective of the present paper. To this end, an exogenous, defined, cyclic and uniaxial mechanical strain was applied to fibroblasts cultured as scratch-wounded monolayers. Results showed that fibroblasts' response was characterized by both an increase in procollagen type-I and TIMP-1 synthesis, and a decrease in MMP-1 synthesis. The monitoring of scratch-wounded monolayers did not show any decrease in kinetics of the filling up when mechanical tension was applied. Additional results obtained with proliferating fibroblasts and confluent monolayer indicated that mechanical tension-induced response of fibroblasts depends on their culture conditions. In conclusion, mechanical tension leads to the differentiation of dermal fibroblasts and may increase their wound-healing capacities. So, the exogenous uniaxial and cyclic mechanical tension reported in the present study may be considered in order to improve skin wound healing. PMID:25267573

Rolin, Gwenae L; Binda, Delphine; Tissot, Marion; Viennet, Céline; Saas, Philippe; Muret, Patrice; Humbert, Philippe

2014-11-01

110

Endogenous glutathione protects human skin fibroblasts against the cytotoxic action of UVB, UVA and near-visible radiations  

International Nuclear Information System (INIS)

Both the UVB (290-320 nm) and UVA (320-380 nm) regions of sunlight damage human skin cells but, particularly at the longer wavelengths, information is scant concerning the mechanism(s) of damage induction and the roles of cellular defense mechanisms. Following extensive glutathione depletion of cultured human skin fibroblasts, the cells become strongly sensitized to the cytotoxic action of near-visible (405 nm), UVA (334 nm, 365 nm) and UVB (313 nm) but not UVC (254 nm) radiations. In the critical UVB region, the magnitude of the protection afforded by endogenous glutathione approaches that of the protection provided by excision repair. The results suggest that a significant fraction of even UVB damage can be mediated by free radical attack and that a major role of glutathione in human skin cells is to protect them from the cytotoxic action of sunlight. (author)

111

Influence of beam shape on in-vitro cellular transformations in human skin fibroblasts  

Science.gov (United States)

A variety of strategies have been utilised for prevention and treatment of chronic wounds such as leg ulcers, diabetic foot ulcers and pressure sores1. Low Level Laser Therapy (LLLT) has been reported to be an invaluable tool in the enhancement of wound healing through stimulating cell proliferation, accelerating collagen synthesis and increasing ATP synthesis in mitochondria to name but a few2. This study focused on an in-vitro analysis of the cellular responses induced by treatment with three different laser beam profiles namely, the Gaussian (G), Super Gaussian (SG) and Truncated Gaussian (TG), on normal wounded irradiated (WI) and wounded non-irradiated (WNI) human skin fibroblast cells (WS1), to test their influence in wound healing at 632.8 nm using a helium neon (HeNe) laser. For each beam profile, measurements were made using average energy densities over the sample ranging from 0.2 to 1 J, with single exposures on normal wounded cells. The cells were subjected to different post irradiation incubation periods, ranging from 0 to 24 hours to evaluate the duration (time) dependent effects resulting from laser irradiation. The promoted cellular alterations were measured by increase in cell viability, cell proliferation and cytotoxicity. The results obtained showed that treatment with the G compared to the SG and TG beams resulted in a marked increase in cell viability and proliferation. The data also showed that when cells undergo laser irradiation some cellular processes are driven by the peak energy density rather than the energy of the laser beam. We show that there exist threshold values for damage, and suggest optimal operating regimes for laser based wound healing.

Mthunzi, Patience; Forbes, Andrew; Hawkins, Denise; Abrahamse, Heidi; Karsten, Aletta E.

2005-08-01

112

Elongation of 20-carbon polyunsaturated fatty acids by human skin fibroblasts  

International Nuclear Information System (INIS)

Human skin fibroblasts readily incorporate exogenous arachidonate (20:4(n-6)) and eicosapentaenoate (20:5(n-3)) into cellular phospholipids and triacylglycerol. The extent of incorporation of 1.25 ?M [14C]20:4(n-6) and [14C]20:5(n-3) from culture medium with delipidized serum protein is similar, 20% in 1 hr increasing to 60-70% within 8 hr. Elongation of incorporated [14C]20:5(n-3) to [14C]22:5(n-3) is extensive, 40% by 8 hr and 85% by 48 hr. Elongation of [14C]20:4(n-6) to [14C]22:4(n-6) is 1$C]20:5(n-3) and plateaus at ?20% of incorporated 14C-fatty acid. Although incorporated, exogenous 22:4(n-6) is not an effective inhibitor of the elongation of [14C]arachidonate; however, exogenous 20:5(n-3) is inhibitory, with an ID50 of 5 ?M. With exogenous concentrations of [14C]arachidonate from 0.4 - 10 ?M, the percentage incorporated in 24 hr remains relatively constant. By contrast, the extent of elongation of incorporated [14C]arachidonate increases from 12% at 0.4 ?M to 43% at 10 ?M. Under these conditions, elongation of incorporated [14C]20:5(n-3) is ?75%. Thus, in these cells, selectivity of the elongation system results in differential metabolism of 20-carbon n-6 and n-3 fatty acids. Furthermore, arachidonate appears to act as a positive modulator of its own elongation

113

Human bone marrow stromal cells and skin fibroblasts inhibit natural killer cell proliferation and cytotoxic activity.  

Science.gov (United States)

Mesenchymal stromal cells (MSCs) are potent immunomodulators that have successfully been used to circumvent various types of inflammations, including steroid-resistant graft-versus-host disease. Although initially believed to be restricted to multipotent MSCs, this immunoregulatory function is shared with differentiated cells from the mesenchymal lineage such as skin fibroblasts (SFs). Mesenchymal cell-induced immunoregulation is so potent that it may allow the reactivation of dormant malignancies, a fact that would preclude using such cells as therapeutic agents. Because NK cells are pivotal effectors controlling tumor cell containment we investigated the effect of allogenic MSCs and SFs on NK cell function in vitro. When NK cells were incubated with IL-15 and MSCs or SFs for 6 days, their proliferation and cytotoxic activity were significantly decreased compared to NK cells cultured with IL-15 alone or with human venous endothelial cells. Cytotoxic activity inhibition reached 86% when assayed on MHC-I(+) allogenic primary hematopoietic blasts, and was associated with a significant decrease in cytolytic granule exocytosis and in perforin release. Stromal cell-mediated inhibition was effective only if cell-cell proximity was long lasting: when NK cells were activated with IL-15 in the absence of MSCs and assayed for cytotoxicity in their presence no inhibition occurred. MSC inhibition was ultimately mediated by a soluble factor generated upon incubation with NK cells activated by IL-15 or IL-2. The indoleamine 2,3 dioxygenase was activated in MSCs and SFs because L-kynurenine was detected in inhibitory supernatants, but its blockade did not restore NK cell functions. The profound inhibition of cytotoxic activity directed against allogenic hematopoietic blasts exerted by MSCs and SFs on NK cells may be a concern. Should this occur in vivo it may induce the inability of NK cells to control residual or dormant malignant diseases after infusion of therapeutic MSCs. PMID:21054933

Pradier, Amandine; Passweg, Jakob; Villard, Jean; Kindler, Vincent

2011-01-01

114

Photosensitization of skin fibroblasts and HeLa cells by three chlorin derivatives: Role of chemical structure and delivery vehicle.  

Science.gov (United States)

The chemical nature of the sensitizer and its selective uptake by malignant cells are decisive to choose an appropriate biocompatible carrier, able to preserve the photosensitizing characteristics of the dye. In this paper we demonstrate the photodynamic properties of three chlorins, derived from chlorophyll a, and the usefulness of liposomal carriers to design pharmaceutical formulations. The chlorins have been quantitatively incorporated into stable liposomes obtained from a mixture of L-alpha-palmitoyloleoylphosphatidylcholine and L-alpha-dioleoylphosphatidylserine in a 13.5:1.5 molar ratio (POPC/OOPS-liposomes). The chlorin uptake by skin fibroblasts increases steadily, reaching in all cases a plateau level dependent on both the chlorin structure and the vehicle employed. The photophysical properties of the three chlorins in THF are nearly identical and fulfill the requirements for a PDT photosensitizer. Incorporation of chlorins into liposomes induces important changes in their photophysics, but does not impair their cellular uptake or their cell photosensitization ability. In fact we observe in the cells the same photophysical behavior as in THF solution. Specifically, we demonstrate, by recording the near-IR phosphorescence of 1O2, that the chlorins are able to photosensitize the production of 1O2 in the cell membrane. The cell-photosensitization efficiency depended on the chlorin and cell line nature, the carrier, and the length of pre-incubation and post-irradiation periods. The high photodynamic activity of chlorin-loaded liposomes and the possibility to design liposomal carriers to achieve a specific target site favors this approach to obtain an eventual pharmaceutical formulation. PMID:16740249

Postigo, Fernando; Sagristá, M Luisa; De Madariaga, M Africa; Nonell, Santi; Mora, Margarita

2006-05-01

115

Carnosic acid, a phenolic diterpene from rosemary, prevents UV-induced expression of matrix metalloproteinases in human skin fibroblasts and keratinocytes.  

Science.gov (United States)

Exposure of the skin to ultraviolet (UV) irradiation induces photoageing through the up-regulation of matrix metalloproteinases (MMPs) and subsequent breakdown of extracellular matrices. Reactive oxygen species (ROS) and epidermal growth factor receptor (EGFR) activation play central roles in UV-induced MMP expression through initiating extracellular signal-regulated kinase (ERK)-mediated AP-1 signalling. We aimed to explore the effects of carnosic acid (CA), a phenolic diterpene from rosemary, on UV-induced MMP expression in human skin cells. Molecular mechanism underlying the effects of CA was also examined in the aspect of MMP expression, ERK/AP-1 pathway, ROS generation and EGFR activation. Human dermal fibroblast cell line (Hs68), primary normal human dermal fibroblasts (HDFs) and normal human epidermal keratinocytes (HEKs) were employed, and antiphotoageing effects of CA were assessed by Western blotting, quantitative real-time PCR and enzyme assays. CA significantly inhibited UVA- and UVB-induced expression of MMP-1, MMP-3 and MMP-9 in a concentration-dependent manner in Hs68 cells. UVB-induced ERK activation and the formation of transcription factor, AP-1, were significantly suppressed by CA. Among the upstream events of MMP expression, UVB-induced ROS generation was attenuated by CA, while EGFR activation was not affected. Confirming the antiphotoageing effects of CA through the suppression of UV-induced ROS generation, UVB-enhanced GADD45 expression, a marker for oxidative DNA damages was significantly reduced by CA. Inhibitory effects of CA on UVB-induced MMP expression could be also seen in HDFs and HEKs. Collectively, our study demonstrates that CA inhibits the UV-enhanced MMPs in human skin cells through the inhibition of ROS and the suppression of ERK/AP-1 activation. PMID:23614740

Park, Miyoung; Han, Jiwon; Lee, Chang Seok; Soo, Baek Heung; Lim, Kyung-Min; Ha, Hunjoo

2013-05-01

116

Acute leukemia after radiotherapy in a patient with Turcot's syndrome. Impaired colony formation in skin fibroblast cultures after irradiation  

International Nuclear Information System (INIS)

Colonic polyposis and carcinoma developed in a woman with Turcot's syndrome at the age of 31 years; astrocytoma developed when she was 37. Her brother and sister had died of astrocytoma at the ages of 18 and 33 years, respectively. Progressive neutropenia developed in the patient three months after radiotherapy for her brain tumor and acute myelomonocytic leukemia 19 months after treatment. Three laboratories independently evaluated cultures of her skin fibroblasts for in vitro sensitivity to cell killing (loss of colony-forming ability) by x-rays. Survival assays consistently revealed slight but significant radiosensitivity in an early-passage (six to 10 doublings) fibroblast subculture. A later subculture (21 to 29 doublings) showed no abnormality, a possible effect of selective in vitro loss of radiosensitive cells

117

Radiation induced DNA damage and damage repair in human tumor and fibroblast cell lines assessed by histone H2AX phosphorylation  

International Nuclear Information System (INIS)

Purpose: To analyze the radiation-induced levels of ?H2AX and its decay kinetics in 10 human cell lines covering a wide range of cellular radiosensitivity (SF2, 0.06-0.63). Methods and Materials: Five tumor cell lines included Colo-800 melanoma, two glioblastoma (MO59J and MO59K), fibrosarcoma HT 1080, and breast carcinoma MCF7. Five primary skin fibroblasts lines included two normal strains, an ataxia telangiectasia strain, and two fibroblast strains from breast cancer patients with an adverse early skin reaction to radiotherapy. Cellular radiosensitivity was assessed by colony-forming test. Deoxyribonucleic acid damage and repair were analyzed according to nuclear ?H2AX foci intensity, with digital image analysis. Results: The cell lines tested showed a wide degree of variation in the background intensity of immunostained nuclear histone ?H2AX, which was higher for the tumor cell lines compared with the fibroblast strains. It was not possible to predict clonogenic cell survival (SF2) for the 10 cell lines studied from the radiation-induced ?H2AX intensity. In addition, the slopes of the dose-response (0-4 Gy) curves, the rates of ?H2AX disappearance, and its residual expression (?18 h after irradiation) did not correlate with SF2 values. Conclusions: The results from 10 cell lines showed that measurements of immunofluorescence intensity by digital image analysis of phosphorylated histone H2AX as a surrogate marker of DNA double-strand breaks did not allow relble-strand breaks did not allow reliable ranking of cell strains according to their clonogenic survival after irradiation

118

Epilobium angustifolium extract demonstrates multiple effects on dermal fibroblasts in vitro and skin photo-protection in vivo.  

Science.gov (United States)

Stress-induced fibroblast senescence is thought to contribute to skin aging. Ultraviolet light (UV) radiation is the most potent environmental risk factor in these processes. An Epilobium angustifolium (EA) extract was evaluated for its capacity to reverse the senescent response of normal human dermal fibroblasts (NHDF) in vitro and to exhibit skin photo-protection in vivo. The HPLC-UV-MS analysis of the EA preparation identified three major polyphenol groups: tannins (oenothein B), phenolic acids (gallic and chlorogenic acids) and flavonoids. EA extract increased the cell viability of senescent NHDF induced by serum deprivation. It diminished connective tissue growth factor and fibronectin gene expressions in senescent NHDF. Down-regulation of the UV-induced release of both matrix metalloproteinase-1 and -3 and the tissue inhibitor of matrix metalloproteinases-1 and -2, and also down-regulation of the gene expression of hyaluronidase 2 were observed in repeatedly UV-irradiated NHDF after EA extract treatment. Interestingly, EA extract diminished the down-regulation of sirtuin 1 dampened by UV-irradiation. The application of EA extract using a sub-irritating dose protected skin against UV-induced erythema formation in vivo. In summary, EA extract diminished stress-induced effects on NHDF, particularly on connective tissue growth factor, fibronectin and matrix metalloproteinases. These results collectively suggest that EA extract may possess anti-aging properties and that the EA polyphenols might account for these benefits. PMID:23817638

Ruszová, Ema; Cheel, José; Pávek, Stanislav; Moravcová, Martina; Hermannová, Martina; Mat?jková, Ilona; Spilková, Ji?ina; Velebný, Vladimír; Kubala, Lukáš

2013-09-01

119

Fabrication of a nanofibrous scaffold with improved bioactivity for culture of human dermal fibroblasts for skin regeneration  

Energy Technology Data Exchange (ETDEWEB)

Engineering dermal substitutes with electrospun nanofibres have lately been of prime importance for skin tissue regeneration. Simple electrospinning technology served to produce nanofibrous scaffolds morphologically and structurally similar to the extracellular matrix of native tissues. The nanofibrous scaffolds of poly(l-lactic acid)-co-poly({epsilon}-caprolactone) (PLACL) and PLACL/gelatin complexes were fabricated by the electrospinning process. These nanofibres were characterized for fibre morphology, membrane porosity, wettability and chemical properties by FTIR analysis to culture human foreskin fibroblasts for skin tissue engineering. The nanofibre diameter was obtained between 282 and 761 nm for PLACL and PLACL/gelatin scaffolds; expressions of amino and carboxyl groups and porosity up to 87% were obtained for these fibres, while they also exhibited improved hydrophilic properties after plasma treatment. The results showed that fibroblasts proliferation, morphology, CMFDA dye expression and secretion of collagen were significantly increased in plasma-treated PLACL/gelatin scaffolds compared to PLACL nanofibrous scaffolds. The obtained results prove that the plasma-treated PLACL/gelatin nanofibrous scaffold is a potential biocomposite material for skin tissue regeneration.

Chandrasekaran, Arun Richard; Venugopal, J; Sundarrajan, S; Ramakrishna, S, E-mail: nnijrv@nus.edu.s [Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore (Singapore)

2011-02-15

120

Fabrication of a nanofibrous scaffold with improved bioactivity for culture of human dermal fibroblasts for skin regeneration  

International Nuclear Information System (INIS)

Engineering dermal substitutes with electrospun nanofibres have lately been of prime importance for skin tissue regeneration. Simple electrospinning technology served to produce nanofibrous scaffolds morphologically and structurally similar to the extracellular matrix of native tissues. The nanofibrous scaffolds of poly(l-lactic acid)-co-poly(?-caprolactone) (PLACL) and PLACL/gelatin complexes were fabricated by the electrospinning process. These nanofibres were characterized for fibre morphology, membrane porosity, wettability and chemical properties by FTIR analysis to culture human foreskin fibroblasts for skin tissue engineering. The nanofibre diameter was obtained between 282 and 761 nm for PLACL and PLACL/gelatin scaffolds; expressions of amino and carboxyl groups and porosity up to 87% were obtained for these fibres, while they also exhibited improved hydrophilic properties after plasma treatment. The results showed that fibroblasts proliferation, morphology, CMFDA dye expression and secretion of collagen were significantly increased in plasma-treated PLACL/gelatin scaffolds compared to PLACL nanofibrous scaffolds. The obtained results prove that the plasma-treated PLACL/gelatin nanofibrous scaffold is a potential biocomposite material for skin tissue regeneration.

121

Effect of wavelength and fluence on morphology, cellular and genetic integrity of diabetic wounded human skin fibroblasts  

Science.gov (United States)

An alternative treatment modality for diabetic wound healing includes low level laser therapy (LLLT). Biostimulation of such wounds may be of benefit to patients by reducing healing time. Structural, cellular and genetic events in diabetic wounded human skin fibroblasts (WS1) were evaluated after exposing cells in culture to a Helium-Neon (632.8nm), a Diode laser (830nm) and a Nd:YAG (Neodynium:Yttrium-Allumina-Gallium) laser (1064nm) at either 5J/cm2 or 16J/cm2. Cells were exposed twice a week and left 24 hours post-irradiation prior to measuring effects. Structural changes were evaluated by assessing colony formation, haptotaxis and chemotaxis. Cellular changes were evaluated using cell viability, (adenosine-triphosphate, ATP production), and proliferation, (alkaline phosphatase, ALP and basic fibroblast growth factor, bFGF expression), while the Comet assay evaluated DNA damage and cytotoxicity was determined assessing membrane permeability for lactate dehydrogenase (LDH). Caspase 3/7 activity was used as an estimate of apoptosis as a result of irradiation. The irradiated diabetic wounded cells showed structural, cellular as well as molecular resilience comparable to that of unwounded normal skin fibroblast cells. With regards to fluence, 5J/cm2 elicit positive cellular and structural responses while 16J/cm2 increases cellular and genetic damage and cellular morphology is altered. Different wavelengths of LLLT influences the beneficial outcomes of diabetic wounded cells and although all three wavelengths elicit cellular effects, the penetration depth of 830nm plays a significant role in the healing of diabetic wounded human fibroblast cells. Results from this study validate the contribution of LLLT to wound healing and elucidate the biochemical effects at a cellular level while highlighting the role of different dosages and wavelengths in LLLT.

Abrahamse, H.; Hawkins, D.; Houreld, N.

2006-02-01

122

Contact guidance of chick embryo neurons on single scratches in glass and on underlying aligned human skin fibroblasts.  

Science.gov (United States)

The influence of substratum topography on the morphology and orientation of neurites of chick embryo neurons was studied. Two series of experiments are reported. One concerned the behaviour of growth cones when the axons become contact-guided by the surface texture. The second studied contact guidance of neurites extending on a compact layer of fixed aligned human skin fibroblasts (HSF). It was observed that when the growth cones of sensory neurons isolated from dorsal root ganglions encountered a single scratch in a glass surface (0.1-2 microm in depth and diameter) they turned and continued movement following the axis of the scratch. These neurons became contact-guided as a result of the sequence of events. The growth cone filopodia recognized the irregularity in the substratum surface, whereas the growth cone lamella stabilized contact with the scratch and moved forward along the scratch axis. Scanning electron microscope revealed that the single scratches 150 nm in width and ca. 100 nm deep growth cone filopodia less than 200 nm in diameter could detect and react by turning into them. These filopodia extensions followed the edge of scratches. However, phase contrast and Nomarski's differential interference contrast appeared insufficient for analysis of primary contact guidance of fine growth cone filopodia which themselves are often less than 200 nm. In neuron cultures on fixed aligned HSF, the neuron aggregates assumed spindle-like shapes, and sparsely seeded individual neurons extended axons along the long axes of the fibroblasts. The axons extended significantly further on the fixed underlying fibroblasts than on collagen-covered glass. In crowded cultures of neurons, the cells extended neurites ignoring both the surface anisotropy (the scratches) and the orientation of the aligned fibroblasts. Immunofluorescence staining of neurons with antibodies against neurofilaments made it possible to analyse their shape and orientation on the fibroblasts. Computer-assisted image analysis permitted the observed alignment of the neurites to be characterized quantitatively. PMID:10561119

S?epie?, E; Stanisz, J; Korohoda, W

1999-01-01

123

Effect of bradykinin on prostaglandin production by human skin fibroblasts in culture  

International Nuclear Information System (INIS)

In studying the effect of bradykinin on prostaglandin formation in fibroblasts, two general types of assays have been employed. Radioimmunoassays were utilized to determine specific prostaglandins, PGI2 and 6-keto-PGF, using tritium as radiolabel. A second procedure involved initial incubation of the fibroblasts with [14C]arachidonate and identification and quantification of the metabolites by chromatographic methods. After radiolabelling of fibroblasts with [14C]arachidonate, bradykinin was found to release radiolabel from membrane lipids approximately 2-fold. In the presence of bradykinin, there was a close coupling of phospholipase S2 activity, arachidonate mobilization, and synthesis of a specific prostaglandin, PGI2

124

Nuclear uptake of 1,25-dihydroxy[3H]cholecalciferol in dispersed fibroblasts cultured from normal human skin.  

OpenAIRE

Because of the relative inaccessibility of known calciferol target tissues (i.e., intestine and bone), we examined fibroblasts derived from normal human skin and grown in tissue culture as a means of evaluating the interaction of 1,25-dihydroxycholecalciferol [1,25(OH)2D3] and its effector system. When dispersed, intact cells were used, nuclear uptake of 1,25-dihydroxy[23,24(n)3-H]cholecalciferol [1,25(OH)2[3H]D3) was temperature-dependent, optimal at 45 min at 37 degrees C, and saturable. In...

Eil, C.; Marx, S. J.

1981-01-01

125

Altered transcriptome signature of phenotypically normal skin fibroblasts heterozygous for CDKN2A in familial melanoma: relevance to early intervention  

OpenAIRE

Familial melanoma (FM) is a dominantly heritable cancer that is associated with mutations in the tumor suppressor CDKN2A/p16. In FM, a single inherited “hit” occurs in every somatic cell, enabling interrogation of cultured normal skin fibroblasts (SFs) from FM gene carriers as surrogates for the cell of tumor origin, namely the melanocyte. We compared the gene expression profile of SFs from FM individuals with two distinct CDKN2A/p16 mutations (V126D-p16 and R87P-p16) with the gene expres...

Fan, Meiyun; Pfeffer, Susan R.; Lynch, Henry T.; Cassidy, Pamela; Leachman, Sancy; Pfeffer, Lawrence M.; Kopelovich, Levy

2013-01-01

126

DNA damage in wounded, hypoxic and acidotic human skin fibroblast cell cultures after low laser irradiation  

Science.gov (United States)

Phototherapy has become more popular and widely used in the treatment of a variety of medical conditions. To ensure sound results as evidence of its effectiveness, well designed experiments must be conducted when determining the effect of phototherapy. Cell culture models such as hypoxic, acidotic and wounded cell cultures simulating different disease conditions including ischemic heart disease, diabetes and wound healing were used to determine the effect of laser irradiation on the genetic integrity of the cell. Even though phototherapy has been found to be beneficial in a wide spectrum of conditions, it has been shown to induce DNA damage. However, this damage appears to be repairable. The risk lies in the fact that phototherapy may help the medical condition initially but damage DNA at the same time leaving undetected damage that may result in late onset, more severe, induced medical conditions including cancer. Human skin fibroblasts were cultured and used to induce a wound (by the central scratch model), hypoxic (by incubation in an anaerobic jar, 95% N2 and 5% O2) and acidotic (reducing the pH of the media to 6.7) conditions. Different models were irradiated using a Helium-Neon (632.8 nm) laser with a power density of 2.07 mW/cm2 and a fluence of 5 J/cm2 or 16 J/cm2. The effect of the irradiation was determined using the Comet assay 1 and 24 h after irradiation. In addition, the Comet assay was performed with the addition of formamidopyrimidine glycosylase (FPG) obviating strand brakes in oxidized bases at a high fluence of 16 J/cm2. A significant increase in DNA damage was seen in all three injured models at both 1 and 24 h post-irradiation when compared to the normal un-injured cells. However, when compared to non-irradiated controls the acidotic model showed a significant decrease in DNA damage 24 h after irradiation indicating the possible induction of cellular DNA repair mechanisms. When wounded cells were irradiated with higher fluences of 16 J/cm2, there was a significant increase in DNA damage in irradiated cells with and without the addition of FPG. These results are indicative of the importance of both cell injury model as well as fluence when assessing the effect of phototherapy on DNA integrity.

Hawkins Evans, D.; Mbene, A.; Zungu, I.; Houreld, N.; Abrahamse, H.

2009-02-01

127

Cationic star-shaped polymer as an siRNA carrier for reducing MMP-9 expression in skin fibroblast cells and promoting wound healing in diabetic rats  

Science.gov (United States)

Background Excessive expression of matrix metalloproteinase-9 (MMP-9) is deleterious to the cutaneous wound-healing process in the context of diabetes. The aim of the present study was to explore whether a cationic star-shaped polymer consisting of ?-cyclodextrin (?-CD) core and poly(amidoamine) dendron arms (?-CD-[D3]7) could be used as the gene carrier of small interfering RNA (siRNA) to reduce MMP-9 expression for enhanced diabetic wound healing. Methods The cytotoxicity of ?-CD-(D3)7 was investigated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MMT) method in the rat CRL1213 skin fibroblast cell line. The transfection efficiency of ?-CD-(D3)7/MMP-9-small interfering RNA (siRNA) complexes was determined by confocal microscopy and flow cytometry. Quantitative real time (RT) polymerase chain reaction was performed to measure the gene expression of MMP-9 after the transfection by ?-CD-(D3)7/MMP-9-siRNA complexes. The ?-CD-(D3)7/MMP-9-siRNA complexes were injected on the wounds of streptozocin-induced diabetic rats. Wound closure was measured on days 4 and 7 post-wounding. Results ?-CD-(D3)7 exhibited low cytotoxicity in fibroblast cells, and easily formed the complexes with MMP-9-siRNA. The ?-CD-(D3)7/MMP-9-siRNA complexes were readily taken up by fibroblast cells, resulting in the downregulation of MMP-9 gene expression (P<0.01). Animal experiments revealed that the treatment by ?-CD-(D3)7/MMP-9-siRNA complexes enhanced wound closure in diabetic rats on day 7 post-wounding (P<0.05). Conclusion ?-CD-(D3)7 may be used as an efficient carrier for the delivery of MMP-9-siRNA to reduce MMP-9 expression in skin fibroblast cells and promote wound healing in diabetic rats. PMID:25075185

Li, Na; Luo, Heng-Cong; Yang, Chuan; Deng, Jun-Jie; Ren, Meng; Xie, Xiao-Ying; Lin, Diao-Zhu; Yan, Li; Zhang, Li-Ming

2014-01-01

128

Human lysosomes can be purified from diploid skin fibroblasts by free-flow electrophoresis.  

OpenAIRE

Human diploid fibroblasts underwent lysis when resuspended in isotonic sucrose. After preparation of a granular fraction by differential centrifugation, lysosomes were almost completely purified by free-flow electrophoresis. Electron micrographs of the final fraction of fibroblast lysosomes (about 25-fold purified) showed mostly secondary lysosomes having an appearance identical to those in intact cells. There was no indication that the purification steps selectd for any lysosomal subpopulati...

Harms, E.; Kern, H.; Schneider, J. A.

1980-01-01

129

Senescent Fibroblasts Enhance Early Skin Carcinogenic Events via a Paracrine MMP-PAR-1 Axis  

OpenAIRE

The incidence of carcinoma increases greatly with aging, but the cellular and molecular mechanisms underlying this correlation are only partly known. It is established that senescent fibroblasts promote the malignant progression of already-transformed cells through secretion of inflammatory mediators. We investigated here whether the senescent fibroblast secretome might have an impact on the very first stages of carcinogenesis. We chose the cultured normal primary human epidermal keratinocyte...

Malaquin, Nicolas; Vercamer, Chantal; Bouali, Fatima; Martien, Se?bastien; Deruy, Emeric; Wernert, Nicolas; Chwastyniak, Maggy; Pinet, Florence; Abbadie, Corinne; Pourtier, Albin

2013-01-01

130

Effects of sulfate deprivation on the production of chondroitin/dermatan sulfate by cultures of skin fibroblasts from normal and diabetic individuals  

International Nuclear Information System (INIS)

Human skin fibroblast monolayer cultures from two normal men, three Type I diabetic men, and one Type I diabetic woman were incubated with [3H]glucosamine in the presence of diminished concentrations of sulfate. Although total synthesis of [3H]chondroitin/dermatan glycosaminoglycans varied somewhat between cell lines, glycosaminoglycan production was not affected within any line when sulfate levels were decreased from 0.3 mM to 0.06 mM to 0.01 mM to 0 added sulfate. Lowering of sulfate concentrations resulted in diminished sulfation of chondroitin/dermatan in a progressive manner, so that overall sulfation dropped to as low as 19% for one of the lines. Sulfation of chondroitin to form chondroitin 4-sulfate and chondroitin 6-sulfate was progressively and equally affected by decreasing the sulfate concentration in the culture medium. However, sulfation to form dermatan sulfate was preserved to a greater degree, so that the relative proportion of dermatan sulfate to chondroitin sulfate increased. Essentially all the nonsulfated residues were susceptible to chondroitin AC lyase, indicating that little epimerization of glucuronic acid residues to iduronic acid had occurred in the absence of sulfation. These results confirm the previously described dependency of glucuronic/iduronic epimerization on sulfation, and indicate that sulfation of the iduronic acid-containing disaccharide residues of dermatan can take place with sulfate concentrations lower than those needed for 6-centrations lower than those needed for 6-sulfation and 4-sulfation of the glucuronic acid-containing disaccharide residues of chondroitin. There were considerable differences among the six fibroblast lines in susceptibility to low sulfate medium and in the proportion of chondroitin 6-sulfate, chondroitin 4-sulfate, and dermatan sulfate. However, there was no pattern of differences between normals and diabetics

131

Effects of sulfate deprivation on the production of chondroitin/dermatan sulfate by cultures of skin fibroblasts from normal and diabetic individuals  

Energy Technology Data Exchange (ETDEWEB)

Human skin fibroblast monolayer cultures from two normal men, three Type I diabetic men, and one Type I diabetic woman were incubated with (3H)glucosamine in the presence of diminished concentrations of sulfate. Although total synthesis of (3H)chondroitin/dermatan glycosaminoglycans varied somewhat between cell lines, glycosaminoglycan production was not affected within any line when sulfate levels were decreased from 0.3 mM to 0.06 mM to 0.01 mM to 0 added sulfate. Lowering of sulfate concentrations resulted in diminished sulfation of chondroitin/dermatan in a progressive manner, so that overall sulfation dropped to as low as 19% for one of the lines. Sulfation of chondroitin to form chondroitin 4-sulfate and chondroitin 6-sulfate was progressively and equally affected by decreasing the sulfate concentration in the culture medium. However, sulfation to form dermatan sulfate was preserved to a greater degree, so that the relative proportion of dermatan sulfate to chondroitin sulfate increased. Essentially all the nonsulfated residues were susceptible to chondroitin AC lyase, indicating that little epimerization of glucuronic acid residues to iduronic acid had occurred in the absence of sulfation. These results confirm the previously described dependency of glucuronic/iduronic epimerization on sulfation, and indicate that sulfation of the iduronic acid-containing disaccharide residues of dermatan can take place with sulfate concentrations lower than those needed for 6-sulfation and 4-sulfation of the glucuronic acid-containing disaccharide residues of chondroitin. There were considerable differences among the six fibroblast lines in susceptibility to low sulfate medium and in the proportion of chondroitin 6-sulfate, chondroitin 4-sulfate, and dermatan sulfate. However, there was no pattern of differences between normals and diabetics.

Silbert, C.K.; Humphries, D.E.; Palmer, M.E.; Silbert, J.E. (Veterans Administration Outpatient Clinic, Boston, MA (USA))

1991-02-15

132

Diagnosis of Metachromatic Leukodystrophy, Krabbe Disease, and Farber Disease after Uptake of Fatty Acid-labeled Cerebroside Sulfate into Cultured Skin Fibroblasts  

OpenAIRE

[14C]Stearic acid-labeled cerebroside sulfate (CS) was presented to cultured skin fibroblasts in the media. After endocytosis into control cells 86% was readily metabolized to galactosylceramide, ceramide, and stearic acid, which was reutilized in the synthesis of the major lipids found in cultured fibroblasts. Uptake and metabolism of the [14C]CS into cells from typical and atypical patients and carriers of metachromatic leukodystrophy (MLD), Krabbe disease, and Farber disease were observed....

Kudoh, Tooru; Wenger, David A.

1982-01-01

133

Specificity in the synergism between retinoic acid and EGF on the growth of adult human skin fibroblasts  

International Nuclear Information System (INIS)

Vitamin A (retinol) and five retinoids were tested for their ability to enhance epidermal growth factor (EGF) stimulation of adult human skin fibroblast growth in vitro. The retinoids utilized in this study were RO-1-5488 (all-trans-retinoic acid), RO-4-3780 (13-cis-retinoic acid), RO-10-9359, RO-10-1670, and RO-21-6583. Retinol and each retinoid were capable of stimulating fibroblast growth alone (0-86%), while 13-cis and all-trans-retinoic acid were the most potent in potentiating the EGF promotion of fibroblast growth. Since retinoic acid might enhance the EGF stimulation of cell growth by increasing either EGF receptor number or binding affinity, the binding of 125I-labeled EGF was carried out in the presence of retinoic acid and the data were subjected to a Scatchard-type analysis. No change in EGF receptor number or affinity was seen in the presence of retinoic acid. The data indicate a specific interaction between retinoid acid and EGF which results in the potentiation of the EGF-stimulated cell growth. Furthermore, the mechanism of this interaction does not seem to involve the initial binding of EGF to its plasma membrane receptor or the available number of EGF receptors located on the cell surface

134

Genome-wide differential gene expression in immortalized DF-1 chicken embryo fibroblast cell line  

OpenAIRE

Abstract Background When compared to primary chicken embryo fibroblast (CEF) cells, the immortal DF-1 CEF line exhibits enhanced growth rates and susceptibility to oxidative stress. Although genes responsible for cell cycle regulation and antioxidant functions have been identified, the genome-wide transcription profile of immortal DF-1 CEF cells has not been previously reported. Global gene expression in primary CEF and DF-1 cells was performed using a 4X44K chicken oligo mic...

Kong Byung-Whi; Lee Jeong Yoon; Bottje Walter G; Lassiter Kentu; Lee Jonghyuk; Foster Douglas N

2011-01-01

135

[Retraction] Generation of induced pluripotent stem cells using skin fibroblasts from patients with myocardial infarction under feeder-free conditions.  

Science.gov (United States)

After the publication of the article, the authors decided they wished to retract their manuscript for the following reasons. We wish to retract our research article entitled 'Generation of induced pluripotent stem cells using skin fibroblasts from patients with myocardial infarction under feeder-free conditions' published on the Molecular Medicine Reports 9: 837-842, 2014. In this article, we generated human iPSCs from skin fibroblasts from myocardial infarction patients in feeder-independent conditions. However, in subsequent researches, all of the cells generated and believed to be iPSCs showed negative expression of the pluripotent markers, Nanog and Rex1, and the cell surface marker, SSEA-1 and SSEA-4. Therefore we think the established iPS cells might not be real pluripotent stem cells. Based on the above mentioned, we ascertained that there must have some serious disadvantages in our design of experiment fundamentally. As a result, all authors involved unanimously agreed to retract this article and redesign our experiment. We deeply apologize to the readers for any inconvenience caused by this retraction. [the original article was published in the Molecular Medicine Reports 9: 837-842, 2014 DOI: 10.3892/mmr.2014.1885]. PMID:24866102

Li, Jun-Quan; Cheng, Ming; Tian, Wei-Chen; Zhang, Jian

2014-08-01

136

Inhibition of Oxidative Stress by Low-Molecular-Weight Polysaccharides with Various Functional Groups in Skin Fibroblasts  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was to evaluate the in cellulo inhibition of hydrogen-peroxide-induced oxidative stress in skin fibroblasts using different low-molecular-weight polysaccharides (LMPS prepared from agar (LMAG, chitosan (LMCH and starch (LMST, which contain various different functional groups (i.e., sulfate, amine, and hydroxyl groups. The following parameters were evaluated: cell viability, intracellular oxidant production, lipid peroxidation, and DNA damage. Trolox was used as a positive control in order to allow comparison of the antioxidant efficacies of the various LMPS. The experimentally determined attenuation of oxidative stress by LMPS in skin fibroblasts was: LMCH > LMAG > LMST. The different protection levels of these LMPS may be due to the physic-chemical properties of the LMPS’ functional groups, including electron transfer ability, metal ion chelating capacities, radical stabilizing capacity, and the hydrophobicity of the constituent sugars. The results suggest that LMCH might constitute a novel and potential dermal therapeutic and sun-protective agent.

Gregor P. C. Drummen

2013-09-01

137

Comparative proteomic analysis of extracellular matrix proteins secreted by two types of skin fibroblasts.  

Science.gov (United States)

The hair follicle dermal papilla is composed primarily of extracellular matrix (ECM) proteins secreted by resident fibroblasts. Dermal papilla is endowed with hair morphogenic properties, yet its composition is poorly characterized. In an attempt to understand its specificity better, we compared the protein composition of ECM secreted by cultured dermal papilla fibroblasts with that of dermal fibroblasts. ECM proteins are generally large, difficult to solubilize, and abundantly post-translationally modified. We thus implemented an original protocol for analyzing them: ECM samples were enzymatically digested directly in the culture flasks and analyzed by LC-MS/MS. Sequencing of proteolytic peptides by MS/MS yielded protein identification. The relative abundance of a given protein in dermal fibroblast versus dermal papilla samples was estimated by comparing proteolytic peptide intensities detected by MS. Using this approach, several matrix proteins were found to be present at markedly different levels in each ECM type; in particular, thrombospondin 1 and fibronectin appeared to be overrepresented in the dermal papilla fibroblast ECM. MS results were supported by Western blot and immunostaining experiments. In addition, peptide intensities were processed in two ways, which proved to favor either the quantification accuracy or the information precision at the sequence level. PMID:17068760

Pflieger, Delphine; Chabane, Sandrine; Gaillard, Olivier; Bernard, Bruno Alain; Ducoroy, Patrick; Rossier, Jean; Vinh, Joëlle

2006-11-01

138

Effect of low-dose-rate irradiation on the division potential of cells in vitro. V. Human skin fibroblasts from donors with a high risk of cancer  

International Nuclear Information System (INIS)

Skin fibroblasts from normal donors, donors with ataxia-telanglectasia or Fanconi's anemia, and from 1 cancer patient were treated with repeated ? radiation at about 16 rads per hour. The remaining division potential of all fibroblasts, except for the Fanconi's anemia cells, was reduced to different extents by radiation. The growth potential of Fanconl's anemia cells was increased in all the irradiated cultures. The increase was 54% in the group that survived the longest. These results were identical to those obtained with fibroblasts from certain species that have a high probability of transformation

139

Demonstration of 26-hydroxylation of C27-steroids in human skin fibroblasts, and a deficiency of this activity in cerebrotendinous xanthomatosis.  

OpenAIRE

26-Hydroxylation of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol and other C27-steroids was demonstrated in cultured skin fibroblasts from healthy individuals. Activities in skin fibroblasts were approximately 5-10% of those previously found in human liver homogenates, and were inhibited by CO. The apparent Km was lowest for 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol (1.3 mumol/liter) and highest for 5-cholestene-3 beta, 7 alpha-diol (12 mumol/liter). The rate of 26-hydroxylatio...

Skrede, S.; Bjo?rkhem, I.; Kvittingen, E. A.; Buchmann, M. S.; Lie, S. O.; East, C.; Grundy, S.

1986-01-01

140

Cholesteatoma fibroblasts promote epithelial cell proliferation through overexpression of epiregulin.  

Science.gov (United States)

To investigate whether keratinocytes proliferate in response to epiregulin produced by subepithelial fibroblasts derived from middle ear cholesteatoma. Tissue samples were obtained from patients undergoing tympanoplasty. The quantitative polymerase chain reaction and immunohistochemistry were performed to examine epiregulin expression and localization in cholesteatoma tissues and retroauricular skin tissues. Fibroblasts were cultured from cholesteatoma tissues and from normal retroauricular skin. These fibroblasts were used as feeder cells for culture with a human keratinocyte cell line (PHK16-0b). To investigate the role of epiregulin in colony formation by PHK16-0b cells, epiregulin mRNA expression was knocked down in fibroblasts by using short interfering RNA and epiregulin protein was blocked with a neutralizing antibody. Epiregulin mRNA expression was significantly elevated in cholesteatoma tissues compared with that in normal retroauricular skin. Staining for epiregulin was more intense in the epithelial cells and subepithelial fibroblasts of cholesteatoma tissues than in retroauricular skin. When PHK16-0b cells were cultured with cholesteatoma fibroblasts, their colony-forming efficiency was 50% higher than when these cells were cultured with normal skin fibroblasts. Also, knockdown of epiregulin mRNA in cholesteatoma fibroblasts led to greater suppression of colony formation than knockdown in skin fibroblasts. Furthermore, the colony-forming efficiency of PHK16-0b cells was significantly reduced after treatment with an epiregulin neutralizing antibody in co-culture with cholesteatoma fibroblasts, but not in co-culture with skin fibroblasts. These results suggest that keratinocyte hyperproliferation in cholesteatoma is promoted through overexpression of epiregulin by subepithelial fibroblasts via epithelial-mesenchymal interactions, which may play a crucial role in the pathogenesis of middle ear cholesteatoma. PMID:23826119

Yoshikawa, Mamoru; Kojima, Hiromi; Yaguchi, Yuichiro; Okada, Naoko; Saito, Hirohisa; Moriyama, Hiroshi

2013-01-01

141

Establishment of a novel immortalized cell line from ataxia telangiectasia fibroblasts and its use for the chromosomal assignment of radiosensitivity gene  

International Nuclear Information System (INIS)

An immortalized cell line was established from a female ataxia telangiectasia (AT) patient by the transfection of primary skin fibroblasts with origin-defective SV40 DNA. The cell line was characterized by a hypodiploid chromosome constitution and radiation hypersensitivity. The established cell line was used as a recipient for microcell-mediated chromosome transfer. Among seven G418-resistant clones obtained by the fusion with microcells from mouse A9 cells carrying a pSV2neo-tagged normal human chromosome 11, three clones showed restoration of radiation resistance with concomitant gain of an extra intact chromosome 11, while the others contained no recognizable or deleted chromosome 11. The association of the presence of 11q14?qter region with the radioresistance suggests the presence of AT gene in this chromosomal region. (author)

142

Cyclic phosphatidic acid and lysophosphatidic acid induce hyaluronic acid synthesis via CREB transcription factor regulation in human skin fibroblasts.  

Science.gov (United States)

Cyclic phosphatidic acid (cPA) is a naturally occurring phospholipid mediator and an analog of the growth factor-like phospholipid lysophosphatidic acid (LPA). cPA has a unique cyclic phosphate ring at the sn-2 and sn-3 positions of its glycerol backbone. We showed before that a metabolically stabilized cPA derivative, 2-carba-cPA, relieved osteoarthritis pathogenesis in vivo and induced hyaluronic acid synthesis in human osteoarthritis synoviocytes in vitro. This study focused on hyaluronic acid synthesis in human fibroblasts, which retain moisture and maintain health in the dermis. We investigated the effects of cPA and LPA on hyaluronic acid synthesis in human fibroblasts (NB1RGB cells). Using particle exclusion and enzyme-linked immunosorbent assays, we found that both cPA and LPA dose-dependently induced hyaluronic acid synthesis. We revealed that the expression of hyaluronan synthase 2 messenger RNA and protein is up-regulated by cPA and LPA treatment time dependently. We then characterized the signaling pathways up-regulating hyaluronic acid synthesis mediated by cPA and LPA in NB1RGB cells. Pharmacological inhibition and reporter gene assays revealed that the activation of the LPA receptor LPAR1, Gi/o protein, phosphatidylinositol-3 kinase (PI3K), extracellular-signal-regulated kinase (ERK), and cyclic adenosine monophosphate response element-binding protein (CREB) but not nuclear factor ?B induced hyaluronic acid synthesis by the treatment with cPA and LPA in NB1RGB cells. These results demonstrate for the first time that cPA and LPA induce hyaluronic acid synthesis in human skin fibroblasts mainly through the activation of LPAR1-Gi/o followed by the PI3K, ERK, and CREB signaling pathway. PMID:24845645

Maeda-Sano, Katsura; Gotoh, Mari; Morohoshi, Toshiro; Someya, Takao; Murofushi, Hiromu; Murakami-Murofushi, Kimiko

2014-09-01

143

Pyrophosphohydrolase activity and inorganic pyrophosphate content of cultured human skin fibroblasts. Elevated levels in some patients with calcium pyrophosphate dihydrate deposition disease.  

OpenAIRE

In calcium pyrophosphate dihydrate (CPPD) crystal deposition disease, metabolic abnormalities favoring extracellular inorganic pyrophosphate (PPi) accumulation have been suspected. Elevations of intracellular PPi in cultured skin fibroblasts from a single French kindred with familial CPPD deposition (19) and elevated nucleoside triphosphate pyrophosphohydrolase activity (NTPPPH), which generates PPi in extracts of CPPD crystal-containing cartilages (14) favor this suspicion. To determine whet...

Ryan, L. M.; Wortmann, R. L.; Karas, B.; Lynch, M. P.; Mccarty, D. J.

1986-01-01

144

ANALYSIS OF DNA DAMAGE AND REPAIR IN SKIN FIBROBLASTS OF INFANT AND OLDER CHILDREN USING THE IN VITRO ALKALINE COMET ASSAY  

Science.gov (United States)

ANALYSIS OF DNA DAMAGE AND REPAIR IN SKIN FIBROBLASTS OF INFANT AND OLDER CHILDREN USING THE IN VITRO ALKALINE COMET ASSAY, Alan H. Tennant1, Geremy W. Knapp1 and Andrew D. Kligerman1, 1Environmental Carcinogenesis Division, National Health and Environmental Effects Research Lab...

145

Whole venom of Loxosceles similis activates caspases-3, -6, -7, and -9 in human primary skin fibroblasts.  

Science.gov (United States)

Spiders of the Loxosceles genus represent a risk to human health due to the systemic and necrotic effects of their bites. The main symptoms of these bites vary from dermonecrosis, observed in the majority of cases, to occasional systemic hemolysis and coagulopathy. Although the systemic effects are well characterized, the mechanisms of cell death triggered by the venom of these spiders are poorly characterized. In this study, we investigated the cell death mechanisms induced by the whole venom of the spider Loxosceles similis in human skin fibroblasts. Our results show that the venom initiates an apoptotic process and a caspase cascade involving the initiator caspase-9 and the effector caspases-3, -6, and -7. PMID:24726468

Dantas, Arthur Estanislau; Horta, Carolina Campolina Rebello; Martins, Thais M M; do Carmo, Anderson Oliveira; Mendes, Bárbara Bruna Ribeiro de Oliveira; Goes, Alfredo M; Kalapothakis, Evanguedes; Gomes, Dawidson A

2014-06-01

146

Alterations in oncogene expression and radiosensitivity in the most frequently used SV40-transformed human skin fibroblasts  

International Nuclear Information System (INIS)

In comparison with primary cell cultures, SV40-transformed human skin fibroblasts, either from healthy donors or from patients suffering from ataxia-telangiectasia (AT) or xeroderma pigmentosum, are more resistant to the cytotoxic action of low LET 60cobalt ?-rays as well as to high LET ?-particles. Resistance factors calculated from D10's lie between 1.4 and 2.0. Northern blot analysis reveals spontaneous overexpression of the oncogenes c-myc, Ki-ras and c-raf and of the tumour suppressor gene p53 as a consequence of SV40 transformation. For c-myc, the increased expression is due to gene amplification and gene rearrangement. An even further increase in the expression of c-myc has been found for AT cells (AT5BI-VA) after moderate doses of 60cobalt ?-irradiation. A possible correlation between SV40-induced changes in gene expression and cellular radioresistance is discussed. (Author)

147

Cyclobutane-type pyrimidine photodimer formation and induction of ornithine decarboxylase in human skin fibroblasts after UV irradiation  

International Nuclear Information System (INIS)

Cyclobutane-type pyrimidine photodimers as well as the induction of ornithine decarboxylase (ODC) may serve as biochemical markers of the mutagenic and carcinogenic effects of ultraviolet light (UV). For this reason, it is important to compare the formation of pyrimidine dimers with the induction of ODC in human skin fibroblasts after irradiation with UVC (200-290 nm) and UVB (290-320 nm). In our studies we determined cytosine-thymine (C-T) as well as thymine-thymine dimer yields (T-T) by high-pressure liquid chromatography in cultures of neonatal normal human foreskin-derived fibroblasts after irradiation with UVC and UVB light. It was found that the yield of dimerization and the ratio of T-T/C-T decreased from the UVC to the UVB region. Time-course studies of ODC-induction in the same cells indicated that the maximal activity after UVB irradiation was retarded compared to UVC exposure. For the UV-induced ODC-levels, however, no significant difference in maximal induction could be measured after UVC and UVB irradiation at fluences where comparable yields of thymine dimerization are produced. Similar ODC-maxima were obtained with strains from children, while cells from adults showed significantly less pronounced ODC induction, indicating that ODC-response decreases with age and may therefore be used as a marker of aging

148

The influence of fluoride on the expression of inhibitors of Wnt/?-catenin signaling pathway in rat skin fibroblast Cells.  

Science.gov (United States)

The effective therapy of fluoride-induced bone diseases requires an understanding of the mechanism of the disorders. Changes in the inhibitors of the Wnt/?-catenin pathway, Dickkopf-1 (Dkk-1) and Sclerostin (SOST),were studied in supernatants harvested from rat skin fibroblasts cultured with varied doses of fluoride. The contents of SOST and Dkk-1 in fibroblast supernatants were assessed at four exposure time-points and investigated by using the method of ELISA. Compared to the relevant controls(0 mg F(?)/L), a significant decrease of the concentrations of SOST and Dkk-1 was observed as the fluoride concentration increased. Compared to the relevant time controls (24 h), a significant decrease of the concentrations of SOST and Dkk-1 was observed with the extension of time. Our results suggest that the Wnt/?-catenin pathway inhibitors Dkk-1 and SOST play an important role in skeletal fluorosis. They can be used as important indications for diagnosing bone metabolism changes caused by fluoride exposure and therapeutic targets in diseases resulting from fluoride exposure. PMID:22290293

Liu, Xiao-Li; Li, Chang-Cheng; Liu, Ke-Jian; Cui, Cai-Yan; Zhang, Yu-Zeng; Liu, Yun

2012-07-01

149

Immunomodulatory Effects of Bee Venom in Human Synovial Fibroblast Cell Line  

Science.gov (United States)

As in Iranian traditional medicine, bee venom (BV) is a promising treatment for the rheumatoid arthritis (RA) which is considered as a problematic human chronic inflammatory disease in the present time. Smoking is considered to be a major risk factor in RA onset and severity. The main aim of this study is to investigate the effects of BV on cigarette smoke-induced inflammatory response in fibroblast-like synoviocytes (FLS). Cytotoxicity of cigarette smoke condensate (CSC) and bee venom were determined by the tetrazolium (MTT) method in cultured synovial fibroblastes. The expression of interleukin-1? and sirtuin1 mRNA were analyzed by SYBR green real-time quantitative PCR. Differences between the mean values of treated and untreated groups were assessed by student t-test. Based on MTT assay, CSC and BV did not exert any significant cytotoxic effects up to 40 µg/mL and 10 µg/mL, respectively. Our results showed that interleukin-1? mRNA level was significantly up-regulated by CSC treatments in LPS-stimulated synoviocytes in a dose-dependent manner. Conversely, the expressions of IL-1? and Sirt1 were up-regulated even in lower concentrations of BV and attenuated at higher concentrations. Also, BV attenuated the CSC-induced and LPS-induced inflammatory responses in synovial fibroblasts. Our results support the epidemiological studies indicating pro-inflammatory effects of CSC and anti-inflammatory effects of BV on FLS cell line.

Mohammadi, Ebrahim; Vatanpour, Hossein; H Shirazi, Farshad

2015-01-01

150

Effects of CO2 laser irradiation on the wettability and human skin fibroblast cell response of magnesia partially stabilised zirconia  

International Nuclear Information System (INIS)

Human skin fibroblast cells in vitro responses on the surface of a bioinert zirconia ceramic partially stabilised with magnesia partially stabilised zirconia (MgO-PSZ) bioinert ceramic before and after CO2 laser treatment were investigated to find the interrelationship between the cell adhesion, wettability and laser parameters. Contact angle, ?, measurements of a set of test liquids were a clear indication that surface treatment of the MgO-PSZ with a CO2 laser brought about a reduction in ?, indicating that the wettability of the MgO-PSZ had been enhanced. A relationship was found between the wettability and the microstructure of the MgO-PSZ surface and laser processing parameters. It was subsequently deduced that the factors active in causing the observed modification in the wettability of the MgO-PSZ were the increases in the surface O2 content and the polar component of the surface energy, ?svp, the latter resulting from surface melting and resolidification. Moreover, the investigation into the human skin fibroblast cell response revealed that the CO2 laser treatment of the MgO-PSZ had resulted in a surface favourable for cell adhesion, as the extent of cell attachment and adhesion on the MgO-PSZ surface was enhanced depending on laser parameters. Such an improvement in cell adhesion, which could be greatly beneficial to developing enhanced bonding at the tissue and implant interface, was influenssue and implant interface, was influenced by the surface properties of the modified MgO-PSZ, particular wettability

151

Cholesterol Metabolism in Brain and Skin Fibroblasts from Sarda Breed Sheep With Scrapie-resistant and Scrapie-susceptible Genotypes  

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Full Text Available Scrapie is a fatal spongiform encephalopathy of sheep, a transmissible form of prion disease caused by neuronal accumulation of the aberrantly conformed prion protein (PrPsc. Currently, no ante-mortem diagnostic tests are available to detect this untreatable disease in the pre-clinical stage, thus making difficult to control its spread. Recent evidence suggests that the production of PrPsc can be modulated by the levels of membrane cholesterol in neuronal cells. Since cholesterol levels in cell membranes are dependent on cholesterol homeostasis in the whole organism, we studied cholesterol metabolism in brain tissues, plasma and skin fibroblasts of Sarda breed sheep with scrapie-resistant (ARR/ARR and scrapie-susceptible (ARQ/ARQ prion protein genotypes, both not infected (ARQ/ARQ- and infected (ARQ/ARQ+ with scrapie. We found that, the levels of cytoplasmic cholesterol esters (CE in brains and skin fibroblasts from sheep with the ARQ/ARQ genotype were consistently higher than those from sheep with the ARR/ARR genotype. Conversely, the levels of free cholesterol (FC were lower in ARQ/ARQ, as compared to ARR/ARR sheep, thus resulting in a sharp reduction of the FC/CE ratio. Moreover, both uninfected and infected ARQ/ARQ sheep showed abnormally low levels of high density lipoprotein-cholesterol (HDL-C in their plasma, as compared to ARR/ARR sheep. These data other than adding new strength to the notion that altered levels of intracellular cholesterol may indicate the presence of a lipid metabolic state that predisposes to infection with, and accumulation of, PrPsc in the brain, discriminate for the first time between two distinct but related cellular pools of cholesterol, namely membrane FC on one hand and cytoplasmic CE on the other.

Alessandra Pani

2007-01-01

152

Wound healing morbidity in STS patients treated with preoperative radiotherapy in relation to in vitro skin fibroblast radiosensitivity, proliferative capacity and TGF-? activity  

International Nuclear Information System (INIS)

Background and purpose: In a recent study, we demonstrated that the ability of dermal fibroblasts, obtained from soft tissue sarcoma (STS) patients, to undergo initial division in vitro following radiation exposure correlated with the development of wound healing morbidity in the patients following their treatment with preoperative radiotherapy. Transforming growth factor beta (TGF-?) is thought to play an important role in fibroblast proliferation and radiosensitivity both of which may impact on wound healing. Thus, in this study we examined the interrelationship between TGF-? activity, radiosensitivity and proliferation of cultured fibroblasts and the wound healing response of STS patients after preoperative radiotherapy to provide a validation cohort for our previous study and to investigate mechanisms. Patients and methods: Skin fibroblasts were established from skin biopsies of 46 STS patients. The treatment group consisted of 28 patients who received preoperative radiotherapy. Eighteen patients constituted a control group who were either irradiated postoperatively or did not receive radiation treatment. Fibroblast cultures were subjected to the colony forming and cytokinesis-blocked binucleation assays (low dose rate: ?0.02 Gy/min) and TGF-? assays (high dose-rate: ?1.06 Gy/min) following ?-irradiation. Fibroblast radiosensitivity and initial proliferative ability were represented by the surviving fraction at 2.4 Gy (SF2.4) and binucleation insub>2.4) and binucleation index (BNI), respectively. Active and total TGF-? levels in fibroblast cultures were determined using a biological assay. Wound healing complication (WHC), defined as the requirement for further surgery or prolonged deep wound packing, was the clinical endpoint examined. Results: Of the 28 patients treated with preoperative radiotherapy, 8 (29%) had wound healing difficulties. Fibroblasts from patients who developed WHC showed a trend to retain a significantly higher initial proliferative ability after irradiation compared with those from individuals in the treatment group with normal wound healing, consistent with the results of our previous study. No link was observed between fibroblast radiosensitivity and WHC. Neither active nor total TGF-? levels in cultures were significantly affected by irradiation. Fibroblast proliferation in unirradiated and irradiated cultures, as well as radiosensitivity, was not influenced by TGF-? content. TGF-? expression in fibroblast cultures did not reflect wound healing morbidity. Conclusions: These data are consistent with our previous study and combined the results suggest that in vitro fibroblast proliferation after irradiation may be a useful predictor of wound healing morbidity in STS patients treated with preoperative radiotherapy. TGF-? levels in culture do not predict WHC, suggesting that the role of TGF-? in wound healing is likely controlled by other in vivo factors

153

Development of Biomimetic Tilapia Collagen Nanofibers for Skin Regeneration through Inducing Keratinocytes Differentiation and Collagen Synthesis of Dermal Fibroblasts.  

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In this study, tilapia skin collagen sponge and electrospun nanofibers were developed for wound dressing. The collagen sponge was composed of at least two ?-peptides, and its denaturation temperature was 44.99 °C. It did not change the number of spleen-derived lymphocytes in BALB/c mice, the ratio of CD4(+)/CD8(+) lymphocytes, and the level of IgG or IgM in Sprague-Dawley rat. The contact angle, tensile strength, and weight loss temperature of collagen nanofibers were 21.2°, 6.72 ± 0.44 MPa, and 300 °C, respectively. The nanofibers could promote the viabilities of human keratinocytes (HaCaTs) and human dermal fibroblasts (HDFs), inducing epidermal differentiation through the gene expression of involucrin, filaggrin, and type I transglutaminase of HaCaTs, and they could also accelerate migration of HaCaTs with the expression of matrix metalloproteinase-9 and transforming growth factor-?1 (TGF-?1). Besides, the nanofibers could upregulate the protien level of Col-I in HDFs both via a direct effect and TGF-?1 secreted from HaCaTs, thus facilitating the formation of collagen fibers. Furthermore, the collagen nanofibers stimulated the skin regeneration rapidly and effectively in vivo. These biological effects could be explained as the contributions from the biomimic extracellular cell matrix structure, hydrophilicity, and the multiple amino acids of the collagen nanofibers. PMID:25598076

Zhou, Tian; Wang, Nanping; Xue, Yang; Ding, Tingting; Liu, Xin; Mo, Xiumei; Sun, Jiao

2015-02-11

154

Effects of hyperthermia and ionizing radiation in normal and ataxia telangiectasia human fibroblast lines  

International Nuclear Information System (INIS)

The effects of 450C hyperthermia and ? radiation have been studied in three normal human fibroblast lines (GM38, GM730, WI38) and compared to the effects in two lines derived from patients with the hereditary disease ataxia telangiectasia (AR3BI, AT5BI). All lines, both normal and ?-sensitive AT, showed a similar resistance to killing by heat alone, suggesting that the defect responsible for the increased radiation sensitivity in AT lines does not confer increased heat sensitivity. Shouldered survival curves were obtained in each case indicating the ability to accumulate sublethal heat damage. All normal and AT cell lines exhibited increased resistance to the lethal effects of heat in response to a thermal stress, indicating that the defect that causes radiosensitivity in AT cell lines does not prevent the induction of thermotolerance. It was hypothesized that in normal cells, this heat treatment inactivates the process which is already defective in AT lines, and that this process may be required for the proper rejoining of double-strand breaks produced during the repair of other radiation-induced lesions

155

In vitro study for laser gene transfer in BHK-21 fibroblast cell line  

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Modifications to our previously introduced system for laser microbeam cell surgery were carried out in the present work to match animal cells. These modifications included: 1- Using other laser system that used before, Excimer laser with 193 and 308 nm wavelengths. The used laser here, is He-Cd with low power and 441.5 nm wavelength in the visible region. 2- Instead of using pulsed laser, we used here CW He-Cd chopped by electrical chopper, which is synchronized with the mechanical motion of the mobile stage with step 40 microns, according to cell dimensions to avoid puncturing the same cell twice. The advantages of the modified here laser setup for gene transfer is: it is less damaging to the sensitive animal cell which has thin cell membrane. The present work aimed to: 1- Design a modified laser microbeam cell surgery, applicable to animal cells, such as fibroblast cells 2- To examine the efficiency of such system. 3- To assure gene transfer and its expression in the used cells. 4- To evaluate the ultra damages produced from using the laser beam as a modality for gene transfer. On the other wards, to introduce: safe, efficient and less damaging modality for gene transfer in animal cells. To achieve these goals, we applied the introduced here home-made laser setup with its synchronized parameters to introduce pBK-CMV phagemid, containing LacZ and neomycin resistance (neor )genes into BHK-21 fibroblast cell line. The results of the present work showed that: 1- Our modified laser microbeam cell surgery setup proved to be useful and efficient tool for gene transfer into fibroblast cells. 2- The presence and expression of LacZ gene was achieved using histochemical LacZ assay. 3- Selection of G418 antibiotic sensitivity assay confirmed the presence and expression towards stability of neor gene with time. 4- Presence of LacZ and neor genes in the genomic DNA of transfected fibroblast cells was indicated using PCR analysis. 5- Transmission electron microscopy indicated that, no ultradamages or changes for cell; membrane, organilles or any component of transfected fibroblast cell as a result of using laser microbeam compared with control cell.

Abdel Aziz, M.; Salem, D. S.; Salama, M. S.; Badr, Y.

2009-02-01

156

The Effects of Macrophage-Stimulating Protein on the Migration, Proliferation and Collagen Synthesis of Skin Fibroblasts in Vitro and in Vivo.  

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Macrophage-stimulating protein (MSP), an important cytokine with multiple functions, is highly expressed in adipose mesenchymal stem cells-conditioned medium (ASC-CM). ASCs can effectively promote wound healing through paracrine mechanism, suggesting that MSP may play a critical role in wound healing. Through binding to its receptor, RON, it can activate epithelial cells and work as an inflammatory mediator. In this study, we found RON was also expressed on dermal fibroblasts and investigated the effects of MSP on proliferation, migration and collagen synthesis of fibroblasts. With the treatment of different concentrations of MSP (0, 1, 10, 20, 50, and 100ng/ml) on fibroblasts, proliferation, migration and collagen synthesis were analyzed by CCK-8, transwell and RT-PCR. Under the treatment of MSP, the migration, Collagen I ,?synthesis and MMP1 mRNA expression of fibroblasts were up-regulated significantly, although there was no effect on fibroblasts proliferation, and the optimal concentration of MSP for migration and collagen synthesis was 10ng/ml. In the in vivo study, 10ng/ml MSP was applied to full-thickness skin wound with bacterial cellulose membranes, and this treatment could accelerate the wound healing rate and increased the collagen synthesis of wound sites. This study suggested that MSP appears to promote the migration of fibroblasts, enhances collagen synthesis and remodeling, and to effectively improve wound healing. PMID:25315688

Zhao, Jiajia; Hu, Li; Gong, Niya; Tang, Qingming; Du, Luyang; Chen, Lili

2014-10-14

157

Photochemical damage to skin fibroblasts caused by protoporphyrin and violet light  

International Nuclear Information System (INIS)

Foreskin fibroblasts cultured in a medium containing protoporphyrin and exposed to violet light lose the capacity to proliferate. This phenomenon can be assessed on the basis of the ability of the irradiated cells to form colonies. Potentially lethal injuries can, however, be repaired during post-irradiation incubation under optimal growth conditions. We investigated the photodynamically induced transformations of certain molecular targets in the irradiated cells. Biochemical analysis showed that only traces of unsaturated fatty acids were oxidized, but SH groups of both the membranes and the cytosol appeared to be very sensitive targets. Of the tryptophan content, 20% was damaged during irradiation. Recovery was observed during post-irradiation incubation. The tryptophan content and the SH groups recovered to some extent, and these results showed a good correlation with the regeneration of surviving cells. (orig.)

158

Phosphoproteomics profiling of human skin fibroblast cells reveals pathways and proteins affected by low doses of ionizing radiation  

Energy Technology Data Exchange (ETDEWEB)

Background: High doses of ionizing radiation result in biological damage, however the precise relationships between long term health effects, including cancer, and low dose exposures remain poorly understood and are currently extrapolated using high dose exposure data. Identifying the signaling pathways and individual proteins affected at the post-translational level by radiation should shed valuable insight into the molecular mechanisms that regulate dose dependent responses to radiation. Principle Findings: We have identified 6845 unique phosphopeptides (2566 phosphoproteins) from control and irradiated (2 and 50 cGy) primary human skin fibroblasts one hour post-exposure. Dual statistical analyses based on spectral counts and peak intensities identified 287 phosphopeptides (from 231 proteins) and 244 phosphopeptides (from 182 proteins) that varied significantly following exposure to 2 and 50 cGy respectively. This screen identified phosphorylation sites on proteins with known roles in radiation responses including TP53BP1 as well as previously unidentified radiation responsive proteins such as the candidate tumor suppressor SASH1. Bioinformatics analyses suggest that low and high doses of radiation affect both overlapping and unique biological processes and suggest a role of MAP kinase and protein kinase A (PKA) signaling in the radiation response as well as differential regulation of p53 networks at low and high doses of radiation. Conlcusions: Our results represent the most comprehensive analysis of the phosphoproteomes of human primary fibroblasts exposed to multiple doses of ionizing radiation published to date and provides a basis for the systems level identification of biological processes, molecular pathways and individual proteins regulated in a dose dependent manner by ionizing radiation. Further study of these modified proteins and affected networks should help to define the molecular mechanisms that regulate biological responses to radiation at different radiation doses and elucidate the impact of low dose radiation exposure on human health.

Yang, Feng; Waters, Katrina M.; Miller, John H.; Gritsenko, Marina A.; Zhao, Rui; Du, Xiuxia; Livesay, Eric A.; Purvine, Samuel O.; Monroe, Matthew E.; Wang, Yingchun; Camp, David G.; Smith, Richard D.; Stenoien, David L.

2010-11-30

159

NADPH oxidase-2 is a key regulator of human dermal fibroblasts: a potential therapeutic strategy for the treatment of skin fibrosis.  

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The proliferation of human skin dermal fibroblasts (HDFs) is a critical step in skin fibrosis, and transforming growth factor-beta1 (TGF-?1) exerts pro-oxidant and fibrogenic effects on HDFs. In addition, the oxidative stress system has been implicated in the pathogenesis of skin disease. However, the role of NADPH oxidase as a mediator of TGF-?1-induced effects in HDFs remains unknown. Thus, our aim was to investigate the role of NADPH in human skin dermal fibroblasts. Primary fibroblasts were cultured and pretreated with various stimulants. Real-time Q-PCR and Western blotting analyses were used for mRNA and protein detection. In addition, siRNA technology was applied for gene knock-down analysis. Hydrogen peroxide production and 2',7'-dichlorofluorescein diacetate (DCFDA) measurement assay were performed. Here, our findings demonstrated that HDFs express key components of non-phagocytic NADPH oxidase mRNA. TGF-?1 induced NOX2 and reactive oxygen species formation via NADPH oxidase activity. In contrast, NOX3 was barely detectable, and other NOXs did not display significant changes. In addition, TGF-?1 phosphorylated MAPKs and increased activator protein-1 (AP-1) in a redox-sensitive manner, and NOX2 suppression inhibited baseline and TGF-?1-mediated stimulation of Smad2 phosphorylation. Moreover, TGF-?1 stimulated cell proliferation, migration, collagen I and fibronectin expression, and bFGF and PAI-1 secretion: these effects were attenuated by diphenylene iodonium (DPI), an NADPH oxidase inhibitor, and NOX2 siRNA. Importantly, NOX2 siRNA suppresses collagen production in primary keloid dermal fibroblasts. These findings provide the proof of concept for NADPH oxidase as a potential target for the treatment of skin fibrosis. PMID:24981855

Zhang, Guo-You; Wu, Liang-Cai; Dai, Tao; Chen, Shi-Yi; Wang, An-Yuan; Lin, Kang; Lin, Da-Mu; Yang, Jing-Quan; Cheng, Biao; Zhang, Li; Gao, Wei-Yang; Li, Zhi-Jie

2014-09-01

160

Establishment of ultra long-lived cell lines by transfection of TERT into normal human fibroblast TIG-1 and their characterization.  

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To establish useful human normal cell lines, TERT (telomerase reverse transcriptase) cDNA was transfected into normal female lung fibroblast, TIG-1. After long-term-sub-cultivation of 74 individual clones selected for resistance to G418, we obtained 55 cultures with normal range of life span [75 PDL (population doubling level)], 16 cultures with extended life span (75-140 PDL). In addition, 3 immortal cell strains and unexpectedly, one ultra long-lived cell line (ULT-1) with life span of 166 PDL were established. IMT-1, one of the immortal cell strains was confirmed to maintain long telomere length, high telomerase activity and an extremely low level of p16INK4A. They also showed moderate p53 and p21CIP1 expression, keeping vigorous growth rate even at 450 PDL. High level of fibronectin and collagen 1? expression confirmed IMT-1 as normal fibroblasts, although one X chromosome had been lost. ULT-1, however, kept a near normal karyotypes and had shortening of telomere length, high expression of p16INK4A, moderate levels of senescence associated-?-galactosidase positive cells and decreased growth rate only after 150 PDs (population doublings), and finally reached senescence at 166 PDL with morphology of normal senescent fibroblasts. As resources of standard normal human cell, abundant vials of early and middle passages of ULT-1 have been stocked. The use of the cell line is discussed, focusing on isograft of artificial skin and screening of anti-aging or safe chemical agents. PMID:22273270

Kamada, Mizuna; Kumazaki, Tsutomu; Matsuo, Taira; Mitsui, Youji; Takahashi, Tomoko

2012-06-01

161

[Human skin reconstructed in vitro as a model to study the keratinocyte, the fibroblast and their interactions: photodamage and repair processes].  

Science.gov (United States)

The protective role of the skin is provided by the two major compartments of the skin, dermis and epidermis. Both are affected in the long term by consequences of sun exposure such as skin photoaging and cancer development. Characterization of UV-induced skin response at cellular and molecular levels is needed for prevention or correction of these long term effects. The human skin reconstructed in vitro, comprising both a living dermal equivalent and a fully differentiated epidermis represents a predictive tool to characterize wavelength and cell type specific biological damage together with tissular distribution. While UVB directly affects epidermis, inducing DNA lesions and apoptotic sunburn keratinocytes, UVA radiation can directly target the dermal compartment through ROS generation, dermal fibroblasts alterations and extracellular matrix (ECM) modifications. Interactions between the two compartments have also been found, especially for MMP1 induction. In the normal population, photodamage can be repaired through specialized systems. Using skin cells from Xeroderma pigmentosum (XP, a photosensitive and cancer-prone disease), a DNA-repair deficient skin has been developed in vitro. Specific features due to intrinsic XP cell phenotype have been discovered, some of them being indicative of early steps of neoplasia and suggesting a particular role for stroma-epithelium interactions. Finally, human reconstructed skin can be used for approaches designed to regenerate photodamaged skin. The dermal-epidermal junction (DEJ), which is crucial for skin cohesion, is drastically altered in photo-aged skin. The three-dimensional skin model allowed to visualize the improving effects of vitamin C on the DEJ. Modified skin models, lacking one cell type, allowed us to determine the cellular origin of the different markers, their spatial localization, and the respective roles and interactions of keratinocytes and fibroblasts during DEJ formation. All together these studies give a global and tissular view concerning the effects of UV light on skin cells and emphazise the interest of such models for general aspects of cellular biology. By allowing the control of cells used to reconstruct the model and their origin, these studies make it possible to assess the respective role of the two major cellular actors of the skin as well as their interactions. Ongoing research about incorporating other cell types may certainly give rise to even more relevant models. PMID:16738525

Bernerd, Françoise

2005-01-01

162

Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite  

International Nuclear Information System (INIS)

We have shown that UVA (320-380 nm) radiation, hydrogen peroxide, and sodium arsenite induce a stress protein of approximately 32 kDa in human skin fibroblasts. The synthesis and cloning of cDNA from arsenite-induced mRNA populations have now allowed us to unequivocally identify the 32-kDa protein as heme oxygenase. By mRNA analysis we have shown that the heme oxygenase gene is also induced in cultured human skin fibroblasts by UVA radiation, hydrogen peroxide, cadmium chloride, iodoacetamide, and menadione. The known antioxidant properties of heme catabolites taken together with the observation of a high level of induction of the enzyme in cells from an organ not involved in hemoglobin breakdown strongly supports the proposal that the induction of heme oxygenase may be a general response to oxidant stress and constitutes an important cellular defense mechanism against oxidative damage

163

Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite  

Energy Technology Data Exchange (ETDEWEB)

We have shown that UVA (320-380 nm) radiation, hydrogen peroxide, and sodium arsenite induce a stress protein of approximately 32 kDa in human skin fibroblasts. The synthesis and cloning of cDNA from arsenite-induced mRNA populations have now allowed us to unequivocally identify the 32-kDa protein as heme oxygenase. By mRNA analysis we have shown that the heme oxygenase gene is also induced in cultured human skin fibroblasts by UVA radiation, hydrogen peroxide, cadmium chloride, iodoacetamide, and menadione. The known antioxidant properties of heme catabolites taken together with the observation of a high level of induction of the enzyme in cells from an organ not involved in hemoglobin breakdown strongly supports the proposal that the induction of heme oxygenase may be a general response to oxidant stress and constitutes an important cellular defense mechanism against oxidative damage.

Keyse, S.M.; Tyrrell, R.M.

1989-01-01

164

Application of a Novel Population of Multipotent Stem Cells Derived from Skin Fibroblasts as Donor Cells in Bovine SCNT  

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Undifferentiated stem cells are better donor cells for somatic cell nuclear transfer (SCNT), resulting in more offspring than more differentiated cells. While various stem cell populations have been confirmed to exist in the skin, progress has been restricted due to the lack of a suitable marker for their prospective isolation. To address this fundamental issue, a marker is required that could unambiguously prove the differentiation state of the donor cells. We therefore utilized magnetic activated cell sorting (MACS) to separate a homogeneous population of small SSEA-4+ cells from a heterogeneous population of bovine embryonic skin fibroblasts (BEF). SSEA-4+ cells were 8-10 ?m in diameter and positive for alkaline phosphatase (AP). The percentage of SSEA-4+ cells within the cultured BEF population was low (2-3%). Immunocytochemistry and PCR analyses revealed that SSEA-4+ cells expressed pluripotency-related markers, and could differentiate into cells comprising all three germ layers in vitro. They remained undifferentiated over 20 passages in suspension culture. In addition, cloned embryos derived from SSEA-4 cells showed significant differences in cleavage rate and blastocyst development when compared with those from BEF and SSEA-4? cells. Moreover, blastocysts derived from SSEA-4+ cells showed a higher total cell number and lower apoptotic index as compared to BEF and SSEA-4– derived cells. It is well known that nuclei from pluripotent stem cells yield a higher cloning efficiency than those from adult somatic cells, however, pluripotent stem cells are relatively difficult to obtain from bovine. The SSEA-4+ cells described in the current study provide an attractive candidate for SCNT and a promising platform for the generation of transgenic cattle. PMID:25602959

Pan, Shaohui; Chen, Wuju; Liu, Xu; Xiao, Jiajia; Wang, Yanqin; Liu, Jun; Du, Yue; Wang, Yongsheng; Zhang, Yong

2015-01-01

165

The effect of Centella asiatica, vitamins, glycolic acid and their mixtures preparations in stimulating collagen and fibronectin synthesis in cultured human skin fibroblast.  

Science.gov (United States)

Centella asiatica (Linn.) Urban is well known in promoting wound healing and provides significant benefits in skin care and therapeutic products formulation. Glycolic acid and vitamins also play a role in the enhancement of collagen and fibronectin synthesis. Here, we evaluate the specific effect of Centella asiatica (CA), vitamins, glycolic acid and their mixture preparations to stimulate collagen and fibronectin synthesis in cultured human fibroblast cells. The fibroblast cells are incubated with CA, glycolic acid, vitamins and their mixture preparations for 48 h. The cell lysates were analyzed for protein content and collagen synthesis by direct binding enzyme immunoassay. The fibronectin of the cultured supernatant was measured by sandwich enzyme immunoassay. The results showed that CA, glycolic acid, vitamins A, E and C significantly stimulate collagen and fibronectin synthesis in the fibroblast. Addition of glycolic acid and vitamins to CA further increased the levels of collagen and fibronectin synthesis to 8.55 and 23.75 ?g/100 ?g, respectively. CA, glycolic acid, vitamins A, E, and C, and their mixtures demonstrated stimulatory effect on both extra-cellular matrix synthesis of collagen and fibronectin in in vitro studies on human foreskin fibroblasts, which is beneficial to skin care and therapeutic products formulation. PMID:24577907

Hashim, Puziah

2014-03-01

166

The histamine degradative uptake pathway in human vascular endothelial cells and skin fibroblasts is dependent on extracellular Na+ and Cl-  

International Nuclear Information System (INIS)

We have previously reported that human vascular endothelial cells and skin fibroblasts carry out degradation of [3H]histamine by a mechanism involving two successive enzymatic steps: imidazole ring tele-methylation by the cells' endogenous methyltransferase and subsequent amine oxidation by an exogenous diamine oxidase. Both histamine and the exogenous second enzyme in the pathway associate with the cells via separate binding sites or receptors. The enzymatic degradation process results in cellular accumulation of the proximal and distal metabolites tele-methylhistamine and 1-methyl-4-imidazoleacetic acid (MIAA). We have now demonstrated that this two-stage histamine degradative pathway is dependent on Na+ and Cl- in the extracellular environment. Accumulation of [3H] histamine-derived products is partially inhibited under conditions of Na+ deprivation and more substantially when Cl- is also withdrawn. The individual tele-methylation and amine oxidation enzymatic reactions themselves are unaffected or actually facilitated under these conditions. This indicates that it is the cellular mechanism for uptake coupled to the degradative pathway which reflects the cation and anion dependency. Restoration of degradative uptake displays a biphasic Na+ concentration curve, suggesting that the uptake process may be driven by multiple components. These findings indicate a role for both inward Na+ and Cl- ion movement in this cellular degradative uptake mechanismadative uptake mechanism

167

Altered binding of 125I-labeled calmodulin to a 46.5-kilodalton protein in skin fibroblasts cultured from patients with cystic fibrosis  

International Nuclear Information System (INIS)

The levels of calmodulin and calmodulin-binding proteins have been determined in cultured skin fibroblasts from patients with cystic fibrosis (CF) and age- and sex-matched controls. Calmodulin ranged from 0.20 to 0.76 microgram/mg protein; there was no difference between calmodulin concentration in fibroblasts from CF patients and controls. Calmodulin-binding proteins of 230, 212, 204, 164, 139, 70, 59, 46.5, and 41 kD were identified. A protein with a mobility identical to the 59-kD calmodulin-binding protein was labeled by antiserum against calmodulin-dependent phosphatase. Although Ca2+/calmodulin-dependent phosphatase activity was detected, there was no different in activity between control and CF fibroblasts or in the level of phosphatase protein as determined by radioimmunoassay. Lower amounts of 125I-calmodulin were bound to the 46.5-kD calmodulin-binding protein in CF fibroblasts as compared with controls. The 46.5-kD calmodulin-binding protein may be reduced in CF fibroblasts or its structure may be altered resulting in a reduced binding capacity and/or affinity for calmodulin and perhaps reflecting, either directly or indirectly, the genetic defect responsible for cystic fibrosis

168

A study on the activity of fibroblast cells in connection with tissue recovery in the wounds of skin injury after whole-body irradiation  

International Nuclear Information System (INIS)

The 6 Gy of whole-body irradiation (WBI) with gamma rays results in an impairment of injured skin tissue recovery and renders a delay in the healing process. For an understanding of whether WBI has damaging effects on fibroblasts in wounds, fibroblasts in wounds combined with WBI and those of simple incision were isolated and cultivated, and abilities connected with tissue repair, including proliferation, attachment, adhesion, and apoptosis, were determined by direct cell count, immunohistochemical staining for proliferation cell nuclear antigen (PCNA), and TdT-mediated dUTP-biotin nick end-labeling (TUNEL) assay. The results showed that the abilities of proliferation and the attachment and adhesion of fibroblasts from wounds combined with WBI significantly decreased in comparison with those having simple incisions on the 3rd and 5th days of posttrauma, whereas the apoptotic ratio of fibroblasts from wounds combined with WBI significantly increased. These data suggest that WBI may exert damaging effects on fibroblasts in wounds, which might be one of the dominant reasons for the impaired healing of wounds combined with WBI. (author)

169

Modulation of procollagen gene expression by retinoids. Inhibition of collagen production by retinoic acid accompanied by reduced type I procollagen messenger ribonucleic acid levels in human skin fibroblast cultures.  

OpenAIRE

Recent clinical observations have suggested that retinoids, which are in frequent use in dermatology, can affect the connective tissue metabolism in skin and other tissues. In this study, we examined the effects of several retinoids on the metabolism of collagen by human skin fibroblasts in culture. Incubation of cultured fibroblasts with all-trans-retinoic acid or 13-cis-retinoic acid, in 10(-5) M or higher concentrations, markedly reduced the procollagen production, as measured by synthesis...

Oikarinen, H.; Oikarinen, A. I.; Tan, E. M.; Abergel, R. P.; Meeker, C. A.; Chu, M. L.; Prockop, D. J.; Uitto, J.

1985-01-01

170

Establishment of an immortal cynomolgus macaque fibroblast cell line for propagation of cynomolgus macaque cytomegalovirus (CyCMV).  

Science.gov (United States)

Cynomolgus macaques are widely used as an animal model in biomedical research. We have established an immortalized cynomolgus macaque fibroblast cell line (MSF-T) by transducing primary dermal fibroblasts isolated from a 13-year-old male cynomolgus macaque with a retrovirus vector expressing human telomerase reverse transcriptase (hTERT). The MSF-T cells showed increased telomerase enzyme activity and reached over 200 in vitro passages compared to the non-transduced dermal fibroblasts, which reached senescence after 43 passages. The MSF-T cell line is free of mycoplasma contamination and is permissive to the newly identified cynomolgus macaque cytomegalovirus (CyCMV). CyCMV productively infects MSF-T cells and induces down-regulation of MHC class I expression. The MSF-T cell line will be extremely useful for the propagation of CyCMV and other cynomolgus herspesviruses in host-derived fibroblast cells, allowing for the retention of host-specific viral genes. Moreover, this cell line will be beneficial for many in vitro experiments related to this animal model. PMID:23232747

Ambagala, Aruna P; Marsh, Angie K; Chan, Jacqueline K; Mason, Rosemarie; Pilon, Richard; Fournier, Jocelyn; Sandstrom, Paul; Willer, David O; MacDonald, Kelly S

2013-05-01

171

Comparison of surface fibroblastic cells in subcutaneous air pouch and synovial lining: differences in uridine diphosphoglucose dehydrogenase activity.  

OpenAIRE

In contrast to synovial tissue, rat subcutaneous air pouch lining was found to lack cells showing high activity of uridine diphosphoglucose dehydrogenase, an enzyme involved in hyaluronan synthesis. This indicates that the properties of cells on the surface of synovium are not determined simply by tissue cavitation. Shearing forces may be more important in inducing the specialized behaviour of synovial surface fibroblasts.

Wilkinson, L. S.; Moore, A. R.; Pitsillides, A. A.; Willoughby, D. A.; Edwards, J. C.

1993-01-01

172

Selection of reference genes for gene expression studies in ultraviolet B-irradiated human skin fibroblasts using quantitative real-time PCR  

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Full Text Available Abstract Background Reference genes are frequently used to normalise mRNA levels between different samples. The expression level of these genes, however, may vary between tissues or cells and may change under certain circumstances. Cytoskeleton genes have served as multifunctional tools for experimental studies as reference genes. Our previous studies have demonstrated that the expression of vimentin, one cytoskeletal protein, was increased in ultraviolet B (UVB-irradiated fibroblasts. Thus, we examined the expression of other cytoskeleton protein genes, ACTB (actin, beta, TUBA1A (tubulin, alpha 1a, and TUBB1 (tubulin, beta 1, in human dermal fibroblasts irradiated by UVB to determine which of these candidates were the most appropriate reference genes. Results Quantitative real-time PCR followed by analysis with the NormFinder and geNorm software programmes was performed. The initial screening of the expression patterns demonstrated that the expression of VIM was suppressed after UVB irradiation at doses ?25 mJ/cm2 and that the expression of TUBA1A was significantly reduced by UVB doses ?75 mJ/cm2 in cultured human dermal fibroblasts. The analysis of the experimental data revealed ACTB to be the most stably expressed gene, followed by GAPDH (aglyceraldehyde-3-phosphate dehydrogenase, under these experimental conditions. By contrast, VIM was found to be the least stable gene. The combination of ACTB and TUBB1 was revealed to be the gene pair that introduced the least systematic error into the data normalisation. Conclusion The data herein provide evidence that ACTB and TUBB1 are suitable reference genes in human skin fibroblasts irradiated by UVB, whereas VIM and TUBA1A are not and should therefore be excluded as reference genes in any gene expression studies involving UVB-irradiated human skin fibroblasts.

Qu Tao

2011-02-01

173

Host-cell reactivation of UV-irradiated and chemically-treated herpes simplex virus-1 by xeroderma pigmentosum, xp heterozygotes and normal skin fibroblasts  

International Nuclear Information System (INIS)

The host-cell reactivation of UV-irradiated and N-acetoxy-2-acetylamino-fluorene-treated herpes simplex virus type 1 strain MP was studied in normal and xeroderma pigmentosum human skin fibroblasts. Virus treated with either agent demonstrated lower survival in XP cells from complementation groups A, B, C and D than in normal fibroblasts. The relative reactivation ability of XP cells from the different genetic complementation groups was found to be the same for both irradiated and chemically treated virus. In addition, the inactivation kinetics for virus treated with either agent in the XP variant were comparable to that seen in normal skin fibroblasts. The addition of 2 or 4 mmoles caffeine to the post-infection assay medium had no effect on the inactivation kinetics of virus treated by either agent in the XP variant or in XP cells from the different genetic complementation groups. Treatment of the virus with nitrogen mustard resulted in equivalent survival in normal and XP genetic complementation group D cells. No apparent defect was observed in the ability of XP heterozygous skin fibroblasts to repair virus damaged with up to 100 ?g N-acetoxy-2-acetylaminofluorene per ml. These findings indicate that the repair of UV-irradiated and N-acetoxy-2-acetylaminofluorene-treated virus is accomplished by the same pathway or different pathways sharing a common intermediate step and that the excision defect of XP cells plays little if any role in the reactivation of nitrogen any role in the reactivation of nitrogen mustard treated virus. (Auth.)

174

Skin graft rejection in mice repopulated with marrow of the skin donor type: a Skn gene in a congenic line  

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Genetically anemic W/Wv mice and lethally irradiated wild-type mice were cured and populated by grafted marrow cells from donor mice of three congenic lines that differed at non-H-2 histocompatibility loci. Tail skin from mice of the same congenic lines was grafted 3-4 weeks later. In two cases, the recipients behaved as expected, no longer rejecting skin syngeneic with the marrow graft that had repopulated them. However, B6-H-24c skin was rejected by WBB6F1-W/Wv mice that were cured with B6-H-24c marrow showing a mean survival time of 9.9 weeks. It was rejected somewhat faster, with a mean survival time of 5.9 weeks, by W/Wv mice cured with marrow from other types of donors. Results were more variable in lethally irradiated WBB6F1-+/+ recipients of B6-H-24c marrow, but they also rejected B6-H-24c skin. Both types of recipients remained chimeras after the skin was rejected, showing more than 90% of the B6-H-24c hemoglobin type. This is the first report of a Skn gene in a congenic line

175

CopA3 peptide prevents ultraviolet-induced inhibition of type-I procollagen and induction of matrix metalloproteinase-1 in human skin fibroblasts.  

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Ultraviolet (UV) exposure is well-known to induce premature aging, which is mediated by matrix metalloproteinase-1 (MMP-1) activity. A 9-mer peptide, CopA3 (CopA3) was synthesized from a natural peptide, coprisin, which is isolated from the dung beetle Copris tripartitus. As part of our continuing search for novel bioactive natural products, CopA3 was investigated for its in vitro anti-skin photoaging activity. UV-induced inhibition of type-I procollagen and induction of MMP-1 were partially prevented in human skin fibroblasts by CopA3 peptide in a dose-dependent manner. At a concentration of 25 ?M, CopA3 nearly completely inhibited MMP-1 expression. These results suggest that CopA3, an insect peptide, is a potential candidate for the prevention and treatment of skin aging. PMID:24853614

Kim, Dong-Hee; Kim, Han-Hyuk; Kim, Hyeon-Jeong; Jung, Hyun-Gug; Yu, Jae-Myo; Lee, Eun-Su; Cho, Yong-Hun; Kim, Dong-In; An, Bong-Jeun

2014-01-01

176

CopA3 Peptide Prevents Ultraviolet-Induced Inhibition of Type-I Procollagen and Induction of Matrix Metalloproteinase-1 in Human Skin Fibroblasts  

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Full Text Available Ultraviolet (UV exposure is well-known to induce premature aging, which is mediated by matrix metalloproteinase-1 (MMP-1 activity. A 9-mer peptide, CopA3 (CopA3 was synthesized from a natural peptide, coprisin, which is isolated from the dung beetle Copris tripartitus. As part of our continuing search for novel bioactive natural products, CopA3 was investigated for its in vitro anti-skin photoaging activity. UV-induced inhibition of type-I procollagen and induction of MMP-1 were partially prevented in human skin fibroblasts by CopA3 peptide in a dose-dependent manner. At a concentration of 25 ?M, CopA3 nearly completely inhibited MMP-1 expression. These results suggest that CopA3, an insect peptide, is a potential candidate for the prevention and treatment of skin aging.

Dong-Hee Kim

2014-05-01

177

Formation and repair of DNA damage induced by indirect action of ultraviolet light in normal and xeroderma pigmentosum skin fibroblasts  

International Nuclear Information System (INIS)

The definitions of direct and indirect DNA damage are as follows: direct action, ''the attack of DNA occurs by a primary agent or a chemical derivative of the primary agent'', and indirect action, ''the attack of DNA occurs by active-O2 species which are formed by the reaction of a primary agent with a non-DNA target''. Comparative studies were performed on the formation and repair of DNA lesions induced by monochromatic light at 254, 265 and 313 nm. Attention was focused on human cells in culture, in particular on the skin fibroblasts from normal individuals and the patients with the autosomal recessive disease, Xerodermal pigmentosum (XP). The DNA lesions induced by the indirect action of active O2 species (superoxide-radicals, OH-radicals, singlet oxygen) become important. The most remarkable observation after the 313 nm irradiation on XP strains at 37 deg is the increased immediate fragmentation of pre-existing, parental DNA in the XP-variant strains. A late step in excision repair such as the ligation of parental DNA fragments may be deficient in the XP variants. The nuclease function involved in the removal of radiation lesions could be more active in the XP variants, or glycosylase function could be more active. The abnormality in de novo DNA synthesis in the XP variants may be the reflection of the primary defects in the repair of parental DNA templates rather than the defects in ''post-replication repair''. The abnormal function in DNA mn repair''. The abnormal function in DNA metabolism in the XP variants affects in parallel the repair of parental DNA and de novo DNA synthesis. (Yamashita, S.)

178

Altered transcriptome signature of phenotypically normal skin fibroblasts heterozygous for CDKN2A in familial melanoma: relevance to early intervention.  

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Familial melanoma (FM) is a dominantly heritable cancer that is associated with mutations in the tumor suppressor CDKN2A/p16. In FM, a single inherited "hit" occurs in every somatic cell, enabling interrogation of cultured normal skin fibroblasts (SFs) from FM gene carriers as surrogates for the cell of tumor origin, namely the melanocyte. We compared the gene expression profile of SFs from FM individuals with two distinct CDKN2A/p16 mutations (V126D-p16 and R87P-p16) with the gene expression profile of SFs from age-matched individuals without p16 mutations and with no family history of melanoma. We show an altered transcriptome signature in normal SFs bearing a single-hit inherited mutation in the CDKN2A/p16 gene, wherein some of these abnormal alterations recapitulate changes observed in the corresponding cancer. Significantly, the extent of the alterations is mutation-site specific with the R87P-p16 mutation being more disruptive than the V126D-p16 mutation. We also examined changes in gene expression after exposure to ultraviolet (UV) radiation to define potential early biomarkers triggered by sun exposure. UV treatment of SFs from FM families induces distinct alterations in genes related to cell cycle regulation and DNA damage responses that are also reported to be dysregulated in melanoma. Importantly, these changes were diametrically opposed to UV-induced changes in SF from normal controls. We posit that changes identified in the transcriptome of SF from FM mutation carriers represent early events critical for melanoma development. As such, they may serve as specific biomarkers of increased risk as well as molecular targets for personalized prevention strategies in high-risk populations. PMID:23371019

Fan, Meiyun; Pfeffer, Susan R; Lynch, Henry T; Cassidy, Pamela; Leachman, Sancy; Pfeffer, Lawrence M; Kopelovich, Levy

2013-01-01

179

Differential translocation of heat shock factor-1 after mild and severe stress to human skin fibroblasts undergoing aging in vitro.  

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Repeated exposure to mild heat shock (HS) has been shown to induce a wide range of health promoting hormetic effects in various biological systems, including human cells undergoing aging in vitro. In order to understand how cells distinguish between mild and severe stress, we have investigated the extent of early and immediate HS response by analyzing the nuclear translocation of the transcription factor heat shock factor-1 (HSF1), in serially passaged normal adult human facial skin fibroblasts exposed to mild (41 °C) or severe (43 °C) HS. Cells respond differently when exposed to mild and severe HS at different passage levels in terms of the extent of HSF1 translocation. In early passage young cells there was a 5-fold difference between mild and severe HS in the extent of HSF1 translocation. However, in near senescent late passage cells, the difference between mild and severe stress in terms of the extent of HSF1 translocation was reduced to less than 2-fold. One of the reasons for this age-related attenuation of heat shock response is due to the fact there was a higher basal level of HSF1 in the nuclei of late passage cells, which is indicative of increased intrinsic stress during cellular aging. These observations are consistent with previously reported data that whereas repeated mild stress given at younger ages can slow down aging and increase the lifespan, the same level of stress given at older ages may not provide the same benefits. Therefore, elucidating the early and immediate steps in the induction of stress response can be useful in deciding whether a particular level of stress is potentially hormetically beneficial or not. PMID:25193128

Demirovic, Dino; de Toda, Irene Martinez; Nizard, Carine; Rattan, Suresh I S

2014-12-01

180

Differential translocation of heat shock factor-1 after mild and severe stress to human skin fibroblasts undergoing aging in vitro  

DEFF Research Database (Denmark)

Repeated exposure to mild heat shock (HS) has been shown to induce a wide range of health promoting hormetic effects in various biological systems, including human cells undergoing aging in vitro. In order to understand how cells distinguish between mild and severe stress, we have investigated the extent of early and immediate HS response by analyzing the nuclear translocation of the transcription factor heat shock factor-1 (HSF1), in serially passaged normal adult human facial skin fibroblasts exposed to mild (41°C) or severe (43°C) HS. Cells respond differently when exposed to mild and severe HS at different passage levels in terms of the extent of HSF1 translocation. In early passage young cells there was a 5-fold difference between mild and severe HS in the extent of HSF1 translocation. However, in near senescent late passage cells, the difference between mild and severe stress in terms of the extent of HSF1 translocation was reduced to less than 2-fold. One of the reasons for this age-related attenuationof heat shock response is due to the fact there was a higher basal level of HSF1 in the nuclei of late passage cells, which is indicative of increased intrinsic stress during cellular aging. These observations are consistent with previously reported data that whereas repeated mild stress given at younger ages can slow down aging and increase the lifespan, the same level of stress given at older ages may not provide the same benefits. Therefore, elucidating the early and immediate steps in the induction of stress response can be useful in deciding whether a particular level of stress is potentially hormetically beneficial or not.

Demirovic, Dino; de Toda, Irene Martinez

2014-01-01

181

Mécano-Stimulation™ of the skin improves sagging score and induces beneficial functional modification of the fibroblasts: clinical, biological, and histological evaluations  

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Background Loss of mechanical tension appears to be the major factor underlying decreased collagen synthesis in aged skin. Numerous in vitro studies have shown the impact of mechanical forces on fibroblasts through mechanotransduction, which consists of the conversion of mechanical signals to biochemical responses. Such responses are characterized by the modulation of gene expression coding not only for extracellular matrix components (collagens, elastin, etc.) but also for degradation enzymes (matrix metalloproteinases [MMPs]) and their inhibitors (tissue inhibitors of metalloproteinases [TIMPs]). A new device providing a mechanical stimulation of the cutaneous and subcutaneous tissue has been used in a simple, blinded, controlled, and randomized study. Materials and methods Thirty subjects (aged between 35 years and 50 years), with clinical signs of skin sagging, were randomly assigned to have a treatment on hemiface. After a total of 24 sessions with Mécano-Stimulation™, biopsies were performed on the treated side and control area for in vitro analysis (dosage of hyaluronic acid, elastin, type I collagen, MMP9; equivalent dermis retraction; GlaSbox®; n=10) and electron microscopy (n=10). Furthermore, before and after the treatment, clinical evaluations and self-assessment questionnaire were done. Results In vitro analysis showed increases in hyaluronic acid, elastin, type I collagen, and MMP9 content along with an improvement of the migratory capacity of the fibroblasts on the treated side. Electron microscopy evaluations showed a clear dermal remodeling in relation with the activation of fibroblast activity. A significant improvement of different clinical signs associated with skin aging and the satisfaction of the subjects were observed, correlated with an improvement of the sagging cheek. Conclusion Mécano-Stimulation is a noninvasive and safe technique delivered by flaps microbeats at various frequencies, which can significantly improve the skin trophicity. Results observed with objective measurements, ie, in vitro assessments and electron microscopy, confirm the firming and restructuring effect clinically observed. PMID:25673979

Humbert, Philippe; Fanian, Ferial; Lihoreau, Thomas; Jeudy, Adeline; Elkhyat, Ahmed; Robin, Sophie; Courderot-Masuyer, Carol; Tauzin, Hélène; Lafforgue, Christine; Haftek, Marek

2015-01-01

182

EFFECTS OF CIPROFLOXACIN ON GROWTH OF HUMAN RHABDOMYOSARCOMA (RD AND RAT EMBRYO FIBROBLASTS (REF CELL LINES: IN VITRO STUDY  

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Full Text Available The possible anti-proliferative effects of ciprofloxacin utilizing cell lines obtained from different sources [human rhabdomyosarcoma (RD and transitional rat embryo fibroblasts (REF – (passage89] were studied. The present study was carried out from January 2011 to May 2011.Each of cell lines mentioned above was exposed to various concentrations of ciprofloxacin at concentrations (from 62.5 to 1000 mcg/ml for 48hours in addition to control (zero concentration. Four replicates were used for each data point and the results were expressed as mean ± SD. Ciprofloxacin caused significant growth inhibition on both cell lines only at 1000 mcg/ml concentration; but does not exert in vitro inhibitory effect on either human rhabdomyosarcoma (RD or transformed rat embryo fibroblasts (REF when assayed at concentrations of less than 1000 micrograms/ml.

Nada N. Al-Shawi

2012-12-01

183

Absence of correlations between the radiosensitivity of human T-lymphocytes at G0 and skin fibroblasts at log phase from the same individuals  

International Nuclear Information System (INIS)

Matched samples of peripheral T-lymphocytes and skin fibroblasts from a total of 22 patients who underwent various surgical procedures were tested for a dose-survival study using loss of colony-forming ability as the end point. The results showed that the mean D10 (the dose required to kill 90 % of the cells) ±SD was 3.58 ± 0.21 Gy for T-lymphocytes irradiated at G0 and 3.19 ± 0.37 Gy for skin fibroblasts irradiated at log phase. The coefficient of variation was found to be 6 % and 11 %, respectively. Contrary to expectation, regression analysis of the D10 values for the two cell types revealed no significant correlations. The absence of correlation is most probably derived from the fact that the apparent interindividual variability of dose-survival curves is largely caused by random experimental fluctuations, at least for lymphocytes. Possible reasons for the greater variability observed in the fibroblast assay are discussed. (author)

184

The treatment effects of cultured epidermis, basic fibroblast growth factor and the combination of these two treatments in a radiation skin ulcer model (rat)  

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The objective of this study was to evaluate the treatment effects of cultured epidermis, basic fibroblast growth factor (b-FGF) and the combination of these two treatments in a radiation skin ulcer model. The subjects were 9-week-old male inbred line rats and divided into two parts. Rats in one part were applied X-ray and rats in the other part were not. The dose of X-ray was 20 Gy. Wounds were full-thickness wounds. The ways of treatment were divided into four groups: control group, cultured epidermis group, b-FGF group, combination group (cultured epidermis+b-FGF). Wounds were observed on 5, 8, 11, 14, 17, 20, 23, 26 days after treatment. Wound healing rate was calculated and days needed to heal were counted. Relative hardness of scars was measured on the day of epithelization and on 12 and 21 days after epithelization. Wounds applied X-ray: Mean wound healing rate of cultured epidermis group and combination group was significantly higher than that of the two other groups on 8 and 11 days after treatment. Mean relative hardness of scars of cultured epidermis group and combination group was significantly lower than that of the two other groups on all measurement days. Mean days needed to heal of cultured epidermis group were significantly shorter than those of control group and b-FGF group. And those of combination group were significantly shorter than those of b-FGF group. As the shorter the days from making scars became, relative hardness of scars got lower. Woundslative hardness of scars got lower. Wounds without X-ray: Mean wound healing rate of combination group was significantly lower than that of cultured epidermis group and control group on 5 days after treatment. Cultured epidermis graft can be an effective treatment for radiation skin ulcer. b-FGF can weaken the treatment effect of cultured epidermis graft depending on its density. There can be a positive correlation between relative hardness of scars and the days from making scars. (author)

185

Genome-wide differential gene expression in immortalized DF-1 chicken embryo fibroblast cell line  

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Full Text Available Abstract Background When compared to primary chicken embryo fibroblast (CEF cells, the immortal DF-1 CEF line exhibits enhanced growth rates and susceptibility to oxidative stress. Although genes responsible for cell cycle regulation and antioxidant functions have been identified, the genome-wide transcription profile of immortal DF-1 CEF cells has not been previously reported. Global gene expression in primary CEF and DF-1 cells was performed using a 4X44K chicken oligo microarray. Results A total of 3876 differentially expressed genes were identified with a 2 fold level cutoff that included 1706 up-regulated and 2170 down-regulated genes in DF-1 cells. Network and functional analyses using Ingenuity Pathways Analysis (IPA, Ingenuity® Systems, http://www.ingenuity.com revealed that 902 of 3876 differentially expressed genes were classified into a number of functional groups including cellular growth and proliferation, cell cycle, cellular movement, cancer, genetic disorders, and cell death. Also, the top 5 gene networks with intermolecular connections were identified. Bioinformatic analyses suggested that DF-1 cells were characterized by enhanced molecular mechanisms for cell cycle progression and proliferation, suppressing cell death pathways, altered cellular morphogenesis, and accelerated capacity for molecule transport. Key molecules for these functions include E2F1, BRCA1, SRC, CASP3, and the peroxidases. Conclusions The global gene expression profiles provide insight into the cellular mechanisms that regulate the unique characteristics observed in immortal DF-1 CEF cells.

Kong Byung-Whi

2011-11-01

186

Effects of macelignan isolated from Myristica fragrans (nutmeg) on expression of matrix metalloproteinase-1 and type I procollagen in UVB-irradiated human skin fibroblasts  

International Nuclear Information System (INIS)

Exposure to ultraviolet (UV) light causes premature skin aging that is associated with upregulated matrix metalloproteinases (MMPs) and decreased collagen synthesis. Macelignan, a natural lignan compound isolated from Myristica fragrans HOUTT. (nutmeg), has been reported to possess antioxidant and antiinflammatory activities. This study assessed the effects of macelignan on photoaging and investigated its mechanisms of action in UV-irradiated human skin fibroblasts (Hs68) by reverse transcription-polymerase chain reaction, Western blot analysis, 2', 7'-dichlorofluorescein diacetate assay, and enzyme-linked immunosorbent assay. Our results show that macelignan attenuated UV-induced MMP-1 expression by suppressing phosphorylation of mitogen-activated protein kinases (MAPKs) induced by reactive oxygen species. Macelignan also increased type I procollagen expression and secretion through transforming growth factor ? (TGF-?)/Smad signaling. These findings indicate that macelignan regulates the expression of MMP-1 and type I procollagen in UV-irradiated human skin fibroblasts by modulating MAPK and TGF-?/Smad signaling, suggesting its potential as an efficacious antiphotoaging agent. (author)

187

Effects of macelignan isolated from Myristica fragrans (Nutmeg) on expression of matrix metalloproteinase-1 and type I procollagen in UVB-irradiated human skin fibroblasts.  

Science.gov (United States)

Exposure to ultraviolet (UV) light causes premature skin aging that is associated with upregulated matrix metalloproteinases (MMPs) and decreased collagen synthesis. Macelignan, a natural lignan compound isolated from Myristica fragrans HOUTT. (nutmeg), has been reported to possess antioxidant and antiinflammatory activities. This study assessed the effects of macelignan on photoaging and investigated its mechanisms of action in UV-irradiated human skin fibroblasts (Hs68) by reverse transcription-polymerase chain reaction, Western blot analysis, 2',7'-dichlorofluorescein diacetate assay, and enzyme-linked immunosorbent assay. Our results show that macelignan attenuated UV-induced MMP-1 expression by suppressing phosphorylation of mitogen-activated protein kinases (MAPKs) induced by reactive oxygen species. Macelignan also increased type I procollagen expression and secretion through transforming growth factor ? (TGF-?)/Smad signaling. These findings indicate that macelignan regulates the expression of MMP-1 and type I procollagen in UV-irradiated human skin fibroblasts by modulating MAPK and TGF-?/Smad signaling, suggesting its potential as an efficacious antiphotoaging agent. PMID:23037157

Lee, Kyung-Eun; Mun, Sukyeong; Pyun, Hee-Bong; Kim, Myung-Suk; Hwang, Jae-Kwan

2012-01-01

188

Effects of licarin E on expression of matrix metalloproteinase-1 and type-1 procollagen in UVB-irradiated human skin fibroblasts.  

Science.gov (United States)

Ultraviolet B (UVB) radiation induces photoaging by upregulating the expression of matrix metalloproteinase (MMP) and decreasing collagen synthesis in human skin cells. This study evaluated the effects of licarin E isolated from mace, the aril of Myristica fragrans Houtt., on MMP-1 and type-1 procollagen levels in UVB-irradiated human skin fibroblasts. Powdered mace extracted with 95% ethanol was used and licarin E isolated by preparative high-performance liquid chromatography. In addition, western blot analysis, reverse transcription PCR and electrophoretic mobility shift assay were used to evaluate the effects of licarin E and its molecular mechanism. It was found that licarin E attenuated UVB-induced MMP-1 expression by inactivating mitogen-activated protein kinases (MAPKs), thereby inhibiting activator protein 1. Licarin E also increased type-1 procollagen expression by stimulating transforming growth factor ? (TGF?)/Smad signaling. The findings show that licarin E positively regulates the expression of MMP-1 and type-1 procollagen in UVB-irradiated human skin fibroblasts through MAPK and TGF? signaling, suggesting its potential as a potent antiphotoaging agent. PMID:21544886

Kwon, Yi-Young; Kim, Daeyoung; Kim, Jaekyung; Hwang, Jae-Kwan

2011-12-01

189

The Apoptotic Effects of the P300 Activator on Breast Cancer and Lung Fibroblast Cell Lines  

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Full Text Available Background: P300 is an enzyme that acetylates histones during stress. It alsoacetylates several non-histone proteins, including P53 which is the most important tumorsuppressor gene. P53 plays an important role in the apoptosis of tumor cells. Hereby,this study describes the potency of cholera toxin B subunit as a P300 activator to induceapoptosis in a breast cancer cell line (MCF-7 and a lung fibroblast cell line (MRC-5as a non-tumorigenic control sample. Methods: MCF-7 and MRC-5 were cultured in RPMI-1640 and treated with orwithout cholera toxin B subunit at the concentration of 85.43 ?mol/L, based on the half-maximal inhibitory concentration index at different times (24, 48 and 72 h. Thepercentage of apoptotic cells was measured by flow cytometry. Real-time quantitativeRT-PCR was performed to estimate the mRNA expression of P300 in MCF-7 and MRC-5 with cholera toxin B subunit at different times. We used the ELISA and Bradford proteintechniques to detect levels of total and acetylated P53 protein generated in MCF-7 andMRC-5. Results: Our findings indicated that the cholera toxin B subunit effectively andsignificantly induced more apoptosis in MCF-7 compared to MRC-5. We showed thatexpression of P300 up-regulated by increasing the time of the cholera toxin B subunittreatment in MCF-7 but not in MRC-5. In addition, the acetylated and total P53protein levels increased more in MCF-7 cells than in MRC-5 cells.Conclusion: Cholera toxin B subunit induced significant cell death in MCF-7, butit could be well tolerated in MRC-5. Therefore, cholera toxin B subunit can besuggested as an anti-cancer agent.

Mohammad Reza Salahshoor

2013-10-01

190

In vitro correction of ARSA deficiency in human skin fibroblasts from metachromatic leukodystrophy patients after treatment with microencapsulated recombinant cells.  

Science.gov (United States)

Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder due to arylsulfatase A (ARSA) deficiency that affects primarily the central nervous system. Ongoing treatments include enzyme replacement therapy and bone marrow transplantation, both limited in their effects due to the blood-brain barrier. An alternative approach would be the in situ implantation of encapsulated cells over expressing ARSA. Based on that, we tested the ability of encapsulated BHK cells over expressing ARSA to correct the enzyme deficiency in MLD patients' fibroblasts. Three groups were analyzed: fibroblasts treated with ARSA-over expressing BHK cells (rBHK) trapped in alginate capsules (capsules group), fibroblasts treated with supernatant of non-encapsulated rBHK (uptake control) and fibroblasts treated with empty capsules (empty group). Untreated and normal fibroblasts were used as controls. rBHK obtained by clone selection after non-viral transfection with pTARSA-CMV2. ARSA activity was measured after 1, 2, 3 and 4 weeks of treatment and beta-gal was used as reference enzyme. Statistical analysis was performed using ANOVA and Tukey's test. Normal fibroblasts showed ARSA activity of 23.9 + /- 2.01 nmol/h/mg of protein, whereas untreated MLD fibroblasts had the low ARSA activity (2.22 + /- 0.17). In the empty group, ARSA activity was equal to that of untreated fibroblasts (2.71 + /- 0.34). Capsules and uptake control groups showed higher enzymatic activity levels, compared to MLD untreated, 23.42 + /- 6.39 and 42.35 + /- 5.20, respectively (p < 0.01 for all groups). Encapsulated rBHK clones show potential as a new therapeutic strategy for the treatment of MLD, reaching normal enzyme levels in human MLD fibroblasts. PMID:18797988

Lagranha, Valeska Lizzi; Baldo, Guilherme; de Carvalho, Talita Giacomet; Burin, Maira; Saraiva-Pereira, Maria Luiza; Matte, Ursula; Giugliani, Roberto

2008-12-01

191

Low-dose radiation activates Nrf1/2 through reactive species and the Ca2+/ERK1/2 signaling pathway in human skin fibroblast cells  

OpenAIRE

In the current study, we explored the effect of LDR on theactivation of Nrfs transcription factor involved in cellular redoxevents. Experiments were carried out utilizing 0.05 and 0.5 GyX-ray irradiated normal human skin fibroblast HS27 cells. Theresults showed LDR induced Nrf1 and Nrf2 activation andexpression of antioxidant genes HO-1, Mn-SOD, and NQO1.In particular, 0.05 Gy-irradiation increased only Nrf1 activation,but 0.5 Gy induced both Nrf1 and Nrf2 activation.LDR-mediated Nrf1/2 activ...

Eun Kyeong Lee; Jin-Ah Kim; Seong Joon Park; Jeung Ki Kim; Kyu Heo; Kwang Mo Yang; Tae Gen Son

2013-01-01

192

Carrier detection in the testicular feminisation syndrome: deficient 5 alpha-dihydrotestosterone binding in cultured skin fibroblasts from the mothers of patients with complete androgen insensitivity.  

OpenAIRE

Specific binding of 5 alpha-dihydrotestosterone (androgen receptor activity) could not be detected in cultured genital skin fibroblasts (GSF) from two patients with complete androgen insensitivity (CAI). In GSF from the mother of one patient, androgen receptor activity (8.5 fmol/mg cell protein) was reduced in comparison with controls (34.0 +/- 10.1 (SD) fmol/mg protein n = 15). These results favour X linked inheritance of CAI and X inactivation at the androgen receptor locus. Androgen recept...

Hodgins, M. B.; Duke, E. M.; Ring, D.

1984-01-01

193

Screening of medicinal and edible plants in Okinawa, Japan, for enhanced proliferative and collagen synthesis activities in NB1RGB human skin fibroblast cells.  

Science.gov (United States)

To identify plants with bioactive potential for skin care, methanol extracts of 56 plant parts from 47 medical and edible plants cultivated in Okinawa were tested for their proliferative effects on NB1RGB skin fibroblast cells. Extracts from six plants, Bischofia javanica, Colocasia esculenta, Melaleuca alternifolia, Piper angustifolia, Jasminum sambac, and Curcuma longa, showed higher NB1RGB cell proliferation activity (>10%) than the control, at various concentrations. Among the six extracts, only the C. longa extract caused an increase in collagen synthesis in NB1RGB cells, as compared to treatment with the positive control, ascorbic acid (AsA). Expression of the collagen synthesis marker, transforming growth factor-?1, was higher after treatment with the C. longa extract than with AsA. PMID:23221723

Takahashi, Makoto; Asikin, Yonathan; Takara, Kensaku; Wada, Koji

2012-01-01

194

A role for Nrf2 in UVA-mediated heme oxygenase induction and protection from membrane damage in human skin fibroblasts  

Science.gov (United States)

Our previous study has shown that Ultraviolet-A (UVA) irradiation induces heme oxygenase 1 (HO-1) expression in cultured human primary skin fibroblasts FEK4. In the present study, we demonstrate a coordinated induction of HO-1 and NF-E2-related factor 2 (Nrf2) following UVA irradiation or hemin treatment. The induction of HO-1 by either UVA irradiation or hemin treatment was largely abolished by down-regulation of Nrf2 with its targeted short interfering RNA (siNrf2). The study further reveals that knockdown of Nrf2 protein increased UVA-induced cell death measured by MTS assay. These findings together indicate that Nrf2-mediated induction of HO-1 expression may provide a cytoprotection for human skin cells from oxidative damage.

Li, Haibin; Li, Linhao; Deng, Linhong; Singh, Gurinder; Tyrrell, Rex M.; Zhong, J. Li

2010-11-01

195

Characterization of fibroblast growth factor receptor 2 overexpression in the human breast cancer cell line SUM-52PE  

OpenAIRE

The fibroblast growth factor receptor (FGFR)2 gene has been shown to be amplified in 5-10% of breast cancer patients. A breast cancer cell line developed in our laboratory, SUM-52PE, was shown to have a 12-fold amplification of the FGFR2 gene, and FGFR2 message was found to be overexpressed 40-fold in SUM-52PE cells as compared with normal human mammary epithelial (HME) cells. Both human breast cancer (HBC) cell lines and HME cells expressed two FGFR2...

Tannheimer, Stacey L.; Rehemtulla, Alnawaz; Ethier, Stephen P.

2000-01-01

196

MRI Breast Skin-line Segmentation and Removal using Integration Method of Level Set Active Contour and Morphological Thinning Algorithms  

Directory of Open Access Journals (Sweden)

Full Text Available In the MRI breast Computer Aided Detection systems, skin-line segmentation and removal is a vital process. Similar intensity levels of the skin-line and the other parts of the breast image such as; dense tissue and tumor could possibly lead to faulty tumor segmentation if the skin-line is not removed correctly. In this study, a technique for skin-line segmentation and removal is presented. The approach integrates two algorithms. Level Set Active Contour algorithm is used to segment the breast skin border; the Morphological Thinning Algorithm is used to delete the detected breast skin-line. The approach is applied and tested on the RIDER breast MRI dataset and the results are evaluated using six measures. The evaluation results show high performance for the proposed approach with accuracy of 0.9607 for the skin-line segmentation stage and 0.9099 for the removal stage.

Nor Ashidi Mat Isa

2012-01-01

197

Induction of cAMP-dependent protein kinase A activity in human skin fibroblasts and rat osteoblasts by extremely low-frequency electromagnetic fields  

Energy Technology Data Exchange (ETDEWEB)

Sinusoidal extremely low-frequency electromagnetic fields (ELF-EMF; 7-8 mT, 20 Hz) have already been shown to inhibit proliferation and to accelerate terminal differentiation of human skin fibroblasts in vitro. In order to elucidate the underlying processes of signal transduction, we analysed the activity of cAMP-dependent protein kinase (PKA). EMF exposure for 60 min resulted in an increased PKA activity in human skin fibroblasts (2-fold) and rat embryonic osteoblasts (1.7-fold). Long-term exposure for up to 7 days with a constant 1 h-on/1 h-off EMF exposure rhythm indicated a transient stimulation of PKA activity during the first two exposure rhythms followed by a decrease to the baseline levels of sham-exposed controls. Based on these results, we postulate that a modulation of proliferation and differentiation processes in cells of mesenchymal origin is triggered by an immediate and transient EMF-induced increase in PKA activity. (orig.)

Thumm, S.; Glock, S.; Haemmerle, H. [Natural and Medical Sciences Institute Reutlingen, University of Tuebingen (NMI), Markwiesenstrasse 55, D-72770 Reutlingen (Germany); Loeschinger, M.; Rodemann, H.P. [Section of Radiobiology and Molecular Environmental Research, University of Tuebingen, Roentgenweg 11, D-72076 Tuebingen (Germany)

1999-09-01

198

Protective effects of Chlorella-derived peptide on UVB-induced production of MMP-1 and degradation of procollagen genes in human skin fibroblasts.  

Science.gov (United States)

UV exposure is known to induce premature aging, which is mediated by matrix metalloproteinase-1 (MMP-1) activity. MMP-1 mRNA expression is up-regulated by elevated cysteine-rich 61 (CYR61) and monocyte chemoattractant protein-1 (MCP-1) via action of transcription factor AP-1. Collagen is degraded by MMP-1 activity but synthesized by transforming growth factor-? (TGF-?) signal. Chlorella has been shown to inhibit UVB-induced MMP-1 level, however its regulatory molecular mechanisms have not been studied. In this study, Chlorella derived peptide (CDP) was added to skin fibroblasts after UVB irradiation and the expression of MMP-1, CYR61, procollagen, c-fos, c-jun, and TGF-? receptor (TbRII) mRNA and MCP-1 production were investigated. CDP (10 or 5mg/ml) diminished UVB-induced MMP-1 and CYR61 mRNA expression and MCP-1 production, whereas, UVB-suppressed procollagen and TbRII mRNA was restored by CDP treatment. UVB-induced c-fos and c-jun expressions were also inhibited by the CDP treatment. Taken together, CDP inhibits UVB-induced MMP-1 expression in skin fibroblasts by suppressing expression of AP-1 and CYR61 and MCP-1 production. PMID:21397653

Chen, Chiu-Lan; Liou, Shu-Fen; Chen, Su-Jong; Shih, Mei-Fen

2011-06-01

199

Establishment and characterization of fetal fibroblast cell lines for generating human lysozyme transgenic goats by somatic cell nuclear transfer.  

Science.gov (United States)

This study was performed to qualify goat fetal fibroblast (GFF) cell lines for genetic modification and somatic cell nuclear transfer (SCNT) to produce human lysozyme (hLYZ) transgenic goats. Nine GFF cell lines were established from different fetuses, and the proliferative lifespan and chromosomal stability were analyzed. The results suggested that cell lines with a longer lifespan had stable chromosomes compared with those of cells lines with a shorter lifespan. According to the proliferative lifespan, we divided GFF cell lines into two groups: cell lines with a long lifespan (GFF1/2/7/8/9; group L) and cell lines with a short lifespan (GFF3/4/5/6; group S). Next, a hLYZ expression vector was introduced into these cell lines by electroporation. The efficiencies of colony formation, expansion in culture, and the quality of transgenic clonal cell lines were significant higher in group L than those in group S. The mean fusion rate and blastocyst rate in group L were higher than those in group S (80.3 ± 1.7 vs. 65.1 ± 4.2 % and 19.5 ± 0.6 vs. 15.1 ± 1.1 %, respectively, P cell lines has a major effect on the efficiency to produce transgenic cloned goats. Therefore, the proliferative lifespan of primary cells may be used as a criterion to characterize the quality of cell lines for genetic modification and SCNT. PMID:23335060

Liu, Jun; Luo, Yan; Zheng, Liming; Liu, Qingqing; Yang, Zhongcai; Wang, Yongsheng; Su, Jianmin; Quan, Fusheng; Zhang, Yong

2013-10-01

200

Quantitative proteomic analysis revealed N'-nitrosonornicotine-induced down-regulation of nonmuscle myosin II and reduced cell migration in cultured human skin fibroblast cells.  

Science.gov (United States)

The association of tobacco smoke with decreased cell motility and wound healing is well documented; however, the cellular mechanisms and specific toxic tobacco constituents responsible for this effect are not well understood. Tobacco-specific N-nitrosamines (TSNAs) are among the most important classes of carcinogens found in tobacco products. The TSNA N'-nitrosonornicotine (NNN) is present at relatively high levels in tobacco and its smoke, as well as second- and third-hand smoke. To investigate the cellular pathways that are perturbed upon NNN exposure, we employed a quantitative proteomic approach, utilizing stable isotope labeling by amino acids in cell culture and mass spectrometry, to assess the NNN-induced alteration of protein expression in GM00637 human skin fibroblast cells. With this approach, we were able to quantify 2599 proteins, 191 of which displayed significantly changed expression following NNN exposure. One of the main findings from our proteomic analysis was the down-regulation of six different subunits of myosin, particularly nonmuscle myosin II heavy chain, isoforms A, B, and C. In addition, we found the altered expression of several extracellular matrix proteins and proteins involved in cellular adhesion. Together, our quantitative proteomic results suggested that NNN exposure may interfere with fibroblast motility. An in vitro scratch wound assay result supported that NNN exposure reduced the ability of dermal fibroblast to migrate into the scratched area. The results from the present study offer novel insights into the cellular mechanisms of NNN toxicity and identify NNN as a specific tobacco constituent that contributes to decreased fibroblast migration. PMID:23305604

Prins, John M; Wang, Yinsheng

2013-03-01

201

Quantitative Proteomic Analysis Revealed N?-Nitrosonornicotine-induced Down-regulation of Non-muscle Myosin II and Reduced Cell Migration in Cultured Human Skin Fibroblast Cells  

Science.gov (United States)

The association of tobacco smoke with decreased cell motility and wound healing is well documented; however, the cellular mechanisms and specific toxic tobacco constituents responsible for this effect are not well understood. Tobacco-specific N-nitrosamines (TSNAs) are among the most important classes of carcinogens found in tobacco products. The TSNA N?-nitrosonornicotine (NNN) is present at relatively high levels in tobacco and its smoke, as well as second- and third-hand smoke. To investigate the cellular pathways that are perturbed upon NNN exposure, we employed a quantitative proteomic approach, utilizing stable isotope labeling by amino acids in cell culture and mass spectrometry, to assess the NNN-induced alteration of protein expression in GM00637 human skin fibroblast cells. With this approach, we were able to quantify 2599 proteins, 191 of which displayed significantly changed expression following NNN exposure. One of the main findings from our proteomic analysis was the down-regulation of six different subunits of myosin, particularly non-muscle myosin II heavy chain, isoforms A, B, and C. In addition, we found the altered expression of several extracellular matrix proteins and proteins involved in cellular adhesion. Together, our quantitative proteomic results suggested that NNN exposure may interfere with fibroblast motility. An in vitro scratch wound assay results supported that NNN exposure reduced the ability of dermal fibroblast to migrate into the scratched area. The results from the present study offered novel insights into the cellular mechanisms of NNN toxicity and identified NNN as a specific tobacco constituent that contributes to decreased fibroblast migration. PMID:23305604

Prins, John M.; Wang, Yinsheng

2013-01-01

202

Evidence for a positive correlation between in vitro radiosensitivity of normal human skin fibroblasts and the occurrence of subcutaneous fibrosis after radiotherapy  

DEFF Research Database (Denmark)

A colony-forming assay of human skin fibroblast radiosensitivity was established in our laboratory and applied to primary skin biopsies from 12 women belonging to an unselected group of patients who received postmastectomy radiotherapy 10-12 years prior to this study. The aim was to investigate the relationship between in vitro radiosensitivity and the occurrence of subcutaneous fibrosis after radiotherapy. Early generations of normal skin fibroblasts in exponential growth were irradiated at room temperature at a high dose-rate to estimate the surviving fraction of colony-forming cells after single doses ranging from 1 to 8 Gy. A linear-quadratic cell survival curve was fitted to the data and from these fits the surviving fraction at 3.5 Gy (SF3.5) was estimated. Replicate experiments of different cell generations were made to validate the assay, and the between-patients variability was significantly larger than the assay variability for both SF2(p = 0.002) and SF3.5 (p = 0.04). Patients were treated in the period 1978-1982 with a dose per fraction between 2.7 and 3.9 Gy, a total of 12 fractions at two fractions per week. They were evaluated with respect to the occurrence of marked subcutaneous fibrosis in a total of 36 independent treatment fields. In each treatment field the total dose and dose per fraction at the relevant dosimetric reference depth as well as the length of follow-up were recorded. A previously derived LQ mixture model was applied to these data in order to determine the probability of marked fibrosis in that particular field. From this probability and the actually observed fibrosis, the excess risk of fibrosis was calculated, and this was averaged over the three treatment fields to obtain a single measure of clinical radiosensitivity. Increasing values of SF3.5 were statistically significantly correlated with decreasing probabilities of developing subcutaneous fibrosis (p = 0.014, Spearman's rank correlation test). Thus, this pilot study has demonstrated a positive correlation between in vivo radiosensitivity and normal skin fibroblasts and the clinical expression of subcutaneous fibrosis.

Johansen, J; Bentzen, SØren M

1994-01-01

203

A high molecular arabinogalactan from Ribes nigrum L.: influence on cell physiology of human skin fibroblasts and keratinocytes and internalization into cells via endosomal transport.  

Science.gov (United States)

An arabinogalactan protein (F2) was isolated in 1.5% yield from the seeds of Ribes nigrum L. (Grossulariaceae) by aqueous extraction and a one-step anion exchange chromatography on DEAE-Sephacel with 24% galactose, 43% arabinose, and 20% xylose as main carbohydrate residues. Methylation analysis revealed the presence of a 1,3-/1,3,6-galactose backbone, side chains from arabinose in different linkages, and terminal xylose residues. The polysaccharide which turned out to be an arabinogalactan protein had a molecular weight of >10(6) Da and deaggregated under chaotropic conditions. The cellular dehydrogenase activities (MTT and WST-1 tests) of human skin cells (fibroblasts, keratinocytes) as well as the proliferation rate of keratinocytes (BrdU incorporation ELISA) were significantly stimulated by the polymer at 10 and 100 microg/mL. F2 had no influence on differentiation status of keratinocytes and did not exhibit any cytotoxic potential (LDH test). The biological activity of F2 was not dependent on the high molecular weight. Influence of the polysaccharide on the gene expression of specific growth factors, growth factor receptors, signal proteins and marker proteins for skin cell proliferation, and differentiation by RT-PCR could not be shown. Gene array investigations indicated an increased expression of various genes encoding for catabolic enzymes, DNA repair, extracellular matrix proteins, and signal transduction factors. Removal of terminal arabinose residues by alpha-L-arabinofuranosidase did not influence the activity toward skin cells, while the treatment with beta-D-galactosidase yielded an inactive polysaccharide. The FITC-labeled polysaccharide was incorporated in a time-dependent manner into human fibroblasts (laser scanning microscopy) via endosomal transport. This internalization of the polysaccharide was inhibited by Cytochalasin B. PMID:19368904

Zippel, Janina; Deters, Alexandra; Pappai, Dirk; Hensel, Andreas

2009-05-26

204

Establishment of immortal normal and ataxia telangiectasia fibroblast cell lines by introduction of the hTERT gene  

International Nuclear Information System (INIS)

To establish immortal human cells, we introduced the human catalytic subunit of telomerase (hTERT) gene into skin fibroblast cells obtained from normal and ataxia telangiectasia (AT) individuals of Japanese origin. After hTERT introduction, these cells continue to grow beyond a population doubling number of 200 while maintaining their original radiosensitivity. Inductions of p53, phosphorylation of Serl5 in p53, and induction of p21 by X-ray irradiation in immortal cells derived from normal individual were not affected by the hTERT introduction. Both normal and AT immortal cells exhibited an apparent inhibition of growth as original primary cells when they reached confluence. Karyotype analysis has revealed that they are in a diploid range. These results suggest that cells immortalized by hTERT introduction retain their original characteristics except for immortalization, and that they may be useful for analyzing various effects of radiation on human cells. (author)

205

Establishment of immortal normal and ataxia telangiectasia fibroblast cell lines by introduction of the hTERT gene  

Energy Technology Data Exchange (ETDEWEB)

To establish immortal human cells, we introduced the human catalytic subunit of telomerase (hTERT) gene into skin fibroblast cells obtained from normal and ataxia telangiectasia (AT) individuals of Japanese origin. After hTERT introduction, these cells continue to grow beyond a population doubling number of 200 while maintaining their original radiosensitivity. Inductions of p53, phosphorylation of Serl5 in p53, and induction of p21 by X-ray irradiation in immortal cells derived from normal individual were not affected by the hTERT introduction. Both normal and AT immortal cells exhibited an apparent inhibition of growth as original primary cells when they reached confluence. Karyotype analysis has revealed that they are in a diploid range. These results suggest that cells immortalized by hTERT introduction retain their original characteristics except for immortalization, and that they may be useful for analyzing various effects of radiation on human cells. (author)

Nakamura, Hideaki; Fukami, Hiroko; Hayashi, Yuko; Kiyono, Tohru; Ishizaki, Kanji [Aichi Cancer Center, Nagoya (Japan). Research Inst.; Nakatsugawa, Shigekazu; Hamaguchi, Michinari [Nagoya Univ. (Japan). School of Medicine

2002-06-01

206

Intracellular insulin-receptor dissociation and segregation in a rat fibroblast cell line transfected with a human insulin receptor gene  

Energy Technology Data Exchange (ETDEWEB)

The cellular processing of insulin and insulin receptors was studied using a rat fibroblast cell line that had been transfected with a normal human insulin receptor gene, expressing approximately 500 times the normal number of native fibroblasts insulin receptors. These cells bind and internalize insulin normally. Biochemically assays based on the selective precipitation by polyethylene glycol of intact insulin-receptor complexes but not of free intracellular insulin were developed to study the time course of intracellular insulin-receptor dissociation. Fibroblasts were incubated with radiolabeled insulin at 4/sup 0/C, and internalization of insulin-receptor complexes was initiated by warming the cells to 37/sup 0/C. Within 2 min, 90% of the internalized radioactivity was composed of intact insulin-receptor complexes. The dissociation of insulin from internalized insulin-receptor complexes was markedly inhibited by monensin and chloroquine. Furthermore, chloroquine markedly increased the number of cross-linkable intracellular insulin-receptor complexes, as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis autoradiography. These findings suggest that acidification of intracellular vesicles is responsible for insulin-receptor dissociation. Physical segregation of dissociated intracellular insulin from its receptor was monitored. The results are consistent with the view that segregation of insulin and receptor occurs 5-10 min after initiation of dissociation. These studies demonstrate the intracellular itinerary of insulin-receptor complexes, including internalization, dissociation of insulin from the internalized receptor within an acidified compartment, segregation of insulin from the receptor, and subsequent ligand degradation.

Levy, J.R.; Olefsky, J.M.

1988-05-05

207

[Values of wettedness observed in clothed subject and the theoretical equal line of average skin temperature].  

Science.gov (United States)

In order to obtain the characteristics of the change of wettedness under constant average skin temperature 36 degrees C, the experiments were carried out using two young male subjects in a test chamber. From the basic measurements of both environmental parameters and human physiological responses, the authors found the wettedness value changeable even though the average skin temperature was constant. The following results were obtained under the conditions of resting-sitting, summer clothing worn, still air movement and constant average skin temperature 36 degrees C; 1. there is a positive correlation between wettedness and the environmental vapor pressure. 2. There is a negative correlation between wettedness and the air temperature. 3. There is a positive correlation between the evaporative heat loss from skin surface and the air temperature. 4. There is a negative correlation between the evaporative heat loss and the vapor pressure. 5. There is a negative correlation between wettedness and the evaporative heat loss. 6. Both maximum and minimum values of wettedness correspond to the average skin temperature. Considering the afore-mentioned results, the equal line of the average skin temperature does not form a straight line, but a curved line on the psychrometric chart. PMID:7916763

Mochida, T; Shimakura, K

1994-07-01

208

Establishment and genetic characteristics analysis of in vitro culture a fibroblast cell line derived from Wuzhishan miniature pig.  

Science.gov (United States)

Establishment of fibroblast cell lines of endangered pig breeds and research on the gene functions based on the cells made a significant contribution to the conservation and utilization of genetic resources. The Wuzhishan miniature pig ear marginal tissue fibroblast cell line (WPF22) from 22 samples, stocking 87 cryogenically-preserved vials, was successfully established by using primary explants technique and cell cryopreservation techniques. WPF22 cells were adherent, with a population doubling time of 30.2h. Chromosome karyotyping and G-banding analysis showed that >90.2% of cells were diploid (2n=38) prior to the 4th generation. Neither microbial contamination nor cross-contamination was detected by isoenzyme analyses. Cell viability was 97.8% before cryopreservation and 94.9% after recovery. To determine cell permeability, intracellular path and stability of exogenous proteins during the transduction, six fluorescent protein genes were transferred into fibroblasts by lipofectamine-mediated method. The transfection efficiency of six fluorescent protein genes fluctuated between 8.1% and 42.6%. ECFP and DsRed were mostly shown in cytoplasmic in dots around the nucleus, and EYFP and EGFP had a slightly stronger expression in the nucleus than in the cytoplasm, but without expression in some vacuoles. Every index of the WPF22 cell line meets all the standard quality controls of American type Culture Collection (ATCC). This research thus does not only preserve important genetic resources of Wuzhishan miniature pig at the cell level, but also serve as a valuable resource for genome, postgenome and somacloning research. PMID:24556363

Liu, Changqing; Guo, Yu; Lu, Taofeng; Li, Xiangchen; Guan, Weijun; Ma, Yuihui

2014-04-01

209

IN VITRO EFFECTS OF CEFTRIAXONE ON GROWTH OF HUMAN RHABDOMYOSARCOMA (RD AND ON RAT EMBRYO FIBROBLASTS (REF CELL LINES  

Directory of Open Access Journals (Sweden)

Full Text Available Many reports demonstrated that antibiotics like cefazoline, ciprofloxacin, trimethoprime-sulfamethoxazole and others have the ability to inhibit growth of various cell lines. Thus, this study was designed to investigate whether or not ceftriaxone may possess cell growth inhibition using In vitro study and utilizing two cell lines (human rhabdomyosarcoma (RD and rat embryo fibroblasts (REF. Various concentrations of Ceftriaxone (62.5, 125, 250, 500 and 1000 mcg/ml were utilized in this study. The drug relatively caused concentration-dependent inhibition on growth of the intended cell lines used in this study, where, the growth inhibition induced by the drug was statistically significant at 125mcg/ml and above. The results obtained from this work encourage further study of the possibility of clinical application of ceftriaxone to prevent the occurrence of different tumors.

Nada N. Al-Shawi

2013-02-01

210

Establishment and Characterization of Immortalized Gingival Epithelial and Fibroblastic Cell Lines for the Development of Organotypic Cultures.  

Science.gov (United States)

In vitro studies using 3D co-cultures of gingival cells can resemble their in vivo counterparts much better than 2D models that typically only utilize monolayer cultures with short-living primary cells. However, the use of 3D gingival models is still limited through lack of appropriate cell lines. We aimed to establish immortalized cell line models of primary human gingival epithelium keratinocytes (HGEK) and gingival fibroblasts (GFB). Immortalized cell lines (HGEK-16 and GFB-16) were induced by E6 and E7 oncoproteins of human papillomavirus. In addition, 3D multilayered organotypic cultures were formed by embedding GFB-16 cells within a collagen (Col) matrix and seeding of HGEK-16 cells on the upper surfaces. Cell growth was analyzed in both immortalized cell lines and their parental primary cells. The expression levels of cell type-specific markers, i.e. cytokeratin (CK) 10, CK13, CK16, CK18, CK19 for HGEK-16 and Col I and Col II for GFB-16, were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Expansion of the primary cultures was impeded at early passages, while the transformed immortalized cell lines could be expanded for more than 30 passages. In 3D cultures, immortalized HGEK formed a multilayer of epithelial cells. qRT-PCR showed that cell-specific marker expression in the 3D cultures was qualitatively and quantitatively closer to that in human gingival tissue than to monolayer cultures. These results indicate that immortalized gingival fibroblastic and epithelial cell lines can successfully form organotypic multilayered cultures and, therefore, may be useful tools for studying gingival tissue in vitro. © 2014 S. Karger AG, Basel. PMID:25471635

Bao, Kai; Akguel, Baki; Bostanci, Nagihan

2014-11-29

211

Spontaneous establishment of an Epstein-Barr virus-infected fibroblast line from the synovial tissue of a rheumatoid arthritis patient.  

OpenAIRE

An Epstein-Barr virus (EBV)-infected fibroblast line, designated DSEK, was spontaneously established from synovial tissue of a patient with rheumatoid arthritis (RA). DSEK cells expressed EBV nuclear antigens EBNA-1 and EBNA-2 and latent membrane protein LMP-1. Cell surface markers of DSEK cells were similar to those of EBV-negative fibroblast clones derived from synoviocytes and were negative for lymphocyte and macrophage markers. DSEK cells expressed CD44, CD58, and HLA-DR antigens and spon...

Koide, J.; Takada, K.; Sugiura, M.; Sekine, H.; Ito, T.; Saito, K.; Mori, S.; Takeuchi, T.; Uchida, S.; Abe, T.

1997-01-01

212

Differential Fibroblast Growth Factor 8 (FGF8)-Mediated Autoregulation of Its Cognate Receptors, Fgfr1 and Fgfr3, in Neuronal Cell Lines  

OpenAIRE

Fibroblast growth factors (FGFs) mediate a vast range of CNS developmental processes including neural induction, proliferation, migration, and cell survival. Despite the critical role of FGF signaling for normal CNS development, few reports describe the mechanisms that regulate FGF receptor gene expression in the brain. We tested whether FGF8 could autoregulate two of its cognate receptors, Fgfr1 and Fgfr3, in three murine cell lines with different lineages: fibroblast-derived cells (3T3 cell...

Mott, Natasha N.; Chung, Wilson C. J.; Tsai, Pei-san; Pak, Toni R.

2010-01-01

213

Fibroblast Growth Factor 22 Is Not Essential for Skin Development and Repair but Plays a Role in Tumorigenesis  

OpenAIRE

Fibroblast Growth Factors play critical roles during development, tissue homeostasis and repair by controlling cell proliferation, survival, migration and differentiation. Of the 22 mammalian FGFs, FGF22, a member of the FGF7/10/22 subfamily, has been shown to have a clear role in synaptogenesis, but its roles in other tissues have not been studied. We have investigated the in vivo functions of FGF22 in mice. Fgf22 null animals were viable, fertile and did not display any obvious abnormalitie...

Jarosz, Monika; Robbez-masson, Luisa; Chioni, Athina-myrto; Cross, Barbara; Rosewell, Ian; Grose, Richard

2012-01-01

214

Repair of chromosome damage induced by X-irradiation during G2 phase in a line of normal human fibroblasts and its malignant derivative  

International Nuclear Information System (INIS)

A line of normal human skin fibroblasts (KD) differed from its malignant derivative (HUT-14) in the extent of cytogenetic damage induced by X-irradiation during G2 phase. Malignant cells had significantly more chromatid breaks and gaps after exposure to 25, 50, or 100 rad. The gaps may represent single-strand breaks. Results from alkaline elution of cellular DNA immediately after irradiation showed that the normal and malignant cells in asynchronous population were equally sensitive to DNA single-strand breakage by X-irradiation. Caffeine or beta-cytosine arabinoside (ara-C), inhibitors of DNA repair, when added directly following G2 phase exposure, significantly increased the incidence of radiation-induced chromatid damage in the normal cells. In contrast, similar treatment of the malignant cells had little influence. Ara-C differed from caffeine in its effects; whereas both agents increased the frequency of chromatid breaks and gaps, only ara-C increased the frequency of gaps to the level observed in the irradiated malignant cells. Addition of catalase, a scavenger of the derivative free hydroxyl radical (.OH), to the cultures of malignant cells before, during, and following irradiation significantly reduced the chromatid damage; and catalase prevented formation of chromatid gaps. The DNA damage induced by X-ray during G2 phase in the normal KD cells was apparently repaired by a caffeine- and ara-C-sensitive mechanism(s) that was deficient or absent in their malignat was deficient or absent in their malignant derivatives

215

Differential regulation of papilloma virus early gene expression in transformed fibroblasts and carcinoma cell lines.  

OpenAIRE

Treatment of bovine papilloma virus (BPV) 1-transformed mouse fibroblasts with cycloheximide led to a 10-fold increase in the amount of viral transcripts, after as little as 1 h of protein synthesis inhibition. Northern blots revealed no qualitative changes in the RNA pattern. Nuclear run-on experiments showed about a 7-fold increase in specific transcriptional activity after cycloheximide treatment. The half-life of BPV1 mRNA was twice as long as in untreated controls. These results indicate...

Kleiner, E.; Dietrich, W.; Pfister, H.

1986-01-01

216

Synthesis and protective effects of bis{4-[N,N-di-(carboxymethyl)amino]phenoxy}alkane derivatives on UVA-induced production of MMP-1 in human skin fibroblasts.  

Science.gov (United States)

UV-induced matrix metalloproteinase (MMP) production is considered a cause of skin aging. In this study, a number of novel bis{4-[N,N-di-(carboxymethyl)amino]phenoxy}alkane derivatives were synthesized and evaluated as UVA-protective agents. These compounds significantly protected human dermal fibroblast (HDF) cells from UVA-induced cytotoxicity and inhibited MMP-1 activation and expression with potency comparable to desferoxamine (DFO). Promoter activity assay indicated that they inhibited MMP-1 expression at the transcriptional level. Further studies revealed that the mechanism of these compounds may include blockage of the UVA-induced activation of the p38/mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK) pathways. Together, these results suggest that further development of these compounds may be of interest. PMID:25177015

Hsu, Ling-Yih; Nien, Chih-Ying; Huang, Wei-Ming; Hsu, Shou-Che; Chang, Tsu-Chung

2014-01-01

217

Relationships of cell proliferation and expression of integrin subunits and type I collagen in skin fibroblasts with renal lesions in patients with IDDM.  

Science.gov (United States)

Previous studies have shown that cultured skin fibroblasts (SFs) from insulin-dependent diabetic mellitus (IDDM) patients with diabetic nephropathy (DN) exhibit both increased proliferation and Na+/H+ antiporter activity. The present study correlated the growth rate and mRNA expression of integrin subunits, extracellular matrix molecules, and transforming growth factor-beta in cultured SFs, with the biopsy determined rate of development of DN lesions ranging from slow to rapid in nine IDDM patients. These varying rates of development of DN lesions were expressed by a mesangial expansion score as estimated by the rate of change in mesangial fraction volume per year. Cultured SF proliferation by direct cell counts positively correlated with mesangial expansion score (r = 0.65; P SFs and the rate of progression of DN lesions may be predictive of the risk to develop clinical DN in IDDM, may be in part genetically regulated, and may be of pathogenetic importance. PMID:9469500

Jin, D K; Kim, Y; Mauer, M; Fioretto, P; Vats, A; Fish, A J

1998-02-01

218

Cutis laxa: reduced elastin gene expression in skin fibroblast cultures as determined by hybridizations with a homologous cDNA and an exon 1-specific oligonucleotide  

International Nuclear Information System (INIS)

Fibroblast cultures were established from six patients with cutis laxa, and elastin gene expression was analyzed by RNA hybridizations with a 2.5-kilobase human elastin cDNA or an exon 1-specific 35-base oligomer. Northern analyses using either probe detected mRNA transcripts of ? 3.5 kilobases, and no qualitative difference between the control and cutis laxa mRNAs was detected. However, quantitation of the elastin mRNA abundance by slot blot hybridizations revealed markedly reduced levels in all cutis laxa cell strains. Assuming equal translational activity of the control and cutix laxa mRNAs, the reduced mRNA levels could result in diminished elastin production, providing an explanation for the paucity of elastic fibers in the skin and other tissues in cutis laxa

219

Effects of PUVA on a human skin epithelial cell line  

International Nuclear Information System (INIS)

An established human epithelial cell line was exposed to photoactivated 8-methoxy psoralen (PUVA) during exponential growth. Effects of PUVA treatment on cell growth were measured by cell kinetic methods (counting of cell numbers, flow cytometric measurements (FCM) of DNA and calculations of labelling indices (LI)). Doses of 8-methoxy psoralen and UVA were comparable to those used in patients. The cell number in PUVA treated cultures remained almost constant, and very few mitoses were seen for 144 h. About 9 h after PUVA, both FCM and LI showed an increase in the fraction of cells in S-phase, reaching a maximum of 85-90% after 24 h. DNA synthesis took place at a low rate in these cells. FCM showed an increasing fraction of polyploid cells after PUVA treatment. The possibility that inhibition of cell proliferation is one of the main effects of PUVA, is discussed. (orig.)

220

Receptor-mediated rapid action of 1 alpha,25-dihydroxycholecalciferol: increase of intracellular cGMP in human skin fibroblasts.  

OpenAIRE

The intracellular cGMP concentration in normal human cultured fibroblasts was increased 2- to 3-fold by 1 alpha,25-dihydroxycholecalciferol [1 alpha,25-(OH)2D3] in a dose-dependent manner between 0.01 nM and 1 microM. The response was detectable within 1 min, reached a maximum (225% +/- 8% of baseline) at 6-8 min, and was no longer detectable at 30 min. The half-maximal effect of 1 alpha,25-(OH)2D3 was at 1.8 nM, and 24,25-dihydroxycholecalciferol showed an estimated EC50 100-fold higher. 1 b...

Barsony, J.; Marx, S. J.

1988-01-01

221

Glycolipid biosurfactants, mannosylerythritol lipids, show antioxidant and protective effects against H(2)O(2)-induced oxidative stress in cultured human skin fibroblasts.  

Science.gov (United States)

Mannosylerythritol lipids (MELs) are biosurfactants known for their versatile interfacial and biochemical properties. To broaden their application in cosmetics, we investigated the antioxidant properties of different MEL derivatives (MEL-A, -B, and -C) by using a 1,1-diphenyl-2-picryl hydrazine (DPPH) free-radical- and superoxide anion-scavenging assay. All MEL derivatives tested showed antioxidant activity in vitro, but at lower levels than those of arbutin. Of the MELs, MEL-C, which is produced from soybean oil by Pseudozyma hubeiensis, showed the highest rates of DPPH radical scavenging (50.3% at 10 mg/mL) and superoxide anion scavenging (>50% at 1 mg/mL). The antioxidant property of MEL-C was further examined using cultured human skin fibroblasts (NB1RGB cells) under H(2)O(2) induced oxidative stress. Surprisingly, MEL-C had a higher protective activity against oxidative stress than arbutin did: 10 µg/mL of MEL-C and arbutin had protective activities of 30.3% and 13%, respectively. Expression of an oxidative stress marker, cyclooxygenase-2, in these cells was repressed by treatment with MEL-C as well as by arbutin. MEL-C was thus confirmed to have antioxidant and protective effects in cells, and we suggest that MELs have potential as anti-aging skin care ingredients. PMID:22864517

Takahashi, Makoto; Morita, Tomotake; Fukuoka, Tokuma; Imura, Tomohiro; Kitamoto, Dai

2012-01-01

222

Morphological features of iPS cells generated from Fabry disease skin fibroblasts using Sendai virus vector (SeVdp).  

Science.gov (United States)

We generated iPS cells from human dermal fibroblasts (HDFs) of Fabry disease using a Sendai virus (SeVdp) vector; this method has been established by Nakanishi et al. for pathogenic evaluation. We received SeVdp vector from Nakanishi and loaded it simultaneously with four reprogramming factors (Klf4, Oct4, Sox2, and c-Myc) to HDFs of Fabry disease; subsequently, we observed the presence of human iPS-like cells. The Sendai virus nucleocapsid protein was not detected in the fibroblasts by RT-PCR analysis. Additionally, we confirmed an undifferentiated state, alkaline phosphatase staining, and the presence of SSEA-4, TRA-1-60, and TRA-1-81. Moreover, ultrastructural features of these iPS cells included massive membranous cytoplasmic bodies typical of HDFs of Fabry disease. Thus, we successfully generated human iPS cells from HDFs of Fabry disease that retained the genetic conditions of Fabry disease; also, these abnormal iPS cells could not be easily differentiated into mature cell types such as neuronal cells, cardiomyocytes, etc. because of a massive accumulation of membranous cytoplasmic bodies in lysosomes, possibly the persistent damages of intracellular architecture. PMID:23810832

Kawagoe, Shiho; Higuchi, Takashi; Otaka, Manami; Shimada, Yohta; Kobayashi, Hiroshi; Ida, Hiroyuki; Ohashi, Toya; Okano, Hirotaka J; Nakanishi, Mahito; Eto, Yoshikatsu

2013-08-01

223

Production of a cloned calf from a fetal fibroblast cell line  

Directory of Open Access Journals (Sweden)

Full Text Available The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibroblasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5% were successfully fused and 24 (28.9% developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8% of which were pregnant on day 35 and 3 (16.7% on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg and sexual behavior (libido and semen characteristics.

Mello M.R.B.

2003-01-01

224

HPV-positive HNSCC cell lines but not primary human fibroblasts are radiosensitized by the inhibition of Chk1  

International Nuclear Information System (INIS)

Purpose: Despite the comparably high cure rates observed for HPV-positive HNSCC, there is still a great need for specific tumor radiosensitization due to the often severe side effects resulting from intense radiochemotherapy. We recently demonstrated that HPV-positive HNSCC cell lines are characterized by a defect in DNA double-strand break repair associated with a pronounced G2-arrest. Here we tested whether abrogation of this radiation-induced G2-arrest by the inhibition of Chk1 results in specific radiosensitization of HPV-positive HNSCC cells. Materials and methods: Experiments were performed with five HPV and p16-positive (93-VU-147T, UM-SCC-47, UT-SCC-45, UD-SCC-2, UPCI-SCC-154) and two HPV and p16-negative HNSCC cell lines, as well as two normal human fibroblast strains. Chk1 was inhibited by the selective inhibitor PF-00477736. Cell cycle distribution was determined by flow cytometry, Chk1-activity via Western blot and cell survival by colony formation assay. Results: With the exception of UPCI-SCC-154, the inhibition of Chk1 was found to abolish the pronounced radiation-induced G2-arrest in all HPV-positive cells utilized. All tumor cell lines that demonstrated the abrogation of G2-arrest also demonstrated radiosensitization. Notably, in G1-arrest-proficient normal human fibroblasts no radiosensitization was induced. Conclusion: Abrogation of the G2 checkpoint through the inhibition of Chk1 may be used to selectively increase the cellular radiosensitivity of HPV-positive HNSCC without affecting the surrounding normal tissue

225

Photoprotective potential of emulsions formulated with Buriti oil (Mauritia flexuosa) against UV irradiation on keratinocytes and fibroblasts cell lines.  

Science.gov (United States)

Considering the belief that natural lipids are safer for topical applications and that carotenoids are able to protect cells against photooxidative damage, we have investigated whether topical creams and lotions, produced with Buriti oil and commercial surfactants, can exert photoprotective effect against UVA and UVB irradiation on keratinocytes and fibroblasts. Cell treatment was divided into two steps, prior and after exposition to 30 min of UVA plus UVB radiation or to 60 min of UVA radiation. Emulsions prepared with ethoxylated fatty alcohols as surfactants and containing alpha-tocopherol caused phototoxic damage to the cells, especially when applied prior to UV exposure. Damage reported was due to prooxidant activity and phototoxic effect of the surfactant. Emulsions prepared with Sorbitan Monooleate and PEG-40 castor oil and containing panthenol as active ingredient, were able to reduce the damages caused by radiation when compared to non-treated cells. When the two cell lines used in the study were compared, keratinocytes showed an increase in cell viability higher than fibroblasts. The Buriti oil emulsions could be considered potential vehicles to transport antioxidants precursors and also be used as adjuvant in sun protection, especially in after sun formulations. PMID:19766688

Zanatta, C F; Mitjans, M; Urgatondo, V; Rocha-Filho, P A; Vinardell, M P

2010-01-01

226

Cytotoxicity of endodontic irrigants containing calcium hydroxide and sodium lauryl sulphate on fibroblasts derived from mouse L929 cell line  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese O objetivo desta pesquisa foi avaliar a citotoxicidade de soluções irrigadoras de canais radiculares contendo hidróxido de cálcio e lauril sulfato de sódio em linhagem de fibroblastos L929. Solução aquosa saturada de hidróxido de cálcio, lauril sulfato de sódio e HCT20 (lauril sulfato de sódio e hid [...] róxido de cálcio) foram diluídos em água destilada em concentrações de 50%, 20%, 10% e 5%. O grupo controle foi representado por meio de cultura de células (MEM - minimum essential medium). A citotoxicidade das soluções sobre os fibroblastos foi avaliada em 4 e 24 h de contato, pelo método do cromo radioativo. Os resultados foram analisados estatisticamente pelo teste do qui-quadrado. Em todas as análises, o intervalo de confiança referente às médias entre os grupos foi estabelecido em 95%. As soluções saturadas de hidróxido de cálcio e o HCT20 apresentaram toxicidade nas concentrações de 50%. O lauril sulfato de sódio foi tóxico em todas as concentrações. As soluções de hidróxido de cálcio em concentrações menores que 50% apresentaram tolerância celular, assim como combinadas ao lauril sulfato de sódio. Tal comportamento não foi observado na solução pura de lauril sulfato de sódio em todas as concentrações. Abstract in english The aim of this study was to evaluate the cytotoxicity of root canal irrigating solutions containing calcium hydroxide and sodium lauryl sulphate on fibroblasts derived from L929 cell line. Saturated calcium hydroxide aqueous solution (CH), sodium lauryl sulphate (SLS) and SLS associated with calciu [...] m hydroxide (HCT20) were diluted with sterile distilled water at 50%, 20%, 10% and 5% concentrations. Minimum essential medium (MEM) served as the control group. The cytotoxicity of the solutions was evaluated on L929 mouse fibroblast cell line, at 4 and 24 h of contact time by the 51Cr radiotracer method. Data were compared and statistical inferences were made with the chi-square test. In all analysis, significance level was set at 5%. CH and HCT20 showed toxicity at 50% concentration, while at concentrations lower than 50% these solutions showed cell tolerance. SLS was cytotoxic at all concentrations. In conclusion, the association of calcium hydroxide and SLS (HCT20) combines the beneficial properties of these solutions and was not harmful to the fibroblast cell line, seeming to be a suitable endodontic irrigating solution.

Sérgio Valmor, Barbosa; Cristiane Maria Sodré, Barroso; Patrícia Alvarez, Ruiz.

227

Establishment of a pig fibroblast-derived cell line for locus-directed transgene expression in cell cultures and blastocysts  

DEFF Research Database (Denmark)

We report the establishment of a spontaneously immortalized pig cell line designated Pig Flip-in Visualize (PFV) for locus-directed transgene expression in pig cells and blastocysts. The PFV cell line was isolated from pig ear fibroblasts transfected with a Sleeping Beauty DNA transposon-based docking vector harbouring a selection gene, an eGFP reporter gene, and an Flp recombinase site for locus-directed gene insertion. PFV cells have insertion of a single docking vector with stable eGFP expression and generated phenotypic normal blastocysts with transgene expression after somatic cell nuclear transfer. PFV cells supported Flp mediated cassette exchange for transgene substitution of eGFP with dsRED, and the dsRED transgenic PFV cells generated blastocysts with transgene expression. Hence, the PFV cell line constitutes a valuable pig equivalent to transformed cell lines from other mammalian species suitable for locus-directed transgene expression in cell cultures and, in addition, for transgene analyses in the very early embryonic stages.

Jakobsen, Jannik E; Li, Juan

2011-01-01

228

Antibacterial Effect of Kenger Gum on Mutans Streptococci and Its Cytotoxic Effect on the 3T3 Fibroblast Cell Line.  

Science.gov (United States)

Purpose: To evaluate the antibacterial effect of Kenger gum on mutans streptococci (in vivo) and Streptococcus mutans (in vitro) and its cytotoxic effect on the 3T3 fibroblast cell line. Materials and Methods: In vitro antibacterial activity of Kenger gum extracts against S.mutans was determined by the disk-diffusion method. The broth dilution method was used to determine the minimum inhibitory concentration (MIC). The cytotoxic effect on 3T3 fibroblast cells at different time intervals was determined using cell culture and viability assays. Clinical studies were then performed on 20 healthy adult subjects, where a sugar-free chewing gum was used as a control. To determine the MS counts, oral rinse samples were taken before chewing as well as 30 and 60 min after 15 min of chewing. Repeated-measures ANOVA was used to compare the bacteria level in the oral rinse samples between the two chewing gums. The Least Significant Difference test was used for adjustment for multiple comparisons. Results: The MIC of the acetone extract of Kenger gum was 30 ?g/ml. The acetone extract of Kenger gum possessed moderate antiproliferative properties against the non-tumorigenic cell line 3T3. A statistically significant decrease was observed for both chewing gums at 30 and 60 min. The decrease continued at 60 min after chewing Kenger gum, while the values for control gum tended to approach the baseline after 60 min. Conclusion: This preliminary study showed that Kenger gum had particular and prolonged antibacterial activity against S. mutans and salivary mutans streptococci. PMID:25197726

Topcuoglu, Nursen; Lacin, Cagdas Caglar; Erguven, Mine; Bilir, Ayhan; Sutlupinar, Nurhayat; Kulekci, Guven

2014-09-01

229

Skin  

International Nuclear Information System (INIS)

Visibility of changes, accessibility for sampling and measuring and extensive past study, make skin the premier model for the assessment of dose-time fractionation relationships. The primary pathophysiologic basis of radiation-induced reactions in the skin is disruption of the reproductive activity of the basal cells. Radiation reactions in the skin increase in proportion to: (a) increased absorption of energy related to the quality of radiation used; (b) increased area irradiated; (c) increased total dose, and (d) decreased overall period of treatment. Radiation-induced reactions may decrease per unit dose with very low dose rates (i.e. 100 cGy/h) and very high dose rates (i.e. 10 Gy/s). Radiation-induced reactions may decrease per unit dose with prolongation of the individual treatment or overall time of the series of treatments. The proportionality between early and late reactions can be altered by various factors such as dose increment size or the quality of the radiations. The most dangerous changes clinically reduce the intensity of the acute reaction in proportion to the late injury

230

Ispaghula (Plantago ovata) seed husk polysaccharides promote proliferation of human epithelial cells (skin keratinocytes and fibroblasts) via enhanced growth factor receptors and energy production.  

Science.gov (United States)

Endogenous carbohydrates, especially oligo- and polysaccharides, participate in the regulation of a broad range of biological activities, e. g., signal transduction, inflammation, fertilisation, cell-cell-adhesion and act as in vivo markers for the determination of cell types. In the present study, water-soluble (WS) and gel-forming polysaccharides (GF) of ispaghula seed husk (Plantago ovata Forsskal, Plantaginaceae) were characterised as neutral and acidic arabinoxylans and tested under in vitro conditions for regulating activities on cell physiology of human keratinocytes and human primary fibroblasts. Only water-soluble polysaccharides exhibited strong and significant effects on cell physiology of keratinocytes and fibroblasts. Proliferation of cells of the spontaneously immortalised keratinocyte cell line HaCaT was significantly up-regulated in a dose-independent manner. Analysis of activated signal pathways by RNA analysis proved an effect of the acidic arabinoxylan on the expression of keratinocyte growth factor (KGF) in HaCaT cells. Differentiation behaviour of normal human keratinocytes (NHK) determined by involucrin was slightly influenced, due to the enhanced cell proliferation, leading to a cell-cell-mediated indirect induction of early differentiation. WS did not influence late differentiation, as determined by keratin K1 and K10 titres. PMID:15678371

Deters, A M; Schröder, K R; Smiatek, T; Hensel, Andreas

2005-01-01

231

Comparison of thermoradiosensitization in two human melanoma cell lines and one fibroblast cell line by concurrent mild hyperthermia and low-dose-rate irradiation  

International Nuclear Information System (INIS)

Two human melanoma cell lines, one radioresistant (Sk-MEL-3) and one radiosensitive (HT-144), and a normal human fibroblast line (AG1522) were evaluated for thermoradiosensitization of low-dose-rate irradiation by concurrent mild hyperthermia (39-41 degrees C). None of the cell lines expressed chronic thermotolerance during heating at 39-41 degrees C. The SK-MEL-3 cells were the most heat sensitive, while AG1522 and HT-144 cells had the same sensitivity at 39 and 40 degrees C but HT-144 cells were more sensitive at 41 degrees C. All cell lines expressed thermal enhancement of radiosensitivity with heating during irradiation which increased with heating temperature. The SK-MEL-3 cells, which were the most resistant to radiation and demonstrated the greatest repair of sublethal damage (SLD) during low-dose-rate irradiation, had the greatest thermal enhancement of radiosensitivity, while the HT144 cells, which were the most sensitive and expressed little repair of SLD during low-dose-rate irradiation, had the smallest thermal enhancement of radiosensitivity. These data show that concurrent mild hyperthermia during low-dose-rate irradiation may be most efficacious in radiation-resistant tumor cells which express resistance through an enhanced capacity for repair of SLD. 24 refs., 5 figs., 1 tab

232

Production of a cloned calf from a fetal fibroblast cell line  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibro [...] blasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5%) were successfully fused and 24 (28.9%) developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8%) of which were pregnant on day 35 and 3 (16.7%) on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg) and sexual behavior (libido and semen characteristics).

M.R.B., Mello; H.V.A., Caetano; M.G., Marques; M.S., Padilha; J.F., Garcia; M.P., Milazzotto; M.E.O.A., Assumpção; A.S., Lima; A.C., Nicácio; C.M., Mendes; V.P., Oliveira; J.A., Visintin.

1485-14-01

233

Performance of a novel keratinocyte-based reporter cell line to screen skin sensitizers in vitro  

International Nuclear Information System (INIS)

In vitro tests are needed to replace animal tests to screen for the skin sensitization potential of chemicals. Skin sensitizers are electrophilic molecules and the Nrf2-electrophile-sensing pathway comprising the repressor protein Keap1, the transcription factor Nrf2 and the antioxidant response element (ARE) is emerging as a toxicity pathway induced by skin sensitizers. Previously, we screened a large set of chemicals in the reporter cell line AREc32, which contains an eight-fold repeat of the rat GSTA2 ARE-sequence upstream of a luciferase reporter gene in the human breast cancer cell line MCF7. This approach was now further developed to bring it closer to the conditions in the human skin and to propose a fully standardized assay. To this end, a luciferase reporter gene under control of a single copy of the ARE-element of the human AKR1C2 gene was stably inserted into HaCaT keratinocytes. A standard operating procedure was developed whereby chemicals are routinely tested at 12 concentrations in triplicate for significant induction of gene activity. We report results from this novel assay on (i) a list of reference chemicals published by ECVAM, (ii) the ICCVAM list of chemicals for validation of alternative endpoints in the LLNA and (iii) on a more general list of 67 chemicals derived from the ICCVAM database. For comparison, peptide reactivity data are presented for the same chemicals. The results indicate a good predictive value of this approach for hazard identifivalue of this approach for hazard identification. Its technical simplicity, the high-throughput format and the good predictivity may make this assay a candidate for rapid validation to meet the tight deadline to replace animal tests for skin sensitization by 2013 set by the European authorities.

234

Benzo(a)pyrene metabolism, DNA-binding and UV-induced repair of DNA damage in cultured skin fibroblasts from a patient with unilateral multiple basal cell carcinoma  

International Nuclear Information System (INIS)

The metabolism of benzo(a)pyrene (BP), and its subsequent binding to DNA, and the repair of UV-induced DNA damage were studied in fibroblasts cultured from the skin of a 61-year-old male who had multiple basal cell carcinoma (BCC) (>100) on his left upper trunk. Results suggest that BP metabolism and repair of DNA are altered in tumor-bearing site (TSB) cells and that patients with this type of metabolic profile may be at higher risk of the development of cutaneous neoplasms. It is also possible that fibroblasts from tumour bearing skin undergo some as yet unexplained alteration in carcinogen metabolism as a consequence of the induction of neoplasia. (author)

235

Both near ultraviolet radiation and the oxidizing agent hydrogen peroxide induce a 32-kDa stress protein in normal human skin fibroblasts  

International Nuclear Information System (INIS)

We have analyzed the pattern of protein synthesis in solar near ultraviolet (334 nm, 365 nm) and near visible (405 nm) irradiated normal human skin fibroblasts. Two hours after irradiation we find that one major stress protein of approximately 32 kDa is induced in irradiated cells. This protein is not induced by ultraviolet radiation at wavelengths shorter than 334 nm and is not inducible by heat shock treatment of these cells. Although sodium arsenite, diamide, and menadione all induced a 32-kDa protein, they also induced the major heat shock proteins. In contrast, the oxidizing agent, hydrogen peroxide, induced the low molecular weight stress protein without causing induction of the major heat shock proteins. A comparison of the 32-kDa proteins induced by sodium arsenite, H2O2, and solar near ultraviolet radiation using chemical peptide mapping shows that they are closely related. These results imply that the pathways for induction of the heat shock response and the 32-kDa protein are not identical and suggest that, at least in the case of radiation and treatment with H2O2, the 32-kDa protein might be induced in response to cellular oxidative stress. This conclusion is supported by the observation that depletion of endogenous cellular glutathione prior to solar near ultraviolet irradiation lowers the fluence threshold for induction of the 32-kDa stress protein

236

Both near ultraviolet radiation and the oxidizing agent hydrogen peroxide induce a 32-kDa stress protein in normal human skin fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

We have analyzed the pattern of protein synthesis in solar near ultraviolet (334 nm, 365 nm) and near visible (405 nm) irradiated normal human skin fibroblasts. Two hours after irradiation we find that one major stress protein of approximately 32 kDa is induced in irradiated cells. This protein is not induced by ultraviolet radiation at wavelengths shorter than 334 nm and is not inducible by heat shock treatment of these cells. Although sodium arsenite, diamide, and menadione all induced a 32-kDa protein, they also induced the major heat shock proteins. In contrast, the oxidizing agent, hydrogen peroxide, induced the low molecular weight stress protein without causing induction of the major heat shock proteins. A comparison of the 32-kDa proteins induced by sodium arsenite, H/sub 2/O/sub 2/, and solar near ultraviolet radiation using chemical peptide mapping shows that they are closely related. These results imply that the pathways for induction of the heat shock response and the 32-kDa protein are not identical and suggest that, at least in the case of radiation and treatment with H/sub 2/O/sub 2/, the 32-kDa protein might be induced in response to cellular oxidative stress. This conclusion is supported by the observation that depletion of endogenous cellular glutathione prior to solar near ultraviolet irradiation lowers the fluence threshold for induction of the 32-kDa stress protein.

Keyse, S.M.; Tyrrell, R.M.

1987-10-25

237

Molecular analysis of gamma-ray-induced mutations at the hprt locus in primary human skin fibroblasts by multiplex polymerase chain reaction  

International Nuclear Information System (INIS)

A total of 153 hprt mutants (23 spontaneous, 130 ?-ray-induced) of primary human skin fibroblasts were isolated and genetic alterations at the locus were studied by multiplex polymerase chain reaction (PCR). The analyses showed that 51% (66/130) of ?-ray-induced genetic changes were large deletions, whereas the majority of spontaneous mutants (21/23) exhibited point mutations. The spectrum of large genetic alterations appeared to be dependent on dose in ?-ray-induced (1-4 Gy) mutations; mutants with complete loss of the hprt locus comprised 21 (3/14) or 39% (15/38) of clones isolated after irradiation with 1 or 4 Gy, respectively. The frequency of partial deletions was found to be higher in the mutants isolated from clones irradiated with 2 Gy (38%) than from those irradiated with 4 Gy (8%). Mapping of all intragenic depletion breakpoints exhibited a nonrandom distribution of breakpoints toward the 3' end of the hprt gene. 46 refs., 4 figs., 2 tabs

238

Maitotoxin activates a nonselective cation channel and a P2Z/P2X(7)-like cytolytic pore in human skin fibroblasts.  

Science.gov (United States)

Maitotoxin (MTX), a potent cytolytic agent, activates Ca(2+) entry via nonselective cation channels in virtually all types of cells. The identity of the channels involved and the biochemical events leading to cell lysis remain unknown. In the present study, the effect of MTX on plasmalemmal permeability of human skin fibroblasts was examined. MTX produced a time- and concentration-dependent increase in cytosolic free Ca(2+) concentration that depended on extracellular Ca(2+) and was relatively insensitive to blockade by extracellular lanthanides. MTX also produced a time- and concentration-dependent increase in plasmalemma permeability to larger molecules as indicated by 1) uptake of ethidium (314 Da), 2) uptake of YO-PRO-1 (375 Da), 3) release of intracellular fura 2 (636 Da), 4) uptake of POPO-3 (715 Da), and, ultimately, 5) release of lactate dehydrogenase (relative molecular weight of 140,000). At the single cell level, uptake of YO-PRO-1 correlated in time with the appearance of large MTX-induced membrane currents carried by the organic cation, N-methyl-D-glucamine (167 Da). Thus MTX initially activates Ca(2+)-permeable cation channels and later induces the formation of large pores. These effects of MTX on plasmalemmal permeability are similar to those seen on activation of P2Z/P2X(7) receptors in a variety of cell types, raising the intriguing possibility that MTX and P2Z/P2X(7) receptor stimulation activate a common cytolytic pore. PMID:10516106

Schilling, W P; Sinkins, W G; Estacion, M

1999-10-01

239

Low-dose radiation activates Nrf1/2 through reactive species and the Ca2+/ERK1/2 signaling pathway in human skin fibroblast cells  

Directory of Open Access Journals (Sweden)

Full Text Available In the current study, we explored the effect of LDR on theactivation of Nrfs transcription factor involved in cellular redoxevents. Experiments were carried out utilizing 0.05 and 0.5 GyX-ray irradiated normal human skin fibroblast HS27 cells. Theresults showed LDR induced Nrf1 and Nrf2 activation andexpression of antioxidant genes HO-1, Mn-SOD, and NQO1.In particular, 0.05 Gy-irradiation increased only Nrf1 activation,but 0.5 Gy induced both Nrf1 and Nrf2 activation.LDR-mediated Nrf1/2 activation was accompanied by reactivespecies (RS generation and Ca2+ flux. This effect was abolishedin the presence of N-acetyl-cysteine and BAPTA- AM.Furthermore, Nrf1/2 activation by LDR was suppressed byPD98059, an inhibitor of ERK1/2. In conclusion, LDR inducesNrf1 and Nrf2 activation and expression of Nrf-regulatedantioxidant defense genes through RS and Ca2+/ERK1/2pathways, suggesting new insights into the molecularmechanism underlying the beneficial role of LDR in HS27cells. [BMB Reports 2013; 46(5: 258-263

Eun Kyeong Lee

2013-05-01

240

Elevated levels of p53 antigen in cultured skin fibroblasts from patients with hereditary adenocarcinoma of the colon and rectum and its relevance to oncogenic mechanisms.  

Science.gov (United States)

Skin fibroblasts (SFs) from patients with adenomatosis of the colon and rectum (ACR) were shown, in general, to express elevated amounts of the p53 antigen as determined by immunoprecipitation with the monoclonal antibody PAb421. Infection with simian virus 40 (SV40) induced expression of the p53 antigen in SFs from normal individuals and further amplified its expression in ACR cells. The half-lives of the p53 antigen in mock-infected ACR cells and in SV40-transformed normal or ACR cells were about 2 and 15 hours, respectively. No immediate differences were detected in the coding segment of the human p53 gene between normal and ACR cells by either Southern or Northern blot analysis. These results suggest that over-expression of the p53 antigen is due, in part, to the increased stability of the p53 antigen in ACR cells. Over-expression represents an early event that is possibly associated with initiation and promotion of inherited colon cancer. PMID:3467115

Kopelovich, L; DeLeo, A B

1986-12-01

241

Growth and Replication of Infectious Bursal Disease Virus in the DF-1 Cell Line and Chicken Embryo Fibroblasts  

Science.gov (United States)

Infectious bursal disease virus (IBDV) causes a highly contagious disease in young chicks and leads to significant economic losses in the poultry industry. To determine a suitable cell line for IBDV infection, replication, and growth kinetics of the virus, DF-1 cells and chicken embryo fibroblasts (CEF) were used. The population doubling per day (Pd/D) was found to be higher in DF-1 as compared to CEF cells. A suitable time of infection (TOI) was established for increased production of virus and greater infectivity titers. The DF-1 and CEF cells were found to be susceptible to infection by producing marked cytopathic effects (CPEs), and the growth curves of IBDV in DF-1 and CEF cells were evaluated by infectivity assay using tissue culture infectious dose (TCID50). The cytopathic effects of the virus in DF-1 and CEF cells were found to be similar, but higher viral titers were detected in the DF-1 cells as compared to CEF. Thus the DF-1 cell line had a higher growth potential and infectivity, which will be of advantage in vaccine production. PMID:24949455

Rekha, Kaliyaperumal; Sivasubramanian, Chandran; Chung, Ill-Min; Thiruvengadam, Muthu

2014-01-01

242

Antagonism of phenanthrene cytotoxicity for human embryo lung fibroblast cell line HFL-I by green tea polyphenols  

Energy Technology Data Exchange (ETDEWEB)

Polycyclic aromatic hydrocarbons (PAHs) have been detected in some commercial teas around the world and pose a threat to tea consumers. However, green tea polyphenols (GTP) possess remarkable antioxidant and anticancer effects. In this study, the potential of GTP to block the toxicity of the model PAH phenanthrene was examined in human embryo lung fibroblast cell line HFL-I. Both GTP and phenanthrene treatment individually caused dose-dependent inhibition of cell growth. A full factorial design experiment demonstrated that the interaction of phenanthrene and GTP significantly reduced growth inhibition. Using the median effect method showed that phenanthrene and GTP were antagonistic when the inhibitory levels were less than about 50%. Apoptosis and cell cycle detection suggested that only phenanthrene affected cell cycle significantly and caused cell death; GTP lowered the mortality of HFL-I cells exposed to phenanthrene; However, GTP did not affect modulation of the cell cycle by phenanthrene. - Green tea polyphenols antagonised cytotoxicity of a low-ring PAH phenanthrene.

Mei Xin [Department of Tea Science, Zhejiang University, Hangzhou 310029 (China); Key Laboratory of Horticultural Plant Growth Development and Biotechnology of Ministry of Agriculture, Zhejiang University, Hangzhou 310029 (China); Wu Yuanyuan; Mao Xiao [Department of Tea Science, Zhejiang University, Hangzhou 310029 (China); Tu Youying, E-mail: youytu@zju.edu.c [Department of Tea Science, Zhejiang University, Hangzhou 310029 (China)

2011-01-15

243

Molybdenum nanoparticles-induced cytotoxicity, oxidative stress, G2/M arrest, and DNA damage in mouse skin fibroblast cells (L929).  

Science.gov (United States)

The present investigation was aimed to study the cytotoxicity, oxidative stress, and genotoxicity induced by molybdenum nanoparticles (Mo-NPs) in mouse skin fibroblast cells (L929). Cells were exposed to different concentrations (1-100?g/ml) of Mo-NPs (size 40nm) for 24 and 48h. After the exposure, different cytotoxicity assays (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide, MTT; neutral red uptake, NRU; and cellular morphology) and oxidative stress markers (lipid peroxidation, LPO; glutathione, GSH; and catalase) were studied. Further, Mo-NPs-induced intracellular reactive oxygen species (ROS) generation, mitochondrial membrane potential (MMP), cell cycle arrest, and DNA damage were also studied. L929 cells treated with Mo-NPs showed a concentration- and time-dependent decrease in cell viability and a loss of the normal cell morphology. The percentage cell viability was recorded as 25%, 42%, and 58% by MTT assay and 24%, 46%, and 56% by NRU assay at 25, 50, and 100?g/ml of Mo-NPs, respectively after 48h exposure. Furthermore, the cells showed a significant induction of oxidative stress. This was confirmed by the increase in LPO and ROS generation, as well as the decrease in the GSH and catalase levels. The decrease in MMP also confirms the impaired mitochondrial membrane. The cell cycle analysis and comet assay data revealed that Mo-NPs induced G2/M arrest and DNA damage in a concentration-dependent manner. Our results demonstrated, for the first time, Mo-NPs induced cytotoxicity, oxidative stress and genotoxicity in L929 cells. Thus, data suggest the potential hazardous nature of Mo-NPs. PMID:25437066

Siddiqui, Maqsood A; Saquib, Quaiser; Ahamed, Maqusood; Farshori, Nida N; Ahmad, Javed; Wahab, Rizwan; Khan, Shams T; Alhadlaq, Hisham A; Musarrat, Javed; Al-Khedhairy, Abdulaziz A; Pant, Aditya B

2015-01-01

244

The common properties and the heterogeneity of dermal fibroblast subpopulations.  

Directory of Open Access Journals (Sweden)

Full Text Available Dermal fibroblasts are a dynamic and diverse population of cells whose functions in skin in many respects remain unknown. Normal adult human skin contains at least three distinct subpopulations of fibroblasts, which occupy unique niches in the dermis. Fibroblasts from each of these niches exhibit distinctive differences when cultured separately. Specific differences in fibroblast histophysiology are evident in papillary dermal fibroblasts, which reside in the superficial dermis, and reticular fibroblasts, which reside in the deep dermis. Both of these subpopulations of fibroblasts differ from the fibroblasts that are associated with hair follicles. Fibroblasts engage in fibroblast-epidermal interactions during hair development and in interfollicular regions of skin. They also play an important role in cutaneous structural transformations.

Makarchuk O.I.

2007-01-01

245

Study on characteristics of in vitro culture and intracellular transduction of exogenous proteins in fibroblast cell line of Liaoning cashmere goat.  

Science.gov (United States)

Establishment of fibroblast cell lines of endangered goat breeds and research on the gene or protein functions based on the cells made a significant contribution to the conservation and utilization of genetic resources. In this study, a fibroblast cell line of Liaoning cashmere goat, frozen in 174 cryovials with 5 × 10(6) cells each, was successfully established from 60 goats ear marginal tissues using explant culture and cryopreservation techniques. Biological analysis of in vitro cultured cell line showed that, the cells were morphologically consistent with fibroblasts; the average viability of the cells was 94.9 % before freezing and 90.1 % after thawing; the growth process of cells was consisted of a lag phase, a logarithmic phase and a plateau phase; cell population doubling time was 65.5 h; more than 90 % of cells were diploid prior to the 6th generation; Neither microbial contamination nor cross-contamination was detected. To determine cell permeability, intracellular path and stability of exogenous proteins during the transduction, a TAT protein transduction domain was fused to the C-terminus of enhanced green fluorescent protein, the established fibroblast cell line was treated with the purified exogenous proteins at various concentrations by adding them to the cell culture media for 1-24 h and assayed cell morphology and protein presence, it was found that the purified exogenous proteins readily entered cells at a concentration of 0.1 mg/ml within 1.5 h and some of them could translocate into nucleus, moreover, the exogenous proteins appeared to be stable inside cells for up to 24 h. PMID:23065271

Hu, P F; Guan, W J; Li, X C; Zhang, W X; Li, C L; Ma, Y H

2013-01-01

246

A Comparison between the Colony Formation of Adult Mouse Spermatogonial Stem Cells in Co cultures with Sertoli and STO (Mouse Embryonic Fibroblast Cell Line)  

OpenAIRE

Objective: The aim of this study was to compare the colony formation of spermatogonialstem cells (SSCs) on sertoli and STO (Mouse embryonic fibroblast cell line) feeder celllayers during a two-week period.Materials and Methods: Initially, sertoli cells and SSCs were isolated from adultmouse testes using a two-step enzymatic digestion and lectin immobilization. Characteristicsof the isolated cells were immunocytochemically confirmed by examiningfor the presence of Oct-4, CDH1, promyelocytic le...

Seyed Morteza Koruji; Mansoureh Movahedin; Syede Momeneh Mohamamadi

2010-01-01

247

Low-concentration methylene blue maintains energy production and strongly improves survival of Leigh syndrome French Canadian skin fibroblasts.  

Science.gov (United States)

Leigh syndrome French Canadian (LSFC) is a recessive disease caused by mutations in the LRPPRC gene (leucine-rich pentatricopeptide repeat containing protein). These mutations induce a cytochrome c oxidase (COX) deficiency resulting in episodes of acute acidotic crisis that will often lead to death. There is no effective treatment. Methylene blue (MB) is a redox dye that increases COX content and activity in vitro and in vivo suggesting that MB could prevent and treat LSFC. In this study, the protective effect of low-concentration MB was tested on two LSFC cell lines, including LSFC-F1, homozygous for the mutation A354V, and LSFC-F2 a compound heterozygous for the mutations A354V and C12775STOP. MB effect on metabolic activity was assessed on both LSFC cells in stable and acidotic conditions. For LSFC-F1, results showed that metabolic activity drastically decline after 96 hours in both conditions but not LSFC-F2 and normal cells. MB completely prevents the decrease of metabolic activity in LSFC-F1. Intracellular ATP content was also measured in both culture media. After 96 hours in acidotic medium, ATP content was almost completely depleted for both LSFC cells. Interestingly, MB completely restores ATP content in LSFC-F1 and LSFC-F2 cells. Finally, MB strongly improves the survival of both LSFC cells. PMID:22202226

Legault, Jean; Larouche, Pierre-Luc; Côté, Isabelle; Bouchard, Line; Pichette, André; Robinson, Brian H; Morin, Charles

2011-01-01

248

Microflora of the penile skin-lined neovagina of transsexual women  

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Full Text Available Abstract Background The microflora of the penile skin-lined neovagina in male-to-female transsexuals is a recently created microbial niche which thus far has been characterized only to a very limited extent. Yet the knowledge of this microflora can be considered as essential to the follow-up of transsexual women. The primary objective of this study was to map the neo-vaginal microflora in a group of 50 transsexual women for whom a neovagina was constructed by means of the inverted penile skin flap technique. Secondary objectives were to describe possible correlations of this microflora with multiple patients' characteristics, such as sexual orientation, the incidence of vaginal irritation and malodorous vaginal discharge. Results Based on Gram stain the majority of smears revealed a mixed microflora that had some similarity with bacterial vaginosis (BV microflora and that contained various amounts of cocci, polymorphous Gram-negative and Gram-positive rods, often with fusiform and comma-shaped rods, and sometimes even with spirochetes. Candida cells were not seen in any of the smears. On average 8.6 species were cultured per woman. The species most often found were: Staphylococcus epidermidis, Streptococcus anginosus group spp., Enterococcus faecalis, Corynebacterium sp., Mobiluncus curtisii and Bacteroides ureolyticus. Lactobacilli were found in only one of 30 women There was no correlation between dilatation habits, having coitus, rinsing habits and malodorous vaginal discharge on the one hand and the presence of a particular species on the other. There was however a highly significant correlation between the presence of E. faecalis on the one hand and sexual orientation and coitus on the other (p = 0.003 and p = 0.027 respectively. Respectively 82%, 58% and 30% of the samples showed an amplicon after amplification with M. curtisii, Atopobium vaginae and Gardnerella vaginalis primer sets. Conclusion Our study is the first to describe the microflora of the penile skin-lined neovagina of transsexual women. It reveals a mixed microflora of aerobe and anaerobe species usually found either on the skin, in the intestinal microflora or in a BV microflora.

Claeys Geert

2009-05-01

249

Establishment and characterization of equine fibroblast cell lines transformed in vivo and in vitro by BPV-1: Model systems for equine sarcoids  

International Nuclear Information System (INIS)

It is now widely recognized that BPV-1 and less commonly BPV-2 are the causative agents of equine sarcoids. Here we present the generation of equine cell lines harboring BPV-1 genomes and expressing viral genes. These lines have been either explanted from sarcoid biopsies or generated in vitro by transfection of primary fibroblasts with BPV-1 DNA. Previously detected BPV-1 genome variations in equine sarcoids are also found in sarcoid cell lines, and only variant BPV-1 genomes can transform equine cells. These equine cell lines are morphologically transformed, proliferate faster than parental cells, have an extended life span and can grow independently of substrate. These characteristics are more marked the higher the level of viral E5, E6 and E7 gene expression. These findings confirm that the virus has an active role in the induction of sarcoids and the lines will be invaluable for further studies on the role of BPV-1 in sarcoid pathology

250

Proteolytic and non-proteolytic activation of keratinocyte-derived latent TGF-?1 induces fibroblast differentiation in a wound-healing model using rat skin.  

Science.gov (United States)

Transforming growth factor-?1 (TGF-?1) reportedly causes the differentiation of fibroblasts to myofibroblasts during wound healing. We investigated the mechanism underlying the activation of latent TGF-?1 released by keratinocytes in efforts to identify promising pharmacological approaches for the prevention of hypertrophic scar formation. A three-dimensional collagen gel matrix culture was prepared using rat keratinocytes and dermal fibroblasts. Stratified keratinocytes promoted the TGF receptor-dependent increase in ?-smooth muscle actin (?-SMA) immunostaining and mRNA levels in fibroblasts. Latent TGF-?1 was found to be localized suprabasally and secreted. ?-SMA expression was inhibited by an anti-?v-integrin antibody and a matrix metalloproteinase (MMP) inhibitor, GM6001. In a two-dimensional fibroblast culture, ?-SMA expression depended on the production of endogenous TGF-?1 and required ?v-integrin or MMP for the response to recombinant latent TGF-?1. In keratinocyte-conditioned medium, MMP-dependent latent TGF-?1 secretion was detected. Applying this medium to the fibroblast culture enhanced ?-SMA production. This effect was decreased by GM6001, the anti-?v-integrin antibody, or the preabsorption of latent TGF-?1. These results indicate that keratinocytes secrete latent TGF-?1, which is liberated to fibroblasts over distance and is activated to produce ?-SMA with the aid of a positive-feedback loop. MMP inhibition was effective for targeting both keratinocytes and fibroblasts in this model. PMID:24492413

Hata, Shozaburo; Okamura, Kazuhiko; Hatta, Mitsutoki; Ishikawa, Hiroyuki; Yamazaki, Jun

2014-01-01

251

Interaction between Leptospiral Lipopolysaccharide and Toll-like Receptor 2 in Pig Fibroblast Cell Line, and Inhibitory Effect of Antibody against Leptospiral Lipopolysaccharide on Interaction  

Science.gov (United States)

Leptospiral lipopolysaccharide (L-LPS) has shown potency in activating toll-like receptor 2 (TLR2) in pig fibroblasts (PEFs_NCC1), and causes the expression of proinflammatory cytokines. However, the stimulation by L-LPS was weak eliciting the function of TLR2 sufficiently in pig innate immunity responses during Leptospira infection. In this study, the immune response of pig embryonic fibroblast cell line (PEFs_SV40) was investigated and was found to be the high immune response, thus TLR2 is the predominate receptor of L-LPS in pig cells. Further, we found a strategy using the antibody against L-LPS, to prevent L-LPS interaction with TLR2 in pig cells which could impact on immune activation. PMID:25557825

Guo, Yijie; Fukuda, Tomokazu; Nakamura, Shuichi; Bai, Lanlan; Xu, Jun; Kuroda, Kengo; Tomioka, Rintaro; Yoneyama, Hiroshi; Isogai, Emiko

2015-01-01

252

Stimulation of MMP-11 (stromelysin-3 expression in mouse fibroblasts by cytokines, collagen and co-culture with human breast cancer cell lines  

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Full Text Available Abstract Background Matrix metalloproteinases (MMPs are central to degradation of the extracellular matrix and basement membrane during both normal and carcinogenic tissue remodeling. MT1-MMP (MMP-14 and stromelysin-3 (MMP-11 are two members of the MMP family of proteolytic enzymes that have been specifically implicated in breast cancer progression. Expressed in stromal fibroblasts adjacent to epithelial tumour cells, the mechanism of MT1-MMP and MMP-11 induction remains unknown. Methods To investigate possible mechanisms of induction, we examined the effects of a number of plausible regulatory agents and treatments that may physiologically influence MMP expression during tumour progression. Thus NIH3T3 and primary mouse embryonic fibroblasts (MEFs were: a treated with the cytokines IL-1?, IL-2, IL-6, IL-8 and TGF-? for 3, 6, 12, 24, and 48 hours; b grown on collagens I, IV and V; c treated with fibronectin, con-A and matrigel; and d co-cultured with a range of HBC (human breast cancer cell lines of varied invasive and metastatic potential. Results Competitive quantitative RT-PCR indicated that MMP-11 expression was stimulated to a level greater than 100%, by 48 hour treatments of IL-1?, IL-2, TGF-?, fibronectin and collagen V. No other substantial changes in expression of MMP-11 or MT1-MMP in either tested fibroblast culture, under any treatment conditions, were observed. Conclusion We have demonstrated significant MMP-11 stimulation in mouse fibroblasts using cytokines, matrix constituents and HBC cell lines, and also some inhibition of MT1-MMP. Our data suggest that the regulation of these genes in the complex stromal-epithelial interactions that occur in human breast carcinoma, is influenced by several mechanisms.

Matthaei Klaus I

2004-07-01

253

Performance of full-pupil line-scanning reflectance confocal microscopy in human skin and oral mucosa in vivo.  

Science.gov (United States)

Point-scanning reflectance confocal microscopes continue to be successfully translated for detection of skin cancer. Line-scanning, with the use of a single scanner and a linear-array detector, offers a potentially smaller, simpler and lower cost alternative approach, to accelerate widespread dissemination into the clinic. However, translation will require an understanding of imaging performance deep within scattering and aberrating human tissues. We report the results of an investigation of the performance of a full-pupil line-scanning reflectance confocal microscope in human skin and oral mucosa, in terms of resolution, optical sectioning, contrast, signal-to-noise ratio, imaging and the effect of speckle noise. PMID:21750780

Larson, Bjorg; Abeytunge, Sanjeewa; Rajadhyaksha, Milind

2011-07-01

254

Down syndrome fibroblasts are hyperresponsive to beta-adrenergic stimulation.  

OpenAIRE

The hormonal response to human skin fibroblasts after exposure to beta-adrenergic agonists, prostaglandin E1 (PGE1), and cholera toxin was monitored by intracellular cyclic AMP accumulation. Down syndrome (DS; trisomy 21) cells had an approximately 10-fold greater response to beta-adrenergic agonists than did either normal diploid skin fibroblasts or other aneusomic fibroblast strains (trisomy 13, 18, and 22). The altered response in DS fibroblasts was specific for beta-adrenergic agonists, b...

Mcswigan, J. D.; Hanson, D. R.; Lubiniecki, A.; Heston, L. L.; Sheppard, J. R.

1981-01-01

255

Degradation of blood-group A glycolipids in cultures of human skin fibroblasts; an approach to investigation of some lysosomal enzyme defects  

International Nuclear Information System (INIS)

Degradation of blood group A glycosphingolipid A-6-2 has been studied by loading experiments in cultured human skin from controls and from patients with inherited lysosomal enzymopathies (?-N-acetylgactosaminidase, ?-fucosidase, ?-glucocerebrosidase, GM1 ?-galactosidase, ceramidase, sap B and sap-precursor deficiencies). In the cell lines from the most unrelated enzymopathies, accumulation of the glycosphingolipids was found corresponding to the inherent enzyme defect. The technique of feeding of radiolabelled glycolipids to cells in culture and analysis of their metabolism was used to examine whether cells from patients with deficiency of ?-N-acetylgactosaminidase (?-NAGA deficiency, Schindler disease). Further we have extended our study with the same probe to other lysosomal enzymophatic cell lines. For this purpose we isolated glycosphingolipid A-6-2 (IV2-?-fucosyl-IV3-?-N-acetylgactosaminylnelactotetraosylceramide) from erythrocyte membrane and labelled it on the ceramide moiety with tritium. The results of this study clearly show that loading experiments in cell culture are a valuable tool to analyze general degradation pathways, assess the physiological significance and the substrate specificity in vivo of the enzymes involved and estimate their residual activities or that of alternative degradation pathways. (authors)

256

Neural cell adhesion molecule mediates contact-dependent inhibition of growth of near-diploid mouse fibroblast cell line m5S/1M  

OpenAIRE

A near-diploid mouse fibroblast cell line m5S/1M used in this study shows a high sensitivity to contact-dependent inhibition of growth, and the addition of EGF causes both morphological change and loss of contact-dependent inhibition of growth. The m5S/1M cell and its transformants obtained by x-ray irradiation have been used to search for the cell surface glycoproteins that are responsible for the growth regulation via cell-cell interactions. Lectin blotting analyses showed that the expressi...

1991-01-01

257

Development of a swine model of secondary liver tumor from a genetically induced swine fibroblast cell line  

OpenAIRE

Aim. Metastatic disease is the most common liver tumor. Although alternative therapies have been developed for non-surgical candidates, those therapies lacked ideal testing prior to clinical application because of a paucity of large animal models. The purpose of the present study was to develop a model for secondary liver tumor in a large animal. Material and methods. Fibroblasts were isolated from swine ear lobules and then transfected with amphotrophic retroviruses encoding human or murine ...

Abbas, R.; Adam, S. J.; Okadal, S.; Groar, H.; Anderson, J.; Sanabria, J.

2008-01-01

258

SOME CORRELATION BETWEEN ONSET OF SPECIFIC DISEASES AND INDICATION SYSTEM IN SKIN AND LINES OF HUMAN PALM  

OpenAIRE

There are various lacuna and voids in clinical studies of glandular disorders and its warning or indication system in the study of palms of human beings. This paper presents for the first time, a datum and several observations made on the conditions of skin, colour mounts, lines etc. of human palms. Studies reveal that palms of both hands play a diagnostic role in medical emergency. The colour, temperature of the skin of hands at times yield more information of impending shock than either pul...

Karnick, C. R.

1987-01-01

259

Effect of CTGF/CCN2 on osteo/cementoblastic and fibroblastic differentiation of a human periodontal ligament stem/progenitor cell line.  

Science.gov (United States)

Appropriate mechanical loading during occlusion and mastication play an important role in maintaining the homeostasis of periodontal ligament (PDL) tissue. Connective tissue growth factor (CTGF/CCN2), a matricellular protein, is known to upregulate extracellular matrix production, including collagen in PDL tissue. However, the underlying mechanisms of CTGF/CCN2 in regulation of PDL tissue integrity remain unclear. In this study, we investigated the effect of CTGF/CCN2 on osteo/cementoblastic and fibroblastic differentiation of human PDL stem cells using the cell line 1-11. CTGF/CCN2 expression in rat PDL tissue and human PDL cells (HPDLCs) was confirmed immunohisto/cytochemically. Mechanical loading was found to increase gene expression and secretion of CTGF/CCN2 in HPDLCs. CTGF/CCN2 upregulated the proliferation and migration of 1-11 cells. Furthermore, increased bone/cementum-related gene expression in this cell line led to mineralization. In addition, combined treatment of 1-11 cells with CTGF/CCN2 and transforming growth factor-?1 (TGF-?1) significantly promoted type I collagen and fibronectin expression compared with that of TGF-?1 treatment alone. Thus, these data suggest the underlying biphasic effects of CTGF/CCN2 in 1-11 cells, inducible osteo/cementoblastic, and fibroblastic differentiation dependent on the environmental condition. CTGF/CCN2 may contribute to preservation of the structural integrity of PDL tissue, implying its potential use as a therapeutic agent for PDL regeneration. PMID:24905848

Yuda, Asuka; Maeda, Hidefumi; Fujii, Shinsuke; Monnouchi, Satoshi; Yamamoto, Naohide; Wada, Naohisa; Koori, Katsuaki; Tomokiyo, Atsushi; Hamano, Sayuri; Hasegawa, Daigaku; Akamine, Akifumi

2015-01-01

260

Photosensitization of human skin cell lines by ATMPn (9-acetoxy-2,7,12,17-tetrakis-(beta-methoxyethyl)-porphycene) in vitro: mechanism of action.  

Science.gov (United States)

9-Acetoxy-2,7,12,17-tetrakis-(beta-methoxyethyl)-porphycene (ATMPn) is a promising new photosensitizer characterized by high absorption around 640 nm and high singlet oxygen yield. To study the mechanism of action in vitro we have investigated uptake, intracellular localization, cell survival and ultrastructural changes following photodynamic treatment in human cell lines derived from the skin (SCL1 and SCL2, squamous cell carcinoma; HaCaT keratinocytes; N1 fibroblasts). Using flow cytometry we have determined the cellular fluorescence as a marker for the uptake of ATMPn after incubation for 60 min. Co-staining with ATMPn and fluorescent dyes specific for cell organelles reveals an intracellular localization of ATMPn in lysosomes. Following irradiation using an incoherent light source (580-740 nm) and a light fluence of 24 J cm-2, phototoxicity is determined by means of the 3-4.5 dimethylthiazol-2,5 diphenyl tetrazolium bromide (MTT) assay. For all cell lines ATMPn concentrations above 15 nM yield a significant phototoxic effect. The 50% effective concentration, EC50, for SCL1 cells is 11.2 +/- 2.9 nM ATMPn. ATMPn uptake and phototoxicity are more effective for HaCaT and SCL1 as compared to SCL2 and N1 cells. Growth curves confirmed the results of the MTT assay. Because of the high lysosomal accumulation of ATMPn, already low photosensitizer concentrations without dark toxicity yield a high photodynamic effect. Immunofluorescence and electron microscopy reveal damage to tonofilaments, plasma membrane and mitochondria, indicating a mechanism unrelated to apoptosis. A dose yielding complete cell killing, as needed for oncological indications, might lead to necrosis, whereas lower sub-lethal doses result in induction of apoptosis. PMID:10205875

Fickweiler, S; Abels, C; Karrer, S; Bäumler, W; Landthaler, M; Hofstädter, F; Szeimies, R M

1999-01-01

261

First cloned swamp buffalo produced from adult ear fibroblast cell.  

Science.gov (United States)

The world's first cloned swamp buffalo (Bubalus bubalis) derived from adult ear skin fibroblast has been reported. Donor fibroblast cells were produced from biopsies taken from adult male ear skin and in vitro matured oocytes obtained from a slaughterhouse were used as cytoplasts. A total of 39 blastocysts and 19 morulae fresh embryos were transferred into 12 recipient buffaloes. Progesterone assays indicated establishment of pregnancy in 10 of the 12 buffaloes (83.3%) after 45 days, with six animals still pregnant at 3 months. One recipient maintained pregnancy to term and naturally delivered a 40 kg male calf after 326 days of gestation. DNA analysis showed that the cloned calf was genetically identical to the donor cells. Genotype analyses, using 12 buffalo microsatellite markers, confirmed that the cloned calf was derived from the donor cell lines. In conclusion, the present study reports, for the first time, the establishment of pregnancy and birth of the first cloned Thai swamp buffalo derived from adult ear skin fibroblast cells. PMID:24804579

Tasripoo, K; Suthikrai, W; Sophon, S; Jintana, R; Nualchuen, W; Usawang, S; Bintvihok, A; Techakumphu, M; Srisakwattana, K

2014-05-01

262

Receptors for NPY and PACAP differ in expression and activity during adipogenesis in the murine 3T3-L1 fibroblast cell line  

Science.gov (United States)

Background and purpose: Neuropeptides are involved in the regulation of food intake in the central nervous system, but they might also act on peripheral fat tissue via neuropeptide receptors. Experimental approach: We investigated the receptor expression and activity of pituitary adenylate cyclase-activating polypeptide (PACAP) and of neuropeptide Y at the mRNA and protein levels in the 3T3-L1 fibroblast line during differentiation into adipocytes. Intracellular calcium concentration was measured by calcium imaging. Key results: The PACAP receptors PAC1 and VPAC2 as well as the neuropeptide Y1 receptor were expressed at the mRNA level in fibroblasts, pre-adipocytes and adipocytes. The mRNA profile of the PAC1 receptor isoforms showed the HOP sequence, whereas the HIP-isoform was present in subconfluent 3T3-L1 fibroblasts only. At the protein level, the mature 3T3-L1 adipocytes produced the PAC1 and Y1 receptors; only the PAC1 receptor showed carbohydrate residues. Both neuropeptides induced an increase of intracellular calcium in mature adipocytes, which was absent in the precursor cells. These changes in calcium were mediated by Y1 and PAC1 receptors as demonstrated by the effects of specific receptor agonists and antagonists. Conclusions and implications: As the PAC1-HOP receptor variant seems to be responsible for PACAP-mediated calcium influx in many cell types, the HOP sequence might also mediate the increase in intracellular calcium in adipocytes. Because a high intracellular calcium level is associated with lipogenesis, peptidergic innervation of adipose tissue might be involved in stress-induced obesity. British Journal of Pharmacology (2009) 157, 620–632; doi:10.1111/j.1476-5381.2009.00164.x; published online 27 April 2009 PMID:19422400

Gericke, Martin T; Kosacka, Joanna; Koch, Daniela; Nowicki, Marcin; Schröder, Thomas; Ricken, Albert M; Nieber, Karen; Spanel-Borowski, Katharina

2009-01-01

263

Inhibition of Mitochondrial Cytochrome c Release and Suppression of Caspases by Gamma-Tocotrienol Prevent Apoptosis and Delay Aging in Stress-Induced Premature Senescence of Skin Fibroblasts  

OpenAIRE

In this study, we determined the molecular mechanism of ?-tocotrienol (GTT) in preventing cellular aging by focusing on its anti-apoptotic effect in stress-induced premature senescence (SIPS) model of human diploid fibroblasts (HDFs). Results obtained showed that SIPS exhibited senescent-phenotypic characteristic, increased expression of senescence-associated ?-galactosidase (SA ?-gal) and promoted G0/G1 cell cycle arrest accompanied by shortening of telomere length with decreased telomera...

Makpol, Suzana; Abdul Rahim, Norhazira; Kien Hui, Chua; Wan Ngah, Wan Zurinah

2012-01-01

264

Modulation of radio-induced oxidative damage by the combination of pentoxifylline and ?-tocopherol in skin fibroblasts and microvascular endothelial cells  

International Nuclear Information System (INIS)

Clinical or accidental localized ionizing radiation exposure can induce severe skin damage constituting the cutaneous radiological syndrome which is divided in acute and late phases. The combination of pentoxifylline (PTX), antioxidant phytochemical, and ?-tocopherol, antioxidant nutrient shows effectiveness in reducing the late radio-induced skin damage with a long period. This work aims to investigate the molecular and cellular mechanisms involved in the effects of this combination

265

Performance of full-pupil line-scanning reflectance confocal microscopy in human skin and oral mucosa in vivo  

OpenAIRE

Point-scanning reflectance confocal microscopes continue to be successfully translated for detection of skin cancer. Line-scanning, with the use of a single scanner and a linear-array detector, offers a potentially smaller, simpler and lower cost alternative approach, to accelerate widespread dissemination into the clinic. However, translation will require an understanding of imaging performance deep within scattering and aberrating human tissues. We report the results of an investigation of ...

Larson, Bjorg; Abeytunge, Sanjeewa; Rajadhyaksha, Milind

2011-01-01

266

Construção de substituto da pele composto por matriz de colágeno porcino povoada por fibroblastos dérmicos e queratinócitos humanos: avaliação histológica / Construction of a skin substitute composed of porcine collagen matrix populated with human dermal fibroblasts and keratinocytes: histological evaluation  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese INTRODUÇÃO: O uso de enxertos autólogos é limitado pela extensão da área doadora e pelo estado clínico dos pacientes, no caso de lesões extensas. Alotransplantes coletados de cadáveres ou voluntários são rejeitados após uma ou duas semanas, servindo apenas como cobertura temporária para essas lesões [...] . O tratamento de grandes lesões cutâneas com tegumento autólogo reconstruído constitui alternativa atraente, já que, a partir de um pequeno fragmento de pele do paciente, pode-se obter culturas de células que se multiplicam rapidamente e podem ser criopreservadas, permitindo, assim, sua utilização em novos tratamentos por tempo indeterminado. Este estudo pretendeu avaliar o comportamento histológico de queratinócitos e fibroblastos humanos cultivados sobre uma matriz de colágeno porcino derivada da submucosa intestinal. MÉTODO: Células da epiderme e derme humana foram cultivadas separadamente e semeadas sobre matriz de colágeno porcino, onde permaneceram em ambiente controlado por 21 dias, antes de serem submetidas a análise histológica. RESULTADOS: Observou-se que os fibroblastos invadem e colonizam a matriz de colágeno, enquanto os queratinócitos se organizam de forma laminar e estratificada sobre a superfície em que foram semeados. CONCLUSÕES: A utilização da matriz de colágeno porcino como carreador de células da pele humana é possível e a organização dessas células se assemelha à arquitetura da pele humana. Abstract in english BACKGROUND: In the case of extensive lesions, the use of autologous grafts is limited by the extent of the donor area and the clinical condition of patients. Allografts collected from cadavers or volunteers are usually rejected after 1 to 2 weeks, thus serving only as temporary cover for these lesio [...] ns. Treating major cutaneous lesions with reconstructed autologous skin is an attractive alternative, because it is possible to obtain cultures of cells that multiply rapidly and can be cryopreserved from a small fragment of the patient's skin, thereby facilitating its indefinite use in new treatments. This study evaluated the histological behavior of cultured human keratinocytes and fibroblasts on a collagen matrix derived from porcine small intestinal submucosa. METHODS: Cells from human epidermis and dermis were grown separately and seeded on porcine collagen matrix, which was maintained in a controlled environment for 21 days before being subjected to histological analysis. RESULTS: Fibroblasts invaded and colonized the collagen matrix, whereas keratinocytes were organized in laminated and stratified layers on the surface on which they were seeded. CONCLUSIONS: The use of porcine collagen matrix as a support for human skin cells is feasible, and the organization of these cells resembles the architecture of human skin.

Cesar, Isaac; Francinni M. P., Rego; Pedro Ribeiro Soares de, Ladeir; Silvana C., Altram; Renata C. de, Oliveira; Johnny L. C. B., Aldunate; André O., Paggiaro; Marcus Castro, Ferreira.

2012-12-01

267

PAMPs and DAMPs stimulate the expression of pro-inflammatory cytokines in vitro in a fibroblast cell-line from rainbow trout (Oncorhynchus mykiss)  

DEFF Research Database (Denmark)

The recognition of PAMPs by immune cells relies on conserved PRRs such as TLRs, NLRs and RLRs leading to activation of NF?B signalling pathways. These receptors are activated upon stimulation by different ligands such as bacterial or viral components. The binding of ligands to the receptors activates downstream signalling pathways, which subsequently leads to expression of pro-inflammatory cytokines and chemokines. DAMPs released from necrotic cells may also bind to and activate similar downstream signalling events. In telosts was found that mechanical damage of the muscle tissue using sterile needles induced a very rapid expression of the pro-inflammatory cytokines IL-1?, IL-8 and IL-10 as measured by real-time PCR. The results imply that cells located in the muscular tissue in addition to recruited cells are involved in the observed increased cytokine / chemokine expression. It is believed that this expression to a large extend is mediated by fibroblasts in the musculature. To investigate this, a fibroblastcell-line (RTHDF1) from the rainbow trout was stimulated with either LPS from E. coli, cell debris or supernatant from sonicated fibroblasts. Whereas LPS stimulation resulted in a significant up-regulation of the expression of IL-1?, IL-8 and IL-10 and stimulation with supernatant from sonicated cells led to a significant up-regulation of IL-1? and IL-10, while debris only stimulated the expression of IL-1?. TLR-2 and -4 are not described from salmonid fishes, however TLR-3, -5 and -9 are described in this evolutionary lineage of the bony fishes. The expression of TLR-3 and -9 receptors were significantly up-regulated following physical damage of muscle tissue as well as in stimulated fibroblasts, where LPS induced both TLR-3 and -9, supernatant from sonicated cells only TLR-9 while debris caused no induction. The present study reinforce the idea that fibroblasts are able to react to PAMPs and DAMPs and that non-immune cell-types play an important role in the inflammatory reaction per se. From an evolutionary perspective the facilitation of an inflammatory response through recognition of PAMPs and DAMPs by non immune cells seems plausible.

Ingerslev, Hans-Christian; Ossum, C.G.

268

Case report 511: Fibroblastic rheumatism  

International Nuclear Information System (INIS)

We report a ten-year-old child with the newly described entity of fibroblastic rheumatism. This child developed rapid, progressive, symmetrical polyarthritis, similar to the radiographic appearance of juvenile rheumatoid arthritis, except for the rapidity of progression. The polyarthritis was preceded by the development of skin nodules with characteristic histological changes. (orig./GDG)

269

Organotypic cocultures as skin equivalents: A complex and sophisticated in vitro system  

OpenAIRE

To assess the role of genes required for skin organogenesis, tissue regeneration and homeostasis, we have established in vitro skin equivalents composed of primary cells or cell lines, respectively. In these organotypic cocultures keratinocytes generate a normal epidermis irrespective of the species and tissue origin of fibroblasts. The combination of cells derived from mouse and human tissues facilitates the identification of the origin of compounds involved in epidermal tissue reconstitu...

Stark Hans-Jürgen; Szabowski Axel; Fusenig Norbert E.; Maas-Szabowski Nicole

2004-01-01

270

Direct contact of fibroblasts with neuronal processes promotes differentiation to myofibroblasts and induces contraction of collagen matrix in vitro.  

Science.gov (United States)

Wound healing is often delayed in the patients whose sensory and autonomic innervation is impaired. We hypothesized that existence of neurites in the skin may promote wound healing by inducing differentiation of fibroblasts into myofibroblasts with consequent wound contraction. In the current study, we examined the effect of neurons on differentiation of fibroblasts and contraction of collagen matrix in vitro using a new co-culture model. Neuronal cell line, PC12 cells, of which the neurite outgrowth can be controlled by adding nerve growth factor, was used. Rat dermal fibroblasts were co-cultured with PC12 cells extending neurites or with PC12 cells lacking neurites. Then, differentiation of fibroblasts into myofibroblasts and contraction of the collagen matrix was evaluated. Finally, we examined whether direct or indirect contact with neurites of PC12 cells promoted the differentiation of fibroblasts. Our results showed that fibroblasts co-cultured with PC12 extending neurites differentiated into myofibroblasts more effectively and contracted the collagen matrix stronger than those with PC12 lacking neurites. Direct contact of fibroblasts with neurites promoted more differentiation than indirect contact. In conclusion, direct contact of fibroblasts with neuronal processes is important for differentiation into myofibroblasts and induction of collagen gel contraction, leading to promotion of wound healing. PMID:23758129

Fujiwara, Toshihiro; Kubo, Tateki; Kanazawa, Shigeyuki; Shingaki, Kenta; Taniguchi, Manabu; Matsuzaki, Shinsuke; Gurtner, Geoffrey C; Tohyama, Masaya; Hosokawa, Ko

2013-01-01

271

[Isolation and characteristics of the fibroblast cell line of rats cotransformed by the DNA of simian adenovirus SA7 (C8) and hepatitis B virus].  

Science.gov (United States)

Transformed cell line (SH2) was established by means of co-transfection of primary rat embryonal fibroblasts by DNA of high-oncogenic simian adenovirus SA7 (C-8) and hepatitis B virus. SH2 cells possess a transformed phenotype and high oncogenicity both for allogenic (rats) and xenogenic (hamsters) animals. 100 SH2 cells induce tumours in newborn (3 day) hamsters. 10(4) SH2 cells inoculated to adult hamsters induce tumours in approximately 50% cases on the 20th-30th day. Possible mechanisms of significant stimulation of SH2 cells oncogenic properties after co-transfection by DNA of oncogenic adenovirus SA7 and hepatitis B virus are discussed. PMID:6510333

Potebnia, G P; Naroditski?, B S; Ponomareva, T I; Gartel', A L; Struk, V I

1984-01-01

272

Cytotoxic Evaluation of Elastomeric Dental Impression Materials on a Permanent Mouse Cell Line and on a Primary Human Gingival Fibroblast Culture  

Directory of Open Access Journals (Sweden)

Full Text Available The need for clinically relevant in vitro tests of dental materials is widely recognized. Nearly all dental impression materials are introduced into the mouth just after mixing and allowed to set in contact with the oral tissues. Under these conditions, the materials may be toxic to cells or may sensitize the tissues. The aim of the present study is to evaluate the potential cytotoxicity of new preparations of elastomeric dental impression materials: A four vinylpolysiloxanes: Elite H-D Putty and Elite H-D Light Body (Zhermack, Badia Polesine, Rovigo, Italy; Express Putty and Express Light Body (3M ESPE AG Seefeld, Germany and B two polyethers: Impregum Penta and Permadyne Penta L (3M ESPE AG Seefeld, Germany. The cytotoxicity of these impression materials were examined using two different cell lines: Balb/c 3T3 (permanent cell line and human gingival fibroblasts (primary cell line and their effects were studied by indirect and direct tests. The direct tests are performed by placing one sample of the impression materials in the centre of the Petri dishes at the time of the seeding of cells. The cell growth was evaluated at the 12th and 24th hours by cell number. The indirect tests were performed by incubating a square of 1 cm diameter impression material in 5 mL of medium at 37 °C for 24 hours (“eluates”. Subconfluent cultures are incubated with “eluates” for 24 hours. The MTT-formazan production is the method used for measuring the cell viability. The results indicate that: a polyether materials are cytotoxic under both experimental conditions; b among vinylpolysiloxanes, only Express Light Body (3M ESPE AG Seefeld, Germany induces clear inhibition of cellular viability of Balb/c 3T3 evaluated by direct and indirect tests and c the primary cell line is less sensitive to the toxic effect than the permanent cell line.

Roberta Tiozzo

2009-08-01

273

The potentiation by caffeine of X-ray damage to cultured human skin fibroblasts from normal subjects and ataxia-telangiectasia patients  

International Nuclear Information System (INIS)

Caffeine was found to potentiate X-ray-induced killing of human diploid fibroblasts from a normal subject and an ataxia-telangiectasia (AT) patient when it was present at 2 mM concentration for 30 to 66 hr postirradiation. The dose-modifying factor for caffeine-treated normal cells had an average value of 1.26 +/- 0.13 which did not vary significantly with treatment time or X-ray dose. The dose-modifying factor for caffeine-treated AT cells was 1.12 +/- 0.12 at 30 hr, rose to 1.66 +/- 0.17 at 41 hr, and decreased to 1.31 +/- 0.13 at 66 hr. Thus no clear difference was observed between these two cell strains' susceptibility to postirradiation caffeine treatment

274

ER-? agonist induces conversion of fibroblasts into myofibroblasts, while ER-? agonist increases ECM production and wound tensile strength of healing skin wounds in ovariectomised rats.  

Science.gov (United States)

Oestrogen deprivation is one of the major factors responsible for many age-related processes, including poor wound healing in women. Previously, it has been shown that oestrogens have a modulatory effect in different wound-healing models. Therefore, in this study, the effect of selective oestrogen receptor (ER) agonists (PPT - ER-? agonist, DPN - ER-? agonist) on excisional and incisional wound-healing models was compared in ovariectomised rats in vivo as well as on human dermal fibroblasts (HDF) and human umbilical endothelial cells (HUVEC) in vitro. In the in vivo study, 4 months after either ovariectomy or sham ovariectomy, Sprague-Dawley rats were randomly divided into four groups and subjected to two incisional and excisional wounds: (i) control - sham operated, vehicle-treated; (ii) ovariectomised, vehicle-treated; (iii) ovariectomised, PPT treated; (iv) ovariectomised, DPN treated. In the in vitro study, HDFs and HUVECs were used. After treatment with ER agonists, cells were processed for immunocytochemistry and gelatin zymography. Our study shows that stimulation of ER-? leads to the differentiation of fibroblasts into myofibroblasts both in vivo and in vitro. On the other hand, the formation of extracellular matrix was more prominent, and wound tensile strength (TS) was increased when ER-? was stimulated. In contrast, stimulation of ER-? led to a more prominent increase in the expression of MMP-2 and decrease in wound TS. New information is presented in this investigation concerning oestrogen replacement therapy (ERT) in different wound-healing models. This study demonstrates that the ERT should be both wound and receptor-type specific. PMID:21507066

Novotný, Martin; Vasilenko, Tomáš; Varinská, Lenka; Smetana, Karel; Szabo, Pavol; Sarišský, Marek; Dvo?ánková, Barbora; Mojžiš, Ján; Bobrov, Nikita; Toporcerová, Silvia; Sabol, Franitšek; Matthews, Bryan J O; Gál, Peter

2011-09-01

275

Mutation spectrum of 1,2-dibromo-3-chloropropane, an endocrine disruptor, in the lacI transgenic Big Blue Rat2 fibroblast cell line.  

Science.gov (United States)

1,2-Dibromo-3-chloropropane (DBCP), a soil fumigant against nematodes, has been extensively studied for genotoxicity, carcinogenicity and damage to male reproduction-related organs, as a possible endocrine disruptor. However, the precise mechanisms involved in DBCP-induced mutagenesis and carcinogenesis are as yet unknown. Thus, in this study the mutagenicity and mutation spectrum of DBCP was determined using the lacI transgenic Big Blue Rat2 fibroblast cell line. In determining the optimal concentration of DBCP in Big Blue Rat2 fibroblast cells, the 50% inhibition concentration was calculated to be 0.75 mM. When cells were exposed to DBCP concentrations of 0.21, 0.39 and 0.75 mM, the respective relative survival rates were approximately 80, 70 and 50%. The mean mutant frequencies (MFs) (x 10(-5), +/- SEM) of the medium and 1% DMSO solvent controls were determined as 6.43 +/- 0.616 and 5.28 +/- 1.086, respectively. The MFs (x 10(-5), +/- SEM) of cells exposed to 0.21, 0.39 and 0.75 mM DBCP were 8.09 +/- 1.02, 10.86 +/- 2.17 and 12.26 +/- 0.79, respectively, with a dose-dependent effect (ANOVA, P = 0.007). Moreover, MF values for the 0.75 and 0.39 mM DBCP-treated groups were statistically significant (ANOVA, P DBCP-induced groups. Among 31 single base pair substitutions, 25 (62.5%) occurred at G:C base pairs, while six (15%) were at A:T base pairs. The predominant mutation was G:C-->A:T transitions (16/40, 40%), followed by G:C-->T:A transversions (9/40, 22.5%). We conclude that DBCP is a possible base substitution mutagen, especially at guanine bases. PMID:12110625

Ryu, Jae-Chun; Kim, Youn-Jung; Chai, Young-Gyu

2002-07-01

276

Undifferentiated propagation of the human embryonic stem cell lines, H1 and HSF6, on human placenta-derived feeder cells without basic fibroblast growth factor supplementation.  

Science.gov (United States)

In order for human embryonic stem cells (hESCs) to be cultured on mouse embryonic fibroblast (MEFs) feeder cells, continuous basic fibroblast growth factor (bFGF) supplementation is required. However, the role of bFGF in a culture system using human-derived feeder cells has not been evaluated until now. In this study, we propagated the widely used hESC lines, H1 and HSF6, on human placenta-derived feeder cells (HPCs) without exogenous bFGF supplementation, and were able to propagate hESCs on HPC feeders up to 50 passages. The absence of bFGF in culture media did not interrupt the undifferentiated propagation and the expression of pluripotent stem cell markers ALP, SSEA-4, TRA-60, Oct-4, Nanog, and Rex-1, as well as the formation of embryoid bodies (EBs) and their differentiation potential. In contrast, hESCs cocultured with MEF feeders could not propagate and form EBs without exogenous bFGF supplementation. Expression of bFGF and the activation of the ERK1/2-c-Fos/c-Jun pathway, which is known as the signaling pathway of bFGF, were identifiable not only in hESCs cultured in bFGF-containing media regardless of feeder cell type, but also in hESCs cocultured with HPC feeder cells in media without bFGF. These findings may support the hypothesis that HPC feeder cells enhance endogenous bFGF production and activation of the ERK1/2-c-Fos/c-Jun pathway, which suggests that HPCs have an additional advantage in their hESC propagation compared with MEF. PMID:20201681

Park, Yong; Choi, In Young; Lee, Seung Jin; Lee, Se Ryeon; Sung, Hwa Jung; Kim, Jong Hoon; Yoo, Young Do; Geum, Dong Ho; Kim, Sun Haeng; Kim, Byung Soo

2010-11-01

277

Granulocyte macrophage colony stimulating factor (GM-CSF biological actions on human dermal fibroblasts  

Directory of Open Access Journals (Sweden)

Full Text Available Fibroblasts are involved in all pathologies characterized by increased ExtraCellularMatrix synthesis, from wound healing to fibrosis. Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF is a cytokine isolated as an hemopoietic growth factor but recently indicated as a differentiative agent on endothelial cells. In this work we demonstrated the expression of the receptor for GM-CSF (GMCSFR on human normal skin fibroblasts from healthy subjects (NFPC and on a human normal fibroblast cell line (NHDF and we try to investigate the biological effects of this cytokine. Human normal fibroblasts were cultured with different doses of GM-CSF to study the effects of this factor on GMCSFR expression, on cell proliferation and adhesion structures. In addition we studied the production of some Extra-Cellular Matrix (ECM components such as Fibronectin, Tenascin and Collagen I. The growth rate of fibroblasts from healthy donors (NFPC is not augmented by GM-CSF stimulation in spite of increased expression of the GM-CSFR. On the contrary, the proliferation of normal human dermal fibroblasts (NHDF cell line seems more influenced by high concentration of GM-CSF in the culture medium. The adhesion structures and the ECM components appear variously influenced by GM-CSF treatment as compared to fibroblasts cultured in basal condition, but newly only NHDF cells are really induced to increase their synthesis activity. We suggest that the in vitro treatment with GM-CSF can shift human normal fibroblasts towards a more differentiated state, due or accompanied by an increased expression of GM-CSFR and that such “differentiation” is an important event induced by such cytokine.

S Montagnani

2009-12-01

278

Collagen Fragments Inhibit Hyaluronan Synthesis in Skin Fibroblasts in Response to Ultraviolet B (UVB): NEW INSIGHTS INTO MECHANISMS OF MATRIX REMODELING*  

OpenAIRE

UVB irradiation causes characteristic features of skin aging including remodeling of the dermal extracellular matrix. A key feature during this process is the up-regulation of matrix metalloproteinases and cleavage of collagen. Hyaluronic acid (HA), a major component of the dermal matrix, decreases after chronic UVB exposure. However, the factors that govern the decline of HA synthesis during the course of actinic aging are largely unknown. The aim of the present study was to explore whether ...

Ro?ck, Katharina; Grandoch, Maria; Majora, Marc; Krutmann, Jean; Fischer, Jens W.

2011-01-01

279

A new technique for the transient simulation of transmission lines inclusive of skin effect  

OpenAIRE

A new approach is presented for the transient simulation of lossy transmission lines in high-speed circuits. The approach is based on developing a model for the transmission line which is structured around natural modes of oscillation unlike other transmission-line models which are based on travelling waves. The principal advantage of the new approach is that conversion of frequency-domain prototype models for the transmission lines to the time domain for use in circuit simulators is particul...

Condon, Marissa

2001-01-01

280

Somatic copy-number mosaicism in human skin revealed by induced pluripotent stem cells  

OpenAIRE

Reprogramming human somatic cells into induced pluripotent stem cells (iPSCs) has been suspected of causing de novo copy number variations (CNVs)1-4. To explore this issue, we performed a whole-genome and transcriptome analysis of 20 human iPSC lines derived from primary skin fibroblasts of 7 individuals using next-generation sequencing. We find that, on average, an iPSC line manifests two CNVs not apparent in the fibroblasts from which the iPSC was derived. Using qPCR, PCR, and digital dropl...

Abyzov, Alexej; Mariani, Jessica; Palejev, Dean; Zhang, Ying; Haney, Michael Seamus; Tomasini, Livia; Ferrandino, Anthony; Belmaker, Lior A. Rosenberg; Szekely, Anna; Wilson, Michael; Kocabas, Arif; Calixto, Nathaniel E.; Grigorenko, Elena L.; Huttner, Anita; Chawarska, Katarzyna

2012-01-01

281

The role of opioid receptors in migration and wound recovery in vitro in cultured human keratinocytes and fibroblasts.  

Science.gov (United States)

We have previously described significant changes in skin differentiation and the delay in wound healing from delta-opioid receptor knockout mice. In addition, we have shown that opioid receptor ligands and their receptor systems affect wound healing in vitro and the migration pattern of human skin cells, such as keratinocytes and fibroblasts (Bigliardi-Qi et al., Differentiation 74:174-185, 2006; Bigliardi et al., Exp Dermatol 18:424-430, 2009; Bigliardi et al., J Recept Signal Transduct Res 22:191-199, 2002). This observation is true for both primary keratinocytes and fibroblasts derived from foreskin or normal human skin as well as for immortalized cell lines such as HaCaT cells. PMID:25293334

Bigliardi-Qi, Mei; Bigliardi, Paul L

2015-01-01

282

CopA3 Peptide Prevents Ultraviolet-Induced Inhibition of Type-I Procollagen and Induction of Matrix Metalloproteinase-1 in Human Skin Fibroblasts  

OpenAIRE

Ultraviolet (UV) exposure is well-known to induce premature aging, which is mediated by matrix metalloproteinase-1 (MMP-1) activity. A 9-mer peptide, CopA3 (CopA3) was synthesized from a natural peptide, coprisin, which is isolated from the dung beetle Copris tripartitus. As part of our continuing search for novel bioactive natural products, CopA3 was investigated for its in vitro anti-skin photoaging activity. UV-induced inhibition of type-I procollagen and induction of MMP-1 were partially ...

Dong-Hee Kim; Han-Hyuk Kim; Hyeon-Jeong Kim; Hyun-Gug Jung; Jae-Myo Yu; Eun-Su Lee; Yong-Hun Cho; Dong-In Kim; Bong-Jeun An

2014-01-01

283

Replication and phenotypic expression of control and scleroderma human fibroblasts: responses to growth factors.  

OpenAIRE

To explore the mechanism of increased collagen synthesis by scleroderma skin fibroblasts in vitro, control and scleroderma fibroblasts were compared in confluent monolayer cultures growth-arrested by serum deprivation; responses to optimal mitogenic doses of platelet-derived growth factor, fibroblast growth factor, epidermal growth factor and nerve growth factor were compared. Platelet-derived growth factor had a selective mitogenic effect on control skin fibroblasts not observed with sclerod...

Leroy, E. C.; Mercurio, S.; Sherer, G. K.

1982-01-01

284

Neoplastic transformation of human diploid fibroblasts treated with chemical carcinogens and Co-60 ?-rays  

International Nuclear Information System (INIS)

Two fibroblast cell strains derived from human embryonic lungs (WI-38 and IMR-90) were transformed into neoplastic cells by treatment with Co-60 ?-rays. Four other fibroblast cell strains (two from human embryonic liver and the other two from human adult skin) were transformed into neoplastic cells by treatment with 4-nitroquinoline 1-oxide (4NQO). The transformation was obtained by repeated treatments with these carcinogenic agents, but not by a single treatment in a variety of experimental conditions. These results suggest that transformation of normal human cells might be a multistep process. All of the transformed cell lines had the following characteristics: 1) epithelial-like morphology; 2) unlimited growth potential; 3) abnormal karyotype; 4) increased saturation cell density; 5) low serum requirement for growth; 6) elevated colony formation in soft agar; 7) growth capability in theophylline containing medium; 8) increase of the B(H) subunit of lactate dehydrogenase (LDH) isozyme; and 9) loss of large external transformation sensitive (LETS) protein. The first three characteristics (morphological changes, unlimited growth and abnormal karyotype) are proposed to be sufficient to conclude that neoplastic transformation of normal human fibroblasts has occurred. In order to conduct quantitative transformation experiments with human fibroblasts, criteria of the morphology of transformed colonies were defined. Advantages and disadvantages in the use of normal human disadvantages in the use of normal human fibroblasts for transformation studies are discussed. Finally, future problems in transformation of human cells are described. (J.P.N.)

285

Manganese superoxide dismutase: Effect of the ala16val polymorphism on protein, activity, and mRNA levels in human breast cancer cell lines and stably transfected mouse embryonic fibroblasts  

OpenAIRE

The manganese superoxide dismutase (MnSOD) ala16val polymorphism has been associated with various diseases including breast cancer. In the present study, we investigated levels of MnSOD protein, enzymatic activity and mRNA with respect to MnSOD genotype in several human breast carcinoma cell lines and in mouse embryonic fibroblasts (MEF), developed from the MnSOD knockout mouse, stably expressing human MnSOD-ala and MnSOD-val. In human breast cell lines, the MnSOD-ala allele was associated wi...

Mcatee, Britt L.; Yager, James D.

2010-01-01

286

Cholesteatoma Fibroblasts Promote Epithelial Cell Proliferation through Overexpression of Epiregulin  

OpenAIRE

To investigate whether keratinocytes proliferate in response to epiregulin produced by subepithelial fibroblasts derived from middle ear cholesteatoma. Tissue samples were obtained from patients undergoing tympanoplasty. The quantitative polymerase chain reaction and immunohistochemistry were performed to examine epiregulin expression and localization in cholesteatoma tissues and retroauricular skin tissues. Fibroblasts were cultured from cholesteatoma tissues and from normal retroauricular s...

Yoshikawa, Mamoru; Kojima, Hiromi; Yaguchi, Yuichiro; Okada, Naoko; Saito, Hirohisa; Moriyama, Hiroshi

2013-01-01

287

Skin pigmentary anomalies and mosaicism for an acentric marker chromosome originating from 3q  

OpenAIRE

We report on a 22 year old man with hyperpigmentation distributed along the lines of Blaschko in whom cytogenetic analysis showed mosaicism for an unusual supernumerary marker chromosome. The patient was of normal intelligence and was not dysmorphic. The marker was present in 30% of his lymphocytes and in 6% of his skin fibroblasts from a dark area, while fibroblasts from a light area showed a normal karyotype, 46,XY.We have identified the origin of the marker using fluorescence in situ hybr...

Portnoi, M.; Boutchnei, S.; Bouscarat, F.; Morlier, G.; Nizard, S.; Dersarkissian, H.; Crickx, B.; Nouchy, M.; Taillemite, J.; Belaich, S.

1999-01-01

288

On-line diffusion profile of a lipophilic model dye in different depths of a hair follicle in human scalp skin.  

Science.gov (United States)

In skin and hair research, drug targeting to the hair follicle is of great interest in the treatment of skin diseases. The aim of this study is to visualize on-line the diffusion processes of a model fluorophore into the hair follicle at different depths using fresh human scalp skin and confocal laser scanning microscopy. Up to a depth of 500 microm in the skin, a fast increase of fluorescence is observed in the gap followed by accumulation of the dye in the hair cuticle. Penetration was also observed via the stratum corneum and the epidermis. Little label reached depths greater than 2000 microm. Fat cells accumulated the label fastest, followed by the cuticular area and the outer root sheath of the hair follicle. Sweat glands revealed very low staining, whereas the bulb at a depth of 4000 microm was visualized only by autofluorescence. From this study, we conclude that on-line visualization is a promising technique to access diffusion processes in deep skin layers even on a cellular level. Furthermore, we conclude that the gap and the cuticle play an important role in the initial diffusion period with the label in the cuticle originating from the gap. PMID:16185278

Grams, Ylva Y; Whitehead, Lynne; Lamers, Gerda; Sturmann, Nico; Bouwstra, Joke A

2005-10-01

289

A Comparison between the Colony Formation of Adult Mouse Spermatogonial Stem Cells in Co cultures with Sertoli and STO (Mouse Embryonic Fibroblast Cell Line  

Directory of Open Access Journals (Sweden)

Full Text Available Objective: The aim of this study was to compare the colony formation of spermatogonialstem cells (SSCs on sertoli and STO (Mouse embryonic fibroblast cell line feeder celllayers during a two-week period.Materials and Methods: Initially, sertoli cells and SSCs were isolated from adultmouse testes using a two-step enzymatic digestion and lectin immobilization. Characteristicsof the isolated cells were immunocytochemically confirmed by examiningfor the presence of Oct-4, CDH1, promyelocytic leukaemia zinc finger factor (PLZF,SSC C-kit, and the distribution of Sertoli cell vimentin. SSCs were then cultured abovethe Sertoli, STO and the control (without co-culture separately for two weeks. In allthree groups, the number and diameter of colonies were evaluated using an invert microscopeon the 3rd, 7th, 10th and 14th day. ?1 and ?6-integrin m-RNA expressions wereassessed using a reverse transcription polymerase chain reaction (RT-PCR and realtimePCR. Furthermore, Oct-4 m RNA expression was assessed using real time PCR.Statistical analysis was performed using ANOVA; and the paired two-sample t test andTukey’s test were used as post-hoc tests for the data analysis of the three sertoli, STOand control cocultures.Results: At the four specified time points, our results showed significant differences (p<0.05in colony numbers and diameters among the sertoli, STO and control groups. The numberand diameter of colonies increased more rapidly in the sertoli coculture than in the othertwo Our results at all four time points also showed significant differences (p<0.05 in themean colony numbers and diameters between the three groups, with the Sertoli coculturehaving the highest mean values for colony numbers and diameters. The RT-PCR results,after two-weeks of culturing, showed that ?1-integrin was expressed in all three groups cocultures,but ?6-integrin was not expressed. Additionally, based on real time PCR results,the three genes (?1-integrin, ?6-integrin, Oct-4 mentioned were also expressed in all threeco cultures groups.Conclusion: Based on the optimal effects of sertoli feeder cells on spermatogonial stemcells in a co culture system, as also confirmed by several other studies, their use is suggestedto achieve better colonization of SSCs.

Seyed Morteza Koruji

2010-01-01

290

Gingival Fibroblasts Display Reduced Adhesion and Spreading on Extracellular Matrix: A Possible Basis for Scarless Tissue Repair?  

OpenAIRE

Unlike skin, oral gingiva do not scar in response to injury. The basis of this difference is likely to be revealed by comparing the responses of dermal and gingival fibroblasts to fibrogenic stimuli. Previously, we showed that, compared to dermal fibroblasts, gingival fibroblasts are less responsive to the potent pro-fibrotic cytokine TGF?, due to a reduced production of endothelin-1 (ET-1). In this report, we show that, compared to dermal fibroblasts, human gingival fibroblasts show reduced...

Guo, Fen; Carter, David E.; Mukhopadhyay, Anuradha; Leask, Andrew

2011-01-01

291

Cytotoxicity of silver dressings on diabetic fibroblasts.  

Science.gov (United States)

A large number of silver-based dressings are commonly used in the management of chronic wounds that are at risk of infection, including diabetic foot ulcers. However, there are still controversies regarding the toxicity of silver dressings on wound healing. The purpose of this study was to objectively test the cytotoxicity of silver dressings on human diabetic fibroblasts. Human diabetic fibroblasts were obtained from the foot skin of four diabetic foot ulcer patients and cultured. The effect of five silver-containing dressing products (Aquacel Ag, Acticoat*Absorbent, Medifoam Ag, Biatain Ag and PolyMem Ag) and their comparable silver-free dressing products on morphology, proliferation and collagen synthesis of the cultured human diabetic fibroblasts were compared in vitro. In addition, extracts of each dressing were tested in order to examine the effect of other chemical components found in the dressings on cytotoxicity. The diabetic fibroblasts cultured with each silver-free dressing adopted the typical dendritic and fusiform shape. On the other hand, the diabetic fibroblasts did not adopt this typical morphology when treated with the different silver dressings. All silver dressings tested in the study reduced the viability of the diabetic fibroblasts and collagen synthesis by 54-70 and 48-68%, respectively, when compared to silver-free dressings. Silver dressings significantly changed the cell morphology and decreased cell proliferation and collagen synthesis of diabetic fibroblasts. Therefore, silver dressings should be used with caution when treating diabetic wounds. PMID:22533495

Zou, Shi-Bo; Yoon, Won-Young; Han, Seung-Kyu; Jeong, Seong-Ho; Cui, Zheng-Jun; Kim, Woo-Kyung

2013-06-01

292

Monensin-induced inhibition of cell spreading in normal and dystrophic human fibroblasts.  

OpenAIRE

Cultured skin fibroblasts from normal individuals and from patients with Duchenne muscular dystrophy spread equally rapidly when seeded on a glass substratum. Exposure to the ionophore monensin substantially suppresses normal and dystrophic fibroblast spreading in serum-free media for up to at least 100 min. Preincubation of normal fibroblasts with monensin causes a further reduction in cell spreading. Dystrophic fibroblasts fail to spread as well as normal cells after monensin preincubation....

Pizzey, J.; Witkowski, J.; Jones, G.

1984-01-01

293

Matched cultures of keratinocytes and fibroblasts derived from normal and NER-deficient mouse models.  

Science.gov (United States)

The uppermost layer of our skin, the epidermis, is formed largely of keratinocytes which constitute the skin's major barrier function and the first line of defence against environmental physical, chemical and biological agents. The subsequent layer, the dermis, which is mainly formed by fibroblasts, has a more supportive function, containing large amounts of collagen, blood vessels and nerve endings and is less directly affected by external insults. Hence it is likely that keratinocytes and fibroblasts have evolved different strategies to cope with the dangers of the environment. Mouse models with various genetic backgrounds in genome care-taking systems, such as DNA repair processes, are well suited to study differences between these two cell types and their implications for cancer and aging. In this chapter we describe a simple procedure to establish long-term keratinocyte and fibroblast cultures from, respectively, the epidermis and dermis of normal or NER-deficient newborn mice. The importance of the external O(2) pressure during the establishment and maintenance of these matched cultures is discussed. PMID:19907995

Pines, Alex; Backendorf, Claude

2010-01-01

294

Skin Diseases: Skin Health and Skin Diseases  

Science.gov (United States)

Skip Navigation Bar Home Current Issue Past Issues Skin Diseases Skin Health and Skin Diseases Past Issues / Fall 2008 Table of Contents ... acne to wrinkles Did you know that your skin is the largest organ of your body? It ...

295

Cytotoxicity and Selectivity in Skin Cancer by SapC-DOPS Nanovesicles  

Directory of Open Access Journals (Sweden)

Full Text Available Squamous cell carcinoma (SCC and melanoma are malignant human cancers of the skin with an annual mortality that exceeds 10,000 cases every year in the USA alone. In this study, the lysosomal protein saposin C (SapC and the phospholipid dioloylphosphatidylserine (DOPS were assembled into cancer-selective nanovesicles (SapC-DOPS and successfully tested using several in vitro and in vivo skin cancer models. Using MTT assay that measures the percentage of cell death, SapC-DOPS cytotoxic effect on three skin tumor cell lines (squamous cell carcinoma, SK-MEL-28, and MeWo was compared to two normal nontumorigenic skin cells lines, normal immortalized keratinocyte (NIK and human fibroblast cell (HFC. We observed that the nanovesicles selectively killed the skin cancer cells by inducing apoptotic cell death whereas untransformed skin cancer cells remained unaffected. Using subcutaneous skin tumor xenografts, animals treated with SapC-DOPS by subcutaneous injection showed a 79.4% by volume tumor reduced compared to the control after 4 days of treatment. We observed that the nanovesicles killed skin cancer cells by inducing apoptotic cell death compared to the control as revealed by TUNEL staining of xenograft tumor sections.

Shadi Abu-Baker

2012-08-01

296

Radiosensitivity in cultured human fibroblasts  

International Nuclear Information System (INIS)

Caution is urged in the use of freshly isolated cultures of human diploid fibroblasts for quantitative studies of radiosensitivity. The distribution of x ray sensitivities of 'normal' human fibroblast cultures of foetal origin (10 subjects, skin or lung biopsy) and post-foetal origin (34 subjects, skin biopsy) are compared with the distribution in 12 patients with ataxia telangiectasia (probability of including any one of these in a normal post-foetal distribution is 0.01%). Cultures from nominally normal subjects showed a broad distribution of D0 range of 98 +- 160 rad and assuming normal distribution, a mean +- one standard deviation of 122 +- 17 rad. Mean D0 values for foetal origin cultures were 117 +- 12; values for post-foetal cultures D0 were 124 +- 18. No systematic variation in D0 was observed for age of donor, number of cell divisions in culture or for cloning efficiency. For ataxia telangiectasia D0 values were 46 +- 7 rad. (U.K.)

297

Hyaluronic Acid and Skin Aging  

Directory of Open Access Journals (Sweden)

Full Text Available Hyaluronic acid (HA, the main and most important constituent of extracellular matrix, is a glycosaminologycan with water-absorbing capacity found in large amount in growing and repairing tissues. One of the main causes of skin problems, particulary in aging skin, is HA deficiency. More than half of the body HA is in the skin and is necessary for the maintenance of internal matrix and several cellular functions. Filler gels containing HA are used to repair skin defects. As these substances are derived from animals and bacteria, not the human, may cause skin reactions and have short half-life. So efforts to maintain and/or increase HA secretion from skin fibroblasts are important in the prevention and treatment of skin aging.

Hossein Abdol Tehrani

2011-09-01

298

Age-related disruption of autophagy in dermal fibroblasts modulates extracellular matrix components  

International Nuclear Information System (INIS)

Highlights: •Autophagosomes accumulate in aged dermal fibroblasts. •Autophagic degradation is impaired in aged dermal fibroblasts. •Autophagy disruption affects extracellular matrix components in dermal fibroblasts. -- Abstract: Autophagy is an intracellular degradative system that is believed to be involved in the aging process. The contribution of autophagy to age-related changes in the human skin is unclear. In this study, we examined the relationship between autophagy and skin aging. Transmission electron microscopy and immunofluorescence microscopy analyses of skin tissue and cultured dermal fibroblasts derived from women of different ages revealed an increase in the number of nascent double-membrane autophagosomes with age. Western blot analysis showed that the amount of LC3-II, a form associated with autophagic vacuolar membranes, was significantly increased in aged dermal fibroblasts compared with that in young dermal fibroblasts. Aged dermal fibroblasts were minimally affected by inhibition of autophagic activity. Although lipofuscin autofluorescence was elevated in aged dermal fibroblasts, the expression of Beclin-1 and Atg5—genes essential for autophagosome formation—was similar between young and aged dermal fibroblasts, suggesting that the increase of autophagosomes in aged dermal fibroblasts was due to impaired autophagic flux rather than an increase in autophagosome formation. Treatment of young dermal fibroblasts with lysosomal protease inhibitors, which mimic the condition of aged dermal fibroblasts with reduced autophagic activity, altered the fibroblast content of type I procollagen, hyaluronan and elastin, and caused a breakdown of collagen fibrils. Collectively, these findings suggest that the autophagy pathway is impaired in aged dermal fibroblasts, which leads to deterioration of dermal integrity and skin fragility

299

Age-related disruption of autophagy in dermal fibroblasts modulates extracellular matrix components  

Energy Technology Data Exchange (ETDEWEB)

Highlights: •Autophagosomes accumulate in aged dermal fibroblasts. •Autophagic degradation is impaired in aged dermal fibroblasts. •Autophagy disruption affects extracellular matrix components in dermal fibroblasts. -- Abstract: Autophagy is an intracellular degradative system that is believed to be involved in the aging process. The contribution of autophagy to age-related changes in the human skin is unclear. In this study, we examined the relationship between autophagy and skin aging. Transmission electron microscopy and immunofluorescence microscopy analyses of skin tissue and cultured dermal fibroblasts derived from women of different ages revealed an increase in the number of nascent double-membrane autophagosomes with age. Western blot analysis showed that the amount of LC3-II, a form associated with autophagic vacuolar membranes, was significantly increased in aged dermal fibroblasts compared with that in young dermal fibroblasts. Aged dermal fibroblasts were minimally affected by inhibition of autophagic activity. Although lipofuscin autofluorescence was elevated in aged dermal fibroblasts, the expression of Beclin-1 and Atg5—genes essential for autophagosome formation—was similar between young and aged dermal fibroblasts, suggesting that the increase of autophagosomes in aged dermal fibroblasts was due to impaired autophagic flux rather than an increase in autophagosome formation. Treatment of young dermal fibroblasts with lysosomal protease inhibitors, which mimic the condition of aged dermal fibroblasts with reduced autophagic activity, altered the fibroblast content of type I procollagen, hyaluronan and elastin, and caused a breakdown of collagen fibrils. Collectively, these findings suggest that the autophagy pathway is impaired in aged dermal fibroblasts, which leads to deterioration of dermal integrity and skin fragility.

Tashiro, Kanae [Skin Research Department, POLA Chemical Industries, Inc., Yokohama (Japan); Division of Pharmaceutical Cell Biology, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka (Japan); Shishido, Mayumi [Skin Research Department, POLA Chemical Industries, Inc., Yokohama (Japan); Fujimoto, Keiko [Division of Pharmaceutical Cell Biology, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka (Japan); Organelle Homeostasis Research Center, Kyushu University, Fukuoka (Japan); Hirota, Yuko [Division of Pharmaceutical Cell Biology, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka (Japan); Yo, Kazuyuki; Gomi, Takamasa [Skin Research Department, POLA Chemical Industries, Inc., Yokohama (Japan); Tanaka, Yoshitaka, E-mail: tanakay@bioc.phar.kyushu-u.ac.jp [Division of Pharmaceutical Cell Biology, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka (Japan); Organelle Homeostasis Research Center, Kyushu University, Fukuoka (Japan)

2014-01-03

300

Pyruvate Dehydrogenase Complex Activity in Normal and Deficient Fibroblasts  

OpenAIRE

Pyruvate dehydrogenase complex (PDC) activity in human skin fibroblasts appears to be regulated by a phosphorylation-dephosphorylation mechanism, as is the case with other animal cells. The enzyme can be activated by pretreating the cells with dichloroacetate (DCA), an inhibitor of pyruvate dehydrogenase kinase, before they are disrupted for measurement of PDC activity. With such treatment, the activity reaches 5-6 nmol/min per mg of protein at 37°C with fibroblasts from infants. Such values...

Sheu, Kwan-fu Rex; Hu, Chii-whei C.; Utter, Merton F.

1981-01-01

301

Fetal ACL Fibroblasts Exhibit Enhanced Cellular Properties Compared with Adults  

OpenAIRE

Fetal tendons and skin heal regeneratively without scar formation. Cells isolated from these fetal tissues exhibit enhanced cellular migration and collagen production in comparison to cells from adult tissue. We determined whether fetal and adult fibroblasts isolated from the anterior cruciate ligament (ACL), a tissue that does not heal regeneratively, exhibit differences in cell migration rates and collagen elaboration. An in vitro migration assay showed fetal ACL fibroblasts migrated twice...

Stalling, Simone S.; Nicoll, Steven B.

2008-01-01

302

Basic fibroblast growth factor induced angiogenesis and prefabricated flap survival.  

Science.gov (United States)

Prefabrication of a latissimus dorsi myocutaneous flap was performed in adult male Landrace pigs. Gelfoam sponges were used as a delivery system for basic fibroblast growth factor (bFGF) at the muscle-subcutaneous tissue interface. Skin survival and angiogenesis were augmented in the growth-factor-treated animals. These data support the use of basic fibroblast growth factor to enhance flap prefabrication. PMID:11316283

Haws, M J; Erdman, D; Bayati, S; Brown, R E; Russell, R C

2001-01-01

303

Adhesion, growth, and matrix production by fibroblasts on laminin substrates  

OpenAIRE

Human embryonic skin fibroblasts have been shown to attach and spread on laminin substrates in the absence of protein synthesis and presence of fibronectin-depleted serum and anti-fibronectin antibodies. Rates of attachment and the type of spreading are virtually identical on fibronectin and laminin-coated substrates with the development of microfilament bundles and focal adhesions. Antibodies to laminin, but not fibronectin, will prevent or reverse fibroblast adhesion...

1983-01-01

304

A comparison of the radiation sensitivities of non-tumorigenic and tumorigenic human hybrid cell lines  

International Nuclear Information System (INIS)

Radiation sensitivities of two related non-tumorigenic and two related tumorigenic human hybrid cell lines (HeLa x skin fibroblast) have been studied. The data show that the transformation from the non-tumorigenic to the tumorigenic state, accompanied by loss of skin fibroblast chromosomes 11 and 14, is not associated with any major changes in radiation sensitivity. The data do indicate, however, a trend toward a steeper and longer initial slope to the cell survival curve for the tumorigenic cell lines, along with a subsequent reduced ability to accumulate sublethal radiation injury at low doses. Both non-tumorigenic and tumorigenic cell lines have the capability of repairing sublethal injury. (author)

305

Modulation of matrix metalloproteinase-7 (matrilysin) secretion in coculture of human colon carcinoma cells with fibroblasts from orthotopic and ectopic organs.  

Science.gov (United States)

Matrix metalloproteinase-7 (MMP-7) is a member of the family of matrix-degrading metalloproteinases that are believed to contribute to the complex process of cancer invasion and metastasis. The secretion level of MMP-7 as assayed by immunoblot analysis was low but distinct in the culture medium of a human colon carcinoma cell line, WIDr, whereas none of the fibroblasts secreted the detectable level of MMP-7. The coculture of WiDr with various human fibroblasts from orthotopic (colon) and ectopic (thyroid, brain, lung, and skin) organs significantly stimulated the secretion of MMP-7 compared with the cultures of individual cells. Reverse transcriptase-polymerase chain reaction analysis and RNA blot analysis suggested that this enhancement occurred at a pretranslational level. The extent of the stimulation was widely varied by the fibroblasts used and was dependent on the cellular ratios and density in the coculture. There may exist a tendency that fibroblasts of orthotopic origin stimulate more extensively than do those of ectopic origin. Moreover, in the coculture of high cell density, normal fibroblasts from the ectopic organs reduced the MMP-7 secretion. The stimulation of MMP-7 secretion may be partially mediated through soluble factor(s); however, direct cell-cell interactions would be required for maximum stimulation. The enhanced MMP-7 secretion was also observed in coculture of colon fibroblasts with other colorectal carcinoma cell lines such as RCM-1 and SW837, which secreted hardly detectable levels of MMP-7 in the individual culture. These results suggest that MMP-7 secretion by colon carcinoma cells is influenced by specific interactions between the carcinoma cells and host fibroblasts. PMID:9220495

Kataoka, H; Meng, J Y; Uchino, H; Nabeshima, K; Kihira, Y; Matuo, Y; Koono, M

1997-01-01

306

Mature Skin  

Science.gov (United States)

... skin Skin of color Stress and skin Sunscreens Tattoos and body piercings Teenage skin Tropical travel Vitamin ... R S T U V W X Y Z Advertising, marketing and sponsorships Legal notice Site map Home ...

307

Dry skin  

Science.gov (United States)

Skin - dry; Winter itch ... Dry skin is common. It happens more often in the winter when cold air outside and heated air inside cause low humidity. Forced-air furnaces make skin even drier. The skin loses moisture and may ...

308

Chemokines and Skin Diseases.  

Science.gov (United States)

Chemokines are small molecules that induce chemotaxis and activation of certain subsets of leukocytes. The expression patterns of chemokines and chemokine receptors are specific to certain organs and cells. Therefore, chemokines are important to elucidate the mechanism of organ-specific human diseases. CCL17 expressed by Langerhans cells, blood endothelial cells, and fibroblasts plays a key role in attracting Th2 cells and tumor cells of adult T-cell leukemia/lymphoma and mycosis fungoides/Sézary syndrome into the skin, developing various Th2-type inflammatory skin diseases as well as cutaneous lymphoma. CCL11 and CCL26 expressed by skin-resident cells, such as fibroblasts, blood endothelial cells, and keratinocytes, induce infiltration of CCR3-expressing cells such as Th2 cells and eosinophils. CCL11 may also serve as an autocrine as well as a paracrine in anaplastic large cell lymphoma. CX3CL1 expressed on blood endothelial cells leads to infiltration of CX3CR1(+) immune cells, such as mast cells, neutrophils, and macrophages, playing important roles in wound healing, tumor immunity, and vasculitis. Biologics targeting chemokines and their receptors are promising strategies for various skin diseases that are resistant to the current therapy. PMID:25182982

Sugaya, Makoto

2014-09-01

309

Elastin peptides activate extracellular signal-regulated kinase 1/2 via a Ras-independent mechanism requiring both p110gamma/Raf-1 and protein kinase A/B-Raf signaling in human skin fibroblasts.  

Science.gov (United States)

Elastin peptides (EPs) produced during cancer progression bind to the elastin binding protein (EBP) found at the surface of dermal fibroblasts, leading to the expression of collagenase-1 gene. The production of this enzyme involved in stromal reaction is caused by the sustained activation of the extracellular signal-regulated kinases 1/2 (ERK1/2) pathway via cAMP/protein kinase A (PKA) and phosphatidylinositol 3-kinase (PI3K). However, the mechanism of these signaling events remains unknown. We show that kappa-elastin (kappaE), a commonly used EP, induces maximum phosphorylation of mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK)1/2 and ERK1/2 after 30 min. The simultaneous inhibition of PKA and PI3K, by N-(2-(p-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide (H89) and 2-(4-morpholynil)-8-phenyl-4H-1-bemzopyran-4-one (LY294002), respectively, blocked MEK1/2 and ERK1/2 phosphorylation, as did lactose, an EBP antagonist. kappaE induced Raf-1 phosphorylation and activation in a PI3K-dependent manner. In our system, the PI3K p110gamma is expressed and activated by betagamma-derived subunits from a pertussis toxin-sensitive G protein after fibroblast stimulation. Pertussis toxin also blocks the Raf-1/MEK1/2/ERK1/2 phosphorylation cascade. In addition, we found that B-Raf is expressed in dermal fibroblasts and activated in a PKA-dependent manner after kappaE treatment, thereby integrating PKA signals to MEK1/2. It is noteworthy that Ras involvement was excluded because ERK1/2 activation by kappaE was not blocked in RasN17-transfected fibroblasts. Together, our results identify a novel Ras-independent ERK1/2 activation system in which p110gamma/Raf-1/MEK1/2 and PKA/B-Raf/MEK1/2 cooperate to activate ERK1/2. Thus, p110gamma and B-Raf seem to be important modulators of dermal fibroblasts physiology and should now qualify as therapeutic targets in strategies aiming at limiting elastin degradation contribution to cancer progression. PMID:15653554

Duca, Laurent; Lambert, Elise; Debret, Romain; Rothhut, Bernard; Blanchevoye, Charlotte; Delacoux, Frédéric; Hornebeck, William; Martiny, Laurent; Debelle, Laurent

2005-04-01

310

DNA hypomethylation and oxidative stress-mediated increase in genomic instability in equine sarcoid-derived fibroblasts.  

Science.gov (United States)

It is widely accepted that equine sarcoid disease, the most common skin associated neoplasm in equids, is induced by bovine papillomavirus (BPV-1). Although BPV-1 DNA has been found in almost all examined sarcoids so far, its detailed impact on the horse's host cell metabolism is largely unknown. We used equine fibroblast cell lines originating from sarcoid biopsies to study BPV-1-associated changes on DNA methylation status and oxidative stress parameters. Sarcoid-derived fibroblasts manifested increased proliferation in vitro, transcriptional rDNA activity (NORs expression) and DNA hypomethylation compared to control cells. Cells isolated from equine sarcoids suffered from oxidative stress: the expression of antioxidant enzymes was decreased and the superoxide production was increased. Moreover, increased ploidy, oxidative DNA damage and micronuclei formation was monitored in sarcoid cells. We postulate that both altered DNA methylation status and redox milieu may affect genomic stability in BPV-1-infected cells and in turn contribute to sarcoid pathology. PMID:22659572

Potocki, Leszek; Lewinska, Anna; Klukowska-Rötzler, Jolanta; Bugno-Poniewierska, Monika; Koch, Christoph; Mählmann, Kathrin; Janda, Josef; Wnuk, Maciej

2012-09-01

311

Lumpy skin disease: Attempted propagation in tick cell lines and presence of viral DNA in field ticks collected from naturally-infected cattle.  

Science.gov (United States)

Lumpy skin disease (LSD) is of substantial economic importance for the cattle industry in Africa and the Near and Middle East. Several insect species are thought to transmit the disease mechanically. Recent transmission studies have demonstrated the first evidence for a role of hard (ixodid) ticks as vectors of lumpy skin disease virus (LSDV). The aim of this study was to attempt in vitro growth of the virus in Rhipicephalus spp. tick cell lines and investigate in vivo the presence of the virus in ticks collected from cattle during LSD outbreaks in Egypt and South Africa. No evidence was obtained for replication of LSDV in tick cell lines although the virus was remarkably stable, remaining viable for 35 days at 28°C in tick cell cultures, in growth medium used for tick cells and in phosphate buffered saline. Viral DNA was detected in two-thirds of the 56 field ticks, making this the first report of the presence of potentially virulent LSDV in ticks collected from naturally infected animals. PMID:25468765

Tuppurainen, E S M; Venter, E H; Coetzer, J A W; Bell-Sakyi, L

2014-11-25

312

Fibroblasts from different sites may promote or inhibit recruitment of flowing lymphocytes by endothelial cells  

OpenAIRE

We examined the hypothesis that stromal fibroblasts modulate the ability of endothelial cells (EC) to recruit lymphocytes in a site-specific manner. PBL were perfused over HUVEC that had been cultured with fibroblasts isolated from the inflamed synovium or the skin of patients with rheumatoid arthritis or osteoarthritis, or from normal synovium, with or without exposure to the inflammatory cytokines TNF-a+IFN-c. Fibroblasts from inflamed synovium, but no others, caused unstimulated HUVEC to b...

Mcgettrick, H. M.; Smith, E.; Filer, A.; Kissane, S.; Salmon, M.; Buckley, Christopher D.; Rainger, G. Ed; Nash, Gerard B.

2009-01-01

313

Endogenous IL-1? from systemic sclerosis fibroblasts induces IL-6 and PDGF-A  

OpenAIRE

It is reported that fibroblasts derived from clinically affected skin areas of patients with systemic sclerosis (SSc) have the ability to overproduce several cytokines and growth factors (i.e., IL-6, PDGF), an ability that might be involved in the pathogenesis of SSc. We have previously shown that the expression of IL-1? was constitutively observed in SSc fibroblasts, whereas this was not detected in normal fibroblasts. Although it was suggested that the aberrant IL-1? production could be a...

Kawaguchi, Yasushi; Hara, Masako; Wright, Timothy M.

1999-01-01

314

The development of a 3D immunocompetent model of human skin  

International Nuclear Information System (INIS)

As the first line of defence, skin is regularly exposed to a variety of biological, physical and chemical insults. Therefore, determining the skin sensitization potential of new chemicals is of paramount importance from the safety assessment and regulatory point of view. Given the questionable biological relevance of animal models to human as well as ethical and regulatory pressure to limit or stop the use of animal models for safety testing, there is a need for developing simple yet physiologically relevant models of human skin. Herein, we describe the construction of a novel immunocompetent 3D human skin model comprising of dendritic cells co-cultured with keratinocytes and fibroblasts. This model culture system is simple to assemble with readily-available components and importantly, can be separated into its constitutive individual layers to allow further insight into cell–cell interactions and detailed studies of the mechanisms of skin sensitization. In this study, using non-degradable microfibre scaffolds and a cell-laden gel, we have engineered a multilayer 3D immunocompetent model comprised of keratinocytes and fibroblasts that are interspersed with dendritic cells. We have characterized this model using a combination of confocal microscopy, immuno-histochemistry and scanning electron microscopy and have shown differentiation of the epidermal layer and formation of an epidermal barrier. Crucially the immune cells in the model are able to migrate and remain re model are able to migrate and remain responsive to stimulation with skin sensitizers even at low concentrations. We therefore suggest this new biologically relevant skin model will prove valuable in investigating the mechanisms of allergic contact dermatitis and other skin pathologies in human. Once fully optimized, this model can also be used as a platform for testing the allergenic potential of new chemicals and drug leads. (paper)

315

Zeaxanthin inhibits PDGF-BB-induced migration in human dermal fibroblasts.  

Science.gov (United States)

Zeaxanthin is the dihydroxy carotenoid and is distributed in our daily foods. Various natural carotenoids, including zeaxanthin, have been shown to inhibit proliferation of several types of cancer cells, but available data on the effect of zeaxanthin on skin fibroblasts and melanoma cells are limited. Platelet-derived growth factor (PDGF) functions as a chemotactic factor for dermal fibroblasts and plays an important role in the progression of melanoma. In this study, we investigated the effects of zeaxanthin on the migration of skin fibroblasts induced by PDGF-BB and melanoma cells. We demonstrated that zeaxanthin inhibited PDGF-BB-induced skin fibroblast migration on collagen and gelatin by a modified Boyden chamber system. The electric cell-substrate impedance sensing (ECIS) method also showed similar inhibitory effects of zeaxanthin on the migration of fibroblasts. In functional studies, zeaxanthin decreased melanoma-induced fibroblast migration in a non-contact coculture system and also the migration stimulated by melanoma-derived conditioned medium. Further analysis showed that zeaxanthin attenuated PDGF-BB and melanoma-conditioned medium induced phosphorylation of PDGFR-beta and MAP kinase in a concentration-dependent manner in human skin fibroblasts. However, these effects did not result from direct interaction of zeaxanthin with PDGF-BB. Thus, our results provide the first evidence showing that zeaxanthin is an effective inhibitor of migration of stromal fibroblasts induced by PDGF-BB and melanoma cells and this effect may further support its antitumor potential. PMID:20482615

Wu, Nan-Lin; Chiang, Yuh-Chiau; Huang, Chieh-Chen; Fang, Jia-You; Chen, Der-Fang; Hung, Chi-Feng

2010-08-01

316

Heterogeneity in the radiation survival curves and biochemical properties of human lung cancer cell lines  

International Nuclear Information System (INIS)

Human lung cancers of distinct histology exhibit different responses to radiation therapy in vivo. For examination of the basis of this phenomenon, the radiation survival curves and levels of relevant enzymes were determined in 16 lung cancer cell lines derived from tumors of different histology. These included lines from 5 adenocarcinomas, 7 small cell tumors, 3 variant small cell tumors, and 1 large cell tumor. These findings were compared to those obtained with the use of a normal skin fibroblast cell line. Whether cloned in liquid culture or soft agarose, cell lines had similar radiation survival curves. These curves were consistent with the apparent in vivo radiation responsiveness of the tumors. Although considerable heterogeneity in radiation survival curves was observed among the cell lines, cells from large cell lines and small variant lines had pronounced shoulders and extrapolation numbers (n) from 5.6 to 14. In contrast, cells from small cell lines and adenocarcinoma cell lines were more sensitive (-n values of 1-3.3). In these cell lines, levels of DNA polymerase beta, glutathione (GSH), GSH transferase, GSH reductase (NAD(P)H), gamma-glutamyltransferase did not correlate with radiation parameters of sensitivity. DNA polymerase beta and GSH levels were, however, higher than those in a line of normal skin fibroblasts. These cell lines may be useful in identifying the basis of the variable responsiveness of human lung cancer cells to ionizing radiationng cancer cells to ionizing radiation

317

Polypeptide structure of a human dermal fibroblast-activating factor (FAF) derived from the U937 cultured line of human monocyte-like cells  

International Nuclear Information System (INIS)

Six liter batches of 1 x 106 U937 cells/ml of serum-free RPMI medium were incubated with 100 ng/ml of phorbol myristate acetate for 48 hr at 370C in 5% CO2 in air to generate FAFs, as quantified by the stimulation of uptake of [3H]thymidine by quiescent human dermal fibroblasts. Filtration of the supernatants on Sephadex G-75 resolved two FAFs of approximately 40,000 and 10-13,000 daltons. The latter principle was purified to homogeneity by sequential Sephadex G-50 filtration, revealing an apparent m.w. of 7-8000, Mono-Q FPLC anion-exchange chromatography with a linear gradient from 20 mM Tris-HCl (pH 8.3) to 0.5 M NaCl-20 mM Tris-HCl in 30 min, and two cycles of high-performance liquid chromatography (HPLC) on a 300 A pore 10 ?m C4 column at 1 ml/min with 0.05% trifluoroacetic acid (TFA) in water to 30:70 (v:v) and then to 60:40 (v:v) acetonitrile: 0.05% TFA linearly in 15 min and 30 min, respectively, The FAF activity eluted from HPLC in a sharp peak of O.D. 215 nm at 45% acetonitrile. Analyses of amino acid composition of the highly purified 7-8000 dalton FAF-U937 revealed 37% hydrophobic, 14% basic, and 21% acidic or amide residues, as well as one tryrosine and one methionine. This U937 cell-derived FAF appears to be a unique acidic polypeptide growth factor

318

Increased biosynthesis and processing of fibronectin in fibroblasts from diabetic mice.  

OpenAIRE

Diabetic connective tissues exhibit a deranged regulation of extracellular matrix biosynthesis. Fibronectin is shown to be increased in human dermal connective tissue by immunofluorescence, mainly at the dermoepidermal and capillary basement membranes. The rate of fibronectin biosynthesis, excretion, and incorporation in a pericellular polymeric form was investigated using genetically diabetic KK mouse skin and fibroblasts as compared to Swiss and C57BL mouse skin and fibroblasts. The rate of...

Phan-thanh, L.; Robert, L.; Derouette, J. C.; Labat-robert, J.

1987-01-01

319

Dexamethasone prevents granulocyte-macrophage colony-stimulating factor-induced nuclear factor-kappaB activation, inducible nitric oxide synthase expression and nitric oxide production in a skin dendritic cell line.  

OpenAIRE

AIMS: Nitric oxide (NO) has been increasingly implicated in inflammatory skin diseases, namely in allergic contact dermatitis. In this work, we investigated the effect of dexamethasone on NO production induced by the epidermal cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) in a mouse fetal skin dendritic cell line. METHODS: NO production was assessed by the method of Griess. Expression of the inducible isoform of nitric oxide synthase (iNOS) protein was evaluated by wester...

Vital, Al; Gonc?alo, Margarida; Cruz, Mt; Figueiredo, A.; Duarte, Cb; Lopes, Mc

2003-01-01

320

Constitutive production of inflammatory and mitogenic cytokines by rheumatoid synovial fibroblasts  

OpenAIRE

Conditioned media obtained from fibroblasts cultured from rheumatoid and certain other inflammatory synovia were observed to stimulate [3H]thymidine incorporation in an indicator murine fibroblast line. Synovial fibroblasts derived from the joints of patients with osteoarthritis did not display this property. This effect persisted in culture for many weeks and occurred in the absence of co-stimulatory immune cells. Antibody neutralization studies implicated a role for basic fibroblast growth ...

1991-01-01

321

DNA repair in human fibroblasts, as reflected by host-cell reactivation of a transfected UV-irradiated luciferase gene, is not related to donor age  

International Nuclear Information System (INIS)

The effect of donor age on the ability of mammalian cells to repair ultraviolet (UV)-induced DNA damage has been studied using several approaches, most recently via assays that measure the host-cell reactivation (HCR) of UV-irradiated reporter gene-containing plasmid vectors following their transfection into cells. Plasmid HCR assays indirectly quantify a cell line's ability to perform nucleotide excision repair (NER) by measuring the enzyme activity of the repaired reporter gene, e.g., chloramphenical acetyltransferase (cat) or luciferase (luc), and are useful in studies investigating whether increasing age may be a risk factor for the deficient repair of potentially cancer-causing, sunlight-induced, DNA lesions in skin cells. In our study, we quantified the DNA repair ability of cultured, nontransformed, human skin fibroblast lines through their HCR of a transfected UV-C-irradiated plasmid containing luc. HCR was measured at various times after transfection in five lines from normal donors of ages 21-96 years, and from one donor who had xeroderma pigmentosum (XP). The normal lines displayed increasing HCR at successive post-transfection time points and showed no significant correlation between HCR and donor age. The XP-A line, known to be markedly deficient in NER of UV-induced DNA damage, showed minimal evidence of HCR compared to the normal lines. To further assess potential variation in HCR with donor age, fibroblast lines from five old donors, ages 84-94 years, s from five old donors, ages 84-94 years, were compared with lines from five young donors, ages 17-26 years. While significant differences in HCR were found between some lines, no significant difference was found between the young and old age groups (P=0.44). Our study provides no indication that the higher incidence of skin cancer observed with increasing age is due to an age-related decrease in the ability to repair UV-induced DNA damage

322

Loss of PPAR? expression by fibroblasts enhances dermal wound closure  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Peroxisome proliferator-activated receptor (PPAR? may be a key regulator of connective tissue deposition and remodeling in vivo. PPAR? expression is reduced in dermal fibroblasts isolated from fibrotic areas of scleroderma patients; PPAR? agonists suppress the persistent fibrotic phenotype of this cell type. Previously, we showed that loss of PPAR? expression in fibroblasts resulted in enhanced bleomycin-induced skin fibrosis. However, whether loss of PPAR? expression in skin fibroblasts affects cutaneous tissue repair or homeostasis is unknown. Results Mice deleted for PPAR? in skin fibroblasts show an enhanced rate of dermal wound closure, concomitant with elevated phosphorylation of Smad3, Akt and ERK, and increased expression of proliferating cell nuclear antigen (PCNA, collagen, ?-smooth muscle actin (?-SMA and CCN2. Conversely, dermal homeostasis was not appreciably affected by loss of PPAR? expression. Conclusion PPAR? expression by fibroblasts suppresses cutaneous tissue repair. In the future, direct PPAR? antagonists and agonists might be of clinical benefit in controlling chronic wounds or scarring, respectively.

Sha Wei

2012-04-01

323

Development of a human mitochondrial oligonucleotide microarray (h-MitoArray and gene expression analysis of fibroblast cell lines from 13 patients with isolated F1Fo ATP synthase deficiency  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background To strengthen research and differential diagnostics of mitochondrial disorders, we constructed and validated an oligonucleotide microarray (h-MitoArray allowing expression analysis of 1632 human genes involved in mitochondrial biology, cell cycle regulation, signal transduction and apoptosis. Using h-MitoArray we analyzed gene expression profiles in 9 control and 13 fibroblast cell lines from patients with F1Fo ATP synthase deficiency consisting of 2 patients with mt9205?TA microdeletion and a genetically heterogeneous group of 11 patients with not yet characterized nuclear defects. Analysing gene expression profiles, we attempted to classify patients into expected defect specific subgroups, and subsequently reveal group specific compensatory changes, identify potential phenotype causing pathways and define candidate disease causing genes. Results Molecular studies, in combination with unsupervised clustering methods, defined three subgroups of patient cell lines – M group with mtDNA mutation and N1 and N2 groups with nuclear defect. Comparison of expression profiles and functional annotation, gene enrichment and pathway analyses of differentially expressed genes revealed in the M group a transcription profile suggestive of synchronized suppression of mitochondrial biogenesis and G1/S arrest. The N1 group showed elevated expression of complex I and reduced expression of complexes III, V, and V-type ATP synthase subunit genes, reduced expression of genes involved in phosphorylation dependent signaling along MAPK, Jak-STAT, JNK, and p38 MAP kinase pathways, signs of activated apoptosis and oxidative stress resembling phenotype of premature senescent fibroblasts. No specific functionally meaningful changes, except of signs of activated apoptosis, were detected in the N2 group. Evaluation of individual gene expression profiles confirmed already known ATP6/ATP8 defect in patients from the M group and indicated several candidate disease causing genes for nuclear defects. Conclusion Our analysis showed that deficiency in the ATP synthase protein complex amount is generally accompanied by only minor changes in expression of ATP synthase related genes. It also suggested that the site (mtDNA vs nuclear DNA and the severity (ATP synthase content of the underlying defect have diverse effects on cellular gene expression phenotypes, which warrants further investigation of cell cycle regulatory and signal transduction pathways in other OXPHOS disorders and related pharmacological models.

Hansíková Hana

2008-01-01

324

Sagging Skin  

Science.gov (United States)

... for Every Season How to Choose the Best Skin Care Products In This Section Dermatologic Surgery What is dermatologic ... for Every Season How to Choose the Best Skin Care Products Sagging Skin Treatment Options Learn more about the ...

325

Skin Conditions  

Science.gov (United States)

Your skin is your body's largest organ. It covers and protects your body. Your skin Holds body fluids in, preventing dehydration Keeps harmful ... it Anything that irritates, clogs, or inflames your skin can cause symptoms such as redness, swelling, burning, ...

326

Establishment and Biological Characteristics of Hereford Cattle Fibroblast Bank  

OpenAIRE

A fibroblast line from kindey tissue of Hereford cattle was established successfully by direct culture of explants and biology cryopreservation techniques. The cell line contained 101 tubes of frozen cells from 34 primary kidney samples. Biological analysis showed that the cells were morphologically consistent with fibroblasts and the growth curve was sigmoidal with a Population Doubling Time (PDT) of 35 h.The average viability of the cells was 95.8% before freezing and 93.4% after thawing. C...

Weijun Guan; Mei Li; Changli Li; Wenxiu Zhang; Shen Wu; Yuehui Ma

2012-01-01

327

Doubling potential of fibroblasts from different species after ionising radiation  

International Nuclear Information System (INIS)

It is stated that whereas chicken fibroblasts invariably die after a certain number of doublings in vitro, and this fact is never altered by chemical or physical agents, mouse fibroblasts invariably acquire spontaneously an infinite growth potential. In the human species fibroblasts never acquire spontaneously the capacity to divide for ever, although they can become permanent cell lines after treatment with certain viruses. This behaviour of fibroblasts in vitro has been attributed to different nutritional requirements. Experiments are described with human and mouse fibroblasts in which it was found that the response to ionising radiation matches the relative tendencies of the fibroblasts to yield permanent cell lines. Irradiation was commenced during the phase of active proliferation. Human fibroblast cultures irradiated with 100 R stopped dividing earlier than the controls, whereas cultures irradiated with 200, 300 and 500 R had the same lifespan as the control cultures. Cultures irradiated with 400 R showed the longest survival. With mouse fibroblasts the growth curves of the irradiated cells were of the same type as in the controls, but recovery occurred earlier. The results indicated that ionising radiation accelerates a natural phenomenon; in cells with a limited growth potential (chicken) it shortens the lifespan, whereas in cells that can acquire an unlimited growth potential (mouse) it accelerates acquisition of this potential; human fibroblasts showed an in potential; human fibroblasts showed an intermediate response, since ionising radiation neither established the cultures as with mouse cells nor reduced the number of cells produced as with chicken fibroblasts. Possible explanations for the different behaviour of the species are offered. (U.K.)

328

Multiple RNAs expressed from the int-2 gene in mouse embryonal carcinoma cell lines encode a protein with homology to fibroblast growth factors.  

OpenAIRE

Mouse embryonal carcinoma cell lines that differ in their patterns of expression of the potential oncogene int-2 have been exploited in a structural analysis of the multiple RNA transcripts characteristic of this gene. Ribonuclease protection experiments indicate that four major classes of int-2 RNA initiate at heterogeneous cap sites within two distinct promoter regions, P1 and P2, spanning approximately 50 and 150 bp respectively. The more downstream promoter P2 is located in a region of th...

Smith, R.; Peters, G.; Dickson, C.

1988-01-01

329

Fibroblast differentiation in subcutaneous fibrosis after postmastectomy radiotherapy  

Energy Technology Data Exchange (ETDEWEB)

In order to acquire a better understanding of the mechanism of radiation-induced fibrosis, we studied the differentiation of normal skin fibroblasts cultured from breast cancer radiotherapy patients with different risk of fibrosis. The differentiation state of fibroblasts was characterized in clonal cultures using established cytomorphological criteria. Collagen synthesis was determined by 3H-proline incorporation into pepsin-resistant protein. Radiation-induced inactivation of fibroblasts was paralleled by an increase in terminally differentiated fibrocytes, demonstrating that premature terminal differentiation is an important response to irradiation of fibroblasts from radiotherapy patients. Surviving colony-forming fibroblasts showed a change in differentiation with an increase in the ratio L:E of progenitor fibroblasts in late (L) compared to early (E) differentiation states. Furthermore, increased collagen production was observed after irradiation. The results provide evidence supporting a role of terminal fibroblast differentiation in radiation-induced fibrosis and imply that the progenitor population surviving radiotherapy might be more prone to terminal differentiation than before radiotherapy.

Herskind, C.; Johansen, J.; Bentzen, S.M.; Overgaard, M.; Overgaard, J.; Bamberg, M.; Rodemann, H.P. [Univ. of Tuebingen (Germany). Section of Radiobiology and Molecular Environmental Research

2000-07-01

330

Fibroblast differentiation in subcutaneous fibrosis after postmastectomy radiotherapy  

DEFF Research Database (Denmark)

In order to acquire a better understanding of the mechanism of radiation-induced fibrosis, we studied the differentiation of normal skin fibroblasts cultured from breast cancer radiotherapy patients with different risk of fibrosis. The differentiation state of fibroblasts was characterized in clonal cultures using established cytomorphological criteria. Collagen synthesis was determined by 3H-proline incorporation into pepsin-resistant protein. Radiation-induced inactivation of fibroblasts was paralleled by an increase in terminally differentiated fibrocytes, demonstrating that premature terminal differentiation is an important response to irradiation of fibroblasts from radiotherapy patients. Surviving colony-forming fibroblasts showed a change in differentiation with an increase in the ratio L:E of progenitor fibroblasts in late (L) compared to early (E) differentiation states. Furthermore, increased collagen production was observed after irradiation. The results provide evidence supporting a role of terminal fibroblast differentiation in radiation-induced fibrosis and imply that the progenitor population surviving radiotherapy might be more prone to terminal differentiation than before radiotherapy.

Herskind, C; Johansen, J

2000-01-01

331

Abscisic acid ameliorates the systemic sclerosis fibroblast phenotype in vitro  

International Nuclear Information System (INIS)

Highlights: ? ABA is an endogenous hormone in humans, regulating different cell responses. ? ABA reverts some of the functions altered in SSc fibroblasts to a normal phenotype. ? UV-B irradiation increases ABA content in SSc cultures. ? SSc fibroblasts could benefit from exposure to ABA and/or to UV-B. -- Abstract: The phytohormone abscisic acid (ABA) has been recently identified as an endogenous hormone in humans, regulating different cell functions, including inflammatory processes, insulin release and glucose uptake. Systemic sclerosis (SSc) is a chronic inflammatory disease resulting in fibrosis of skin and internal organs. In this study, we investigated the effect of exogenous ABA on fibroblasts obtained from healthy subjects and from SSc patients. Migration of control fibroblasts induced by ABA was comparable to that induced by transforming growth factor-? (TGF-?). Conversely, migration toward ABA, but not toward TGF-?, was impaired in SSc fibroblasts. In addition, ABA increased cell proliferation in fibroblasts from SSc patients, but not from healthy subjects. Most importantly, presence of ABA significantly decreased collagen deposition by SSc fibroblasts, at the same time increasing matrix metalloproteinase-1 activity and decreasing the expression level of tissue inhibitor of metalloproteinase (TIMP-1). Thus, exogenously added ABA appeared to revert some of the functions altered in SSc fibroblasts to a normal phenotype. Interestingly, ABA levels in plasma from SSc patients were found to be significantly lower than in healthy subjects. UV-B irradiation induced an almost 3-fold increase in ABA content in SSc cultures. Altogether, these results suggest that the fibrotic skin lesions in SSc patients could benefit from exposure to high(er) ABA levels.

332

Fibroblasts induce heparin synthesis in chondroitin sulfate E containing human bone marrow-derived mast cells  

International Nuclear Information System (INIS)

Human bone marrow-derived mast cells (hBMMCs), differentiated in vitro in suspension culture and under the influence of human peripheral blood mononuclear cells conditioned medium (hCM), were tested for their response to recombinant human interleukin-3 (rhIL-3) and for their behavior in different microenvironments. The hBMMCs were incubated in the presence of rhIL-3 and the changes in their proliferation rate were determined. Recombinant hIL-3 induced a more than sixfold increase in 3H-thymidine uptake into the hBMMC DNA in a dose-dependent manner. Human CM used as a control for proliferation response induced a more than eightfold maximal proliferation rate increase. Rabbit anti-rhIL-3 completely inhibited hBMMC 3H-thymidine uptake induced by rhIL-3 and decreased the hCM-induced proliferation by approximately 50%. These hBMMCs were cocultured with four different mytomicin C-treated cell monolayers and assayed for phenotypic changes. After only 2 days in coculture with either embryonic mouse skin-derived fibroblasts (MESFs) or human skin-derived fibroblasts (HSFs), a marked increase in granule number and density was noted on staining with toluidine blue. Mast cells that initially stained alcian blue+/safranin- at day 0 of coculture became alcian blue+/safranin+ during the coculture period. Human BMMC proteoglycan synthesis shifted from approximately 85% chondroitin sulfate E to approximately 60% heparin within 14 to 19 days of coculture with the MESF monolayer and to acoculture with the MESF monolayer and to approximately 50% heparin within 19 days of coculture with the HSF monolayer. None of the above-mentioned changes were noted in cocultures of hBMMCs with 3T3 cell line fibroblast monolayers or in cocultures with bovine vascular endothelium (BVE) cell monolayers

333

How to Approach Finnish Retail Market when Launching a New Skin Care Line: a Case Study of Créations Couleurs  

OpenAIRE

The cosmetics industry is one of the biggest lines of businesses in the world. In Finland people spend thousands of Euros per year on cosmetic and hygiene products. Everything changes constantly and this has reflected to the cosmetics industry as well as consumers. People increasingly desire several options to choose from and want quick results. The topic for this thesis came from a French cosmetic company Créations Couleurs which develops and manufactures raw materials for different cos...

Nordenswan, Katarina; Huttunen, Anne

2012-01-01

334

Laser printing of skin cells and human stem cells.  

Science.gov (United States)

Laser printing based on laser-induced forward transfer (LIFT) is a new biofabrication technique for the arrangement of biological materials or living cells in well-defined patterns. In the current study, skin cell lines (fibroblasts/keratinocytes) and human mesenchymal stem cells (hMSC) were chosen for laser printing experiments due to their high potential in regeneration of human skin and new application possibilities of stem cell therapy. To evaluate the influence of LIFT on the cells, their survival rate, their proliferation and apoptotic activity, and the DNA damages and modifications of their cell surface markers were assessed and statistically evaluated over several days. The cells survived the transfer procedure with a rate of 98%? +/- 1% standard error of the mean (skin cells) and 90%? +/- 10% (hMSC), respectively. All used cell types maintain their ability to proliferate after LIFT. Further, skin cells and hMSC did not show an increase of apoptosis or DNA fragmentation. In addition, the hMSC keep their phenotype as proven by fluorescence activated cell sorting (FACS) analysis. This study demonstrates LIFT as a suitable technique for unharmed computer-controlled positioning of different cell types and a promising tool for future applications in the ex vivo generation of tissue replacements. PMID:19883209

Koch, Lothar; Kuhn, Stefanie; Sorg, Heiko; Gruene, Martin; Schlie, Sabrina; Gaebel, Ralf; Polchow, Bianca; Reimers, Kerstin; Stoelting, Stephanie; Ma, Nan; Vogt, Peter M; Steinhoff, Gustav; Chichkov, Boris

2010-10-01

335

Skin Cancer  

Science.gov (United States)

... Media resources Stats and facts Conditions Skin cancer Skin cancer Incidence rates More than 3.5 million nonmelanoma skin cancers ... 62 percent and 16 percent, respectively. 2 Mortality rates Approximately 75 percent of skin cancer deaths are from melanoma. 2 On average, one ...

336

Radiation response in vitro of fibroblasts from a Fanconi anemia patient with marked clinical radiosensitivity  

Energy Technology Data Exchange (ETDEWEB)

Background: fanconi anemia (FA) is an autosomal recessive chromosome instability disorder characterized by progressive pancytopenia and cancer susceptibility. The risks of radiation therapy in FA patients who have cancer remain to be investigated. Recently, Marcou et al. (2001) reported a case of severe clinical radiosensitivity in a female FA patient with a tonsillar squamous cell carcinoma treated by radiotherapy. By contrast, her in vitro irradiated skin fibroblasts revealed nearly normal radiosensitivity as determined by the colony survival assay. Material and methods: in view of this discrepancy, the radiation response of this particular FA fibroblast strain (designated 425BR) was further analyzed in the present study by means of the alkaline single-cell gel electrophoresis (Comet) assay, and also by the cytochalasin-blocked micronuclei (MN) test. In addition, the expression levels of DNA repair proteins, hMre11, Rad50, and Rad51, were investigated using Western blot and foci immunofluorescence staining. Results: the Comet assay revealed that the initial DNA fragmentation in irradiated FA cells was two times higher and the DNA rejoining process was three times slower than that in control (1BR3) fibroblasts. Moreover, although the baseline level of MNs was lower in FA cells than in controls, the FA fibroblasts were more prone (about two times) to MN production than control cells when irradiated with 2-4 Gy. Western blot analysis of the DNA repair proteins (hMre11, Rad50, and Rad51) did not reveal any abnormalities in protein expression levels or their migration patterns in the fibroblasts derived from an FA patient either before or after irradiation. At the same time, in vitro irradiated cells from the FA patient exhibited a significantly reduced number of nuclei with focally concentrated DNA repair Rad51 protein than in control cells. Conclusion: the increased DNA damage and MN induction in irradiated FA fibroblasts, and the reduction of the formation of DNA repair foci containing Rad51 suggest a possible link to the profound clinical radiosensitivity reported earlier for this FA patient. The findings on this particular FA cell strain presented in the study point toward the difficulties involved in the prediction of the radiation response of cell lines and tumors based solely on the colony survival test. (orig.)

Djuzenova, C.; Flentje, M. [Dept. of Radiotherapy, Univ. of Wuerzburg, Wuerzburg (Germany); Plowman, P.N. [Radiotherapy/Clinical Oncology, St. Bartholomew' s Hospital, London (United Kingdom)

2004-12-01

337

Radiation response in vitro of fibroblasts from a Fanconi anemia patient with marked clinical radiosensitivity  

International Nuclear Information System (INIS)

Background: fanconi anemia (FA) is an autosomal recessive chromosome instability disorder characterized by progressive pancytopenia and cancer susceptibility. The risks of radiation therapy in FA patients who have cancer remain to be investigated. Recently, Marcou et al. (2001) reported a case of severe clinical radiosensitivity in a female FA patient with a tonsillar squamous cell carcinoma treated by radiotherapy. By contrast, her in vitro irradiated skin fibroblasts revealed nearly normal radiosensitivity as determined by the colony survival assay. Material and methods: in view of this discrepancy, the radiation response of this particular FA fibroblast strain (designated 425BR) was further analyzed in the present study by means of the alkaline single-cell gel electrophoresis (Comet) assay, and also by the cytochalasin-blocked micronuclei (MN) test. In addition, the expression levels of DNA repair proteins, hMre11, Rad50, and Rad51, were investigated using Western blot and foci immunofluorescence staining. Results: the Comet assay revealed that the initial DNA fragmentation in irradiated FA cells was two times higher and the DNA rejoining process was three times slower than that in control (1BR3) fibroblasts. Moreover, although the baseline level of MNs was lower in FA cells than in controls, the FA fibroblasts were more prone (about two times) to MN production than control cells when irradiated with 2-4 Gy. Western blot analysis of the DNA repair proteins (hMre11,alysis of the DNA repair proteins (hMre11, Rad50, and Rad51) did not reveal any abnormalities in protein expression levels or their migration patterns in the fibroblasts derived from an FA patient either before or after irradiation. At the same time, in vitro irradiated cells from the FA patient exhibited a significantly reduced number of nuclei with focally concentrated DNA repair Rad51 protein than in control cells. Conclusion: the increased DNA damage and MN induction in irradiated FA fibroblasts, and the reduction of the formation of DNA repair foci containing Rad51 suggest a possible link to the profound clinical radiosensitivity reported earlier for this FA patient. The findings on this particular FA cell strain presented in the study point toward the difficulties involved in the prediction of the radiation response of cell lines and tumors based solely on the colony survival test. (orig.)

338

Evaluation of cytotoxic activities of snake venoms toward breast (MCF-7) and skin cancer (A-375) cell lines.  

Science.gov (United States)

Snake venoms are mixtures of bioactive proteins and peptides that exhibit diverse biochemical activities. This wide array of pharmacologies associated with snake venoms has made them attractive sources for research into potentially novel therapeutics, and several venom-derived drugs are now in use. In the current study we performed a broad screen of a variety of venoms (61 taxa) from the major venomous snake families (Viperidae, Elapidae and "Colubridae") in order to examine cytotoxic effects toward MCF-7 breast cancer cells and A-375 melanoma cells. MTT cell viability assays of cancer cells incubated with crude venoms revealed that most venoms showed significant cytotoxicity. We further investigated venom from the Red-bellied Blacksnake (Pseudechis porphyriacus); venom was fractionated by ion exchange fast protein liquid chromatography and several cytotoxic components were isolated. SDS-PAGE and MALDI-TOF mass spectrometry were used to identify the compounds in this venom responsible for the cytotoxic effects. In general, viper venoms were potently cytotoxic, with MCF-7 cells showing greater sensitivity, while elapid and colubrid venoms were much less toxic; notable exceptions included the elapid genera Micrurus, Naja and Pseudechis, which were quite cytotoxic to both cell lines. However, venoms with the most potent cytotoxicity were often not those with low mouse LD50s, including some dangerously venomous viperids and Australian elapids. This study confirmed that many venoms contain cytotoxic compounds, including catalytic PLA2s, and several venoms also showed significant differential toxicity toward the two cancer cell lines. Our results indicate that several previously uncharacterized venoms could contain promising lead compounds for drug development. PMID:25407733

Bradshaw, Michael J; Saviola, Anthony J; Fesler, Elizabeth; Mackessy, Stephen P

2014-11-19

339

Induction of chromosomal aberrations in human primary fibroblasts and immortalized cancer cells exposed to extremely-low-frequency electromagnetic fields  

International Nuclear Information System (INIS)

Rapidly increasing possibilities of exposure to environmental extremely low-frequency electromagnetic fields have become a topic of worldwide investigation. Epidemiological and laboratory studies suggest that exposure to extremely low-frequency electromagnetic fields may increase cancer risk therefore assessment of chromosomal damage in various cell lines might be of predictive value for future risk estimation. Materials and Methods: Primary cultures of fibroblasts from human skin biopsy were exposed to continuous extremely low-frequency electromagnetic fields (3, 50 and 60 Hz, sinusoidal, 3h, and 4 m T). Also immortalized cell lines, SW480, MCF-7 and 1321N1 were exposed to continuous extremely low-frequency electromagnetic fields (50 Hz, sinusoidal, 3 h, 4 m T). Metaphase plates Were prepared according to standard methods and stained in 5% Giemsa solution. Chromosomal aberrations of both chromosome and chromatid types were scored to evaluate the effects of extremely low-frequency electromagnetic fields on primary or established cell lines. Results: Results indicate that by increasing the frequency of extremely low-frequency electromagnetic fields, chromosomal aberrations were increased up to 7-fold above background levels in primary human fibroblast cells. In addition, continuous exposure to a 50 Hz electromagnetic field led to a significant increase in chromosomal aberrations in SW480, MCF-7 and 1321N1 cell lines compared to sham control. Conclusion: Results obtaine sham control. Conclusion: Results obtained indicate that extremely low-frequency electromagnetic fields has the potential for induction of chromosomal aberrations in all cell types.

340

Chloride transport in human fibroblasts is activated by hypotonic shock  

Energy Technology Data Exchange (ETDEWEB)

Incubation of human skin fibroblasts in hypotonic media induced the activation of {sup 36}Cl- efflux which was roughly proportional to the decrease in the osmolality of the media. The efflux of {sup 36}Cl- was insensitive to DIDS plus furosemide and inhibited by addition of a Cl- channel blocker such as 5-nitro-2-(3-phenyl propylamino) benzoic acid (NPPB). We propose that a conductive pathway for Cl- transport, almost silent in isotonic conditions, is activated by exposing human fibroblasts to hypotonic shock, this conclusion being supported by evidence that also {sup 36}Cl- influx was enhanced by hypotonic medium.

Rugolo, M.; Mastocola, T.; Flamigni, A.; Lenaz, G. (Universita' di Bologna (Italy))

1989-05-15

341

Primary cell culture and morphological characterization of canine dermal papilla cells and dermal fibroblasts.  

Science.gov (United States)

Skin biopsies were taken from female dogs, the primary hair follicles isolated and the dermal papilla dissected. After incubation in supplemented Amniomax complete C100 medium in 24-well culture plates, the dermal papilla cells (DPC) grew to confluence within 3 weeks. Thereafter, they were subcultivated every 7 days. Dermal fibroblast (DFB) cultures were established by explant culture of interfollicular dermis in serum-free medium, where they reached confluence in 10 days. They were subcultivated every 5 days. For immunohistochemistry, cells were grown on cover slips for 24 h, fixed and stained with antibodies against collagen IV and laminin. DPC showed an aggregative growth pattern and formation of pseudopapillae. Intensive staining for collagen IV and laminin could be observed until the sixth passage. DFB grew as branching, parallel lines and showed only weak staining for collagen IV and laminin. PMID:11896964

Bratka-Robia, Christine B; Mitteregger, Gerda; Aichinger, Amanda; Egerbacher, Monika; Helmreich, Magdalena; Bamberg, Elmar

2002-02-01

342

Development and evaluation of a skin organ model for the analysis of radiation effects  

Energy Technology Data Exchange (ETDEWEB)

Background and purpose: the reaction of tissues to ionizing radiation involves alterations in cell-cell and cell-matrix interactions mediated by cellular adhesion molecules. The aim of this study was to develop and evaluate an artificial skin organ model for the analysis of radiation effects. Material and methods: a human co-culture system consisting of the spontaneously immortalized keratinocyte cell line HaCaT and primary HDFa fibroblasts embedded into a collagen sponge was established. This skin organ model has been characterized and evaluated for its suitability for radiobiological investigations. For that purpose, expression of {beta}{sub 1}-integrin following irradiation was compared in the skin organ model and in HaCaT monolayer cells (FACScan and immunohistochemistry). Furthermore, the influence of ionizing radiation on DNA fragmentation was investigated in the skin organ model (TUNEL assay). Results: the novel skin organ model showed characteristics of human skin as demonstrated by cytokeratin and Ki-67 immunoreactivity and by electron microscopy. A single dose of 5 Gy X-irradiation induced an upregulation of {beta}{sub 1}-integrin expression both in the skin organ model and in HaCaT cells. Following irradiation, {beta}{sub 1}-integrin immunoreactivity was intensified in the upper layers of the epidermis equivalent whereas it was almost absent in the deeper layers. Additionally, irradiation of the skin organ model also caused a marked increase of DNA fragmentation. Conclusion: these results demonstrate that the novel skin organ model is suitable to investigate cellular radiation effects under three-dimensional conditions. This allows to investigate radiation effects which cannot be demonstrated in monolayer cell cultures. (orig.)

Meineke, V.; Mueller, K.; Ridi, R.; Cordes, N.; Beuningen, D. van [Inst. of Radiobiology of the German Armed Forces, Munich (Germany); Koehn, F.M.; Ring, J. [Clinic of Dermatology and Allergology at Biederstein, Technical Univ. of Munich (Germany); Mayerhofer, A. [Anatomic Inst., Univ. of Munich (Germany)

2004-02-01

343

Development and evaluation of a skin organ model for the analysis of radiation effects  

International Nuclear Information System (INIS)

Background and purpose: the reaction of tissues to ionizing radiation involves alterations in cell-cell and cell-matrix interactions mediated by cellular adhesion molecules. The aim of this study was to develop and evaluate an artificial skin organ model for the analysis of radiation effects. Material and methods: a human co-culture system consisting of the spontaneously immortalized keratinocyte cell line HaCaT and primary HDFa fibroblasts embedded into a collagen sponge was established. This skin organ model has been characterized and evaluated for its suitability for radiobiological investigations. For that purpose, expression of ?1-integrin following irradiation was compared in the skin organ model and in HaCaT monolayer cells (FACScan and immunohistochemistry). Furthermore, the influence of ionizing radiation on DNA fragmentation was investigated in the skin organ model (TUNEL assay). Results: the novel skin organ model showed characteristics of human skin as demonstrated by cytokeratin and Ki-67 immunoreactivity and by electron microscopy. A single dose of 5 Gy X-irradiation induced an upregulation of ?1-integrin expression both in the skin organ model and in HaCaT cells. Following irradiation, ?1-integrin immunoreactivity was intensified in the upper layers of the epidermis equivalent whereas it was almost absent in the deeper layers. Additionally, irradiation of the skin organ model also caused a marked increase of DNA fragmo caused a marked increase of DNA fragmentation. Conclusion: these results demonstrate that the novel skin organ model is suitable to investigate cellular radiation effects under three-dimensional conditions. This allows to investigate radiation effects which cannot be demonstrated in monolayer cell cultures. (orig.)

344

Systems-Level Modeling of Cancer-Fibroblast Interaction  

OpenAIRE

Cancer cells interact with surrounding stromal fibroblasts during tumorigenesis, but the complex molecular rules that govern these interactions remain poorly understood thus hindering the development of therapeutic strategies to target cancer stroma. We have taken a mathematical approach to begin defining these rules by performing the first large-scale quantitative analysis of fibroblast effects on cancer cell proliferation across more than four hundred heterotypic cell line pairings. Systems...

Finley, S. Aidan; Bergquist, Henry; Upadhyay, Rabi; Finn, Stephen; Wadlow, Raymond C.; Wittner, Ben; Loda, Massimo; Mahmood, Umar; Ramaswamy, Sridhar

2009-01-01

345

Inhibition of protein geranylgeranylation induces apoptosis in synovial fibroblasts  

OpenAIRE

Statins, competitive inhibitors of hydroxymethylglutaryl-CoA reductase, have recently been shown to have a therapeutic effect in rheumatoid arthritis (RA). In RA, synovial fibroblasts in the synovial lining, are believed to be particularly important in the pathogenesis of disease because they recruit leukocytes into the synovium and secrete angiogenesis-promoting molecules and proteases that degrade extracellular matrix. In this study, we show a marked reduction in RA synovial fibroblast surv...

Connor, Alison M.; Berger, Stuart; Narendran, Aru; Keystone, Edward C.

2006-01-01

346

Skin Pigment  

Science.gov (United States)

... Resources for Help and Information The One-Page Merck Manual of Health Medical Terms Conversion Tables Manuals ... Disorders Overview of Skin Pigment Albinism Vitiligo Melasma Merck Manual > Patients & Caregivers > Skin Disorders > Pigment Disorders 4 ...

347

Skin diseases following a Christmas tree pattern.  

Science.gov (United States)

Pattern analysis of skin lesions is an art and a key competence of every dermatologist. Three major line patterns cover the human body-the dermatomes or Head zones, the nevoid lines of Blaschko, and the relaxed skin tension lines, or Langer lines. Head zones represent skin areas innervated from the same sensory neuronal segment or spinal nerve zone. Blaschko lines are borderlines of epidermal aberration caused by genetic mosaicism occurring in the early stages of embryogenesis. Langer lines show the direction of the lowest naturally occurring skin tension, and its thoracodorsal manifestation is the Christmas tree pattern. Here we review clinical aspects of pityriasis rosea, mycosis fungoides, stage 2 syphilis, exanthematic Kaposi sarcoma, exanthematic psoriasis, Leser-Trelat syndrome, and other primary skin diseases with a Christmas tree pattern. Secondary skin diseases, such as herpes zoster or indeterminate cell histiocytosis, may follow this pattern if they are linked to a primary skin disease by the Wolf isotopic response. PMID:21396559

Wollenberg, Andreas; Eames, Tatiana

2011-01-01

348

Skin Graft  

OpenAIRE

Skin graft is one of the most indispensable techniques in plastic surgery and dermatology. Skin grafts are used in a variety of clinical situations, such as traumatic wounds, defects after oncologic resection, burn reconstruction, scar contracture release, congenital skin deficiencies, hair restoration, vitiligo, and nipple-areola reconstruction. Skin grafts are generally avoided in the management of more complex wounds. Conditions with deep spaces and exposed bones normally require the use o...

Ruka Shimizu; Kazuo Kishi

2012-01-01

349

Skin Aging  

Science.gov (United States)

Your skin changes as you age. You might notice wrinkles, age spots and dryness. Your skin also becomes thinner and loses fat, making it ... heal, too. Sunlight is a major cause of skin aging. You can protect yourself by staying out ...

350

Accumulation of sulfate-containing acid mucopolysaccharides in I-cell fibroblasts.  

Science.gov (United States)

A rapid and sensitive papper electrophoretic assay for 35SO4-containing compounds was developed which allowed measurement of 35S-acid mucopolysaccharides synthesized by skin fibroblasts grown in the presence of inorganic 35S-sulfate. Fibroblasts from a skin explant of a patient with I-cell disease when grown in culture accumulated abnormal amounts of 35S-acid mucopolysaccharides and other, as yet unidentified, 35S-labeled compounds. Approximately 75% of the 35S-compounds accumulated by I-cell fibroblasts were not metabolized and remained in the cells after transfer to nonlabeled medium. I-cell fibroblasts differ from fibroblasts derived from classical mucopolysaccharidoses such as Hurler's and Hunter's syndromes in the amount and types of 35S-labeled acid mucopolysaccharides accumulated. I-cell fibroblasts accumulated chondroitin 4- and 6-sulfates (16 per cent), dermatan sulfate (32 per cent), heparan sulfate (32 per cent), and other unidentified 35S-compounds. The unidentified fraction was not hydrolyzed by microbial chondroitinase or heparinase. Attempts to correct the defect in I-cell fibroblasts by growth in the presence of extracts of normal cells resulted in release of only 10 per cent of the accumulated mucopolysaccharides. Under the same conditions, Hurler and Hunter fibroblasts lost over 90 per cent of accumulated mucopolysaccharides. PMID:170349

Schmickel, R D; Distler, J J; Jourdian, G W

1975-10-01

351

Light Microscopic, Electron Microscopic, and Immunohistochemical Comparison of Bama Minipig (Sus scrofa domestica) and Human Skin  

OpenAIRE

Here we sought to evaluate the possibility of using Chinese Bama miniature pig skin as a suitable animal model for human skin. Morphologic features of the skin of Bama miniature pigs resemble those of human skin, including skin layer thickness, development of a superficial vascular system, structure of the dermal–epidermal interface, and extracellular matrix. The characteristics and densities of Langerhans cells, fibroblasts, vascular endothelial cells, and mast cells were similar between B...

Liu, Yu; Chen, Jun-ying; Shang, Hai-tao; Liu, Chang-e; Wang, Yong; Niu, Rong; Wu, Jun; Wei, Hong

2010-01-01

352

Biosynthesis of collagen by fibroblasts kept in culture  

International Nuclear Information System (INIS)

The sinthesis of collagen is studied in fibroblasts of different origins with the purpose of obtaining an appropriate system for the study of its biosynthesis and processing. The percentage of collagen synthesis vary according to the fibroblast origin. Experiences are performed with fibroblasts kept in culture from: chicken - and guinea pig embryos, carragheenin - induced granulomas in adult guinea pig and from human skin. The collagen pattern synthesized after acetic acid - or saline extractions in the presence of inhibitors is also determined. This pattern is then assayed by poliacrilamide - 5% - SDS gel electrophoresis accompanied by fluorography. The importance of the cell culture system in the elucidation of collagen biosynthesis is pointed out. (M.A.)

353

Modulation of TNF-induced macrophage polarization by synovial fibroblasts.  

Science.gov (United States)

Mesenchymal stromal cells have emerged as powerful modulators of the immune system. In this study, we explored how the human macrophage response to TNF is regulated by human synovial fibroblasts, the representative stromal cell type in the synovial lining of joints that become activated during inflammatory arthritis. We found that synovial fibroblasts strongly suppressed TNF-mediated induction of an IFN-? autocrine loop and downstream expression of IFN-stimulated genes (ISGs), including chemokines CXCL9 and CXCL10 that are characteristic of classical macrophage activation. TNF induced the production of soluble synovial fibroblast factors that suppressed the macrophage production of IFN-?, and cooperated with TNF to limit the responsiveness of macrophages to IFN-? by suppressing activation of Jak-STAT signaling. Genome-wide transcriptome analysis showed that cocultured synovial fibroblasts modulate the expression of approximately one third of TNF-regulated genes in macrophages, including genes in pathways important for macrophage survival and polarization toward an alternatively activated phenotype. Pathway analysis revealed that gene expression programs regulated by synovial fibroblasts in our coculture system were also regulated in rheumatoid arthritis synovial macrophages, suggesting that these fibroblast-mediated changes may contribute to rheumatoid arthritis pathogenesis. This work furthers our understanding of the interplay between innate immune and stromal cells during an inflammatory response, one that is particularly relevant to inflammatory arthritis. Our findings also identify modulation of macrophage phenotype as a new function for synovial fibroblasts that may prove to be a contributing factor in arthritis pathogenesis. PMID:25057003

Donlin, Laura T; Jayatilleke, Arundathi; Giannopoulou, Eugenia G; Kalliolias, George D; Ivashkiv, Lionel B

2014-09-01

354

UV-induced DNA excision repair in rat fibroblasts during immortalization and terminal differentiation in vitro  

International Nuclear Information System (INIS)

UV-induced DNA excision repair was studied as DNA repair synthesis and dimer removal in rat fibroblast cultures, initiated from either dense or sparse inocula of primary cells grown from skin biopsies. During passaging in vitro an initial increase in DNA repair synthesis, determined both autoradiographically as unscheduled DNA synthesis (UDS) and by means of the BrdU photolysis assay as the number and average size of repair patches, was found to be associated with a morphological shift from small spindle-shaped to large pleiomorphic cells observed over the first twenty generations. In cell populations in growth crisis, a situation exclusively associated with thin-inoculum cultures in which the population predominantly consisted of large pleiomorphic cells, UDS was found to occur at a low level. After development of secondary cultures into immortal cell lines, both repair synthesis and morphology appeared to be the same as in the original primary spindle-shaped cells. At all passages the capacity to remove UV-induced pyrimidine dimers was found to be low, as indicated by the persistence of Micrococcus luteus UV endonuclease-sensitive sites. These results are discussed in the context of terminal differentiation and immortalization of rat fibroblasts upon establishment in vitro

355

Salmonella enterica serovar typhimurium invades fibroblasts by multiple routes differing from the entry into epithelial cells.  

Science.gov (United States)

Fibroblasts are ubiquitous cells essential to tissue homeostasis. Despite their nonphagocytic nature, fibroblasts restrain replication of intracellular bacterial pathogens such as Salmonella enterica serovar Typhimurium. The extent to which the entry route of the pathogen determines this intracellular response is unknown. Here, we analyzed S. Typhimurium invasion in fibroblasts obtained from diverse origins, including primary cultures and stable nontransformed cell lines derived from normal tissues. Features distinct to the invasion of epithelial cells were found in all fibroblasts tested. In some fibroblasts, bacteria lacking the type III secretion system encoded in the Salmonella pathogenicity island 1 displayed significant invasion rates and induced the formation of lamellipodia and filopodia at the fibroblast-bacteria contact site. Other bacterial invasion traits observed in fibroblasts were the requirement of phosphatidylinositol 3-kinase, mitogen-activated protein kinase MEK1, and both actin filaments and microtubules. RNA interference studies showed that different Rho family GTPases are targeted by S. Typhimurium to enter into distinct fibroblasts. Rac1 and Cdc42 knockdown affected invasion of normal rat kidney fibroblasts, whereas none of the GTPases tested (Rac1, Cdc42, RhoA, or RhoG) was essential for invasion of immortalized human foreskin fibroblasts. Collectively, these data reveal a marked diversity in the modes used by S. Typhimurium to enter into fibroblasts. PMID:20368348

Aiastui, Ana; Pucciarelli, M Graciela; García-del Portillo, Francisco

2010-06-01

356

Immunohistochemical characterization of human ?-irradiated skin  

International Nuclear Information System (INIS)

An immunohistochemical analysis was carried out in order to characterize the phenotypic modifications induced by ?-rays in human skin and to study the expression of some growth factors and growth factor receptors. Following radiotherapy for breast carcinoma, dermal fibroblasts of mammary skin are located superficially near the dermo-epidermal junction. They exhibit either vimentin-positive/smooth muscle cells (SMC) ?-actin-negative quiescent phenotype or vimentin-positive/SMC ?-actin-positive 'reactive' myofibroblastic phenotype but no desmin intermediate filaments. Using two polyclonal antibodies against Transforming Growth Factor ? (TGF?), we observed a specific intranuclear staining in fibroblasts and epidermal cells. Epidermal Growth Factor-Receptors (EGFR) were detected as membrane-associated in all the epidermal cell layers of irradiated skin and this pattern appears strongly associated with previous irradiation. These data suggest that complex cellular interactions are involved between epidermal and dermal cells and with extracellular matrix components, mediated by various cytokines, including TGF? and EGF-like factors

357

Biological effects of cellular stretch on human dermal fibroblasts.  

Science.gov (United States)

Pathological scars are fibroproliferative skin disorders that are characterised by the accumulation of fibroblasts and collagens. It is increasingly understood that their development and progression may be related to local skin mechanics, such as stretching. The present study evaluated the morphological and functional effects of cellular stretch on normal human dermal fibroblasts and explored the mechanotransduction mechanisms that may be involved. When fibroblasts were subjected to 24 h of cyclic axial stretching (10 cycles min(-1)), they migrated faster and for a longer distance than unstretched cells. The increased migration resulted in the cells reorienting themselves perpendicular to the direction of stretching. This was associated with reduced cellular apoptosis and unchanged proliferation. Stretching did not increase collagen synthesis but did elevate collagen degradation. These biological effects appeared to be mediated by the integrin and Wnt mechanotransduction pathways, which transmitted the mechanical stimulus via cell-substrate interactions, cell-cell junctions and indirect cell-cell communications. A better understanding of such fibroblast mechanoresponses in vitro will help the development of novel interventions that can prevent, reduce or even reverse pathological scar formation and/or progression in vivo. PMID:24055333

Huang, Chenyu; Miyazaki, Kunio; Akaishi, Satoshi; Watanabe, Atsushi; Hyakusoku, Hiko; Ogawa, Rei

2013-12-01

358

Salmonella enterica Serovar Typhimurium Invades Fibroblasts by Multiple Routes Differing from the Entry into Epithelial Cells?  

OpenAIRE

Fibroblasts are ubiquitous cells essential to tissue homeostasis. Despite their nonphagocytic nature, fibroblasts restrain replication of intracellular bacterial pathogens such as Salmonella enterica serovar Typhimurium. The extent to which the entry route of the pathogen determines this intracellular response is unknown. Here, we analyzed S. Typhimurium invasion in fibroblasts obtained from diverse origins, including primary cultures and stable nontransformed cell lines derived from normal t...

Aiastui, Ana; Pucciarelli, M. Graciela; Garci?a-del Portillo, Francisco

2010-01-01

359

Gene expression profiling reveals novel TGF? targets in adult lung fibroblasts  

OpenAIRE

Abstract Background Transforming growth factor beta (TGF?), a multifunctional cytokine, plays a crucial role in the accumulation of extracellular matrix components in lung fibrosis, where lung fibroblasts are considered to play a major role. Even though the effects of TGF? on the gene expression of several proteins have been investigated in several lung fibroblast cell lines, the global pattern of response to this cytokine in adult lung fibroblasts is still unknown. ...

Pearson Jeremy D; Leask Andrew; Denton Christopher P; Nicholson Andrew G; Veeraraghavan Srihari; Wells Athol U; Bou-Gharios George; Shi-Wen Xu; Sestini Piersante; Howat Sarah; Abraham David J; Renzoni Elisabetta A; Black Carol M; Welsh Kenneth I; du Bois Roland M

2004-01-01

360

Transformation of human fibroblasts and keratinocytes with human papillomavirus type 16 DNA.  

OpenAIRE

Human keratinocytes and fibroblasts isolated from foreskin were transformed by transfection with recombinant human papillomavirus type 16 (HPV16) DNA. The transformed cells exhibited an extended (fibroblasts) or indefinite (keratinocytes) life-span compared with that of normal controls. In addition, HS27, a human fibroblast cell line previously transfected with origin-defective simian virus 40, was successfully transfected. HPV16 sequences were stably maintained in the cells, and extensive am...

Pirisi, L.; Yasumoto, S.; Feller, M.; Doniger, J.; Dipaolo, J. A.

1987-01-01

361

Radioprotection by glutathione ester: transport of glutathione ester into human lymphoid cells and fibroblasts.  

OpenAIRE

Glutathione is not effectively transported into human lymphoid cells, normal human skin fibroblasts, and fibroblasts from patients with genetic deficiencies of gamma-glutamylcysteine synthetase or glutathione synthetase. On the other hand, the monoethyl ester of glutathione, in which the carboxyl group of the glycine residue is esterified, is readily transported into these cells and is hydrolyzed intracellularly. This leads to greatly increased cellular levels of glutathione, which often exce...

Wellner, V. P.; Anderson, M. E.; Puri, R. N.; Jensen, G. L.; Meister, A.

1984-01-01

362

The Biological Behaviors of Rat Dermal Fibroblasts Can Be Inhibited by High Levels of MMP9  

OpenAIRE

Aims. To explore the effects of the high expression of MMP9 on biological behaviors of fibroblasts. Methods. High glucose and hyperhomocysteine were used to induce MMP9 expression in skin fibroblasts. Cell proliferation was detected by flow cytometry and cell viability by CCK-8. ELISA assay was used to detect collagen (hydroxyproline) secretion. Scratch test was employed to evaluate horizontal migration of cells and transwell method to evaluate vertical migration of cells. Results. The mRNA a...

Li Yan; Diao-Zhu Lin; Chuan Yang; Sheng-Neng Xue; Juan Lei

2012-01-01

363

The matrix of human breast tumor cells is mitogenic for fibroblasts.  

OpenAIRE

The basis of the scirrhous reaction to human breast carcinoma was investigated. When normal human skin fibroblasts were plated on the preformed extracellular matrix of human breast tumor cells, a remarkable series of changes was observed. The matrix of the tumor cells was mitogenic for the fibroblasts. An increased growth rate and a fourfold increase in cell density was observed. There was also a change in cell morphology and in the pattern in which the cells grew, with an apparent loss of co...

Kao, R. T.; Hall, J.; Engel, L.; Stern, R.

1984-01-01

364

Chemotactic responses of fibroblasts to tropoelastin and elastin-derived peptides.  

OpenAIRE

Fibroblasts are known to have chemotactic responses to two components of the extracellular matrix, collagen and fibronectin. To extend these observations to other extracellular connective tissue macromolecules and their proteolytic fragments, fibroblasts from adult human skin and from late-gestation (270 d), fetal bovine ligaments were studied for chemotactic responsiveness to tropoelastin and elastin-derived peptides. Bovine ligament tropoelastin and elastin-derived peptides, generated from ...

Senior, R. M.; Griffin, G. L.; Mecham, R. P.

1982-01-01

365

Co?culture with human fetal epidermal keratinocytes promotes proliferation and migration of human fetal and adult dermal fibroblasts.  

Science.gov (United States)

The repair strategy for the healing of skin wounds in fetuses differs from that in adults. Proliferation and migration of dermal fibroblasts are the main mechanisms associated with skin wound healing, as well as the complex interactions between epidermal keratinocytes (KCs) and dermal fibroblasts. In order to investigate the effects of fetal skin epidermal KCs on fetal and adult human dermal fibroblasts, KCs and fibroblasts were isolated from the skin tissue of mid?gestational human fetuses and adults, and co?cultured using a Transwell® system. When fetal mid?gestational KCs were co?cultured with either fetal or adult dermal fibroblasts, the proliferative and migratory potential of the fibroblasts was significantly enhanced. Furthermore, these phenotypic changes were concomitant with the upregulation of numerous proteins including mouse double minute 2 homolog, cyclin B1, phospho?cyclin?dependent kinase 1, phospho?extracellular signal?regulated kinase, and phospho?AKT, along with C?X?C chemokine receptor 4, phospho?p38 mitogen activated protein kinase, matrix metalloproteinase (MMP)?2 and MMP?9. Notably, no significant differences were observed between fetal and adult dermal fibroblasts in their responses to fetal mid?gestational epidermal KCs, indicating that the cells from these two developmental stages respond in a similar manner to co?culture with KCs. PMID:25351528

Wang, Zhe; Liu, Xiaoyu; Zhang, Dianbao; Wang, Xiliang; Zhao, Feng; Shi, Ping; Pang, Xining

2015-02-01

366

Morfometria de fibroblastos e fibrócitos durante o processo cicatricial na pele de coelhos da raça Nova Zelândia Branco tratados com calêndula / Morphometry of fibroblasts and fibrocytes during wound healing in the skin of rabbits of the New Zeland White breed treated with marigold  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese O objetivo deste estudo foi avaliar a capacidade cicatrizante da calêndula (Calendula officinalis L.) sobre feridas cutâneas experimentais, em 15 coelhos, distribuídos em três grupos denominados: excipiente, calêndula e controle. Cada animal foi submetido à uma incisão cirúrgica de 6cm de compriment [...] o, lateral à coluna vertebral e suturada no padrão U. Os produtos avaliados foram colocados sobre as incisões durante sete dias na quantidade de 0,1ml (loção cremosa não-iônica - grupo excipiente; tintura de calêndula a 5% - grupo calêndula) e nos animais do grupo controle não se utilizou nenhum produto. A biópsia de pele foi realizada no 1°, 3°, 5° e 7° dia após a incisão cirúrgica para avaliação morfométrica do processo cicatricial, analisando-se o número de fibroblastos e fibrócitos. A morfometria foi realizada por meio de microscópio óptico adaptado a um sistema computadorizado de análise de imagens. De acordo com os resultados, a calêndula propiciou obtenção dos maiores valores médios das células envolvidas no processo cicatricial, os fibroblastos, deduzindo que a mesma, inferiu uma resposta mais satisfatória na cicatrização em relação aos demais tratamentos. Abstract in english The aim of this study was to evaluate the scarring capability of marigold (Calendula officinalis L.) on experimental skin wounds in 15 rabbits, distributed in three groups: excipient, marigold and control. Each animal was subjected to a surgical incision measuring 6cm in length, laterally to the spi [...] nal column and sutured in U-shape. Products evaluated were placed on the incisions for 7 days, at a rate of 0.1ml (nonionic creamy lotion - excipient group; 5% marigold extract) and no treatment was provided to control animals. Skin biopsy was performed on 1, 3, 5, and 7 days after wounding, for morphometric and cicatricial process evaluations. The morphometry was performed with an optical microscope adapted to a computadorized picture analysis system. The results showed that marigold allowed the highest growth rate in cells directly involved in the cicatricial process, the fibroblasts and fibrocytes and can therefore be considered the most satisfactory on the wound healing in comparison to the other treatments.

Leonardo de Oliveira, Pagnano; Silvana Martinez, Baraldi-Artoni; Maria Rita, Pacheco; Edanir dos, Santos; Daniela, Oliveira; Jeffrey Frederico, Lui.

1662-16-01

367

Morfometria de fibroblastos e fibrócitos durante o processo cicatricial na pele de coelhos da raça Nova Zelândia Branco tratados com calêndula Morphometry of fibroblasts and fibrocytes during wound healing in the skin of rabbits of the New Zeland White breed treated with marigold  

Directory of Open Access Journals (Sweden)

Full Text Available O objetivo deste estudo foi avaliar a capacidade cicatrizante da calêndula (Calendula officinalis L. sobre feridas cutâneas experimentais, em 15 coelhos, distribuídos em três grupos denominados: excipiente, calêndula e controle. Cada animal foi submetido à uma incisão cirúrgica de 6cm de comprimento, lateral à coluna vertebral e suturada no padrão U. Os produtos avaliados foram colocados sobre as incisões durante sete dias na quantidade de 0,1ml (loção cremosa não-iônica - grupo excipiente; tintura de calêndula a 5% - grupo calêndula e nos animais do grupo controle não se utilizou nenhum produto. A biópsia de pele foi realizada no 1°, 3°, 5° e 7° dia após a incisão cirúrgica para avaliação morfométrica do processo cicatricial, analisando-se o número de fibroblastos e fibrócitos. A morfometria foi realizada por meio de microscópio óptico adaptado a um sistema computadorizado de análise de imagens. De acordo com os resultados, a calêndula propiciou obtenção dos maiores valores médios das células envolvidas no processo cicatricial, os fibroblastos, deduzindo que a mesma, inferiu uma resposta mais satisfatória na cicatrização em relação aos demais tratamentos.The aim of this study was to evaluate the scarring capability of marigold (Calendula officinalis L. on experimental skin wounds in 15 rabbits, distributed in three groups: excipient, marigold and control. Each animal was subjected to a surgical incision measuring 6cm in length, laterally to the spinal column and sutured in U-shape. Products evaluated were placed on the incisions for 7 days, at a rate of 0.1ml (nonionic creamy lotion - excipient group; 5% marigold extract and no treatment was provided to control animals. Skin biopsy was performed on 1, 3, 5, and 7 days after wounding, for morphometric and cicatricial process evaluations. The morphometry was performed with an optical microscope adapted to a computadorized picture analysis system. The results showed that marigold allowed the highest growth rate in cells directly involved in the cicatricial process, the fibroblasts and fibrocytes and can therefore be considered the most satisfactory on the wound healing in comparison to the other treatments.

Leonardo de Oliveira Pagnano

2008-09-01

368

Cadherin-11 regulates fibroblast inflammation  

OpenAIRE

Fibroblasts are important participants in inflammation. Although not leukocytes, their capacity to produce cytokines, chemokines, and other inflammatory factors locally in tissues suggests that they can contribute to inflammatory diseases. For example, fibroblasts in a rheumatoid arthritis (RA) joint are a dominant source of IL-6 and RANKL in the synovium, both of which are therapeutic targets for inflammation and bone erosion. Previously, we found that fibroblasts can be targeted by mAb dire...

Chang, Sook Kyung; Noss, Erika H.; Chen, Mei; Gu, Zhizhan; Townsend, Kirk; Grenha, Rosa; Leon, Luis; Lee, Soo Young; Lee, David M.; Brenner, Michael B.

2011-01-01

369

Biology of Fibroblasts and Myofibroblasts  

OpenAIRE

Despite their importance in fibrosis, the origin of fibroblasts and the genesis of the various subpopulations characterized by distinct phenotypes remain unclear. Various studies have described distinct and relatively stable phenotypes in fibroblasts isolated from lung tissue undergoing remodeling, which were not present in the normal intact tissue. This indicates a process by which these distinct fibroblast subpopulations could arise de novo from resident lung progenitors or precursor cells ...

Phan, Sem H.

2008-01-01

370

Skin optics  

Energy Technology Data Exchange (ETDEWEB)

Quantitative dosimetry in the treatment of skin disorders with (laser) light requires information on propagation of light in the skin related to the optical properties of the individual skin layers. This involves the solution of the integro-differential equation of radiative transfer in a model representing skin geometry, as well as experimental methods to determine the optical properties of each skin layer. These activities are unified under the name skin optics. This paper first reviews the current status of tissue optics, distinguishing between the cases of: dominant absorption, dominant scattering, and scattering about equal to absorption. Then, previously published data as well as some current unpublished data on (human) stratum corneum, epidermis and dermis, have been collected and/or (re)analyzed in terms of absorption coefficient, scattering coefficient, and anisotropy factor of scattering. The results are that the individual skin layers show strongly forward scattering (anisotropy factors between 0.7 and 0.9). The absorption and scattering data show that for all wavelengths considered scattering is much more important than absorption. Under such circumstances, solutions to the transport equation for a multilayer skin model and finite beam laser irradiation are currently not yet available. Hence, any quantitative dosimetry for skin treated with (laser) light is currently lacking.

van Gemert, M.J.; Jacques, S.L.; Sterenborg, H.J.; Star, W.M.

1989-12-01

371

Increased fibroblast functionality on CNN2-loaded titania nanotubes  

Directory of Open Access Journals (Sweden)

Full Text Available Hongbo Wei*, Shuyi Wu*, Zhihong Feng, Wei Zhou, Yan Dong, Guofeng Wu, Shizhu Bai, Yimin Zhao Department of Prosthodontics, School of Stomatology, Fourth Military Medical University, Xi'an, People's Republic of China *These authors contributed equally to this workAbstract: Infection and epithelial downgrowth are major problems associated with maxillofacial percutaneous implants. These complications are mainly due to the improper closure of the implant–skin interface. Therefore, designing a percutaneous implant that better promotes the formation of a stable soft tissue biologic seal around percutaneous sites is highly desirable. Additionally, the fibroblast has been proven to play an important role in the formation of biologic seals. In this study, titania nanotubes were filled with 11.2 kDa C-terminal CCN2 (connective tissue growth factor fragment, which could exert full CCN2 activity to increase the biological functionality of fibroblasts. This drug delivery system was fabricated on a titanium implant surface. CCN2 was loaded into anodized titania nanotubes using a simplified lyophilization method and the loading efficiency was approximately 80%. Then, the release kinetics of CCN2 from these nanotubes was investigated. Furthermore, the influence of CCN2-loaded titania nanotubes on fibroblast functionality was examined. The results revealed increased fibroblast adhesion at 0.25, 0.5, 1, 2, 4, and 24 hours, increased fibroblast viability over the course of 5 days, as well as enhanced actin cytoskeleton organization on CCN2-loaded titania nanotubes surfaces compared to uncoated, unmodified counterparts. Therefore, the results from this in vitro study demonstrate that CCN2-loaded titania nanotubes have the ability to increase fibroblast functionality and should be further studied as a method of promoting the formation of a stable soft tissue biologic seal around percutaneous sites.Keywords: anodization, titania nanotubes, adhesion, connective tissue growth factor, fibroblast

Wei HB

2012-02-01

372

Punch-wounded, fibroblast populated collagen matrices: a novel approach for studying cytoskeletal changes in three dimensions by confocal laser scanning microscopy.  

Science.gov (United States)

Depending on growth conditions and cell type, collagen matrices populated with viable cells, i.e. commonly with fibroblasts, contract in a manner resembling wound contraction in vivo. If matrix cultures can be grown to provide the environment of contracting wounds in vitro, other conditions may be established under which fibroblasts grow reminiscent of those in normal dermis. Wounding such dermal equivalents may then initiate cells to change their phenotype in space and time in an in vivo-like environment. In turn, such a system should allow to study the underlying cytoskeletal changes at the onset of tissue repair and beyond. To test this hypothesis, we established the conditions for human skin-derived fibroblasts (KD cells) to grow within collagen matrices without contraction. We then excised from the center of such "attached, low-contracting dermal equivalents" (ALDE) "punch biopsies" with a diameter of 1 mm, and monitored the cell's shape and their microtubular networks and F-actin-containing structures over time by i) conventional fluorescence microscopy and ii) by confocal laser scanning microscopy in combination with optical sectioning and volume rendering software. Prior to wounding and in non-wounded controls up to 8 days post seeding (ps), cells predominantly exhibited an elongated, spindle-shaped morphology with distinct microtubular networks and F-actin-containing structures. Wounding induced most of the fibroblasts lining the wound edge to immediately round up. The round cells still revealed a microtubular network but only diffuse labeling for F-actin. One day post wounding (pw), these fibroblasts had resumed their spindle-shaped structure. They displayed the microtubular network and again thin F-actin-containing structures. From 2 days pw on and up to 6 days pw, the number of fibroblasts in the wound zone had increased, forming dense, multilayered patches oriented parallel to the wound surface, and the cells lining the wound margin showed the most extensive and massive F-actin-containing stress fiber-like structures. Quantification of the cell densities at the wound margin and in adjacent zones corroborated this increase, which is reminiscent of the fibroblasts migrating to the wound edge in the early phase of connective tissue repair. PMID:9084981

Baschong, W; Sütterlin, R; Aebi, U

1997-03-01

373

Heparin fragments modulate the collagen phenotype of fibroblasts from radiation-induced subcutaneous fibrosis  

International Nuclear Information System (INIS)

Acute local gamma irradiation of porcine skin induces, as in human skin, an extensive and mutilating sclerosis characterized by continuous expansion of the fibrosis invading the adjacent muscle and by accumulation of the macromolecular components of the extracellular matrix. Collagen synthesis, content, and types were studied in the presence of heparin fragments (100 micrograms/10(6) cells) in the culture medium, by measuring the incorporation of the radiolabeled precursor [3H]proline into confluent primary cultures of porcine fibroblasts obtained from normal and irradiated fibrotic dermis. Enhancement in collagen biosynthesis and deposition and preferential increase in collagen type III synthesis were observed in fibrotic fibroblast cultures when compared to those in normal dermis fibroblasts. The total collagen synthesis and the rate of collagen hydroxylation appear unmodified by heparin fragments both in normal and in fibrotic fibroblast cultures. But heparin fragments induce a 10- and 2-fold decrease, respectively, in collagen type III and type V syntheses by fibrosis fibroblasts. As only minor effects upon collagen type III and V are observed in cultures of normal dermis fibroblasts, these results highly suggest that heparin fragments are capable of specifically modulating the collagen phenotype of fibroblasts derived from radiation-induced dermis fibrosis and thus are able to regulate the fibrotic process

374

Dynamic Tensile Properties of Human Skin  

OpenAIRE

The mechanical properties of skin are important for a number of applications including surgery, dermatology, impact biomechanics and forensic science. Studies have shown that the anisotropic effects of skin have been linked to sample orientation with respect to contour lines of tension, i.e. the Langer’s lines. There have been numerous studies undertaken to calculate the influence of Langer’s lines on the mechanical properties of human skin at quasistatic strain rates; however, it is rela...

Gallagher, A. J.; Ni? Annaidh, Aisling; Bruye?re, Karine; et al.

2012-01-01

375

Aging Skin  

Science.gov (United States)

... spend a lot of money each year on skin care products that promise to erase wrinkles, lighten age spots, and eliminate itching, flaking, or redness. But the simplest and cheapest way to keep your skin healthy and young looking is to stay out ...

376

Denatured Collagen Modulates the Phenotype of Normal and Wounded Human Skin Equivalents  

OpenAIRE

Epithelial–mesenchymal interactions are known to play an important role in modulating homeostasis and repair. However, it remains unclear how the composition of the extracellular matrix may regulate the ability of dermal fibroblasts to engage in such cross talk. To address this, we studied how fibroblast phenotype was linked to the behavior of normal and wounded human skin equivalents (HSE) by comparing human dermal fibroblasts (HDF) incorporated into the three-dimensional tissues to those ...

Egles, Christophe; Shamis, Yulia; Mauney, Joshua R.; Volloch, Vladimir; Kaplan, David L.; Garlick, Jonathan A.

2008-01-01

377

Accelerated telomere shortening and replicative senescence in human fibroblasts overexpressing mutant and wild-type lamin A  

International Nuclear Information System (INIS)

LMNA mutations are responsible for a variety of genetic disorders, including muscular dystrophy, lipodystrophy, and certain progeroid syndromes, notably Hutchinson-Gilford Progeria. Although a number of clinical features of these disorders are suggestive of accelerated aging, it is not known whether cells derived from these patients exhibit cellular phenotypes associated with accelerated aging. We examined a series of isogenic skin fibroblast lines transfected with LMNA constructs bearing known pathogenic point mutations or deletion mutations found in progeroid syndromes. Fibroblasts overexpressing mutant lamin A exhibited accelerated rates of loss of telomeres and shortened replicative lifespans, in addition to abnormal nuclear morphology. To our surprise, these abnormalities were also observed in lines overexpressing wild-type lamin A. Copy number variants are common in human populations; those involving LMNA, whether arising meiotically or mitotically, might lead to progeroid phenotypes. In an initial pilot study of 23 progeroid cases without detectable WRN or LMNA mutations, however, no cases of altered LMNA copy number were detected. Nevertheless, our findings raise a hypothesis that changes in lamina organization may cause accelerated telomere attrition, with different kinetics for overexpession of wild-type and mutant lamin A, which leads to rapid replicative senescence and progroid phenotypes

378

???????????????? Research on Detecting Skin Grooves in Skin Aging Analysis  

Directory of Open Access Journals (Sweden)

Full Text Available ???????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????Detecting skin grooves plays an important role in the objective quantification of human skin aging, and the detection accuracy directly influences the subsequent computation of two-dimensional geometrical characteristics of the skin surface. Based on the combined control strategies consisting of data-driven and model-driven control, a novel seg-mentation approach to skin grooves detection was proposed in this paper. In data-driven control, the rough segmentation result was obtained by applying watershed transform to the enhanced image. In model-driven control, the information acquired from the data-driven control and prior knowledge derived from sense of vision were used for constructing a region merging model. The model was used for removing redundant watershed lines. Subjective and objective evalua-tions demonstrate the favorable performance of the proposed method in detecting precisely skin grooves and restraining false skin grooves.

???

2012-03-01

379

The effect of pantothenic acid deficiency on keratinocyte proliferation and the synthesis of keratinocyte growth factor and collagen in fibroblasts.  

Science.gov (United States)

It has been reported that pantothenic acid (vitamin B5) and panthenol, an alcohol derivative of pantothenic acid, have beneficial moisturizing effects on the skin. However, few studies have investigated the mechanism of action of pantothenic acid on skin tissues. We tried to clarify the role of pantothenic acid on skin function by using keratinocytes and fibroblasts. The depletion of pantothenic acid from the culture medium suppressed keratinocyte proliferation and promoted differentiation. Moreover, pantothenic acid depletion decreased the synthesis of keratinocyte growth factor and procollagen 4a2 in fibroblasts. These results suggest that pantothenic acid is essential for maintaining keratinocyte proliferation and differentiation. PMID:21258175

Kobayashi, Daisaku; Kusama, Miho; Onda, Masaaki; Nakahata, Norimichi

2011-01-01

380

Keratinocyters påverkan på fibroblasters aktivitet  

OpenAIRE

In the healing of cutaneous wounds, paracrine communication between keratinocytes and fibroblasts regulates cell differentiation, proliferation and synthesis of extracellular matrix. Deficient epidermal coverage, as seen in burn-wounds, frequently results in hypertrophic scars. Previous studies suggest that keratinocytes downregulate the production of collagen and profibrotic factors in fibroblasts. We hypothesized that keratinocytes downregulate the expression of the profibrotic factor conne...

Nowinski, Daniel

2005-01-01

381

Effects of lunar and mars dust simulants on HaCaT keratinocytes and CHO-K1 fibroblasts  

Science.gov (United States)

Exposure to lunar dust during Apollo missions resulted in occasional reports of ocular, respiratory and dermal irritations which showed that lunar dust has a risk potential for human health. This is caused by its high reactivity as well as its small size, leading to a wide distribution also inside habitats. Hence, detailed information regarding effects of extraterrestrial lunar dusts on human health is required to best support future missions to moon, mars or other destinations. In this study, we used several methods to assess the specific effects of extraterrestrial dusts onto mammalian skin by exposing HaCaT keratinocytes and CHO-K1 fibroblasts to dusts simulating lunar or mars soils. These particular cell types were chosen because the skin protects the human body from potentially harmful substances and because a well orchestrated program ensures proper wound healing. Keratinocytes and fibroblasts were exposed to the dusts for different durations of time and their effects on morphology and viability of the cells were determined. Cytotoxicity was measured using the MTT assay and by monitoring culture impedance, while phalloidin staining of the actin cytoskeleton was performed to address structural integrity of the cells which was also investigated by propidium iodide intake. It was found that the effects of the two types of dust simulants on the different features of both cell lines varied to a considerable extent. Moreover, proliferation of HaCaT keratinocytes, as analyzed by Ki67 labeling, was suppressed in sub-confluent cultures exposed to lunar dust simulant. Furthermore, experimental evidence is provided for a delay in regeneration of keratinocyte monolayers from scratch-wounding when exposed to lunar dust simulant. The obtained results will facilitate further investigations of dust exposure during wound healing and will ease risk assessment studies e.g., for lunar lander approaches. The investigations will help to determine safety measures to be taken during extraterrestrial expeditions in order to minimize risks to human health associated with exposure of human skin to dust contaminants.

Rehders, Maren; Grosshäuser, Bianka B.; Smarandache, Anita; Sadhukhan, Annapurna; Mirastschijski, Ursula; Kempf, Jürgen; Dünne, Matthias; Slenzka, Klaus; Brix, Klaudia

2011-04-01

382

Skin and Sports  

Science.gov (United States)

In this lesson, students learn about the importance of proper protection from common skin conditions when they engage in sports-related activities. This lesson draws attention to fact that the body's own first line of defense against infectious agents is to keep them from entering or settling in the body. The students break into groups to provide a list of risk factors for each sports-related activity. They come together and compare notes. This sparks the lesson and instruction on how one should protect the skin when participating in sports. Links to other resources for further inquiry are given.

American Association for the Advancement of Science (;)

2006-02-13

383

HSP27 as a biomarker for predicting skin irritation in human skin and reconstructed organotypic skin model.  

Science.gov (United States)

In vitro alternative tests aiming at replacing the traditional animal test for predicting the irritant potential of chemicals have been developed, but the assessing parameters or endpoints are still not sufficient. To discover novel endpoints for skin irritation responses, 2DE-based proteomics was used to analyze the protein expression in human skin exposed to sodium lauryl sulfate (SLS) following the test protocol of the European Centre for the Validation of Alternative Methods (ECVAM) in the present study. HSP27 was up-regulated most significantly among the eight identified proteins, consistent with our previous reports. Acid and basic chemicals were applied on human skin for further validation and results showed that the up-regulated expression of HSP27 was induced in 24h after the exposure. Skin-equivalent constructed with fibroblasts, basement membrane and keratinocytes was used to investigate the potential of HSP27 as a biomarker or additional endpoint for the hazard assessment of skin irritation. Our skin-equivalent (Reconstructed Organotypic Skin Model, ROSM) had excellent epidermal differentiation and was suitable for the skin irritation test. HSP27 also displayed an up-regulated expression in the ROSM in 24h after the irritants exposure for 15min. All these results suggest that HSP27 may represent a potential marker or additional endpoint for the hazard assessment of skin irritation caused by chemical products. PMID:24503015

Chen, Hongxia; Li, Shuhua; Meng, Tian; Zhang, Lei; Dai, Taoli; Xiang, Qi; Su, Zhijian; Zhang, Qihao; Huang, Yadong

2014-04-21

384

Development of nanofibrous scaffolds containing gum tragacanth/poly (?-caprolactone) for application as skin scaffolds.  

Science.gov (United States)

Outstanding wound healing activity of gum tragacanth (GT) and higher mechanical strength of poly (?-caprolactone) (PCL) may produce an excellent nanofibrous patch for either skin tissue engineering or wound dressing application. PCL/GT scaffold containing different concentrations of PCL with different blend ratios of GT/PCL was produced using 90% acetic acid as solvent. The results demonstrated that the PCL/GT (3:1.5) with PCL concentration of 20% (w/v) produced nanofibers with proper morphology. Scanning electron microscopy (SEM) and differential scanning calorimetry (DSC) were utilized to characterize the nanofibers. Surface wettability, functional groups analysis, porosity and tensile properties of nanofibers were evaluated. Morphological characterization showed that the addition of GT to PCL solution results in decreasing the average diameter of the PCL/GT nanofibers. However, the hydrophilicity increased in the PCL/GT nanofibers. Slight increase in melting peaks was observed due to the blending of PCL with GT nanofibers. PCL/GT nanofibers were used for in vitro cell culture of human fibroblast cell lines AGO and NIH 3T3 fibroblast cells. MTT assay and SEM results showed that the biocomposite PCL/GT mats enhanced the fibroblast adhesion and proliferation compared to PCL scaffolds. The antibacterial activity of PCL/GT and GT nanofibers against Staphylococcus aureus and Pseudomonas aeruginosa was also examined. PMID:25579898

Ranjbar-Mohammadi, Marziyeh; Bahrami, S Hajir

2015-03-01

385

Gene targeting in adult rhesus macaque fibroblasts  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Gene targeting in nonhuman primates has the potential to produce critical animal models for translational studies related to human diseases. Successful gene targeting in fibroblasts followed by somatic cell nuclear transfer (SCNT has been achieved in several species of large mammals but not yet in primates. Our goal was to establish the protocols necessary to achieve gene targeting in primary culture of adult rhesus macaque fibroblasts as a first step in creating nonhuman primate models of genetic disease using nuclear transfer technology. Results A primary culture of adult male fibroblasts was transfected with hTERT to overcome senescence and allow long term in vitro manipulations. Successful gene targeting of the HPRT locus in rhesus macaques was achieved by electroporating S-phase synchronized cells with a construct containing a SV40 enhancer. Conclusion The cell lines reported here could be used for the production of null mutant rhesus macaque models of human genetic disease using SCNT technology. In addition, given the close evolutionary relationship and biological similarity between rhesus macaques and humans, the protocols described here may prove useful in the genetic engineering of human somatic cells.

Wolf Don P

2008-03-01

386

Spatiotemporal expression of periostin during skin development and incisional wound healing: lessons for human fibrotic scar formation  

OpenAIRE

Differentiation of fibroblasts to myofibroblasts and collagen fibrillogenesis are two processes essential for normal cutaneous development and repair, but their misregulation also underlies skin-associated fibrosis. Periostin is a matricellular protein normally expressed in adult skin, but its role in skin organogenesis, incisional wound healing and skin pathology has yet to be investigated in any depth. Using C57/BL6 mouse skin as model, we first investigated periostin protein and mRNA spati...

Zhou, Hong-ming; Wang, Jian; Elliott, Christopher; Wen, Weiyan; Hamilton, Douglas W.; Conway, Simon J.

2010-01-01

387

Skin Cancer in Skin of Color  

OpenAIRE

Skin cancers in skin of color often present atypically or with advanced stage in comparison to Caucasian patients. Health care providers must maintain a high index of suspicion when examining skin lesions in skin of color.

Bradford, Porcia T.

2009-01-01

388

In vitro 3D full thickness skin equivalent tissue model using silk and collagen biomaterials  

OpenAIRE

Current approaches to develop skin equivalents often only include the epidermal and dermal components. Yet, full thickness skin includes the hypodermis, a layer below the dermis of adipose tissue containing vasculature, nerves and fibroblasts, necessary to support the epidermis and dermis. In the present study, we developed a full thickness skin equivalent including an epidermis, dermis and hypodermis that could serve as an in vitro model for studying skin development, disease or as a platfor...

Bellas, Evangelia; Seiberg, Miri; Garlick, Jonathan; Kaplan, David L.

2012-01-01

389

Divergent control of Cav-1 expression in non-cancerous Li-Fraumeni syndrome and human cancer cell lines  

OpenAIRE

Li-Fraumeni syndrome (LFS) is primarily characterized by development of tumors exhibiting germ-line mutations in the p53 gene. Cell lines developed from patients of a LFS family have decreased p53 activity as evidenced by the absence of apoptosis upon etoposide treatment. To test our hypothesis that changes in gene expression beyond p53 per se are contributing to the development of tumors, we compared gene expression in non-cancerous skin fibroblasts of LFS-affected (p53 heterozygous) vs. non...

Sherif, Zaki A.; Sultan, Ahmed S.

2013-01-01

390

Skin Tightening  

Science.gov (United States)

... AAD pamphlet) References: Alexiades-Armenakas MR, Dover JS, Arndt KA. “Laser Therapy.” In: Bolognia JL, Jorizzo JL, ... 2008. p. 2113. Alexiades-Armenakas MR, Dover JS, Arndt KA. “The spectrum of laser skin resurfacing: nonablative, ...

391

Skinning maps  

CERN Document Server

Let M be a hyperbolic 3-manifold with nonempty totally geodesic boundary.