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

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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

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Alteration of Skin Properties with Autologous Dermal Fibroblasts  

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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

3

Alteration of skin properties with autologous dermal fibroblasts.  

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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. PMID:24828202

Thangapazham, Rajesh L; Darling, Thomas N; Meyerle, Jon

2014-01-01

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Attenuated prostaglandin formation in peroxisomal-deficient human skin fibroblasts.  

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Peroxisomal-deficient skin fibroblasts from patients with Zellweger's syndrome or infantile Refsum's disease produced fewer prostaglandins than normal skin fibroblasts. Radioimmunoassay indicated a 45-55% decrease in prostaglandin E2 (PGE2) production when Zellweger's fibroblasts were incubated with arachidonic acid. This deficiency was not overcome by pretreatment of the Zellweger's fibroblasts with media containing arachidonic acid, and it was not due to channeling of arachidonic acid into ...

Gordon, J. A.; Warnock, L. J.; Spector, A. A.

1993-01-01

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Protoporphyrin-induced photodamage: studies using cultured skin fibroblasts  

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An new system is described for the study of protoporphyrin-induced photodamage of cells. This system differs from those previously described in that fibroblasts are induced to synthesize protoporphyrin from its precursor delta-aminolevulinic acids. Fibroblasts were cultured from skin biopsies of 6 normal individuals and 6 patients with protoporphyria. All cell lines in both groups accumulated protoporphyrin when incubated with delta-aminolevulinic acid in the absence of iron. Irradiation for 2 min with long-wave UV light caused death of cells which had accumulated protoporphyrin, but not of cells which had been incubated without delta-aminolevulinic acid. Cell damage could be quantitatively assessed by the release of chromium-51 into the medium. Examination of irradiated protoporphyrin-rich fibroblasts by electron microscopy revealed no significant differences between lines from patients with protoporphyria and normal individuals. The earliest indications of photodamage were rarifaction of the mitochondrial matrix with dilatation of cristae, dilatation of endoplasmic reticulum, and loss of plasma membrane integrity. Condensation of the cells correlated with cell death. These structural alterations suggest a generalized cellular injury.

Latham, P.S. (Maryland Univ., Baltimore (USA). Hospital); Bloomer, J.R. (Minnesota Univ., Minneapolis (USA))

1983-05-01

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Protoporphyrin-induced photodamage: studies using cultured skin fibroblasts  

International Nuclear Information System (INIS)

An new system is described for the study of Protoporphyrin-induced photodamage of cells. This system differs from those previously described in that fibroblasts are induced to synthesize protoporphyrin from its precursor delta-aminolevulinic acids. Fibroblasts were cultured from skin biopsies of 6 normal individuals and 6 patients with protoporphyria. All cell lines in both groups accumulated protoporphyrin when incubated with delta-aminolevulinic acid in the absence of iron. Irradiation for 2 min with long-wave UV light caused death of cells which had accumulated protoporphyrin, but not of cells which had been incubated without delta-aminolevulinic acid. Cell damage could be quantitatively assessed by the release of chromium-51 into the medium. Examination of irradiated protoporphyrin-rich fibroblasts by electron microscopy revealed no significant differences between lines from patients with protoporphyria and normal individuals. The earliest indications of photodamage were rarifaction of the mitochondrial matrix with dilatation of cristae, dilatation of endoplasmic reticulum, and loss of plasma membrane integrity. Condensation of the cells correlated with cell death. These structural alterations suggest a generalized cellular injury. (author)

1983-01-01

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Dynamics of Micronuclei in Rat Skin Fibroblasts after X Irradiation  

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In a previous study, we demonstrated DNA damage, expressed as micronuclei, in binucleate dermal fibroblasts obtained from human skin 2-9 weeks after fractionated radiotherapy. Here we assessed micronuclei in X-irradiated skin fibroblasts from 9-14-week-old female Lewis rats as a function of time after a single dose of radiation to determine the lifetime of such damage in the skin. After irradiation with 5, 10, 15 and 18 Gy, formation of micronuclei at 1 day or 2 months postirradiation increas...

Kaspler, P.; Pintilie, M.; Hill, R. P.

2009-01-01

8

Testosterone metabolism of fibroblasts grown from prostatic carcinoma, benign prostatic hyperplasia and skin fibroblasts  

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The metabolism of [1,2,6,7-3H]testosterone was assessed in fibroblast monolayers derived from tissue of 5 prostates with benign hyperplasia (BPH), 4 prostates with carcinoma (PC), and 3 biopsy samples of skin, 2 nongenital skin (NG) and 1 genital skin. The following metabolites could be identified: androstanedione androstenedione, dihydrotestosterone, androsterone, epiandrosterone, androstane-3 alpha, 17 beta-diol and androstane-3 beta, 17 beta-diol. Testosterone was metabolized much more rapidly in fibroblasts originating from prostatic tissue than in fibroblasts derived from NG. A significantly higher formation of 5 alpha-androstanes and 3 alpha-hydroxysteroids could be observed in fibroblasts from BPH as compared to PC. 17-ketosteroid formation exceeded 5 alpha-androstane formation in BPH, whereas 5 alpha-reduction was the predominant pathway in fibroblasts grown from PC and NG. Since testosterone metabolism in fibroblasts of prostatic origin therefore resembles in many aspects that in whole prostatic tissue, fibroblasts grown from prostatic tissues might be a valuable tool for further investigation of the pathogenesis of human BPH and PC

1982-01-01

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Studies of the in vivo radiosensitivity of human skin fibroblasts  

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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 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

2007-07-01

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Acetyl-11-keto-beta-boswellic acid and arsenic trioxide regulate the productions and activities of matrix metalloproteinases in human skin fibroblasts and human leukemia cell line THP-1  

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Full Text Available Objective: In order to reveal the treatment mechanism of Chinese medicine with the effect of activating blood and resolving putridity, we selected acetyl-11-keto-beta-boswellic acid (AKBA and arsenic trioxide (ATO, the main monomeric components of frankincense and arsenolite which are two most commonly used Chinese medicine with effect of activating blood and resolving putridity. We combined AKBA and ATO as a compound, and explored its regulatory role in productions and activities of matrix metalloproteinase (MMP-1, MMP-2 and MMP-9 in human skin fibroblasts (HSFbs and human acute monocytic leukemia cell line THP-1 in inflammatory state.Methods: In order to simulate the inflammatory micro-environment of chronic wounds, we established 3 cell models: HSFb model activated by tumor necrosis factor-alpha (TNF-?, THP-1 cell model activated by phorbol-12-myristate-13-acetate (PMA and HSFb-THP-1 cell coculture system. AKBA and ATO were cocultured with these cell models. Enzyme-linked immunosorbent assay (ELISA, gelatin zymography assay and reverse transcription-polymerase chain reaction (RT-PCR were used to test the secretions, activities and mRNA expressions of MMP-1, MMP-2 and MMP-9. In the study of the regulatory mechanism of AKBA and ATO on MMPs, AKBA and ATO were cocultured with the cell models. ELISA was used to test the secretions of TNF-? and interleukin-1beta (IL-? and Western blot was used to test the phosphorylation levels of extracellular signal-regulated kinases 1 and 2 (ERK1/2 and p38 mitogen-activated proteinkinase (p38MAPK. Results: Compound of AKBA and ATO inhibited MMP-1, MMP-2 and MMP-9 mRNA expressions, secretions and activities respectively in HSFbs and THP-1 cells in inflammatory state (P?0.05, P?0.01. Also compound of AKBA and ATO inhibited secretions of TNF-? and IL-1? in THP-1 cells and cell coculture system (P?0.01. It also decreased the phosphorylation of ERK1/2 and p38 MAPK in HSFbs and THP-1 cells (P?0.05, P?0.01. Conclusion: The combined use of AKBA and ATO which in line with the rule of activating blood and resolving putridity inhibits fibroblasts and inflammatory cells in producing MMPs in inflammatory state through inhibiting the release of inflammatory factors and MAPK cascade pathway.

Ya-hui Liang

2010-11-01

11

Monomethylarsonous acid inhibited endogenous cholesterol biosynthesis in human skin fibroblasts.  

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Human exposure to arsenic in drinking water is a widespread public health concern, and such exposure is known to be associated with many human diseases. The detailed molecular mechanisms about how arsenic species contribute to the adverse human health effects, however, remain incompletely understood. Monomethylarsonous acid [MMA(III)] is a highly toxic and stable metabolite of inorganic arsenic. To exploit the mechanisms through which MMA(III) exerts its cytotoxic effect, we adopted a quantitative proteomic approach, by coupling stable isotope labeling by amino acids in cell culture (SILAC) with LC-MS/MS analysis, to examine the variation in the entire proteome of GM00637 human skin fibroblasts following acute MMA(III) exposure. Among the ~6500 unique proteins quantified, ~300 displayed significant changes in expression after exposure with 2?M MMA(III) for 24h. Subsequent analysis revealed the perturbation of de novo cholesterol biosynthesis, selenoprotein synthesis and Nrf2 pathways evoked by MMA(III) exposure. Particularly, MMA(III) treatment resulted in considerable down-regulation of several enzymes involved in cholesterol biosynthesis. In addition, real-time PCR analysis showed reduced mRNA levels of select genes in this pathway. Furthermore, MMA(III) exposure contributed to a distinct decline in cellular cholesterol content and significant growth inhibition of multiple cell lines, both of which could be restored by supplementation of cholesterol to the culture media. Collectively, the present study demonstrated that the cytotoxicity of MMA(III) may arise, at least in part, from the down-regulation of cholesterol biosynthesis enzymes and the resultant decrease of cellular cholesterol content. PMID:24625837

Guo, Lei; Xiao, Yongsheng; Wang, Yinsheng

2014-05-15

12

Degradation of type IV collagen by neoplastic human skin fibroblasts  

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An assay for the degradation of type IV (basement membrane) collagen was developed as a biochemical marker for neoplastic cells from chemically transformed human skin fibroblasts. Type IV collagen was isolated from basement membrane of Syrian hamster lung and type I collagen was isolated from rat tails; the collagens were radioactively labelled by reductive alkylation. The abilities of normal (KD) and chemically transformed (Hut-11A) human skin fibroblasts to degrade the collagens were studied. A cell-associated assay was performed by growing either normal or transformed cells in the presence of radioactively labelled type IV collagen and measuring the released soluble peptides in the medium. This assay also demonstrated that KD cells failed to synthesize an activity capable of degrading type IV collagen whereas Hut-11A cells degraded type IV collagen in a linear manner for up to 4 h. Human serum at very low concentrations, EDTA and L-cysteine inhibited the enzyme activity, whereas protease inhibitors like phenylmethyl sulfonyl fluoride, N-ethyl maleimide or soybean trypsin inhibitor did not inhibit the enzyme from Hut-11A cells. These results suggest that the ability to degrade specifically type IV collagen may be an important marker for neoplastic human fibroblasts and supports a role for this collagenase in tumor cell invasion

1985-01-01

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Respiratory chain analysis of skin fibroblasts in mitochondrial disease.  

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NADH:ubiquinone dehydrogenase (complex I) deficiency can be diagnosed from cultured skin fibroblasts using a number of methods, the most commonly used is a linked assay of rotenone-sensitive complex I + III activity (NADH:cytochrome c reductase). Because of interference from diaphorases, this method requires either the isolation of mitochondria (or at least partial purification). For a suitable mitochondrial preparation from skin fibroblasts, this requires the culturing of 4-20 individual 100mm tissue culture plates, depending on the purity of preparation required. These assays are therefore time-consuming, and do not assist in a rapid diagnosis. There is also no clear demarkation between the normal range of activity and the deficient range since mild mutations can produce only partial decreases in complex I activity. Equally, assaying patient cells that do not have a specific deficiency may prove to be time-wasting in the process of providing a quick, definitive clinical diagnosis. The lactate/pyruvate ratio of fibroblasts has been used to indicate the extent of respiratory chain involvement, as cells with a metabolic defect usually produce more lactate with an increased ratio from 25:1 to much higher values [Methods Enzymol. 264 (1996) 454]. This measurement may not always be conclusive, as the values can fluctuate as a result of culture medium, cell passage number, cell number and viability. In this report, we evaluate the use of pyruvate oxidation measurements from whole cells prepared from a single plate of cultured fibroblasts as an alternative to lactate/pyruvate ratios, or other methods both direct and indirect as indicators of the extent of respiratory chain involvement and the possibility of a defect within complex I. Whole cell 2-14C pyruvate oxidation appears to indicate the presence of a complex I defect in patients compared to normal controls more reliably than L/P ratios, but this has some puzzling exceptions. PMID:16120400

Cameron, Jessie M; Levandovskiy, Valeriy; MacKay, Neviana; Robinson, Brian H

2004-09-01

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The Nf1 Tumor Suppressor Regulates Mouse Skin Wound Healing, Fibroblast Proliferation, and Collagen Deposited by Fibroblasts  

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Neurofibromatosis type 1 patients develop peripheral nerve tumors (neurofibromas) composed mainly of Schwann cells and fibroblasts, in an abundant collagen matrix produced by fibroblasts. Trauma has been proposed to trigger neurofibroma formation. To test if loss of the neurofibromatosis type 1 gene (Nf1) compromises fibroblast function in vivo following trauma, skin wounding was performed in Nf1 knockout mice. The pattern and amount of collagen-rich granulation bed tissue, manufactured by fi...

Atit, Radhika P.; Crowe, Maria J.; Greenhalgh, David G.; Wenstrup, Richard J.; Ratner, Nancy

1999-01-01

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Fibroblast-mediated contraction in actinically exposed and actinically protected aging skin  

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The changes in skin morphology over time are a consequence of both chronologic aging and the accumulation of environmental exposure. Through observation, we know that actinic radiation intensifies the apparent aging of skin. We have investigated the effects of aging and actinic radiation on the ability of fibroblasts to contract collagen-fibroblast lattices. Preauricular and postauricular skin samples were obtained from eight patients aged 49 to 74 undergoing rhytidectomy. The samples were kept separate, and the fibroblasts were grown in culture. Lattices constructed with preauricular fibroblasts consistently contracted more than lattices containing postauricular fibroblasts. The difference in amount of contraction in 7 days between sites was greatest for the younger patients and decreased linearly as donor age increased (r = -0.96). This difference may be due to preauricular fibroblasts losing their ability to contract a lattice as aging skin is exposed to more actinic radiation

1990-01-01

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FSP1+ Fibroblasts Promote Skin Carcinogenesis by Maintaining MCP-1-Mediated Macrophage Infiltration and Chronic Inflammation  

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Cancer development is often associated with increased fibroblast proliferation and extensive fibrosis; however, the role of fibroblasts during carcinogenesis remains largely unknown. Using the 7,12-dimethylbenz-(a)anthracene and 12-O-tetradecanoylphorbol-13-acetate–induced two-stage skin carcinogenesis model, we demonstrated here that there was a massive accumulation and proliferation of fibroblasts in the skin shortly after application of carcinogen. Selective abatement of these cells duri...

Zhang, Jinhua; Chen, Lin; Xiao, Mingjie; Wang, Chunhui; Qin, Zhihai

2011-01-01

17

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

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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

18

The Level and Stability of Residual Catalase in Cultured Acatalasemic Skin Fibroblasts  

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Full Text Available In an attempt to determine the level and heat stability of residual catalase in somatic cells of acatalasemic Japanese, skin fibroblasts from an acatalasemic subject were cultured, and the catalase activity of the cultured fibroblasts was compared with that of cultured normal fibroblasts. Catalase activity was determined using an oxygen electrode. The residual catalase activity in cultured acatalasemic fibroblasts was 10% of the normal. The heat stability at 55 degrees C of residual catalase in the acatalasemic fibroblasts was similar to that of normal fibroblasts.

Ogata,Masana

1987-10-01

19

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

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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

20

A Marfan syndrome gene expression phenotype in cultured skin fibroblasts  

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Full Text Available Abstract Background Marfan syndrome (MFS is a heritable connective tissue disorder caused by mutations in the fibrillin-1 gene. This syndrome constitutes a significant identifiable subtype of aortic aneurysmal disease, accounting for over 5% of ascending and thoracic aortic aneurysms. Results We used spotted membrane DNA macroarrays to identify genes whose altered expression levels may contribute to the phenotype of the disease. Our analysis of 4132 genes identified a subset with significant expression differences between skin fibroblast cultures from unaffected controls versus cultures from affected individuals with known fibrillin-1 mutations. Subsequently, 10 genes were chosen for validation by quantitative RT-PCR. Conclusion Differential expression of many of the validated genes was associated with MFS samples when an additional group of unaffected and MFS affected subjects were analyzed (p-value -6 under the null hypothesis that expression levels in cultured fibroblasts are unaffected by MFS status. An unexpected observation was the range of individual gene expression. In unaffected control subjects, expression ranges exceeding 10 fold were seen in many of the genes selected for qRT-PCR validation. The variation in expression in the MFS affected subjects was even greater.

Emond Mary

2007-09-01

 
 
 
 
21

Transplantation of Composite Skin Containing Keratinocytes Cultured on a Fibroblast-conditioned Acellular Dermal Matrix  

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To evaluate the role of fibroblasts in composite skin reconstructed in vitro, four different types of composite skin (A, B, C, and D) were prepared. Human keratinocytes were seeded onto the epidermal side of an acellular dermal matrix (ADM) in type A. Keratinocytes were seeded onto the epidermal side of an ADM and human fibroblasts onto the dermal side in type B. Both keratinocytes and fibroblasts were seeded onto the epidermal side in type C. Type D consisted of fibroblasts on both sides of ...

Xiao, S. -c; Ben, D. -f; Yang, J.; Tang, H. -t; Wang, G. -q; Yang, Y.; Yu, W. -r; Xia, Z. -f

2005-01-01

22

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  

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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

23

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

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Total collagen synthesis is decreased by about 29% (P 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

24

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

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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

1995-11-01

25

Study of superoxide dismutase mechanisms of action on fibrosis myo-fibroblasts cultured in a reconstituted skin model  

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Fibrosis of the skin is frequently observed after therapeutic and accidental irradiations. In order to better understand the mechanisms leading to skin fibrosis, we tried to characterize the differences between normal and fibrotic fibroblasts isolated from pig skin. (authors)

Vozenin, M.C.; Lefaix, J.L.; Martin, M. [Laboratoire de Radiologie Appliquee, CEA Saclay, 91 - Gif-sur-Yvette (France)

1997-03-01

26

AGE-RELATED GENE EXPRESSION CHANGES IN HUMAN SKIN FIBROBLASTS INDUCED BY MMS  

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Age-Related Gene Expression Changes In Human Skin Fibroblasts Induced By methyl methanesulfonate. Geremy W. Knapp, Alan H. Tennant, and Russell D. Owen. Environmental Carcinogenesis Division, National Health and Environmental Effects Research Laboratory, U. S. Environmental Prote...

27

Coxsackievirus B3 infection alters plasma membrane of neonatal skin fibroblasts.  

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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

28

Effects of Cigarette Smoke Extracts on the Growth and Senescence of Skin Fibroblasts In Vitro  

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Epidemiological studies have shown that cigarette smoke (CS), a very common environmental factor, plays an important role in skin aging. Although some in vivo studies have suggested that CS affects skin aging, the detailed effects of CS on skin cells in vitro remain largely unknown. In this study, we investigated the effects of cigarette smoke extract (CSE) on the growth, proliferation, and senescene of skin fibroblasts and the possible mechanism underlying these effects. Primary cultured hum...

Yang, Gao-yun; Zhang, Chun-lei; Liu, Xiang-chen; Qian, Ge; Deng, Dan-qi

2013-01-01

29

In vitro culture of skin fibroblast cells for potential cloning by nuclear transfer  

International Nuclear Information System (INIS)

Donor cell lines were developed from skin tissue for the conservation of the endangered Jaiselmeri camel breed of India. Average cell proliferation rates varied from 0.82 to 0.69 in different passages, and population doubling time from 29.3 h to 34.8 h. Around 15 population doublings were accomplished during this culturing. Cell viability was 97 to 99% in different passages. Growth curves of cells from the JC-5 cell line reached a plateau on day 7, while the slower-growing cultures of JC-3 showed elevation even on day 10, possibly due to donor age differences. Cell proliferation rates by both cell count and MTT absorbance showed similar patterns, with a correlation coefficient of 0.79. MTT assay, a colorimetric method, can handle large samples in somatic cell cultures. Diploid chromosomal counts in passages 1, 3 and 5 were normal (2N=74, XY) in 97% of the cells. Occasional metaphase plates showed polyploidy. The present baseline data on standard growth curve, linear relationship in colorimetric assay for estimation of cell proliferation rate, and normal ploidy and karyological levels in camel skin fibroblast cells in multiplication could be useful in developing competent donor somatic cell lines for conservation now and revival of this camel breed by cloning in the future. (author)

2003-10-06

30

Collagen fragmentation promotes oxidative stress and elevates matrix metalloproteinase-1 in fibroblasts in aged human skin.  

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Aged human skin is fragile because of fragmentation and loss of type I collagen fibrils, which confer strength and resiliency. We report here that dermal fibroblasts express increased levels of collagen-degrading matrix metalloproteinases-1 (MMP-1) in aged (>80 years old) compared with young (21 to 30 years old) human skin in vivo. Transcription factor AP-1 and alpha2beta1 integrin, which are key regulators of MMP-1 expression, are also elevated in fibroblasts in aged human skin in vivo. MMP-1 treatment of young skin in organ culture causes fragmentation of collagen fibrils and reduces fibroblast stretch, consistent with reduced mechanical tension, as observed in aged human skin. Limited fragmentation of three-dimensional collagen lattices with exogenous MMP-1 also reduces fibroblast stretch and mechanical tension. Furthermore, fibroblasts cultured in fragmented collagen lattices express elevated levels of MMP-1, AP-1, and alpha2beta1 integrin. Importantly, culture in fragmented collagen raises intracellular oxidant levels and treatment with antioxidant MitoQ(10) significantly reduces MMP-1 expression. These data identify positive feedback regulation that couples age-dependent MMP-1-catalyzed collagen fragmentation and oxidative stress. We propose that this self perpetuating cycle promotes human skin aging. These data extend the current understanding of the oxidative theory of aging beyond a cellular-centric view to include extracellular matrix and the critical role that connective tissue microenvironment plays in the biology of aging. PMID:19116368

Fisher, Gary J; Quan, Taihao; Purohit, Trupta; Shao, Yuan; Cho, Moon Kyun; He, Tianyuan; Varani, James; Kang, Sewon; Voorhees, John J

2009-01-01

31

Collagen Fragmentation Promotes Oxidative Stress and Elevates Matrix Metalloproteinase-1 in Fibroblasts in Aged Human Skin  

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Aged human skin is fragile because of fragmentation and loss of type I collagen fibrils, which confer strength and resiliency. We report here that dermal fibroblasts express increased levels of collagen-degrading matrix metalloproteinases-1 (MMP-1) in aged (>80 years old) compared with young (21 to 30 years old) human skin in vivo. Transcription factor AP-1 and ?2?1 integrin, which are key regulators of MMP-1 expression, are also elevated in fibroblasts in aged human skin in vivo. MMP-1 tre...

2009-01-01

32

Defective mitochondrial fusion, altered respiratory function, and distorted cristae structure in skin fibroblasts with heterozygous OPA1 mutations.  

Science.gov (United States)

Deleterious consequences of heterozygous OPA1 mutations responsible for autosomal dominant optic atrophy remain a matter of debate. Primary skin fibroblasts derived from patients have shown diverse mitochondrial alterations that were however difficult to resolve in a unifying scheme. To address the potential use of these cells as disease model, we undertook parallel and quantitative analyses of the diverse reported alterations in four fibroblast lines harboring different OPA1 mutations, nonsense or missense, in the guanosine triphosphatase or the C-terminal coiled-coil domains. We tackled several factors potentially underlying discordant reports and showed that fibroblasts with heterozygous OPA1 mutations present with several mitochondrial alterations. These included defective mitochondrial fusion during pharmacological challenge with the protonophore carbonyl cyanide m-chlorophenyl hydrazone, significant mitochondrial elongation with decreased OPA1 and DRP1 proteins, and abnormal mitochondrial fragmentation during glycolysis shortage or exogenous oxidative stress. Respiratory complex IV activity and subunits steady-state were decreased without alteration of the mitochondrial deoxyribonucleic acid size, amount or transcription. Physical link between OPA1 protein and oxidative phosphorylation was shown by reciprocal immunoprecipitation. Altered cristae structure coexisted with normal response to pro-apoptotic stimuli and expression of Bax or Bcl2 proteins. Skin fibroblasts with heterozygous OPA1 mutations thus share significant mitochondrial remodeling, and may therefore be useful for analyzing disease pathophysiology. Identifying whether the observed alterations are also present in ganglion retinal cells, and which of them underlies their degeneration process remains however an essential goal for therapeutic strategy. PMID:22800932

Agier, Virginie; Oliviero, Patricia; Lainé, Jeanne; L'Hermitte-Stead, Caroline; Girard, Samantha; Fillaut, Sandrine; Jardel, Claude; Bouillaud, Frédéric; Bulteau, Anne Laure; Lombès, Anne

2012-10-01

33

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 patients

2007-06-01

34

Hydroxyeicosatetraenoic acid metabolism in cultured human skin fibroblasts. Evidence for peroxisomal beta-oxidation.  

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To determine whether the peroxisome is responsible for hydroxyeicosatetraenoic acid (HETE) oxidation, 12- and 15-HETE oxidation was measured in normal and peroxisomal deficient skin fibroblasts from patients with Zellweger's (cerebrohepatorenal) syndrome. When incubated for 1 h with normal fibroblasts, reverse phase HPLC indicated that 24% of the 12-HETE radioactivity was converted to one major polar metabolite. Chemical derivatization followed by reverse phase HPLC and TLC indicated that thi...

Gordon, J. A.; Figard, P. H.; Spector, A. A.

1990-01-01

35

Proteomic Identification of Cathepsin B and Nucleophosmin as Novel UVA-Targets in Human Skin Fibroblasts  

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Solar UVA exposure plays a causative role in skin photoaging and photocarcinogenesis. Here we describe the proteomic identification of novel UVA-targets in human dermal fibroblasts following a 2D-DIGE (two-dimensional-difference-gel-electrophoresis) approach. Fibroblasts were exposed to non-cytotoxic doses of UVA or left untreated, and total protein extracts underwent CyDye-labeling followed by 2D-DIGE/mass spectrometric identification of differentially expressed proteins, confirmed independe...

Lamore, Sarah D.; Qiao, Shuxi; Horn, David; Wondrak, Georg T.

2010-01-01

36

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

1981-01-01

37

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

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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

38

Transforming growth factor-beta reverses a posttranscriptional defect in elastin synthesis in a cutis laxa skin fibroblast strain.  

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Skin fibroblasts from two cases of autosomal recessive cutis laxa (CL), having insignificant elastin production and mRNA levels, were challenged with transforming growth factor beta-1 (TGF-beta 1). Elastin production was brought from undetectable values to amounts typical of normal human skin fibroblasts in a dose-dependent fashion. Basic fibroblast growth factor (100 ng/ml) alone or in combination with TGF-beta 1 reduced elastin production and mRNA expression in CL skin fibroblasts more exte...

Zhang, M. C.; Giro, M.; Quaglino, D.; Davidson, J. M.

1995-01-01

39

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

40

Mesenchymal–epithelial interactions in the skin: increased expression of dickkopf1 by palmoplantar fibroblasts inhibits melanocyte growth and differentiation  

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We investigated whether or not the topographic regulation of melanocyte differentiation is determined by mesenchymal–epithelial interactions via fibroblast-derived factors. The melanocyte density in palmoplantar human skin (i.e., skin on the palms and the soles) is five times lower than that found in nonpalmoplantar sites. Palmoplantar fibroblasts significantly suppressed the growth and pigmentation of melanocytes compared with nonpalmoplantar fibroblasts. Using cDNA microarray analysis, fi...

Yamaguchi, Yuji; Itami, Satoshi; Watabe, Hidenori; Yasumoto, Ken-ichi; Abdel-malek, Zalfa A.; Kubo, Tateki; Rouzaud, Franc?ois; Tanemura, Atsushi; Yoshikawa, Kunihiko; Hearing, Vincent J.

2004-01-01

 
 
 
 
41

Enhanced biosynthesis of human skin collagenase in fibroblast cultures from recessive dystrophic epidermolysis bullosa.  

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Using a sensitive, specific immunoprecipitation method, the biosynthesis of human skin collagenase was studied in fibroblast cultures from patients with recessive dystrophic epidermolysis bullosa. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of solubilized immunoprecipitates showed two 3H-labeled procollagenase species that comigrated with those harvested from control cultures. Recessive dystrophic epidermolysis bullosa cultures accumulated increased amounts of collagenase. Both ...

Valle, K. J.; Bauer, E. A.

1980-01-01

42

Assessment of the potential skin irritation of lysine-derivative anionic surfactants using mouse fibroblast and human keratinocytes as an alternative to animal testing  

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Purpose. The aim of this study was to identify new surfactants with low skin irritant properties for use in pharmaceutical and cosmetic formulations, employing cell culture as an alternative method to in vivo testing. In addition, we sought to establish whether potential cytotoxic properties were related to the size of the counterions bound to the surfactants. Methods. Cytotoxicity was assessed in the mouse fibroblast cell line 3T6, and the human keratinocyte cell line NCTC 2544, using the MT...

Sa?nchez Molina, Lourdes; Mitjans Arnal, Montserrat; Infante Marti?nez-pardo, Ma Rosa; Vinardell Marti?nez-hidalgo, Ma Pilar

2004-01-01

43

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

44

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

45

Phenotypic correction of factor IX deficiency in skin fibroblasts of hemophilic dogs.  

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Primary skin fibroblasts from hemophilic dogs were transduced by recombinant retrovirus (LNCdF9L) containing a canine factor IX cDNA. High levels of biologically active canine factor IX (1.0 micrograms per 10(6) cells per 24 hr) were secreted in the medium. The level of factor IX produced increased substantially if the cells were stimulated by basic fibroblast growth factor during infection. Additionally, we also report that endothelial cells transduced by this virus can produce high levels o...

Axelrod, J. H.; Read, M. S.; Brinkhous, K. M.; Verma, I. M.

1990-01-01

46

DNA-protein crosslinking in normal human skin fibroblasts exposed to solar ultraviolet wavelengths  

International Nuclear Information System (INIS)

Three normal human skin fibroblast cell lines were exposed to the simulated solar UV radiation produced by a fluorescent sunlamp (wavelength components shorter than either 295, 305 or 315 nm were excluded). The level of DNA-protein crosslinks (DPC) was then measured in those cells either immediately after irradiation or following a 24 h incubation. Cells were exposed to fluences that induce similar levels of DPC. For cells exposed to 10 kJ m"-"2 of sunlamp UV>295 nm, the level of DPC exhibited a 2-5-fold increase following incubation. In contrast, 40-100% of the DPC were removed upon incubation of cells irradiated with either 10 kJ m"-"2 of sunlamp UV>305 nm or 150 kJ m"-"2 of sunlamp UV>315 nm. A major difference between the effects induced by these wavelength regions is that, in addition to DPC, a very high level of pyrimidine dimers is also produced by sunlamp UV>295 nm, whereas much lower dimer yields result from treatment with either sunlamp UV>305 nm or sunlamp UV>315 nm. A potential role for type II DNA topoisomerase in the formation of these DPC resulting from either the change in conformational structure caused by the presence of a high level of dimers or an involvement of this enzyme in dimer excision repair is discussed. (author)

1987-01-01

47

Enhancing structural support of the dermal microenvironment activates fibroblasts, endothelial cells and keratinocytes in aged human skin in vivo  

Science.gov (United States)

The dermal extracellular matrix (ECM) provides strength and resiliency to skin. The ECM consists mostly of type I collagen fibrils, which are produced by fibroblasts. Binding of fibroblasts to collagen fibrils generates mechanical forces, which regulate cellular morphology and function. With aging, collagen fragmentation reduces fibroblast-ECM binding and mechanical forces, resulting in fibroblast shrinkage and reduced function including collagen production. Here, we report that these age-related alterations are largely reversed by enhancing structural support of the ECM. Injection of dermal filler, cross-linked hyaluronic acid, into the skin of persons over seventy years-old stimulates fibroblasts to produce type I collagen. This stimulation is associated with localized increased of mechanical forces, indicated by fibroblast elongation/spreading, and mediated by up-regulation of type II TGF-? receptor and connective tissue growth factor. Interestingly, enhanced mechanical support of the ECM also stimulates fibroblast proliferation, expands vasculature, and increases epidermal thickness. Consistent with our observations in human skin, injection of filler into dermal equivalent cultures causes elongation of fibroblasts, coupled with type I collagen synthesis, which is dependent on the TGF-? signaling pathway. Thus, fibroblasts in aged human skin retain their capacity for functional activation, which is restored by enhancing structural support of the ECM.

Quan, Taihao; Wang, Frank; Shao, Yuan; Rittie, Laure; Xia, Wei; Orringer, Jeffrey S.; Voorhees, John J.; Fisher, Gary J.

2012-01-01

48

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

49

Cultured skin fibroblasts from patients with porokeratosis are hypersensitive to the lethal effects of X-radiation  

Energy Technology Data Exchange (ETDEWEB)

Porokeratosis is an autosomal dominant inherited skin disorder. The lesions are characterized by localized abnormal keratinization and may develop into malignant tumors. To determine the cellular basis of the cancer susceptibility associated with this skin condition, we examined the colony-forming ability of X-ray or ultraviolet (UV) light irradiated, cultured fibroblasts derived from porokeratosis patients' normal-appearing skin. Four fibroblast strains derived from four porokeratosis patients' skin were significantly hypersensitive to the lethal effects of X-radiation. However, they all showed a similar sensitivity to strains from normal donors to 254 nm UV light. The hypersensitivity to X-ray radiation in cultured skin fibroblasts from porokeratosis patients suggests an inherent instability of cellular DNA and may prbably be associated with the cancer-prone nature of this skin condition.

Otsuka, Fujio; Watanabe, Ryohji; Moro, Akiko; Ohkochi, Hitoshi; Ishibashi, Yasumasa

1989-01-01

50

Cultured skin fibroblasts from patients with porokeratosis are hypersensitive to the lethal effects of X-radiation  

International Nuclear Information System (INIS)

Porokeratosis is an autosomal dominant inherited skin disorder. The lesions are characterized by localized abnormal keratinization and may develop into malignant tumors. To determine the cellular basis of the cancer susceptibility associated with this skin condition, we examined the colony-forming ability of X-ray or ultraviolet (UV) light irradiated, cultured fibroblasts derived from porokeratosis patients' normal-appearing skin. Four fibroblast strains derived from four porokeratosis patients' skin were significantly hypersensitive to the lethal effects of X-radiation. However, they all showed a similar sensitivity to strains from normal donors to 254 nm UV light. The hypersensitivity to X-ray radiation in cultured skin fibroblasts from porokeratosis patients suggests an inherent instability of cellular DNA and may prbably be associated with the cancer-prone nature of this skin condition. (author)

1989-01-01

51

Culturing of skin fibroblasts in a thin PLGA-collagen hybrid mesh.  

Science.gov (United States)

A thin biodegradable hybrid mesh of synthetic poly(DL-lactic-co-glycolic acid) (PLGA) and naturally derived collagen was used for three-dimensional culture of human skin fibroblasts. The hybrid mesh was constructed by forming web-like collagen microsponges in the openings of a PLGA knitted mesh. The behaviors of the fibroblasts on the hybrid mesh and PLGA knitted mesh were compared. The efficiency of cell seeding was much higher and the cells grew more quickly in the hybrid mesh than in the PLGA mesh. The fibroblasts in the PLGA mesh grew from the peripheral PLGA fibers toward the centers of the openings, while those in the hybrid mesh also grew from the collagen microsponges in the openings of the mesh resulting in a more homogenous growth. The proliferated cells and secreted extracellular matrices were more uniformly distributed in the hybrid mesh than in the PLGA mesh. Histological staining of in vitro cultured fibroblast/mesh implants indicated that the fibroblasts were distributed throughout the hybrid mesh and formed a uniform layer of dermal tissue having almost the same thickness as that of the hybrid mesh. However, the tissue formed in the PLGA mesh was thick adjacent to the PLGA fibers and thin in the center of the openings. Fibroblasts cultured in the hybrid mesh were implanted in the back of nude mouse. Dermal tissues were formed after 2 weeks and became epithelialized after 4 weeks. The results indicate that the web-like collagen microsponges formed in the openings of the PLGA knitted mesh increased the efficiency of cell seeding, improved cell distribution, and therefore facilitated rapid formation of dermal tissue having a uniform thickness. PLGA-collagen hybrid mesh may be useful for skin tissue engineering. PMID:15585258

Chen, Guoping; Sato, Takashi; Ohgushi, Hajime; Ushida, Takashi; Tateishi, Tetsuya; Tanaka, Junzo

2005-05-01

52

Protective effect of maghemite nanoparticles on ultraviolet-induced photo-damage in human skin fibroblasts  

Science.gov (United States)

This study examined the optical properties of an oxidized form of maghemite (?-Fe2O3) nanoparticles and their protective effects against the photoaging of human skin fibroblasts irradiated with ultraviolet (UV) light. Nanoparticles with diameters ranging from 8.7 to 12 nm were prepared using a chemical co-precipitation method. The nanoparticles were coated with two surfactants to obtain a water-based product. The onset of the absorption of the ?-Fe2O3 nanoparticles in the UV-visible absorption spectra increased with increasing particle size. The ?-Fe2O3 nanoparticles significantly inhibited the production of matrix metalloproteinase-1 in human skin fibroblast HS 68 cells by 60% compared with the UV-irradiated control. These results suggest that ?-Fe2O3 nanoparticles have photoprotective properties, and have potential use as an agent against photoaging.

Lee, Kwon-Jai; An, Jeung-Hee; Shin, Jae-Soo; Kim, Dong-Hee; Kim, Changman; Ozaki, Hajime; Koh, Jae-Gui

2007-11-01

53

Protective effect of maghemite nanoparticles on ultraviolet-induced photo-damage in human skin fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

This study examined the optical properties of an oxidized form of maghemite ({gamma}-Fe{sub 2}O{sub 3}) nanoparticles and their protective effects against the photoaging of human skin fibroblasts irradiated with ultraviolet (UV) light. Nanoparticles with diameters ranging from 8.7 to 12 nm were prepared using a chemical co-precipitation method. The nanoparticles were coated with two surfactants to obtain a water-based product. The onset of the absorption of the {gamma}-Fe{sub 2}O{sub 3} nanoparticles in the UV-visible absorption spectra increased with increasing particle size. The {gamma}-Fe{sub 2}O{sub 3} nanoparticles significantly inhibited the production of matrix metalloproteinase-1 in human skin fibroblast HS 68 cells by 60% compared with the UV-irradiated control. These results suggest that {gamma}-Fe{sub 2}O{sub 3} nanoparticles have photoprotective properties, and have potential use as an agent against photoaging.

Lee, Kwon-Jai [Department of Physics, Soongsil University, Seoul 156-743 (Korea, Republic of); An, Jeung-Hee [Brain Disease Research Center, Ajou University School of Medicine, Suwon 443-749 (Korea, Republic of); Shin, Jae-Soo [Department of Advanced Materials Engineering, Daejeon University, Daejeon 300-716 (Korea, Republic of); Kim, Dong-Hee [Department of Pathology Lab of Oriental Medicine College, Daejeon University, Daejeon 300-716 (Korea, Republic of); Kim, Changman [Department of Electrical Engineering and Bioscience, Waseda University, Tokyo 169-8555 (Japan); Ozaki, Hajime [Department of Electrical Engineering and Bioscience, Waseda University, Tokyo 169-8555 (Japan); Koh, Jae-Gui [Department of Physics, Soongsil University, Seoul 156-743 (Korea, Republic of)

2007-11-21

54

Protective effect of maghemite nanoparticles on ultraviolet-induced photo-damage in human skin fibroblasts  

International Nuclear Information System (INIS)

This study examined the optical properties of an oxidized form of maghemite (?-Fe2O3) nanoparticles and their protective effects against the photoaging of human skin fibroblasts irradiated with ultraviolet (UV) light. Nanoparticles with diameters ranging from 8.7 to 12 nm were prepared using a chemical co-precipitation method. The nanoparticles were coated with two surfactants to obtain a water-based product. The onset of the absorption of the ?-Fe2O3 nanoparticles in the UV-visible absorption spectra increased with increasing particle size. The ?-Fe2O3 nanoparticles significantly inhibited the production of matrix metalloproteinase-1 in human skin fibroblast HS 68 cells by 60% compared with the UV-irradiated control. These results suggest that ?-Fe2O3 nanoparticles have photoprotective properties, and have potential use as an agent against photoaging

2007-11-21

55

A case of actinic prurigo showing hypersensitivity of skin fibroblasts to ultraviolet A (UVA).  

Science.gov (United States)

We here report a patient with actinic prurigo. He had had erythematous papulovesicular eruptions on the sun-exposed sites from fall to early summer for 4 years. The lesions healed leaving atrophic scars. The histology showed epidermal necrosis and dermal dense perivascular lymphohistiocytic infiltration and edema. His minimal erythema doses to ultraviolet B (UVB) and UVA were normal and lowered, respectively. Skin lesions were produced by repeated irradiation with UVA plus UVB, but not with UVA alone. Then he was diagnosed as having actinic prurigo. Skin fibroblasts from the patient were hypersensitive to UVA. We believe that the hypersensitivity relates to the pathomechanisms of the photosensitivity in the case. UVA sensitivity of fibroblasts may be useful for differentiating actinic prurigo, hydroa vacciniforme, and other similar photosensitive disorders. PMID:10721864

Kuno, Y; Sato, K; Hasegawa, K; Tsuji, T

2000-02-01

56

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

1981-01-01

57

Transient in vitro epigenetic reprogramming of skin fibroblasts into multipotent cells  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Multipotent stem cells have the potential to establish a new field of promising regenerative medicine to treat tissue damage, genetic disorders, and degenerative diseases. However, limited resource of stem cells has turned to be an evitable obstacle in clinical applications. We utilized a simple in vitro epigenetic reprogramming approach to convert skin fibroblasts into multipotent cells. After transient reprogramming, stem cell markers, including Oct4 and Nanog, became activated in the treat...

2010-01-01

58

Oxidant stress leads to transcriptional activation of the human heme oxygenase gene in cultured skin fibroblasts.  

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Treatment of cultured human skin fibroblasts with near-UV radiation, hydrogen peroxide, and sodium arsenite induces accumulation of heme oxygenase mRNA and protein. In this study, these treatments led to a dramatic increase in the rate of RNA transcription from the heme oxygenase gene but had no effect on mRNA stability. Transcriptional activation, therefore, appears to be the major mechanism of stimulation of expression of this gene by either oxidative stress or sulfydryl reagents.

Keyse, S. M.; Applegate, L. A.; Tromvoukis, Y.; Tyrrell, R. M.

1990-01-01

59

Recovery from UV-induced potentially lethal damage in systemic lupus erythematosus skin fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

The repair of ultraviolet light-induced potentially lethal damage was investigated in density-inhibited skin fibroblast cell strains derived from patients with systemic lupus erythematosus. The effect of exposure to polychromatic ultraviolet light composed of environmentally relevant wavelengths or to the more commonly studied, short wavelength (254 nm) ultraviolet light was studied. Systemic lupus erythematosus cells, which are hypersensitive to ultraviolet light under growth promoting conditions, were able to repair potentially lethal damage as well as normal cells.

Zamansky, G.B.

1986-08-01

60

Recovery from UV-induced potentially lethal damage in systemic lupus erythematosus skin fibroblasts  

International Nuclear Information System (INIS)

The repair of ultraviolet light-induced potentially lethal damage was investigated in density-inhibited skin fibroblast cell strains derived from patients with systemic lupus erythematosus. The effect of exposure to polychromatic ultraviolet light composed of environmentally relevant wavelengths or to the more commonly studied, short wavelength (254 nm) ultraviolet light was studied. Systemic lupus erythematosus cells, which are hypersensitive to ultraviolet light under growth promoting conditions, were able to repair potentially lethal damage as well as normal cells. (author)

1986-01-01

 
 
 
 
61

Lysinuric protein intolerance mutation is expressed in the plasma membrane of cultured skin fibroblasts.  

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Lysinuric protein intolerance (LPI) is an autosomal recessive phenotype consistent with impaired transport of cationic amino acids at the basolateral membrane of intestinal and renal epithelia. On the assumption that the basolateral membrane of epithelial cells and plasma membrane of parenchymal cells are functional analogues, we studied transport of cationic amino acids by cultured skin fibroblasts from LPI and control subjects matched for age, sex, and site of biopsy. We measured Na+-indepe...

1987-01-01

62

Estrogen Prevents Oxidative Damage to the Mitochondria in Friedreich's Ataxia Skin Fibroblasts  

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Estrogen and estrogen-related compounds have been shown to have very potent cytoprotective properties in a wide range of disease models, including an in vitro model of Friedreich's ataxia (FRDA). This study describes a potential estrogen receptor (ER)-independent mechanism by which estrogens act to protect human FRDA skin fibroblasts from a BSO-induced oxidative insult resulting from inhibition of de novo glutathione (GSH) synthesis. We demonstrate that phenolic estrogens, independent of any ...

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

2012-01-01

63

Radioprotective effect of c-ski on rat skin fibroblast in vitro  

International Nuclear Information System (INIS)

Objective: To examine radioprotective effect of c-ski on rat skin fibroblast in vitro and explore its possible mechanism. Methods: The effect of soft X-ray irradiation at dose varied from 2 to 8 Gy on cell apoptosis in rat skin fibroblast were determined by flow cytometry with Annexin-V-FITC-PI labelling. The effect of c-ski gene transfection on cell apoptosis was evaluated after soft X-ray irradiation of 4 Gy. The protein expressions of Bax and Bcl-2 after c-ski gene transfection were measured with the Western blot method. Results: Soft X-ray irradiation increases cell apoptosis, and the increase is proportional to the irradiation dose. Apoptosis ratio increases with time since the irradiation, and reaches its peak at 36h after the irradiation, c-ski gene was observed to markedly decrease apoptosis index at 24 h after soft X-ray irradiation of 4 Gy compared to the control group, significant increase of the protein expression of Bcl-2 was observed. C-ski gene was found no significant effect on the protein expression of Bax. Conclusion: c-ski gene can decrease radiation sensitivity of skin fibroblast, promoting Bcl-2 protein expression is one of its possible mechanism for this radioprotective effects. (authors)

2006-04-01

64

Heterogeneous response to X-ray and ultraviolet light irradiations of cultured skin fibroblasts in two families with Gardner's Syndrome  

International Nuclear Information System (INIS)

A heterogeneous response to X-ray and far UV (254 nm) light irradiations was found in cultured skin fibroblast lines from 2 separate families with Gardner's syndrome. When compared to 2 normal control cultures and cultures from 2 patients with nonfamilial colon cancer, cultures from 4 clinically affected members of family 1 showed increased sensitivity to the lethal effects of both X-ray and UV light irradiations. These cells also showed a delayed pattern of X-ray potentially lethal damage repair (PLDR) and absent UV PLDR. In contrast, cultures from 3 members of family 2 (2 of whom were clinically affected) showed a normal response of survival and PLDR to both X-ray and UV light irradiations. Thus increased sensitivity of cultured skin fibroblasts to X-ray and UV light irradiations was not a consistent in vitro finding in patients with Gardner's syndrome. However, in families with Gardner's syndrome who demonstrate in vitro radiosensitivity, additional studies are needed to assess the usefulness of these techniques in detecting affected individuals prior to the development of colon carcinoma and other manifestations

1982-01-01

65

Radiosensitivity of fibroblasts obtained from a cafe-au-lait spot and normal-appearing skin of a patient with neurofibromatosis (NF-6)  

International Nuclear Information System (INIS)

Fibroblast cells derived from a cafe-au-lait spot and normal-appearing skin of a neurofibromatosis (NF-6) patient were studied for radiosensitivity in comparison with two normal cell lines used as controls. No difference in radiosensitivity was observed between the patient's cell lines and the controls using acute gamma-irradiation. However, a markedly increased radiosensitivity of the fibroblasts obtained from the patient's skin of normal appearance was demonstrated after chronic gamma-irradiation. The cells from the cafe-au-lait spot showed intermediate sensitivity to chronic irradiation as compared with the control cell lines and the fibroblasts derived from the normal skin of the patient. These results showed the usefulness of chronic irradiation in detecting increased cellular radiosensitivity which may result from a unique DNA repair defect in an NF patient. We suggest that enhanced genetic changes in radiosensitive NF patients may lead to formation of cafe-au-lait lesions and certain tumors. Such a transformation may be associated with production of radiotolerant cells

1990-07-15

66

Radiosensitivity of fibroblasts obtained from a cafe-au-lait spot and normal-appearing skin of a patient with neurofibromatosis (NF-6)  

Energy Technology Data Exchange (ETDEWEB)

Fibroblast cells derived from a cafe-au-lait spot and normal-appearing skin of a neurofibromatosis (NF-6) patient were studied for radiosensitivity in comparison with two normal cell lines used as controls. No difference in radiosensitivity was observed between the patient's cell lines and the controls using acute gamma-irradiation. However, a markedly increased radiosensitivity of the fibroblasts obtained from the patient's skin of normal appearance was demonstrated after chronic gamma-irradiation. The cells from the cafe-au-lait spot showed intermediate sensitivity to chronic irradiation as compared with the control cell lines and the fibroblasts derived from the normal skin of the patient. These results showed the usefulness of chronic irradiation in detecting increased cellular radiosensitivity which may result from a unique DNA repair defect in an NF patient. We suggest that enhanced genetic changes in radiosensitive NF patients may lead to formation of cafe-au-lait lesions and certain tumors. Such a transformation may be associated with production of radiotolerant cells.

Hannan, M.A.; Smith, B.P.; Sigut, D.; Sackey, K. (King Faisal Specialist Hospital and Research Centre, Riyadh (Saudi Arabia))

1990-07-15

67

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)"2D"3 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)"2D"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 [_3H]1,25(OH)"2D"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)"2D resulted from five or six distinct genetic mutations in six kindreds

1983-01-01

68

Human skin keratinocytes, melanocytes, and fibroblasts contain distinct circadian clock machineries.  

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Skin acts as a barrier between the environment and internal organs and performs functions that are critical for the preservation of body homeostasis. In mammals, a complex network of circadian clocks and oscillators adapts physiology and behavior to environmental changes by generating circadian rhythms. These rhythms are induced in the central pacemaker and peripheral tissues by similar transcriptional-translational feedback loops involving clock genes. In this work, we investigated the presence of functional oscillators in the human skin by studying kinetics of clock gene expression in epidermal and dermal cells originating from the same donor and compared their characteristics. Primary cultures of fibroblasts, keratinocytes, and melanocytes were established from an abdominal biopsy and expression of clock genes following dexamethasone synchronization was assessed by qPCR. An original mathematical method was developed to analyze simultaneously up to nine clock genes. By fitting the oscillations to a common period, the phase relationships of the genes could be determined accurately. We thereby show the presence of functional circadian machinery in each cell type. These clockworks display specific periods and phase relationships between clock genes, suggesting regulatory mechanisms that are particular to each cell type. Taken together, our data demonstrate that skin has a complex circadian organization. Oscillators are present not only in fibroblasts but also in epidermal keratinocytes and melanocytes and are likely to act in coordination to drive rhythmic functions within the skin. PMID:22627494

Sandu, Cristina; Dumas, Marc; Malan, André; Sambakhe, Diariétou; Marteau, Clarisse; Nizard, Carine; Schnebert, Sylvianne; Perrier, Eric; Challet, Etienne; Pévet, Paul; Felder-Schmittbuhl, Marie-Paule

2012-10-01

69

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

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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 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.)

1998-06-01

70

Preventive effects of tamarind seed coat extract on UVA-induced alterations in human skin fibroblasts.  

Science.gov (United States)

One of the most damaging actions on skin is from solar radiation, particularly from its ultraviolet (UV) component, through the formation of oxidative species. Thus, an antioxidant strategy that prevents the formation of these oxidants could form the basis of an efficacious cutaneous protectant. Many herbal materials contain antioxidant polyphenols, and this study assessed the possibility that tamarind seed coat extract could fulfill this role. An alcoholic extract of the tamarind (Tamarindus indica L.) seed coat showed stronger antioxidant activity (2,2-diphenyl-1-picrylhydrazyl inhibition, EC(50) = 12.9 ?g/ml) than L-ascorbic acid (EC(50) = 22.9 ?g/ml) and ?-tocopherol (EC(50) = 29.3 ?g/ml). In cultured fibroblasts taken from human skin, hydrogen peroxide (100-1000 ?M) damaged 62-92% of the cells compared to only 35-47% when the cells were preincubated in extract (200 ?g/ml) for 24 h. UVA (40 J/cm2) irradiation of human fibroblasts damaged 25% of the cells but the death rate was reduced to 10% with extract. UV irradiation increased the proportion of cells arrest in G(0)/G(1) phase (from 59% to 78%) but this was largely prevented by the extract (64%), according to flow cytometry. Intracellular total glutathione of UVA-irradiated cells pretreated with the extract increased to 10-25% compared to the non-pretreated group at 24-72 h after irradiation. Fibroblasts typically increased matrix metalloproteinase-1 secretion after photodamage, and this is prevented by the extract. This is the first report showing that tamarind seed coat extract is an antioxidant and can protect human skin fibroblasts from cellular damage produced by UVA and thus may form the foundation for an antiaging cosmetic. PMID:24602819

Phetdee, Khemjira; Rakchai, Racharat; Rattanamanee, Kwanchai; Teaktong, Thanasak; Viyoch, Jarupa

2014-01-01

71

[Development of an engraftable skin equivalent based on matriderm with human keratinocytes and fibroblasts].  

Science.gov (United States)

A cell-based wound coverage with keratinocytes and fibroblasts on the basis of a commercially available dermal substitute (Matriderm ((R)), Kollagen/Elastin matrix) was generated, in order to treat wide burn wounds. First the expansion of keratinocytes was optimised and the culturing time was minimised. Raw material was 1-2 cm (2) split skin. Dermis and epidermis were separated by enzymatic treatment with thermolysin. After treatment of both compartments with trypsin and collagenase I, keratinocytes and fibroblasts were isolated and expanded in collagen I coated dishes. After 10 days fibroblasts were seeded on Matriderm ((R)). After cultivation of the fibroblasts-containing matrix for one week keratinocytes were seeded on top. After an additional week of submersed cultivation the matrix was lifted up to the air-liquid interface to initiate epidermal cell differentiation. After 16 days in the air-liquid interphase the matrix was fixed and underwent immunohistochemical and electron microscopic analysis. Histological analysis showed a regularly stratification of the epidermal part. We observed collagen IV, a marker for the basement membrane, between epidermis and dermis. Desmoglein and the differentiation markers involucrine and cytokeratin 10 were found in the suprabasal layers of the epidermis. Electron microscopic analysis showed the basement membrane in the epidermal junction zone as well as cell-cell connections in the form of desmosomes. Late differentiation characteristics, like granular structures and the cornified layer, were found in the stratum granulosum and stratum corneum. Our results demonstrate that a skin equivalent can be generated by using a collagen/elastin matrix, with an expansion rate of 50-100-fold. This skin equivalent may be useful for covering deep wounds. PMID:19711256

Golinski, P A; Zöller, N; Kippenberger, S; Menke, H; Bereiter-Hahn, J; Bernd, A

2009-12-01

72

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

73

Hexavalent chromium-induced alteration of proteomic landscape in human skin fibroblast cells.  

Science.gov (United States)

Hexavalent chromium [Cr(VI)] generated during industrial processes is carcinogenic. Although much is known about the deleterious effects caused by reactive oxygen species generated during the reduction of Cr(VI) after its absorption by biological systems, the precise mechanisms underlying Cr(VI) cytotoxicity remain poorly defined. Here, we analyzed, at the global proteome scale, the perturbation of protein expression in GM00637 human skin fibroblast cells upon exposure to potassium dichromate (K?Cr?O?). We were able to quantify ?4600 unique proteins, among which ?400 exhibited significant alterations in expression levels upon a 24-h treatment with 0.5 ?M K?Cr?O?. Pathway analysis revealed the Cr(VI)-induced perturbation of cholesterol biosynthesis, G-protein signaling, inflammatory response, and selenoprotein pathways. In particular, we discovered that the K?Cr?O? treatment led to pronouncedly elevated expression of a large number of enzymes involved in de novo cholesterol biosynthesis. Real-time PCR analysis revealed the increased mRNA expression of selected genes involved in cholesterol biosynthesis. Consistently, K?Cr?O? treatment resulted in marked increases in cellular cholesterol level in multiple cell lines. Moreover, the Cr(VI)-induced growth inhibition of cultured human cells could be rescued by a cholesterol-lowering drug, lovastatin. Together, we demonstrated, for the first time, that Cr(VI) may exert its cytotoxic effect, at least partly, through the up-regulation of enzymes involved in de novo cholesterol biosynthesis and the resultant increase of cholesterol level in cells. PMID:23718831

Guo, Lei; Xiao, Yongsheng; Wang, Yinsheng

2013-07-01

74

Artificial Skin - Culturing of Different Skin Cell Lines for Generating an Artificial Skin Substitute on Cross-Weaved Spider Silk Fibres  

Science.gov (United States)

Background In the field of Plastic Reconstructive Surgery the development of new innovative matrices for skin repair is in urgent need. The ideal biomaterial should promote attachment, proliferation and growth of cells. Additionally, it should degrade in an appropriate time period without releasing harmful substances, but not exert a pathological immune response. Spider dragline silk from Nephila spp meets these demands to a large extent. Methodology/Principal Findings Native spider dragline silk, harvested directly out of Nephila spp spiders, was woven on steel frames. Constructs were sterilized and seeded with fibroblasts. After two weeks of cultivating single fibroblasts, keratinocytes were added to generate a bilayered skin model, consisting of dermis and epidermis equivalents. For the next three weeks, constructs in co-culture were lifted on an originally designed setup for air/liquid interface cultivation. After the culturing period, constructs were embedded in paraffin with an especially developed program for spidersilk to avoid supercontraction. Paraffin cross- sections were stained in Haematoxylin & Eosin (H&E) for microscopic analyses. Conclusion/Significance Native spider dragline silk woven on steel frames provides a suitable matrix for 3 dimensional skin cell culturing. Both fibroblasts and keratinocytes cell lines adhere to the spider silk fibres and proliferate. Guided by the spider silk fibres, they sprout into the meshes and reach confluence in at most one week. A well-balanced, bilayered cocultivation in two continuously separated strata can be achieved by serum reduction, changing the medium conditions and the cultivation period at the air/liquid interphase. Therefore spider silk appears to be a promising biomaterial for the enhancement of skin regeneration.

Reimers, Kerstin; Kuhbier, Joern W.; Schafer-Nolte, Franziska; Allmeling, Christina; Kasper, Cornelia; Vogt, Peter M.

2011-01-01

75

Ultrastructural changes of mitochondria in the cultivated skin fibroblasts of patients with point mutations in mitochondrial DNA.  

Science.gov (United States)

Mitochondrial disorders represent a heterogeneous group of multisystem diseases with extreme variability in clinical phenotype. The diagnosis of mitochondrial disorders relies heavily on extensive biochemical and molecular analyses combined with morphological studies including electron microscopy. Although muscle is the tissue of choice for electron microscopic studies, the authors investigated cultivated human skin fibroblasts (HSF) harboring 3 different pathologic mtDNA mutations: 3243A > G, 8344A > G, 8993T > G. They addressed to the possibility of whether mtDNA mutations influence mitochondrial morphology in HSF and if ultrastructural changes of mitochondria may be used for differential diagnostics of mitochondrial disorders caused by mtDNA mutations. Ultrastructural analysis of patients' HSF revealed a heterogeneous mixture of mainly abnormal, partially swelling mitochondria with unusual and sparse cristae. The most characteristic cristal abnormalities were heterogeneity in size and shapes or their absence. Typical filamentous and branched mitochondria with numerous cristae as appeared in control HSF were almost not observed. In all lines of cultured HSF with various mtDNA mutations, similar ultrastructural abnormalities and severely changed mitochondrial interior were found, although no alterations in function and amount of OXPHOS were detected by routinely used biochemical methods in two lines of cultured HSF. This highlights the importance of morphological analysis, even in cultured fibroblasts, in diagnostics of mitochondrial disorders. PMID:16971348

Brantová, Olga; Tesarová, Markéta; Hansíková, Hana; Elleder, Milan; Zeman, Jirí; Sládková, Jana

2006-01-01

76

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

77

Use of a genetic variant to study the hexose transport properties of human skin fibroblasts.  

Science.gov (United States)

Human skin fibroblasts from 'normal' subjects were found to possess at least two hexose transport systems. One system was responsible for the uptake of 2-deoxy-D-glucose (dGlc), D-glucose and D-galactose, whereas the other was responsible primarily for the uptake of 3-O-methyl-D-glucose (MeGlc). The transport of dGlc was the rate-limiting step in the uptake process; over 97% of the internalized dGlc was phosphorylated and the specific activity of hexokinase was several times higher than that for dGlc transport. The dGlc transport system was activated by glucose starvation, and was very sensitive to inhibition by cytochalasin B and energy uncouplers. Fibroblasts isolated from a patient with symptoms of hypoglycaemia were found to differ from their normal counterparts in the dGlc transport system. They exhibited a much higher transport affinity for dGlc, D-glucose and D-galactose, with no change in the respective transport capacity. Transport was not the rate-limiting step in dGlc uptake by these cells. Moreover, the patient's dGlc transport system was no longer sensitive to inhibition by cytochalasin B and energy uncouplers. This suggested that the intrinsic properties of the patient's dGlc transport system were altered. It should be noted that the patient's dGlc transport system could still be activated by glucose starvation. Despite the changes in the dGlc transport system, the MeGlc transport system in the patient's fibroblasts remained unaltered. The observed difference in the properties of the two hexose transport systems in the 'normal' and the patient's fibroblasts strongly suggests that the two transport systems may be coded or regulated by different genes. The present finding provides the first genetic evidence from naturally occurring fibroblasts indicating the presence of two different hexose transport systems. PMID:2306216

Mesmer, O T; Gordon, B A; Rupar, C A; Lo, T C

1990-02-01

78

The fibroblast-derived paracrine factor neuregulin-1 has a novel role in regulating the constitutive color and melanocyte function in human skin.  

Science.gov (United States)

Interactions between melanocytes and neighboring cells in the skin are important in regulating skin color in humans. We recently demonstrated that the less pigmented and thicker skin on the palms and soles is regulated by underlying fibroblasts in those areas, specifically via a secreted factor (DKK1) that modulates Wnt signaling. In this study, we tested the hypothesis that dermal fibroblasts regulate the constitutive skin color of individuals ranging from very light to very dark. We used microarray analysis to compare gene expression patterns in fibroblasts derived from lighter skin types compared to darker skin types, with a focus on secreted proteins. We identified a number of genes that differ dramatically in expression and, among the expressed proteins, neuregulin-1, which is secreted by fibroblasts derived from dark skin, effectively increases the pigmentation of melanocytes in tissue culture and in an artificial skin model and regulates their growth, suggesting that it is one of the major factors determining human skin color. PMID:20736300

Choi, Wonseon; Wolber, Rainer; Gerwat, Wolfram; Mann, Tobias; Batzer, Jan; Smuda, Christoph; Liu, Hongfang; Kolbe, Ludger; Hearing, Vincent J

2010-09-15

79

Accumulation of phosphorus compounds in tissues and cultured skin fibroblasts in patients with hypophosphatasia  

International Nuclear Information System (INIS)

Patients with hypophosphatasia caused by a deficiency of alkaline phosphatase first showed marked accumulation of phosphoethanolamine and other phosphorus compounds in kidney and liver, while in placenta and intestine contents of these compounds were within a normal range. Furthermore, 32P-incorporation in cultured skin fibroblasts of patients with hypophosphatasia was increased about two to three times of control. FPLC chromatographic analysis also indicates that the accumulated phosphorus compounds in hypophosphatasia was smaller molecular phosphorus containing compounds. These data provide new pathophysiological aspect of hypophosphatasia

1988-08-15

80

Membrane damage induced in cultured human skin fibroblasts by UVA irradiation  

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Irradiation of cultured human skin fibroblasts with ultraviolet light from 320 to 400 nm (UVA) leads to a decrease in the membrane fluidity exemplified by an enhanced fluorescence anisotropy of the lipophilic fluorescent probe 1-[4-(trimethylamino)-phenyl]-6-phenylhexa-1,3,5-triene. This UVA-induced decrease in fluidity is associated with lactate dehydrogenase leakage in the supernatant. Vitamin E, an inhibitor of lipid peroxidation, exerts a protective effect on both phenomena. Therefore, this UVA-induced damage in membrane properties may be related to lipid peroxidation processes. Moreover, exponentially growing cells are more sensitive to these UVA-induced alterations than confluent cells. (Author).

Gaboriau, F. (Institut Curie, 75 - Paris (France) Paris-6 Univ., 75 (France) Hopital Henri-Mondor, 94 - Creteil (France)); Morliere, P.; Marquis, I.; Moysan, A. (Hopital Henri-Mondor, 94 - Creteil (France)); Geze, M. (Museum National d' Histoire Naturelle, 75 - Paris (France)); Dubertret, L. (Hopital Henri-Mondor, 94 - Creteil (France) Hopital Saint-Louis, 75 - Paris (France))

1993-10-01

 
 
 
 
81

Human skin fibroblast stromelysin: structure, glycosylation, substrate specificity, and differential expression in normal and tumorigenic cells  

International Nuclear Information System (INIS)

The authors have purified and determined the complete primary structure of human stromelysin, a secreted metalloprotease with a wide range of substrate specificities. Human stromelysin is synthesized in a preproenzyme form with a calculated size of 53,977 Da and a 17-amino acid long signal peptide. Prostromelysin is secreted in two forms, with apparent molecular masses on NaDodSO4/PAGE of 60 and 57 kDa. Human stromelysin is capable of degrading proteoglycan, fibronectin, laminin, and type IV collagen but not interstitial type I collagen. The enzyme is not capable of activating purified human fibroblast procollagenase. Analysis of its primary structure shows that stromelysin is in all likelihood the human analog of rat transin, which is an oncogene transformation-induced protease. The pattern of enzyme expression in normal and tumorigenic cells revealed that human skin fibroblasts in vitro secrete stromelysin constitutively. Human fetal lung fibroblasts transformed with simian virus 40, human bronchial epithelial cells transformed with the ras oncogene, fibrosarcoma cells (HT-1080), and a melanoma cell strain (A 2058), do not express this protease nor can the enzyme be induced in these cells by treatment with phorbol 12-myristate 13-acetate. The data indicate that the expression and the possible involvement of secreted metalloproteases in tumorigenesis result from a specific interaction between the transforming factor and the target cell, which may vary in different species

1987-01-01

82

Nerve growth factor displays stimulatory effects on human skin and lung fibroblasts, demonstrating a direct role for this factor in tissue repair  

Science.gov (United States)

Nerve growth factor (NGF) is a polypeptide which, in addition to its effect on nerve cells, is believed to play a role in inflammatory responses and in tissue repair. Because fibroblasts represent the main target and effector cells in these processes, to investigate whether NGF is involved in lung and skin tissue repair, we studied the effect of NGF on fibroblast migration, proliferation, collagen metabolism, modulation into myofibroblasts, and contraction of collagen gel. Both skin and lung fibroblasts were found to produce NGF and to express tyrosine kinase receptor (trkA) under basal conditions, whereas the low-affinity p75 receptor was expressed only after prolonged NGF exposure. NGF significantly induced skin and lung fibroblast migration in an in vitro model of wounded fibroblast and skin migration in Boyden chambers. Nevertheless NGF did not influence either skin or lung fibroblast proliferation, collagen production, or metalloproteinase production or activation. In contrast, culture of both lung and skin fibroblasts with NGF modulated their phenotype into myofibroblasts. Moreover, addition of NGF to both fibroblast types embedded in collagen gel increased their contraction. Fibrotic human lung or skin tissues displayed immunoreactivity for NGF, trkA, and p75. These data show a direct pro-fibrogenic effect of NGF on skin and lung fibroblasts and therefore indicate a role for NGF in tissue repair and fibrosis.

Micera, Alessandra; Vigneti, Eliana; Pickholtz, Dalia; Reich, Reuven; Pappo, Orit; Bonini, Sergio; Maquart, François Xavier; Aloe, Luigi; Levi-Schaffer, Francesca

2001-05-01

83

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

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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

84

Action spectra for inactivation of normal and xeroderma pigmentosum human skin fibroblasts by ultraviolet radiations  

International Nuclear Information System (INIS)

Action spectra for UV-induced lethality as measured by colony forming ability were determined both for a normal human skin fibroblast strain (1BR) and for an excision deficient xeroderma pigmentosum strain (XP4LO) assigned to complementation group A using 7 monochromatic wavelengths in the range 254-365 nm. The relative sensitivity of the XP strain compared to the normal skin fibroblasts shows a marked decrease at wavelengths longer than 313 nm, changing from a ratio of about 20 at the shorter wavelengths to just greater than 1.0 at the longer wavelengths. The action spectra thus indicate that the influence on cell inactivation of the DNA repair defect associated with XP cells is decreased and almost reaches zero at longer UV wavelengths. This would occur, for example, if the importance of pyrimidine dimers as the lethal lesion decreased with increasing wavelength. These results are consistent with pyrimidine dimers induced in DNA being the major lethal lesion in both cell strains over the wavelength range 254-313 nm. However, it is indicated that different mechanisms of inactivation operate at wavelengths longer than 313 nm. (author)

1983-01-01

85

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

86

Sensitivity of cultured skin fibroblasts from patients with neurofibromatosis to DNA-damaging agents  

International Nuclear Information System (INIS)

Neurofibromatosis (NF) is an autosomal dominant disorder associated with various constitutional abnormalities as well as a striking predisposition for malignant and nonmalignant neoplasms, both in cells originating in and not originating in the neural crest. We have examined the sensitivity of cultured skin fibroblasts from patients with neurofibromatosis to several types of DNA damage. Fibroblasts in Dulbecco's modified Eagle's medium were plated at 10(2) to 2 X 10(4) cells per 75 cm2 tissue culture plates, and exposed to various doses of gamma radiation (leads to DNA scission), actinomycin D, or mitomycin C. Cells were reincubated for 15 to 40 days until surviving colonies exhibited greater than 30-50 cells. Plates were then stained with 1% methylene blue and the colonies counted, with surviving fraction determined relative to plating efficiency. Nine skin fibroblast cell strains from normal individuals were studied as controls. One neurofibromatosis (NF) cell strain, SB23, exhibited normal sensitivity to all three DNA-damaging agents studied in early (7-8) and middle (12-13) in vitro passage. Strain GM0622, on the other hand, exhibited normal sensitivity to the three DNA-damaging agents studied at early passage, but showed a significant decrease in survival after exposure to both gamma radiation (D0 = 106 rad) and actinomycin D (D0 = 0.024 mcg/ml) with increasing passage. Strain GM1639 exhibited decreased survival after actinomycin D exposure at early passage (D0 = 0.017 mcg/ml), with normal survival after exposure to gamma radiation and mitomycin C at the same passage

1986-01-01

87

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 "1"4C-protein hydrolysate, ("1"4C)uridine, and ("1"4C) thymidine. Stimulation was determined by measuring incorporation of ("1"4C)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

1985-07-01

88

The fibroblast-derived paracrine factor neuregulin-1 has a novel role in regulating the constitutive color and melanocyte function in human skin  

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Interactions between melanocytes and neighboring cells in the skin are important in regulating skin color in humans. We recently demonstrated that the less pigmented and thicker skin on the palms and soles is regulated by underlying fibroblasts in those areas, specifically via a secreted factor (DKK1) that modulates Wnt signaling. In this study, we tested the hypothesis that dermal fibroblasts regulate the constitutive skin color of individuals ranging from very light to very dark. We used mi...

Choi, Wonseon; Wolber, Rainer; Gerwat, Wolfram; Mann, Tobias; Batzer, Jan; Smuda, Christoph; Liu, Hongfang; Kolbe, Ludger; Hearing, Vincent J.

2010-01-01

89

Keratinocytes contract human dermal extracellular matrix and reduce soluble fibronectin production by fibroblasts in a skin composite model.  

Science.gov (United States)

Composites of human de-epidermised acellular dermis and normal adult human keratinocytes and fibroblasts were examined for the ability of cells to contract these composites. Image analysis of the outline of the composites showed that, in this model, keratinocytes alone or in the presence of fibroblasts caused highly significant contraction (of the order of 25% by day 12). There was no significant contraction of the dermis with fibroblasts alone or in the absence of cells. The presence or absence of basement membrane antigens did not influence the effect of keratinocytes on dermal contraction. Analysis of the conditioned media from these composites showed that the greatest fibronectin production was seen with fibroblasts alone in the presence of basement membrane. Keratinocytes alone produced little fibronectin irrespective of the presence of the basement membrane. If keratinocytes were present with fibroblasts, however, then fibronectin production was significantly reduced both in the presence and absence of the basement membrane, indicating that keratinocytes modify dermal fibroblast extracellular matrix production. This study shows that while keratinocytes and fibroblasts are clearly influencing each other's activity in this human skin composite model, under the circumstances we describe it is the keratinocyte and not the fibroblast which causes contraction of the human de-epidermised acellular dermis. PMID:9326143

Ralston, D R; Layton, C; Dalley, A J; Boyce, S G; Freedlander, E; MacNeil, S

1997-09-01

90

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

Directory of Open Access Journals (Sweden)

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

91

Differential Regulation of a Fibroblast Growth Factor-Binding Protein during Skin Carcinogenesis and Wound Healing1  

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The initiation of premalignant lesions is associated with subtle cellular and gene expression changes. Here we describe a severe combined immunodeficient mouse xenograft model with human adult skin and compare chemical carcinogenesis and wound healing. We focus on a secreted binding protein for fibroblast growth factors (FGF-BP) that enhances the activity of locally stored FGFs and is expressed at high levels in human epithelial cancers. Carcinogen treatment of murine skin induced papilloma w...

Kurtz, Andreas; Aigner, Achim; Cabal-manzano, Rafael H.; Butler, Robert E.; Hood, Dozier R.; Sessions, Roy B.; Czubayko, Frank; Wellstein, Anton

2004-01-01

92

Effect of growth factors on dermal fibroblast contraction in normal skin and hypertrophic scar.  

Science.gov (United States)

We have examined the effects of four 'exogenous' growth factors, i.e. PDGF-BB (5 ng/ml), TGF-beta1 (5 ng/ml), bFGF (10 ng/ml) and EGF (10 ng/ml) on the contraction of floating collagen type I lattices populated by human normal skin (NS) and hypertrophic scar (HS) fibroblasts (FPCL). Only TGF-beta1 enhanced the contractility of both NS and HS fibroblasts in the collagen lattice (P 0.05). The onset effect of TGF-beta1 on NS-FPCL contraction was relative early at 24 h after FPCL casting as compared to a 72 h delay on HS-FPCL contraction. Besides, PDGF-BB was found to be able to enhance HS-FPCL contraction (P FPCL contraction on day 4. On the other hand, three enzyme-linked immunosorbent assays (ELISA) were performed to demonstrate quantitatively the 'endogenous' growth factors that fibroblasts secreted into the culture medium 48 h after FPCL casting. No appreciable difference was found between 10 NS and 11 HS samples tested for PDGF-AB immunoassay (11.48 +/- 5.5 pg/ml versus 12.20 +/- 5.34 pg/ml). The same result existed in 7 NS and 13 HS samples for TGF-beta2 immunoassay (15.15 +/- 6.2 pg/ml versus 11.84 +/- 7.46 pg/ml). In bFGF immunoassay study, relative variable data was noted in both 7 NS (18.18 +/- 13.18 pg/ml) and 12 HS samples (20.41 +/- 22.36 pg/ml). In conclusion, we suppose that TGF-beta role in wound healing may be due to the secondary exogenous influences. The endogenous ability of TGF-beta2 secretion (quantity) in HS fibroblasts are the same as NS fibroblasts but with delayed timing responses (quality) to exogenous TGF-beta1 effect in the collagen lattice. Further studies with timing-regulated selective specific monoclonal antibodies against the growth factor receptors may provide the therapeutic applications on HS during wound healing. PMID:9039980

Yang, C C; Lin, S D; Yu, H S

1997-02-01

93

Cerebrohepatorenal Syndrome (CHRS) of Zellweger: lysosomal enzyme activities, sulfation of glycosaminoglycans, and pipecolic acid levels in cultured skin fibroblasts  

International Nuclear Information System (INIS)

The defect in the cerebrohepatorenal syndrome (CHRS), a fatal hereditary disorder primarily affecting neurological development, is unknown. Three areas were studied for specific biochemical abnormalities which might aid in diagnosis and understanding of the disorder: (1) Clinico-pathological similarities to inherited degenerative neurologic disorders suggested decreased activity of certain lysosomal enzymes. Assays of ?-galactosidase, ?-hexosaminidase, ?-mannosidase, and arylsulfatase A activities in fibroblasts from four infants with CHRS indicated no deficiency of enzyme activities. (2) Undersulfation of glycosaminoglycans (GAGs) has been reported in patients with the clinically similar Lowe's syndrome. The rate and amount of incorporation of "3"5SO_4 = into intracellular "3"5S-GAGs up to 48 hours was comparable in fibroblasts from six CHRS infants and controls. Loss of "3"5-GAGs also followed a normal pattern. (3) Because pipecolic acid (PA) has been reported to be elevated in body fluids of patients with CHRS, cultured skin fibroblasts were examined for such an abnormality. Lysosomal enzyme activities and metabolism of sulfated glycosaminoglycans appear to be normal in cultured skin fibroblasts from infants with CHRS. Despite the sensitivity of the method, examination of pipecolic acid in cultured skin fibroblasts does not seem to be useful for diagnosis of CHRS

1985-01-01

94

Immortalized human skin fibroblast feeder cells support growth and maintenance of both human embryonic and induced pluripotent stem cells  

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BACKGROUND: Feeder cells are frequently used for the early-stage of derivation and culture of human embryonic stem cell (hESC) lines. METHODS: We established a conditionally immortalized human foreskin fibroblast line that secreted basic fibroblast growth factor (bFGF). These cells were used as feeder cells for hESC culture and induced pluripotent stem (iPS) cell derivation and expansion. This conditional immortalization was performed using lentiviral vector (LV) mediated transduction of Bmi-...

Unger, C.; Gao, S.; Cohen, M.; Jaconi, M.; Bergstrom, R.; Holm, F.; Galan, A.; Sanchez, E.; Irion, O.; Dubuisson, J. B.; Giry-laterriere, M.; Salmon, P.; Simon, C.; Hovatta, O.; Feki, A.

2009-01-01

95

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.)

1982-01-01

96

Regulation and inhibition of collagenase expression by long-wavelength ultraviolet radiation in cultured human skin fibroblasts  

International Nuclear Information System (INIS)

The cellular mechanisms responsible for the connective tissue changes produced by chronic exposure to UV light are poorly understood. collagenase, a metalloproteinase, initiates degradation of types I and III collagen and thus plays a key role in the remodeling of dermal collagen. Collagenase synthesis by fibroblasts and keratinocytes involves the protein kinase C (PKC) second messenger system, and corticosteroids have been shown to suppress its synthesis at the level of gene transcription. Long-wavelength UV light (UVA, 320-400 nm) stimulates the synthesis of interstitial collagenase, as well as increasing PKC activity, in human skin fibroblasts in vitro. This study explores the regulation of collagenase expression by UVA in cultured human skin fibroblasts. Specifically, the time course, the effect of actinomycin D, an inhibitor of RNA synthesis, as well as the effect of PKC inhibitors and dexamethansone on expression of collagenase following UVA irradiation were examined. (Author)

1995-09-01

97

Regulation and inhibition of collagenase expression by long-wavelength ultraviolet radiation in cultured human skin fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

The cellular mechanisms responsible for the connective tissue changes produced by chronic exposure to UV light are poorly understood. collagenase, a metalloproteinase, initiates degradation of types I and III collagen and thus plays a key role in the remodeling of dermal collagen. Collagenase synthesis by fibroblasts and keratinocytes involves the protein kinase C (PKC) second messenger system, and corticosteroids have been shown to suppress its synthesis at the level of gene transcription. Long-wavelength UV light (UVA, 320-400 nm) stimulates the synthesis of interstitial collagenase, as well as increasing PKC activity, in human skin fibroblasts in vitro. This study explores the regulation of collagenase expression by UVA in cultured human skin fibroblasts. Specifically, the time course, the effect of actinomycin D, an inhibitor of RNA synthesis, as well as the effect of PKC inhibitors and dexamethansone on expression of collagenase following UVA irradiation were examined. (Author).

Petersen, Marta; Hamilton, Tiffani; Haili Li [Utah Univ., Salt Lake City, UT (United States). Dept. of Internal Medicine

1995-09-01

98

Quantitative assessment of changes in the dermal fibroblast population of pig skin after single doses of X-rays  

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Changes in the density of fibroblast nuclei in reticular dermis of pigs was studied from 6 to 104 weeks after a single dose of 15.4 Gy of X-rays. The largest decrease in fibroblasts occurred between 12 and 26 weeks after irradiation; after this there was only a slight fall in fibroblast number until 104 weeks when observations ceased. At 26 weeks and later times after irradiation reduction in the density of fibroblast nuclei in the reticular dermis was dose-dependent for single doses in the range 8.0-20.7 Gy. The dose-response curve had an initial shoulder, after which the fall in the fibroblast nuclear density was linearly related to dose. Data obtained between 26 weeks and 104 weeks after irradiation, could be fitted by the same dose-response curve. The fall in the counts of fibroblast nuclei was compared with earlier studies. The loss of fibroblasts occurred after an initial reduction in blood flow in the pig skin but was concomitant with general reduction in dermal thickness.

Hamlet, R.; Hopewell, J.W.

1988-10-01

99

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

100

The Protective Effects of Fucosterol Against Skin Damage in UVB-Irradiated Human Dermal Fibroblasts.  

Science.gov (United States)

Exposure to ultraviolet (UV) light causes matrix metalloproteinase (MMP) overexpression and extracellular matrix depletion, leading to skin photoaging. The activation of MMP is related to increased interlukin-6 (IL-6) and type I procollagen production, which is regulated by transforming growth factor-?1 (TGF-?1). Activator protein-1 (AP-1) activation induces MMP-1 production and reduces type I procollagen secretion. Fucosterol, which is extracted and purified from the brown algae Hizikia fusiformis, is a phytosterol. We assessed the effects of fucosterol on photodamage and investigated its molecular mechanism of action in UVB-irradiated normal human dermal fibroblasts by using enzyme-linked immunosorbent assay, Western blot analysis, and reverse transcription-polymerase chain reaction. Our results showed that fucosterol significantly decreased the UVB-induced expression of MMP-1, IL-6, p-c-Jun, and p-c-Fos. Additionally, fucosterol markedly increased the UVB-induced production of type I procollagen and TGF-?1. Our results indicate that fucosterol regulates MMP-1 and type I procollagen expression by modulating AP-1 and TGF-?1 signaling and that MMP-1 activation is correlated with IL-6. These data suggest that fucosterol is a promising botanical agent to protect against skin photodamage. PMID:24142195

Hwang, Eunson; Park, Sang-Yong; Sun, Zheng-Wang; Shin, Heon-Sub; Lee, Don-Gil; Yi, Tae Hoo

2014-06-01

 
 
 
 
101

Skin fibroblasts as a tool for identifying the risk of nephropathy in the type 1 diabetic population.  

Science.gov (United States)

Human fibroblasts in culture have been employed as an in vitro system to investigate some pathophysiological mechanisms of diabetes mellitus also associated with the development of diabetic nephropathy. In fact, there is increasing evidence that genetic factors either convey the risk of, or protect from, diabetic nephropathy and that the expression profiles and/or the behaviour of the cultured skin fibroblasts from type 1 diabetic patients could reflect these genetic influences. On the other hand, alterations could be attributable not only to changes in DNA sequence, but also to epigenetic factors. Our aim is to make a critical overview of the studies involving primary cultures of skin fibroblasts as tools to investigate the pathophysiology of diabetic nephropathy performed until now in this area. Cultured skin fibroblasts could be useful not only for the identification of patients at risk of developing diabetic renal disease, but also for a better understanding of the complex multifactorial mechanisms leading to the long-term complications in diabetes. PMID:22218755

Millioni, Renato; Puricelli, Lucia; Iori, Elisabetta; Trevisan, Roberto; Tessari, Paolo

2012-01-01

102

Normal rejoining of DNA strand breaks in ataxia telangiectasia fibroblast lines after low x-ray exposure  

International Nuclear Information System (INIS)

The alkaline elution method was used to measure the enzymatic repair of x-ray-induced DNA strand breaks in skin fibroblasts derived from human subjects afflicted with ataxia telangiectasia (AT). Monolayer cultures of normal control and AT cell lines were exposed acutely to moderately lethal (250-rad) and highly lethal (1250-rad) doses of 250-kV x rays under aerobic conditions. Upon receiving 250 rad, the control fibroblasts from a clinically normal donor rejoined all detectable single-strand breaks (plus alkali-labile bonds) within 30 to 60 min of incubation. When challenged with 1250 rad the kinetics of strand rejoining by the normal control cells were biphasic. For both exposures, no significant difference in either the rate or the extent of strand rejoining was detected between the normal cell line (GM38) and three mutant cell lines (AT2BE, AT3BI, AT4BI) belonging to the three known genetic complementation groups in AT. It would thus appear that the enhanced radiosensitivity of cultured AT cells does not stem from faulty rejoining of radiogenic DNA strand breaks

1981-01-01

103

Full-thickness skin wound healing using autologous keratinocytes and dermal fibroblasts with fibrin: bilayered versus single-layered substitute.  

Science.gov (United States)

Split-skin grafting (SSG) is the gold standard treatment for full-thickness skin defects. For certain patients, however, an extensive skin lesion resulted in inadequacies of the donor site. Tissue engineering offers an alternative approach by using a very small portion of an individual's skin to harvest cells for propagation and biomaterials to support the cells for implantation. The objective of this study was to determine the effectiveness of autologous bilayered tissue-engineered skin (BTES) and single-layer tissue-engineered skin composed of only keratinocytes (SLTES-K) or fibroblasts (SLTES-F) as alternatives for full-thickness wound healing in a sheep model. Full-thickness skin biopsies were harvested from adult sheep. Isolated fibroblasts were cultured using medium Ham's F12: Dulbecco modified Eagle medium supplemented with 10% fetal bovine serum, whereas the keratinocytes were cultured using Define Keratinocytes Serum Free Medium. The BTES, SLTES-K, and SLTES-F were constructed using autologous fibrin as a biomaterial. Eight full-thickness wounds were created on the dorsum of the body of the sheep. On 4 wounds, polyvinyl chloride rings were used as chambers to prevent cell migration at the edge. The wounds were observed at days 7, 14, and 21. After 3 weeks of implantation, the sheep were euthanized and the skins were harvested. The excised tissues were fixed in formalin for histological examination via hematoxylin-eosin, Masson trichrome, and elastin van Gieson staining. The results showed that BTES, SLTES-K, and SLTES-F promote wound healing in nonchambered and chambered wounds, and BTES demonstrated the best healing potential. In conclusion, BTES proved to be an effective tissue-engineered construct that can promote the healing of full-thickness skin lesions. With the support of further clinical trials, this procedure could be an alternative to SSG for patients with partial- and full-thickness burns. PMID:24637651

Idrus, Ruszymah Bt Hj; Rameli, Mohd Adha bin P; Low, Kiat Cheong; Law, Jia Xian; Chua, Kien Hui; Latiff, Mazlyzam Bin Abdul; Saim, Aminuddin Bin

2014-04-01

104

In vitro susceptibilities of normal human skin fibroblasts to oncoviruses, and the decreased susceptibility to HSV of fibroblasts from untreated Hodgkin's patients.  

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Fibroblast cultures established from the skin of 56 healthy controls and 15 untreated Stages I and II Hodgkin's patients (HD) were studied in their 3rd, 4th and 5th in vitro passage with respect to transformation with Simian sarcoma virus (SSV) and SV40 and with respect to replication of herpes simplex virus (HSV) Types 1 and 2, pox virus and interferon release. Susceptibility to the 5 viruses varied independently, except for an inverse correlation between susceptibility to SSV and HSV. HD cu...

Ebbesen, P.; Vestergaard, B. F.; Ting, R.; Haahr, S.; Genner, J.; Svejgaard, A.

1981-01-01

105

Autophagic-lysosomal dysregulation downstream of cathepsin B inactivation in human skin fibroblasts exposed to UVA  

Science.gov (United States)

Recently, using 2D-DIGE proteomics we have identified cathepsin B as a novel target of UVA in human Hs27 skin fibroblasts. In response to chronic exposure to noncytotoxic doses of UVA (9.9 J/cm2, twice a week, 3 weeks), photooxidative impairment of cathepsin B enzymatic activity occurred with accumulation of autofluorescent aggregates colocalizing with lysosomes, an effect mimicked by pharmacological antagonism of cathepsin B using the selective inhibitor CA074Me. Here, we have further explored the mechanistic involvement of cathepsin B inactivation in UVA-induced autophagic-lysosomal alterations using autophagy-directed PCR expression array analysis as a discovery tool. Consistent with lysosomal expansion, UVA upregulated cellular protein levels of the lysosomal marker glycoprotein Lamp-1, and increased levels of the lipidated autophagosomal membrane constituent LC3-II were detected. UVA did not alter expression of beclin 1 (BECN1), an essential factor for initiation of autophagy, but upregulation of p62 (sequestosome 1, SQSTM1), a selective autophagy substrate, and ?-synuclein (SNCA), an autophagic protein substrate and aggresome component, was observed at the mRNA and protein level. Moreover, UVA downregulated transglutaminase-2 (TGM2), an essential enzyme involved in autophagolysosome maturation. Strikingly, UVA effects on Lamp-1, LC3-II, beclin 1, p62, ?-synuclein, and transglutaminase-2 were mimicked by CA074Me treatment. Taken together, our data suggest that UVA-induced autophagic-lysosomal alterations occur as a consequence of impaired autophagic flux downstream of cathepsin B inactivation, a novel molecular mechanism potentially involved in UVA-induced skin photodamage.

Lamore, Sarah D.; Wondrak, Georg T.

2014-01-01

106

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

Energy Technology Data Exchange (ETDEWEB)

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 T{sub 3}(0.184 x 10{sup -9} to 46 x 10{sup -9} mol/l) and were labelled with ({sup 35}S)sulphate and ({sup 3}H)glucosamine. Proteoglycans were purified by Sephadex G-50, Q-Sepharose chromatography with NaCl-gradient and Sepharose CL-6B chromatography. {sup 35}S and {sup 3}H incorporated into dermatan sulphate proteoglycan and heparan sulphate proteoglycan and {sup 3}H incorporated into hyaluronan were measured. {sup 35}S and {sup 3}H incorporation into dermatan sulphate proteoglycan was minimum at a T{sub 3} concentration of 0.184 x 10{sup -9} mol/l, and increased with increasing doses of T{sub 3} up to 46 x 10{sup -9} mol/l. {sup 35}S and {sup 3}H incorporation into heparan sulphate proteoglycan also increased with increasing-doses of T{sub 3}. {sup 3}H incorporation into hyaluranan was not influenced at all by T{sub 3}. The increased incorporation of {sup 35}S into proteoglycan in high-T{sub 3} culture reflects the increased synthesis of proteoglycan because 1. the extent of sulphation of disaccharides examined by thin-layer chromatography was not altered by T{sub 3}; 2. the specific activity of ({sup 35}S)sulphate was not influenced by T{sub 3}, and 3. T{sub 3} did not decrease the degradation rate of cell-associated proteoglycan. (author).

Shishiba, Yoshimasa; Ozawa, Yasunori; Shimizu, Taeko (Division of Endocrinology and Endocrine Research Laboratory, Toranomon Hospital (Japan)); Takeuchi, Yasuhiro; Yokoi, Noriko (Okinaka Memorial Institute for Medical Research, Akasaka, Tokyo (Japan))

1990-01-01

107

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 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)

1990-01-01

108

Autophagic-lysosomal dysregulation downstream of cathepsin B inactivation in human skin fibroblasts exposed to UVA.  

Science.gov (United States)

Recently, using 2D-DIGE proteomics we have identified cathepsin B as a novel target of UVA in human Hs27 skin fibroblasts. In response to chronic exposure to noncytotoxic doses of UVA (9.9 J cm(-2), twice a week, 3 weeks), photooxidative impairment of cathepsin B enzymatic activity occurred with accumulation of autofluorescent aggregates colocalizing with lysosomes, an effect mimicked by pharmacological antagonism of cathepsin B using the selective inhibitor CA074Me. Here, we have further explored the mechanistic involvement of cathepsin B inactivation in UVA-induced autophagic-lysosomal alterations using autophagy-directed PCR expression array analysis as a discovery tool. Consistent with lysosomal expansion, UVA upregulated cellular protein levels of the lysosomal marker glycoprotein Lamp-1, and increased levels of the lipidated autophagosomal membrane constituent LC3-II were detected. UVA did not alter expression of beclin 1 (BECN1), an essential factor for initiation of autophagy, but upregulation of p62 (sequestosome 1, SQSTM1), a selective autophagy substrate, and ?-synuclein (SNCA), an autophagic protein substrate and aggresome component, was observed at the mRNA and protein level. Moreover, UVA downregulated transglutaminase-2 (TGM2), an essential enzyme involved in autophagolysosome maturation. Strikingly, UVA effects on Lamp-1, LC3-II, beclin 1, p62, ?-synuclein, and transglutaminase-2 were mimicked by CA074Me treatment. Taken together, our data suggest that UVA-induced autophagic-lysosomal alterations occur as a consequence of impaired autophagic flux downstream of cathepsin B inactivation, a novel molecular mechanism potentially involved in UVA-induced skin photodamage. PMID:21773629

Lamore, Sarah D; Wondrak, Georg T

2012-01-01

109

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)

1991-11-05

110

Spontaneous immortalization of cultured skin fibroblasts obtained from a high-dose atomic bomb survivor  

International Nuclear Information System (INIS)

Two immortal fibroblastic cell strains (substrains) were established by culturing healthy skin cells obtained from a high-dose atomic bomb survivor (female, age 76 years, 5.14 Gy) for more than 4 years. Designated FM-U and FM-M, the two substrains share the same marker chromosome, t(5q-;6p+), but are karyotypically different, possessing hypodiploid chromosome numbers (39-43) in the former and hypertriploid (69-76) in the latter. Thus far, the two strains have passed through 117 and 156 subcultures or more than 230 and 310 cumulative population doublings, respectively, each passage requiring 4-6 days in the former and 3-4 days in the latter. In the process of immortalization, sequential rearrangement among various chromosomes presumably due to telomeric and interstitial telomeric fusions took place following the telomere shortening, particularly in the senescence and postsenescence phase cells. Of particular interest is the fact that loss of heterozygosity (LOH) of the p53 gene was demonstrated in these immortalized cell populations. In addition, the allelic patterns of the LOH of p53 differed. Further evidence indicative of infinite proliferation was demonstrated in both strains, such as the telomere elongation and the significantly low frequency of cells possessing dicentric chromosomes

1996-07-05

111

Preparation of nano/submicrometer yam and its benefits on collagen secretion from skin fibroblast cells.  

Science.gov (United States)

Nano/submicrometer-scaled yam particles have been prepared by using media-milling. The particle size of media-milled yam was confirmed by the laser light scattering method and scanning electron microscopy. Influences of media-milled yam on skin fibroblast cells (WS1) were evaluated. The size reduction did not significantly alter the proximate composition, and the presence of nanoparticles was not toxic to WS1 cells. The contents of bioactive compounds (diosgenin, stigmasterol, and ?-sitosterol) were significantly increased by media-milling, which enhanced the secretion of hTGF-? and inhibited the formation of MMP-1. Thus, the collagen secretion from WS1 was significantly increased by size reduction. Diosgenin was employed as a positive control. Nevertheless, media-milled yam exhibited greater effects on WS1 cells than diosgenin. It appeared that both diosgenin and size reduction were helpful for enhancing the secretion of collagen by WS1 cells. In addition, the irritancy of yam was eliminated by media-milling. PMID:23205552

Chiang, Lung-Hsuan; Chen, Shih-Hsin; Yeh, An-I

2012-12-19

112

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 "2"5"2Cf source. Data were fitted to a multitarget model, S/S_0 = A[1-(1-e"k"D)"N], for both X-ray and neutron dose-survival curves. A single-hit model, S/S_0 = Ae"k"D, 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)

1990-01-01

113

The effect of ursolic and oleanolic acids on human skin fibroblast cells  

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Full Text Available In this article, we look at how ursolic and oleanolic acids can be used for the purpose of quality control of natural products used in dermatocosmetology as well as of various other therapeutic preparations. Ursolic acid (UA and oleanolic acid (OA are pentacyclic triterpenes and they are constituents of many medicinal herbs. In this study, we analyzed the cytotoxic and anti-proliferative activity of OA and UA against normal human skin fibroblasts (HSF. Additionally, the scavenging activity of free radicals of both acids was analyzed. The sensitivity of cells to OA and UA activity was determined using a standard spectrophotometric (MTT assay. The free radical scavenging activity of OA and UA was measured using the DPPH• test. The F-actin cytoskeletal proteins organization was analyzed using TRITC-phalloidine fluorescent staining. The cytotoxic activity of the analyzed acids was determined using Neutral Red (NR uptake assay. Of the two isomeric compounds, UA showed a higher cytotoxic activity against HSF cells than did OA. Our investigations showed that OA, in view of its non-toxic nature, may be used as a supplementary factor for dermal preparations. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 4, pp. 664–669

Helena Donica

2012-01-01

114

Growth stimulation of human skin fibroblasts by elastin-derived peptides.  

Science.gov (United States)

Elastin-derived peptides (kappa-elastin: kappa E, mean molecular mass: 75 kDa), either coated onto plastic dishes or added to culture media (0.26 to 1.33 nM) stimulated the growth of human skin fibroblasts (HSF) strains obtained from different donors and tested at different cell passages (4 to 12). Coated 44.4 micrograms/cm2 insoluble elastin (iE) exhibited the same action; coated iE or kappa E significantly modifies the HSF morphology: after 5-6 days of culture, HSF are more elongated, and at preconfluence state, formation of HSF clusters surrounding iE were observed. Increased 3H thymidine incorporation and proliferative effect of HSF by kappa E (1.3 to 2.2 fold as compared to control cells) was observed after a lag phase period which raised with initial HSF density. Optimal proliferative effect was obtained at kappa E 8.5 10(-10) M, a value close to the dissociation constant (kD = 2.7 10(-10) M) of kappa E to HSF. Valine-glycine-valine-alanine-proline-glycine (VGVAPG), but not valine-glycine-valine (VGV) or Valine-glycine-valine-valine-glycine-alanine (VGVVGA) also significantly stimulated, optimally at 7.0 10(-10) M, HSF proliferation. It was concluded that the stimulatory influence of elastin derived peptides on HSF proliferation was mediated through a binding to plasmalemmal receptor of HSF. PMID:8821030

Kamoun, A; Landeau, J M; Godeau, G; Wallach, J; Duchesnay, A; Pellat, B; Hornebeck, W

1995-11-01

115

Androgen receptor in human skin fibroblasts. Characterization of a specific 17beta-hydroxy-5alpha-androstan-3-one-protein complex in cell sonicates and nuclei.  

Science.gov (United States)

Cultured human skin fibroblasts were shown to contain an androgen binding activity (receptor) which was heat-labile and destroyed by trypsin. Specific binding was seen after incubations of these cells with 1,2-3-H-testosterone, 1,2-3-H17beta-hydroxy-5alpha-androstan-3-one (dihydrotestosterone, DHT) and 1,2-3-H-5alpha-androstane-3alpha, 17beta-diol. This receptor had a high affinity (Kd=0,2-1.6 nM) and a high degree of specificity for DHT. It was measured as a 3-H-DHT-protein complex by gel filtration chromatography using a method which distinguishes specific from nonspecific binding. Receptor activity was distributed about equally between nuclear and extranuclear components at all times studied and was present in both compartments when cell incubations were carried out at 4 degrees and 37 degrees. Saturation analysis indicated that there were 1250-18,600 binding sites per whole cell. By sucrose gradient centrifugation the receptor had a sedimentation coefficient (S20,w) of about 4. Cells grown for 8 days without serum in the medium maintained the same levels of 3-H-DHT binding. Within 15 hours puromycin (20 mug/ml) in serum-free medium caused a 40-60 percent decrease in binding for the same cell lines. Although the highest levels of 3-H-DHT binding were observed in fibroblasts from newborn foreskin, appreciable cytosol and nuclear binding were seen in cells from forearm, neck and abdominal skin. Receptor activity was stable during prolonged culture. Fibroblasts from several skin sites from patients with the androgen insensitivity syndrome (testicular feminization) had no detectable specific DHT binding. In this study it was demonstrated that skin fibroblasts can rapidly convert testosterone to its active form, DHT, bind DHT to a specific receptor protein and transport this complex to their nuclei. Therefore this may prove to be a convenient system for studying androgen action in vitro. PMID:165598

Keenan, B S; Meyer, W J; Hadjian, A J; Migeon, C J

1975-04-01

116

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

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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 po...

Szu-Kai Chen; Chu-Hsi Hsu; Min-Lang Tsai; Rong-Huei Chen; Drummen, Gregor P. C.

2013-01-01

117

Design of vectors for efficient expression of human purine nucleoside phosphorylase in skin fibroblasts from enzyme-deficient humans  

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Purine nucleoside phosphorylase deficiency is an inherited disorder associated with a severe immune defect that is fatal. Enzyme replacement therapy is an attractive approach to treatment of this disease. To this aim the authors constructed retroviral vectors containing a human PNP cDNA and a selectable gene encoding neomycin phosphotransferase. PNP expression was controlled by either the early promoter from simian virus 40, the immediate early promoter from human cytomegalovirus, or the retroviral promoter. Cultured skin fibroblasts from two unrelated PNP-deficient patients that were infected with these vectors expressed mean PNP activities of 0.03, 0.74, and 5.9 /mu/mol/hr per mg of protein, respectively. The latter infectants had PNP activities eight times the level of 0.74 /mu/mol/hr per mg of protein observed in normal skin fibroblasts, enabling rapid metabolism of exogenous deoxyguanosine, the cytotoxic metabolite that accumulates in the plasma of PNP-deficient patients. These experiments indicate that viral long terminal repeat was the strongest promoter for expression of PNP and suggest the potential of human skin fibroblasts as vehicles for therapeutic gene expression.

Osborne, W.R.A.; Miller, A.D.

1988-09-01

118

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

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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

119

Fibroblast cell line establishment, cryopreservation and interspecies embryos reconstruction in red panda ( Ailurus fulgens).  

Science.gov (United States)

In evolution, the red panda (Ailurus fulgens) plays a pivotal role in the higher level phylogeny of arctoides carnivore mammals. The red panda inhabits certain Asian countries only and its numbers are decreasing. Therefore, the development of feasible ways to preserve this species is necessary. Genetic resource cryopreservation and somatic cell nuclear transfer (SCNT) have been used extensively to rescue this endangered species. The present study describes the establishment, for the first time, of a red panda ear fibroblast cell line, which was then cryopreserved, thawed and cultured. Through micromanipulation, interspecies embryos were reconstructed using the cryopreserved-thawed fibroblasts of the red panda as the donor and rabbit oocytes as recipients. A total of 194 enucleated rabbit oocytes were reconstructed with red panda ear fibroblasts; enucleated oocytes were activated without fusion as the control. The results show that the fibroblast cell line was established successfully by tissue culture and then cryopreserved in liquid nitrogen. Supplementation with 20% fetal bovine serum and 8% dimethyl sulphoxide in basic medium facilitated the cryopreservation. The interspecies embryos were successfully reconstructed. The cleavage, morulae and blastocyst rates after in vitro culture were 71, 47 and 23% (31/194), respectively. This study indicated that a somatic cell line could be established and cryopreserved from red panda and that rabbit cytoplast supports mitotic cleavage of the red panda karyoplasts and is capable of reprogramming the nucleus to achieve blastocysts. PMID:19232145

Tao, Yong; Liu, Jianming; Zhang, Yunhai; Zhang, Meiling; Fang, Junshun; Han, Wei; Zhang, Zhizhong; Liu, Ya; Ding, Jianping; Zhang, Xiaorong

2009-05-01

120

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

Science.gov (United States)

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” silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-?, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-?, IL-1? and nitric oxide) production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair.

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

2013-01-01

 
 
 
 
121

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 tropoelastin accumulation, and altered proteoglycans. (author)

2002-02-01

122

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)

1986-01-01

123

Glucocorticoid coordinate regulation of type I procollagen gene expression and procollagen DNA-binding proteins in chick skin fibroblasts  

International Nuclear Information System (INIS)

Nuclei were isolated from control and dexamethasone-treated (2 h) embryonic chick skin fibroblasts and transcribed in vitro. Nuclei isolated from dexamethasone-treated fibroblasts transcribed less pro?1(I) and pro?2(I) mRNAs but not ?-actin mRNA. Fibroblasts receiving dexamethasone and [5,6-3H]uridine also demonstrated decreased synthesis of nuclear type I procollagen mRNAs but not ?-actin mRNA. In fibroblasts treated with cycloheximide the newly synthesized nuclear type I procollagen mRNA species were markedly decreased. An enhanced inhibitory effect was observed when fibroblasts were treated with cycloheximide plus dexamethasone. Since the studies above demonstrate that active protein synthesis is required to maintain the constitutive expression of the type I procollagen genes, the authors determined if glucocorticoids regulate DNA-binding proteins with sequence specificity for the ?2(I) procollagen gene. Nuclear protein blots were probed with the 32P-end-labeled pBR322 vector DNA and 32P-end-labeled ?2(I) procollagen promoter containing DNA. Nonhistone proteins remained bound to labeled DNA at stringency washes of 0.05 and 0.1 M NaCl. As the ionic strength was increased to 0.2 and 0.3 M NaCl, the nonhistone-protein DNA binding was preferentially lost. Only the low molecular weight proteins remained bound to labeled DNA at the highest ionic strength, indicating nonspecific binding of these nuclear proteins. Dexamethasone treatment resulted in an increase of binding of nonhistone proteins to vector- and promoter-labeled DNAs over that observed in control fibroblasts at stringency washes of 0.05 and 0.1 M NaCl and to a lesser extent at 0.2 M NaCl. The binding specificities of nonhistone proteins for the ?2(I) procollagen promoter containing DNA were calculated

1988-04-19

124

Methylmalonic and propionic acidemias: lipid profiles of normal and affected human skin fibroblasts incubated with [1-14C]propionate  

International Nuclear Information System (INIS)

Normal human skin fibroblasts and those from methylmalonic acidemia and propionic acidemia patients were grown in culture. Following incubation with [1-14C]propionate, the major lipid classes in the cells were separated by thin layer chromatography and isolated fractions analyzed by radio gas chromatography for the presence of odd-numbered long-chain fatty acids; the pattern of even-numbered long-chain fatty acids was obtained also. Normal fibroblasts incorporated a small percentage of propionate into odd-numbered fatty acids which were present in all lipids studied. The abnormal cells incorporated a larger amount while maintaining the characteristic ratios of odd-numbered fatty acids found in the normal line. Most of the radioactivity was associated with phospholipids which are the predominant constituents of cell membranes. A characteristic C15/C17 ratio was found for different phospholipids and the triglyceride fraction; pentadecanoic acid was the principal odd-numbered fatty acid utilized in the assembly of complex lipids. Compared to even-numbered long-chain fatty acids the absolute amount of odd-numbered fatty acids was low (1-2%), even in affected cells. An unusual polar lipid fraction was isolated in the course of the study. In the normal cell it contained several unlabeled eicosanoids which were missing from the same fraction of both affected cell lines

1986-01-01

125

Impaired colony-forming ability following ? irradiation of skin fibroblasts from tuberous scierosis patients  

International Nuclear Information System (INIS)

The radiosensitivity of cultured dermal fibroblasts from human subjects afflicted with tuberous sclerosis (TS), a hereditary neurocutaneous syndrome, was assessed by assaying loss of colony-forming ability in response to acute ?-ray exposure. Related to control strains from clinically normal donors, three cell lines (GM1635, GM1643, GM2333) from two affected patients displayed enhanced sensitivity to inactivation by 60Co ?-ray treatment, whether administered oxically (air-saturated) or hypoxically (N2-gassed); a fourth strain (GM1644) from a third patient exhibited normal radiosensitivity under both treatment conditions. The post-?-irradiaton colony-forming ability of the three hypersensitive TS strains was intermediate between that of normal controls and that of strains from patients inheriting the radiotherapy-sensitive neurovascular disorder ataxia telangiectasia. The variability in the radioresponse of the TS stains (three sensitive and one normal) is not surprising, considering the widely recognized clinical heterogeneity in the disease. Our findings, aside from providing a laboratory marker for early (possible presymptomatic) detection of persons at high risk for TS, may lead to a better understanding of the origin and progressive development of this multifaceted syndrome

1982-01-01

126

Relationship between in vitro radiosensitivity of normal human skin fibroblasts and the occurrence of late normal tissue reactions after radiotherapy  

International Nuclear Information System (INIS)

Late complications in normal tissues are limiting for the doses that can be administered during clinical radiotherapy. Awareness of these complications, and comprehension of the underlying biological mechanisms, is extremely important to improve cancer treatment. Fibrosis is one of the most critical injuries to radiotherapy. It varies significantly among patients despite of identical treatments. The large patient-to-patient variability of normal tissue sections to clinical radiation can possibly be accounted for by the considerable individual variation in cellular radiosensitivity of normal human fibroblasts, as shown in vitro. The purpose of the present investigation has been to analyze individual cellular radiosensitivity of normal human skin fibroblasts, as measured in a colony-forming assay, and the relationship to the occurrence of subcutaneous fibrosis after radiotherapy for breast cancer. (au) 97 refs

1995-01-01

127

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

Science.gov (United States)

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. PMID:21205999

Chandrasekaran, Arun Richard; Venugopal, J; Sundarrajan, S; Ramakrishna, S

2011-02-01

128

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.

2011-02-01

129

Synthesis of glycosaminoglycans by human skin fibroblasts cultured on collagen gels.  

Science.gov (United States)

A comparison has been made of the synthesis of glycosaminoglycans by human skin fibroblasts cultured on plastic or collagen gel substrata. Confluent cultures were incubated with [3H]glucosamine and Na235SO4 for 48h. Radiolabelled glycosaminoglycans were then analysed in the spent media and trypsin extracts from cells on plastic and in the medium, trypsin and collagenase extracts from cells on collagen gels. All enzyme extracts and spent media contained hyaluronic acid, heparan sulphate and dermatan sulphate. Hyaluronic acid was the main 3H-labelled component in media and enzyme extracts from cells on both substrata, although it was distributed mainly to the media fractions. Heparan sulphate was the major [35S]sulphated glycosaminoglycan in trypsin extracts of cells on plastic, and dermatan sulphate was the minor component. In contrast, dermatan sulphate was the principal [35S]sulphated glycosaminoglycan in trypsin and collagenase extracts of cells on collagen gels. The culture substratum also influenced the amounts of [35S]sulphated glycosaminoglycans in media and enzyme extracts. With cells on plastic, the medium contained most of the heparan sulphate (75%) and dermatan sulphate (> 90%), whereas the collagenase extract was the main source of heparan sulphate (60%) and dermatan sulphate (80%) from cells on collagen gels; when cells were grown on collagen, the medium contained only 5-20% of the total [35S]sulphated glycosaminoglycans. Depletion of the medium pool was probably caused by binding of [35S]sulphated glycosaminoglycans to the network of native collagen fibres that formed the insoluble fraction of the collagen gel. Furthermore, cells on collagen showed a 3-fold increase in dermatan sulphate synthesis, which could be due to a positive-feedback mechanism activated by the accumulation of dermatan sulphate in the microenvironment of the cultured cells. For comparative structural analyses of glycosaminoglycans synthesized on different substrata labelling experiments were carried out by incubating cells on plastic with [3H]glucosamine, and cells on collagen gels with [14C]glucosamine. Co-chromatography on DEAE-cellulose of mixed media and enzyme extracts showed that heparan sulphate from cells on collagen gels eluted at a lower salt concentration than did heparan sulphate from cells on plastic, whereas with dermatan sulphate the opposite result was obtained, with dermatan sulphate from cells on collagen eluting at a higher salt concentration than dermatan sulphate from cells on plastic. These differences did not correspond to changes in the molecular size of the glycosaminoglycan chains, but they may be caused by alterations in polymer sulphation. PMID:7470051

Gallagher, J T; Gasiunas, N; Schor, S L

1980-08-15

130

Dexamethasone regulation of glycosaminoglycan synthesis in cultured human skin fibroblasts. Similar effects of glucocorticoid and thyroid hormones.  

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The effects of dexamethasone on glycosaminoglycan accumulation were examined in confluent human skin fibroblasts in vitro. The glucocorticoid consistently inhibited the incorporation of either [3H]acetate or [3H]glucosamine into hyaluronate when added to culture medium 72 h before harvest. This effect was half-maximal at approximately 1 nM and maximal at 5-10 nM. Inhibition occurred within 5 h of hormone addition and was near maximal by 25 h. 11 alpha-hydrocortisone (10 nM), deoxycorticostero...

Smith, T. J.

1984-01-01

131

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

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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

132

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-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

1991-02-15

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

1988-01-01

134

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

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[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

135

Extracellular Matrix Modulates Morphology, Growth, Oxidative Stress Response and Functionality of Human Skin Fibroblasts during Aging In Vitro  

DEFF Research Database (Denmark)

The Hayflick system of cellular aging and replicative senescence in vitro has been used widely in both basic and applied research in biogerontology. The state of replicative senescence is generally considered to be irreversible, but is modifiable by genetic and environmental manipulations. Some recent observations indicate that replicative lifespan, senescence and functionality of cells in vitro can be significantly affected by the quality of the extra cellular matrix (ECM). Following up on those reports, here we show that using the ECM prepared from early passage young cells, partial rejuvenation of serially passaged human facial skin fibroblasts was possible in pre-senescent middle-aged cells, but not in fully senescent late passage cells. ECM from young cells improved the appearance, viability, stress tolerance and wound healing ability of skin fibroblasts. Furthermore, young ECM modulated the oxidative stress response transcription factor Nrf-2 and its downstream effector haem-oxygenase (HO-1), possibly through the amelioration of the environmental stress induced by the plastic surface of the culturing flasks. Therefore, it is important to consider the role of ECM in modulating the response of cells both for mechanistic understanding of cellular senescence and while testing for potential aging interventions.

Rattan, Suresh; Jørgensen, Peter

2014-01-01

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

Abnormal cytoskeletal protein expression in cultured skin fibroblasts from type 1 diabetes mellitus patients with nephropathy: A proteomic approach.  

Science.gov (United States)

Diabetic nephropathy (DN) develops in about 40% of insulin-dependent type 1 diabetes mellitus (T1DM) patients, and is associated not only with diabetes duration and metabolic control, but also with a genetic predisposition. Constitutive alterations of cytoskeletal proteins may play a role in the development of DN. We investigated the expression of these proteins in cultured skin fibroblasts, obtained from long-term T1DM patients with and without DN but comparable metabolic control, and from matched healthy subjects, by means of 2-DE electrophoresis and MS-MALDI analyses. In T1DM with DN, compared to the other two groups, quantitative analyses revealed an altered expression of 17 spots (p<0.05-p<0.01), corresponding to 12 unique proteins. In T1DM with DN, beta-actin and three isoforms of tubulin beta-2 chain, tropomodulin-3, and LASP-1 were decreased, whereas two tubulin beta-4 chain isoforms, one alpha actinin-4 isoform, membrane-organizing extension spike protein (MOESIN), FLJ00279 (corresponding to a fragment of myosin heavy chain, non-muscle type A), vinculin, a tropomyosin isoform, and the macrophage capping protein were increased. A shift in caldesmon isoforms was also detected. These results demonstrate an association between DN and the constitutive expression of cytoskeleton proteins in cultured skin fibroblasts from T1DM with DN, which may retain pathophysiologycal implications. PMID:21136853

Millioni, Renato; Iori, Elisabetta; Puricelli, Lucia; Arrigoni, Giorgio; Vedovato, Monica; Trevisan, Roberto; James, Peter; Tiengo, Antonio; Tessari, Paolo

2008-04-01

138

[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

139

Supplementation with a complex of active nutrients improved dermal and epidermal characteristics in skin equivalents generated from fibroblasts from young or aged donors.  

Science.gov (United States)

Cultured skin equivalent (SE, Mimeskin) was generated by co-culturing skin fibroblasts and keratinocytes on a collagen-glycosaminoglycan-chitosan dermal substrate. In order to examine donor age effect, fibroblasts from 19- (young) or 49- (aged) year-old females were used. Culture medium was supplemented with nutrients complex containing soy extract, tomato extract, grape seed extract, white tea extract, sodium ascorbate, tocopherol acetate, zinc gluconate and BioMarine complex. Epidermal and dermal structure and composition were examined after 42 and 60 days of culture. In untreated samples, SE generated from young fibroblasts was superior to SE from aged fibroblasts in all characteristics. Those include number and regularity of keratinocyte layers, number of keratinocytes expressing proliferation marker Ki67, content of collagen type I, fibrillin-1, elastin, and SE lifespan. Effects of nutritional supplementation were observed in SE from both young and aged fibroblasts, however, those effects were more pronounced in SE from aged fibroblasts. In epidermis, the treatment increased number of keratinocyte layers and delayed epidermal senescence. The number of cells expressing Ki67 was nine folds higher than those of controls, and was similar to that of young cell SE. In dermis, the treatment increased mRNA synthesis of collagen I, fibrillin-1 and elastin. In conclusion, skin cell donor age had major important effect on formation of reconstructed SE. Imperfections in epidermal and dermal structure and composition as well as life span in SE from aged cells can be improved by supplementation with active nutrients. PMID:17028931

Lacroix, Sophie; Bouez, Charbel; Vidal, Sandrine; Cenizo, Valérie; Reymermier, Corinne; Justin, Virginie; Vicanová, Jana; Damour, Odile

2007-04-01

140

Novel collagen/gelatin scaffold with sustained release of basic fibroblast growth factor: clinical trial for chronic skin ulcers.  

Science.gov (United States)

Chronic skin ulcers such as diabetic ulcers and venous leg ulcers are increasing and are a costly problem in healthcare. We have developed a novel artificial dermis, collagen/gelatin sponge (CGS), which is capable of sustained release of basic fibroblast growth factor (bFGF) for more than 10 days. The objective of this study was to investigate the safety and efficacy of CGS impregnated with bFGF in the treatment of chronic skin ulcers. Patients with chronic skin ulcers that had not healed in at least 4 weeks were treated with CGS impregnated with bFGF at 7 or 14 ?g/cm(2) after debridement, and the wound bed improvement was assessed 14 days after application. Wound bed improvement was defined as a granulated and epithelialized area on day 14 with a proportion to the baseline wound area after debridement of 50% or higher. The wound area, the wound area on day 14, and the granulation area on day 14 were independently measured by blinded reviewers in a central review using digital images of wounds taken with a calibrator. Patients were followed up until 28 days after application to observe the adverse reactions related to the application of CGS. From May 2010 to June 2011, 17 patients were enrolled and, in 16 patients, the wound bed improved. Among the randomized patients in step 2, no significant difference was seen between the low-dose group and the high-dose group. No serious adverse reactions were observed. Adverse reactions with a clear causal relationship to the study treatment were mild and patients quickly recovered from them. This study is the first-in-man clinical trial of CGS and showed the safety and efficacy of CGS impregnated with bFGF in the treatment of chronic skin ulcers. This combination therapy could be a promising therapy for chronic skin ulcers. PMID:23541061

Morimoto, Naoki; Yoshimura, Kenichi; Niimi, Miyuki; Ito, Tatsuya; Aya, Rino; Fujitaka, Junpei; Tada, Harue; Teramukai, Satoshi; Murayama, Toshinori; Toyooka, Chikako; Miura, Kazumi; Takemoto, Satoru; Kanda, Norikazu; Kawai, Katsuya; Yokode, Masayuki; Shimizu, Akira; Suzuki, Shigehiko

2013-09-01

 
 
 
 
141

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

1984-01-01

142

Efficient retrovirus-mediated transfer and expression of a human adenosine deaminase gene in diploid skin fibroblasts from an adenosine deaminase-deficient human  

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Skin fibroblasts might be considered suitable recipients for therapeutic genes to cure several human genetic diseases; however, these cells are resistant to gene transfer by most methods. The authors studied the ability of retroviral vectors to transfer genes into normal human diploid skin fibroblasts. Retroviruses carrying genes for neomycin or hygromycin B resistance conferred drug resistance to greater than 50% of the human fibroblasts after a single exposure to virus-containing medium. This represents at least a 500-fold increase in efficiency over other methods. Transfer was achieved in the absence of helper virus by using amphotropic retrovirus-packaging cells. A retrovirus vector containing a human adenosine deaminase (ADA) cDNA was constructed and used to infect ADA/sup -/ fibroblasts from a patient with ADA deficiency. The infected cells produced 12-fold more ADA enzyme than fibroblasts from normal individuals and were able to rapidly metabolize exogenous deoxyadenosine and adenosine, metabolites that accumulate in plasma in ADA-deficient patients and are responsible for the severe combined immunodeficiency in these patients. These experiments indicate the potential of retrovirus-mediated gene transfer into human fibroblasts for gene therapy.

Palmer, T.D.; Hock, R.A.; Osborne, W.R.A.; Miller, A.D.

1987-02-01

143

Pseudohypophosphatasia: aberrant localization and substrate specificity of alkaline phosphatase in cultured skin fibroblasts.  

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We explored the biochemical basis for the disorder pseudohypophosphatasia (PsHYPT) in one patient by examining the substrate specificity and localization of alkaline phosphatase (ALP) in cultured dermal fibroblasts. Despite substantial ALP activity, in cell homogenates, toward the artificial substrate 4-methyl-umbelliferyl phosphate (4-MUP), there was a marked deficiency in ALP activity toward the natural substrates pyridoxal 5'-phosphate (PLP) and phosphoethanolamine (PEA), indicating altere...

Fedde, K. N.; Cole, D. E.; Whyte, M. P.

1990-01-01

144

Basic Fibroblast Growth Factor and Ultraviolet B Transform Melanocytes in Human Skin  

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Ultraviolet (UV) light is an epidemiological risk factor for melanoma, but its specific contribution to melanoma induction is not known. The first critical step of melanoma development, ie, the uncontrolled proliferation of melanocytes, may be induced by a combination of UV damage and an imbalance of growth factor production by cells in the immediate area of the melanocyte. Among several candidates, basic fibroblast growth factor (bFGF) is the major autocrine growth factor in melanoma and ass...

Berking, Carola; Takemoto, Richelle; Satyamoorthy, Kapaettu; Elenitsas, Rosalie; Herlyn, Meenhard

2001-01-01

145

Molecular species of phospholipids with very long chain fatty acids in skin fibroblasts of Zellweger syndrome.  

Science.gov (United States)

The ratio of C 26:0/C 22:0 fatty acids in patient lipids is widely accepted as a critical clinical criterion of peroxisomal diseases, such as Zellweger syndrome and X-linked adrenoleukodystrophy (X-ALD). However, phospholipid molecular species with very long chain fatty acids (VLCFA) have not been precisely characterized. In the present study, the structures of such molecules in fibroblasts of Zellweger syndrome and X-ALD were examined using LC-ESI-MS/MS analysis. In fibroblasts from Zellweger patients, a large number of VLCFA-containing molecular species were detected in several phospholipid classes as well as neutral lipids, including triacylglycerol and cholesteryl esters. Among these lipids, phosphatidylcholine showed the most diversity in the structures of VLCFA-containing molecular species. Some VLCFA possessed longer carbon chains and/or larger number of double bonds than C 26:0-fatty acid (FA). Similar VLCFA were also found in other phospholipid classes, such as phosphatidylethanolamine and phosphatidylserine. In addition, VLCFA-containing phospholipid species showed some differences among fibroblasts from Zellweger patients. It appears that phospholipids with VLCFA, with or without double bonds, as well as C 26:0-FA might affect cellular functions, thus leading to the pathogenesis of peroxisomal diseases, such as Zellweger syndrome and X-ALD. PMID:24122089

Hama, Kotaro; Nagai, Toru; Nishizawa, Chiho; Ikeda, Kazutaka; Morita, Masashi; Satoh, Noriko; Nakanishi, Hiroki; Imanaka, Tsuneo; Shimozawa, Nobuyuki; Taguchi, Ryo; Inoue, Keizo; Yokoyama, Kazuaki

2013-12-01

146

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

147

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.

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

2013-01-01

148

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

149

The Impressive Healing Power of Autologous Fibroblasts Isolated from Early Cultures of Skin Biopsies for the Treatment of Diabetic Foot Ulcers: Preliminary Results Regarding 2 Cases  

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Aims: The purpose of this study was to investigate whether we can quickly, effectively, and with relatively low cost, heal long-standing (>8 months) diabetic foot ulcers using autologous skin fibroblasts. Place and Duration of Study: Immunology & National Histocompatibility Department and 2nd Department of Surgery, ‘G. Gennimatas’ General Hospital, ‘Demetrios Voyatzoglou’ Diabetic Foot Clinic, ‘A. Fleming’ General Hospital, Department of Biology, ...

2013-01-01

150

Abnormal sensitivity of skin fibroblasts from familial polyposis patients to DNA alkylating agents  

International Nuclear Information System (INIS)

Fibroblast cell strains derived from different patients all afflicted with genetic predisposing to the development of intestinal polyposis and cancer were tested for their sensitivity to the lethal effects of the DNA alkylating agents methylmethanesulfonate (MMS), ethyl methanesulfonate, N-methyl-N'-nitro-N-nitrosoguanidine, and 4-nitroquinoline 1-oxide. The genetic syndromes studied were: (a) adenomatosis of the colon and rectum only, an autosomal dominant trait; (b) Turcot's syndrome, a rare autosomal recessive polyposis syndrome also characterized by central nervous system tumors; and (c) Gardner's syndrome, an autosomal dominant syndrome which, in addition to intestinal polyposis, is also clinically characterized by osteomas and soft tissue tumors. Fibroblasts from a patient with Turcot's syndrome were hypersensitive to MMS, having a D0 value of 0.24 mM (p less than 0.01) versus the normal average D0 of 0.36 mM and a D10 value of 0.95 mM (p less than 0.01) compared with the normal average value of 1.3 mM. Fibroblasts from the Gardner's syndrome proband were moderately sensitive to MMS, ethyl methanesulfonate, and N-methyl-N'-nitro-N-nitrosoguanidine due to significant differences of D10 values of 0.60 mM (p less than 0.01), 15 mM (p less than 0.01), and 4.8 microM (p less than 0.025), respectively, versus the normal average values of 1.3 mM, 28 mM, and 9.4 microM. Fibroblasts from the clinically affected Gardner's syndrome daughter of the proband were significantly more sensitive to MMS treatment, D0 of 0.22 mM (p less than 0.01) versus the normal average D0 of 0.36 mM and a D10 of 0.97 mM (p less than 0.01) versus the normal average. This differential sensitivity to the several DNA alkylating agents suggests that different mechanisms of hypersensitivity to these chemicals may be associated with fibroblasts from the various forms of familial polyposis

1982-01-01

151

Abnormal sensitivity of skin fibroblasts from familial polyposis patients to DNA alkylating agents  

Energy Technology Data Exchange (ETDEWEB)

Fibroblast cell strains derived from different patients all afflicted with genetic predisposing to the development of intestinal polyposis and cancer were tested for their sensitivity to the lethal effects of the DNA alkylating agents methylmethanesulfonate (MMS), ethyl methanesulfonate, N-methyl-N'-nitro-N-nitrosoguanidine, and 4-nitroquinoline 1-oxide. The genetic syndromes studied were: (a) adenomatosis of the colon and rectum only, an autosomal dominant trait; (b) Turcot's syndrome, a rare autosomal recessive polyposis syndrome also characterized by central nervous system tumors; and (c) Gardner's syndrome, an autosomal dominant syndrome which, in addition to intestinal polyposis, is also clinically characterized by osteomas and soft tissue tumors. Fibroblasts from a patient with Turcot's syndrome were hypersensitive to MMS, having a D0 value of 0.24 mM (p less than 0.01) versus the normal average D0 of 0.36 mM and a D10 value of 0.95 mM (p less than 0.01) compared with the normal average value of 1.3 mM. Fibroblasts from the Gardner's syndrome proband were moderately sensitive to MMS, ethyl methanesulfonate, and N-methyl-N'-nitro-N-nitrosoguanidine due to significant differences of D10 values of 0.60 mM (p less than 0.01), 15 mM (p less than 0.01), and 4.8 microM (p less than 0.025), respectively, versus the normal average values of 1.3 mM, 28 mM, and 9.4 microM. Fibroblasts from the clinically affected Gardner's syndrome daughter of the proband were significantly more sensitive to MMS treatment, D0 of 0.22 mM (p less than 0.01) versus the normal average D0 of 0.36 mM and a D10 of 0.97 mM (p less than 0.01) versus the normal average. This differential sensitivity to the several DNA alkylating agents suggests that different mechanisms of hypersensitivity to these chemicals may be associated with fibroblasts from the various forms of familial polyposis.

Barfknecht, T.R.; Little, J.B.

1982-04-01

152

Shell Extracts from the Marine Bivalve Pecten maximus Regulate the Synthesis of Extracellular Matrix in Primary Cultured Human Skin Fibroblasts  

Science.gov (United States)

Mollusc shells are composed of more than 95% calcium carbonate and less than 5% of an organic matrix consisting mostly of proteins, glycoproteins and polysaccharides. Previous studies have elucidated the biological activities of the shell matrices from bivalve molluscs on skin, especially on the expression of the extracellular matrix components of fibroblasts. In this work, we have investigated the potential biological activities of shell matrix components extracted from the shell of the scallop Pecten maximus on human fibroblasts in primary culture. Firstly, we demonstrated that shell matrix components had different effects on general cellular activities. Secondly, we have shown that the shell matrix components stimulate the synthesis of type I and III collagens, as well as that of sulphated GAGs. The increased expression of type I collagen is likely mediated by the recruitment of transactivating factors (Sp1, Sp3 and human c-Krox) in the ?112/?61 bp COL1A1 promoter region. Finally, contrarily to what was obtained in previous works, we demonstrated that the scallop shell extracts have only a small effect on cell migration during in vitro wound tests and have no effect on cell proliferation. Thus, our research emphasizes the potential use of shell matrix of Pecten maximus for dermo-cosmetic applications.

Latire, Thomas; Legendre, Florence; Bigot, Nicolas; Carduner, Ludovic; Kellouche, Sabrina; Bouyoucef, Mouloud; Carreiras, Franck; Marin, Frederic; Lebel, Jean-Marc; Galera, Philippe; Serpentini, Antoine

2014-01-01

153

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

1988-01-01

154

DNA double strand breaks in fibroblast cell lines from non-Hodgkin's lymphoma patients showing increased sensitivity to chronic gamma irradiation  

International Nuclear Information System (INIS)

Cultured skin fibroblast cell lines from two non-Hodgkin's lymphoma patients (NHL) and a normal subject were studied for cell killing, chromosomal aberrations (breaks, translocations, dicentrics and rings) and DNA double strand breaks (dsbs) following chronic gamma irradiation. Compared to the cell line from the normal donor, the NHL patients' fibroblasts showed enhanced radiosensitivity for both cell survival and chromosomal aberrations. While spontaneous breaks were observed in both normal and patients' cells, spontaneous translocations and radiation-induced dicentrics and rings were found only in the latter. Radiation-induced DNA double-strand breaks (dsb) were determined by CHEF electrophoresis. After chronic irradiation with gamma rays the fraction of residual dsb was significantly increased from 1.4% in controls to 1.9% in the NHL cell lines. These data, thus suggest that the cellular and chromosomal sensitivity to chronic irradiation observed in NHL patients may be due to a deficiency in the repair of a small fraction of DNA double strand breaks. (author)

1991-05-01

155

The role of fibroblast growth factor receptor 2b in skin homeostasis and cancer development  

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The epithelial isoform of fibroblast growth factor receptor 2 (Fgfr2b) is essential for embryogenesis, and Fgfr2b-null mice die at birth. Using Cre-Lox transgenics to delete Fgfr2b in cells expressing keratin 5, we show that mice lacking epidermal Fgfr2b survive into adulthood but display striking abnormalities in hair and sebaceous gland development. Epidermal hyperthickening develops with age, and 10% of mutant mice develop spontaneous papillomas, demonstrating the role of Fgfr2b in post-na...

Grose, Richard; Fantl, Vera; Werner, Sabine; Chioni, Athina-myrto; Jarosz, Monika; Rudling, Robert; Cross, Barbara; Hart, Ian R.; Dickson, Clive

2007-01-01

156

Specific toxicity of aphidicolin to ultraviolet-irradiated excision proficient human skin fibroblasts  

International Nuclear Information System (INIS)

Aphidicolin, a specific inhibitor of the eucaryotic ? polymerase, has been employed to study the role of this enzyme in repair of potentially lethal damage (PLD) induced by far u.v. (254 nm) radiation in normal and repair defective primary human fibroblasts. There is a strong concentration dependent specific toxicity to cells treated with a fluence of 6 Jm-2 of far-u.v. radiation and incubated with aphidicolin for 2 days over the concentration range 0.0025-2.5 ?g/ml. A similar effect is seen with a xeroderma pigmentosum (XP) variant (excision proficient) strain but there is no specific toxicity to u.v irradiated excision deficient XP cells of complementation group A. Inactivation of irradiated excision proficient fibroblasts is rapid over the first 6 h of aphidicolin (1 ?g/ml) treatment but the reaction takes 2 days or longer to complete depending on the u.v. dose. These results demonstrate that the apparent uncoupling of excision repair seen previously by other investigators prevents repair of PLD and is lethal to the cells. (author)

1983-01-01

157

Spontaneous mutation rates of tumorigenic and nontumorigenic Chinese hamster embryo fibroblast cell lines.  

Science.gov (United States)

The genomic stability of a series of nontumorigenic, tumorigenic, and tumor-derived Chinese hamster embryo fibroblastic (CHEF) cell lines was compared by examining their rates of spontaneous mutation at the hypoxanthine-guanine phosphoribosyl transferase (hprt) locus, using thioguanine resistance for selection of mutants. The spontaneous mutation rates were 1.1 x 10(-6) mutations/cell/generation in the non-tumor-forming CHEF/18 cell line and 4.9 x 10(-6) in the tumorigenic CHEF/16 cells. Three tumorigenic and tumor-derived CHEF cell lines derived from CHEF/18 (J132 3-2 T3L, focus 2, focus 3) and two lines (16-2 Tuk 4 and 204 Bu50 Tuk 2) derived from CHEF/16 were chosen on the basis of their karyotypes, which demonstrated a considerable level of chromosomal rearrangement. Mutation rates of four of these five lines ranged from 1.2 x 10(-6) to 8.9 x 10(-6) mutations per cell per generation. Only the fifth line, 16-2 Tuk 4, showed a significantly elevated rate of mutation as compared with the nontumorigenic CHEF/18 cell line. Thus, we have found no simple correlation between spontaneous mutation rate and the malignant phenotype, and we conclude that mutation rate per se is not a sensitive index of malignancy. In addition, we have compared three methods of calculating mutation rate and find that they rank the cell lines in the same order, but each stresses a different aspect of the distribution and therefore produces different estimates of the mutation rate. PMID:2720691

Kaden, D; Gadi, I K; Bardwell, L; Gelman, R; Sager, R

1989-06-15

158

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 mustard treated virus. (Auth.)

1978-06-01

159

Genetic heterogeneity of inflammatory response and skin tumorigenesis in phenotypically selected mouse lines.  

Science.gov (United States)

Non-inbred AIR (AIRmax, AIRmin) and Car (Car-S, Car-R) mouse lines were generated from the same eight inbred mice through bidirectional selective breeding for acute inflammatory response and for susceptibility to two-stage skin tumorigenesis, respectively. Because AIR lines also showed a differential predisposition to skin tumorigenesis and Car lines differed in the extent of inflammatory response, we carried out genome-wide association studies using SNP arrays to identify the genetic elements affecting skin tumor susceptibility and inflammatory response in AIR and Car lines. We found that the phenotypic outcome reflects a specific genetic profile in each mouse line, suggesting that distinct genetic elements, selected by differential genetic drifts, and exerting pleiotropic effects in each mouse population, control the skin tumor susceptibility and inflammatory response phenotypes. These findings point to the complex link between skin tumor susceptibility and inflammatory response in mice. PMID:20307927

Galvan, Antonella; Vorraro, Francisca; Cabrera, Wafa Hanna Koury; Ribeiro, Orlando Garcia; Pazzaglia, Simonetta; Mancuso, Mariateresa; Zolin, Anna; Milani, Silvano; Saran, Anna; Ibañez, Olga M; Dragani, Tommaso A

2010-09-01

160

Replacement of primary chicken embryonic fibroblasts (CEF) by the DF-1 cell line for detection of avian leucosis viruses  

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International regulations prescribe that the absence of avian leucosis viruses (ALV) in avian live virus vaccines has to be demonstrated. Primary chicken embryo fibroblasts (CEF) from special SPF chicken lines are normally used for detection of ALV. The suitability of the DF-1 cell line for ALV-detection, as alternative for primary CEF, was studied in three types of experiments: (1) in titration experiments without cell passage, (2) in experiments with passages in cell cultures according to E...

Maas, R.; Zoelen-bos, D. J.; Oei, H. L.; Claassen, I. J. T. M.

2006-01-01

 
 
 
 
161

Photodynamic effects of zinc oxide nanowires in skin cancer and fibroblast.  

Science.gov (United States)

Cytotoxic effects of zinc oxide (ZnO) nanomaterials, individual and conjugated with a photosensitizer (protoporphyrin IX), were studied in the presence and absence of ultraviolet light exposure (240 nm of light wavelength for a very short time exposure) in cell cultures of human normal and cancerous skin models. Zinc Oxide nanowires (ZnO NWs) were grown on the capillary tip and conjugated with protoporphyrin IX (PpIX). This coated tip was used as tool/pointer for intracellular drug delivery protocol in suggested normal as well as carcinogenic cellular models. After true delivery of optimal drug, the labelled biological model was irradiated with UV-A, which led to a loss of mitochondrial membrane potential, as tested by neutral red assay (NRA). PMID:24338134

Fakhar-e-Alam, Muhammad; Kishwar, S; Willander, M

2014-05-01

162

Collagen synthesis in human skin fibroblasts is stimulated by a stable form of ascorbate, 2-O-alpha-D-glucopyranosyl-L-ascorbic acid.  

Science.gov (United States)

We evaluated the effect of 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G) on collagen synthesis in cultured human skin fibroblasts and on proliferation of fibroblasts. At concentrations of 0.1-0.5 mmol/L, AA-2G effectively stimulated collagen synthesis with an effectiveness comparable to that of L-ascorbic acid. On the other hand, 6-O-alpha-D-glucopyranosyl-L-ascorbic acid showed a weak effect. The stimulation of collagen synthesis by AA-2G was attenuated by the addition of a collagen synthesis inhibitor, L-azetidine 2-carboxylic acid, in a dose-dependent manner. In addition, AA-2G-induced stimulation of collagen synthesis could be completely inhibited by the addition of castanospermine, an inhibitor of neutral alpha-glucosidase. Relatively high alpha-glucosidase activity, which would contribute to release of ascorbic acid from AA-2G, could be detected in the lysate of cultured fibroblasts. The stimulatory activity of AA-2G on collagen synthesis was observed after 5 d in culture, whereas L-ascorbic acid tended to lose its stimulatory activity. Continuous supplementation of AA-2G (0.25 mmol/L) to culture medium for 24 d enhanced the cell growth four times that of the control. These results indicate that AA-2G is gradually cleaved by the cellular alpha-glucosidase to release L-ascorbic acid, which adequately stimulates collagen synthesis and proliferation of human skin fibroblasts. PMID:1552361

Yamamoto, I; Muto, N; Murakami, K; Akiyama, J

1992-04-01

163

Gene expression profiling in wild-type and metallothionein mutant fibroblast cell lines  

Directory of Open Access Journals (Sweden)

Full Text Available The role of metallothioneins (MT in copper homeostasis is of great interest, as it appears to be partially responsible for the regulation of intracellular copper levels during adaptation to extracellular excess of the metal. To further investigate a possible role of MTs in copper metabolism, a genomics approach was utilized to evaluate the role of MT on gene expression. Microarray analysis was used to examine the effects of copper overload in fibroblast cells from normal and MT I and II double knock-out mice (MT-/-. As a first step, we compared genes that were significantly upregulated in wild-type and MT-/- cells exposed to copper. Even though wild-type and mutant cells are undistinguishable in terms of their morphological features and rates of growth, our results show that MT-/- cells do not respond with induction of typical markers of cellular stress under copper excess conditions, as observed in the wild-type cell line, suggesting that the transcription initiation rate or the mRNA stability of stress genes is affected when there is an alteration in the copper store capacity. The functional classification of other up-regulated genes in both cell lines indicates that a large proportion (>80% belong to two major categories: 1 metabolism; and 2 cellular physiological processes, suggesting that at the transcriptional level copper overload induces the expression of genes associated with diverse molecular functions. These results open the possibility to understand how copper homeostasis is being coordinated with other metabolic pathways.

ÁNGELA D ARMENDÁRIZ

2006-01-01

164

Activation of NF-?B in human skin fibroblasts by the oxidative stress generated by UVA radiation  

International Nuclear Information System (INIS)

We have examined the role of the nucleus and the membrane in the activation of nuclear factor (NF)-kB by oxidant stress generated via the UVA (320-380 nm) component of solar radiation. Nuclear extracts from human skin fibroblasts that had been irradiated with UVA at doses that caused little DNA damage contained activated NF-?B that bound to its recognition sequence in DNA. The UVA radiation-dependent activation of NF-?B in enucleated cells confirmed that the nucleus was not involved. On the other hand, UVA radiation-dependent activation of NF-?B appeared to be correlated with membrane damage, and activation could be prevented by ?-tocopherol and butylated hydroxytoluene, agents that inhibited UVA radiation-dependent peroxidation of cell membrane lipids. The activation of NF-?B by the DNA damaging agents UVC (200-290 nm) and UVB (290-320 nm) radiation also only occurred at doses where significant membrane damage was induced, and, overall, activation was not correlated with the relative levels of DNA damage induced by UVC/UVB and UVA radiations. We conclude that the oxidative modification of membrane components may be an important factor to consider in the UV radiation-dependent activation of NF-?B over all wavelength ranges examined. (Author)

1995-09-01

165

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

Energy Technology Data Exchange (ETDEWEB)

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 mechanism.

Haddock, R.C.; Mack, P.; Leal, S.; Baenziger, N.L. (Washington Univ. School of Medicine, St. Louis, MO (USA))

1990-08-25

166

Activation of NF-{kappa}B in human skin fibroblasts by the oxidative stress generated by UVA radiation  

Energy Technology Data Exchange (ETDEWEB)

We have examined the role of the nucleus and the membrane in the activation of nuclear factor (NF)-kB by oxidant stress generated via the UVA (320-380 nm) component of solar radiation. Nuclear extracts from human skin fibroblasts that had been irradiated with UVA at doses that caused little DNA damage contained activated NF-{kappa}B that bound to its recognition sequence in DNA. The UVA radiation-dependent activation of NF-{kappa}B in enucleated cells confirmed that the nucleus was not involved. On the other hand, UVA radiation-dependent activation of NF-{kappa}B appeared to be correlated with membrane damage, and activation could be prevented by {alpha}-tocopherol and butylated hydroxytoluene, agents that inhibited UVA radiation-dependent peroxidation of cell membrane lipids. The activation of NF-{kappa}B by the DNA damaging agents UVC (200-290 nm) and UVB (290-320 nm) radiation also only occurred at doses where significant membrane damage was induced, and, overall, activation was not correlated with the relative levels of DNA damage induced by UVC/UVB and UVA radiations. We conclude that the oxidative modification of membrane components may be an important factor to consider in the UV radiation-dependent activation of NF-{kappa}B over all wavelength ranges examined. (Author).

Vile, G.F.; Tanew-Iliitschew, Adrian; Tyrrell, R.M. [Institut Suisse de Recherches Experimentales sur le Cancer, Lausanne (Switzerland)

1995-09-01

167

Peroxisomal very long-chain fatty acid [beta]-oxidation in human skin fibroblasts: activity in Zellweger syndrome and other peroxisomal disorders  

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Since very long-chain fatty acids with a chain length of 24 carbons or more are known to accumulate in tissues and body fluids from patients with the cerebro-hepato-renal (Zellweger) syndrome, infantile Refsum disease, neonatal adrenoleukodystrophy and X-linked adrenoleukodystrophy, we studied very long-chain fatty acid oxidation in cultured skin fibroblasts from these patients. In this paper, we report that in accordance with earlier results the first step in the ?-oxidation of the very lon...

Wanders, R. J. A.; Roermund, C. W. T.; Wijland, M. J. A.; Heikoop, J.; Schutgens, R. B. H.; Schram, A. W.; Tager, J. M.; Bosch, H. Den; Poll-the?, B. T.; Saudubray, J. M.; Moser, H. W.; Moser, A. B.

1987-01-01

168

Demonstration of elastin gene expression in human skin fibroblast cultures and reduced tropoelastin production by cells from a patient with atrophoderma.  

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Atrophoderma is a rare dermal disorder characterized by a patchy distribution of areas apparently devoid of elastic fibers. Skin fibroblast cultures were established from the normal and affected dermis of a patient with this disorder. Human tropoelastin was identified in culture medium by use of electroblotting and anti-elastin antisera. An enzyme-linked immunosorbent assay was used to establish that significantly less elastin accumulated in the media of cultured cells from lesional fibroblas...

Giro, M. G.; Oikarinen, A. I.; Oikarinen, H.; Sephel, G.; Uitto, J.; Davidson, J. M.

1985-01-01

169

Enhancement human cytomegalovirus replication in a human lung fibroblast cell line by interleukin-8.  

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We examined the effects of interleukin-8 (IL-8) on cytomegalovirus (CMV) replication in human fibroblasts. Exposure of fibroblasts to IL-8 augmented both infectious virus production and replication of CMV, with concomitant increases in the levels of both the transcript of the CMV pp71 genome and the synthesis of the CMV late antigen. We also found that CMV selectively induced transcripts of the IL-8 type 1 receptor in fibroblasts. These results suggest that IL-8 also contributes to inflammato...

Murayama, T.; Kuno, K.; Jisaki, F.; Obuchi, M.; Sakamuro, D.; Furukawa, T.; Mukaida, N.; Matsushima, K.

1994-01-01

170

Altered binding of /sup 125/I-labeled calmodulin to a 46. 5-kilodalton protein in skin fibroblasts cultured from patients with cystic fibrosis  

Energy Technology Data Exchange (ETDEWEB)

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 Ca/sup 2 +//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 /sup 125/I-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.

Tallant, E.A.; Wallace, R.W.

1987-02-01

171

Altered binding of "1"2"5I-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 Ca"2"+/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 "1"2"5I-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

1987-01-01

172

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

173

Irradiation effect on the apoptosis induction in the human cancer cell lines and the gingival fibroblast  

International Nuclear Information System (INIS)

The radiation-induced apoptosis was studied for two human cancer cell lines (KB cells, RPMI 2650 cells) and the human gingival fibroblast cell line (HGF-1 cells). The single irradiation of 2, 10, 20 Gy was done with 241.5 cGy/min dose rate using the 137 Cs MK cell irradiator. The cell were stained with propidium iodide and examined under the fluoro-microscope and assayed with the flow cytometry a day after irradiation. Also, the LDH assay was done to determine the amount of necrotic cells. The obtained results were as follows : 1. On the fluoro-microscope, many fragmented nuclei were detected in the KB, RPMI 2650, and HGF-1 cells after irradiation. 2. On the DNA content histogram obtained from the flow cytometry, the percentages of the pre-G1 peak of the control and 2, 10 and 20 Gy irradiation group were 4.5, 55.0, 52.3, and 66.6% on KB cells, 2.7, 3.3, 31.8, and 32.6% on RPMI 2650 cells and 2.8, 21.8, 30.4, and 40.2% on HGF-1 cells respectively. 3. The number of G1-stage cells was abruptly decreased after 2 Gy irradiation on KB cells and 10 Gy irradiation on RPMI 2650 cells, But there was a slight decrease without regard to irradiation dose on HGF-1 cells. 4. There was no significantly different absorbance in extracellular LDH assay along the experimental cell lines

1998-02-01

174

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

175

The effect of xanthorrhizol on the expression of matrix metalloproteinase-1 and type-I procollagen in ultraviolet-irradiated human skin fibroblasts.  

Science.gov (United States)

Matrix metalloproteinases (MMPs) are key regulators of the skin photoaging process that is set in motion by exposure to ultraviolet (UV) irradiation. This skin damage results from UV-induced generation of reactive oxygen species, which are associated with upregulation of MMPs and decreased collagen synthesis. We investigated the effects of xanthorrhizol, isolated from Curcuma xanthorrhiza, on the expression of MMP-1 and type-I procollagen in UV-irradiated human skin fibroblasts. Fibroblasts cultured in the presence or absence of purified xanthorrhizol or C. xanthorrhiza extract were irradiated with UV (20 mJ/cm(2)), and MMP-1 and type-I procollagen levels were measured using Western blot analysis. Xanthorrhizol (0.001-0.1 microM) and C. xanthorrhiza extract (0.01-0.5 microg/mL) induced a significant, dose-dependent decrease in the expression of MMP-1 protein, and increased the expression of type-1 procollagen. At a concentration of 0.1 microM, xanthorrhizol nearly completely abrogated MMP-1 expression. The MMP-1-suppressing and type-1 procollagen-inducing effects of xanthorrhizol treatment were greater than those of epigallocatechin 3-O-gallate (EGCG), which is known to be a natural anti-aging agent. These results suggest that xanthorrhizol is a potential candidate for the prevention and treatment of skin aging. PMID:19277961

Oh, Hyun-In; Shim, Jae-Seok; Gwon, Song-Hui; Kwon, Ho-Jeong; Hwang, Jae-Kwan

2009-09-01

176

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

177

Gene expression profiling in wild-type and metallothionein mutant fibroblast cell lines  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in english The role of metallothioneins (MT) in copper homeostasis is of great interest, as it appears to be partially responsible for the regulation of intracellular copper levels during adaptation to extracellular excess of the metal. To further investigate a possible role of MTs in copper metabolism, a geno [...] mics approach was utilized to evaluate the role of MT on gene expression. Microarray analysis was used to examine the effects of copper overload in fibroblast cells from normal and MT I and II double knock-out mice (MT-/-). As a first step, we compared genes that were significantly upregulated in wild-type and MT-/- cells exposed to copper. Even though wild-type and mutant cells are undistinguishable in terms of their morphological features and rates of growth, our results show that MT-/- cells do not respond with induction of typical markers of cellular stress under copper excess conditions, as observed in the wild-type cell line, suggesting that the transcription initiation rate or the mRNA stability of stress genes is affected when there is an alteration in the copper store capacity. The functional classification of other up-regulated genes in both cell lines indicates that a large proportion (>80%) belong to two major categories: 1) metabolism; and 2) cellular physiological processes, suggesting that at the transcriptional level copper overload induces the expression of genes associated with diverse molecular functions. These results open the possibility to understand how copper homeostasis is being coordinated with other metabolic pathways.

ÁNGELA D, ARMENDÁRIZ; FELIPE, OLIVARES; RODRIGO, PULGAR; ALEX, LOGUINOV; VERÓNICA, CAMBIAZO; CHRISTOPHER D, VULPE; MAURICIO, GONZÁLEZ.

178

Transformation of cultured human embryonic fibroblasts by oncornavirus-like particles released from a human carcinoma cell line.  

Science.gov (United States)

A fibroblast-like cell culture was established from a stomach biopsy of a patient with metastatic adenocarcinoma. One of the cultures, at the 6th passage level, left unattended for a month at 37 degrees, produced numerous foci of epithelioid cells. Upon subculturing, an epithelioid cell line, designated HCCL (human carcinoma cell line), was established. The HCCL cells released particles possessing the characteristics of oncornaviruses: density 1.175 g/ml, cores with a density of 1.22-1.26 g/ml, high-molecular-weight RNA (60-70S) and RNA-instructed DNA polymerase activity (deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 2.7.7.7). Inoculation of particles released from HCCL cells into cultures of human embryo muscle fibroblasts resulted in the appearance of foci of transformed cells. PMID:52157

Balabanova, H; Kotler, M; Becker, Y

1975-07-01

179

Host-cell reactivation of uv-irradiated and chemically treated Herpes simplex virus type 1 strain MP in normal and xeroderma pigmentosum skin fibroblasts  

International Nuclear Information System (INIS)

The host-cell reactivation of UV-irradiated and N-acetoxy-2-acetylaminofluorene-treated herpes simplex virus type 1 strain mp was studied in normal human skin fibroblasts and xeroderma pigmentosum skin fibroblasts from XP genetic complementation groups A-D and in an XP variant. The increasing relative order for the host-cell reactivation of both types of damaged virus in the different complementation groups is A = D < B < C; XP variant = normal controls. XP complementation group D cells, which manifest the most severe inhibition of her ability for both UV-irradiated and N-acetoxy-2-acetylaminofluorene-treated virus, can reactivate nitrogen mustard treated HSV-1 mp to the same extent as normal cells. Together, these results indicate that (1) Excision repair of UV and N-acetoxy-2-acetylaminofluorene DNA damaged viruses share a common rate limiting enzymatic step and (2) The repair defect in xeroderma pigmentosum cells plays little or no role in the recovery of nitrogen mustard treated virus. The results of studies on the effect of caffeine on the survival of both UV- and N-acetoxy-2-acetylaminofluorene-treated virus in normal and XP cells imply that the reactivation of HSV-1 mp is mediated by an excision repair process with little if any recovery contributed by post-replication repair mechanisms. The host-cell reactivation of N-acetoxy-2-acetylaminofluorene-treated HSV-1 mp was also correlated with the defective UV-induced unscheduled DNA synthesis in two skin fibroblast strains established from a skin biopsy obtained from each of two juvenile females who had been clinically diagnosed as xeroderma pigmentosum. These findings are discussed in relation to the further characterization of the xeroderma pigmentosum phenotype and their possible utilization for the selection and isolation of new mammalian cell DNA repair mutants

1976-01-01

180

Differential response of human adipose tissue-derived mesenchymal stem cells, dermal fibroblasts, and keratinocytes to burn wound exudates: potential role of skin-specific chemokine CCL27.  

Science.gov (United States)

Many cell-based regenerative medicine strategies toward tissue-engineered constructs are currently being explored. Cell-cell interactions and interactions with different biomaterials are extensively investigated, whereas very few studies address how cultured cells will interact with soluble wound-healing mediators that are present within the wound bed after transplantation. The aim of this study was to determine how adipose tissue-derived mesenchymal stem cells (ASC), dermal fibroblasts, and keratinocytes will react when they come in contact with the deep cutaneous burn wound bed. Burn wound exudates isolated from deep burn wounds were found to contain many cytokines, including chemokines and growth factors related to inflammation and wound healing. Seventeen mediators were identified by ELISA (concentration range 0.0006-9 ng/mg total protein), including the skin-specific chemokine CCL27. Burn wound exudates activated both ASC and dermal fibroblasts, but not keratinocytes, to increase secretion of CXCL1, CXCL8, CCL2, and CCL20. Notably, ASC but not fibroblasts or keratinocytes showed significant increased secretion of vascular endothelial growth factor (5-fold) and interleukin-6 (253-fold), although when the cells were incorporated in bi-layered skin substitute (SS) these differences were less pronounced. A similar discrepancy between ASC and dermal fibroblast mono-cultures was observed when recombinant human-CCL27 was used instead of burn wound exudates. Although CCL27 did not stimulate the secretion of any of the wound-healing mediators by keratinocytes, these cells, in contrast to ASC or dermal fibroblasts, showed increased proliferation and migration. Taken together, these results indicate that on transplantation, keratinocytes are primarily activated to promote wound closure. In contrast, dermal fibroblasts and, in particular, ASC respond vigorously to factors present in the wound bed, leading to increased secretion of angiogenesis/granulation tissue formation factors. Our findings have implications for the choice of cell type (ASC or dermal fibroblast) to be used in regenerative medicine strategies and indicate the importance of taking into account interactions with the wound bed when developing advanced therapies for difficult-to-close cutaneous wounds. PMID:23980822

van den Broek, Lenie J; Kroeze, Kim L; Waaijman, Taco; Breetveld, Melanie; Sampat-Sardjoepersad, Shakun C; Niessen, Frank B; Middelkoop, Esther; Scheper, Rik J; Gibbs, Susan

2014-01-01

 
 
 
 
181

Enhanced in vitro radiosensitivity of skin fibroblasts in two patients developing brain necrosis following AVM radiosurgery: a new risk factor with potential for a predictive assay  

International Nuclear Information System (INIS)

Purpose: Radiosurgery is an effective treatment for arteriovenous malformations (AVM) with a low risk of developing brain necrosis. Models have been developed to predict the risk of complications. We postulated that genetic differences in radiosensitivity may also be a risk factor. Methods and Materials: Fibroblast cultures were established from skin biopsies in two AVM patients developing radiation necrosis. The results of clonogenic survival assays were compared to a parallel study with two groups of cancer patients treated with radiation: 1) patients without late side effects; 2) patients experiencing severe late sequelae. Results: The survival fraction at 2 Gy (SF2) of the 2 AVM patients was 0.17 (0.14-0.19) and 0.18 (0.14-0.22). The SF2's of the cancer patients ranged between 0.25-0.38 (mean = 0.31) for the control group, and between 0.10-0.20 (mean = 0.17) for the hypersensitive group. The SF2's of the AVM patients who developed brain necrosis were comparable to that of the hypersensitive group (p = 0.85) but significantly lower than the control group (p = 0.05). Conclusion: The two patients who developed radiation necrosis demonstrate increased fibroblast radiosensitivity. The SF2 of skin fibroblasts may potentially be used as a predictive assay to detect patients at risk for brain necrosis

2000-04-01

182

Correlation between normal tissue complications and in vitro radiosensitivity of skin fibroblasts derived from radiotherapy patients treated for variety of tumors  

International Nuclear Information System (INIS)

Purpose: To assess the relationship between fibroblast intrinsic radiosensitivity in vitro and late reactions of normal tissues in patients treated by definitive radiotherapy for variety of tumors. Patients and Methods: Ten patients were selected for this study. They were treated by radical radiotherapy for variety of tumors, including non-Hodgkin's lymphoma, prostate, glottic larynx, anal canal, cervix, bladder, thyroid gland, and tonsil pillar. Five patients did not develop any significant late reactions (normally sensitive group, NS). The other five developed late complications in different normal tissues and organs that proved to be fatal in one patient (clinically hyper-sensitive group, HS). Fibroblast cultures were established from punch skin biopsy and radiosensitivity in vitro was measured. The survival fraction at 2 Gy (SF2) was calculated and compared between the two groups. Results: SF2 ranged between 0.10 and 0.38 with a mean of 0.24. The mean SF2 for each of the NS and the HS groups were 0.31 and 0.17, respectively. The non-parametric rank test of Mann-Whitney shows that the difference between the two groups is statistically significant (p = 0.01). Conclusion: This study indicates that the in vitro radiosensitivity of skin fibroblasts is correlated with late complications in different organs and normal tissues following radiotherapy for variety of tumors. It also lends support to the existence of a common genetic component determining the radiosensitivity of cells targeted by the late effects of ionizing radiation. Key words:

2000-01-01

183

Absence of correlations between the radiosensitivity of human T-lymphocytes at G_0 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 D_1_0 (the dose required to kill 90 % of the cells) ±SD was 3.58 ± 0.21 Gy for T-lymphocytes irradiated at G_0 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 D_1_0 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)

1990-01-01

184

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

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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

185

PMN Leukocytes and Fibroblasts Numbers on Wound Burn Healing on the Skin of White Rat after Administration of Ambonese Plantain Banana  

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Full Text Available A study of ambonese plantain banana (Musa paradisiaca var sapientum Lamb treatment in burn wound healing on the skin of white rats (Rattus novergicus has been conducted. The wound healing of burn injuries was evaluated by counting the number of PMN leukocytes and fibroblasts at the 7th, 14th, and 21st days following the treatment. The study showed that the decrease in number of PMN leukocytes of subjects treated with ambonese plantain banana was relatively more significant compared to both negative and positive control (Bioplacenton ®. In contrast, an increasing number of fibroblasts was significantly demonstrated at the 14th and 21st days after treatment. In conclusion, ambonese plantain banana treatment in burn injuries will provide better results compared to both positive and negative controls.

Juniarti

2012-04-01

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.  

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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

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  

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)

2012-10-01

188

Expression of two different forms of fibroblast growth factor receptor 1 in different mouse tissues and cell lines.  

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The fibroblast growth factor receptors (FGFRs) form a multigene family of at least four members, all having extracellular regions consisting of either two or three immunoglobin-like (Ig-like) domains. By RNase protection analysis we have analyzed the expression of FGFR-1 mRNA in various tissues and cell lines and demonstrated that all of the cell lines studied expressed at least two different forms of the FGFR-1 at similar levels. Although muscle and heart express forms having either two [FGF...

Bernard, O.; Li, M.; Reid, H. H.

1991-01-01

189

Protection against TGF-?1-induced fibrosis effects of IL-10 on dermal fibroblasts and its potential therapeutics for the reduction of skin scarring.  

Science.gov (United States)

Scarring, tightly associated with fibrosis, is a significant symptomatic clinical problem. Interleukin 10 (IL-10) has been identified as a candidate scar-improving therapy based on preclinical studies. However, the molecular mechanism of IL-10 in scar improvement is still uncertain. In this study, human dermal fibroblasts stimulated with TGF-?1 were treated with IL-10 to analyze the mRNA and some of proteins' expression levels of type I collagen (Col1), type III collagen (Col3), alpha-smooth muscle actin (?-SMA), matrix metalloproteinase-1 (MMP1), MMP2, MMP8 and tissue inhibitor of metalloproteinase 1 (TIMP1), TIMP2 by real-time PCR and Western blot, to observe ?-SMA-positive fibroblasts by immunocytochemistry. The contracture and improvement of fibroblast-populated collagen lattice (FPCL) and a murine model of wound healing were used to evaluate the scar-improving effects by histological staining. The results showed that IL-10 can significantly down-regulate the mRNA and protein expression levels of Col1, Col3, ?-SMA, and up-regulate the mRNA expression levels of MMP1 and MMP8, and decrease ?-SMA-positive fibroblasts. FPCL analysis showed that the IL-10 (20 ng/ml) can significantly inhibit the contracture, improve the architecture of FPCL. Wounds injected with IL-10 demonstrated that the appearance of scar was improved, the wound margin of scarring was narrow, and the deposition of collagens (Col1 and Col3) in regenerated tissue was relieved. These results provide direct evidences that IL-10 has the inhibitory effects on the excessive deposition of extracellular matrix components and fibroblast-to-myofibroblast transition, and show that IL-10 has the potential therapy in prevention and reduction of skin scarring. PMID:23321694

Shi, Ji-Hong; Guan, Hao; Shi, Shan; Cai, Wei-Xia; Bai, Xiao-Zhi; Hu, Xiao-Long; Fang, Xiao-Bin; Liu, Jia-Qi; Tao, Ke; Zhu, Xiong-Xiang; Tang, Chao-Wu; Hu, Da-Hai

2013-05-01

190

Relaxin regulates fibrillin 2, but not fibrillin 1, mRNA and protein expression by human dermal fibroblasts and murine fetal skin.  

Science.gov (United States)

Relaxin modulates connective tissue remodeling by altering matrix molecule expression. We have found that relaxin specifically inhibits a microfibril component, fibrillin 2 (FBN2), without affecting fibrillin 1 (FBN1). Human dermal fibroblasts (HDFs) grown or stimulated to overexpress fibrillin expression were used to show that relaxin specifically down-regulated FBN2 mRNA and protein levels. Continuous exposure of HDFs to relaxin (30ng/ml) significantly (PFBN2 mRNA and protein in fetal skin from pregnant relaxin knockout mice. The regulation of FBN2 expression may be associated with functional changes in elastic tissues during development and growth. PMID:12590922

Samuel, Chrishan S; Sakai, Lynn Y; Amento, Edward P

2003-03-01

191

Fibroblast growth inhibitor.  

Science.gov (United States)

A new cytokine has been recognized in the conditioned media (CM) of freshly isolated acute myelocytic leukemia cells, cultured with 12-0-tetradecanayl phorbol acetate (TPA) 10(-8)M. The fraction with 70,000 MW was separated from CM by ammonium sulfate precipitation, ion-exchange cation and anion chromatography, and Sephadex G-200 gel filtration. It was a fibroblast growth inhibitor (FGI). This substance stopped fetal and skin (MALME 3 line) fibroblast propagation. The cytostatic effect was reversible on removal of FGI. At the same time, FGI did not inhibit macrophage proliferation. The fraction stimulated formation of monocytic and granulocytic colonies altered the phenotype of human U-2 osteosarcoma cells grown from epithelial-like to fibroblast-like cells, and stimulated differentiation of leukemic cells along the macrophage path. Some cells of promyelocytic leukemia line HL-60, grown in the presence of FGI, were stimulated to differentiate and some underwent lysis. The response to FGI of cells from different patients varied. PMID:3224159

Rogalsky, V; Svet-Moldavsky, I; Arlin, Z; Den, T; Holland, J F

1988-01-01

192

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

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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

193

Deciphering the differential response of two human fibroblast cell lines following Chikungunya virus infection  

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Abstract Background Chikungunya virus (CHIKV) is an arthritogenic member of the Alphavirus genus (family Togaviridae) transmitted by Aedes mosquitoes. CHIKV is now known to target non hematopoietic cells such as epithelial, endothelial cells, fibroblasts and to less extent monocytes/macrophages. The type I interferon (IFN) response is an early innate immune mechanism that protects cells against viral infection. Cells express different pattern recogn...

Thon-Hon Vincent G; Denizot Melanie; Li-Pat-Yuen Ghislaine; Giry Claude; Jaffar-Bandjee Marie-Christine; Gasque Philippe

2012-01-01

194

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)

2002-06-01

195

Studies of DNA and chromosome damage in skin fibroblasts and blood lymphocytes from psoriasis patients treated with 8-methoxypsoralen and UVA irradiation  

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Exposure of human lymphocytes and skin fibroblasts in vitro to a single, clinically used dose of PUVA, i.e., 0.1 micrograms/ml of 8-methoxypsoralen (8-MOP) plus 0.9-4 J/cm2 of longwave ultraviolet radiation (UVA), lead to the formation of DNA damage as determined by alkaline elution, and to chromosome aberrations and sister chromatid exchanges (SCE). When lymphocyte-enriched plasma was obtained from psoriasis patients 2 h after oral intake of 8-MOP and then UVA irradiated (1.8-3.6 J/cm2) in vitro, an increased frequency of chromosome aberrations and SCE was observed. Normal levels of chromosome aberrations and SCE were found in lymphocytes of psoriasis patients after 3-30 weeks of PUVA treatment in vivo. A small but statistically significant increase in the SCE frequency was observed in the lymphocytes of psoriasis patients treated for 1-6 years with PUVA (mean 18.0 SCE/cell) as compared with before PUVA (mean 15.8, p less than 0.05). Skin fibroblasts of psoriasis patients analyzed 5 years after the start of PUVA treatment showed a normal number of SCE but a high fraction of filter-retained DNA in the alkaline elution assay, suggesting the presence of cross-linked DNA.

Bredberg, A.; Lambert, B.; Lindblad, A.; Swanbeck, G.; Wennersten, G.

1983-08-01

196

Studies of DNA and chromosome damage in skin fibroblasts and blood lymphocytes from psoriasis patients treated with 8-methoxypsoralen and UVA irradiation  

International Nuclear Information System (INIS)

Exposure of human lymphocytes and skin fibroblasts in vitro to a single, clinically used dose of PUVA, i.e., 0.1 micrograms/ml of 8-methoxypsoralen (8-MOP) plus 0.9-4 J/cm2 of longwave ultraviolet radiation (UVA), lead to the formation of DNA damage as determined by alkaline elution, and to chromosome aberrations and sister chromatid exchanges (SCE). When lymphocyte-enriched plasma was obtained from psoriasis patients 2 h after oral intake of 8-MOP and then UVA irradiated (1.8-3.6 J/cm2) in vitro, an increased frequency of chromosome aberrations and SCE was observed. Normal levels of chromosome aberrations and SCE were found in lymphocytes of psoriasis patients after 3-30 weeks of PUVA treatment in vivo. A small but statistically significant increase in the SCE frequency was observed in the lymphocytes of psoriasis patients treated for 1-6 years with PUVA (mean 18.0 SCE/cell) as compared with before PUVA (mean 15.8, p less than 0.05). Skin fibroblasts of psoriasis patients analyzed 5 years after the start of PUVA treatment showed a normal number of SCE but a high fraction of filter-retained DNA in the alkaline elution assay, suggesting the presence of cross-linked DNA

1983-01-01

197

Recombinant growth factor mixtures induce cell cycle progression and the upregulation of type I collagen in human skin fibroblasts, resulting in the acceleration of wound healing processess.  

Science.gov (United States)

Application of growth factor mixtures has been used for wound healing and anti-wrinkles agents. The aim of this study was to evaluate the effect of recombinant growth factor mixtures (RGFM) on the expression of cell cycle regulatory proteins, type I collagen, and wound healing processes of acute animal wound models. The results showed that RGFM induced increased rates of cell proliferation and cell migration of human skin fibroblasts (HSF). In addition, expression of cyclin D1, cyclin E, cyclin-dependent kinase (Cdk)4, and Cdk2 proteins was markedly increased with a growth factor mixtures treatment in fibroblasts. Expression of type I collagen was also increased in growth factor mixtures-treated HSF. Moreover, growth factor mixtures-induced the upregulation of type I collagen was associated with the activation of Smad2/3. In the animal model, RGFM-treated mice showed accelerated wound closure, with the closure rate increasing as early as on day 7, as well as re-epithelization and reduced inflammatory cell infiltration than phosphate-buffered saline (PBS)-treated mice. In conclusion, the results indicated that RGFM has the potential to accelerate wound healing through the upregulation of type I collagen, which is partly mediated by activation of Smad2/3-dependent signaling pathway as well as cell cycle progression in HSF. The topical application of growth factor mixtures to acute and chronic skin wound may accelerate the epithelization process through these molecular mechanisms. PMID:24626875

Lee, Do Hyun; Choi, Kyung-Ha; Cho, Jae-We; Kim, So Young; Kwon, Tae Rin; Choi, Sun Young; Choi, Yoo Mi; Lee, Jay; Yoon, Ho Sang; Kim, Beom Joon

2014-05-01

198

Tamarind seed coat extract restores reactive oxygen species through attenuation of glutathione level and antioxidant enzyme expression in human skin fibroblasts in response to oxidative stress  

Science.gov (United States)

Objective To investigate the role and mechanism of tamarind seed coat extract (TSCE) on normal human skin fibroblast CCD-1064Sk cells under normal and oxidative stress conditions induced by hydrogen peroxide (H2O2). Methods Tamarind seed coats were extracted with boiling water and then partitioned with ethyl acetate before the cell analysis. Effect of TSCE on intracellular reactive oxygen species (ROS), glutathione (GSH) level, antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase activity including antioxidant protein expression was investigated. Results TSCE significantly attenuated intracellular ROS in the absence and presence of H2O2 by increasing GSH level. In the absence of H2O2, TSCE significantly enhanced SOD and catalase activity but did not affected on GPx. Meanwhile, TSCE significantly increased the protein expression of SOD and GPx in H2O2-treated cells. Conclusions TSCE exhibited antioxidant activities by scavenging ROS, attenuating GSH level that could protect human skin fibroblast cells from oxidative stress. Our results highlight the antioxidant mechanism of tamarind seed coat through an antioxidant enzyme system, the extract potentially benefits for health food and cosmeceutical application of tamarind seed coat.

Nakchat, Oranuch; Nalinratana, Nonthaneth; Meksuriyen, Duangdeun; Pongsamart, Sunanta

2014-01-01

199

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

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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

200

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.)

1981-01-01

 
 
 
 
201

Skin fibroblasts from pantothenate kinase-associated neurodegeneration patients show altered cellular oxidative status and have defective iron-handling properties.  

Science.gov (United States)

Pantothenate kinase-associated neurodegeneration (PKAN) is a neurodegenerative disease belonging to the group of neurodegeneration with brain iron accumulation disorders. It is characterized by progressive impairments in movement, speech and cognition. The disease is inherited in a recessive manner due to mutations in the Pantothenate Kinase-2 (PANK2) gene that encodes a mitochondrial protein involved in Coenzyme A synthesis. To investigate the link between a PANK2 gene defect and iron accumulation, we analyzed primary skin fibroblasts from three PKAN patients and three unaffected subjects. The oxidative status of the cells and their ability to respond to iron were analyzed in both basal and iron supplementation conditions. In basal conditions, PKAN fibroblasts show an increase in carbonylated proteins and altered expression of antioxidant enzymes with respect to the controls. After iron supplementation, the PKAN fibroblasts had a defective response to the additional iron. Under these conditions, ferritins were up-regulated and Transferrin Receptor 1 (TfR1) was down-regulated to a minor extent in patients compared with the controls. Analysis of iron regulatory proteins (IRPs) reveals that, with respect to the controls, PKAN fibroblasts have a reduced amount of membrane-associated mRNA-bound IRP1, which responds imperfectly to iron. This accounts for the defective expression of ferritin and TfR1 in patients' cells. The inaccurate quantity of these proteins produced a higher bioactive labile iron pool and consequently increased iron-dependent reactive oxygen species formation. Our results suggest that Pank2 deficiency promotes an increased oxidative status that is further enhanced by the addition of iron, potentially causing damage in cells. PMID:22692681

Campanella, Alessandro; Privitera, Daniela; Guaraldo, Michela; Rovelli, Elisabetta; Barzaghi, Chiara; Garavaglia, Barbara; Santambrogio, Paolo; Cozzi, Anna; Levi, Sonia

2012-09-15

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

Inhibitory Effects of Terminalia catappa on UVB-Induced Photodamage in Fibroblast Cell Line  

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This study investigated whether Terminalia catappa L. hydrophilic extract (TCLW) prevents photoaging in human dermal fibroblasts after exposure to UVB radiation. TCLW exhibited DPPH free radical scavenging activity and protected erythrocytes against AAPH-induced hemolysis. In the gelatin digestion assay, the rates of collagenase inhibition by TCL methanol extract, TCLW, and its hydrolysates were greater than 100% at the concentration of 1?mg/mL. We found that serial dilutions of TCLW (10–...

Kuo-Ching Wen; I-Chen Shih; Jhe-Cyuan Hu; Sue-Tsai Liao; Tsung-Wei Su; Hsiu-Mei Chiang

2011-01-01

204

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

205

Skin tension lines and preferable incisions in context of biomechanical skin characteristics. Linie napi?cia i wyboru kierunków naci?? skóry w kontek?cie jej biomechanicznych w?a?ciwo?ci.  

Directory of Open Access Journals (Sweden)

Full Text Available The proper orientation of skin incisions influence the functional and aesthetic results in dermatosurgery. Langer’s lines, dynamic wrinkle expression lines and relaxed skin tension lines are the basic marks for the planning of incisions. Fundamental biomechanical skin properies, such as skin elasticity, strain, creep and stress relaxation are disscused. The knowledge of these properties contributes to effective planning and execution of surgical skin procesures.

Adam W?odarkiewicz

2009-09-01

206

Adaptive response to ionizing radiation in normal human skin fibroblasts. Enhancement of DNA repair rate and modulation of gene expression  

International Nuclear Information System (INIS)

Low doses and dose rates of ionizing radiation enhance the rate of DNA repair in human fibroblasts and protect the cells against radiation-induced micronucleus formation. Chronic exposures reduce the mRNA levels of the genes topoisomerase II and FACC-1 (Fanconi's anemia, group C). (authors). 11 refs., 1 tab., 2 figs

1994-01-01

207

Collagen expression in fibroblasts with a novel LMNA mutation  

International Nuclear Information System (INIS)

Laminopathies are a group of genetic disorders caused by LMNA mutations; they include muscular dystrophies, lipodystrophies, and progeroid syndromes. We identified a novel heterozygous LMNA mutation, L59R, in a patient with the general appearance of mandibuloacral dysplasia and progeroid features. Examination of the nuclei of dermal fibroblasts revealed the irregular morphology characteristic of LMNA mutant cells. The nuclear morphological abnormalities of LMNA mutant lymphoblastoid cell lines were less prominent compared to those of primary fibroblasts. Since it has been reported that progeroid features are associated with increased extracellular matrix in dermal tissues, we compared a subset of these components in fibroblast cultures from LMNA mutants with those of control fibroblasts. There was no evidence of intracellular accumulation or altered mobility of collagen chains, or altered conversion of procollagen to collagen, suggesting that skin fibroblast-mediated matrix production may not play a significant role in the pathogenesis of this particular laminopathy

2007-01-19

208

The wavelength dependence of u.v.-induced pyrimidine dimer formation, cell killing and mutation induction in human diploid skin fibroblasts  

International Nuclear Information System (INIS)

We determined the wavelength dependence of u.v.-induced pyrimidine dimer formation, cell killing and mutation induction in human diploid skin fibroblasts. Pyrimidine dimers were quantified using the T4 endonuclease V assay, cell killing was measured as loss of colony forming ability and mutation induction was detected at the HPRT locus. U.v. irradiation was performed with monochromatic light of four different wavelengths (254, 297, 302 and 365 nm) and with polychromatic light of a Philips TL-01 lamp (predominantly 312 nm). The relative wavelength dependence for cell killing and mutation induction did not correlate with that for dimer formation. Toxicity and mutagenicity per equivalent initial dimer load increase with increasing wavelength. The relative wavelength dependence for cell killing and mutation induction is essentially the same, except at 365 nm. (author)

1986-01-01

209

Induction of the heme oxygenase gene in human skin fibroblasts by hydrogen peroxide and UVA (365 nm) radiation: evidence for the involvement of the hydroxyl radical  

Energy Technology Data Exchange (ETDEWEB)

The induction of heme oxygenase by both hydrogen peroxide and UVA (365 nm) radiation in normal human skin fibroblasts is prevented by prior treatment of cells with the specific iron chelators, o-phenanthroline or desferrioxamine. In addition, both iron chelators protected cells against the lethal effects of H{sub 2}O{sub 2} treatment or UVA irradiation. We propose that the generation of the highly reactive hydroxyl radical by an iron catalyzed Fenton reaction is involved both in the induction of this stress response and, at least in part, in cell killing by the two treatments. These results are also consistent with the idea that the heme oxygenase gene is induced in response to oxidative stress and that its induction may constitute an inducible protective mechanism against oxidative damage induced by both hydrogen peroxide and UVA radiation. (author).

Keyse, S.M.; Tyrrell, R.M. (Swiss Inst. for Experimental Cancer Research, Epalinges (Switzerland))

1990-05-01

210

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

211

Pentoxifylline modifies three-dimensional collagen lattice model contraction and expression of collagen types I and III by human fibroblasts derived from post-burn hypertrophic scars and from normal skin.  

Science.gov (United States)

Fibroblasts are thought to be partially responsible for the persisting contractile forces that result in burn contractures. Using a monolayer cell culture and fibroblast populated collagen lattice (FPCL) three-dimensional model we subjected hypertrophic scar and non-cicatricial fibroblasts to the antifibrogenic agent pentoxifylline (PTF - 1mg/mL) in order to reduce proliferation, collagen types I and III synthesis and model contraction. Fibroblasts were isolated from post-burn hypertrophic scars (HSHF) and non-scarred skin (NHF). Cells were grown in monolayers or incorporated into FPCL's and exposed to PTF. In monolayer, cell number proliferation was reduced (46.35% in HSHF group and 37.73% in NHF group, pFPCL. PMID:19303218

Isaac, Cesar; Mathor, Mônica Beatriz; Bariani, Giovani; Paggiaro, André Oliveira; Herson, Marisa Roma; Goldenstein-Schainberg, Claudia; Carrasco, Solange; Teodoro, Walcy Rosolia; Yoshinari, Natalino Hajime; Ferreira, Marcus Castro

2009-08-01

212

Repair of sublethal damage in diploid human fibroblasts: a comparison between human and rodent cell lines  

International Nuclear Information System (INIS)

The repair of sublethal damage has been compared in the human cell lines GM-498 and GM 3440 with that which occurs in the rodent lines CHO and V-79. The data suggests that the human cell lines repair sublethal damage at least as well as the rodent cell lines. Another object of this investigation was to determine if the repair of sublethal damage can be observed in the initial region of the survival curve. While repair of sublethal damage can be observed in the second decade, it cannot be observed in the first, assuming no redistribution in the cell cycle. 12 references, 6 figures

1983-07-01

213

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 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.

2010-06-15

214

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

215

Epidermal growth factor (EGF) antagonizes transforming growth factor (TGF)-beta1-induced collagen lattice contraction by human skin fibroblasts.  

Science.gov (United States)

Wound contraction plays an important role in healing, but in extreme conditions, it may lead to excessive scar formation and pathological wound contracture. To date, the key regulator of excessive contracture is known to be transforming growth factor-beta (TGF-beta1). In this study, we have evaluated epidermal growth factor (EGF) antagonism in fibroblast-populated collagen lattice (FPCL) gel contraction, which has been generally used as an in vitro model thought to mimic wound contraction in vivo. As expected, TGF-beta1 treatment enhanced normal fibroblast-induced collagen gel contraction in a dose-dependent manner. In contrast, EGF did not affect normal gel formation, but significantly antagonized TGF-beta1-induced gel formation (p<0.05 at 100 ng/ml), whereas the other growth factor, platelet-derived growth factor (PDGF), did not altered either normal or TGF-beta1-induced gel contractions. Similarly, EGF treatment, but not PDGF, also significantly suppressed TGF-beta1 release that was autologously elicited by TGF-beta1 treatment (p<0.01 at 100 ng/ml). Therefore, the results suggest that EGF may negatively regulate the role of TGF-beta1 through attenuating autologous release of TGF-beta1. PMID:11145189

Park, J S; Kim, J Y; Cho, J Y; Kang, J S; Yu, Y H

2000-12-01

216

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.

217

Selective expression of a normal action of the 1,25-dihydroxyvitamin D3 receptor in human skin fibroblasts with hereditary severe defects in multiple actions of that receptor.  

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We evaluated three actions of 1,25-dihydroxycholecalciferol [1,25-(OH)2D3] in human skin fibroblasts to test for heterogeneity in hormone-response coupling. In fibroblasts from normal subjects the 1,25-(OH)2D3 concentrations for half-maximal effect (EC50) were: for mitogenic effect 0.0001-0.0005 nM, for antimitogenic effect 1 nM, and for induction of 25-OHD3 24-hydroxylase (24-OHase) 5 nM. To evaluate the effects of mutations presumed to be in the gene for the 1,25-(OH)2D3 receptor we examine...

1989-01-01

218

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

1988-01-01

219

Ultraviolet A radiation induces immediate release of iron in human primary skin fibroblasts: The role of ferritin  

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In mammalian cells, the level of the iron-storage protein ferritin (Ft) is tightly controlled by the iron-regulatory protein-1 (IRP-1) at the posttranscriptional level. This regulation prevents iron acting as a catalyst in reactions between reactive oxygen species and biomolecules. The ultraviolet A (UVA) radiation component of sunlight (320–400 nm) has been shown to be a source of oxidative stress to skin via generation of reactive oxygen species. We report here that the exposure of human ...

Pourzand, Charareh; Watkin, Richard D.; Brown, Jonathan E.; Tyrrell, Rex M.

1999-01-01

220

Effect of fibroblast growth factor saporin mitotoxins on human bladder cell lines.  

Science.gov (United States)

Mitotoxins targeted via high-affinity growth factor receptors on the cell surface are a potential means of anticancer therapy. We have evaluated the effect of a chemically conjugated (FGF2-SAP) and a fusion protein (rFGF2-SAP) mitotoxin containing FGF-2 and saporin on normal (FHs 738B1) and malignant bladder cell lines (HT1197, TCCSUP, EJ-6, and RT4). The FGF-saporins demonstrated potent cytotoxicity in malignant bladder cell lines with an ID50 range of 0.13-13.6 nM, whereas cells derived from normal fetal bladder (FHs 738B1) were less sensitive to FGF2-saporins (ID50 > 100 nM). Greater than a 100-fold difference in cytotoxicity between FGF-saporins and unconjugated saporin was observed. Assessment of cellular FGF-2 content and secretion showed that FHs 738B1 and TCCSUP contained and secreted significantly more FGF-2 compared to other cell lines tested. (125)I-FGF-2 receptor binding studies showed the presence of high-affinity (pM) FGF receptors on all bladder cell lines. Cross-linking studies revealed the presence of a major receptor-ligand complex of 90 kDa on FHs 738B1 and 160-170 kDa on the other bladder cell lines. All cell lines studied, except RT4, expressed solely FGFR-1. These studies demonstrate that FGF2-saporins have antiproliferative activity on human bladder cancer cell lines. However, the number of high-affinity FGF receptors, and FGF-2 cellular content and secretion are not absolute determinants of cellular sensitivity to FGF2-saporins. PMID:9344046

Tetzke, T A; Caton, M C; Maher, P A; Parandoosh, Z

1997-11-01

 
 
 
 
221

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

222

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.

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

2014-01-01

223

Control by fibroblast growth factor of differentiation in the BC3H1 muscle cell line  

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The regulation of creatine phosphokinase (CPK) expression by polypeptide growth factors has been examined in the clonal mouse muscle BC3H1 cell line. After arrest of cell growth by exposure to low concentrations of serum, BC3H1 cells accumulate high levels of muscle- specific proteins including CPK. The induction of this enzyme is reversible in the presence of high concentrations of fetal calf serum, which cause quiescent, differentiated cells to reenter the cell cycle. Under these conditions...

1985-01-01

224

Contributions of differential p53 expression in the spontaneous immortalization of a chicken embryo fibroblast cell line  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background The present study was carried out to determine whether the p53 pathway played a role in the spontaneous immortalization of the SC-2 chicken embryo fibroblast (CEF cell line that has been in continuous culture for over three years. Results The SC-2 cell line emerged from an extended crisis period with a considerably slower growth rate than primary CEF cells. The phenotype of the SC-2 cells changed dramatically at about passage 80, appearing smaller than at earlier passages (e.g., passage 43 and possessing a small, compact morphology. This morphological change coincided with an increase in growth rate. Passage 43 SC-2 cells expressed undetectable levels of p53 mRNA, but by passage 95, the levels were elevated compared to primary passage 6 CEF cells and similar to levels in senescent CEF cells. However, the high level of p53 mRNA detected in passage 95 SC-2 cells did not correlate to functional protein activity. The expression levels of the p53-regulated p21WAF1 gene were significantly decreased in all SC-2 passages that were analyzed. Examination of the Rb pathway revealed that E2F-1 and p15INK4b expression fluctuated with increasing passages, with levels higher in passage 95 SC-2 cells compared to primary passage 6 CEF cells. Conclusion The present study suggests that altered expression of genes involved in the p53 and Rb pathways, specifically, p53 and p21WAF1, may have contributed to the immortalization of the SC-2 CEF cell line.

Kim Hyunggee

2006-06-01

225

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)

1989-01-01

226

Effect of sulphur mustard on human skin cell lines with differential agent sensitivity.  

Science.gov (United States)

The ability of sulphur mustard (HD) to induce DNA damage places limits on the efficacy of approaches aimed at protecting human cells from the cytotoxic effects of HD using a variety of protective agents such as thiol-containing esters and protease inhibitors. In the present study, potential alternative strategies were investigated by examining the differential effects of HD on G361, SVK14, HaCaT and NCTC 2544 human skin cells. The G361 cell line was more resistant to the cytotoxic effects of HD than the NCTC, HaCaT and SVK14 cell lines at HD doses of >3 and G361 and SVK14 cell culture viability. Buthionine sulphoximine (BSO) pretreatment increased the sensitivity of all four cell lines to HD. A substantial proportion of the resistance of G361 cells to HD was attributable to BSO-mediated effects on antioxidant-mediated metabolism, although G361 cultures still retained a high degree of viability at 30 microM HD following BSO pretreatment. Cell cycle analysis confirmed that SVK14 cells were relatively more sensitive to HD, as shown by the 2.1-fold reduction (P G361 cells in G0/G1 phase following HD exposure, suggesting the existence of a more efficient G0/G1 checkpoint control mechanism in this cell line. Manipulation of the cell cycle using various modulating agents did not increase the resistance of cell lines to the cytotoxic effects of HD. PMID:15747377

Simpson, Rachel; Lindsay, Christopher D

2005-01-01

227

Kiwi fruit (Actinidia chinensis L.) polysaccharides exert stimulating effects on cell proliferation via enhanced growth factor receptors, energy production, and collagen synthesis of human keratinocytes, fibroblasts, and skin equivalents.  

Science.gov (United States)

Within physiological engineering exogenous carbohydrates were recently confirmed as pharmacologically active compounds. To investigate potential dermatological activity purified polysaccharides from kiwi fruits (Actinidia chinensis L., Actinidiaceae) were characterized concerning monomer composition, linkage types and molecular weights and were tested under in vitro conditions for regulating activities on cell physiology of human keratinocytes, fibroblasts, and skin equivalents. Ten micrograms per milliliter of raw polysaccharide, neutral type-II-arabinogalactans, and acidic arabinorhamnogalacturonans of kiwi fruits stimulated cell proliferation of human keratinocytes (NHK, HaCaT) up to 30% significantly while mitochondrial activity was stimulated for nearly 25% in regard to control cells. Fibroblasts were not as sensitive as keratinocytes but >130 microg/ml kiwi fruit polysaccharides increased proliferation and ATP-synthesis significantly, too. Proliferation-stimulating activity was dependent on terminal 1-alpha-l-arabinose residues since enzymatic release of these sugar moieties caused significantly decreased proliferation of HaCaT and fibroblasts of about 10% in regard to untreated cells. In three dimensional skin equivalents, it was shown that the polysaccharides led to a doubled collagen synthesis of fibroblasts compared to the normally strongly reduced biosynthetic activity. PMID:15389574

Deters, Alexandra M; Schröder, Klaus R; Hensel, Andreas

2005-03-01

228

Parthenolide inhibits proliferation of fibroblast-like synoviocytes in vitro.  

Science.gov (United States)

Parthenolide is a bioactive constituent of an aromatic herb Feverfew (Tanacetum parthenium). It has been found that both parthenolide and extract of feverfew have anti-inflammatory and antinociceptive properties. Moreover, they demonstrate antiproliferative activities on different human tumour cells. The massive hyperplasia of synovial fibroblasts is the one of the most striking features of rheumatoid arthritis. It is not known whether this is due to the proliferation of synovial fibroblasts or to defective apoptosis. We investigated the effect of parthenolide on the proliferation of rabbit synoviocytes cell line HIG-82, rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) and human skin fibroblasts (HSF) in vitro. Cell proliferation was assessed by means of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and 5'-bromo-2'-deoxy-uridine methods. Parthenolide inhibited proliferation of HIG-82 and human RA-FLS. The proliferation of HSF was inhibited less effectively. The antiproliferative potential of parthenolide was demonstrated. PMID:18568393

Parada-Turska, Jolanta; Mitura, Agata; Brzana, Wojciech; Jab?o?ski, Miros?aw; Majdan, Maria; Rzeski, Wojciech

2008-08-01

229

Formation of DNA single-strand breaks by near-ultraviolet and gamma-rays in normal and Bloom's syndrome skin fibroblasts  

International Nuclear Information System (INIS)

The formation of single-strand breaks by near-ultraviolet light at 313 nm and by aerobic gamma-rays was compared for skin fibroblast monolayer cultures from 4 normal donors (NF) and 8 patients with Bloom's syndrome (BS) by the alkaline elution method. In 6 of 8 BS strains, the number of breaks induced by near-ultraviolet light, 2.25 kJ/sq m, at 0 degrees was comparable to NF, while elevated breakage was observed in BS strains HG 369 and HG 916. Breakage frequencies were increased substantially in 6 of 8 BS strains relative to NF when the near-ultraviolet light exposure was at 37 degrees. BS strain GM 2520 represents an exception since normal breakage frequencies were induced both at 0 degrees and 37 degrees. Aerobic gamma-rays (75 R) induced comparable numbers of single-strand breaks in BS and NF strains at 0 degrees. The breakage frequencies were reduced an average of 17% in NF when the same dose was given at 30 degrees followed by 6 min incubation. Under the same conditions, the breakage frequencies were on the average reduced by 42% relative to 0 degrees in the BS strains, indicating that they possess normal or possibly slightly increased capacities for the rejoining of gamma-ray-induced breaks

1981-01-01

230

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

231

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

1995-01-01

232

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

Energy Technology Data Exchange (ETDEWEB)

A total of 153 hprt mutants (23 spontaneous, 130 {gamma}-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 {gamma}-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 {gamma}-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{prime} end of the hprt gene. 46 refs., 4 figs., 2 tabs.

Park, M.S.; Hanks, T.; Jaberaboansari, A.; Chen, D.J. [Los Alamos National Lab., NM (United States)

1995-01-01

233

Differential modulation of CXCR4 and CD40 protein levels by skin sensitizers and irritants in the FSDC cell line  

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The development of non-animal methods for skin sensitization testing is an urgent challenge. Some of the most promising in vitro approaches are based on the analysis of phenotypical and functional modifications induced by sensitizers in dendritic cell models. In this work, we evaluated, for the first time, a fetal skin-derived dendritic cell line (FSDC) as a model to discriminate between sensitizers and irritants, through analysis of their effects on CD40 and CXCR4 protein expression. The che...

Neves, Bruno Miguel; Cruz, Maria Teresa; Francisco, Vera; Gonc?alo, Margarida; Figueiredo, Ame?rico; Duarte, Carlos B.; Lopes, Maria Celeste

2008-01-01

234

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

235

Protective effect of enzymatic hydrolysates from highbush blueberry (Vaccinium corymbosum L.) against hydrogen peroxide-induced oxidative damage in Chinese hamster lung fibroblast cell line  

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Blueberry was enzymatically hydrolyzed using selected commercial food grade carbohydrases (AMG, Celluclast, Termamyl, Ultraflo and Viscozyme) and proteases (Alcalase, Flavourzyme, Kojizyme, Neutrase and Protamex) to obtain water soluble compounds, and their protective effect was investigated against H2O2-induced damage in Chinese hamster lung fibroblast cell line (V79-4) via various published methods. Both AMG and Alcalase hydrolysates showed higher total phenolic content as well as higher ce...

2010-01-01

236

Estimation of the breast skin-line in mammograms using multidirectional Gabor filters.  

Science.gov (United States)

Segmentation of the breast region is a fundamental step in any system for computerized analysis of mammograms. In this work, we propose a novel procedure for the estimation of the breast skin-line based upon multidirectional Gabor filtering. The method includes an adaptive values-of-interest (VOI) transformation, extraction of the skin-air ribbon by Otsu's thresholding method and the Euclidean distance transform, Gabor filtering with 18 real kernels, and a step for suppression of false edge points using the magnitude and phase responses of the filters. On a test set of 361 images from different acquisition modalities (screen-film and full-field digital mammograms), the average Hausdorff and polyline distances obtained were 2.85 mm and 0.84 mm, respectively, with reference to the ground-truth boundaries provided by an expert radiologist. When compared with the results obtained by other state-of-the-art methods on the same set of images and with respect to the same ground-truth boundaries, our method mostly outperformed the other approaches. The results demonstrate the effectiveness and robustness of the proposed algorithm. PMID:24209932

Casti, P; Mencattini, A; Salmeri, M; Ancona, A; Mangeri, F; Pepe, M L; Rangayyan, R M

2013-11-01

237

The levels of vascular smooth as well as skeletal muscle actin mRNAs differ substantially among both myoblast and fibroblast lines with different skeletal myogenic potentials.  

Science.gov (United States)

Little is known about the factors which regulate vascular smooth muscle (vsm) actin gene expression during skeletal myogenesis in culture. We have therefore looked for differences in the levels of accumulation of vsm actin mRNA among six mouse cell lines differing in apparent myogenic potential or in the complement of myogenesis determination genes which they express: NIH 3T3 and 10T1/2 non-myogenic fibroblasts and four myogenic lines--3T3-MyoD1 and 10EMc11s, MyoD/myogenin expressing sublines of the fibroblast lines, derived by transfer into the parent lines of a MyoD cDNA expression construct; C2C12, which expresses all four known myogenesis determination genes; and BC3H1, which expresses myf-5, myogenin, little herculin, and no MyoD. In differentiated cells of all four myogenic lines, vsm actin mRNA was expressed at levels dramatically higher than in growth-arrested NIH 3T3 cells, consistent with expression of vsm actin mRNA as an intrinsic part of the skeletal myogenic program somehow directed by myogenesis determination gene products. Interestingly, however, the level of vsm actin mRNA in growth arrested C3H10T1/2 fibroblasts was also dramatically higher than that in NIH 3T3. In view of these findings, and of the relative ease with which 10T1/2 as opposed to NIH 3T3 cells can be converted to myogenic lines, we hypothesize that factors which can act to regulate vsm actin gene expression in the absence of myogenesis determination gene expression may also influence the skeletal myogenic potential of the cells in which they are found. Among the myogenic lines, the ratio of vsm to skm actin mRNA was highest in BC3H1 cells, raising the possibility that were these cells forced to express MyoD and/or more herculin, as do the other myogenic lines, the ratio would decrease. Thus both fibroblast and myogenic lines will be useful for investigating the mechanisms controlling skeletal myogenesis and vsm and skm actin gene expression during myogenesis. PMID:1477607

Sharp, S B; Vazquez, A; Theimer, M; Silva, D K; Muscati, S R; Sylber, M; Mogassa, M

1992-12-01

238

Murine schistosomiasis: selective inhibition in vitro of fibroblast collagen production by mononuclear cells.  

Science.gov (United States)

Primary cell cultures from the livers of mice infected with Schistosoma mansoni were prepared and cells with the appearance of fibroblasts by light microscopy were isolated. Collagen synthesis was estimated by measuring incorporation of 14C-proline into collagenase-sensitive proteins for both culture media and cell layers. Coculture of splenic T cells from infected mice with these hepatic fibroblasts caused greater selective and specific reduction in collagen production than did coculture using spleen cells from normal mice. There was a parallel inhibition in collagen within the cell layer which indicates that the marked decrease in collagen production was due to inhibition of synthesis and not related to changes in solubility or secretion. Primary culture of mouse skin fibroblasts showed similar responses to coculture but an established fibroblast line, 3T3, was unresponsive. Inflammatory cells appear to influence hepatic fibroblasts isolated under our experimental condition in several ways, such as opposite effects on collagen synthesis and cell proliferation. PMID:3124644

Mansour, M M; Dunn, M A; Salah, L A; Woody, J N

1988-01-01

239

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

240

Monoclonal antibody to the type I insulin-like growth factor (IGF-I) receptor blocks IGF-I receptor-mediated DNA synthesis: clarification of the mitogenic mechanisms of IGF-I and insulin in human skin fibroblasts  

International Nuclear Information System (INIS)

Insulin and insulin-like growth factor type I (IGF-I) stimulate an overlapping spectrum of biological responses in human skin fibroblasts. Although insulin and IGF-I are known to stimulate the incorporation of [3H]thymidine into DNA in these cells, the identify of the receptor(s) that mediates this effect has not been fully clarified. The mouse anti-human IGF-I receptor antibody ?IR-3 binds with specificity to IGF-I but not to insulin receptors in human placental membranes; it also specifically inhibits the binding of 125I-labeled IGF-I but not 125I-labeled insulin to suspensions of human skin fibroblasts in a dose-dependent manner. ?IR-3 competitively inhibits IGF-I-mediated stimulation of [3H]thymidine incorporation into DNA. This inhibition is dependent on the concentration of ?IR-3 and in the presence of a fixed antibody concentration can be partially overcome by high concentrations of IGF-I. In contrast, at concentrations of 3H]thymidine incorporation is not inhibited by ?IR-3. However, the incremental effects of higher concentrations (> 1 ?g/ml) of insulin on [3H]thymidine incorporation are inhibited by ?IR-3. ?IR-3 is a highly specific antagonist of IGF-I receptor-mediated mitogenesis in human skin fibroblasts. By using this antibody, it is shown directly that insulin can act through the IGF-I receptor to stimulate DNA synthesis but can also activate this effect through the insulin receptor itself

1986-01-01

 
 
 
 
241

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

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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

242

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)

1998-01-01

243

Human T-Cell Leukemia Virus Type 2 (HTLV-2) Tax Protein Transforms a Rat Fibroblast Cell Line but Less Efficiently than HTLV-1 Tax  

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Human T-cell leukemia virus type 1 (HTLV-1) and HTLV-2 are retroviruses with similar biological properties. Whereas HTLV-1 is the causative agent of an aggressive T-cell leukemia, HTLV-2 has been associated with only a few cases of lymphoproliferative disorders. Tax1 and Tax2 are the transcriptional activators of HTLV-1 and HTLV-2, respectively. Here we show that Tax2 transformed a Rat-1 fibroblast cell line to form colonies in soft agar, but the size and number of the colonies were lower tha...

Endo, Keiichi; Hirata, Akira; Iwai, Kousuke; Sakurai, Mamoru; Fukushi, Masaya; Oie, Masayasu; Higuchi, Masaya; Hall, William W.; Gejyo, Fumitake; Fujii, Masahiro

2002-01-01

244

The hallmarks of fibroblast ageing.  

Science.gov (United States)

Ageing is influenced by the intrinsic disposition delineating what is maximally possible and extrinsic factors determining how that frame is individually exploited. Intrinsic and extrinsic ageing processes act on the dermis, a post-mitotic skin compartment mainly consisting of extracellular matrix and fibroblasts. Dermal fibroblasts are long-lived cells constantly undergoing damage accumulation and (mal-)adaptation, thus constituting a powerful indicator system for human ageing. Here, we use the systematic of ubiquitous hallmarks of ageing (Lopez-Otin et al., 2013, Cell 153) to categorise the available knowledge regarding dermal fibroblast ageing. We discriminate processes inducible in culture from phenomena apparent in skin biopsies or primary cells from old donors, coming to the following conclusions: (i) Fibroblasts aged in culture exhibit most of the established, ubiquitous hallmarks of ageing. (ii) Not all of these hallmarks have been detected or investigated in fibroblasts aged in situ (in the skin). (iii) Dermal fibroblasts aged in vitro and in vivo exhibit additional features currently not considered ubiquitous hallmarks of ageing. (iv) The ageing process of dermal fibroblasts in their physiological tissue environment has only been partially elucidated, although these cells have been a preferred model of cell ageing in vitro for decades. PMID:24686308

Tigges, Julia; Krutmann, Jean; Fritsche, Ellen; Haendeler, Judith; Schaal, Heiner; Fischer, Jens W; Kalfalah, Faiza; Reinke, Hans; Reifenberger, Guido; Stühler, Kai; Ventura, Natascia; Gundermann, Sabrina; Boukamp, Petra; Boege, Fritz

2014-06-01

245

Cytotoxicity and Proliferation Studies with Arsenic in Established Human Cell Lines: Keratinocytes, Melanocytes, Dendritic Cells, Dermal Fibroblasts, Microvascular Endothelial Cells, Monocytes and T-Cells  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract: Based on the hypothesis that arsenic exposure results in toxicity and mitogenecity, this study examined the dose-response of arsenic in established human cell lines of keratinocytes (HaCaT, melanocytes (1675, dendritic cells (THP-1/A23187, dermal fibroblasts (CRL1904, microvascular endothelial cells (HMEC, monocytes (THP-1, and T cells (Jurkat. Cytotoxicity was determined by incubating THP-1, THP-1+ A23187 and JKT cells in RPMI 1640, 1675 in Vitacell, HMEC in EBM, and dermal fibroblasts and HaCaT in DMEM with 10% fetal bovine serum, 1% streptomycin and penicillin for 72 hrs in 96-well microtiter plates, at 37oC in a 5% CO2 incubator with different concentrations of arsenic using fluorescein diacetate (FDA. Cell proliferation in 96-well plates was determined in cultured cells starved by prior incubation for 24 hrs in 1% FBS and exposed for 72 hours, using the 96 cell titer proliferation solution (Promega assay. Cytotoxicity assays yielded LD50s of 9 μg/mL for HaCaT, 1.5 μg/mL for CRL 1675, 1.5 μg/mL for dendritic cells, 37 μg/mL for dermal fibroblasts, 0.48 μg/mL for HMEC, 50 μg/mL for THP-1 cells and 50 μg/mL for JKT-T cells. The peak proliferation was observed at 6 μg/mL for HaCaT and THP-1 cells, 0.19 μg/mL for CRL 1675, dendritic cells, and HMEC, and 1.5 μg/mL for dermal fibroblasts and Jurkat T cells. These results show that arsenic is toxic at high doses to keratinocytes, fibroblasts, monocytes and T cells, and toxic at lower doses to melanocytes, microvascular endothelial cells and dendritic cells. Proliferation studies showed sub-lethal doses of arsenic to be mitogenic.

Hari H. P. Cohly

2003-01-01

246

Development of an artificial lock for the skin-pass section in a hot dip galvanising line  

International Nuclear Information System (INIS)

In this paper, we present the application of data mining techniques to develop an artificial lock for the skin-pass in an attempt to solve a problem that can arise during the galvanising manufacturing process:the wrong labelling of the steel grade of a coil. In order to detect these errors and thus to avoid that coils with different properties than expected end up with a client, we propose neural network-based models for on-line predicting the strip elongation in the skin-pass section according to the manufacturing conditions and its chemical composition. thus, a significant difference between estimated and measured elongation would mean that the coil must be removed from the line for further analyses. (Author) 14 refs

2008-01-01

247

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

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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 ...

2011-01-01

248

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

2004-12-01

249

Smooth muscle differentiation in scleroderma fibroblastic cells.  

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Using antibodies to alpha-smooth muscle actin and desmin on paraffin-embedded formalin-fixed tissue sections, the authors demonstrate that fibroblastic cells of localized and systemic scleroderma lesions express features of smooth muscle differentiation. Eleven of eleven skin specimens of systemic sclerosis patients and two of four skin specimens of localized scleroderma displayed the presence of fibroblasts expressing alpha-smooth muscle actin, a cell population that predominated in areas of...

Sappino, A. P.; Masouye?, I.; Saurat, J. H.; Gabbiani, G.

1990-01-01

250

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.

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

251

Contact sensitizers downregulate the expression of the chemokine receptors CCR6 and CXCR4 in a skin dendritic cell line  

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Chemokines are involved in the control of dendritic cell (DC) trafficking, which is critical for the immune response, namely in allergic contact dermatitis (ACD). In this work, we investigated by flow cytometry the effect of the contact sensitizers 2,4-dinitrofluorobenzene (DNFB), 1,4-phenylenediamine (PPD) and nickel sulfate (NiSO(4)), on the surface expression of the chemokine receptors CCR6 and CXCR4 in DC. As an experimental model of a DC we used a fetal skin-derived dendritic cell line (...

Cruz, Mt; Gonc?alo, Margarida; Paiva, A.; Morgado, Jm; Figueiredo, A.; Duarte, Cb; Lopes, Mc

2005-01-01

252

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

Directory of Open Access Journals (Sweden)

Full Text Available 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 reconstitution and thus the precise analysis of growth regulatory mechanisms.

Stark Hans-Jürgen

2004-01-01

253

Application of collagen-chitosan/fibrin glue asymmetric scaffolds in skin tissue engineering*  

Digital Repository Infrastructure Vision for European Research (DRIVER)

To create a scaffold that is suitable for the construction of tissue-engineered skin, a novel asymmetric porous scaffold with different pore sizes on either side was prepared by combining a collagen-chitosan porous membrane with fibrin glue. Tissue-engineered skin was fabricated using this asymmetric scaffold, fibroblasts, and a human keratinocyte line (HaCaT). Epidermal cells could be seen growing easily and achieved confluence on the fibrin glue on the upper surface of the scaffold. Scannin...

Han, Chun-mao; Zhang, Li-ping; Sun, Jin-zhang; Shi, Hai-fei; Zhou, Jie; Gao, Chang-you

2010-01-01

254

Characterisation of human tumour cell lines using antibodies to intermediate filaments.  

Science.gov (United States)

The human hepatoma cell line G2 (Hep G2) has been compared to lung carcinoma, sarcoma and skin fibroblasts for the expression of intermediate filaments, i.e. vimentin and cytokeratin. The immunofluorescence study revealed that, in contradiction to Szecheng et al. (1987), cytokeratin and vimentin are absent in Hep G2. Human skin fibroblasts and sarcoma cells expressed vimentin as expected for their mesenchymal origin, but a positive reaction to vimentin could also be shown in lung carcinoma cells. However, the vimentin filament structure of both these tumour cell lines was different in comparison with skin fibroblasts. Therefore determining the exact tissue origin of tumour cell lines by means of intermediate filament characterization remains doubtful. PMID:7692876

Moorthi, C; Willers, I; Ressler, B; Goedde, H W

1993-01-01

255

PDGF-induced receptor phosphorylation and phosphoinositide hydrolysis are unaffected by protein kinase C activation in mouse Swiss 3T3 and human skin fibroblasts.  

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Short (1-10 min) pretreatment of intact cells with activators of protein kinase C (e.g. phorbol-12 myristate, 13-acetate, PMA) affects the activity of a variety of surface receptors (for growth factors, hormones and neurotransmitters), with inhibition of transmembrane signal generation. In two types of fibroblasts we demonstrate that the PDGF receptor is unaffected by PMA. Exposure to PMA at concentrations up to 100 nM for 10 min failed to inhibit either one of the agonist-induced, receptor-c...

Comoglio, Paolo

1986-01-01

256

Differential susceptibilities of human lung, breast and skin cancer cell lines to killing by five sea anemone venoms  

Directory of Open Access Journals (Sweden)

Full Text Available Although sea anemones are well known for being rich sources of toxins, including cytolysins and neurotoxins, their venoms and toxins have been poorly studied. In the present study, the venoms from five sea anemones (Heteractis crispa, Heteractis magnifica, Heteractis malu, Cryptodendrum adhaesivum and Entacmaea quadricolor were obtained by the milking technique, and the potential of these venoms to kill cancer cells was tested on three cell lines (A549 lung cancer, T47D breast cancer and A431 skin cancer. The total protein level in the crude extract was determined by the bicinchoninic acid (BCA protein assay. The cytotoxicity on different cell lines was assayed using the 3-(4, 5-dimethylthiazol-2yl-2, 5-diphenyltetrazolium bromide (MTT assay which measures survival based on the detection of mitochondrial activity and by the crystal violet assay, which measures survival based on the ability of cells to remain adherent to microplates. The results indicate that the sea anemone venom is cytotoxic to human cancer cells. The A549 cell line was the most sensitive of the cell lines tested with a significant reduction in viability observed at 40 µg/mL. H. malu, C. adhaesivum and E. quadricolor had a significant inhibitory effect on A431 cells. Furthermore, H. malu and C. adhaesivum had a significant inhibitory effect on T47D cell line at 40 µg/mL. In conclusion, the sea anemone venoms tested have the potential to be developed as anticancer agents.

M Ramezanpour

2012-01-01

257

Evaluation of the effect of laser radiation on fibroblast proliferation in repair of skin wounds of rats with iron deficiency anemia  

Science.gov (United States)

The aim of this study was to assess the effect of low- level laser therapy (LLLT) on fibroblast proliferation on wound repair of rats with Iron deficiency anemia since there is no reports on literature about this subject. Iron deficiency anemia was induced on 36 newborn rats then an excisional wound was created on the dorsum of the animals which were divided into four groups: (I) - non-anemic, (II) - Anemic, (III) - non-anemic + LLLT, (IV) Anemic+ LLLT. The animals in each group were sacrificed at 7, 14 and 21 days. Laser irradiation was performed on each group (?660nm,40Mw,CW) by contact mode with a dose of 2,5J/ cm2 in four points on the area of the wound and total of 10J/cm2 per session. Data were evaluated by analysis of variance (ANOVA) followed by Paired t-test. The results showed LLLT was able to stimulate fibroblastic proliferation in rats with iron deficiency anemia at the 21st day while at control group (III) no statistically significant differences was found.

Decastro, Isabele C. V.; Oliveira-Sampaio, Susana C. P.; Monteiro, Juliana S. De C.; Ferreira, Maria De Fátima L.; Cangussu, Maria T.; N. Dos Santos, Jean; Pinheiro, Antonio Luiz B.

2011-02-01

258

Characterization of a novel fibroblast growth factor 10 (Fgf10) knock-in mouse line to target mesenchymal progenitors during embryonic development.  

Science.gov (United States)

Fibroblast growth factor 10 (Fgf10) is a key regulator of diverse organogenetic programs during mouse development, particularly branching morphogenesis. Fgf10-null mice suffer from lung and limb agenesis as well as cecal and colonic atresia and are thus not viable. To date, the Mlcv1v-nLacZ-24 transgenic mouse strain (referred to as Fgf10(LacZ)), which carries a LacZ insertion 114 kb upstream of exon 1 of Fgf10 gene, has been the only strain to allow transient lineage tracing of Fgf10-positive cells. Here, we describe a novel Fgf10(Cre-ERT2) knock-in line (Fgf10(iCre)) in which a Cre-ERT2-IRES-YFP cassette has been introduced in frame with the ATG of exon 1 of Fgf10 gene. Our studies show that Cre-ERT2 insertion disrupts Fgf10 function. However, administration of tamoxifen to Fgf10(iCre); Tomato(flox) double transgenic embryos or adult mice results in specific labeling of Fgf10-positive cells, which can be lineage-traced temporally and spatially. Moreover, we show that the Fgf10(iCre) line can be used for conditional gene inactivation in an inducible fashion during early developmental stages. We also provide evidence that transcription factors located in the first intron of Fgf10 gene are critical for maintaining Fgf10 expression over time. Thus, the Fgf10(iCre) line should serve as a powerful tool to explore the functions of Fgf10 in a controlled and stage-specific manner. PMID:22719891

El Agha, Elie; Al Alam, Denise; Carraro, Gianni; MacKenzie, Breanne; Goth, Kerstin; De Langhe, Stijn P; Voswinckel, Robert; Hajihosseini, Mohammad K; Rehan, Virender K; Bellusci, Saverio

2012-01-01

259

The role of first line of defence mechanisms in the pathogenesis of cellulitis in broiler chickens: skin structural, physiological and cellular response factors.  

Science.gov (United States)

The present study examined several basic attributes of first-line defence mechanisms in the skin as potential factors that may explain the susceptibility of broiler chickens to cellulitis. The variables including structural characteristics of the skin, physicochemical properties and cellular responses to the challenge with pathogens were compared between two categories of chickens, a strain of fast-growing commercial broiler chickens (susceptible to cellulitis) and leghorn chickens (resistant to cellulitis). There were substantial differences between leghorns and broilers with regard to physiological characteristics of the skin. Broiler skin was more amenable to injury and the wound-healing process was slow. Compared with leghorns, the lesions resulting from sub-dermal challenge in broilers were more severe and disseminated over a larger area. Mobilization of phagocytic cells (heterophils and macrophages) in leghorns was brisk even in the areas distant from the site of infection, whereas only few heterophils were recruited in the skin of broilers. The functional competence of heterophils in broilers was inferior when compared with leghorns. Based on the present finding, the predisposition of broilers to cellulitis appears to be primarily associated with the inferior first line of defence of their skin. Broilers in commercial situations may be at higher risk to succumb to even minor infection and eventually develop cellulitis because: (1) structural weaknesses of the skin may predispose broilers to skin injury and thus the risk of skin infection by pathogens is increased; (2) broiler skin surface is more likely to provide a conducive environment for colonization of Escherichia coli; (3) in the event of infection, poor recruitment of phagocytic cells to the site of infection may readily lead to widespread colonization of the tissue by pathogens causing cellulitis and (4) poor functional quality of the phagocytic cells that are mobilized compromise the ability of the host to contain the spread of infection. PMID:16300661

Olkowski, A A; Wojnarowicz, C; Chirino-Trejo, M; Wurtz, B M; Kumor, L

2005-12-01

260

Induction of matrix metalloproteinase-1 (MMP-1) during epidermal invasion of the stroma in human skin organ culture: keratinocyte stimulation of fibroblast MMP-1 production  

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Organ cultures of human skin were incubated for 8 days under growth factor-free conditions or exposed to 10?ng ml?1 of human recombinant epidermal growth factor (EGF) during the incubation period. Normal histological features were preserved in the absence of growth factor, while epithelial cells underwent a proliferative response and invaded the underlying stroma in the presence of exogenous EGF. The same concentrations of EGF that induced stromal invasion also resulted in up-regulation o...

Moon, S. E.; Dame, M. K.; Remick, D. R.; Elder, J. T.; Varani, J.

2001-01-01

 
 
 
 
261

Differentiation of a human leukemia cell line and expression of collagenase inhibitor.  

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A human collagenase inhibitor (CI) of Mr 28,500 has been extensively characterized in skin fibroblasts and identified in a variety of connective tissues. Because human alveolar macrophages synthesize and secrete both a collagenase and CI that are immunologically and functionally identical to their counterparts in fibroblasts, we studied the production of such proteins by an immature human cell line (HL60) that can be induced to differentiate along monocytic or granulocytic pathways. The cells...

1985-01-01

262

Differential susceptibilities of human lung, breast and skin cancer cell lines to killing by five sea anemone venoms  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Although sea anemones are well known for being rich sources of toxins, including cytolysins and neurotoxins, their venoms and toxins have been poorly studied. In the present study, the venoms from five sea anemones (Heteractis crispa, Heteractis magnifica, Heteractis malu, Cryptodendrum adhaesivum a [...] nd Entacmaea quadricolor) were obtained by the milking technique, and the potential of these venoms to kill cancer cells was tested on three cell lines (A549 lung cancer, T47D breast cancer and A431 skin cancer). The total protein level in the crude extract was determined by the bicinchoninic acid (BCA) protein assay. The cytotoxicity on different cell lines was assayed using the 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide (MTT) assay which measures survival based on the detection of mitochondrial activity and by the crystal violet assay, which measures survival based on the ability of cells to remain adherent to microplates. The results indicate that the sea anemone venom is cytotoxic to human cancer cells. The A549 cell line was the most sensitive of the cell lines tested with a significant reduction in viability observed at 40 µg/mL. H. malu, C. adhaesivum and E. quadricolor had a significant inhibitory effect on A431 cells. Furthermore, H. malu and C. adhaesivum had a significant inhibitory effect on T47D cell line at 40 µg/mL. In conclusion, the sea anemone venoms tested have the potential to be developed as anticancer agents.

M, Ramezanpour; K, Burke da Silva; BJ, Sanderson.

263

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.

G Rossi

2009-12-01

264

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

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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

265

Electronic waste leachate-mediated DNA fragmentation and cell death by apoptosis in mouse fibroblast (NIH/3T3) cell line.  

Science.gov (United States)

This study investigated the apoptotic effect of electronic waste on fibroblast cell line. Cells were treated with different concentrations of the leachate for 24h. Cell viability was detected by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test, nuclear morphology of cells was explored by acridine orange (AO)/ethidium bromide (EB) double staining, mitochondrial membrane potential was evaluated using JC-1 probe while cell cycle analysis was conducted using flow cytometry. The oxidative status was detected using DCFH-DA (dichlorofluorescin diacetate) probe and the relationship between cell death and ROS (reactive oxygen species) was investigated using N-acetylcysteine. Results showed an increased cell death as detected by MTT assay and AO/EB staining. Cell cycle analysis indicated an induction of sub/G1 events while JC-1 probe showed significant disruption of mitochondrial membrane potential. There was significant induction of ROS, while N-acetylcysteine protected the cells from DNA damage. These suggest apoptotic pathway as a possible mechanism of e-waste induced cyto-genotoxicity. PMID:23726292

Alabi, Okunola A; Bakare, Adekunle A; Filippin-Monteiro, Fabíola B; Sierra, Jelver A; Creczynski-Pasa, Tânia B

2013-08-01

266

Microprobe analysis of human fibroblasts  

International Nuclear Information System (INIS)

The Melbourne Proton Microprobe has been used to study the copper content in human skin fibroblast cells derived from patients with the genetic disease Menkes Syndrome. Both normal and diseased cells have been studied to investigate any elemental differences occurring between the two cell types. This paper details the preparatory techniques necessary for individual cell analysis and presents the elemental information with a new three dimensional contour mapping technique. These maps are used to highlight elemental differences between normal and mutant fibroblasts. The work also confirms the expected copper excess found in the Menkes cell and indicates that the microprobe can be used for rapid identification of a Menkes carrier

1985-11-01

267

Studies of molecular species of the human androgen receptor (AR): comparison of the physicochemical properties of the ["3H]methyltrienolone-AR complex formed in cytosol to the complex produced in intact genital skin fibroblasts  

International Nuclear Information System (INIS)

Two forms of the human genital skin fibroblast (GSF) androgen receptor (AR) complexed with ["3H]17 alpha-methyltrienolone were compared: 1) the intact complex formed in cytosol at 4 C (broken cell or B/C complex); and 2) the complex formed in the whole cell at 37 C (W/C complex). The intact form of the B/C complex was distinguished from partly degraded forms by the gel filtration profile in 0.5 M KCl. The W/C complex was considered to represent the transformed state of the receptor. The W/C complex had a smaller molecular radius than the B/C complex by gel filtration (Kav = 0.26-0.28 vs. 0.11-0.18). By low salt density gradient centrifugation, the B/C complex sedimented at 8.8S and the W/C complex at 6.6S. However, in 0.5 M KCl, each sedimented at 5.1S, and they were homogeneous, indicating that the monomeric forms differed markedly in molecular radius, but by only about 20,000 daltons in calculated mol wt (134,500 vs. 114,300 daltons). The complexes were separated from DNA, desalted, and compared by chromatography on DEAE-Sephacel and hydroxylapatite (HAP). The B/C complex bound readily to both column matrices and eluted from each as a sharp homogeneous peak: from DEAE at 172-190 mM KCl and from HAP at 123 mM phosphate. The W/C complex, however, was heterogeneous. One component did not bind to DEAE, and one eluted at 22-40 mM KCl. The W/C complex eluted from HAP as a peak at 42 mM, with a shoulder at 102 mM phosphate. Thus, transformation of the human genital skin fibroblast androgen receptor involves a major decrease in molecular radius and loss of negative charge with a possible loss of a 20,000-dalton macromolecular component

1986-01-01

268

BHK-21-derived cell lines that produce basic fibroblast growth factor, but not parental BHK-21 cells, initiate neuronal differentiation of neural crest progenitors.  

Science.gov (United States)

We present evidence that basic fibroblast growth factor (bFGF)-producing cells stimulate primary differentiation of neurons from neural crest progenitors. Baby hamster kidney (BHK-21) cells were stably cotransfected with plasmid pSV2/neo, which contains the gene conferring resistance to the neomycin analog G418 and expression vectors containing the human bFGF cDNA. Various clones, which differed in their bFGF production levels, were isolated. Homogeneous neural crest cells were cultured on monolayers of bFGF-producing, BHK-21-derived cell lines. While the parental BHK-21 cells, which do not produce detectable bFGF, had poor neurogenic ability, the various bFGF-producing clones promoted a 1.5- to 4-fold increase in neuronal cell number compared to the parental cells. This increase was correlated with the levels of bFGF produced by the different transfected clones, which ranged between 2.3 and 140 ng/mg protein. In contrast, no stimulation of neuronal differentiation was observed when neural crest cells were grown on monolayers of parental BHK cells transfected with plasmid pSV2/neo alone, or on a parental BHK-derived clone, which secretes high amounts of recombinant vascular endothelial growth factor (VEGF). Furthermore, the neuron-promoting ability of bFGF-producing cells could be mimicked by addition of exogenous bFGF to neural crest cells grown on the parental BHK line. A similar treatment of neural crest cells grown on laminin substrata, instead of BHK cells, resulted in increased survival of non-neuronal cells, but not of neurons (see also Kalcheim, C. 1989, Dev. Biol. 134, 1-10). Taken together, these results suggest that bFGF stimulates neuronal differentiation of neural crest cells by a cell-mediated signalling mechanism. PMID:1451657

Brill, G; Vaisman, N; Neufeld, G; Kalcheim, C

1992-08-01

269

Zinc Sulfate Failure as Accelerator of Collagen Biosynthesis and Fibroblast Proliferation.  

Science.gov (United States)

The effect of zinc sulfate on collagen biosynthesis and fibroblast proliferation has been studied in a tissue culture model system using human skin fibroblasts. Addition of zinc sulfate to newly established (low density) cultures in concentrations of 0.00...

J. C. Houck J. J. Amato M. D. Waters R. D. Moore

1971-01-01

270

Roles of the Ras-MEK-Mitogen-Activated Protein Kinase and Phosphatidylinositol 3-Kinase-Akt-mTOR Pathways in Jaagsiekte Sheep Retrovirus-Induced Transformation of Rodent Fibroblast and Epithelial Cell Lines  

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Jaagsiekte sheep retrovirus (JSRV) is the causative agent of ovine pulmonary adenocarcinoma (OPA), a transmissible lung cancer of sheep. The virus can induce tumors rapidly, and we previously found that the JSRV envelope protein (Env) functions as an oncogene, because it can transform mammalian and avian fibroblast cell lines. (N. Maeda, Proc. Natl. Acad. Sci. USA 98:4449-4454, 2001). The molecular mechanisms of JSRV Env transformation are of considerable interest. Several reports suggested t...

Maeda, Naoyoshi; Fu, Wuxia; Ortin, Aurora; Las Heras, Marcelo; Fan, Hung

2005-01-01

271

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  

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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 deficienc...

?ížková Alena; Stránecký Viktor; Ivánek Robert; Hartmannová Hana; Nosková Lenka; Piherová Lenka; Tesa?ová Markéta; Hansíková Hana; Honzík Tomáš; Zeman Ji?í; Divina Petr; Potocká Andrea; Paul Jan; Sperl Wolfgang; Mayr Johannes A

2008-01-01

272

An exploratory clinical study on the safety and efficacy of an autologous fibroblast-seeded artificial skin cultured with animal product-free medium in patients with diabetic foot ulcers.  

Science.gov (United States)

Cultured dermal substitutes have been used for the treatment of chronic skin ulcers; however, the biological risks of animal-derived materials in the culture process such as foetal bovine serum (FBS) have been reported. In this study, we prepared an autologous fibroblast-seeded artificial dermis (AFD) using animal-product-free medium supplemented with 2% patient autologous serum and without any animal-derived materials such as trypsin in the culturing process. We applied the AFD in five patients with diabetic ulcers and investigated its safety and efficacy. As the primary endpoint, we defined 'wound bed improvement' according to the percentage of granulation area to the whole wound area on day 21, and 60% or higher was regarded as improved. The mean age of the patients was 60·6 years and the mean duration of the ulcer was 22·6 months. In the evaluation of the primary endpoint, the 'wound bed' was improved in all patients [proportion of improvement: 100%, 95% confidence interval (CI): 48% to 100%]. Three patients had complete wound healing within 12 weeks after application and two patients had >80% wound healing at 12 weeks. Side effects were not serious. Our AFD may be a safe and effective treatment of diabetic ulcers. PMID:22958543

Morimoto, Naoki; Ito, Tatsuya; Takemoto, Satoru; Katakami, Mikiko; Kanda, Norikazu; Tada, Harue; Tanaka, Shiro; Teramukai, Satoshi; Kawai, Katsuya; Nakamura, Yoko; Kasai, Yasunari; Masayuki, Yokode; Maekawa, Taira; Shimizu, Akira; Suzuki, Shigehiko

2014-04-01

273

A correlation between ultraviolet-induced sister chromatid exchanges and ultraviolet-indced mutagenesis in ''Muntiacus muntjak'' (Indian Muntjac) skin fibroblasts in culture  

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The purpose of this thesis was to develop the capability of simultaneously assaying SCEs and mutations in Indian muntjac cells to determine (1) the relationship between the ultraviolet radiation (UVR) induction of SCEs and the UVR induction of mutations at the (UVR) HGPRT locus in Indian muntjac cells and (2) the possible role of DNA repair in the UVR induction of these two events. Indian muntjac skin fibroblasts were chosen for this study because of a unique karyotype consisting of a diploid chromosome number of 6 in females and 7 in males. An HGPRT mutation assay in Indian muntjac cells was developed by this author since at the time this study was undertaken no mutational assay system utilizing Indian muntjac cells existed. It is concluded from this study that a linear correlation exists betwen the UVR-induction of SCEs and of mutations to 6TG resistance in Indian muntjac cells. As more time is allowed between the UVR-induced DNA damage and onset of DNA replication, more of the lesions leading to both mutations and SCE formation are repaired. The fact that SCE and mutation frequencies are reduced at different rates may indicate that the lesions responsible for SCEs and for mutations are repaired differently.

Lugo, M.H.

1983-01-01

274

Fibroblast cultures in duchenne muscular dystrophy  

International Nuclear Information System (INIS)

Primary skin fibroblast cultures were grown from forearm pinch skin biopsies obtained from 24 patients with Duchenne muscular dystrophy (DMD) and ten normal controls matched for sex and age. The first subcultures were grown for 7 days and incubated with L-(3H)-proline for 24 hours. Intracellular collagen incoption was significantly decreased (2.2 X) and extracellular collagen incorporation significantly increased (1.8 X) in fibroblast cultures from patients with DMD by both collagenase assay and polyacrylamide gel electrophoresis. The synthesis of noncollagen proteins showed low values from the DMD fibroblast cultures. The alterations in synthesis and secretion of collagen and noncollagen proteins were characteristic only for the log phase of DMD fibroblasts. (author)

1977-01-01

275

Chondroitin sulfate and heparan sulfate-containing proteoglycans are both partners and targets of basic fibroblast growth factor-mediated proliferation in human metastatic melanoma cell lines.  

Science.gov (United States)

Basic fibroblast growth factor (FGF-2) and its respective tyrosine kinase receptors, form an autocrine loop that affects human melanoma growth and metastasis. The aim of the present study was to examine the possible participation of various glycosaminoglycans, i.e. chondroitin sulfate, dermatan sulfate and heparin on basal and FGF-2-induced growth of WM9 and M5 human metastatic melanoma cells. Exogenous glycosaminoglycans mildly inhibited WM9 cell's proliferation, which was abolished by FGF-2. Treatment with the specific inhibitor of the glycosaminoglycan sulfation, sodium chlorate, demonstrated that endogenous glycosaminoglycan/proteoglycan production is required for both basal and stimulated by FGF-2 proliferation of these cells. Heparin capably restored their growth, and unexpectedly exogenous chondroitin sulfate to WM9 and both chondroitin sulfate and dermatan sulfate to M5 cells allowed FGF-2 mitogenic stimulation. Furthermore, in WM9 cells the degradation of membrane-bound chondroitin/dermatan sulfate stimulates basal growth and even enhances FGF-2 stimulation. The specific tyrosine kinase inhibitor, genistein completely blocked the effects of FGF-2 and glycosaminoglycans on melanoma proliferation whereas the use of the neutralizing antibody for FGF-2 showed that the mitogenic effect of chondroitin sulfate involves the interaction of FGF-2 with its receptors. Both the amounts of chondroitin/dermatan/heparan sulfate and their sulfation levels differed between the cell lines and were distinctly modulated by FGF-2. In this study, we show that chondroitin/dermatan sulfate-containing proteoglycans, likely in cooperation with heparan sulfate, participate in metastatic melanoma cell FGF-2-induced mitogenic response, which represents a novel finding and establishes the central role of sulfated glycosaminoglycans on melanoma growth. PMID:17706452

Nikitovic, D; Assouti, M; Sifaki, M; Katonis, P; Krasagakis, K; Karamanos, N K; Tzanakakis, G N

2008-01-01

276

Active compound of Zingiber cassumunar Roxb. down-regulates the expression of genes involved in joint erosion in a human synovial fibroblast cell line.  

Science.gov (United States)

Rheumatoid arthritis (RA) is a chronic inflammatory disease of the synovium. It is involved in up-regulation of pro-inflammatory cytokines and matrix metalloproteinases (MMPs), resulting in joint inflammation and erosion. Zingiber cassumunar Roxb. has long been used to reduce joint pain and inflammation. This study aimed to investigate the inhibitory activities of an active compound of Z. cassumunar, (E)-4-(3',4'-dimethoxyphenyl)but-3-en-1-ol (compound D), against cytokine-induced up-regulation of catabolic genes involved in cartilage degradation in RA. Synovial fibroblast cell line, SW982, was cultured in media containing interleukin-1? (IL-1?), in the presence or absence of compound D at the concentration range of 1 to 100 µM. After 24 hours, the cells were analyzed for the expressions of MMPs, IL-1? and interleukin-1?-converting enzyme (ICE) by RT-PCR. MMPs activities in the culture media were analyzed by zymographic techniques. Dexamethasone was used as the positive control. It was found that compound D at the concentration of 10 - 100 µM significantly decreased the mRNA expressions of MMP-1, -2, -3, and -13 which was induced by IL-1? (P<0.05) concomitantly with a decrease in activities of these MMPs in the culture media. An increase in the mRNA expression of IL-1? and ICE was also suppressed by compound D. The results suggest that the potent activities of this compound may be involved in the reduction of IL-1? protein synthesis in both pro-form and active form which played an important role in up-regulation of MMPs. This study first revealed the chondroprotective activity of Z. cassumunar in the transcriptional level by suppressing cytokine-induced catabolic genes which caused cartilage erosion in RA. PMID:24082324

Chaiwongsa, Rujirek; Ongchai, Siriwan; Boonsing, Phorani; Kongtawelert, Prachya; Panthong, Ampai; Reutrakul, Vichai

2012-01-01

277

Bortezomib down-modulates the survival factor interferon regulatory factor 4 in Hodgkin lymphoma cell lines and decreases the protective activity of Hodgkin lymphoma-associated fibroblasts.  

Science.gov (United States)

Bortezomib is a proteasome inhibitor active in classical Hodgkin lymphoma (cHL) cell lines, but poorly active in the clinic when used as a single agent, suggesting that the microenvironment could protect from drug efficacy. Therefore, we investigated the effects of bortezomib activity in the presence of HL-associated fibroblasts (HL-AFs) and sCD40L. We found that co-cultivation with human HL-AFs or the addition of sCD40L during bortezomib treatment protected cHL cells from apoptosis and cytotoxicity and rescued the down-regulation of the survival factor interferon regulatory factor 4 (IRF4). In contrast, bortezomib treatment before co-cultivation with HL-AFs inhibited in a dose-dependent manner cHL cell adhesion to HL-AFs and completely overcame HL-AF protection against drug activity. Consistently, we found that bortezomib treatment down-regulated the surface expression of CD49d and CD44, which mediate the adhesion of cHL cells to HL-AFs, and of CD54 and CD40, which mediate the adhesion to CD40L+ rosetting T-cells. These preclinical findings suggest that the low in vivo activity of bortezomib as a single agent may be due to a protective influence of the microenvironment. However, inclusion of bortezomib in the cHL drug regimen, by reducing IRF4 expression and interactions with the microenvironment, could increase the efficacy of current chemotherapeutic treatment of relapsed/refractory cHL. PMID:23647062

Celegato, Marta; Borghese, Cinzia; Casagrande, Naike; Carbone, Antonino; Colombatti, Alfonso; Aldinucci, Donatella

2014-01-01

278

Granulocyte-macrophage colony-stimulating factor activates the transcription of nuclear factor kappa B and induces the expression of nitric oxide synthase in a skin dendritic cell line.  

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Nitric oxide (NO) produced by skin dendritic cells and keratinocytes plays an important role in skin physiology, growth and remodelling. Nitric oxide is also involved in skin inflammatory processes and in modulating antigen presentation (either enhancing or suppressing it). In this study, we found that GM-CSF stimulates the expression of the inducible isoform of nitric oxide synthase (iNOS) in a fetal-skin-derived dendritic cell line (FSDC) and, consequently, increases the nitrite production ...

Cruz, Mt; Duarte, Cb; Gonc?alo, Margarida; Figueiredo, A.; Carvalho, Ap; Lopes, Mc

2001-01-01

279

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  

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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...

2010-01-01

280

Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line  

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Sarcodiol (SD) is a semi-synthetic derivative of sarcophine, a marine natural product. In our previous work, we reported the significant chemopreventive effects of SD against non-melanoma skin cancer both in vitro and in vivo mouse models. In this investigation, we extended this work to study the effect of sarcodiol on melanoma development, the more deadly form of skin cancer, using the mouse melanoma B16F10 cell line. In this study we report that SD inhibits the de novo DNA synthesis and enh...

2011-01-01

 
 
 
 
281

Sarcophine-Diol, a Skin Cancer Chemopreventive Agent, Inhibits Proliferation and Stimulates Apoptosis in Mouse Melanoma B16F10 Cell Line  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Sarcodiol (SD) is a semi-synthetic derivative of sarcophine, a marine natural product. In our previous work, we reported the significant chemopreventive effects of SD against non-melanoma skin cancer both in vitro and in vivo mouse models. In this investigation, we extended this work to study the effect of sarcodiol on melanoma development, the more deadly form of skin cancer, using the mouse melanoma B16F10 cell line. In this study we report that SD inhibits the de novo DNA synthesis and enh...

2012-01-01

282

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

283

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

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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

284

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

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Two fibroblast cell strains derived from human embryonic lungs (WI-38 and IMR-90) were transformed into neoplastic cells by treatment with Co-60 ..gamma..-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 fibroblasts for transformation studies are discussed. Finally, future problems in transformation of human cells are described.

Namba, M.; Nishitani, K.; Fukushima, F.; Kimoto, T. (Kawasaki Medical School, Kurashiki, Okayama (Japan)); Utsunomiya, J.

1981-01-01

285

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 fibroblasts for transformation studies are discussed. Finally, future problems in transformation of human cells are described. (J.P.N.)

1981-01-01

286

Cholesteatoma Fibroblasts Promote Epithelial Cell Proliferation through Overexpression of Epiregulin  

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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

The effect of tissue expansion on dermal fibroblast contraction.  

Science.gov (United States)

Tissue expansion alters the function of skin cells. We studied the effects of expansion on the contractile function of dermal fibroblasts using an in vitro model, the fibroblast-populated collagen lattice (FPCL). Spherical expanders were placed dorsally in 30 Sprague-Dawley rats; one-half were serially inflated. One, 2, and 4 weeks later, 5 rats from each group were killed. Fibroblasts were cultured from dermis overlying the expanders and mixed with collagen, medium, and serum in petri dishes to form FPCL. Fibroblasts from 5 rats that had not undergone surgery were cultured to make control FPCL. Contraction was assessed by measuring the areas of the FPCL. At 48 hours, FPCL containing expanded fibroblasts had contracted significantly less than those containing unexpanded or control fibroblasts. Four weeks of expansion resulted in less contraction than 1 or 2 weeks. Tissue expansion inhibits the in vitro contractile function of dermal fibroblasts in the rat in a time-related fashion. PMID:1596063

Chang, B; Tuchler, R E; Siebert, J W; Longaker, M T; Burd, D A

1992-04-01

288

Skin graft  

Science.gov (United States)

Skin transplant; Skin autografting; FTSG; STSG; Split thickness skin graft; Full thickness skin graft ... site. Most people who are having a skin graft have a split-thickness skin graft. This takes ...

289

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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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 u...

Selvey Saxon; Haupt Larisa M; Thompson Erik W; Matthaei Klaus I; Irving Michael G; Griffiths Lyn R

2004-01-01

290

Induction of DNA protein cross-links in the C13-cell line of hamster fibroblasts by neutrons p(34)+Be  

International Nuclear Information System (INIS)

We use the method of alkaline elution according to K.W. Kohn et al. for the treatment of cell DNA in C13 cells (chinese hamster fibroblasts) which were irradiated with low doses of either ?-rays or neutrons. We observe the formation of covalent links between DNA and proteins. Neutrons are more efficient than ?-rays in producing these cross-links at low doses 0-6 Gy. These incubation of irradiated cells during 2 hours at 37 0C

1984-01-01

291

Parental somatic and germ-line mosaicism for a FBN2 mutation and analysis of FBN2 transcript levels in dermal fibroblasts.  

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Congenital contractural arachnodactyly (CCA) is an autosomal dominant disorder that is phenotypically related to the Marfan syndrome. CCA has recently been shown to result from mutations in the FBN2 gene, which encodes an elastin-associated microfibrillar protein called fibrillin-2. Two siblings are reported here with classic manifestations of CCA with unaffected parents. Analysis of the FBN2 cDNA from dermal fibroblasts from one of the affected siblings revealed a heterozygous exon splicing ...

1997-01-01

292

A comparative transmission electron microscopy study of titanium dioxide and carbon black nanoparticles uptake in human lung epithelial and fibroblast cell lines.  

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Several studies suggest that the biological responses induced by manufactured nanoparticles (MNPs) may be linked to their accumulation within cells. However, MNP internalisation has not yet been sufficiently characterised. Therefore, the aim of this study was to compare the intracellular uptake of three different MNPs: two made of carbon black (CB) and one made of titanium dioxide (TiO(2)), in 16HBE bronchial epithelial cells and MRC5 fibroblasts. Transmission electron microscopy was used to ...

Belade, Esther; Armand, Lucie; Martinon, Laurent; Kheuang, Laurence; Fleury-feith, Jocelyne; Baeza-squiban, Armelle; Lanone, Sophie; Billon-galland, Marie-annick; Pairon, Jean-claude; Boczkowski, Jorge

2012-01-01

293

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

Science.gov (United States)

Reprogramming somatic cells into induced pluripotent stem cells (iPSCs) has been suspected of causing de novo copy number variation. To explore this issue, here we perform a whole-genome and transcriptome analysis of 20 human iPSC lines derived from the primary skin fibroblasts of seven individuals using next-generation sequencing. We find that, on average, an iPSC line manifests two copy number variants (CNVs) not apparent in the fibroblasts from which the iPSC was derived. Using PCR and digital droplet PCR, we show that at least 50% of those CNVs are present as low-frequency somatic genomic variants in parental fibroblasts (that is, the fibroblasts from which each corresponding human iPSC line is derived), and are manifested in iPSC lines owing to their clonal origin. Hence, reprogramming does not necessarily lead to de novo CNVs in iPSCs, because most of the line-manifested CNVs reflect somatic mosaicism in the human skin. Moreover, our findings demonstrate that clonal expansion, and iPSC lines in particular, can be used as a discovery tool to reliably detect low-frequency CNVs in the tissue of origin. Overall, we estimate that approximately 30% of the fibroblast cells have somatic CNVs in their genomes, suggesting widespread somatic mosaicism in the human body. Our study paves the way to understanding the fundamental question of the extent to which cells of the human body normally acquire structural alterations in their DNA post-zygotically. PMID:23160490

Abyzov, Alexej; Mariani, Jessica; Palejev, Dean; Zhang, Ying; Haney, Michael Seamus; Tomasini, Livia; Ferrandino, Anthony F; Rosenberg Belmaker, Lior A; Szekely, Anna; Wilson, Michael; Kocabas, Arif; Calixto, Nathaniel E; Grigorenko, Elena L; Huttner, Anita; Chawarska, Katarzyna; Weissman, Sherman; Urban, Alexander Eckehart; Gerstein, Mark; Vaccarino, Flora M

2012-12-20

294

The ultrastructure and etiology of chronic radiotherapy damage in human skin  

International Nuclear Information System (INIS)

Ulcerated and nonulcerated skin from 5 patients with chronic radiation skin damage was examined using electron microscopy. Noticeable fibroblast disorganization was seen, with swollen and degenerating mitochondria, multiple vacuoles, and dilated irregular rough endoplasmic reticulum. Unusual crystalline inclusions were seen in some fibroblasts. In the ulcerated skin, contractile fibroblasts (myofibroblasts) were seen in 2 of 4 specimens. Stroma showed dense collagen and prominent elastosis. The microvasculature in the radiation-damaged tissue showed occasional lumen occlusion and vacuolization of endothelial cells, without consistent abnormality. These data suggest that permanent damage to fibroblasts or fibroblast stem cells may play an important role in chronic radiation skin ulceration

1982-01-01

295

Human fibroblast collagenase: glycosylation and tissue-specific levels of enzyme synthesis.  

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Human skin fibroblasts secrete collagenase as two proenzyme forms (57 and 52 kDa). The minor (57-kDa) proenzyme form is the result of a partial posttranslational modification of the major (52-kDa) proenzyme through the addition of N-linked complex oligosaccharides. Human endothelial cells as well as fibroblasts from human colon, cornea, gingiva, and lung also secrete collagenase in two forms indistinguishable from those of the skin fibroblast enzyme. In vitro tissue culture studies have shown...

Wilhelm, S. M.; Eisen, A. Z.; Teter, M.; Clark, S. D.; Kronberger, A.; Goldberg, G.

1986-01-01

296

Abnormal proliferation and aging of cultured fibroblasts from pigs with subcutaneous fibrosis induced by gamma irradiation  

International Nuclear Information System (INIS)

In vivo, fibrotic disorders, which may be due either to injury or disease, are characterized by overproliferation of fibroblasts and overproduction of connective tissue. In vitro, however, most of the fibrotic cell lines studied exhibited no differences in growth potential compared with control cell lines derived from normal skin. In the present study, we investigated the in vitro behavior of fibroblasts derived from pigs with subcutaneous fibrosis induced by gamma irradiation. The cells were isolated from the scar tissue six to 20 months after irradiation. In primary cultures, the cells derived from the fibrotic lesions exhibited greater attachment efficiency and faster proliferation than those of the cells derived from the normal skin of the same animal. In long-term cultures, the differences between normal and fibrotic cells were still greater: the normal skin cells underwent 17 population doublings and then died, whereas the fibrotic cells exhibited a prolonged life span, and were still actively proliferating after 80 population doublings. Cell morphology and the number of chromosomes were modified throughout subcultures. These results imply that in the scar tissue active fibrotic cell proliferation continued for years after irradiation and that this activation was expressed in vitro. Therefore, in this pig fibrosis model, the data acquired in the present in vitro studies closely resemble that obtained from earlier in vivo observations

1989-01-01

297

Examination of Early Interactions between Haemophilus ducreyi and Host Cells by Using Cocultured HaCaT Keratinocytes and Foreskin Fibroblasts  

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Haemophilus ducreyi is the etiologic agent of chancroid, a sexually transmitted genital ulcer disease. Keratinocytes are likely the first cell type encountered by H. ducreyi upon infection of human skin; thus, the interaction between H. ducreyi and keratinocytes is probably important for the ability of H. ducreyi to establish infection. We have used the HaCaT keratinocyte cell line grown in monolayers and in cocultures with HS27 fibroblasts to investigate H. ducreyi interactions with keratino...

Zaretzky, Franca R.; Kawula, Thomas H.

1999-01-01

298

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

Science.gov (United States)

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 droplet PCR (ddPCR), we show that at least 50% of those CNVs are present as low frequency somatic genomic variants in parental fibroblasts (i.e. the fibroblasts from which each corresponding hiPSC line is derived) and are manifested in iPSC colonies due to the colonies’ clonal origin. Hence, reprogramming does not necessarily lead to de novo CNVs in iPSC, since most of line-manifested CNVs reflect somatic mosaicism in the human skin. Moreover, our findings demonstrate that clonal expansion, and iPSC lines in particular, can be used as a discovery tool to reliably detect low frequency CNVs in the tissue of origin. Overall, we estimate that approximately 30% of the fibroblast cells have somatic CNVs in their genomes, suggesting widespread somatic mosaicism in the human body. Our study paves the way to understanding the fundamental question of the extent to which cells of the human body normally acquire structural alterations in their DNA post-zygotically.

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; Weissman, Sherman; Urban, Alexander Eckehart; Gerstein, Mark; Vaccarino, Flora M.

2012-01-01

299

Parental somatic and germ-line mosaicism for a FBN2 mutation and analysis of FBN2 transcript levels in dermal fibroblasts.  

Science.gov (United States)

Congenital contractural arachnodactyly (CCA) is an autosomal dominant disorder that is phenotypically related to the Marfan syndrome. CCA has recently been shown to result from mutations in the FBN2 gene, which encodes an elastin-associated microfibrillar protein called fibrillin-2. Two siblings are reported here with classic manifestations of CCA with unaffected parents. Analysis of the FBN2 cDNA from dermal fibroblasts from one of the affected siblings revealed a heterozygous exon splicing error deleting nt 3722-3844 of the FBN2 mRNA. This cDNA deletion resulted in selective removal of one of the 43 calcium-binding EGF-like domains of the fibrillin-2 protein. Analysis of the FBN2 gene in the affected siblings' DNA indicated that the splicing error resulted from an A-to-G transition 15 nt upstream from the 3' splice site of the intron. The genomic mutation resulting in the splicing error alters a putative branch point sequence important for lariat formation, an intermediate structure of normal splicing. The mutation was detectable in DNA from the father's hair bulbs and buccal cells but not his white blood cell DNA, indicating that the father was a somatic mosaic. Analysis of transcript levels by use of dermal fibroblasts from the proband demonstrated that the FBN2 allele containing the exon deletion was expressed at a higher level than the allele inherited from the mother. These results indicate that FBN2 exon splicing errors are a cause of CCA, furthering the understanding of the molecular basis of this disorder. In addition, the demonstration of gonadal mosaicism in the FBN2 gene is important for accurate genetic counseling of families with sporadic cases of CCA. Finally, the preferential expression of the mutated FBN2 allele in dermal fibroblasts may have implications for understanding the pathogenesis and rarity of CCA. PMID:9106527

Putnam, E A; Park, E S; Aalfs, C M; Hennekam, R C; Milewicz, D M

1997-04-01

300

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 ...

 
 
 
 
301

Radioprotection by glutathione ester: transport of glutathione ester into human lymphoid cells and fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

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 exceed those found normally. Glutathione ester was found to protect human lymphoid cells of the CEM line against the lethal effects of irradiation. Under the conditions employed, complete protection was found when the ester was added prior to irradiation. Addition of the ester after irradiation was partially effective, suggesting that GSH may also function in repair processes. 20 references, 3 figures, 1 table.

Wellner, V.P.; Anderson, M.E.; Puri, R.N.; Jensen, G.L.; Meister, A.

1984-08-01

302

Small interfering RNA mediated Poly (ADP-ribose Polymerase-1 inhibition upregulates the heat shock response in a murine fibroblast cell line  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Poly (ADP-ribose polymerase-1 (PARP-1 is a highly conserved multifunctional enzyme, and its catalytic activity is stimulated by DNA breaks. The activation of PARP-1 and subsequent depletion of nicotinamide adenine dinucleotide (NAD+ and adenosine triphosphate (ATP contributes to significant cytotoxicity in inflammation of various etiologies. On the contrary, induction of heat shock response and production of heat shock protein 70 (HSP-70 is a cytoprotective defense mechanism in inflammation. Recent data suggests that PARP-1 modulates the expression of a number of cellular proteins at the transcriptional level. In this study, small interfering RNA (siRNA mediated PARP-1 knockdown in murine wild-type fibroblasts augmented heat shock response as compared to untreated cells (as evaluated by quantitative analysis of HSP-70 mRNA and HSP-70 protein expression. These events were associated with increased DNA binding of the heat shock factor-1 (HSF-1, the major transcription factor of the heat shock response. Co-immunoprecipitation experiments in nuclear extracts of the wild type cells demonstrated that PARP-1directly interacted with HSF-1. These data demonstrate that, in wild type fibroblasts, PARP-1 plays a pivotal role in modulating the heat shock response both through direct interaction with HSF-1 and poly (ADP-ribosylation.

Zingarelli Basilia

2011-02-01

303

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

304

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

305

On-line monitoring of UV-induced lipid peroxidation products from human skin in vivo using proton-transfer reaction mass spectrometry  

Science.gov (United States)

Proton-transfer reaction mass spectrometry (PTR-MS) was used to study ultraviolet (UV) light-induced lipid peroxidation in human skin, in vivo. Emissions of volatile organic compounds (VOCs) in the mass range between 20 and 150 amu in the headspace of the skin of 16 healthy volunteers were monitored before, during and after irradiation in an on-line and non-invasive fashion. From these experiments, five volatile substances were found to reflect the damage caused by UV-radiation. The two major compounds (monitored at mass 45 and 59 amu) were identified as acetaldehyde and propanal using a combination of Tenax-based gas chromatographic pre-separation with PTR-MS. The other volatiles (with characteristic ions at, among others, masses 73 and 87 amu) could not be identified. Simultaneous measurement of the established lipid peroxidation biomarker ethene using laser-based photoacoustic trace gas detection revealed a similar pattern and statistically significant correlations between VOC production measured with PTR-MS and ethene. Variations in UV-radiation intensity were reflected by the amount of acetaldehyde and propanal emitted from the skin. Our results show that acetaldehyde and propanal can be used as biomarkers for lipid peroxidation.

Steeghs, Marco M. L.; Moeskops, Bas W. M.; van Swam, Karen; Cristescu, Simona M.; Scheepers, Paul T. J.; Harren, Frans J. M.

2006-06-01

306

Skin toxicity and quality of life in patients with metastatic colorectal cancer during first-line panitumumab plus FOLFIRI treatment in a single-arm phase II study  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Integument-related toxicities are common during epidermal growth factor receptor (EGFR-targeted therapy. Panitumumab is a fully human monoclonal antibody targeting the EGFR that significantly improves progression-free survival when added to chemotherapy in patients with metastatic colorectal cancer who have wild-type (WT KRAS tumours. Primary efficacy and tolerability results from a phase II single-arm study of first-line panitumumab plus FOLFIRI in patients with metastatic colorectal cancer have been reported. Here we report additional descriptive tolerability and quality of life data from this trial. Methods Integument-related toxicities and quality of life were analysed; toxicities were graded using modified National Cancer Institute Common Toxicity Criteria. Kaplan-Meier estimates of time to and duration of first integument-related toxicity were prepared. Quality of life was measured using EuroQoL EQ-5D and EORTC QLQ-C30. Best overall response was analysed by skin toxicity grade and baseline quality of life. Change in quality of life was analysed by skin toxicity severity. Results 154 patients were enrolled (WT KRAS n?=?86; mutant KRAS n?=?59; most (98% experienced integument-related toxicities (most commonly rash [42%], dry skin [40%] and acne [36%]. Median time to first integument-related toxicity was 8 days; median duration was 334 days. Overall, proportionally more patients with grade 2+ skin toxicity responded (56% compared with those with grade 0/1 (29%. Mean overall EQ-5D health state index scores (0.81 vs. 0.78, health rating scores (72.5 vs. 71.0 and QLQ-C30 global health status scores (65.8 vs. 66.7 were comparable at baseline vs. safety follow-up (8 weeks after completion, respectively and appeared unaffected by skin toxicity severity. Conclusions First-line panitumumab plus FOLFIRI has acceptable tolerability and appears to have little impact on quality of life, despite the high incidence of integument-related toxicity. Trial registration ClinicalTrials.gov NCT00508404

Thaler Josef

2012-09-01

307

Regulated Proenkephalin Expression in Human Skin and Cultured Skin Cells  

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Skin responds to environmental stressors via coordinated actions of the local neuroimmunoendocrine system. Although some of these responses involve opioid receptors, little is known about cutaneous proenkephalin expression, its environmental regulation, and alterations in pathology. The objective of this study was to assess regulated expression of proenkephalin in normal and pathological skin and in isolated melanocytes, keratinocytes, fibroblasts, and melanoma cells. The proenkephalin gene a...

Slominski, Andrzej T.; Zmijewski, Michal A.; Zbytek, Blazej; Brozyna, Anna A.; Granese, Jackie; Pisarchik, Alexander; Szczesniewski, Andre; Tobin, Desmond J.

2011-01-01

308

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.)

1980-01-01

309

Positive correlation between the efficiency of induced pluripotent stem cells and the development rate of nuclear transfer embryos when the same porcine embryonic fibroblast lines are used as donor cells.  

Science.gov (United States)

Induced pluripotent stem cells (iPSCs) and nuclear transfer (NT) are two of the primary routes to reprogram differentiated cells back to the pluripotent state. However, it is still unknown whether there is any correlation between the reprogramming efficiency of iPSCs and NT if the same donor cells are employed. In this study, six porcine embryonic fibroblast (PEF) lines from Landrace (L1, L6, L9) or Congjiang local pigs (C4, C5, C6) were used for iPSC induction and NT. Furthermore, the resultant iPSCs from four PEF lines (L1, L6, C4, and C5) were used for NT (iPSC-NT), and the expression of exogenous genes was detected in iPSC-NT embryos by real-time PCR. The results showed that the efficiency of iPSC lines established from different PEF lines were significantly different. When the same PEF lines were used as donor cells for NT, the blastocysts rates were also different among different PEF lines and positively related with iPSCs induction efficiency. When the iPSCs were used as donor cells for NT, compared with the source PEFs, the blastocysts rates were significantly decreased. Real-time PCR results indicated that exogenous genes (Oct4, c-Myc) continued to be expressed in iPSC-NT embryos. In summary, our results demonstrate that there was a positive correlation between iPSCs and NT reprogramming efficiency, although the mechanism of these two routes is different. This may provide a new method to select the appropriate donor cells for inducing iPSCs. PMID:24738969

Xie, Bingteng; Wang, Jianyu; Liu, Shichao; Wang, Jiaqiang; Xue, Binghua; Li, Jingyu; Wei, Renyue; Zhao, Yanhua; Liu, Zhonghua

2014-06-01

310

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

311

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

2014-01-03

312

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 ...

313

Human skin is a steroidogenic tissue: steroidogenic enzymes and cofactors are expressed in epidermis, normal sebocytes, and an immortalized sebocyte cell line (SEB-1).  

Science.gov (United States)

Although the human sebaceous gland can synthesize cholesterol from acetate and can further metabolize steroids such as dehydroepiandrosterone into potent androgens, the de novo production of steroids from cholesterol has not been demonstrated in human skin. The goal of this study was to delineate the steroidogenic pathway upstream from dehydroepiandrosterone by documenting the presence of members of the P450 side chain cleavage system (P450scc). This system catalyzes the initial step in steroid hormone synthesis following translocation of cholesterol to the inner mitochondrial membrane. In concert with its cofactors, adrenodoxin and adrenodoxin reductase, and the transcription factor steroidogenic factor 1, P450scc converts cholesterol to pregnenolone. An SV40 immortalized human sebaceous gland cell line (SEB-1) was established in order to facilitate investigation of the P450scc system. The sebaceous phenotype of SEB-1 sebocytes was confirmed using immunohistochemistry, Oil Red O staining, and gene array expression analysis. Presence of P450scc, adrenodoxin reductase, cytochrome P450 17-hydroxylase (P450c17), and steroidogenic factor 1 was documented in human facial skin, human sebocytes, and SEB-1 sebocytes. Using immunohistochemistry, antibodies to the above proteins localized to epidermis, hair follicles, sebaceous ducts, and sebaceous glands in sections of facial skin. Results of immunohistochemistry were confirmed with Western blotting. Biochemical activity of cytochrome P450scc and P450c17 was demonstrated in SEB-1 sebocytes using radioimmunoassay. The relative abundance of mRNA for P450scc, P450c17, and steroidogenic factor 1 in SEB-1 sebocytes and sebaceous glands was compared to mRNA levels in ovarian theca and granulosa cells using real-time quantitative polymerase chain reaction. Gene array expression analysis and quantitative polymerase chain reaction indicated that mRNA for P450scc is more abundant than mRNA for both P450c17 and steroidogenic factor 1 in sebaceous glands and SEB-1 cells. These data demonstrate that the skin is in fact a steroidogenic tissue. The clinical significance of this finding in mediating androgenic skin disorders such as acne, hirsutism, or androgenetic alopecia remains to be established. PMID:12787114

Thiboutot, Diane; Jabara, Sami; McAllister, Jan M; Sivarajah, Aruntha; Gilliland, Kathyrn; Cong, Zhaoyuan; Clawson, Gary

2003-06-01

314

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.  

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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

315

Fetal ACL Fibroblasts Exhibit Enhanced Cellular Properties Compared with Adults  

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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

316

Overexpression of the IGF-II/M6P receptor in mouse fibroblast cell lines differentially alters expression profiles of genes involved in Alzheimer's disease-related pathology.  

Science.gov (United States)

Alzheimer's disease (AD) is the most common type of senile dementia affecting elderly people. The processing of amyloid precursor protein (APP) leading to the generation of ?-amyloid (A?) peptide contributes to neurodegeneration and development of AD pathology. The endocytic trafficking pathway, which comprises of the endosomes and lysosomes, acts as an important site for A? generation, and endocytic dysfunction has been linked to increased A? production and loss of neurons in AD brains. Since insulin-like growth factor-II (IGF-II) receptor plays a critical role in the transport of lysosomal enzymes from the trans-Golgi network to endosomes, it is likely that the receptor may have a role in regulating A? metabolism in AD pathology. However, very little is known on how altered levels of the IGF-II receptor can influence the expression/function of various molecules involved in AD pathology. To address this issue, we evaluated the expression profiles of 87 selected genes related to AD pathology in mouse fibroblast MS cells that are deficient in murine IGF-II receptor and corresponding MS9II cells overexpressing ? 500 times the human IGF-II receptors. Our results reveal that an elevation in IGF-II receptor levels alters the expression profiles of a number of genes including APP as well as enzymes regulating A? production, degradation and clearance mechanisms. Additionally, it influences the expression of various lysosomal enzymes and protein kinases that are involved in A? toxicity. IGF-II receptor overexpression also alters expression of several genes involved in intracellular signalling as well as cholesterol metabolism, which play a critical role in AD pathology. The altered gene profiles observed in this study closely match with the corresponding protein levels, with a few exceptions. These results, taken together, suggest that an elevation in IGF-II receptor levels can influence the expression profiles of transcripts as well as proteins that are involved in AD pathogenesis. PMID:24846272

Wang, Yanlin; Thinakaran, Gopal; Kar, Satyabrata

2014-01-01

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 10"6 U937 cells/ml of serum-free RPMI medium were incubated with 100 ng/ml of phorbol myristate acetate for 48 hr at 37"0C in 5% CO_2 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

1986-03-05

318

Overexpression of the IGF-II/M6P Receptor in Mouse Fibroblast Cell Lines Differentially Alters Expression Profiles of Genes Involved in Alzheimer's Disease-Related Pathology  

Science.gov (United States)

Alzheimer’s disease (AD) is the most common type of senile dementia affecting elderly people. The processing of amyloid precursor protein (APP) leading to the generation of ?-amyloid (A?) peptide contributes to neurodegeneration and development of AD pathology. The endocytic trafficking pathway, which comprises of the endosomes and lysosomes, acts as an important site for A? generation, and endocytic dysfunction has been linked to increased A? production and loss of neurons in AD brains. Since insulin-like growth factor-II (IGF-II) receptor plays a critical role in the transport of lysosomal enzymes from the trans-Golgi network to endosomes, it is likely that the receptor may have a role in regulating A? metabolism in AD pathology. However, very little is known on how altered levels of the IGF-II receptor can influence the expression/function of various molecules involved in AD pathology. To address this issue, we evaluated the expression profiles of 87 selected genes related to AD pathology in mouse fibroblast MS cells that are deficient in murine IGF-II receptor and corresponding MS9II cells overexpressing ?500 times the human IGF-II receptors. Our results reveal that an elevation in IGF-II receptor levels alters the expression profiles of a number of genes including APP as well as enzymes regulating A? production, degradation and clearance mechanisms. Additionally, it influences the expression of various lysosomal enzymes and protein kinases that are involved in A? toxicity. IGF-II receptor overexpression also alters expression of several genes involved in intracellular signalling as well as cholesterol metabolism, which play a critical role in AD pathology. The altered gene profiles observed in this study closely match with the corresponding protein levels, with a few exceptions. These results, taken together, suggest that an elevation in IGF-II receptor levels can influence the expression profiles of transcripts as well as proteins that are involved in AD pathogenesis.

Wang, Yanlin; Thinakaran, Gopal; Kar, Satyabrata

2014-01-01

319

Replacement of murine fibroblasts by human fibroblasts irradiated in obtaining feeder layer for the culture of human keratinocytes  

International Nuclear Information System (INIS)

Human autologous epithelia cultivated in vitro, have been used successfully in treating damage to skin integrity. The methodology allowed the cultivation of these epithelia was described by Rheinwald and Green in 1975, this methodology consisted in seeding keratinocytes onto a feeder layer composed of lineage 3T3 murine fibroblasts, the proliferation rate is controlled through the action of ionizing radiation. However, currently there is a growing concern about the possibility of transmitting prions and murine viruses to transplanted patients. Taking into account this concern, in this present work, we replaced the feeder layer originally composed of murine fibroblasts by human fibroblasts. To obtain this new feeder layer was necessary to standardize the enough irradiation dose to inhibit the replication of human fibroblasts and the verification of effectiveness of the development of keratinocytes culture on a feeder layer thus obtained. According to the obtained results we can verify that the human fibroblasts irradiated at various tested doses (60, 70, 100, 200, 250 and 300 Gy) had their mitotic activity inactivated by irradiation, allowing the use of any of these doses to confection of the feeder layer, since these fibroblasts irradiated still showed viable until fourteen days of cultivation. In the test of colony formation efficiency was observed that keratinocytes seeded on irradiated human fibroblasts were able to develop satisfactorily, preserving their clonogenic potential. Therefore it was possible the replacement of murine fibroblasts by human fibroblasts in confection of the feeder layer, in order to eliminate this xenobiotic component of the keratinocytes culture. (author)

2011-10-24

320

Replacement of murine fibroblasts by human fibroblasts irradiated in obtaining feeder layer for the culture of human keratinocytes  

Energy Technology Data Exchange (ETDEWEB)

Human autologous epithelia cultivated in vitro, have been used successfully in treating damage to skin integrity. The methodology allowed the cultivation of these epithelia was described by Rheinwald and Green in 1975, this methodology consisted in seeding keratinocytes onto a feeder layer composed of lineage 3T3 murine fibroblasts, the proliferation rate is controlled through the action of ionizing radiation. However, currently there is a growing concern about the possibility of transmitting prions and murine viruses to transplanted patients. Taking into account this concern, in this present work, we replaced the feeder layer originally composed of murine fibroblasts by human fibroblasts. To obtain this new feeder layer was necessary to standardize the enough irradiation dose to inhibit the replication of human fibroblasts and the verification of effectiveness of the development of keratinocytes culture on a feeder layer thus obtained. According to the obtained results we can verify that the human fibroblasts irradiated at various tested doses (60, 70, 100, 200, 250 and 300 Gy) had their mitotic activity inactivated by irradiation, allowing the use of any of these doses to confection of the feeder layer, since these fibroblasts irradiated still showed viable until fourteen days of cultivation. In the test of colony formation efficiency was observed that keratinocytes seeded on irradiated human fibroblasts were able to develop satisfactorily, preserving their clonogenic potential. Therefore it was possible the replacement of murine fibroblasts by human fibroblasts in confection of the feeder layer, in order to eliminate this xenobiotic component of the keratinocytes culture. (author)

Yoshito, Daniele; Sufi, Bianca S.; Santin, Stefany P.; Mathor, Monica B. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Altran, Silvana C.; Isaac, Cesar [Universidade Sao Paulo (USP), Sao Paulo, SP (Brazil). Fac. de Medicina. Lab. de Microcirurgia Plastica; Esteves-Pedro, Natalia M. [Universidade Sao Paulo (USP), Sao Paulo, SP (Brazil). Fac. de Ciencias Farmaceuticas. Lab. de Controle Biologico; Herson, Marisa R. [DonorTissue Bank of Victoria (Australia)

2011-07-01

 
 
 
 
321

Solar aging of the skin  

Energy Technology Data Exchange (ETDEWEB)

Aging of the skin is a combination of chronological aging and degenerative influences from the environment. Most important is the influence of ultraviolet radiation upon dermal fibroblasts. As a result of changes in collagen and elastic fibres with deposition of ''solar elastosis'', chronically sun-exposed skin looses elasticity and develops wrinkles. Results of recent investigations show that solar degenerative changes of the skin are spontaneously repaired when the radiation is stopped and/or sun filters are used. This repair process is accelerated by the topical use of retinoic acid. 30 refs.

Fyrand, O. (Rikshospitalet, Oslo (Norway))

1989-03-01

322

Solar aging of the skin  

International Nuclear Information System (INIS)

Aging of the skin is a combination of chronological aging and degenerative influences from the environment. Most important is the influence of ultraviolet radiation upon dermal fibroblasts. As a result of changes in collagen and elastic fibres with deposition of ''solar elastosis'', chronically sun-exposed skin looses elasticity and develops wrinkles. Results of recent investigations show that solar degenerative changes of the skin are spontaneously repaired when the radiation is stopped and/or sun filters are used. This repair process is accelerated by the topical use of retinoic acid

1989-03-01

323

Effect of Gly-Gly-His, Gly-His-Lys and their copper complexes on TNF-alpha-dependent IL-6 secretion in normal human dermal fibroblasts.  

Science.gov (United States)

Cosmeceuticals represent a marriage between cosmetics and pharmaceuticals. There are numerous cosmeceutically active products which can be broadly classified into the following categories: antioxidants, oligopeptides, growth factors and pigment lightning agents. Much attention has been focused on the tripeptides such as Gly-His-Lys (GHK) and Gly-Gly-His (GGH) and their copper complexes, which have a high activity and good skin tolerance. Recent data suggested their physiological role in process of wound healing, tissue repair and skin inflammation. The mechanism of anti-inflammatory properties of these peptides is not clear. The aim of the study was evaluation of influence of two peptides GGH. GHK and their copper complexes and saccharomyces/copper ferment (Oligolides Copper) on secretion of pro-inflammatory IL-6 in normal human dermal fibroblasts NHDF cell line. IL-6 was evaluated using the ELISA kit. GGH, GHK, CuCl2 and their copper complexes decreased TNF-alpha-dependent IL-6 secretion in fibroblasts. IL-6 is crucial for normal wound healing, skin inflammation and UVB-induced erythema. Because of the anti-inflammatory properties, the copper-peptides could be used on the skin surface instead of corticosteroids or non-steroidal anti-inflammatory drugs, which have more side effects. Our observations provide some new information about the role of these tripeptides in skin inflammation. PMID:23285694

Gruchlik, Arkadiusz; Jurzak, Magdalena; Chodurek, Ewa; Dzierzewicz, Zofia

2012-01-01

324

The development of a 3D immunocompetent model of human skin.  

Science.gov (United States)

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 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. PMID:23880658

Chau, David Y S; Johnson, Claire; MacNeil, Sheila; Haycock, John W; Ghaemmaghami, Amir M

2013-09-01

325

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

326

Chronic ?-irradiation results in increased cell killing and chromosomal aberration with specific breakpoints in fibroblast cell strains derived from non-Hodgkin's lymphoma patients  

International Nuclear Information System (INIS)

Cultured skin fibroblast cells from 16 NHL (non-Hodgkin's lymphoma) patients and 2 clinically normal subjects were compared for cell survival and chromosomal aberration after chronic ?-irradiation. Fibroblasts from an ataxia telangiectasia (AT) homozygote and an AT heterozygote were used as positive controls. Following irradiation, fibroblasts from all 16 NHL patients showed an increase in both cell death and chromosomal aberration (breaks and rearrangements) compared to normal subjects. The difference in frequency of chromosomal aberration between normals and NHL-patients remained virtually unchanged over a period of 24-72 h post irradiation incubation of cells. Cell cycle analysis by flow cytometry carried out in 1 normal and 1 NHL fibroblast cell strain showed that more cells representing the NHL patient were in G2/M phase compared to the normal at various times of cytogenetic analysis. While the AT homozygote appeared to be the most radiosensitive, the AT heterozygote showed a slightly higher incidence of cell death and chromosomal aberration than the normals. The cellular and chromosomal radiosensitivity of fibroblast cell lines from NHL-patients differed slightly from that of AT heterozygote but clearly occupied an intermediate position between the AT homozygote and the normal subjects. Cells from 3 of the NHL patients showed radiation-induced specific chromosomal breaks involving chromosomes 1, 2, 6, 8, 10 and 11 which correspond to known fragile sites. Such breakpoints associated with increased radiosensitivity may be indicative of predisposition to malignancy in the patients studied. (author). 30 refs., 2 figs., 4 tabs

1992-12-16

327

Study of Fibroblast and Platelet Adhesion on Multi―wallCarbon Nanotubes  

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Abstract:Multi―walled nanotubes (MWCNTs) were grown on carbonpapers by chemical vapor deposition (CVD). Adhesion and proliferation of L929 mouse fibroblasts and plateletson MWCNTs and carbon papers were investigated. The influence of different bloodprotein adsorption on the human skin fibroblasts growth was also studied. The resultsshowed that mouse fibroblasts implanted on MWCNTs tended to grow more prolificallythan those on carbon papers. The cell concentration ob...

Zhao Meng-li, Yue Yu-chen

2010-01-01

328

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, ...

329

Chaperonin containing T-complex polypeptide (CCT) subunit expression in oral mucosal wounds and fibroblasts  

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Mucosal wound healing in adults has been reported to feature diminished scar formation compared to healing skin wounds. We sought to determine if the expression pattern of chaperonin containing T-complex polypeptide (CCT) subunits in mucosal wounds and fibroblasts is different from that observed in skin wounds and fibroblasts. We found that CCT-beta is the only subunit message to be reduced in wounded mucosa versus unwounded control, and this reduction was confirmed at the protein level. In c...

Satish, Latha; Lo, Nancy; Gallo, Phillip H.; Johnson, Sandra; Haberman, Stephanie; Kathju, Sandeep

2011-01-01

330

Comparative studies of collagen lattice contraction utilizing a normal and a transformed cell line.  

Science.gov (United States)

Differences between the behavior of cultured rat skin fibroblasts and that of a line of transformed rat sarcoma cells incorporated into a polymerized collagen lattice were examined. Fibroblast-populated collagen lattices (FPCL) were manufactured. Within 24 to 48 hr after manufacture, both cell lines reduced lattice size by a process known as lattice contraction. Contraction occurred more rapidly in both cell lines when the media were supplemented with 25% serum rather than the usual concentration of 10% serum. Similar growth patterns were observed with transformed cells within collagen lattices and on plastic surfaces. Normal rat fibroblasts were found to contract lattices faster than transformed cells. At the end of a 2-week period, the final contracted size of the transformed cell lattice was the same as that of normal cell lattices. The cellular density of transformed cells within the FPCL was eight times greater than that of FPCL made with normal rat cells. Normal rat fibroblasts elongated and flattened more, and organized the collagen matrix to a greater degree, than did transformed cells. In this instance, therefore, lattice contraction was shown to be linked more to the process of fibroblast elongation and collagen fiber organization than to cell number or density. PMID:6863399

Buttle, D J; Ehrlich, H P

1983-08-01

331

Overexpression of chicken interferon regulatory factor-1 (Ch-IRF-1) induces constitutive expression of MHC class I antigens but does not confer virus resistance to a permanent chicken fibroblast cell line.  

Science.gov (United States)

The chicken fibroblast cell line C32 has been transfected with the chicken homolog (Ch-IRF-1) of the mammalian transcription factor IRF-1. Stable transfectants were generated, constitutively overexpressing Ch-IRF-1 mRNA and protein. Cells overexpressing Ch-IRF-1 showed enhanced constitutive expression of MHC class I (B-F, beta-microglobulin) antigens. With increasing number of passages cells with normal B-F IV surface antigen expression accumulated. In the revertants, the amount of Ch-IRF-1 mRNA was reduced. Overexpression of Ch-IRF-1 had no effect on the constitutive expression and the induction by chicken interferon type-I and type-II (Ch-IFN) of guanylate-binding protein (GBP). Susceptibility to vesicular stomatitis virus, sindbis virus, Newcastle disease virus and vaccinia virus was not altered by overexpression of Ch-IRF-1. An antiviral state could be induced against all viruses tested by similar amounts of Ch-IFN type I in clone 20-18 expressing Ch-IRF-1 and cells transfected with empty vector. PMID:9831662

Zoeller, B; Popp, M; Walter, A; Redmann-Müller, I; Lodemann, E; Jungwirth, C

1998-11-19

332

Differential expression of matrix metalloproteases in human fibroblasts with different origins.  

Science.gov (United States)

Fibroblasts are widely distributed cells and are responsible for the deposition of extracellular matrix (ECM) components but also secrete ECM-degrading matrix metalloproteases. A finely balanced equilibrium between deposition and degradation of ECM is essential for structural integrity of tissues. In the past, fibroblasts have typically been understood as a uniform cell population with comparable functions regardless of their origin. Here, we determined growth curves of fibroblasts derived from heart, skin, and lung and clearly show the lowest proliferation rate for cardiac fibroblasts. Furthermore, we examined basal expression levels of collagen and different MMPs in these three types of fibroblasts and compared these concerning their site of origin. Interestingly, we found major differences in basal mRNA expression especially for MMP1 and MMP3. Moreover, we treated fibroblasts with TNF-? and observed different alterations under these proinflammatory conditions. In conclusion, fibroblasts show different properties in proliferation and MMP expression regarding their originated tissue. PMID:22500233

Lindner, Diana; Zietsch, Christin; Becher, P Moritz; Schulze, Karsten; Schultheiss, Heinz-Peter; Tschöpe, Carsten; Westermann, Dirk

2012-01-01

333

TNF-? Sensitizes Normal and Fibrotic Human Lung Fibroblasts to Fas-Induced Apoptosis  

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Pulmonary accumulation of fibroblasts and myofibroblasts in idiopathic pulmonary fibrosis/usual interstitial pneumonia (IFP/UIP) has been linked to (1) increased migration of a circulating pool of fibrocytes, (2) cell proliferation, and (3) resistance to apoptosis. The mechanism of physiologic apoptosis of lung fibroblasts is poorly understood. Using normal and fibrotic human lung fibroblasts and the human lung fibroblast cell line, MRC-5, we examined the regulation of Fas-induced apoptosis b...

Frankel, Stephen K.; Cosgrove, Gregory P.; Cha, Seung-ick; Cool, Carlyne D.; Wynes, Murry W.; Edelman, Benjamin L.; Brown, Kevin K.; Riches, David W. H.

2006-01-01

334

Establishment and transformation diminish the ability of fibroblasts to contract a native collagen gel  

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Cultures of established and transformed fibroblasts were less able to contract a hydrated collagen gel than normal precrisis cells. Postcrisis fibroblasts from different rodent strains and species underwent a further reduction in contraction ability and either spontaneous or simian virus 40 (SV40) transformation. Human precrisis fibroblasts contracted much more efficiently than two SV40-transformed human lines. Fibroblasts from a patient with Glanzmann's thrombasthenia were intermediate betwe...

1980-01-01

335

Cytokine Responses to Treponema pectinovorum and Treponema denticola in Human Gingival Fibroblasts  

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Human gingival fibroblasts were challenged with Treponema pectinovorum and Treponema denticola to test three specific hypotheses: (i) these treponemes induce different cytokine profiles from the fibroblasts, (ii) differences in cytokine profiles are observed after challenge with live versus killed treponemes, and (iii) differences in cytokine profiles are noted from different gingival fibroblast cell lines when challenged with these treponemes. Three normal gingival fibroblast cell cultures w...

Nixon, Connie S.; Steffen, Michelle J.; Ebersole, Jeffrey L.

2000-01-01

336

Grafting of Venous Leg Ulcers: An Ultra-Individual Comparison between Cultured Skin Equivalents and Full Thickness Skin Punches.  

Science.gov (United States)

Skin equivalents, consisting of a non-contracted collagen gel populated with allogeneic fibroblasts and covered with autologous cultured keratinocytes were used for grafting of venous leg ulcers. The results were compared in the same patient with those ob...

M. A. E. Mol B. P. Nanninga J. P. van Eendenburg W. Westerhof C. J. W. van Ginkel

1989-01-01

337

Resident fibroblast lineages mediate pressure overload-induced cardiac fibrosis  

Science.gov (United States)

Activation and accumulation of cardiac fibroblasts, which result in excessive extracellular matrix deposition and consequent mechanical stiffness, myocyte uncoupling, and ischemia, are key contributors to heart failure progression. Recently, endothelial-to-mesenchymal transition (EndoMT) and the recruitment of circulating hematopoietic progenitors to the heart have been reported to generate substantial numbers of cardiac fibroblasts in response to pressure overload–induced injury; therefore, these processes are widely considered to be promising therapeutic targets. Here, using multiple independent murine Cre lines and a collagen1a1-GFP fusion reporter, which specifically labels fibroblasts, we found that following pressure overload, fibroblasts were not derived from hematopoietic cells, EndoMT, or epicardial epithelial-to-mesenchymal transition. Instead, pressure overload promoted comparable proliferation and activation of two resident fibroblast lineages, including a previously described epicardial population and a population of endothelial origin. Together, these data present a paradigm for the origins of cardiac fibroblasts during development and in fibrosis. Furthermore, these data indicate that therapeutic strategies for reducing pathogenic cardiac fibroblasts should shift from targeting presumptive EndoMT or infiltrating hematopoietically derived fibroblasts, toward common pathways upregulated in two endogenous fibroblast populations.

Moore-Morris, Thomas; Guimaraes-Camboa, Nuno; Banerjee, Indroneal; Zambon, Alexander C.; Kisseleva, Tatiana; Velayoudon, Aurelie; Stallcup, William B.; Gu, Yusu; Dalton, Nancy D.; Cedenilla, Marta; Gomez-Amaro, Rafael; Zhou, Bin; Brenner, David A.; Peterson, Kirk L.; Chen, Ju; Evans, Sylvia M.

2014-01-01

338

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 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

1990-09-15

339

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

Energy Technology Data Exchange (ETDEWEB)

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 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.

Gilead, L.; Bibi, O.; Razin, E. (Hebrew Univ.-Hadassah Medical School, Jerusalem (Israel))

1990-09-15

340

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

 
 
 
 
341

Abscisic acid ameliorates the systemic sclerosis fibroblast phenotype in vitro  

Energy Technology Data Exchange (ETDEWEB)

Highlights: Black-Right-Pointing-Pointer ABA is an endogenous hormone in humans, regulating different cell responses. Black-Right-Pointing-Pointer ABA reverts some of the functions altered in SSc fibroblasts to a normal phenotype. Black-Right-Pointing-Pointer UV-B irradiation increases ABA content in SSc cultures. Black-Right-Pointing-Pointer 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-{beta} (TGF-{beta}). Conversely, migration toward ABA, but not toward TGF-{beta}, 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.

Bruzzone, Santina, E-mail: santina.bruzzone@unige.it [Department of Experimental Medicine, Section of Biochemistry, University of Genova, Viale Benedetto XV 1, 16132 Genova (Italy); Centre of Excellence for Biomedical Research, University of Genova, Viale Benedetto XV 9, 16132 Genova (Italy); Advanced Biotechnology Center, Largo Rosanna Benzi 10, 16132 Genova (Italy); Battaglia, Florinda [Centre of Excellence for Biomedical Research, University of Genova, Viale Benedetto XV 9, 16132 Genova (Italy); Mannino, Elena [Department of Experimental Medicine, Section of Biochemistry, University of Genova, Viale Benedetto XV 1, 16132 Genova (Italy); Parodi, Alessia [Centre of Excellence for Biomedical Research, University of Genova, Viale Benedetto XV 9, 16132 Genova (Italy); Fruscione, Floriana [Department of Experimental Medicine, Section of Biochemistry, University of Genova, Viale Benedetto XV 1, 16132 Genova (Italy); Advanced Biotechnology Center, Largo Rosanna Benzi 10, 16132 Genova (Italy); Basile, Giovanna [Department of Experimental Medicine, Section of Biochemistry, University of Genova, Viale Benedetto XV 1, 16132 Genova (Italy); Salis, Annalisa; Sturla, Laura [Department of Experimental Medicine, Section of Biochemistry, University of Genova, Viale Benedetto XV 1, 16132 Genova (Italy); Centre of Excellence for Biomedical Research, University of Genova, Viale Benedetto XV 9, 16132 Genova (Italy); Negrini, Simone; Kalli, Francesca; Stringara, Silvia [Centre of Excellence for Biomedical Research, University of Genova, Viale Benedetto XV 9, 16132 Genova (Italy); Filaci, Gilberto [Centre of Excellence for Biomedical Research, University of Genova, Viale Benedetto XV 9, 16132 Genova (Italy); Department of Internal Medicine, Viale Benedetto XV 6, 16132 Genova (Italy); and others

2012-05-25

342

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.

2012-05-25

343

Prolonged activation of ERK contributes to the photorejuvenation effect in photodynamic therapy in human dermal fibroblasts.  

Science.gov (United States)

Photodynamic therapy (PDT) is known to be effective in the photorejuvenation of photoaged skin. However, the molecular mechanisms of rejuvenation by PDT remain elusive. In this study, we aimed to understand the molecular events occurring during the photorejuvenation after PDT in dermal fibroblasts in vitro. First, we found that PDT conditions resulted in an increased fibroblast proliferation and motility in vitro. Under this condition, cells had increased intracellular reactive oxygen species (ROS) production. Importantly, PDT induced a prolonged activation of extracellular signal-regulated kinase (ERK) with a corresponding increase in matrix metalloproteinase (MMP)-3 and collagen type I? messenger RNA and protein. Moreover, inhibition of PDT-induced ERK activation significantly suppressed fibroblast proliferation and expression of MMP-3 and collagen type I? following PDT. In addition, NAC (an antioxidant) inhibited PDT-induced fibroblast proliferation and ERK activation indicating that prolonged ERK activation and intracellular ROS contribute to the proliferation of fibroblasts and the dermal remodeling process for skin rejuvenation. We also identified increased collagen volume and decreased elastotic materials that are used as markers of photoaging in human skin samples using histochemical studies. Results from this study suggest that intracellular ROS stimulated by PDT in dermal fibroblasts lead to prolonged activation of ERK and, eventually, fibroblast proliferation and activation. Our data thus reveal a molecular mechanism underlying the skin rejuvenation effect of PDT. PMID:23337889

Jang, Yong Hyun; Koo, Gi-Bang; Kim, Joo-Young; Kim, You-Sun; Kim, You Chan

2013-09-01

344

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, 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.)

2004-12-01

345

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 obtained indicate that extremely low-frequency electromagnetic fields has the potential for induction of chromosomal aberrations in all cell types.

2010-01-01

346

Analysis of the E-cadherin and P-cadherin promoters in murine keratinocyte cell lines from different stages of mouse skin carcinogenesis.  

Science.gov (United States)

We previously isolated the 5' upstream sequences of the mouse P-cadherin gene, in which putative binding sites for several transcription factors were identified between nt-101 and +30. In the study reported here, the promoter activity of the postulated 5' cis-acting sequences of the P-cadherin promoter, and the activity of the proximal E-cadherin promoter were investigated in several murine keratinocyte cell lines showing different levels of P- and E-cadherin expression as well as different morphology and tumorigenic behavior. Cell-type specificity and optimal activity of P-cadherin expression in murine keratinocytes was conferred by 5' sequences located between nt -200 and +30, and the GC-rich region (nt -101 to +80) and a CCAAT box element (nt -65) had a major regulatory role. The cell-type specificity of the E-cadherin promoter, on the other hand, was mediated by a combination of positive regulatory elements, a GC-rich region (nt -58 to -24), and a CCAAT box (nt -65) and repressor elements inside the E-pal sequence. Interestingly, the maximum repressor effect of the E-pal element was observed in non-expressing undifferentiated spindle cells. In vitro binding studies indicated that the GC-rich region of the P-cadherin promoter was mainly recognized by Sp1-related nuclear factors, whereas both AP2- and Sp1-related factors were involved in the interaction of the GC-rich region of the E-cadherin promoter. Common factors (probably related to the CP1 family) seemed also to be involved in the recognition of the CCAAT-box element of both the E- and P-cadherin promoters, but additional specific factors participated in the interaction with the CCAAT box of the E-cadherin promoter. Our studies also support the hypothesis that loss or modification of some of the regulatory factors occurs during mouse skin tumor progression. PMID:9328434

Faraldo, M L; Rodrigo, I; Behrens, J; Birchmeier, W; Cano, A

1997-09-01

347

Skin Graft  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

Shimizu, Ruka; Kishi, Kazuo

2012-01-01

348

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

349

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 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.)

2004-02-01

350

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 ...

351

Your Skin  

Science.gov (United States)

... BAY-shus) glands , and they are always producing sebum (say: SEE-bum). Sebum is your skin's own natural oil. It rises ... also makes your skin waterproof — as long as sebum's on the scene, your skin won't absorb ...

352

Immortalization of Werner syndrome and progeria fibroblasts  

International Nuclear Information System (INIS)

Human fibroblast cells from two different progeroid syndromes, Werner syndrome (WS) and progeria, were established as immortalized cell lines by transfection with plasmid DNA containing the SV40 early region. The lineage of each immortalized cell line was confirmed by VNTR analysis. Each of the immortalized cell lines maintained its original phenotype of slow growth. DNA repair ability of these cells was also studied by measuring sensitivity to killing by uv or the DNA-damaging drugs methyl methansulfonate, bleomycin, and cis-dichlorodiamine platinum. The results showed that both WS and progeria cells have normal sensitivity to these agents

1991-01-01

353

Immortalization of Werner syndrome and progeria fibroblasts  

Energy Technology Data Exchange (ETDEWEB)

Human fibroblast cells from two different progeroid syndromes, Werner syndrome (WS) and progeria, were established as immortalized cell lines by transfection with plasmid DNA containing the SV40 early region. The lineage of each immortalized cell line was confirmed by VNTR analysis. Each of the immortalized cell lines maintained its original phenotype of slow growth. DNA repair ability of these cells was also studied by measuring sensitivity to killing by uv or the DNA-damaging drugs methyl methansulfonate, bleomycin, and cis-dichlorodiamine platinum. The results showed that both WS and progeria cells have normal sensitivity to these agents.

Saito, H.; Moses, R.E. (Baylor College of Medicine, Houston, TX (USA))

1991-02-01

354

Comparison of Gene Expression Profiles between Keratinocytes, Melanocytes and Fibroblasts  

Science.gov (United States)

Background The skin has many important functions such as protection, preservation, temperature regulation, and vitamin D synthesis. It is composed of a variety of cell types including keratinocytes, melanocytes and fibroblasts. Objective We attempted to compare the gene expression profiles between keratinocytes, melanocytes and fibroblast, using cDNA microarray. Methods Keratinocytes, melanocytes and fibroblasts were primary cultured from five foreskin specimens. Total RNAs were extracted and pooled to reduce the individual variations, and then used for cDNA microarray. Results Total 12,028 genes were selected as the reliable genes whose expression was detected in at least one of the three cell types. By comparing the relative expression levels with cutoff limitation as a fourfold change, we obtained 126 fibroblast-specific, 179 keratinocyte-specific and 173 melanocyte-specific genes, many of which are known to be characteristically expressed in each cell type. In addition, we identified many genes whose skin-specific functions have not yet been determined. Conclusion Our data provide important information on which to base further investigation into the specification of skin cell types.

Lee, Jung-Suk; Kim, Dae-Hun; Choi, Dae-Kyoung; Kim, Chang Deok; Ahn, Gwang-Bum; Yoon, Tae Young; Lee, Jeung-Hoon

2013-01-01

355

Salmon and king crab trypsin stimulate interleukin-8 and matrix metalloproteinases via protease-activated receptor-2 in the skin keratinocytic HaCaT cell line.  

Science.gov (United States)

Occupational skin symptoms are prevalent among the workers of the seafood processing industry. In this study we investigate the role of salmon (Salmo salar) and king crab trypsin (Paralithodes camtschaticus) as inducers of inflammation in skin via secretion of inflammatory mediators. Human skin keratinocytes (HaCaT cells) were exposed to purified salmon and king crab trypsin. We observed that salmon trypsin enhanced the secretion of IL-8 and MMP-2 and crab trypsin enhanced the secretion of IL-8, MMP-2 and MMP-9 in a dose dependent manner. As protease activated receptors (PAR)-2 in skin are known to play an important role in physiology and pathology, we explored the involvement of these receptors in mediating the release of interleukin (IL)-8 and matrix metalloproteinase (MMP)-2 and -9 subsequent to exposure of skin keratinocytes to salmon and crab trypsin. In addition we observed that salmon and crab trypsin exhibit individual differences in stimulating the release of these inflammatory mediators. Finally, using specific small interfering RNA (siRNA) against PAR-2, we confirmed that the increase in secretion of IL-8, MMP-2 and MMP-9 in skin keratinocytes following exposure to salmon and crab trypsin was mediated via activation of PAR-2. These results suggest that exposure to proteases from the seafood may lead to inflammatory reactions in skin. PMID:24795235

Bhagwat, Sampada S; Larsen, Anett K; Winberg, Jan-Olof; Seternes, Ole-Morten; Bang, Berit E

2014-07-01

356

Contact sensitizer nickel sulfate activates the transcription factors NF-kB and AP-1 and increases the expression of nitric oxide synthase in a skin dendritic cell line  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Nuclear factor kappa B (NF-kB) and activating protein-1 (AP-1) transcription factors are ubiquitously expressed signaling molecules known to regulate the transcription of a large number of genes involved in immune responses, namely the inducible isoform of nitric oxide synthase (iNOS). In this study, we demonstrate that a fetal skin-derived dendritic cell line (FSDC) produces nitric oxide (NO) in response to the contact sensitizer nickel sulfate (NiSO4) and increases the ...

Cruz, M. Teresa; Gonc?alo, Margarida; Figueiredo, Ame?rico; Carvalho, Arse?lio P.; Duarte, Carlos B.; Lopes, M. Celeste

2004-01-01

357

Nerve growth factor levels in cultured human skin cells: effect of gestation and viral transformation.  

Science.gov (United States)

Extracts of cultured human keratinocytes and fibroblasts were assayed for nerve growth factor-like immunoreactivity (NGF) by a specific enzyme-linked immunoabsorbant assay. NGF levels were higher in primary cultured keratinocytes than in freshly isolated keratinocytes or culture through multiple passages. Viral transformation of keratinocytes with the human papilloma virus (HPV16) significantly increased NGF levels, whilst transformation with the simian virus (SV40), which induces simple epithelial differentiation, reduced the concentration of NGF. Passaged epidermal keratinocytes contained more than twice as much NGF as did passaged fibroblasts. Oral keratinocytes and fibroblasts, and psoriatic fibroblasts, all from high turnover tissues, did not contain significantly different levels of NGF in culture than dermal keratinocytes or fibroblasts. Foetal fibroblasts contained five times as much NGF as did adult fibroblasts. These results suggest that basal keratinocytes are a major but not sole source of NGF in human skin, and that NGF may play a role in human skin development. PMID:7715836

Anand, P; Foley, P; Navsaria, H A; Sinicropi, D; Williams-Chestnut, R E; Leigh, I M

1995-01-30

358

Skin decontamination  

International Nuclear Information System (INIS)

The problem matter is briefly characterized of skin damage by irradiation and of the binding of radioactive materials to the skin. The results are summed up of some experiments aimed at skin resorption of various radioactive materials. Main attention is devoted to methods of skin decontamination. The principles are described in detail of the operation of the so-called hygiene loops where complete decontamination of personnel proceeds. The organizational chart of such a unit is presented in figure. Information is also presented on the composition of solutions for skin decontamination. (Z.M.)

1986-08-20

359

Age-related skin changes.  

Science.gov (United States)

Age-related skin changes can be induced by chronological ageing, manifested in subcutaneous fat reduction, and photo-ageing eliciting increased elastotic substance in the upper dermis, destruction of its fibrilar structure, augmented intercellular substance and moderate inflammatory infiltrate. Forty-five biopsy skin samples of the sun-exposed and sun-protected skin were analyzed. The patients were both males and females, aged from 17 to 81 years. The thickness of the epidermal layers and the number of cellular living layers is greater in younger skin. The amount of keratohyaline granules is enlarged in older skin. Dermoepidermal junction is flattened and the presence of elastotic material in the dermis is pronounced with age. The amount of inflammatory infiltrate is increased, the fibrous trabeculae are thickened in older skin and the atrophy of the hypodermis is observed. Chronological ageing alters the fibroblasts metabolism by reducing their life span, capacity to divide and produce collagen. During ageing, the enlargement of collagen fibrils diminishes the skin elasticity. PMID:22730701

Levakov, Aleksandra; Vuckovi?, Nada; Dolai, Matilda; Ka?anski, Mihaela Mocko; Bozani?, Snezana

2012-01-01

360

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

 
 
 
 
361

Mechanical Tension Increases CCN2/CTGF Expression and Proliferation in Gingival Fibroblasts via a TGF?-Dependent Mechanism  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Unlike skin, oral gingival do not scar in response to tissue injury. Fibroblasts, the cell type responsible for connective tissue repair and scarring, are exposed to mechanical tension during normal and pathological conditions including wound healing and fibrogenesis. Understanding how human gingival fibroblasts respond to mechanical tension is likely to yield valuable insights not only into ...

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

2011-01-01

362

HSP27 regulates fibroblast adhesion, motility, and matrix contraction  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Heat shock protein 27 (HSP27) modulates actin-dependent cell functions in several systems. We hypothesized that HSP27 modulates wound contraction. Stably transfected fibroblast cell lines that overexpress HSP27 (SS12) or underexpress HSP27 (AS10) were established, and cell behaviors related to wound contraction were examined. First, fibroblast-populated collagen lattice (FPCL) contraction was examined because it has been studied as a wound-healing model. In floating FPCL contraction assays, S...

Hirano, Sahoko; Shelden, Eric A.; Gilmont, Robert R.

2004-01-01

363

Purification of the migration stimulating factor produced by fetal and breast cancer patient fibroblasts  

International Nuclear Information System (INIS)

The authors have previously shown that (i) human skin fibroblasts of fetal and adult origin display distinctive migratory phenotypes, (ii) this difference in cell behavior results from the production of a soluble migration stimulating factor (MSF) by fetal cells, and (iii) skin fibroblasts from breast cancer patients commonly resemble fetal fibroblasts both in migratory phenotype and in production of MSF. Data are now presented indicating that MSF present in the conditioned medium of fetal and cancer patient fibroblasts is precipitated at 10% saturation ammonium sulfate and binds to heparin and cation-exchange resins. Based on this information, they have devised a scheme for the purification of MSF involving the sequential application of ammonium sulfate precipitation, heparin affinity, gel filtration, and reverse-phase chromatography. Purified MSF has an estimated molecular mass of 70 kDa; amino acid analysis reveals a relatively high level of proline (13.34 residues per 100). The results further suggest that skin fibroblasts from breast cancer patients produce an additional factor with migration stimulating activity; this factor is precipitated at higher concentrations of ammonium sulfate and binds to anion-exchange resins. They have previously discussed the possible direct involvement of fetal-like fibroblasts in cancer pathogenesis. The availability of MSF obtained from cancer patient fibroblasts provides a potential means with which to examine the complex cellular interactions contributing to this process as well as develop a screening regime for identifying individuals at elevated risk of developing cancer

1989-01-01

364

Effect of estrogens on skin aging and the potential role of SERMs  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In humans, structural and functional changes attributable to aging are more visibly evident in the skin than in any other organ. Estrogens have significant effects on skin physiology and modulate epidermal keratinocytes, dermal fibroblasts and melanocytes, in addition to skin appendages including the hair follicle and the sebaceous gland. Importantly, skin aging can be significantly delayed by the administration of estrogen. This paper reviews the effects of estrogens on skin and the mechanis...

2007-01-01

365

The nano-scale mechanical properties of the extracellular matrix regulate dermal fibroblast function.  

Science.gov (United States)

Changes in the mechanical properties of dermis occur during skin aging or tissue remodeling and affect the activity of resident fibroblasts. With the aim to establish elastic culture substrates that reproduce the variable softness of dermis, we determined Young's elastic modulus E of human dermis at the cell perception level using atomic force microscopy. The E of dermis ranged from 0.1 to 10?kPa, varied depending on body area and dermal layer, and tended to increase with age in 26-55-year-old donors. The activation state of human dermal fibroblasts cultured on "skin-soft" E (5?kPa) silicone culture substrates was compared with stiff plastic culture (GPa), collagen gel cultures (0.1-9?kPa), and fresh human dermal tissue. Fibroblasts cultured on skin-soft silicones displayed low mRNA levels of fibrosis-associated genes and increased expression of the matrix metalloproteinases (MMPs) MMP-1 and MMP-3 as compared with collagen gel and plastic cultures. The activation profile exhibited by fibroblasts on "skin-soft" silicone culture substrates was most comparable with that of human dermis than any other tested culture condition. Hence, providing biomimetic mechanical conditions generates fibroblasts that are more suitable to investigate physiologically relevant cell processes than fibroblasts spontaneously activated by stiff conventional culture surfaces. PMID:24670384

Achterberg, Volker F; Buscemi, Lara; Diekmann, Heike; Smith-Clerc, Josiane; Schwengler, Helge; Meister, Jean-Jacques; Wenck, Horst; Gallinat, Stefan; Hinz, Boris

2014-07-01

366

Roscovitine inhibits differentiation and invasion in a three-dimensional skin reconstruction model of metastatic melanoma.  

Science.gov (United States)

The aim of this study was to investigate the therapeutic potential of a cyclin-dependent kinase inhibitor, roscovitine, in cultured melanoma cells and a three-dimensional skin reconstruction model of metastatic melanoma. The modulatory effects of roscovitine on the growth and survival of normal melanocytes and cultured melanoma cell lines were tested. Additionally, we investigated the potential of roscovitine to regulate the growth and differentiation of a metastatic melanoma cell line (A375) in a three-dimensional skin reconstruction culture consisting of A375 cells admixed with normal human keratinocytes embedded within a collagen-constricted fibroblast matrix. We show that roscovitine is able to induce apoptosis in the melanoma cell lines A375, 888, and 624 but not in normal human cultured epithelial melanocytes. The degree of apoptosis within these cell lines correlated with the accumulation of p53 protein and concomitant reduction of X-linked inhibitor of apoptosis protein, with no change in the proteins Bcl-2 and survivin. We also found that roscovitine inhibited the growth and differentiation of A375 melanoma cells within the dermal layer of the skin. The results of this study show that roscovitine has the potential to inhibit the differentiation and invasion of metastatic melanoma and may be useful as a therapy for the treatment of patients with metastatic melanoma. PMID:17314272

Mohapatra, Subhra; Coppola, Domenico; Riker, Adam I; Pledger, W Jack

2007-02-01

367

Ultrastructural features of composite skin cultures grafted onto athymic mice.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Skin substitutes composed of cultured keratinocytes with or without a dermal substrate are now being used in the treatment of burns and other cutaneous wounds. Composite skin cultures (Graftskin, LSE), consisting of epidermal keratinocytes seeded on a fibroblast-containing collagen matrix and maintained at the air-liquid interface, develop a well differentiated epidermis in vitro with many of the morphological and biochemical features of intact skin. Basement membrane-associated antigens, dev...

Nolte, C. J.; Oleson, M. A.; Hansbrough, J. F.; Morgan, J.; Greenleaf, G.; Wilkins, L.

1994-01-01

368

Human skin equivalent as an alternative to animal testing  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The 3-D skin equivalent can be viewed as physiologically comparable to the natural skin and therefore is a suitable alternative for animal testing. This highly differentiated in vitro human skin equivalent is used to assess the efficacy and mode of action of novel agents. This model is generated from primary human keratinocytes on a collagen substrate containing human dermal fibroblasts. It is grown at the air-liquid interface which allows full epidermal stratification and epidermal-dermal in...

2008-01-01

369

Decreased host cell reactivation of UV-irradiated adenovirus 5 by fibroblasts from Cockayne's syndrome patients  

International Nuclear Information System (INIS)

Over a period of 5 years, 29 experiments were performed in which survival curves of UV-irradiated adenovirus were determined using fibroblast strains from 10 normal persons and from 7 persons having Cockayne's syndrome. In all of these, the survival of UV-irradiated adenovirus 5 was less when assayed using monolayers of fibroblasts from Cockayne's syndrome patients than from normal persons. Survival curves using normal fibroblasts were, within error, straight lines on a log survival vs. linear fluence plot. Survival curves obtained using Cockayne's syndrome fibroblasts showed 2 components: an initial sensitive component, reflecting the behavior of approx. 75% of the infected cells, followed by a component having normal sensitivity. In the 28 experiments that were considered reliable, 58 curves were done using Cockayne's fibroblasts, 41 using normal human fibroblasts. Although experimental variation was encountered, there was no individual case in which sensitivity as measured using Cockayne's was equal to (or less than) the sensitivity measured using normal fibroblasts. (author)

1981-01-01

370

Curious Skin  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Some of Henry Wellcome’s collection of tattoos on human skin will be on display in our forthcoming Skin exhibition. But how did the Parisian doctor from whom they were acquired come by his macabre collection of tattoos in the first place, and what did they mean to those whose skin they were on? It’s Gemma Angel‘s job to find out…

Angel, G.

2010-01-01

371

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

372

Dynamic Tensile Properties of Human Skin  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

2012-01-01

373

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.)

1978-03-17

374

Modulation of cellular senescence in fibroblasts and dermal papillae cells in vitro.  

Science.gov (United States)

A hexapeptide (Hexapeptide-11) of structure Phe-Val-Ala-Pro-Phe-Pro (FVAPFP) originally isolated from yeast extracts and later synthesized by solid state synthesis to high purity has demonstrated an ability to influence the onset of senescence in intrinsically aged fibroblasts, extrinsically aged fibroblasts, and extrinsically aged dermal papillae cells in vitro. The mechanism of senescence control is believed to be related to the peptide's ability to reversibly downregulate ataxia telangiectasia mutated (ATM) and p53 protein expression. The importance of p53 as the gatekeeping protein for monitoring cellular DNA damage is strategic for maintaining cellular health. ATM activates p53 by direct phosphorylation, causing cells to move into senescence which effectively moves them out of reproductive processes. Technologies that can influence ATM and p53 expression may offer unique benefits for controlling cellular senescence and effectively delaying cellular aging processes. The influence on ATM and p53 expression is noted to occur in both cell lines at peptide concentrations between 0.1% and 1.0%. The implications of these effects for aging benefits for skin and hair is important as, to date, no known small peptide has been suggested to demonstrate this effect in such a reversible and dose-dependent fashion. PMID:23578831

Gruber, James V; Ludwig, Philip; Holtz, Robert

2013-01-01

375

Repair of DNA strand breaks in progeric fibroblasts and aging human diploid cells  

International Nuclear Information System (INIS)

The rate of rejoining of DNA strand breaks induced by 10 krad of ?-irradiation has been studied in normal human diploid skin fibroblasts and skin fibroblasts from six patients with symptoms of progeria. Although slightly more rapid in very early passage, the repair rate in normal cells was similar throughout most of their life span in vitro. The appearance of cells with reduced repair capacity was evident as the cultures became senescent. The progeric fibroblasts varied greatly in their response to irradiation. The rate of repair was greatly reduced in two strains, whereas in two others extensive DNA degradation was consistently observed in unirradiated cells. Degradation was apparently related to the radiation received from the incorporated radiolabel. Normal repair was seen in progeric fibroblasts transformed by SV40 virus

1975-01-01

376

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 fo