Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Human dermal fibroblasts”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 127 records · Page 7Linked to original sources
PubMed1993

Interleukin-8 production from cultured human dermal fibroblasts by stimulation with supernatant of cultured human epidermal cells.

There is much evidence to support the theory that keratinocytes and dermal fibroblasts actively participate in inflammatory reactions by the production of proinflammatory mediators or cytokines. We investigated the neutrophil chemotactic activity in conditioned media of cultured epidermal cells and dermal fibroblasts, and found that an epidermal cell-derived factor induced fibroblasts to produce a neutrophil chemotactic factor. This neutrophil chemotactic factor was identified as interleukin-8 (IL-8) by the elution position on HPLC and by a neutralization test that uses monoclonal anti-IL-8 antibody (14E4). The epidermal cell-derived factor was fractionated together with thymocyte-proliferating activity on Sephadex G-75 gel chromatography followed by HPLC. It was blocked specifically by anti-interleukin-1 (IL-1) alpha antibody, which indicates that this factor was IL-1 alpha. Since a variety of inflammatory dermatoses is characterized by the infiltration of neutrophils into the skin, induction of IL-8 production in fibroblasts by epidermal cells may play an important role in inflammatory skin diseases.

Cells, Cultured↗
PubMed1992

Culture of human dermal fibroblasts in collagen gels: modulation of interleukin 1-induced prostaglandin E2 synthesis by an extracellular matrix.

Human dermal fibroblasts, cultured as suspensions in collagen gels and as monolayers, were stimulated with recombinant human interleukin-1 beta (rIL 1 beta) at 72 h, and prostaglandin E2 (PGE2) was assayed 24 h later. Fibroblasts in gels were less responsive to rIL 1 beta than monolayers, PGE2 synthesis increasing from less than 1 ng/microgram DNA without rIL 1 beta to maxima of 11.3 and 32.9 ng/micrograms DNA, with half maximal release occurring at 7.47 and 0.75 pM rIL 1 beta for the gel and monolayer cultures, respectively. Increased PGE2 was first detected 4 h after addition of rIL 1 beta to gels and was inhibited by 10(-5) M indomethacin. The amount of PGE2 synthesized per fibroblast increased with the time the gels had been in culture when stimulated with rIL 1 beta and was proportional to the number of fibroblasts in the gels, but inversely related to the collagen concentration. A common feature of these experiments was significantly greater induction of PGE2 synthesis at higher cell densities in collagen gels. Exogenous 10(-4) M arachidonic acid further increased PGE2 synthesis by rIL 1 beta-stimulated fibroblasts, but the differential in the amount of PGE2 released from fibroblasts at high and low population densities in the gels was maintained. These results are consistent with interleukin 1 (IL 1) stimulating PGE2 synthesis in dermal fibroblasts by increasing cyclooxygenase activity. Furthermore, the results show that dermal fibroblasts have an additional regulatory mechanism, related to the cell population densities or their interactions with an extracellular matrix, to finely modulate the amount of PGE2 synthesized in response to IL 1.

Arachidonic Acid↗
PubMed1988

Tumor necrosis factor inhibits collagen and fibronectin synthesis in human dermal fibroblasts.

Tumor necrosis factor (TNF) caused inhibition of collagen production by confluent cultures of human dermal fibroblasts in a dose-dependent manner. Concomitant increase of prostaglandin E2 release was observed as a result of TNF-induced cell activation. However, a blockade of the cyclooxygenase pathway of arachidonate metabolism by indomethacin did not abrogate the inhibitory effect of TNF on collagen synthesis, suggesting that this effect could be independent of prostaglandin metabolism. Gel electrophoresis of the newly synthesized macromolecules from the culture media showed that both type I and type III collagens as well as fibronectin were affected by the inhibition. Electrophoresis of cell layer-associated proteins demonstrated that the reduction in amounts of collagen and fibronectin in the medium did not result from an intracellular accumulation of these macromolecules. Production of procollagens was reduced in parallel to that of collagens, suggesting that the effect of TNF is exerted before the processing steps of procollagens. These results clearly show that TNF could play a role in modulation of matrix deposition by fibroblasts during inflammatory processes.

Cell Division↗
PubMed2005

Stimulation of mitochondrial activity by p43 overexpression induces human dermal fibroblast transformation.

Mitochondrial dysfunctions are frequently reported in cancer cells, but their direct involvement in tumorigenesis remains unclear. To understand this relation, we stimulated mitochondrial activity by overexpression of the mitochondrial triiodothyronine receptor (p43) in human dermal fibroblasts. In all clones, this stimulation induced morphologic changes and cell fusion in myotube-like structures associated with the expression of several muscle-specific genes (Myf5, desmin, connectin, myosin, AchRalpha). In addition, these clones displayed all the in vivo and in vitro features of cell transformation. This phenotype was related to an increase in c-Jun and c-Fos expression and extinction of tumor suppressor gene expression (p53, p21WAF1, Rb3). Lastly, reactive oxygen species (ROS) production was increased in positive correlation to the stimulation of mitochondrial activity. The direct involvement of mitochondrial activity in this cell behavior was studied by adding chloramphenicol, an inhibitor of mitochondrial protein synthesis, to the culture medium. This inhibition resulted in partial restoration of the normal phenotype, with the loss of the ability to fuse, a strong decrease in muscle-specific gene expression, and potent inhibition of the transformed phenotype. However, expression of tumor suppressor genes was not restored. Similar results were obtained by using N-acetylcysteine, an inhibitor of ROS production. These data indicate that stimulation of mitochondrial activity in human dermal fibroblasts induces cell transformation through events involving ROS production.

Animals↗
PubMed2003

Human dermal fibroblasts do not exhibit directional migration on collagen I in direct-current electric fields of physiological strength.

Endogenous electric fields are generated lateral to skin wounds, with the cathodal pole of the field residing in the center of the wound. These fields are thought to be an important mechanism in guiding the migration of keratinocytes and other cells into wounds to effect healing. In this work, human dermal fibroblasts were exposed to direct current electric fields of physiological strength, and their migrational behavior was quantitated. Only random migration of human dermal fibroblasts was observed in direct-current electric fields under conditions that support the directional migration of human epidermal keratinocytes. Additionally, neither the presence of serum nor serum plus additional Mg++ in the experimental medium supported directional migration. Migratory rates of fibroblasts varied depending on the experimental medium used: in serum-containing medium the average velocity was as low as 0.23 micro m/min, while in serum-free keratinocyte medium the average velocity was as high as 0.36 micro m/min. These studies suggest that dermal fibroblasts do not respond to the endogenous electric field of a wound, and use other migratory cues to direct their movement into the wound bed.

Cell Movement↗
PubMed1998

Regulation of RANTES and IL-8 production in normal human dermal fibroblasts by active vitamin D3 (tacalcitol).

1. The production of chemokines, RANTES and IL-8 in cultured human dermal fibroblasts and the effects of tacalcitol (1alpha,24(R)-dihydroxyvitamin D3) were studied using an enzyme-linked immunosorbent assay. 2. In the unstimulated condition, RANTES and IL-8 were at a trace level in the culture supernatant. On stimulation with TNF-alpha alone for 24 h, RANTES and IL-8 production were induced. Tacalcitol suppressed RANTES and IL-8 production dose-dependently at concentrations between 10(-12) M and 10(-7) M. 3. When the cells were treated with TNF-alpha and IFN-gamma in combination, RANTES production was enhanced, but IL-8 production was not changed, compared to TNF-alpha-treated cells. Tacalcitol decreased IL-8 production dose-dependently as observed in the TNF-alpha-treated cells. On the other hand, RANTES production was enhanced by 10(-11) M and 10(-10) M of tacalcitol, and dose-dependently suppressed by tacalcitol concentrations higher than 10(-9) M. 4. Active vitamin D3 compounds, betamethasone valerate and cyclosporin A were compared with respect to their effects on chemokine production. Three active vitamin D3 compounds, tacalcitol, 1alpha,25-dihydroxyvitamin D3 and MC903 (calcipotriol), inhibited the production of RANTES and IL-8, with very similar potencies. Betamethasone valerate also inhibited these chemokine productions, but with greater potency than active vitamin D3 compounds. Cyclosporin A significantly stimulated RANTES production at 10(-6) M and IL-8 production at 10(-7) M and 10(-6) M. 5. The results of this study suggest that active vitamin D3 compounds exert some beneficial effects in the treatment of inflammatory skin diseases via regulation of the production of chemokines by dermal fibroblasts.

Betamethasone↗
PubMed1998

Modulation of basal expression of the human alpha1(I) procollagen gene (COL1A1) by tandem NF-1/Sp1 promoter elements in normal human dermal fibroblasts.

We previously demonstrated that a segment of the human alpha1 type I procollagen gene (COL1A1) promoter encompassing nt -174 to -84 is responsible for the highest transcriptional activity in collagen producing cells in vitro. Here, we identified two almost identical tandem NF-1/Sp1 binding sites located between nt -129 to -107 (distal element) and nt -104 to -77 (proximal element) that are responsible for the basal regulation of COL1A1 transcription in normal human dermal fibroblasts. Transient transfection studies revealed that 85% of the basal COL1A1 promoter activity resides within the distal element; however, site-directed mutagenesis within the CCAAT motif in the proximal element resulted in a 98% decrease of the COL1A1 promoter activity. We conclude that each of the NF-1/Sp1 tandem binding sites has a different function. The distal element drives the transcriptional activity of the COL1A1 promoter but is not sufficient for its basal expression, whereas the NF-1 binding site in the proximal element is essential for in vitro COL1A1 gene transcription.

Base Sequence↗
PubMed1996

Juxtacrine stimulation of cytokine production in cocultures of human dermal fibroblasts and T cells.

Adhesion of T cells to fibroblasts activates cells to produce cytokines, either by direct cell contact and/or soluble factors. A cell-associated form of IL-1 beta on fibroblasts might act through a cell contact mediated fashion. To test this hypothesis we analysed the activation of T cells and human dermal fibroblasts (HDF) in coculture experiments. Elevated levels of IL-1 beta, secreted by T cells as well as IL-6 and IL-8, mainly produced by HDF, were found in supernatant fluids of cocultured cells. IL-1 beta mRNA expression was induced in T cells as well as in HDF. While in HDF IL-1 beta remained cell-associated, T cells were activated to produce and secrete soluble IL-1 beta and IL-6. IL-1 beta and possibly other soluble factors increased IL-6 production by fibroblasts. These effects could be mainly attributed to CD8+ T cells. Our results suggest, that IL-1 beta, produced as a cell-associated cytokine by human dermal fibroblasts, acts as a juxtacrine molecule to stimulate T cells. Such a cellular cooperation, could be a powerful mediator in inflammatory response and possibly in wound healing.

Coculture Techniques↗
PubMed2000

Effect of mast cell-derived mediators and mast cell-related neutral proteases on human dermal fibroblast proliferation and type I collagen production.

BACKGROUND: Possible involvement of mast cells in various fibrotic conditions has been suggested, but the relative contribution of each mast cell mediator and neutral protease to fibroproliferative activity remains to be elucidated. OBJECTIVE: We investigated the effect of mast cell-derived mediators and mast cell-related neutral proteases on type I collagen production and proliferation by human dermal fibroblasts. METHODS: Mast cell-derived mediators or neutral proteases were added to cultured fibroblasts from normal dermis, and cell proliferation and type I collagen synthesis were assayed. RESULTS: Fibroblast proliferation was increased by 2.8 x 10(-9) mol/L prostaglandin D(2) and 10 microgram/mL carboxypeptidase A, but not by leukotriene D(4) or cathepsin G at the concentrations studied. Proliferation was increased by tryptase in a concentration-dependent manner, and a significant increase was observed at concentrations of 1 and 10 microgram/mL. Production of type I collagen by fibroblasts was increased in the presence of 2.0 x 10(-9) mol/L leukotriene D(4) and 10 microgram/mL tryptase. CONCLUSION: Mast cell-derived mediators prostaglandin D(2) and leukotriene D(4) and mast cell-related neutral proteases carboxypeptidase A and tryptase increase the proliferation and type I collagen production of human dermal fibroblasts in various manners.

Cell Division↗
PubMed1996

Chondroinduction of human dermal fibroblasts by demineralized bone in three-dimensional culture.

We designed and tested a three-dimensional device composed of demineralized bone powder (DBP) packed within a bilaminate sponge as an in vitro model for chondroinduction induced by DBP. Control bovine articular chondrocytes deposited cartilage matrix when cultured in these sponges. Human dermal fibroblasts that were seeded onto the composite sponge migrated through the collagen lattice and attached to the particles of DBP. Fibroblasts surrounding the DBP deposited a fine, granular extracellular matrix characterized by metachromatic staining similar to that deposited around the bona fide chondrocytes. Immunohistological analysis showed that the matrix contained chondroitin delta Di-4 sulfate, chondroitin delta Di-6 sulfate, and keratan sulfate epitopes. These components accumulated during the culture period, as quantified by immunochemical analysis of extracted matrix. In contrast, fibroblasts cultured in collagen sponges without DBP did not produce such a cartilage matrix. Control bovine chondrocytes produced these matrix molecules when cultured in the presence or absence of DBP. Thus this composite three-dimensional device facilitated evaluation of chondroinduction in human dermal fibroblasts in vitro.

Animals↗
PubMed1987

PGE2 causes a coordinate decrease in the steady state levels of fibronectin and types I and III procollagen mRNAs in normal human dermal fibroblasts.

Prostaglandin E2 (PGE2) caused inhibition of collagen and fibronectin synthesis by confluent cultures of human dermal fibroblasts. Dot-blot hybridization to cDNA probes complementary to Types I and III procollagens and fibronectin demonstrated that inhibition of protein production was accompanied by a coordinate decrease in the steady-state levels of the corresponding mRNAs. Blockade of transcription by actinomycin D demonstrated that PGE2 did not alter the stability of these mRNA. These results indicate that PGE2 is capable of exerting modulation of extracellular matrix biosynthesis, and that these effects occur at a transcriptional level.

Cells, Cultured↗
PubMed1998

Study of cytotoxicity mechanisms of silver nitrate in human dermal fibroblasts.

Certain concentrations of the antiseptic AgNO3, a potent broad-spectrum antimicrobial agent, exert cytotoxic effects on fibroblasts and endothelial cells which are directly related to the wound-healing process. In vitro assessment of human fibroblast cytotoxicity has proved to be a useful method for characterizing cell toxicity mechanisms of topically-applied antiseptics. In the present study human dermal fibroblasts were exposed to AgNO3 at concentrations of 4.12-82.4 microM for 8 and 24 h. Silver ions greatly inhibited fibroblast proliferation and prolonged AgNO3 exposure produced Ag-dependent cell loss. In the sequence of events occurring during our in vitro experimental model, the inhibitory action on DNA synthesis was the primary event in AgNO3 cytotoxicity, associated with significant loss of cell protein. Both parameters decreased as the content of fetal calf serum (FCS) in the exposure medium was gradually reduced from 10 to 2%. The parallel study of DNA synthesis and cell protein content suggests that the toxic damage produced by silver in different phases of the cell cycle may lead to destruction of the entire cell population and therefore hinder the tissue regeneration process. A concentration- and time-dependent depletion of intracellular ATP content is also caused by ionic silver, thereby compromising the cell energy charge which precedes human dermal fibroblast death. Concomitant incorporation of FCS to the medium always attenuated Ag+ cytotoxicity curves in a concentration-dependent fashion and maximum protection was observed at 10% in situations closely resembling physiological conditions.

Adenosine Triphosphate↗
PubMedDate not supplied

Nitric oxide promotes mitogen-induced dna synthesis in human dermal fibroblasts through cGMP.

1. Nitric oxide (NO) is a free radical with multiple functions in cellular pathophysiology. Nitric oxide has been proven to play an important role in wound healing; however, the mechanisms by which NO may promote wound healing are not clearly understood. We have investigated the effect of NO on growth factor-induced DNA synthesis in human dermal fibroblasts to suggest interactions between growth factors and NO as a possible mechanism for the role of NO in wound healing. 2. The NO donor sodium nitroprusside (SNP) significantly (P < 0.001) increased fetal bovine serum-induced thymidine incorporation into the DNA of human dermal fibroblasts. The maximal comitogenic concentration of SNP (100 micro mol/L) was also found to significantly (twofold; P < 0.01) enhance fibroblast growth factor- or platelet-derived growth factor-induced DNA synthesis. 3. Nitric oxide treatment significantly increased the production of cGMP. 8-Bromo-cGMP, a stable structural analogue of cGMP, was found to markedly potentiate (P < 0.001) the growth factor-induced DNA synthesis. 4. This study concludes that NO and cGMP promote growth factor-induced DNA synthesis in dermal fibroblasts, suggesting another possible mechanism by which NO may promote skin wound healing.

Cells, Cultured↗
PubMed2002

Increase in gap-junctional intercellular communications (GJIC) of normal human dermal fibroblasts (NHDF) on surfaces coated with high-molecular-weight hyaluronic acid (HMW HA).

Normal human dermal fibroblast (NHDF) cells were used to detect differences in gap-junctional intercellular communication (GJIC) by hyaluronic acid (HA), a linear polymer built from repeating disaccharide units that consist of N-acetyl-D-glucosamine (GlcNa) and D-glucuronic acid (GlcA) linked by a beta 1-4 glycosidic bond. The NHDF cells were cultured with different molecular weights (MW) of HA for 4 days. The rates of cell attachment in dishes coated with high-molecular-weight (HMW; 310 kDa or 800 kDa) HA at 2 mg/dish were significantly reduced at an early time point compared with low-molecular-weight (LMW; 4.8 kDa or 48 kDa) HA with the same coating amounts. HA-coated surfaces were observed by atomic force microscopy (AFM) under air and showed that HA molecules ran parallel in the dish coated with LMW HA and had an aggregated island structure in the dish coated with HMW HA surfaces. The cell functions of GJIC were assayed by a scrape-loading dye transfer (SLDT) method using a dye solution of Lucifer yellow. Promotion of the dye transfer was clearly obtained in the cell monolayer grown on the surface coated with HMW HA. These results suggest that HMW HA promotes the function of GJIC in NHDF cells. In contrast, when HMW HA was added to the monolayer of NHDF cells, the functions of GJIC clearly were lowered in comparison with the cells grown in the control dish or with those grown on the surface of HMW HA. Therefore it is concluded that the MW size of HA and its application method are important factors for generating biocompatible tissue-engineered products because of the manner in which the GJIC participates in cell differentiation and cell growth rate.

Cell Adhesion↗
PubMed2006

Fibronectin functional domains coupled to hyaluronan stimulate adult human dermal fibroblast responses critical for wound healing.

Fibronectin (FN) facilitates dermal fibroblast migration during normal wound healing. Proteolytic degradation of FN in chronic wounds hampers healing. Previously, three FN functional domains (FNfd) have been shown to be sufficient for optimal adult human dermal fibroblast migration. Here we report the development of an acellular hydrogel matrix comprised of the FNfds coupled to a hyaluronan (HA) backbone to stimulate wound repair. Employing Michael-type addition, the cysteine- tagged FNfds were first coupled to a homobifunctional PEG derivative. Thereafter, these PEG derivative FNfd solutions, containing bifunctional PEG-derivative crosslinker were coupled to thiol-modified HA (HA-DTPH) to obtain a crosslinked hydrogel matrix. When evaluated in vitro, these acellular hydrogels were completely cytocompatible. While spreading and proliferation of adult human dermal fibroblasts plateaued at higher FNfd bulk densities, their rapid and robust migration followed a typical bell-shaped response. When implanted in porcine cutaneous wounds, these acellular matrices, besides being completely biocompatible, induced rapid and en masse recruitment of stromal fibroblasts that was not observed with RGD-tethered or unmodified hydrogels. Such constructs might be of great benefit in clinical settings where rapid formation of new tissue is needed.

Adult↗
PubMed2003

Growth factor profile of irradiated human dermal fibroblasts using a serum-free method.

Radiation therapy for cancer permanently damages tissue in the line of treatment. This study sought to establish a serum-free protocol to evaluate the growth of irradiated fibroblasts and to analyze the levels of basic fibroblast growth factor (bFGF) and transforming growth factor-beta (TGF-beta) compared with normal fibroblasts. One irradiated cell line of human dermal fibroblasts was established from an intraoperative specimen obtained from a patient who had undergone radiation therapy for head and neck cancer. Irradiated and normal fibroblasts were then plated in UltraCULTURE (serum and growth factor free), modified Webber's medium (bFGF 50 ng/ml, insulin-like growth factor 100 ng/ml), and Dulbecco's Modified Eagle Medium with 10% fetal bovine serum (serum with undefined basal growth factors). Irradiated cells were also seeded in UltraCULTURE with 50 and 100 ng/ml of bFGF. Cell counts were performed at 0, 1, 3, 5, and 7 days, and cell supernatants were assayed for bFGF and TGF-beta. Irradiated and normal fibroblasts exhibited stronger growth in modified Webber's medium than in Dulbecco's Modified Eagle Medium with 10% fetal bovine serum. Growth of irradiated fibroblasts under bFGF modulation was similar to their growth in Webber's medium. Furthermore, irradiated fibroblasts remained viable in a serum-free and growth factor-free environment for at least 7 days; however, their growth and autocrine growth factor production was less than that of normal cells. This confirms the results of previous studies suggesting that cells from irradiated tissue undergo cellular changes. This study provides an effective model for the first-line evaluation of agents to improve wound healing, and it helps to establish standard levels of bFGF and TGF-beta production for irradiated fibroblasts.

Cell Count↗
PubMed2004

Triterpenoid from Tiarella polyphylla, regulation of type 1 procollagen and MMP-1 in ultraviolet irradiation of cultured old age human dermal fibroblasts.

Although many studies have been performed to elucidate the molecular consequences of ultraviolet irradiation, little is known about the effect of natural products. Ultraviolet irradiation is widely considered to be an environmental stress. Here we investigated the effect of 3,23-dihydroxy-20(29)-lupen-27-oic acid on the regulation of MMP-1 and type 1 procollagen in Ultraviolet irradiation of cultured old age human dermal fibroblasts. 3, 23-dihydroxy-20(29)-lupen-27-oic acid was isolated from Tiarella polyphylla D. Don (Saxifragaceae). Among them, 3, 23-dihydroxy-20(29)-lupen-27-oic acid induced the regulation of Type 1- procollagen and reduced the regulation of MMP-1 at the protein levels in a dose-dependent manner by ultraviolet irradiation. Taken together, our results suggest that 3, 23-dihydroxy-20(29)-lupen-27-oic acid plays an important role in the induction of Type 1-procollagen and reduction of MMP-1 by ultraviolet irradiation in old age human dermal fibroblasts.

Aged↗
PubMed2005

Alpha-melanocyte stimulating hormone cytoprotective biology in human dermal fibroblast cells.

Alpha-melanocyte stimulating hormone (alpha-MSH) has been identified as a potent anti-inflammatory peptide effective in various tissues including skin. It acts by inhibiting the production and action of several pro-inflammatory stimuli including TNF-alpha, IL-1beta and LPS in a number of cell types. The role of such stimuli in inducing cellular apoptosis is also well described; however the precise role of alpha-MSH in apoptosis is presently unclear, with studies reporting both anti- and pro-apoptotic activity. The present study demonstrates that cultured human dermal fibroblasts respond to serum depletion and TNF-alpha, IL-1beta and LPS with an increase in membrane permeability, a decrease in viability and an increase in phosphatidylserine externalization (indicative of apoptosis) over 48-96 h. alpha-MSH (at 10(-6) M, but not 10(-9) M) was found to inhibit the serum free and pro-inflammatory mediated reduction in membrane permeability and cellular viability and also inhibited increases in apoptosis. In conclusion, data support a cytoprotective and anti-apoptotic role of the alpha-MSH peptide in human dermal fibroblast cells.

Apoptosis↗