Papers by Keyword: Dermal Fibroblasts

Abstract: The aim of this research is using different dose of UVB radiation to study the influence on dermal fibroblasts. These studies include cell viability analysis, proliferation rate and Youngs modulus measurement using Atomic force microscope (AFM). Results have exhibited that the cell viability was 100%, 89.46%, 73.76% and 70.75% when the exposed dose of UVB radiation on dermal fibroblasts was 0, 540, 1080 and 2160mJ/cm². After 24 hours of incubation, dermal fibroblasts without exposure to UVB radiation increased cell growth to 183.5% but cells exposed to UVB radiation between 216 and 2160 mJ/cm² reduced cell growth rate to 131% and 47.7%. Moreover, the Youngs modulus of dermal fibroblasts measured by AFM ranged from 24.12 to 9.40 kPa depending on the UVB exposed dose (0-2160 mJ/cm²). Therefore, UVB radiation showed a dose-dependent inhibitory effect on cell viability, proliferation rate and Youngs modulus.

Abstract: New strategies to make cultured fibroblasts grafts more appealing are aimed at reducing the time spent in culture and improving the handling and biologic properties. In the present study, we developed a simple and effective method to fabricate dermal fibroblasts-populated membrane based on (1) the use of fibrin as a 3-dimensional matrix and (2) the use of cell- mediate contraction to make a self assembled, detachable cells-populated membrane. Human dermal fibroblasts were cultured by explants method. The fibroblasts encapsulated in fibrin were transferred into 6-well culture plates which pretreated with Sigmacoat® to prevent cell binding on surface of culture dish. Fibroblasts populated fibrin matrix (FPFM) was cultured in attached condition for 7 days and in free floating condition for 1 day. The FPFM were contracted, spontaneously released from culture plate, compacted, and formed tissue-like membrane. The fabricated FPFM revealed uniformly distributed cells and newly synthesized extracellular matrix was deposited in matrix. FPFM could successfully graft into full-thickness cutaneous defect of nude mice, and showed significantly increased wound closure rate. Our results demonstrate that the FPFM membrane delivery system allows for restoration of both the epidermal and dermal compartments.

Abstract: Three-dimensional gelatin-chondroitin 6 sulphate-hyanuronic acid biomatrix was used as the scaffold to investigate the phenotypic and molecular expression in human keratinocytes (K) and dermal fibroblasts (FB) in three different culture conditions in vitro. The cells were cultured in either monolayer (K or FB only) or coculture (K&FB) model. The deposition of basement membrane proteins secreted by these two kinds of cells was quantitatively characterized by real-time PCR. In the results, dermal fibroblasts were shown to synthesize and deposit laminin 5, type IV and type VII collagen, whereas keratinocytes produced integrin alpha 6 and beta 4 as well as laminin 5 and collagen type IV, VII. Interestingly, the integrin beta 4 subunit was not expressed either in keratinocytes or dermal fibroblasts monoculture but was seen in organotypic coculture model in the early culture period. Furthermore, we found that the expression of those marker compounds was reciprocally regulated when keratinocytes and dermal fibroblasts were cultured together. These results indicated that keratinocyes and dermal fibroblasts worked together to reconstruct dermal-epidermal basement membrane (BM) zone. In brief, our data provide the first time in directly quantifying the expression of BM proteins by using real-time PCR, and also demonstrate that BM proteins were regulated by cell-cell interaction.