Effects of human epidermal growth factor gene-transfected mesenchymal stem cells on fibroblast migration and proliferation

人表皮生长因子基因转染间充质干细胞对成纤维细胞迁移和增殖的影响

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Abstract

OBJECTIVES: We were interested in determining whether epidermal growth factor gene-transfected mesenchymal stem cells (EGF-MSC) would accelerate fibroblast migration and proliferation. MATERIALS AND METHODS: Fibroblasts were cultured in serum-free conditioned media from EGF-MSC; RT-PCR was performed to detect expression of EGF gene in EGF-MSCs. EGF protein levels in cell culture supernatants from EGF-MSC were assayed by ELISA and proliferation of EGF-MSC-treated fibroblasts was performed using MTT assay. Effects of EGF-MSC on fibroblast migration were evaluated using scratch wound and transmigration assays. Cell adhesion molecules, cell dynamics molecules and phospho-(Ser) kinase substrate expressions of EGF-MSC-treated fibroblasts were evaluated by western blotting. RESULTS: EGF gene expression increased in EGF-MSCs and viability of EGF-MSC-treated fibroblasts was elevated. EGF-MSC-treated fibroblasts showed increased migration compared to controls. Expressions of cell adhesion molecules (β-catenin, N-cadherin), cell dynamics molecules (cofilin, ezrin) and phospho-(Ser) kinase substrates (phospho-MAPK/CDK substrate, phospho-Arg-(Ser)-X-Tyr/Phe-X-pSer motif) increased in EGF-MSC-treated fibroblasts. These results imply that EGF-MSCs contributed to enhancing the wound healing process by increased cell adhesion, dynamic effects, fibroblast migration, and proliferation. CONCLUSIONS: This study indicates that EGF-MSCs had a positive influence on fibroblast migration and proliferation and EGF-MSC may provide a useful strategy for wound healing.

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