Effect of Oligomeric procyanidins of Vitis vinifera on the wound healing in the Sprague-Dawley rat

Abstract

Purpose: The purpose of this study is to evaluate the effect of OPC on diverse steps of wound healing, especially on the migration, cytokines related to angiogenesis, and collagen synthesis. In addition, we investigate the effects of OPC on rat skin wound model. Materials and Methods: Immortalized epidermal keratinocyte cell line (HaCaT cells) and human neonatal dermal fibroblasts (HNDFs) were used in this study. Viability of the HaCaT cells and HNDFs in response to OPC was determined by the XTT assay. Scratch wound migration assays of the HaCaT cells and HNDFs were performed. Western blot analysis of VEGF and thrombospondin-1 (TSP1) in a HaCaT cells and HNDFs was performed for the evaluation of angiogenesis. Western blot analysis of procollagen in HNDFs was also done. In twenty male Sprague-Dawley rats, the effect of OPC in wound healing was evaluated. Five and ten days after surgery, biopsy was harvested and Hematoxylin & eosin stain (H&E stain), Trichrome Masson stain, immunohistochemistry of VEGF and TSP1 were performed Results: There were no significant differences in the viabilities of the HaCaT cells and HNDFs under these various concentrations of OPC. Migration of HaCaT cells and HNDFs was inhibited by OPC in a dose-dependent manner. OPC could inhibit VEGF expression and increase TSP1 expression in HaCaT cells. OPC seems to inhibit VEGF and TSP1 expression in HNDFs. OPC could inhibit collagen synthesis of HNDFs significantly. Wounds in the experimental groups healed slower than the control group, especially in 25, 50 mg/ml OPC-treated group. More complete and mature epithelialization of the epidermis could be observed in control group in H&E stain. The Trichrome Masson stain of the experimental group showed less copious and horizontally-oriented collagen lay-down compared to the control group. VEGF expression decreased and TSP1 expression increased in immunohistochemical stains of the experimental group compared with control group. Conclusion: OPC from vitis vinifera seed inhibit the wound healing process through inhibition of fibroblasts and keratinocytes migration, angiogenesis, and collagen synthesis of rat skin wound.