| Objective:The purpose of this manuscript is to evaluate the effects of Panax notoginseng saponin Rgl on HUVEC proliferation, migration and VEGF expression in vitro and to develop a collagen/chitosan-gelatin microspheres scaffold controlled release Rgl and to evaluate its effect on tissue angiogenesis and wound regeneration in vivo.Methods:The consistent and steady release of angiogenic factors is an important factor in the promotion of angiogenesis in dermal substitutes, which usually lack, yet need, a vascular network. Rgl, isolated from Panax notoginseng (PNS) saponins, has been proven to be a candidate drug for angiogenesis, whereas its applications in tissue engineering have been rarely reported. In this study, the effect of Rgl on the biological behavior of human umbilical vein endothelial cells (HUVECs) was investigated in vitro. Furthermore, to develop a local delivery and subsequent controlled release system, PNS Rgl was first introduced into gelatin microspheres (GMS), which serve as drug carriers, and then incorporated into collagen/chitosan scaffolds to fabricate a collagen/chitosan-GMS (CC-GMS) scaffold. The morphology of the CC-GMS scaffold was investigated in vitro. To characterize the effect of the CC-GMS scaffold on angiogenesis and tissue regeneration, the CC-GMS scaffold was subcutaneously embedded in Sprague-Dawley (SD) rats. At days7,10,14, and21after implantation, the implants were imaged to analyze the newly formed blood vessels, and tissue specimens were harvested for histology, immunofluorescence, real-time quantitative PCR (RT-qPCR), and western blotting analyses.Conclusions:The results demonstrated that Rgl promoted the proliferation of HUVECs in a time-and dose-dependent manner. Moreover, the CC-GMS scaffold showed more rapid angiogenesis than did the control groups in vivo. Our data suggested that Rgl could be a candidate for accelerating angiogenesis in skin tissue engineering, and the ability of the CC-GMS scaffold to promote angiogenesis demonstrates its potential in skin tissue engineering. |
