异体骨髓间充质干细胞联合自体PRP促进皮肤慢性溃疡创面愈合的实验研究

Combination of bone marrow-derived stem cells and autologous platelet-rich plasma promote wound healing of chronic skin ulcer in rats

目的探讨在皮肤慢性溃疡处注射异体骨髓间充质干细胞（BMSC）和自体富血小板血浆（PRP）对皮肤的再生和重建的作用。方法选择健康6—8周龄SD大鼠17只，取其中1只大鼠，采用全骨髓法体外分离培养BMSC，取传至第3代细胞，移植前48h行DiI荧光标记。剩余16只大鼠麻醉后制作全层皮肤缺损模型，于其背部脊柱两侧制作4个全层皮肤缺损创面并随机分为4组。A组局部注射异体BMSC悬液联合PRP治疗，B组局部单纯注射异体BMSC悬液，C组局部单纯注射PRP治疗，D组局部注射等量生理盐水。治疗7、14d采集标本观察结果。采用大体观察、苏木素一伊红染色、荧光显微镜观察创面愈合情况。结果体外分离培养的BMSC具有成脂、成骨、成神经分化潜能。在治疗后7、14d，A、B、C组创面愈合时间均短于D组，A组愈合时间比B、C组缩短。组织学观察发现A组新生真皮相应时间点的厚度、血管和胶原纤维均明显多于其他3组。结论在皮肤慢性溃疡处联合应用异体BMSC和自体PRP，能加速真皮的修复和重建。

Objective To investigate the effects of local injection with exogenous bone marrow mesenchymal stem cells （BMSCs） and antologous platelet-rich plasma on skin regeneration and reconstruction in rats with chronic skin ulcer. Methods Seventeen SD rats aged 6-8 weeks were used in this study. BMSCs were isolated from bone marrow of one rat. The 3rd passage of BMSCs was DiI-labeled 48 hours before transplantation. Full thickness skin defect model with four wounds on the back was established in the remaining 16 rats after anesthesia. Four wounds of the rats were randomly divided into 4 groups. Skins wounds were locally injected with BMSCs and PRP in group A, BMSCs in group B, PRP in group C, and physiological saline solution in group D. Histological analysis was performed to evaluate the velocity and the quality of the wound. Fluo- rescence microscope was used to evaluate the wound healing 7 days and 14 days after treatment. Results BMSCs were capable of adipogenic, osteogenic and chondrogenetic differentiation in vitro. On day 7 and day 14, wound healing times of group A, B, C were significantly shorter than that of group D. And wound healing time of group A was shorter than those of group B and C. Histological analysis showed that the depth of the renascence dermis, the number of the blood vessel and collagen fiberall in group A were all better than those in any other groups. Conclusions In chronic skin ulcers, combined local application of exogenous BMSCs and autologous PRP can accelerate the wound repair and reconstruction of the dermis.