微粒皮复合人工真皮构建的组织工程皮肤用于大鼠巨大创面修复的研究

Tissue Engineering Skin Made of Microskin Combined with Artificial Dermis in the Application of Huge Wound Healing in Rats

目的:评价微粒皮复合人工真皮构建的组织工程皮肤用于修复大鼠背部巨大创面的效果。方法:选取32只8周龄SD大鼠,制备鼠背巨大创面模型,并随机分为空白对照组(A组)、人工真皮组(B组)、微粒皮组(C组)及组织工程皮肤组(D组)。B、C、D组大鼠分别以打孔拉网的人工真皮、自体原位微粒皮移植物、自体原位微粒皮复合人工真皮构建的组织工程皮肤移植物修复创面,A组创面仅覆盖油纱布。观察术后第1、3、5周时的创面愈合情况;通过CK15及CD29免疫组织化学染色法观察表皮干细胞(epidermal stem cells,ESCs)在创面愈合过程中的作用;对鼠背组织切片进行HE染色、Masson染色后镜下观察,并结合扫描电镜下观察,了解创面真皮层的愈合质量。结果:D组大鼠背部创面的修复效果最佳,术后第5周时创面已基本愈合。免疫组织化学染色结果显示,D组大鼠ESCs逐渐向表皮层迁移并参与创面的愈合;D组真皮层的厚度大于其余3组(P<0.01),D组真皮层胶原纤维排列致密整齐。结论:微粒皮复合人工真皮构建的组织工程皮肤用于创面修复的效果优于微粒皮及人工真皮。

Objective：To evaluate the effect of tissue engineering skin（TES） ,which was made of microskin combined with artificial dermis ,in the healing of huge wound on the back of SD rats .Methods：The models of huge wound on the back were made from the selected 32 SD rats at 8 weeks old .They were randomly divided into blank control group （Group A） ,artificial dermis group （Group B） ,microskin group （Group C） and TES group （Group D） .The wounds in Group B ,Group C and Group D were healed by artificial dermis with punching net ,in‐situ microskin autografts ,the TES made of autogenous in‐situ microskin combined with artificial dermis , respectively . The wounds in Group A were covered with vaseline gauze . The situations of wound healing at the 1st ,3rd and 5th week after operation were observed .The effects of ESCs during the wound healing process were observed by the CK15 and CD29 immunohistochemical stalning .The healing quality of dermis on wound was evaluated by HE stalning and Masson stalning of tissue biopsies of the back of rats and observation under scanning electronic microscopy .Results：The best healing effect was in Group D and the wounds were almost healed at the 5th week after grafting .The results of immunohistochemical stalning showed that ESCs of rats in Group D had moved to the epidermal layer gradually and took part in the wound healing .The thickness of dermal layer in Group D was larger than that in the other 3 groups（P〈0 .01） and the collagen fibers in dermal layer were arranged compactly and neatly in Group D .Conclusions：The effect of the TES made of microskin combined with artificial dermis in the application of wound healing is superior to that of microskin and artificial dermis .