组织工程技术修复全层皮肤缺损的研究

Repairment of full-thickness wounds utilizing tissue engineering techniques

目的 探讨用异体脱细胞真皮基质 (ADM )复合自体表皮角朊细胞 (AKCs)移植修复全层皮肤缺损的可行性。方法 用DispaseII/TritonX 10 0制备ADM。在猪背部两侧分别作 5cm× 5cm的全层皮肤缺损 ,用ADM覆盖创面。 7～ 10d后 ,将AKCs与Pluronic混匀涂布于一侧ADM表面 ,另一侧单纯ADM组作为对照组。术后观察愈合情况 ,比较创面收缩率 ,取材作组织学及电镜观察。结果 ADM去除了所有细胞成分 ,胶原束结构及排列均保持正常 ;ADM在创面成活 ,能诱导成纤维细胞长入和新生血管形成 ,并支持移植于其表面的AKCs生长。 4周时 ,对照组收缩至原面积的 41% ,实验组为原面积的 65 %。结论 ADM可以作为重建真皮层的诱导支架 ;ADM与AKCs复合移植可以形成新生皮肤并修复全层皮肤缺损。

Objective To explore the possibility of composite transplatation of acellular allo dermal matrix (ADM) and cultured autologous keratinocytes (AKCs) to repair the full thickness skin defects in a porcine model.Methods ADM was prepared from allogeneic skin by treated with Dispase Ⅱ and Triton X 100.Two full thickness skin defects in size of 25 cm 2 were created on the dorsum of each Changfeng crossbred pig,and both wounds were treated with a same size ADM.7 10 days later,cultured AKCs in 30% pluronic F 127 were applied on the top of the survived ADM and the ADM on the other side was left untreated as control.The wound areas were measured to determine the rate of ADM contraction.Biopsy of ADM was performed after transplantation for histology and electron microscopic examinations.Results ADM retains structural integrity.Histological analysis revealed normal collagen bundle and the absence of cellular elements;ADM could survive on full thickness wounds.Histological analysis revealed that ADM supported fibroblast infiltration and neovascularization;ADM could support keratinocyte growth.At 28 days post grafting,contraction of ADM in control group was 41% and 65% in experimental group.Conclusion Dispase Triton treated dermal matrix can act as a template for reconstituting viable dermis;Neo skin could be formed in vivo by composite transplantation of ADM and cultured autologous keratinocytes.