组织工程皮肤自体及异体移植后表皮分化及重建的动物实验研究

Experimental Study on Epidermal Differentiation after Autograft and Allograft of Swine Tissue Engineered Skin

目的研究猪组织工程皮肤移植后表皮层角蛋白分化标志的变化情况,为组织工程皮肤临床应用提供实验依据。方法按本科已建立的方法构建猪的组织工程皮肤,进行自体及同种异体移植实验,并于移植后1周、4周、8周取标本,行组织学检查和免疫组化染色,观察移植后组织工程皮肤表皮层角蛋白的表达、表皮细胞分化和重建情况。结果猪组织工程皮肤自体及同种异体移植后均存活,创面达到理想的愈合效果;组织学检查显示表皮分化良好,表皮层广谱角蛋白抗体免疫组化染色阳性;基底层以上细胞角蛋白K10抗体免疫组化染色阳性;基底层细胞角蛋白K5抗体免疫组化染色阳性。结论构建的猪组织工程皮肤自体及同种异体移植后,可获得理想的创面愈合效果,表皮分化和再构建良好,与正常皮肤相似。本实验为组织工程皮肤临床应用提供了有力的实验依据。

Objective To provide experimental evidence for clinical application of the tissue engineered skin, a swine tissue engineered skin was fabricated to study the change of epidermal differentiation markers after autograft and allograft.Methods Serially passaged cells were used to fabricate the tissue engineered skin as we had reported.Then the tissue engineered skin were autografted and allografted onto full thickness excised wounds of two swines after organotypic culture in vitro.It’s viability and wounds healing status were observed.Biopsy of the grafts taken at week 1,4 and 8 respectively was examined histologically and immunohistochemically to investigate the reconstruction of the tissue engineered skin,expression of epidermal keratin, differentiation of the epidermis.Results The tissue engineered skin was living after autograft and allograft,merged with the surrounding normal skin of the swine.The wounds healed well.The epidermis of the tissue engineered skin transplanted onto the swine turned thick, its ridge was distinctness.The tissue engineered skin transplanted onto the swine showed immunohistochemically that broad-spectrum keratin was expressed in epidermis,cytokeratin 10(K10) in the suprabasal layer and cytokeratin 5(K5) in the basal layer.Conclusion Wounds healed well after autograft and allograft.The tissue engineered skin reconstruct well.This animal study provides experimental evidence for clinical application of the tissue engineered skin.