组织工程皮肤与正常皮肤角蛋白构型的对比研究

Comparison of keratin pattern between tissue engineered skin and normal skin

目的:对比研究培养的组织工程皮肤与正常皮肤的角蛋白构型,探讨培养后的表皮分化情况。方法:将毛乳头细胞、真皮鞘细胞或成纤维细胞分别与鼠尾胶原混合,制成间质细胞凝胶,再在细胞凝胶表面接种角质形成细胞,体外培养2周,组织切片用15种角蛋白单抗和两种整合素α单抗进行免疫组化染色。结果:在重建的表皮中,染色结果基本相同于正常表皮,抗原性相对要弱些,在正常表皮大多抗体可稀释数倍,而重建表皮中一般都用原液才染色较强。所不同的是RCK107在重建表皮中全表皮层弱阳性,RKSE60在表皮+真皮鞘细胞中基底层弱阳性反应。结论:在培养条件下重建的表皮相似于体内的组织学发生,角蛋白构型基本上相同于体内,但真皮鞘细胞还诱导了低相对分子质量角蛋白表达,毛乳头细胞阻止了K10表达,说明它们与成纤维细胞还存在着功能上的差异。

AIM:To compare the keratin pattern between the tissue engineered skin and normal skin so as to investigate the differentiation of tissue engineered epidermis.METHODS:The dermal papilla cells,dermal sheath cells or fibroblasts were mixed with rat tail collagen to prepare the interstitial cell gel,then inoculated keratinocytes on the surface of cell gel and cultured in vitro for two weeks to reform the tissue engineered skin.The tissue sections were stained with 15 anti keratin and anti integrin α monoclonal antibodies with immunohistochemical method.RESULTS:In the reconstructed epidermis,the staining results was similar to that in the normal skin,but the antigen was lower than that of normal skin,because in the normal skin sections the primary antibodies could be diluted manifold and in the reconstructed epidermis it was high in primary concentration.It was different that RCK107 was weakly positive in full layer epidermis and RKSE60 was weakly positive in the basal layer of the epidermis formed by the dermal sheath cells.CONCLUSION:Under culture in vitro,the reconstructed epidermis was similar to the skin in vivo in morphogenesis and the keratin pattern was also similar to the normal skin.The dermal sheath cells can induce the low molecular mass keratin expression and the dermal papilla cells can prevent keratin 10 expression,indicating that those cells differ from fibroblasts in function.