角质细胞与成纤维细胞混合移植修复裸鼠全层皮肤缺损的初步报告

Mixed keratinocytes and fibroblasts fibrin glue suspension for coverage of full thickness skin wound in nude mice: a preliminary report

目的 利用组织工程原理探讨修复全层皮肤缺损的理想方法。方法 以裸鼠为动物模型 ,在皮肤全层缺损区域分别移植纤维蛋白胶 (n =10 ) ,纤维蛋白胶角质细胞悬液 (n =10 ) ,纤维蛋白胶成纤维细胞悬液 (n =10 )以及纤维蛋白胶角质细胞成纤维细胞悬液 (n =10 ) ,术后每天对伤口进行大体观察 ,第 5 ,7,10 ,14,2 1,35d ,分别取材活检行组织学及免疫组织化学检查。结果 移植有角质细胞组 (2和 4组 )的创面愈合快 ,术后 10d组织学提示创面完全上皮化 ,抗人特异性HLA 1型抗原、抗involucrin染色和抗Ⅶ型胶原染色阳性证明新生上皮由移植的人角质细胞形成 ,抗involucrin染色阳性又证明角质细胞分化成熟有角质层形成 ,抗Laminin染色、抗Ⅶ型胶原染色阳性提示早期基底膜形成。组织学检查提示第 4组新生上皮有许多类似皮钉样结构。结论 培养的角质细胞 ,成纤维细胞结合纤维蛋白胶移植到创面上后 ,可以形成复层分化良好。

Objective To investigate an ideal approach of skin repair using skin tissue engineering principle. Methods\ In nude mice, full thickness skin wound was covered either with fibrin glue, or fibrin glue keratinocytes and fibroblasts fibrin glue suspension. Results\ Groups with keratinocytes (group II and IV) exhibited accelerated wound healing. On day 10 after grafting the wounds were completely epithelialized, and the neoepithelia were regenerated from grafted human keratinocytes, which was demonstrated by positive anti human HLA I staining, anti human involucrin staining and anti human collagen type VII staining. Positive anti involucrin staining indicated the terminal differentiation of grafted keratinocytes. The early reformed basement membrane was indicated by anti laminin and collagen type VII staining. Many rete ridges were observed only in group IV. Conclusion\ Cultured keratinocytes and fibroblasts when suspended in fibrin glue and grafted onto a wound can form a multilayer, well differentiated new skin tissue similar to normal structure and function.