摘要
背景:已有研究采用3D打印人工皮肤,但存在皮肤表皮层不能分化为多层等缺陷。目的:优化人工皮肤的构建方法,获得更好的人工皮肤。方法:从牛尾和鼠尾中分别提取与纯化胶原。从儿童包皮组织中分离培养原代成纤维细胞和角质细胞,再以牛尾(或鼠尾)胶原包埋成纤维细胞,构建真皮层;将角质细胞接种于含鼠尾(或牛尾)胶原包埋的成纤维细胞上培养7 d,再气提培养7 d,获得人工皮肤。观察人工皮肤的收缩性、水接触角,并进行苏木精-伊红染色、天狼猩红染色及免疫荧光染色。结果与结论:(1)鼠尾胶原构建人工皮肤真皮层面积下降了50%,牛尾胶原构建人工皮肤真皮层面积只下降了21%;(2)牛尾胶原构建人工皮肤的接触角与人皮肤接近,比鼠尾胶原构建人工皮肤的疏水性更好;(3)天狼猩红染色结果显示,与鼠尾胶原构建的人工皮肤相比,牛尾胶原构建人工皮肤的胶原折光性更强,胶原排列更有序,纤维更粗;(4)苏木精-伊红染色显示,与鼠尾胶原构建人工皮肤相比,牛尾胶原构建人工皮肤真皮层的表皮可形成更多层;(5)结果表明,将角质细胞接种于牛尾胶原包埋的成纤维细胞上,可构建更好的人工皮肤。
BACKGROUND： Epidermis is unable to differentiate into stratum corneum and other parts in the previous artificial skin constructed using three-dimensional printing. OBJECTIVE： To optimize the method of artificial skin construction to obtain more feasible artificial skin. METHODS： Type I collagen extracted from rat-tail and bovine tail was purified. Primary dermal fibroblasts and keratinocytes were isolated from children＇s foreskin, and then embedded in bovine or rat tail collage to construct the dermis; keratinocytes were seeded on the dermis for 7 days, followed by 7-day air liquid interface, and the artificial skin was finally achieved. The contraction and hydrophobicity by water contact angle were detected, as well as the morphology was observed by hematoxylin-eosin staining, sirius red staining and immunofluorescence analysis. RESULTS AND CONCLUSION： Seven days after fibroblasts embedded in collagen, the area of bovine collagen was reduced by 21%, while that of rat tail collagen decreased about a half. The water contact angle of the skin constructed by bovine tail collagen was similar to that of human skin, which was much higher than that of the skin constructed by rat tail collagen. Sirius red staining found that the skin constructed by bovine tail collagen had stronger refractivity, more intact structure and thicker fibers than those of the skin constructed by rat tail collagen. Moreover, more multilayer keratinocytes appeared in the skin constructed by bovine tail collagen through hematoxylin-eosin staining. In conclusion, bovine tail collagen is more available for the artificial skin construction.
出处
《中国组织工程研究》
CAS
北大核心
2016年第46期6915-6921,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(21476197)~~
