体外构建3D人工皮肤

发布时间：2019-04-10 07:00

【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。
[Abstract]:Background: 3D printing artificial skin has been studied, but the skin epidermis layer can not differentiate into multi-layer. Objective: to optimize the construction method of artificial skin and obtain better artificial skin. Methods: collagen was extracted and purified from bovine tail and rat tail respectively. Primary fibroblasts and keratinocytes were isolated and cultured from the prepuce tissue of children, and then the fibroblasts were embedded in bovine tail (or rat tail) collagen to construct the dermis. Keratinocytes were cultured on fibroblasts containing collagen of rat tail (or bovine tail) for 7 days and then cultured in air for 7 days to obtain artificial skin. The contractility of artificial skin, water contact angle, hematoxylin-eosin staining, Sirius red staining and immunofluorescence staining were observed. Results & conclusion: (1) the dermal area of artificial skin was reduced by 50% and that of bovine tail collagen by 21%. (2) the contact angle of the artificial skin constructed by bovine tail collagen was similar to that of human skin, which was better than that of the artificial skin constructed by rat tail collagen. (3) the results of Sirius red staining showed that compared with the artificial skin constructed by rat tail collagen, the collagen refractive property of the artificial skin was stronger, the arrangement of collagen was more orderly and the fiber was thicker than that of the artificial skin constructed by rat tail collagen. (4) hematoxylin-eosin staining showed that compared with the artificial skin constructed by rat tail collagen, bovine tail collagen could form more layers in the epidermis of artificial skin. (5) the results showed that better artificial skin could be constructed by inoculating keratinocytes on fibroblasts embedded with bovine tail collagen.
【作者单位】：浙江大学化学工程与生物工程学院;
【基金】：国家自然科学基金(21476197)~~
【分类号】：R318.1

【相似文献】