脂肪干细胞和纤维蛋白胶快速构建人工皮肤修复创面的研究

目的:探讨绿色荧光蛋白(GFP)转基因小鼠脂肪组织来源干细胞(ADSCs)与纤维蛋白胶复合快速成型构建组织工程皮肤替代物修复皮肤缺损的可行性.方法:采用胶原酶消化GFP小鼠脂肪组织,获得GFP小鼠脂肪干细胞(ADSCs),并证明其具有成骨和成脂的多向分化能力,经体外扩增后复合纤维蛋白胶快速成型构建组织工程活性皮肤替代物.15 只C57鼠随机分成3组:ADSCs复合纤维蛋白胶组织工程活性皮肤替代物实验组、单纯纤维蛋白胶移植组和空白对照组.将各组相应材料分别移植于C57BL小鼠背部皮肤缺损处,比较观察创面修复效果.结果:分离的细胞具有不断的增殖能力,并具有多向分化潜能.同种异体移植后,免疫荧光检测创面有大量GFP小鼠脂肪干细胞存活.ADSCs复合纤维蛋白胶可以缩短创面愈合时间.组织学观察显示,ADSCs复合纤维蛋白胶实验组上皮形成较厚,真皮层胶原纤维排列有序,成纤维细胞数量较单纯纤维蛋白胶移植组和空白对照组明显增多.结论:GFP-ADSCs复合纤维蛋白胶构建的组织工程皮肤替代物移植后可加速小鼠皮肤创面愈合,提示以ADSCs作为种子细胞复合纤维蛋白胶快速成型构建的组织工程皮肤可用于皮肤缺损的修复治疗.

Rapid construction of engineered skin substitutes by adipose tissue-derived stem cells and fibrin glue for full thickness skin wound repair

To investigate the feasibility of construction of engineered skin substitutes with adipose tissue derived stem cells ( ADSCs)from GFP-mouse and fibrin glue. Methods: Adipose tissues were isolated from GFP-C57BL mouse groin, the fat was cut into pieces and incubated in a collagenase solution to establish ADSCs culture. The differentiation ability into adipocytes and osteoblasts of the ADSCs were investigated. The amplified cells in vitro were seeded in fibrin glue to form tissue engineered skin substitutes. Fifteen C57BL mice were randomly divided into three groups; experimental group was treated with ADSCs-fibrin glue tissue engineered active skin substitutes. Single fibrin glue and blank were served as two control groups. The materials were grafted onto total skin defects on the back of mice. The effect of wound repair was observed histologically. Results; The cultured ADSCs grew well in vitro. The cells showed characteristics of multipotent adult stem cells. ADSCs-fibrin glue shortened the wound healing time. Histological observation showed that thicker epithelium was formed and collagen fibers arranged in order in the ADSCs-fibrin glue experimental group,the number of fibroblasts was significantly increased as compared with those in single fibrin glue group or in the blank control group. Conclusion : GFP- ADSCs combined with fibrin glue can accelerate the mouse skin wound healing. It suggests that ADSCs with fibrin glue maybe an optional method of engineered skin reconstruction for wound repair.