表皮干细胞复合羊膜构建组织工程皮肤修复糖尿病难愈创面

目的探讨表皮干细胞复合羊膜构建组织工程皮肤修复糖尿病难愈创面的可行性和方法。方法分离培养SD大鼠皮肤表皮干细胞(ESCs),并用5-溴脱氧尿嘧啶(BrdU)标记。建立糖尿病SD大鼠创面模型,随机分为A、B、C 3组,A组创面移植羊膜负载BrdU标记的ESCs组织工程皮肤;B组创面移植羊膜;C组为空白对照组,创面未给予干预。观察创面愈合情况、计算创面愈合率,HE及免疫组织化学SP法检测创面愈合组织中BrdU及增殖细胞核抗原(PCNA)的表达。用图像分析软件测量阳性细胞积分光密度平均值。结果与B、C组比较,A组创面愈合速度加快,创面愈合率明显增高,组间比较差异有显著性统计学意义(P0.01)。A组创面及新生表皮中可见BrdU阳性细胞,B、C组皮肤创面组织中始终未见BrdU阳性细胞。各组创面组织中可见PCNA阳性细胞表达,但A组的阳性细胞积分光密度值与B、C组比较差异显著(P0.01)。结论表皮干细胞复合羊膜构建组织工程皮肤可有效促进糖尿病创面愈合。

Tissue Engineered Skin with Epidermal Stem Cells and Amniotic Membrane for Diabetic Impaired Wound

Objective To investigate the feasibility of tissue engineered skins with epidermal stem cells and amniotic membrane for diabetic impaired wound.Methods Epidermal stem cells（ESCs） derived from SD rats were isolated,cultured,identified and labled with 5-bromo-2＇-deoxyuridine（BrdU） in vitro.The wound model of diabetic SD rats were established,and then divided into A,B,C group.Tissue engineered skins with labled BrdU ESCs and amniotic membrane were implanted to group A.Amniotic membrane were implanted to group B.Group C as control group received no cells or amniotic membrane.At 7,14 days after transplantation,general situation of wound healing,the healing rate of wound,hematoxylin and eosin（HE） staining,immunohistochemical staining of Brdu and proliferating cell nuclear antigen（PCNA）in wounds were investigated.Integral optical density mean of positive cells was measured with image analysis software.Results Compared with the other two groups,the healing time of group A was faster.And the wound healing rate in group A was significantly higher at different observation times（P〈0.01）.BrdU-positive cells in the wounds and newborn epidermis of group A were visible,while wound tissue of the other two groups had no BrdU-positive cells.PCNA-positive cells in wound tissue of each group could be seen,but integral optical density meanof positive cells was significantly different in group A compared to that in group B,C（P〈0.01）.Conclusion Tissue engineered skins with epidermal stem cells and amniotic membrane may contribute to healing of diabetic impaired wound.