羊膜上皮细胞胶原海绵复合体促大鼠预构皮瓣成活及其再血管化
王颖倩, 王德伟, 许家军, 李玲巧, 周虹, 卫振邦, 王天昊
中国临床解剖学杂志 ›› 2012, Vol. 30 ›› Issue (5) : 533-538.
羊膜上皮细胞胶原海绵复合体促大鼠预构皮瓣成活及其再血管化
Complex of collagen sponge seeded rat amniotic epithelial cells promotes survival and revascularization of rat prefabricated flap
目的 移植羊膜上皮细胞(AECs) 胶原海绵复合体促进预构皮瓣成活。 方法 第3代SD大鼠AECs、软骨细胞株分别植于胶原蛋白海绵支架，并提取AECs组上清液行促血管形成因子VEGF、TGFβ1、bFGF的ELISA检测。SD大鼠分正常对照组、空白对照组、软骨细胞组和AECs组。一期手术后第7、14 d取皮瓣组织行促血管形成因子的ELISA检测。二期手术后第7 d用计算机图像分析系统测定皮瓣存活率，取存活部位组织行HE染色、vWF免疫组织化学染色以分析微血管密度。 结果 AECs复合体上清液中的促血管形成因子分泌逐渐增多；组织内各因子含量以AECs组最高，AECs组皮瓣存活率明显高于空白对照组和软骨细胞组。AECs组皮肤微血管面积比、vWF因子阳性区面积比明显高于其他组。 结论 植入AECs胶原海绵复合体可促进预构皮瓣的存活，与其分泌的一些促血管形成因子促进预构皮瓣再血管化有关。
Objective To investigate the effects of amniotic epithelial cells/collagen sponge complex on promoting survival of rat prefabricated flap. Methods The 3rd passage of SD rat amniotic epithelial cells (AECs) or chondrocyte line was seeded into the collagen sponge scaffold. ELISA was taken to detect angiogenesis factors VEGF, TGFβ1, bFGF in the supernatant fluid of the AECs complex. SD rats were divided into normal control group, blank control group, chondrocyte group and AECs group. Angiogenesis factors in the flaps of each group were detected at the day 7 and 14 after the first-stage surgery. The survival of the flaps and microvasculature density were compared and evaluated by HE staining, vWF immunohistochemistry and digital image analysis system respectively one week after the second-stage surgery. Results The level of VEGF, TGFβ1 and bFGF in the supernatant fluid of the AECs complex increased gradually, and the highest concentration was detected in flap tissue of AECs group. The survival rate of the flap in the AECs group was higher than that in the blank control group and chondrocyte group. Both subcutaneous microvascular area ratio and vWF positive area ratio in the AECs group were higher than those in the other groups. Conclusions AECs/collagen sponge complex can increase the survival of prefabricated flap evidently. Angiogenesis factors secreted by the AECs promote the revascularization in the prefabricated flap.
羊膜上皮细胞 / 胶原海绵 / 预构皮瓣 / 再血管化 / 促血管形成因子
Amniotic epithelial cells / Collagen sponge / Prefabricated flap / Revascularization / Angiogenesis factors
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