鼠胚胎成纤维细胞三维培养修复大鼠全层皮肤缺损效果及其机制.docVIP

鼠胚胎成纤维细胞三维培养修复大鼠全层皮肤缺损效果及其机制 　　[摘要] 目的 观察鼠胚胎成纤维细胞三维培养修复大鼠全层皮肤缺损的效果，并对其机制进行分析。 方法 采用组织块法培养鼠胚胎成纤维细胞，将鼠尾胶原溶液和以血清重悬的鼠胚胎成纤维细胞溶液混合后形成凝胶构建组织工程皮肤，移植于大鼠背部全层皮肤缺损创面。实验组30只全层皮肤缺损大鼠采用组织工程皮肤进行修复。对照组30只全层皮肤缺损大鼠不做任何处理。两组创面分别进行HE染色和免疫组织化学染色，计算微血管密度，观察其愈合过程。 结果 术后20 d实验组用组织工程皮肤覆盖的大鼠背部创面已完全愈合，新生上皮外观与结构和正常皮肤相似；对照组创面仍有痂皮覆盖，去除痂皮可见肉芽组织红润致密，创面周围新生上皮呈粉红色，边缘锐利皱缩。实验组创面组织中CD31表达及微血管密度均明显高于对照组，差异有统计学意义（P 0.05）。 结论 鼠胚胎成纤维细胞三维培养对动物皮肤全层缺损的修复有很好的效果，在修复皮肤损伤方面具有良好的临床应用前景。 　　[关键词] 胚胎成纤维细胞；三维培养；皮肤缺损；微血管密度 　　[中图分类号] R641 [文献标识码] A [文章编号] 1674-4721（2016）10（c）-0012-04 　　[Abstract] Objective To investigate the effect and mechanism of three dimensional culture of rat embryonic fibroblasts in the treatment of full-thickness skin defect in rats. Methods Full-thickness skin wound was created on the back of rats. Tissue block method was used to culture rat embryonic fibroblast cells. The rat embryonic fibroblast cells resuspended in serum were mixed with tail collagen solution to structure tissue engineered skin. In the experimental group， full thickness skin defect rats （n=30） were transplanted with tissue engineered skin in the wound area. In the control group， full-thickness skin defect rats （n=30） were not given any special treatment. The microvascular density was calculated and the healing process was observed by HE staining and immunohistochemistry method. Results 20 d after operation， in the experimental group the wound areas transplanted with tissue engineering skin on the back of rat were completely healed； the appearances and structures of newborn epithelium were similar to the normal skin. The wounds of the control group did not show healing and were covered with scar； removing scab skin visibled granulation tissues were ruddy and compact； there were pink wounds around new epithelium， and shrinkage around tissue edges. In the experimental group， and the expression of CD31 and microvessel density in wound tissues were significantly higher than those in the control group （P 0.05）. Conclusion Thr