含bFGF和Vit C的载体复合膜负载骨髓间充质干细胞修复兔创面的实验研究

Experimental Study on Repair of Rabbit Wound by Carrier Complex Membrane with bFGF and Vit C Loaded Marrow Stem Cell

目的含bFGF和Vit C的载体复合膜负载骨髓间充质干细胞植入兔创面,探讨其对真皮新生、重建速度的影响和促进表皮修复速度的最佳阶段。方法抽取24只新西兰大耳白兔的骨髓间充质干细胞,体外分离、培养。每只兔的背部脊柱两侧做3个创面,深度直达深筋膜,面积约为3.14cm2,随机分为A、B、C三组。A组创面置入含有MSCs、bFGF和Vit C的羊膜载体复合膜。B组创面置入只含有MSCs的羊膜载体复合膜。C组创面只置入羊膜载体复合膜。计算出创面愈合率,采用大体观察、常规组织学观察(HE染色、Masson染色、Van Giesonr染色)、Ⅰ型胶原免疫组织化学观察、墨汁灌注法等化学染色动态观察创面愈合情况。结果A组在7d、14d、21d的新生真皮明显较B组相应时间点的厚度、活跃的Ⅰ型胶原阳性细胞数量、血管和胶原纤维均较B组的多。B组比C组的修复效果好。A组术后14d和21d新生真皮表面,表皮再生长入并覆盖真皮修复创面的速度均比B组的快;B组的表皮覆盖速度又比C组的快。对创面愈合率进行统计学处理,A组比B组好,B组比C组好,差异有统计学意义。结论羊膜载体复合膜植入全层皮肤缺损后,添加MSC、bFGF和Vit C等因素,能加速真皮的修复和重建,并且在真皮修复过程中可能有表皮新生和重建的最佳时期。

Objective Carrier Complex Membrane with bFGF and Vit C loaded marrow stem cell was transplanted on the deep-partial thickness wounds. To explore the epidermis renascence and regenerate in the process of the dermis restore. Methods MSCs were isolated from 24 rabbit＇s marrow. The MSCs were cultured and purified in vitro. Three deep-partial thickness wounds were produced on the back of each of the rabbit. Each one was divided into A, B and C group randomly. A group was the experimental group which consisted of MSCs, cattle amniotic membrane gelatin chitosan, bFGF and Vit C. B group consisted of MSCs, cattle amniotic membrane gelatin chitosan. C group consisted of cattle amniotic membrane gelatin chitosan. Cell morphology changes, Immunohistochemical examinations and histological examination were performed to evaluate the velocity and the quality of the wound healing after transplanting, HE dyeing after put ink perfusion into blood vessel. Results The depth of the renascence dermis, the number of the active type I collagen masculine cell, the number of the blood vessel, all these A group are better than B group and C group. On the renascence dermis exterior there are lots of regenerate epidermis from the around normal skin, A group was better than B group and C group. Conclusion Amnion carrier compound membrane was transplanted to deep-partial thickness wounds append with MSC bFGF and Vit C can influence the rapidity renascence and regenerate of the dermis.