摘要
目的评价预血管化去细胞神经整合自体定向分化脂肪间充质干细胞修复周围神经缺损的效果。方法取大鼠腹股沟脂肪组织分离间充质干细胞，同时将血管内皮生长因子（vascular endothelial growthfactor，VEGF）纳米微囊显微注入化学去细胞神经，植入取脂肪创口内预血管化。采用神经诱导剂将脂肪间充质干细胞定向诱导分化为类雪旺细胞，2周后植入预血管化的去细胞神经修复自体大鼠15mm坐骨神经缺损为A组（实验组）；去细胞神经植入体内预血管化移植自体诱导分化的脂肪间充质干细胞，但不加VEGF纳米微囊为B组（对照组）；化学去细胞神经移植自体诱导分化的脂肪干间充质细胞为C组；自体神经移植组为D组；VEGF纳米微囊预血管化组为E组。术后第7天取A、B、C组移植神经，检测微血管密度，并用TUNEL法检测细胞凋亡情况。术后12周测各组肌电图、小腿三头肌湿重恢复率以及轴突计数评价其修复周围神经缺损的效果。结果术后第7天C组大部分细胞凋亡，凋亡率最高，A组细胞凋亡率低于B组，微血管密度A组〉B组〉C组。术后12周，A组神经传导速度、动作电位波幅，小腿三头肌湿重恢复率以及有髓神经轴突数量高于B、C、E组，差异有统计学意义（P〈0．05）。结论去细胞神经预血管化能提高移植细胞存活率，具有明显的促进神经再生作用。
Objective To evaluate the effectiveness of prevascularized acellular nerve graft combined with autologous differentiated adipose-derived stem cells on repairing nerve defect in rats. Methods Adiposederived stem cells （ADSCs） were isolated from rat inguinal fat pad. Vascular endothelial growth factor （VEGF） encapsulated nanoparticles were injected into the chemically decellularized nerve graft which in turn was buried into the incision for fat harvesting. The rat was kept for 2 weeks to allow prevascularization of the acellular nerve graft. The ADSCs were cultured under neurogeneic conditions for 2 weeks to differentiate into Schwann cells. A 15 mm sciatic nerve gap in rats was repaired by VEGF nanoparticle prevascularized acellular nerve graft injected with differentiated ADSCs （Group A）, by non-VEGF nanoparticle prevascularized acellular nerve injected with differentiated ADSCs （Group B）, by acellular nerve injected with differentiated ADSCs （Group C）, by autologous nerve graft （Group D）, and by VEGF nanoparticle prevascularized acellular nerve graft （Group E）. On postoperative day 7 grafts from Groups A, B and C were harvested. Microvessel density （MVD） and graft cell apoptosis were examined. After 12 weeks, nerve conduction study, triceps sm＇ae muscle wet weight, and neuromorphometry analysis were carried out to evaluate the extent of nerve regeneration. Results Seven days postoperatively, graft cell apoptosis was most prominent in group C. Apoptosis in group A was less severe than Group B. MVD was highest in group A, followed by group B and then group C. Evaluations at 12 weeks postoperatively showed significantly higher nerve conduction velocity, larger action potential amplitude, higher triceps surae muscle weight recovery rate and larger myelinated nerve fiber count in Group A than in Group B, Group C and Group E （ P 〈 0.05 ）. Conclusion There is a synergistic effect of acellular nerve and differentiated ADSCs in repairing nerve gap in rats. The prevascularized acell
出处
《中华手外科杂志》
CSCD
北大核心
2014年第3期216-219,共4页
Chinese Journal of Hand Surgery
基金
浙江省自然科学基金（Y2100253）
温州市科技计划项目（Y20130203）
关键词
神经再生
干细胞
间充质
预血管化
去细胞神经
Nerve regeneration
Stem cells, mesenchymal
Prevascularization
Acellular nerve
