CAJ | 학술논문

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(vascularendothelial growth factor,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15 mm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.
목적 평개예혈관화거세포신경정합자체정향분화지방간충질간세포수복주위신경결손적효과.방법 취대서복고구지방조직분리간충질간세포,동시장혈관내피생장인자(vascularendothelial growth factor,VEGF)납미미낭현미주입화학거세포신경,식입취지방창구내예혈관화.채용신경유도제장지방간충질간세포정향유도분화위류설왕세포,2주후식입예혈관화적거세포신경수복자체대서15 mm좌골신경결손위A조(실험조);거세포신경식입체내예혈관화이식자체유도분화적지방간충질간세포,단불가VEGF납미미낭위B조(대조조);화학거세포신경이식자체유도분화적지방간간충질세포위C조;자체신경이식조위D조;VEGF납미미낭예혈관화조위E조.술후제7천취A、B、C조이식신경,검측미혈관밀도,병용TUNEL법검측세포조망정황.술후12주측각조기전도、소퇴삼두기습중회복솔이급축돌계수평개기수복주위신경결손적효과.결과 술후제7천C조대부분세포조망,조망솔최고,A조세포조망솔저우B조,미혈관밀도A조＞B조＞C조.술후12주,A조신경전도속도、동작전위파폭,소퇴삼두기습중회복솔이급유수신경축돌수량고우B、C、E조,차이유통계학의의(P＜0.05).결론 거세포신경예혈관화능제고이식세포존활솔,구유명현적촉진신경재생작용.
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 surae 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 acellular nerve improves survival of transplanted cells,which is benefit for nerve regeneration.