BMSCs-SCs共培养体系联合异种神经支架修复大鼠坐骨神经缺损的研究

BMSCs-SCs co-culture system combined acellular nerve xenograft to repairsciatic nerve defects in rats

目的探讨骨髓基质细胞(BMSCs)与雪旺细胞(SCs)共同培养联合异种神经移植体(ANX)修复大鼠坐骨神经缺损的效果。方法取SD大鼠左侧坐骨神经分离,培养SCs,4周龄雄性SD大鼠取双侧股骨及胫骨骨髓分离纯化培养BMSCs。将BMSCs与不同浓度的SCs应用Transwell培养皿联合培养,利用CCK-8检测BMSCs增殖活性。将不同分组细胞打入制备的异种神经支架内。动物实验中,选取体重(100±20)g SD雄性大鼠,在无菌条件下制备10 mm坐骨神经缺损的大鼠模型,将种子细胞-ANX移植至大鼠坐骨神经缺损处,分为5组:BMSCs-SCs组(1)、BMSCs-SCs组(2)、BMSCs-SCs组(3)、BMSCs组(4)和空白移植组(5)。术后8周,对各组大鼠行形态学实验检测评价移植神经再生的效果。结果通过CCK-8检测发现共培养体系中的BMSCs细胞增殖活性强于单细胞组,呈典型的极性漩涡状生长。经8周饲养后的动物实验,通过检测大鼠坐骨神经功能指数、热板实验、大鼠患侧/健侧胫骨前肌及腓肠肌湿重比率发现,共培养组坐骨神经功能恢复较好(P<0.05),坐骨神经支配的感觉方面恢复效果显著(P<0.05)。结论共培养组坐骨神经功能恢复优于单细胞组;运用共培养体系联合异种神经支架促进神经感觉恢复效果优于运动恢复。

Objective To explore the effect of bone marrow stromal cells(BMSCs) and Schwann cell(SCs) which was co-cultured with acellular nerve xenograft(ANX) onrepairing of sciatic nerve defects in rats.Methods In cell experiments,the SCs were isolated and extracted from left sciatic nerve in SD rats(100±20)g.BMSC sextracted from bilateral femoral and tibial bone in SD rats at four weeks of age. BMSCs and SCs with different concentrations were co-cultured in a Transwell petri dish. The proliferation activity of the cells with different concentrations was detected by CCK-8. Different cell groups were inserted into the preparation scaffold. In animal experiments,rats model with 10 mm sciatic nerve defects was prepared under aseptic conditions,SD rats(220±20 g) were selected for transplantation with compound nerve scaffold into the sciatic nerve defect model,and they were divided into five groups:BMSCs-SCs(1),BMSCs-SCs(2),BMSCs-SCs(3),BMSCs(4) and blank transplantation group. After eight weeks,morphological experiments were performed on each group of rats to evaluate the effect of transplanted nerve regeneration.Results The proliferation activity of BMSCs cells in the co-culture system were stronger than that in the single cell group which showed a typical polar vortex growth. The rats were fed for eight weeks,and then detected the sciatic nerve function index,the hot plate test,the wet weight ratio of the tibial anterior muscle and the gastrocnemius muscle on the affected side/the healthy side of ratswere foundthe co-cultured group had a well recovery of the sciatic nerve function(P<0.05),especially in the sensory area innervated by the sciatic nerve(P<0.05).Conclusion The co-culture system could help the sciatic nerve regeneration and functional recovery;The application of co-culture system combined with acellular nerve xenograft can promote sensory recovery of nerves which was superior to motor recovery.