雪旺细胞对骨组织工程支架材料复合成骨细胞修复兔桡骨缺损的影响

Effect of Schwann Cells on Repairing Rabbit Radius Defect with Tissue Engineered Bone Combined with Osteoblasts

【目的】探讨雪旺细胞对骨组织工程支架材料复合成骨细胞修复兔桡骨缺损的影响。【方法】分离培养免骨髓间充质干细胞（BMSCs），诱导分化为成骨细胞（IOBs），并从乳兔中提取雪旺细胞（SCs），S-100免疫荧光染色鉴定。采用新西兰大白兔建立桡骨缺损模型，随机平均分成5组：支架材料组（B组）在骨缺损处植入纳米壳聚糖／羟基磷灰石多孔支架；IO＆／支架材料组（C组）在骨缺损处植入种植10＆的支架材料；SCs＋支架材料组（D组）骨缺损处植入种植SCs的支架材料，IOBs／SCs／支架材料组（E组）：骨缺损处植入同时种植IOBs和SCs的支架材料；空白对照组（A组）在骨缺损处不做任何处理。各组在术后8周评估各组大白兔桡骨缺损部位新生骨的形成情况。【结果】术后8周x射线检测结果显示：B组、C纽、D组骨缺损部分愈合，C组、D组优于B组，E组骨缺损完全愈合，而A组骨缺损未愈合。HE染色结果显示：8周时同B组或c组或D组比较，E组成骨细胞和骨小梁数量最多，骨修复最快；免疫组化检测结果显示：脑源性神经生长因子（BI）NF）和神经生长因子（NGF）在E组中的表达较其他组别明显增多。【结论]SCs能够显著促进工程支架材料复合成骨细胞修复兔桡骨缺损，同神经营养因子BDNF和NGF表达紧密相关。

[Objective]To observe the effect of Schwann cells on repairing radius defects with bone tissue engineering scaffolds composited with osteoblasts in rabbit. [Methods]Bone marrow mesenchymal stem cells （13MSCs） were isolated and cultured, then induced to differentiate into osteoblasts （IO13s）. Schwann cells （SCs） were extracted from neonatal rabbit, which were identified by S-100 immunofluorescence. New Zealand rabbits were used to establish the model of ra- dial defect. The models were randomly divided into 5 groups, group A was blank control group, while bone defects were implanted with nano-chitosan/hydroxyapatite porous scaffolds in group 13. and bone defects were implanted with osteo- blasts and scaffolds in group C. Furthermore, bone defects were implanted with Schwann cells and scaffolds in group D. and bone defects were implanted with Schwann cells, osteoblasts and scaffolds in group E. After 8 weeks, immunohisto- chemistry, HE staining, gross morphology and X-ray were performed to evaluate the formation of new bone in the radial defect of rabbits in each group. [Results] The results of X-ray examination after 8 weeks postoperatively showed that the bone defect was partially healed in ]3, C and D groups. Groups C and D showed better effects than group B. The bone de- feet in group E was healed completely, while the bone defect in group A wererft healed at all. Compared to group 13 or C or D after 8 weeks, HE staining showed that the number of osteoblasts and trabecular meshwork were the highest in group E. Also, bone repair was the fastest in group E. lmmunohistochemistry showed that the expression of 13DNF and NGF in group E were signifieantly higher than other groups. [Conclusion]Schwarm ceils （SCs） can significantly promote the repair of rabbit radius bone defects by engineering scaffold composited with osteoblasts, which was closely related to the expression of 13DNF and NGF.