纳米羟基磷灰石/聚酰胺66复合骨髓间充质干细胞阻止股骨骨不连的形成  

Nano-hydroxyapatite/polyamide 66 combined with bone marrow mesenchymal stem cells prevents femur nonunion

作　　者：李昂[1] 王晓宇[1] 许茜楠 毕郑刚[1] 

机构地区：[1]哈尔滨医科大学附属第一医院骨科一病房,黑龙江省哈尔滨市150001 [2]哈尔滨医科大学第一临床医学院,黑龙江省哈尔滨市150001

出　　处：《中国组织工程研究》2016年第21期3080-3087,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81171692)~~

摘　　要：背景:骨不连一直是困扰骨科医生的难题,在积极研究如何治疗骨不连的同时更应该注重如何预防它的发生,也就是在治疗高风险发生骨不连的骨创伤时,应该积极干预从而避免发生骨延迟愈合与不愈合。目的:探讨纳米羟基磷灰石/聚酰胺66与大鼠骨髓间充质干细胞复合构建的组织工程骨对股骨骨不连模型病变区域骨愈合的加强与促进作用。方法:(1)体外分离培养大鼠骨髓间充质干细胞分为3组:对照组为未进行骨向诱导的大鼠骨髓间充质干细胞;实验组为经骨向诱导的大鼠骨髓间充质干细胞;复合组将大鼠骨髓间充质干细胞与纳米羟基磷灰石/聚酰胺66复合并进行骨向诱导。通过碱性磷酸酶活性检测以及矿化结节染色验证骨髓间充质干细胞的体外成骨分化能力,扫描电镜观察骨髓间充质干细胞在纳米羟基磷灰石/聚酰胺66上的黏附情况,MTT实验检测细胞的生长与增殖情况;(2)建立SD大鼠股骨骨不连模型后随机分为3组:空白对照组未进行处理,单纯支架组在骨不连区域植入纳米羟基磷灰石/聚酰胺66,细胞复合支架组在骨不连区域植入骨髓间充质干细胞与纳米羟基磷灰石/聚酰胺66复合材料。术后通过X射线、Micro CT、Masson染色评价SD大鼠股骨的骨愈合情况。结果与结论:(1)在成骨诱导第6,12天,实验组碱性磷酸酶活性均显著高于对照组;在成骨诱导第14天,实验组形成的矿化结节数量多于对照组,说明提取的骨髓间充质干细胞体外经骨向诱导后具有成骨分化能力;(2)骨髓间充质干细胞在纳米羟基磷灰石/聚酰胺66上黏附良好,细胞的生长增殖活性良好,说明支架材料纳米羟基磷灰石/聚酰胺66适于骨髓间充质干细胞黏附、增殖并骨向分化;(3)空白对照组大鼠骨不连模型制作成功,12周时骨不连区域几乎无骨痂形成。单纯支架组大鼠在术后8周与12周均未获得骨愈合。细胞复合支架�BACKGROUND： Orthopedists should pay more attentions to nonunion prevention in view of nonunion treatment, that is, active interventions should be taken to avoid bone delayed union and nonunion. OBJECTIVE： To explore the effect of composite tissue-engineered scaffold constructed by nano-hydroxyapatite/polyamide 66（n HA/PA 66） combined with bone marrow mesenchymal stem cells to repair a femoral fracture with severe nonunion. METHODS： Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro, and then they were divided into three groups： bone marrow mesenchymal stem cells without osteogenic induction, with osteogenic induction or combined with n HA/PA 66 followed by osteogenic induction as control group, test group or composite group, respectively. Then osteogenic differentiation of bone marrow mesenchymal stem cells in vitro was analyzed by measuring alkaline phosphatase activity and alizarin red staining, cell adhesion on the n HA/PA 66 was observed using scanning electron microscopy, and the cell growth and proliferation were detected by MTT assay. In the meanwhile, established Sprague-Dawley rat femur nonunion models were randomly divided into three groups： the areas of nonunion were implanted with nothing as blank control group, those were with n HA/PA 66 as simple scaffold group, and the others were with n HA/PA 66 combined with bone marrow mesenchymal stem cells as composite scaffold group. Afterwards, X-ray examination, micro-CT and Masson staining were used to evaluate the femoral healing. RESULTS AND CONCLUSION： At 6 and 12 days after osteogenic induction, alkaline phosphatase activity in the test group was significantly higher than that in the control group; at 14 days, compared with the control group, the amount of mineralized nodules in the test group was significantly higher, which indicated that bone marrow mesenchymal stem cells after osteogenic induction could differentiate into osteoblasts. Attached cells spread well on the scaffold with good proliferation activity,

关 键 词：骨髓 间质干细胞 羟基磷灰石类 生物相容性材料 组织工程 生物材料 骨生物材料 骨髓间充质干细胞 纳米羟基磷灰石/聚酰胺66 生物相容性 骨不连 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]