中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (21): 3080-3087.doi: 10.3969/j.issn.2095-4344.2016.21.006

• 组织工程骨及软骨材料 tissue-engineered bone and cartilage materials • 上一篇    下一篇

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

李 昂1，王晓宇1，许茜楠2，毕郑刚1   

  1. 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
  • 收稿日期:2016-03-26 出版日期:2016-05-20 发布日期:2016-05-20
  • 通讯作者: 毕郑刚，教授，博士生导师，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
  • 作者简介:李昂，男，1989年生，黑龙江省哈尔滨市人，汉族，硕士，主要从事骨组织工程方面的研究。
  • 基金资助:

    国家自然科学基金项目(81171692)

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

Li Ang1, Wang Xiao-yu1, Xu Qian-nan2, Bi Zheng-gang1   

  1. 1First Ward Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China; 2First School of Clinical Medicine, Harbin Medical University, Harbin 150001, Heilongjiang Province, China
  • Received:2016-03-26 Online:2016-05-20 Published:2016-05-20
  • Contact: Bi Zheng-gang, Professor, Doctoral supervisor, First Ward Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China
  • About author:Li Ang, Master, First Ward Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81171692

PDF

210

被引次数

4

摘要：

 文章快速阅读：

文题释义：

 

纳米羟基磷灰石/聚酰胺66：将羟基磷灰石与聚酰胺66进行共融而制成的支架材料，其中羟基磷灰石为无机成分，与人骨的无机成分相同，聚酰胺66是有机成分，与人骨的主要有机成分胶原分子结构相似，因此该材料充分模拟人骨。纳米羟基磷灰石/聚酰胺66具有良好的生物相容性、骨传导性、力学特性并且具备适宜的孔径与孔隙率，在骨组织工程中具有良好的应用前景。

 

骨不连：美国食品与药品管理局(Food and Drug Administration，FDA)将损伤和骨折后至少9个月不愈合，并且没有进一步愈合倾向已有3个月，称为骨不连。骨不连无法通过继续延长治疗时间而获得愈合，需要再次手术治疗，给患者带来痛苦与经济负担。

 

 

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。细胞复合支架组大鼠术后8 周未获得骨愈合，但是有大量骨痂形成，术后12周基本获得骨愈合，支架材料被骨组织包埋与吸收，说明骨髓间充质干细胞与纳米羟基磷灰石/聚酰胺66复合构建的组织工程骨可以促进大鼠股骨骨不连模型中骨折的愈合，有效地阻止了骨不连的发生。

 

 

 

关键词: 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

Abstract:

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 (nHA/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 nHA/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 nHA/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 nHA/PA 66 as simple scaffold group, and the others were with nHA/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, suggesting that nHA/PA 66 is suitable for cell adherence, proliferation and osteogenic differentiation. Besides, at 12 weeks after modeling, in the blank control group, no callus appeared in the nonunion region. In the simple scaffold group, the broken femur did not heal at 8 and 12 weeks after surgery. In the composite scaffold group, the broken femur did not heal at 8 weeks, but a lot of callus appeared; at 12 weeks, bone healing achieved and the scaffold was encased and absorbed. These findings demonstrate that the tissue-engineered bone scaffolds constructed by bone marrow mesenchymal stem cells and nHA/PA 66 effectively prevent bone nonunion by accelerating femoral healing in a rat femur nonunion model.

 

Key words: Bone Marrow, Mesenchymal Stem Cells, Hydroxyapatites, Biocompatible Materials, Tissue Engineering

中图分类号: