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犬自体骨髓间充质干细胞介导HA_TCP修复牙槽骨缺损的实验研究_牛玉梅

犬自体骨髓间充质干细胞介导HA-TCP修复牙槽骨缺损的实验研究

牛玉梅1* 陈野2 李艳萍1 赵尔杨3 曹涛1 刘会梅1 王爽1 姜凌云1

(1.哈尔滨医科大学口腔医学院牙体牙髓病科 黑龙江哈尔滨 150001;

2.哈尔滨工程大学;3.哈尔滨医科大学口腔医学院口腔病理科)

[摘要] 目的:研究成年犬骨髓间充质干细胞(mesenchymal stem cells,MSCs)介导羟基磷灰石磷酸三钙(hydroxyapatite-tricalcium phosphate,HA-TCP)修复犬牙槽骨缺损。方法:用密度梯度离心贴壁培养法分离纯化成年犬的MSCs,将MSCs矿化诱导为成骨样细胞后与HA-TCP支架复合。选取3只成年犬,在876︱678根分歧处分别制备长宽高均为3mm的骨缺损。左侧作为实验组,牙槽骨缺损处植入MSCs-HA-TCP复合物;右侧作为对照组,将HA-TCP植入牙槽骨缺损部位。移植后8周处死实验动物,进行组织学观察,并进行图像分析。

结果:实验组新生骨组织占缺损的百分比明显高于对照组,剩余材料占缺损的百分比低于对照组,具有统计学意义(P<0.01)。结论:MSCs介导HA-TCP修复成年犬牙槽骨缺损效果优于单纯的HA-TCP修复犬牙槽骨缺损。

[关键词] 骨髓间充质干细胞 牙槽骨缺损 HA-TCP 骨组织工程

[中图分类号] R782.13  [文献标识码] A  [文章编号] 1671—7651(2012)01—0009—03

Repairing of Alveolar Bone Defect with Autoallergic Bone Marrow Mesenchymal Stem Cells Compounded HA-TCP inDogs.NIU Yu-mei,Chen Ye,LI Yan-ping,et al.Department of Operative Dentistry and Endodontics,School of Stomatology,Harbin Medical University,Harbin150001

[Abstract] Objectives:To study the efficacy of repairing the alveolar bone defects with autoallergic bone marrowmesenchymal stem cells(MSCs)compounded HA-TCP(hydroxyapatite-tricalcium phosphate)in dogs.Meth-ods:MSCs of adult dogs were isolated,purified,cultured and induced to osteoblasts.The osteoblasts were implan-ted into the HA-TCP,and then implanted the composite into the defects of mandible in left side of the dogs.TheHA-TCP without MSCs were implanted into right side of the dogs as control.The animals were sacrificed 8weekslater.HE staining and image measurement were performed on implantation section.Results:HE staining and imageanalysis demonstrated that the amount of new alveolar bone formation of experimental group was obviously morethan that of control group.The difference was statistically significant(P<0.01).The residue material of the exper-imental group was fewer than that of the control group.Conclusion:MSCs compounded HA-TCP deployed to re-pair alveolar bone defects in dogs is better than only HA-TCP.

[Key words] MSCs Alveolar bone defect HA-TCP Bone tissue engineering

  因外伤、肿瘤等原因所导致的牙槽骨损伤临床发病率较高,组织工程学的发展为牙槽骨损伤的修复开辟了一条新的途径。本实验应用组织工程学原理,研究骨髓间充质干细胞介导羟基磷灰石磷酸三钙(hydroxyapatite-tricalcium phosphate,HA-TCP)修复犬牙槽骨缺损,旨在为牙槽骨缺损的细胞学治疗及骨髓间充质干细胞构建组织工程化骨提供一定的实验支持。

基金项目 黑龙江省教育厅项目(编
号:11521109)

基金项目 黑龙江省教育厅项目(编号:11521109)

作者简介 牛玉梅(1963~),女,黑龙江人,博士,教授,主要从事牙体牙髓病临床及基础的研究工作。

*通讯作者 牛玉梅,电话:(0451)536455541 材料与方法

1.1 实验材料

1.1.1 主要仪器 MILLI_Q超纯水纯化系统,O-LYMPUS IX70倒置相差显微镜,ECKMAN Al-legr21R恒温离心机,JEOL JEM-1220扫描电子显微镜。

1.1.2 主要材料及试剂 HA-TCP(四川大学材料系提供),LG-DMEM培养液(Hyclone公司),胎牛血清(FBS,匈牙利公司),胰酶(Sigma),地塞米松(Sigma),维生素C(Sigma),β-甘油磷酸钠(Sig-ma),胰岛素(Sigma),3-异丁基-1-甲基黄嘌呤(

 口腔医学研究2012年1月第28卷第1期