重组人骨保护素抑制破骨细胞及促进羟磷灰石修复去势大鼠下颌骨缺损的研究

doi:10.7518/hxkq.2018.04.004

华西口腔医学杂志 ›› 2018, Vol. 36 ›› Issue (4): 367-371.doi: 10.7518/hxkq.2018.04.004

重组人骨保护素抑制破骨细胞及促进羟磷灰石修复去势大鼠下颌骨缺损的研究

廖锋¹(), 刘士博², 刘瑶², 刘航航², 胡建¹, 刘显²()

1.武汉大学口腔医学院,湖北省口腔基础医学重点实验室-省部共建国家重点实验室培育基地,口腔生物医学教育部重点实验室（武汉大学）,武汉 430072
2.口腔疾病研究国家重点实验室国家口腔疾病临床医学研究中心四川大学华西口腔医院口腔颌面外科,成都 610041

收稿日期:2017-10-11 修回日期:2018-03-09 出版日期:2018-08-01 发布日期:2018-08-01
作者简介:
廖锋,主治医师,博士,E-mail：wb003230@whu.edu.cn
基金资助:
国家自然科学基金（31400829）;四川省科技创新团队项目（2017TD0016）

Human osteoprotegerin inhibits osteoclasts and promotes hydroxyapatite to repair the mandibular defects in ovariec-tomized rats

Feng Liao¹(), Shibo Liu², Yao Liu², Hanghang Liu², Jian Hu¹, Xian Liu²()

1. The State Key Laboratory Breeding Base of Basic Science of Stomatology, Hubei Province & Key Laboratory of Oral Biomedicine (Wuhan University), Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan 430072, China
2. State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases & Dept. of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China

Received:2017-10-11 Revised:2018-03-09 Online:2018-08-01 Published:2018-08-01
Supported by:
The National Natural Science Foundation of China (31400829);Sichuan Province Science and Technology Innovation Team Project (2017TD0016)

摘要/Abstract

摘要：

目的探讨转染人骨保护素（hOPG）基因的大鼠骨髓间充质干细胞（rBMSCs）复合羟磷灰石（HA）支架对去势大鼠下颌骨缺损的修复作用。方法将重组腺病毒pDC316-hOPG-EGFP转染rBMSCs,蛋白质印迹法和骨磨片试验分别检测hOPG的表达水平和抑制破骨细胞功能;构建骨质疏松大鼠模型,分别将HA支架、未转染rBMSCs复合HA支架、转染rBMSCs复合HA支架植入大鼠下颌骨骨缺损,6周后通过抗酒石酸酸性磷酸酶、苏木精-伊红染色检测骨缺损区破骨细胞及骨修复情况。结果体外携载有hOPG基因的腺病毒成功转染rBMSCs,转染后的rBMSCs表达具有抑制破骨细胞活性功能的hOPG;表达hOPG的rBMSCs复合HA支架后骨缺损处破骨细胞明显减少,成骨增多。结论转染hOPG基因的rBMSCs在体内外均具有抑制破骨细胞功能的作用,且转染rBMSCs复合HA支架可促进骨质疏松大鼠的下颌骨缺损修复。

关键词: 骨保护素, 骨缺损, 基因转染, 骨再生, 骨质疏松

Abstract:

Objective This study aims to investigate the effect of human osteoprotegerin (hOPG) gene-modified rat bone marrow mesenchymal stem cells (rBMSCs) combined with hydroxyapatite (HA) scaffolds on the repair of mandibular defects in ovariectomized rats. Methods rBMSCs were transfected with adenovirus carrying pDC316-hOPG-EGFP. The expression of hOPG and the inhibition of osteoclast function were detected by Western blot and bone-grinding experiment respectively. The model of mandibular bone defect in rats with osteoporosis was established; HA, untransfected rBMSCs-conjugated HA, and transfected rBMSCs-conjugated HA scaffolds were implanted into the mandibular bone defects. After six weeks, tartrate-resistant acid phosphatase staining and hematoxylin-eosin staining were used to observe the number of osteoclasts and repair of bone defect. Results Adenovirus carrying hOPG gene in vitro were successfully transfected into rBMSCs. The hOPG with anti-osteoclast activity was expressed by hOPG-rBMSCs, and rBMSCs expressing hOPG combined with HA scaffolds promoted mandibular defect repair. Conclusion rBMSCs transfected with hOPG gene inhibited the function of osteoc-lasts both in vitro and in vivo, and transfected rBMSCs com-bined with HA scaffolds promoted the repair of mandibular defects in rats with osteoporosis.

Key words: osteoprotegerin, bone defect, gene transfection, bone regeneration, osteoporosis

中图分类号:

R782

廖锋, 刘士博, 刘瑶, 刘航航, 胡建, 刘显. 重组人骨保护素抑制破骨细胞及促进羟磷灰石修复去势大鼠下颌骨缺损的研究[J]. 华西口腔医学杂志, 2018, 36(4): 367-371.

Feng Liao, Shibo Liu, Yao Liu, Hanghang Liu, Jian Hu, Xian Liu. Human osteoprotegerin inhibits osteoclasts and promotes hydroxyapatite to repair the mandibular defects in ovariec-tomized rats[J]. West China Journal of Stomatology, 2018, 36(4): 367-371.

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参考文献 11

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重组人骨保护素抑制破骨细胞及促进羟磷灰石修复去势大鼠下颌骨缺损的研究

Human osteoprotegerin inhibits osteoclasts and promotes hydroxyapatite to repair the mandibular defects in ovariec-tomized rats

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