中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (25): 4583-4586.doi: 10.3969/j.issn.1673-8225.2011.25.007

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

缓释型重组人骨形态发生蛋白2/壳聚糖生物骨修复材料诱导骨形成

王丁丁1，曾  戎2，杨敏儿1，何鉴贤1，陈  阳3，何水连3，陈安安3，周智优3，袁丽颖4，汪  炬3   

  1. 1广东药学院生命科学与生物制药学院，广东省广州市 510006；暨南大学，2材料科学与工程系，3生命科学技术学院，广东省广州市 510632；4吉林大学附属第三幼儿园，吉林省长春市 130021
  • 收稿日期:2011-03-04 修回日期:2011-03-29 出版日期:2011-06-18 发布日期:2014-01-10
  • 通讯作者: 汪炬，博士，副研究员，暨南大学生命科学技术学院，广东省广州市510632 wang_ju1688@yahoo.cn
  • 作者简介:王丁丁☆，女，1979年生，贵州省遵义县人，汉族，2007年吉林大学毕业，博士，讲师，主要从事基因工程药物研究。 wangdingding305@163.com
  • 基金资助:

    广东省科技攻关项目(2009B030801237)；广东省教育部科技部企业科技特派员行动计划项目(2009B090600134)；国家自然科学基金青年基金项目(81000923)。

Enhancement of bone formation by recombinant human bone morphogenetic protein-2/chitosan bone biomaterials

Wang Ding-ding1, Zeng Rong2, Yang Min-er1, He Jian-xian1, Chen Yang3, He Shui-lian3, Chen An-an3, Zhou Zhi-you3, Yuan Li-ying4, Wang Ju3   

  1. 1Institute of Life Science and Biological Pharmacy, Guangdong Pharmaceutical University, Guangzhou  510006, Guangdong Province, China
    2Department of Materials Science and Engineering, Jinan University, Guangzhou  510632, Guangdong Province, China
    3College of Life Science and Technology, Jinan University, Guangzhou  510632, Guangdong Province, China
    4The Third Kindergarten Affiliated to Jilin University, Changchun  130021, Jilin Province, China
  • Received:2011-03-04 Revised:2011-03-29 Online:2011-06-18 Published:2014-01-10
  • Contact: Wang Ju, Doctor, Associate investigator, College of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong Province, China wang_ju1688@yahoo.cn
  • About author:Wang Ding-ding☆, Doctor, Lecturer, Institute of Life Science and Biological Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, Guangdong Province, China wangdingding305@163.com
  • Supported by:

    Tackle Key Program of Guangdong Province, No. 2009B030801237*; Action Plan of Enterprise Technology Correspondent, Guangdong Educational Bureau & Guangdong Science and Technology Bureau, No. 2009B090600134*; the National Natural Science Foundation of China, No. 81000923*

PDF

412

被引次数

25

摘要：

背景：重组人骨形态发生蛋白2在体内半衰期短、易降解代谢，达不到理想的骨再生效果。
目的：制备缓释型重组人骨形态发生蛋白2/壳聚糖生物骨修复材料，并观察其缓释性能、骨诱导活性。
方法：将重组人骨形态发生蛋白2与壳聚糖混合制备壳聚糖膜，涂覆于生物骨修复材料表面，ELISA方法检测其体外释药性能。茜素红染色检测缓释型人骨形态发生蛋白2/壳聚糖生物骨材料、重组人骨形态发生蛋白2生物骨材料、单纯骨填充材料诱导C2C12细胞骨钙蛋白的形成，观察其诱导成骨细胞能力。同时将3种骨修复材料植入清洁级KM小鼠股部肌袋内，2周后检测新生骨Ca2+离子含量，评价其异位骨诱导能力。
结果与结论：材料表面的壳聚糖膜分布均匀，负载的重组人骨形态发生蛋白2呈团簇状。重组人骨形态发生蛋白2/壳聚糖生物骨修复材料体外释药存在突释，前4 d释放量达总药量的50%，持续至12 d，释药量达到90%，第18天时释放完全。与单纯骨填充材料、重组人骨形态发生蛋白2生物骨材料相比，缓释型人骨形态发生蛋白2/壳聚糖生物骨修复材料诱导C2C12细胞向成骨晚期分化能力与异位骨形成能力显著增强(P < 0.05)。结果提示缓释型人骨形态发生蛋白2/壳聚糖生物骨修复材料缓释性能好，促进骨形成能力强。

关键词: 人骨形态发生蛋白2, 壳聚糖, 缓释, 骨修复材料, 骨形成

Abstract:

BACKGROUND: The recombinant human bone morphogenetic protein-2 (rhBMP-2) administered in solution often loses its bioactivity in a short time and does not always exhibit efficacy required in bone regeneration in vivo. It would be ideal to develop a system for the sustained delivery of biologically active rhBMP-2 over an extended period of time.
OBJECTIVE: To evaluate rhBMP-2 sustained delivery and osteoinductive effects of bone biomaterials which were combined with chitosan films containing rhBMP-2.
METHODS: ①The rhBMP-2 was added into a chitosan solution to prepare chitosan films which were spread on the bone materials. The effect of slow release was analyzed by ELISA assays. ②In vitro, the osteoinductive to differentiation marker (osteocalcin) of C2C12 cells after induction of rhBMP-2/chitosan (CS) bone biomaterials was assayed by Alizarin red staining.  ③Intramuscular implantation test was made in vivo, rhBMP-2/CS carriers were implanted in muscles of the hind leg of rat for two weeks. The amount of calcium deposited in the implants was measured to assay bioactivity.
RESULTS AND CONCLUSION: ①A scanning electron micrograph of the rhBMP-2/CS bone biomaterials showed uniform distribution of chitosan films on the material surface with rhBMP-2 scattering like cluster. ②An abrupt rhBMP-2 release from rhBMP-2/CS carriers was observed. 50% of the loaded rhBMP-2 was released in 4 days. Thereafter, the release rate was nearly constant until day 12 when 90% of rhBMP-2 was released. All of it was released by day 18 at last. ③In vitro rhBMP-2 released from the rhBMP-2/CS carriers stimulated an increase in osteocalcin of C2C12 cells after 4 weeks. ④In vivo bone formation studies showed the rhBMP-2/CS biomaterials induced bone formation to a much greater extent than control groups by Ca2+ test assay. These results demonstrate that the rhBMP-2/CS bone biomaterials delivery system is capable of potentiating the osteogenic efficacy of rhBMP-2 and underscore its importance as a possible bone regeneration strategy.

中图分类号: