中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (25): 4611-4614.doi: 10.3969/j.issn.1673-8225.2011.25.013

• 纳米生物材料 nanobiomaterials • 上一篇    下一篇

重组人骨形态发生蛋白2壳聚糖纳米微球的制备及体外细胞毒性

王  玮1,2，尹庆水1，张  余1   

  1. 1解放军广州军区广州总医院骨科医院，广东省广州市  510010
    2南方医科大学研究生院，广东省广州市  510515
  • 收稿日期:2011-03-16 修回日期:2011-05-10 出版日期:2011-06-18 发布日期:2014-01-10
  • 通讯作者: 尹庆水，硕士，教授，主任医师，广州军区广州总医院骨科医院，广东省广州市 510515 gk_yqs@126.com
  • 作者简介:王玮☆，男，1981年生，江西省德兴市人，汉族，2011年南方医科大学毕业，博士，主治医师，主要从事脊柱外科及骨组织工程的研究。 guagualang@sina.com
  • 基金资助:

    全军医学科学技术研究十一五课题(06G045)。

Preparation of recombinant human bone morphogenetic protein 2 loaded chitosan nanosphere and its cytotoxicity in vitro

Wang Wei1,2, Yin Qing-shui1, Zhang Yu1   

  1. 1Orthopedic Hospital of Guangzhou General Hospital of Guangzhou Military Command, Guangzhou  510010, Guangdong Province, China
    2Postgraduate College, Southern Medical University, Guangzhou  510515, Guangdong Province, China
  • Received:2011-03-16 Revised:2011-05-10 Online:2011-06-18 Published:2014-01-10
  • Contact: Yin Qing-shui, Master, Professor, Chief physician, Postgraduate College, Southern Medical University, Guangzhou 510515, Guangdong Province, China gk_yqs@126.com
  • About author:Wang Wei☆, Doctor, Attending physician, Orthopedic Hospital of Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, Guangdong Province, China; Postgraduate College, Southern Medical University, Guangzhou 510515, Guangdong Province, China guagualang@sina.com
  • Supported by:

    Military Medical Research during the Eleventh Five-year Plan, No. 06G045*

PDF

283

被引次数

25

摘要：

背景：通过各种微球负载骨生长因子使骨形态发生蛋白达到缓释效果逐渐成为研究热点，但关于载药壳聚糖纳米微球的生物相容性特别是细胞毒性的报道较少。
目的：对重组人骨形态发生蛋白2壳聚糖纳米微球进行细胞毒性检测，评估应用壳聚糖纳米微球作为重组人骨形态发生蛋白2缓释载体的生物安全性。
方法：通过离子交联法制备空白壳聚糖纳米微球，应用透视电镜观察微球的形态，激光粒径分析其粒径分布；通过重组人骨形态发生蛋白2壳聚糖纳米微球体外细胞毒性试验评估微球的生物安全性。
结果与结论：离子交联法制备的壳聚糖微球，球形规整，分散均匀，微球平均粒径为230 nm，分布较集中。载药及空白微球的反应分级为0或1级，均为合格。提示，离子交联法制备可成功制备出负载重组人骨形态发生蛋2的纳米微球，且微球细胞毒性检测合格，为进一步的骨组织工程研究提供理论实验基础。

关键词: 重组人骨形态发生蛋白2, 壳聚糖, 纳米微球, 制备, 细胞毒性

Abstract:

BACKGROUND: The study on bone morphogenetic protein (BMP) release approach has more important significance, but in recent years studies focus on the release effect of microspheres loaded various bone growth factors. However, the biocompatibility, especially cytotoxicity of drug-loaded chitosan nanoparticles, has not been reported.
OBJECTIVE: To prepare and characterize a controlled release system for recombinant human bone morphogenetic protein 2 (rhBMP-2) and to evaluate the cytotoxicity and biological safety of chitosan nanospheres as a carrier for controlled release of rhBMP-2.
METHODS: The rhBMP-2 loaded chitosan nanospheres were prepared using an ionic crosslinking method with ripolyphosphate  as the crosslinking agent. Transmission electron microscope was used to evaluate the morphological properties and particle size analyzer was used to analysis particle size distribution. A cytotoxic test in vitro was adopted to determine the biological safety of rhBMP-2 loaded chitosan nanospheres.
RESULTS AND CONCLUSION: The rhBMP-2 loaded chitosan nanospheres were spherical in shape, and showed a smooth surface. The mean diameter of nanospheres was 230 nm. The cell relative growth rates (RGR) of rhBMP-2 loaded chitosan nanospheres and blank chitosan nanospheres groups were 0 or 1, and all were qualified. The rhBMP-2 loaded chitosan nanospheres prepared by ionic crosslinking method show a good property and biocompatibility. Such a novel type of controlled release system has the potential of being applied for the bone tissue engineering.

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