中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (26): 4185-4191.doi: 10.3969/j.issn.2095-4344.2017.26.015

• 药物控释材料 drug delivery materials • 上一篇    下一篇

γ-聚谷氨酸/壳聚糖复合磷酸钙骨水泥制备、表征及性能

疏秀林，施庆珊，陈铭杰，冯  劲
  

  1. 广东省微生物研究所，广东省菌种保藏与应用重点实验室，广东省微生物应用新技术公共实验室和省部共建华南应用微生物国家重点实验室，广东省广州市  510070
  • 收稿日期:2017-04-15 出版日期:2017-09-18 发布日期:2017-09-28
  • 通讯作者: 施庆珊，硕士生导师，广东省微生物研究所，广东省菌种保藏与应用重点实验室，广东省微生物应用新技术公共实验室和省部共建华南应用微生物国家重点实验室，广东省广州市 510070
  • 作者简介:疏秀林，女，1980年生，安徽省铜陵市枞阳县人，汉族，2005年西北农林科技大学毕业，硕士，副研究员，主要从事生物材料方面的研究工作。
  • 基金资助:
    广东省科技计划项目(2013B010102016)；广东省自然科学基金(2014A030313665)

Preparation, characterization and performance of gamma-polyglutamic acid/carboxymethyl chitosan-calcium phosphate cement

Shu Xiu-lin, Shi Qing-shan, Chen Ming-jie, Feng Jin
  

  1. Guangdong Institute of Microbiology, Guangzhou 510070, Guangdong Province, China
  • Received:2017-04-15 Online:2017-09-18 Published:2017-09-28
  • Contact: Shi Qing-shan, Master’s supervisor, Guangdong Institute of Microbiology, Guangzhou 510070, Guangdong Province, China
  • About author:Shu Xiu-lin, Master, Associate investigator, Guangdong Institute of Microbiology, Guangzhou 510070, Guangdong Province, China
  • Supported by:
    the Science and Technology Planning Project of Guangdong Province, No. 2013B010102016, and the Natural Science Foundation of Guangdong Province, No. 2014A030313665

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摘要：

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文题释义：
γ-聚谷氨酸：是一种典型的聚电解质，由D-谷氨酸和L-谷氨酸通过γ-谷氨酰键聚合而成的氨基聚合物，相对分子质量一般在10万-100万。与其他聚合高分子化合物相比，γ-聚谷氨酸在体内能降解为谷氨酸单体，为人体所必需，生物相容性优良，低免疫原性，无毒副作用。虽然γ-聚谷氨酸具有良好的应用前景，但是由于其热稳定性差、加工成型性差、强亲水性(其理化性质易受环境湿度影响)等性质极大地限制了它的应用。
γ-聚谷氨酸/壳聚糖：γ-聚谷氨酸与壳聚糖通过接枝共聚法获得的凝胶体，经冷冻干燥后可得微观多孔复合生物材料。与γ-聚谷氨酸相比，γ-聚谷氨酸/壳聚糖不但具有良好的生物形容性，而且易加工成型，对温度和湿度敏感性低，显著拓宽了γ-聚谷氨酸应用领域。
 
背景：骨缺损修复是一个复杂的过程，将成骨因子施加于骨缺损区域以刺激成骨，已被证明有显著的效果。
目的：制备γ-聚谷氨酸/壳聚糖复合磷酸钙骨水泥，并观察其理化性质，探索其作为生长因子载体的缓释功能。
方法：通过喷雾冷冻干燥法制备γ-聚谷氨酸/壳聚糖凝胶颗粒，装载重组人骨形态发生蛋白2生长因子，与磷酸钙骨水泥的固相通过湿法预混、研磨和冷冻干燥，按比例添加固化液，注模按压成型，即制备复合磷酸钙骨水泥载体。以磷酸钙骨水泥为空白对照，对该复合载体凝固时间、抗压强度、物相分析、微观表征、体外释放性能及生物相容性等进行观察和分析。
结果与结论：①γ-聚谷氨酸/壳聚糖的添加未改变磷酸钙骨水泥的理化性质，各组间初凝时间与终凝时间及抗压强度未见明显差异；②扫描电子显微镜观察改性后的磷酸钙骨水泥呈片状晶体形态，γ-聚谷氨酸/壳聚糖颗粒均匀分散其中；③γ-聚谷氨酸/壳聚糖复合磷酸钙骨水泥能明显促进骨细胞体外增殖；④体外模拟实验表明γ-聚谷氨酸/壳聚糖复合磷酸钙骨水泥具有较好的缓释功能。

关键词: 生物材料, 骨生物材料, γ-聚谷氨酸/壳聚糖, 骨形态发生蛋白2, 磷酸钙骨水泥, 骨组织工程, 广东省自然科学基金

Abstract:

BACKGROUND: Nowadays complex bone defects have become a great challenge to orthopedists. A synergistic contribution of various growth factors and a crosstalk between their signaling pathways have been suggested as determinatives for the overall osteogenic outcome.
OBJECTIVE: To develop calcium phosphate cement (CPC) incorporated with γ-polyglutamic acid/carboxymethyl chitosan (PGA/CMCS), and to evaluate its physical and chemical properties and sustained-release function.
METHODS: The γ-PGA/CMCS polymer composites were prepared by graft copolymerization and spray freeze drying methods, and then loaded with recombinant human bone morphogenetic protein 2 (rhBMP-2) growth factor. CPC served as control group, and γ-PGA/CMCS-CPC containing different contents of rhBMP-2 as experimental groups. A γ-PGA/CMCS-CPC scaffold with regular blade-like crystalline structure was fabricated by injection compression molding. Before mixed with the liquid phase, the solid additives were properly mixed by wet method of CPC solid and the γ-PGA/CMCS carrier, then the pre-blended mix was freeze-dried. The setting time and compressive strength of bone cement in each group were detected, and the microstructure of the material surface was observed under scanning electron microscopy. In vitro release of rhBMP-2 was investigated. The effect of bone cement extracts on cell proliferation was determined through MTS assay.
RESULTS AND CONCLUSION: γ-PGA/CMCS-CPC had the same physicochemical properties to the CPC. Initial and final setting time, compressive strength of bone cement had no significant differences among groups. The scanning electron microscope results showed that the γ-PGA/CMCS-CPC scaffold was covered by regular blade-like crystalline structure and the γ-PGA/CMCS particles were uniformly dispersed in the CPC crystals. A sustained release of rhBMP-2 was observed from the γ-PGA/CMCS-CPC. The cell experiments exhibited that the samples with regular blade-like crystalline structure had better cell response compared to CPC control groups with irregular crystalline structure. These findings indicate that γ-PGA/CMCS-CPC can maintain good physicochemical properties, and release growth factor or drug to promote bone formation.

Key words: Polyglutamic Acid, Chitosan, Calcium Phosphates, Bone Cements, Bone Morphogenetic Proteins, Tissue Engineering

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