生物活性玻璃与壳聚糖复合骨修复材料的制备及细胞相容性的研究  

Preparation and cell compatibility of chitosan composite bioactive glass

作　　者：孙晨[1,2] 祝少博[1] 禹志宏[2] 金林[1] 漆白文[1] 张韬[1] 麦合木提江·穆海麦提 

机构地区：[1]武汉大学中南医院骨科,430071 [2]湖北医药学院附属十堰市人民医院骨科

出　　处：《中华实验外科杂志》2014年第2期368-371,共4页Chinese Journal of Experimental Surgery

基　　金：湖北省教育厅科研计划资助项目（B20122415）;湖北省自然科学基金资助项目（2012FFB02002）

摘　　要：目的 设计一种生物活性玻璃（BG）/壳聚糖（CS）复合材料的骨组织工程支架,检测其理化性能和细胞相容性.方法 2.0％ CS盐酸溶液与β-甘油磷酸钠在冰浴条件下以7∶1比例混合搅拌均匀.电子天平分别称取BG分别加入上述CS溶液中,分别制成理论质量比为2.0∶1.0、1.0∶1.0、1.0∶1.5的CS/BG混合液.冷冻干燥成型,20 kGy60Go辐射灭菌后得到样品.通过扫描电镜、X线衍射和傅立叶红外光谱分析、DSC热分析其微观形貌和组成;计算支架孔孔隙率;采用陶瓷试验系统进行支架的机械性能检测并评价BG的加入对支架的影响.将第3代兔骨髓间充质干细胞接种于支架上,使用扫描电镜检测支架对其黏附作用,采用噻唑蓝（MTT）法检测细胞在支架上增殖,并评价支架的细胞相容性.结果 合成的CS/BG支架与模具拥有同样的大小及几何形状,具有相互贯通的多孔结构,未见BG聚集,孔隙率最高可达89％,孔径大小合适（100 ～ 300μm）,最大断裂强度为（4.90±0.63） mPa.X射线衍射图可以看到特征性的BG衍射峰;傅立叶变换红外光谱可见特征的BG吸收峰,这表明材料内有明确的BG;第3代兔骨髓间充质干细胞在支架上共培养ld后,细胞在支架表面黏附,部分细胞伸展,并伸出伪足.共培养7d后,可见细胞生长良好,沿材料铺展生长,细胞呈多角形,细胞伸出多个细长的伪足与材料相连.采用MTT法测量骨髓间充质干细胞在支架上的增殖,结果表明骨髓间充质干细胞在CS/BG支架上表现出了明显的增殖.结论 采用溶液共混、冷冻干燥法可以制备出CS/BG支架;支架具有良好的孔隙率、较好的机械强度、良好的组织相容性,可用于骨组织工程.Objective To design a kind of bioactive glasses/chitosan （BG/CS） composite materials,and investigate their physico-chemical properties and cell compatibility.Methods 2.0% chitosan of hydrochloric acid solution and β-glycerophosphate on ice conditions in the 7∶1 ratio of well mixed.after CS/BG become 2.0∶ 1.0,1.0∶ 1.0,1.0∶1.5 gelatinous liquid stage.Save it in the-80 ℃ frige,formed by freeze-drying machine.After 20 kGy 60Go radiation sterilization get the sample.By scanning electron microscope,X-ray diffraction and Fourier infrared spectral analysis,DSC thermal analysis,its microstructure and composition,Calculate the stents＇ porosity,adopt ceramic test system of mechanical properties test.Inoculate the third-generation rabbit mesenchymal stem cells into the stents and use electron microscopy scanning the adherency of stents.Detect the proliferation of cells which are adhere to the stents.Evaluate the compatibility of the stents and cells.Results Synthetic CS/BG stents has the same size and geometric shapes with mould.The phenomenon of hydroxyapatite crystals gathered and carbon nanotube reunion was not detected.The porosity is 81%-89%,it is appropriate.The breaking strength is （4.90 ±0.63） mPa,Characteristic BG diffraction peak can be seen through X-ray diffraction figure technology.The absorption peaks of BG can be detected through fourier transform infrared spectroscopy.It indicate that BG is consist in the material.The third generation of rabbit rabbit mesenchymal stem cells adhere to the stent when cultivate for one day,some cells extends and put out locomotions.Training 7 days later,the cells grows well,and spread along the stent,it can be seen that cells polygonal and cell with material connected through slender locomotion.bone marrow mesenchymal stem cell grows well in the CS/BG stents.Conclusion The CS/BG stents can be preparate through the solution blending and freeze drying technology.The stents has good porosity,high mechanical strength and good compatibility with cells.It can be

关 键 词：生物活性玻璃 壳聚糖 组织工程 骨 

分 类 号：R318.08[医药卫生—生物医学工程]