宋克东,刘天庆,郭文华,梅冠宇,姜丽丽,马学虎,崔占峰

• 1:大连理工大学化工学院干细胞与组织工程研发中心
• 2:牛津大学工程科学系组织工程与生物处理中心

摘要(Abstract)：

为解决组织工程支架材料内部营养供应不足的问题,将营养物质包埋在聚合物微球内再植入支架,通过微球内营养物质的控制释放保持支架内部营养物质浓度的持续均匀性。以牛血清白蛋白(BSA)为模型蛋白,乳酸-羟基乙酸共聚物(PLGA)为外包被材料,采用复乳法(w/o/w)制备聚合物微球,然后将该微球与壳聚糖溶液混合,冻干形成支架。以BCA法检测载药微球自身的释放情况以及其植入到支架后的释放情况,通过扫描电镜观察载药微球的结构以及其植入支架材料后结构变化。结果表明,微球形态圆整,粒径范围在27～55μm之间。壳聚糖支架呈多孔状结构,1%壳聚糖支架孔隙率、吸水率和降解率分别为(92.99±2.51)%、(89.66±0.66)%和(73.77±3.21)%。体外释放显示:微球可以持续释放所包埋的蛋白,突释较小,168h后PLGA微球的累积释放量为(20.24±0.83)%。电镜照片显示,微球植入支架后,与支架结合紧密,微球结构没有发生明显改变。168h后支架内部蛋白浓度为(11.44±1.81)×10-2mg·mL-1。相比传统靠外部培养基自由扩散到支架内部为支架内细胞供养的方法,营养物质或细胞因子包埋在控释微球中并与支架材料复合成可控释支架,可以长时间维持支架中这些因子的浓度均匀,从而为组织工程提供理想的支架材料。

关键词(KeyWords)： 壳聚糖;支架材料;PLGA;微球;BSA;控制释放

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(30670525,30700181);; 教育部博士点新教师基金(20070141055);; 大连理工大学青年教师培养基金(893228)资助

作者(Author): 宋克东,刘天庆,郭文华,梅冠宇,姜丽丽,马学虎,崔占峰

参考文献(References)：

• [1]Tabata Y.The importance of drug delivery systems in tissue engineering[J].Pharm Sci Technol Today,2000,3(3):80-89.
• [2]CHEN Ying-jie(陈英杰),XU Xiang-yong(许向阳),ZHOU Jian-ping(周建平),et al.Advances in reasearching on poly(lactice-co-glycolic acid)microspheres(乳酸-羟基乙酸共聚物微球的研究进展)[J].Journal of China Pharmaceutical University(中国药科大学学报),2007,38(2):186-189.
• [3]Liu J,Meisner D,Kwong E,et al.A novel trans-lymphatic drug delivery system:Implantable gelatin sponge impregnated with PLGA-paclitaxel microspheres[J].Biomaterials,2007,28(21):3236-3244.
• [4]Jaklenec A,Wan E,Murray M E,et al.Novel scaffolds fabricated from protein-loaded microspheres for tissue engineering[J].Biomaterials,2008,29(2):185-192.
• [5]Fu K,Harrell R,Zinski K,et al.A potential approach for decreasing the burst effect of protein from PLGA microspheres[J].J Pharm Sci,2003,92(8):1582-1591.
• [6]Ana Jaklenec,Alexandra Hinckfuss,Bahar Bilgen,et al.Sequential release of bioactive IGF-I and TGF-β1from PLGA microsphere-based scaffolds[J].Biomaterials,2008,29(10):1518-1525.
• [7]HANG Gang(韩钢),CHEN Hai-liang(陈海靓),SUN Xiao-yi(孙晓译),et al.Preparation and release behavior of chitosan scaffolds encapsulating proteins loaded in PLGA microspheres(包埋PLGA微球壳聚糖支架的构建及其对蛋白释放的调节)[J].Acta Pharmaceutica Sinica(药学学报),2006,41(6):493-497.
• [8]JIANG Li-qin(姜丽琴),ZHANG Guo-liang(张国亮),ZHANG Feng-bao(张凤宝),et al.Chitosan membranes used for bilirubin removal(用于清除胆红素的壳聚糖膜的性能研究)[J].J Chem Eng of Chinese Univ(高校化学工程学报),2003,17(2):128-133.
• [9]Eugene K,Lee Y L.Implantable applications of chitin and chitosan[J].Biomaterials,2003,24(13):2339-2349.
• [10]Gravel M,Gross T,Vago R,et al.Response of mesenchymal stem cell to chitosan-coralline composites microstructured using coralline as gas forming agent[J].Biomaterials,2006,27(9):1899-1906.
• [11]Yang Y Y,Chia H H,Chung T S.Effect of preparation temperature on the characteristics and release profiles of PLGA microspheres containing protein fabricated by double emulsion solvent extraction/evaporation method[J].J Control Release,2000,69(1):81-96.
• [12]Pean J M,Venier-Julienne M C,Boury F,et al.NGF release from poly(D,L-lactide-co-glycolide)microspheres.Effect of some formulation parameters on encapsulated NGF stability[J].J Control Release,1998,56(123):175-187.
• [13]WEN Peng-fei(文鹏飞),SONG Keng-dong(宋克东),LIU Tian-qing(刘天庆),et al.Cell adhesion behavior,fabrication and surface-modification of chitosan scaffolds(壳聚糖支架材料的制备、表面修饰及细胞粘附性能的研究)[J].J Chem Eng of Chinese Univ(高校化学工程学报),2008,22(6):992-998.
• [14]Wang Y C,Lin M C,Wang D M,et al.Fabrication of a novel porous PGA-chitosan hydrid matrix for tissue engineering[J].Biomaterials,2003,24(6):1047-1057.
• [15]Song E,Kim S Y,Chun T H,et al.Collagen scaffolds derived from a marine source and their biocompatibility[J].Biomaterials,2006,27(15):2951-2961.
• [16]GU Hai-gang(谷海刚),JIN Xu(金旭),LONG Da-hong(龙大宏),et al.Effect of formalution parameters and additives on BSA encapsulation yields in PLGA microspheres(BSA-PLGA微球的制备条件优化及不同添加剂对包封率的影响)[J].Chinese Journal of Biomedical Engineering(中国生物医学工程学报),2007,26(6):931-935.
• [17]ZHENG Cai-hong(郑彩虹).Design of Biodegradable Microspheres for Protein(蛋白类生物可降解微球的设计)[D].Hangzhou(杭州):Zhejiang University(浙江大学),2006.