Ho Ming-Hua, Wang Da-Ming, Hsieh Hsyue-Jen, Liu Hwa-Chang, Hsien Tzu-Yang, Lai Juin-Yih, Hou Lein-Tuan
Department of Chemical Engineering, National Taiwan University, Taipei, Taiwan.
Biomaterials. 2005 Jun;26(16):3197-206. doi: 10.1016/j.biomaterials.2004.08.032.
Chitosan scaffolds were modified with RGDS (Arg-Gly-Asp-Ser) in the present work via an imide-bond forming reaction between amino groups in chitosan and carboxyl groups in peptides. Successful immobilization was verified with FTIR spectroscopy, and the immobilized amount was determined to be on the order of 10(-12) mol/cm2 through analysis of the immobilized amino acids. Results of experiments of cell culture with rat osteosarcoma (ROS) cells demonstrated that RGDS immobilization could enhance the attachment of ROS cells onto the chitosan, resulting in higher cell density attached to the RGDS-modified scaffold than to the unmodified scaffold. It should be noted that only RGDS, but not other peptide such as RGES, is effective in enhancing cell attachment and possible proliferation. Experiments of in vitro mineralization indicated that there were more cells on the RGDS-modified scaffold than on the unmodified scaffold, which tended to form bone-like tissues. The results presented in this work suggest that immobilization of RGDS can make chitosan scaffolds more compatible for the culture of osteoblast-like cells and the regeneration of bone-like tissues.
在本研究中,壳聚糖支架通过壳聚糖中的氨基与肽中的羧基之间形成酰亚胺键的反应,用RGDS(精氨酸-甘氨酸-天冬氨酸-丝氨酸)进行了修饰。通过傅里叶变换红外光谱(FTIR)验证了成功固定,并且通过对固定化氨基酸的分析确定固定化量约为10(-12)mol/cm2。用大鼠骨肉瘤(ROS)细胞进行细胞培养实验的结果表明,RGDS固定化可以增强ROS细胞在壳聚糖上的附着,导致附着在RGDS修饰支架上的细胞密度高于未修饰的支架。应当指出的是,只有RGDS,而不是其他肽如RGES,在增强细胞附着和可能的增殖方面是有效的。体外矿化实验表明,RGDS修饰支架上的细胞比未修饰支架上的细胞更多,这些细胞倾向于形成类骨组织。本研究中呈现的结果表明,RGDS的固定化可以使壳聚糖支架更适合于成骨样细胞的培养和类骨组织的再生。