Mi Fwu-Long, Shyu Shin-Shing, Peng Chih-Kang, Wu Yu-Bey, Sung Hsing-Wen, Wang Pei-Shan, Huang Chi-Chuan
Department of Applied Science, Chinese Naval Academy, 669 Jiun Shiaw Road, Kaohsiung, Taiwan 813, Republic of China.
J Biomed Mater Res A. 2006 Jan;76(1):1-15. doi: 10.1002/jbm.a.30298.
The development of a novel, three-dimensional, macroporous artificial extracellular matrix (AECM) based on chondroitin sulfate (ChS)-chitosan (Chito) combination is reported. The composite AECM composed of ChS-Chito conjugated network was prepared by a homogenizing interpolyelectrolyte complex/covalent conjugation technique through co-crosslinked with N,N-(3-dimethylaminopropyl)-N'-ethyl carbodiimide (EDC) and N-hydroxysuccinimide (NHS). In contrast to EDC/NHS, two different reagents, calcium ion and glutaraldehyde, were used to react with ChS or Chito for the preparation of ChS-Chito composites containing crosslinked ChS or Chito network in the matrix. The stability and in vitro enzymatic degradability of the glutaraldehyde-, EDC/NHS-, and Ca2+ -crosslinked ChS-Chito composite AECMs were all investigated in this study. The results showed that crosslinking improved the stability of prepared ChS-Chito AECMs in physiological buffer solution (PBS) and provided superior protective effect against the enzymatic hydrolysis of ChS, compared with their non-crosslinked counterpart. Because ChS was a heparin-like glycosaminoglycan (GAG), the ChS-Chito composite AECMs appeared to promote binding efficiency for basic fibroblast growth factor (bFGF). The bFGF releasing from the ChS-Chito composite AECMs retained its biological activity as examined by the in vitro proliferation of human fibroblast, depending on the crosslinking mode for the preparation of these composite AECMs. Histological assay showed that the EDC/NHS-crosslinked ChS-Chito composite AECM, after incorporated with bFGF, was biodegradable and could result in a significantly enhanced vascularization effect and tissue penetration. These results suggest that the ChS-Chito composite AECMs fabricated in this study may be a promising approach for tissue-engineering application.
报道了一种基于硫酸软骨素（ChS）-壳聚糖（Chito）组合的新型三维大孔人工细胞外基质（AECM）的开发。由ChS-Chito共轭网络组成的复合AECM通过均质化聚电解质复合物/共价共轭技术，与N,N-（3-二甲基氨基丙基）-N'-乙基碳二亚胺（EDC）和N-羟基琥珀酰亚胺（NHS）共交联制备而成。与EDC/NHS不同，使用两种不同的试剂钙离子和戊二醛与ChS或Chito反应，以制备在基质中含有交联ChS或Chito网络的ChS-Chito复合材料。本研究对戊二醛、EDC/NHS和Ca2+交联的ChS-Chito复合AECM的稳定性和体外酶降解性进行了研究。结果表明，交联提高了制备的ChS-Chito AECM在生理缓冲溶液（PBS）中的稳定性，与未交联对应物相比，对ChS的酶水解提供了更好的保护作用。由于ChS是一种类肝素糖胺聚糖（GAG），ChS-Chito复合AECM似乎提高了碱性成纤维细胞生长因子（bFGF）的结合效率。从ChS-Chito复合AECM释放的bFGF通过人成纤维细胞的体外增殖检测，保留了其生物活性，这取决于制备这些复合AECM的交联模式。组织学分析表明，EDC/NHS交联的ChS-Chito复合AECM在掺入bFGF后可生物降解，并可导致显著增强的血管化效应和组织穿透。这些结果表明，本研究制备的ChS-Chito复合AECM可能是一种有前途的组织工程应用方法。
