Deng Ying, Lin Xing-sun, Zheng Zhong, Deng Jin-Guang, Chen Jin-Chun, Ma Hui, Chen Guo-Qiang
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
Biomaterials. 2003 Oct;24(23):4273-81. doi: 10.1016/s0142-9612(03)00367-3.
The present investigation describes the production of extracellular matrix of rabbit articular cartilage chondrocytes grown on scaffolds of polyhydroxybutyrate (PHB) blended with poly(hydroxybutyrate-co-hydroxyhexanoate) (PHBHHx) for up to 7 days. The mRNA level of type II collagen of chondrocytes seeded on all scaffolds consisting of PHBHHx were obviously higher than that of PHB-only scaffold throughout the culture period, suggesting the positive effect of PHBHHx on extracellular matrix production. Second-harmonic generation (SHG) imaging technique, combined with confocal fluorescence microscopy (CFM) revealed that PHBHHx in PHB scaffold provided better surface properties for anchoring type II collagen filaments and their penetration into internal layers of the scaffolds. Glycosaminoglycan (GAG), a major composition of extracellular matrix, showed a sharp increase in construct of 1:2 PHB/PHBHHx scaffold after 7 day cultivation, while only a small increase was observed in all other tested scaffolds. At the same time, total collagen contents in all scaffolds containing PHBHHx increased with time, with the maximum collagen production of 742.1+/-99.2mg/g dry weight observed in construct of 1:2 PHB/PHBHHx scaffold inoculated for 7 days, this was almost 4-fold higher than that in scaffold of PHB only. It appears that the presence of right proportion of PHBHHx in the composite system of PHB/PHBHHx highly favored the production of extracellular matrix of articular cartilage chondrocytes.
本研究描述了在聚羟基丁酸酯（PHB）与聚（羟基丁酸酯-共-羟基己酸酯）（PHBHHx）混合的支架上培养长达7天的兔关节软骨细胞产生细胞外基质的情况。在整个培养期间，接种在所有由PHBHHx组成的支架上的软骨细胞II型胶原蛋白的mRNA水平明显高于仅PHB支架上的软骨细胞，这表明PHBHHx对细胞外基质产生具有积极作用。二次谐波产生（SHG）成像技术与共聚焦荧光显微镜（CFM）相结合显示，PHB支架中的PHBHHx为II型胶原纤维的锚定及其渗透到支架内层提供了更好的表面特性。糖胺聚糖（GAG）是细胞外基质的主要成分，在1:2 PHB/PHBHHx支架构建体培养7天后显著增加，而在所有其他测试支架中仅观察到少量增加。同时，所有含PHBHHx的支架中的总胶原蛋白含量随时间增加，在接种7天的1:2 PHB/PHBHHx支架构建体中观察到最大胶原蛋白产量为742.1±99.2mg/g干重，这几乎是仅PHB支架的4倍。看来，在PHB/PHBHHx复合体系中存在合适比例的PHBHHx极大地有利于关节软骨细胞细胞外基质的产生。
