Department of Chemical Engineering, National Tsing Hua University 101, Section 2, Kuang Fu Road, Hsin Chu, Taiwan 300, Republic of China.
J Biomed Mater Res A. 2010 Feb;92(2):712-23. doi: 10.1002/jbm.a.32198.
This study investigates a poly(epsilon-caprolactone)-graft-type II collagen-graft-chondroitin sulfate (PCL-g-COL-g-CS) biomaterial as a scaffold for cartilage tissue engineering. Biodegradable polyester, PCL, was utilized to fabricate three-dimensional (3D) porous scaffolds by particulate leaching. The PCL scaffold was then surface modified by chemical bonding of 1,6-hexanediamine and the grafting of a bioactive polymer layer of COL and CS with the help of 1-ethyl-3-(3-dimethyl- aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) on the modified PCL surface to produce PCL-g-COL and PCL-g-COL-g-CS, respectively. The characteristics of these modified and grafted matrices were examined by ESCA, aminolysis, collagen and CS assay, porosity and water-binding capacity. Grafted COL and CS markedly increased water-binding capacity, and promoted the spreading and growth of chondrocytes. During a 4-week culture period, PCL-g-COL and PCL-g-COL-g-CS matrices both provided more cell proliferation, as determined by measuring the DNA assay. Additionally, a larger amount of secreted collagen and glycosaminoglycans (GAGs) appeared in the PCL-g-COL-g-CS matrices than in the control (PCL) as indicated by the histochemical sections via Hematoxylin and eosin (H&E) stain, Masson trichrome stain and Safranin-O stain. The chondrocytes were induced to function normally; the cell phenotype was maintained, and the GAGs and collagen in the PCL-g-COL-g-CS scaffold were secreted in vitro. These results serve as a basis for future studies of the fabrication process and reveal the potential biocompatibility of the biomimetic matrix for regenerating articular cartilage or other organs.
本研究探讨了一种聚己内酯接枝型 II 型胶原接枝硫酸软骨素(PCL-g-COL-g-CS)生物材料作为软骨组织工程的支架。可生物降解的聚酯聚己内酯(PCL)通过颗粒沥滤法被用来制造三维(3D)多孔支架。然后,PCL 支架通过 1,6-己二胺的化学键合和 COL 和 CS 的生物活性聚合物层的接枝进行表面改性,在改性的 PCL 表面上使用 1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDC)/N-羟基琥珀酰亚胺(NHS),以分别产生 PCL-g-COL 和 PCL-g-COL-g-CS。这些改性和接枝基质的特性通过 ESCA、氨解、胶原和 CS 测定、孔隙率和水结合能力进行了检测。接枝的 COL 和 CS 显著增加了水结合能力,并促进了软骨细胞的铺展和生长。在 4 周的培养期间,通过测量 DNA 测定,PCL-g-COL 和 PCL-g-COL-g-CS 基质都促进了细胞的增殖。此外,通过苏木精和伊红(H&E)染色、马松三色染色和番红 O 染色的组织化学切片显示,在 PCL-g-COL-g-CS 基质中,比对照(PCL)中出现了更多的分泌型胶原和糖胺聚糖(GAGs)。软骨细胞被诱导正常发挥功能;细胞表型得以维持,并且 PCL-g-COL-g-CS 支架中的 GAGs 和胶原在体外被分泌。这些结果为未来的制造工艺研究提供了基础,并揭示了仿生基质对再生关节软骨或其他器官的潜在生物相容性。