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Ⅱ型胶原蛋白和硫酸软骨素接枝聚己内酯多孔支架在软骨组织工程中的应用。

The application of type II collagen and chondroitin sulfate grafted PCL porous scaffold in cartilage tissue engineering.

机构信息

Department of Chemical Engineering, National Tsing Hua University 101, Section 2, Kuang Fu Road, Hsin Chu, Taiwan 300, Republic of China.

出版信息

J Biomed Mater Res A. 2010 Feb;92(2):712-23. doi: 10.1002/jbm.a.32198.

Abstract

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.

摘要

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。

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