Li Zhensheng, Zhang Miqin
Department of Materials Science and Engineering, University of Washington, 302L Roberts Hall, Seattle, Washington 98195, USA.
J Biomed Mater Res A. 2005 Nov 1;75(2):485-93. doi: 10.1002/jbm.a.30449.
Tissue compatibility of chitosan-alginate scaffolds was studied in vitro in terms of cell morphology, proliferation, and functionality using HTB-94 cells. The scaffold has an interconnected 3D porous structure, and was fabricated by thermally induced phase separation followed by freeze drying. Cell proliferation on the chitosan-alginate scaffold was found to be faster than on a pure chitosan scaffold. After cell culture for 2 weeks in vitro, the cells on the chitosan scaffold gradually assumed a fibroblast-like morphology while the cells on the chitosan-alginate scaffold retained their spherical morphology throughout the period of study. SDS-PAGE electrophoresis and Western blot assays for proteins extracted from cells grown on scaffolds indicated that production of cartilage-specific collagen type II, a marker for chondrocytic phenotype, increased from week 2 to week 3 on the chitosan-alginate scaffold but decreased on the chitosan scaffold. This study suggested that chitosan-alginate scaffolds promote cell proliferation, enhance phenotype expression of HTB-94 chondrocytes, and may potentially serve as an improved alternative to chitosan scaffolds for cartilage tissue engineering.
使用HTB - 94细胞，从细胞形态、增殖和功能方面在体外研究了壳聚糖 - 海藻酸盐支架的组织相容性。该支架具有相互连接的三维多孔结构，通过热诱导相分离然后冷冻干燥制备而成。发现壳聚糖 - 海藻酸盐支架上的细胞增殖比纯壳聚糖支架上的更快。在体外细胞培养2周后，壳聚糖支架上的细胞逐渐呈现成纤维细胞样形态，而壳聚糖 - 海藻酸盐支架上的细胞在整个研究期间保持其球形形态。对在支架上生长的细胞提取的蛋白质进行SDS - PAGE电泳和蛋白质印迹分析表明，软骨特异性II型胶原蛋白（软骨细胞表型的标志物）的产生在壳聚糖 - 海藻酸盐支架上从第2周增加到第3周，但在壳聚糖支架上减少。这项研究表明，壳聚糖 - 海藻酸盐支架促进细胞增殖，增强HTB - 94软骨细胞的表型表达，并且可能作为软骨组织工程中壳聚糖支架的一种改进替代品。
