胶原-壳聚糖支架与软骨细胞的生长

背景：目前软骨支架材料的种类比较多,但还没有一种材料能完全符合软骨修复的要求。目的：观察在混合材料胶原-壳聚糖支架中软骨细胞的生长情况。方法：采用冷冻干燥法将质量分数为2%胶原与3%壳聚糖混合制备胶原-壳聚糖多孔支架。将分离培养的第2代兔软骨细胞接种到胶原-壳聚糖支架上,对照组将软骨细胞接种到无支架的培养板中。观察支架的孔隙率、吸水性及内部形态结构,MTT法检测软骨细胞在支架上的增殖情况,组织切片苏木精-伊红染色,扫描电镜观察细胞在支架的生长、贴附情况,RT-PCR检测细胞支架复合物蛋白聚糖和Ⅱ型胶原mRNA表达情况。结果与结论：胶原-壳聚糖支架的吸水性为（80.0±0.55）%,孔隙率为（88.5±1.5）%,孔径为100～150μm,复合细胞培养2周后,细胞增殖活力高,软骨细胞分泌的蛋白聚糖和Ⅱ型胶原mRNA表达明显高于对照组。说明质量分数为2%胶原与3%壳聚糖的混合支架适合软骨细胞生长和快速增殖,是一种良好的修复和重建软骨载体。

Chitosan-collagen scaffold and chondrocytes growth

BACKGROUND：At present,the materials of cartilage framework are various,but not one kind of material can fully comply with requirements of cartilage repair.OBJECTIVE：To observe the growth of cartilage cells in mixed chitosan-collagen scaffold.METHODS：The porous chitosan-collagen scaffold was fabricated by the freeze-drying technique using 2% collagen and 3% chitosan.The chondrocytes at passage 2 were planted in chitosan-collagen scaffolds（experimental group） or cell panels（control group）.The porosity,water absorption and internal structure of the scaffold were observed.The cell attachment capacity and proliferation of chondrocytes was valuated by MTT and hematoxylin-eosin staining.Expressions of type Ⅱ collagen and proteoglycan mRNA were valuated by RT-PCR.RESULTS AND CONCLUSION：The water absorption of chitosan-collagen scaffold was（80.0±0.55）%,porosity was（88.5±1.5）%,and the pore size was 100-150 μm.The scaffold facilitated the penetration and attachment of chondrocytes.After two weeks,RT-PCR showed that collagen and proteoglycan mRNA were expressed in the experimental group,but only a little in the control group.These results showed that the chitosan-collagen scaffold（2% collagen and 3% chitosan） can provide an appropriate environment for the generation of chondrocytes and have a potential application in the cartilage tissue engineering scaffold field.It is one kind of good carrier for restoration and reconstruction of cartilage.