胶原-壳聚糖支架与软骨细胞的生长

Chitosan-collagen scaffold and chondrocytes growth

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。

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.