不同孔径聚乳酸-壳聚糖-明胶支架对成纤维细胞粘附效果的研究

Research on the Adhesion Effect of L929 in the Different Pore Size PLA-Chitosan-Gelatine Scaffolds

目的:比较L929细胞在聚乳酸-壳聚糖-明胶(PLA-Chitosan-Gelatin)梯度孔径、均匀孔径两种支架材料上的细胞接种率,选择细胞接种率较高的支架材料;通过比较不同细胞密度在聚乳酸-壳聚糖-明胶梯度孔径支架上的细胞粘附量,探究细胞接种的适宜密度。方法:在两种孔径支架材料上分别接种不同密度的细胞悬浮液,MTT法检测其粘附情况;调整细胞接种密度到同一梯度孔径支架材料上,MTT法比较不同密度细胞在同一梯度孔径支架上的粘附情况。结果:L929细胞在两种支架上均能粘附,在聚乳酸-壳聚糖-明胶梯度孔径支架上的接种率明显高于均匀孔径支架(P<0.01),且随接种密度的增高,粘附到梯度孔径支架上的细胞数量增加,当细胞接种密度达1.0×106/mL时,支架上粘附的细胞数量最大。结论:聚乳酸-壳聚糖-明胶梯度孔径较均匀空径支架材料细胞接种率高,1.0×106/mL为L929细胞在梯度孔径支架上的较适接种密度。

Objective： To select a more appropriate structure of scaffold for cell attach to select appropriate concen- tration for inoculation of L929 cells on PLA-Chitosan-Gelatin 3-D scaffold with pore size gradient, and to obtain the largest amount of initial cell on the scaffolds. Methods： We seeded the cells with different concentration on the scaffolds with pore size gradient and uniform pore size respectively, and cultivate it in coincubator box about 4 hours and count the amount of the cells attached on the scaffold by MTT method. Results： L929 can attach on the scaf- folds. The seeding efficiency is higher on the scaffold with pore size gradient than pore size uniformity （P〈0.01）. The number of the cells attached on the scaffold is the largest when the cell seeding concentration is 1.0×10^6/ml. Conclusion： PLA-Chitosan-Gelatin 3-D scaffold with pore size gradient can raise the seeding efficiency. It is po- tential scaffold for gingival tissue engineering. 1.0 ×10^6/ml is the most suitable cell seeding concentration.