不同浓度配比的壳聚糖-胶原复合材料对细胞亲和性的影响

Preliminary Study on influence of Chitosan-Collagen Compound with Different ratio for Cell Affinity

目的评价不同浓度配比的壳聚糖-胶原支架材料对细胞亲和性的影响,并观察在细胞-支架复合物培养过程中支架材料的物理性质变化情况。方法采用第3代正常人皮肤成纤维细胞作为种子细胞,进行体外培养和扩增至足够数量后,分别种植于100%胶原(实验组1)、70%胶原(壳聚糖∶胶原=3∶7,实验组2)、50%胶原(壳聚糖∶胶原=5∶5,实验组3)、30%胶原(壳聚糖∶胶原=7∶3,实验组4)四种不同浓度配比的壳聚糖-胶原复合支架材料上,培养7d,通过光镜、电镜等观察和比较细胞在支架上的黏附情况、增殖活力和生长形态,以及材料物理强度的变化情况。结果不同浓度的各组壳聚糖-胶原支架材料上,细胞均能黏附、生长良好,其中实验组2的多孔支架材料上细胞增殖更接近单纯胶原材料(实验组1),且材料收缩降解速度明显低于其他2组。结论不同浓度配比的壳聚糖-胶原支架材料中,壳聚糖∶胶原=3∶7的复合支架材料具有良好的三维空间结构和细胞相容性,且材料形变相对较小。

Objective To investigate the influence of chitosan-collagen compound with different ratio for cell affinity and physical variation of the scaffold. Methods Human dermal fibroblasts of passage 3 were cultured and amplified in vitro, then seeded on the those scaffolds in different mixed ratio groups as follows： 100% collagen as group 1, 70% collagen （chitosan-collagen mixed with 3：7） as group 2, 50% collagen （chitosan-collagen mixed with 5：5） as group 3, 30% collagen （chitosan-collagen mixed with 7：3） as group 4. The attachment efficiency, attachment morphology, proliferation activity and growing morphology of human dermal fibroblasts on all scaffolds were observed by optical microscopy and scanning electronic microscope, and evaluated within 7 days cultivation. Results Cells adhered well on those compound materials in all groups, stretched along the surface of the scaffold and grew when cultured in vitro. More importantly, the cell attachment and proliferation of the group 2 was closed to the pure collagen group 1, which could maintain the shape well with less degradation and contraction. Conclusion The compound scaffold of group 2 with chitosan-collagen mixed with 3：7 has a good cell compatibility and structure. Therefore, this Kind of scaffold may be successfully used for tissue engineering in future.