不同质量比SF/PLLA复合纤维支架上共培养的上皮细胞和成纤维细胞增殖和表型的变化

Changes of proliferation and phenotypes of epithelial cells and fibroblasts co-cultured on SF/PLLA composite fibrous scaffolds

目的:观察不同质量配比的丝素蛋白(SF)和聚乳酸(PLLA)构建的SF/PLLA复合纤维支架对上皮细胞和成纤维细胞增殖和表型的影响,阐明SF/PLLA生物工程膜的细胞相容性。方法:使用SF和PLLA作为支架材料,以SF与PLLA不同质量比(S50/P50、S40/P60和S30/P70)进行混合纺丝,制备出不同比例的SF/PLLA复合纤维支架,采用SF/PLLA复合纤维支架与皮肤组织中分离的原代上皮细胞和成纤维细胞进行共培养。采用CCK8方法检测细胞在不同质量比的支架材料上的增殖活性,免疫荧光染色观察共培养细胞的特异标记蛋白表达。结果:共培养第5和6天时,在S50/P50组复合纤维支架上的成纤维细胞增殖活性高于S40/P60组和S30/P70组(P<0.01),在S40/P60组复合纤维支架上的上皮细胞增殖活性明显高于S50/P50和S30/P70组(P<0.05或P<0.01)。接种于SF/PLLA复合纤维支架的上皮细胞表达CK19蛋白,接种的成纤维细胞表达Vimentin蛋白。2种细胞与SF/PLLA复合纤维支架共培养,S40/P60和S50/P50组复合纤维支架上的细胞生长状态优于S30/P70组。结论:支架中SF的浓度增加可以明显提高细胞增殖活性,促进细胞存活,SF/PLLA复合纤维支架对细胞的生物表型无明显影响。

Objective To observe the influence of SF/PLLA composite fibrous scaffolds with different quality ratios of SF and PLLA in the proliferation and phenotypes of epithelial cells and fibroblasts,and to clarify the cytocompatibility of SF/PLLA bioengineering membrane.Methods The SF and PLLA were served as materials to construct composite scaffolds.The different proportions of SF/PLLA nanofibers membrane were compounded with different ratios of SF and PLLA（S50/P50,S40/P60 and S30/P70）and mixed spinning.The primary epithelial cells and fibroblasts from rat skin tissue were cultured on the SF/PLLA composite scaffolds.CCK8 test was used to evaluate the cell proliferation,and immunofluorescence staining was used to determine the expressions of the specific marker proteins of cultured cells.Results The proliferation activities of fibroblasts in S50/P50 group were significantly higher than those in S40/P60 and S30/P70 groups,and the proliferation activities of epithelial cells on the composite scaffolds in S40/P60 group were higher than those in S50/P50 and S30/P70groups（P〈0.01）on the5 th and 6th days.The epithelial cells cultured on the SF/PLLA composite scaffolds expressed CK 19 protein and the fibroblasts expressed Vimentin protein. When two kinds of cells were co-cultured on SF/PLLA composite fibrous scaffolds,the cell growth status in S40/P60 and S50/P50 groups were superior to that in S30/P70 group.Conclusion The increasing of SF concentration in composite scaffolds can obviously increase the cell proliferation activity and promote the cell survival;SF/PLLA scaffolds has no significant influence in the cell phenotype.