骨髓基质干细胞与功能化自组装多肽的生物相容性研究

Biocompatibility of functionalized self-assembling peptide hydrogel with bone mesenchymal stem cells in vitro

目的检测骨髓基质干细胞(BMSCs)在功能化自组装多肽水凝胶中分化及生物相容性。方法自组装多肽RADA16与多肽RADA16-IKVAV委托上海吉尔生物科技公司合成,将两者混合制备功能化自组装多肽水凝胶,用原子力显微镜(AFM)观察其形态学特征。体外分离、扩增、纯化鼠BMSCs并流式分析其表面抗原标志,将BMSCs分别接种到接种到RADA16自组装多肽水凝胶(对照组)与功能化自组装多肽水凝胶(实验组)表面。激光扫描共聚焦显微镜(LSCM)观察细胞迁移情况。应用Nestin、MAP2、GFAP免疫荧光染色,检测骨髓基质干细胞分化情况。结果 AFM显示功能化自组装多肽水凝胶由纳米纤维组成,其直径为7.0~8.0 nm。分离的BMSCs细胞表面标志CD90、CD44、CD34和CD45的阳性率分别为93.71%、92.5%、1.1%、2.18%。共培养后实验组中BMSCs的增殖及迁移能力均明显高于对照组(<0.05);实验组中MAP2阳性细胞百分率较对照组显著提高(<0.05),GFAP阳性细胞百分率较对照组显著降低(<0.05)。结论功能化自组装多肽水凝胶具有良好的细胞生物相容性,从而为神经组织工程提供了一个有前景的生物材料。

Objective To Investigate the potential and differentiation of bone marrow stromal cells which in functional self-assembling peptide hydrogel. Methods The functionalized self-assembling peptide was made by mixing the self-assembling peptide RADA16 and peptide RADA16-IKVAV which was synthesized by the Shanghai Jier Biological Technology Company. The morphological features of the functionalized self-assembling peptide were studied by atom force microscope(AFM). In vitro isolation, amplification, purification rat BMSCs, And separated and identified by flow cytometry for the cell surface markers.BMSCs were seeded on the surface of both the functionalized self-assembling peptide hydrogel(experimental group) and RADA16 self-assembling peptide hydrogel(control group). The cell migration was detected by laser scan confocal microscope(LSCM). Immunofluorescence staining with Nestin, MAP2, and GFAPwas used to assess the differentiation of BMSCs.Results AFM showed that the functionalized self-assembling peptide hydrogel was made up of nanofibers with diameter of 7.0 to 8.0 nm.The surface markers of CD90, CD44, CD34 and CD45 on MSCs were 93.71%, 92.5%, 1.1% and 2.18% respectively.The power of cell proliferation and migration in experimental group was significantly higher than that of control group( <0.05). Compared with control group, the percentage of MAP2 positive cells increased significantly( <0.05) and the percentage of GFAP positive cells decreased significantly( <0.05) in experimental group. Conclusion The functionalized self-assembling peptide hydrogel has good biocompatibility with NSCs, suggesting that it may be a promising biomaterial for neural tissue engineering.