BACKGROUND:Studies have shown that bioactive molecules or polypeptides grafted onto the surface of polyethylene glycol (PEG) hydrogels can improve PEG bioactivities.OBJECTIVE:To manufacture PEG hydrogels capable of auto-recruiting growth factor beta1 (TGF-β1) and to study its biocompatibility. METHODS:Pure PEG hydrogels (group A), PEG hydrogels grafted on a cell adhesion peptide RGD peptides (group B), PEG hydrogels grafted with auto-recruited TGF-β1 peptide sensitive polypeptide HSNGLPL (group C), PEG hydrogels grafted with both RGD and HSNGLPL polypeptides (group D) were prepared. Contract angle of the hydrogel was detected in each group. Human bone mesenchymal stem cells were seeded onto four kinds of hydrogels. After cells attached, scanning electron microscope and LIVE/DEAD staining were done to observe cell-hydrogel compounds. Human bone marrow mesenchymal stem cells were co-cultured with ordinary culture medium (control) or four kinds of hydrogels for 1, 3, 5, 7 days, and the cell proliferation was detected by cell counting kit-8 assay. The four kinds of hydrogels were put into 24-well culture plates with addition of PBS containing TGF-β1, and 1 hour later, immunofluorescence staining was done. RESULTS AND CONCLUSION:(1) The contact angles of groups A and C were larger than those of groups B and D. (2) Under the scanning electron microscope, groups A and C had little cells attached on the hydrogel surface, but there were many cells on the hydrogel surface in groups B and D. (3) LIVE/DEAD staining showed groups A and C had little living cells, and conversely groups B and D had many living cells. (4) The results of cell counting kit-8 demonstrated that as the incubation time went on, cell proliferation activity of five different groups increased with no difference at the same time point. (5) Findings from the immunofluorescence staining showed that groups A and B had very weak fluorescence, while groups C and D had stronger green fluorescence. In conclusion, PEG hydrogels grafted with RGD and HSNGLPL polypeptides can auto-recruit TGF-β1, and have good biocompatibility.%背景:有研究将生物活性分子或者多肽接枝到聚乙二醇水凝胶表面,改善聚乙二醇的生物活性.目的:制备可自主结合转化生长因子β1的聚乙二醇水凝胶,观察其生物相容性.方法:分别制备聚乙二醇水凝胶(A组)、接枝细胞黏附多肽RGD的聚乙二醇水凝胶(B组)、接枝HSNGLPL多肽的聚乙二醇水凝胶(C组)、接枝细胞黏附多肽RGD与HSNGLPL多肽的聚乙二醇水凝胶(D组),检测4组水凝胶的接触角.将人骨髓间充质干细胞接种于4组水凝胶中,待细胞贴附后,扫描电镜观察细胞-水凝胶复合物,并进行死活细胞染色.分别采用普通培养基(对照组)、4组水凝胶浸提液培养人骨髓间充质干细胞,培养1,3,5,7 d,采用CCK-8法检测细胞增殖.将4组水凝胶材料放入24孔板中,分别加入含转化生长因子β1的PBS溶液,1 h后进行荧光免疫组织化学染色.结果与结论:①A、C组水凝胶接触角较大,B、D组水凝胶接触角较小;②扫描电镜显示,A、C组水凝胶表面几乎无细胞,B、D组水凝胶表面贴附较多细胞;③死活细胞染色显示,A、C组水凝胶表面几乎无活细胞,B、D组水凝胶表面贴附较多活细胞;④CCK-8法检测结果显示,随着培养时间的延长,5组细胞增殖活性逐渐增强,且5组间细胞增殖无差异;⑤荧光免疫组织化学染色显示,A、B组荧光强度很弱,几乎无荧光;C、D组有较强的绿色荧光;⑥结果表明,接枝细胞黏附多肽RGD与HSNGLPL多肽的聚乙二醇水凝胶,可自主结合转化生长因子β1,具有良好的生物相容性.
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