Song Yulin, Huang Huabin, Zheng Qixin, Liao Qi
Department of Orthopaedics, Second Affiliated Hospital, Nanchang University, Nanchang 330006, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2009 Dec;26(6):1281-5, 1290.
In this experimental study, the RGD-containing peptide was used to modify the surface of biomimetic PLGA-(ASP-PEG) matrix, and bone marrow stromal cells (BMSCs) were seeded onto these modified surfaces for three weeks. The effects of modified surfaces of matrix on the adhesion, proliferation and differentiation of BMSCs were explored. BMSCs were harvested from whole bone marrow of Sprague-Dawley (SD) rats in vitro, then were seeded onto peptide surface-modified matrix (Experiment group, EG) and matrices without modification (Control group, CG) respectively for three weeks. The number of adhesive cells was counted by using precipitation method after 4 h and 12 h incubation; the cells cytoskeletons were stained with FITC-conjugated phalloidin after 24h incubation; the cell density was investigated after 1 d, 2 d and 3 d of incubation; ALP activity of BMSCs was measured after 7 d, 14 d and 21 d of incubation with osteogenic medium. The cells from bone marrow were BMSCs and their purity was beyond 90% using flow cytometry (FCM) analysis. Sulphur binding energy in EG was shown by XPS to be 164 eV. BMSCs adhered on peptide surface-modified matrix were observed with SEM. Cell adhesion efficiency and quality in EG was better than that in CG, and cell cytoskeleton was more robust in EG. ALP activity was higher in EG than in CG. Peptide surface-modified PLGA-(ASP-PEG) was noted to have good compatibility with BMSCs and to promote cell adhesion and differentiation.
在本实验研究中，使用含RGD的肽修饰仿生PLGA-(ASP-PEG)基质的表面，并将骨髓基质细胞（BMSCs）接种在这些修饰后的表面上培养三周。探讨了基质修饰表面对BMSCs黏附、增殖和分化的影响。体外从Sprague-Dawley（SD）大鼠的全骨髓中获取BMSCs，然后分别接种到肽表面修饰的基质（实验组，EG）和未修饰的基质（对照组，CG）上培养三周。孵育4小时和12小时后，采用沉淀法计数黏附细胞数量；孵育24小时后，用异硫氰酸荧光素（FITC）标记的鬼笔环肽对细胞骨架进行染色；孵育1天、2天和3天后，检测细胞密度；在成骨培养基中孵育7天、14天和21天后，测定BMSCs的碱性磷酸酶（ALP）活性。经流式细胞术（FCM）分析，骨髓来源的细胞为BMSCs，其纯度超过90%。X射线光电子能谱（XPS）显示EG中的硫结合能为164 eV。用扫描电子显微镜（SEM）观察BMSCs在肽表面修饰基质上的黏附情况。EG中的细胞黏附效率和质量优于CG，且EG中的细胞骨架更粗壮。EG中的ALP活性高于CG。结果表明，肽表面修饰的PLGA-(ASP-PEG)与BMSCs具有良好的相容性，能促进细胞黏附和分化。
