血小板裂解液联合国产多孔钽对MG63细胞增殖以及ITGβ1/Vinculin/F-actin信号通路表达的影响
PIateIet Iysate combined with domestic porous tantaIum promotes MG63 proIiferation and activates integrin beta1/VincuIin/F-actin signaIing pathway
摘要
Abstract
BACKGROUND: The preliminary study found that domestic porous tantalum is conducive to the early adhesion and proliferation of MG63 cells, which can be used as a scaffold material for bone tissue engineering. As an optimized product of platelet-rich plasma, platelet lysate is more suitable for bone induction in the bone repair. OBJECTIVE: To further investigate the effect of platelet lysate and domestic porous tantalum scaffold constructs on the proliferation of MG63 cells and expression of integrin β1 (ITGβ1)/Vinculin/F-actin signaling pathway based on our previous findings. METHODS: MG3 cells were cultured and inoculated onto domestic porous tantalum scaffolds with the addition of 3%, 5%, 7% and 10% platelet lysates. Then, 7% as the best volume fraction of platelet lysate was screened by cell counting kit-8 method. There were four experimental groups including blank group (normally cultured MG63 cells), platelet lysate group (MG63 cells were cultured in 7% platelet lysate), porous tantalum scaffold group (MG63 cells were cultured on the domestic porous tantalum scaffold), and combined group (MG63 cells were cultured with 7% platelet lysate and porous tantalum scaffold. Scanning electron microscope was used to observe the surface morphology of domestic porous tantalum and platelet lysate-porous tantalum scaffold-MG63 cell complex. Cell counting kit-8 method was used to detect the proliferation of MG63 cells. Real-time fluorescence quantitative PCR (qPCR), immunocytochemical staining and western blot were used to detect the expression of ITGβ1, Vinculin, F-actin in MG63 cells at mRNA and protein levels. RESULTS AND CONCLUSION: Under the scanning electron microscope, MG63 cells adhered well to the scaffold surface. Compared with the blank group, the proliferation of MG63 cells could be significantly promoted by either platelet lysate or porous tantalum scaffold (P < 0.05). Moreover, the proliferation of MG63 cells was significantly improved in the combined group compared with the other three groups (P < 0.05). Findings from qPCR, immunocytochemical staining and western blot showed the highest expression of ITGβ1, Vinculin, F-actin mRNA and protein in the combined group (P < 0.05). These results indicate that platelet lysate and the domestic porous tantalum scaffold can synergistically promote the proliferation of MG63 cells, and up-regulate the expression of ITGβ1, Vinculin and F-actin mRNA and protein. Activation of the ITGβ1/Vinculin/F-actin signaling pathway may contribute to the proliferation, adhesion and differentiation of MG63 cells.关键词
生物材料/骨生物材料/血小板裂解液/多孔钽/MG63细胞/整合素/黏着斑蛋白/束状肌动蛋白分类
医药卫生引用本文复制引用
孙福斋,王少华,邓华民,王茜,李琪佳,甘洪全,王志强..血小板裂解液联合国产多孔钽对MG63细胞增殖以及ITGβ1/Vinculin/F-actin信号通路表达的影响[J].中国组织工程研究,2017,21(34):5430-5436,7.基金项目
国家科技部科技支撑课题(2012BAE06B03) （2012BAE06B03）
河北省科技支撑资助项目(16277776D) （16277776D）
河北省医学科学研究重点课题计划资助项目(20160225) （20160225）
河北省卫生计生委临床医学优秀人才研究项目(361036) （361036）
华北理工大学博士科研启动基金资助项目(286062299) （286062299）
华北理工大学研究生创新项目(2017S40)the National Key Technology Research and Development Program of the Ministry of Science and Technology of China, No. 2012BAE06B03； the Science and Technology Research Project of Hebei Province, No. 16277776D； the Key Medical Research Project of Hebei Province, No. 20160225； the Clinical Talent Research Project of Hebei Provincial Health and Family Planning Commission, No. 361036； the Doctoral Scientific Research Foundation of North China University of Science and Technology, No. 286062299； the Postgraduate Innovation Research Project of North China University of Science and Technology, No. 2017S40 （2017S40）
