Chinese Journal of Tissue Engineering Research ›› 2017, Vol. 21 ›› Issue (34): 5430-5436.doi: 10.3969/j.issn.2095-4344.2017.34.004

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Platelet lysate combined with domestic porous tantalum promotes MG63 proliferation and activates integrin beta1/Vinculin/F-actin signaling pathway

Sun Fu-zhai1, 2, Wang Shao-hua1, 2, Deng Hua-min1, Wang Qian3, Li Qi-jia3, Gan Hong-quan1, Wang Zhi-qiang1
  

  1. 1Department of Orthopaedics, Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, Hebei Province, China; 2Hebei Key Laboratory for Chronic Disease, Tangshan Key Laboratory for Preclinical and Basic Research on Chronic Diseases, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063000, Hebei Province, China; 3Medical Central Laboratory, North China University of Science and Technology, Tangshan 063000, Hebei Province, China
  • Received:2017-10-19 Online:2017-12-08 Published:2018-01-04
  • Contact: Wang Zhi-qiang, Department of Orthopaedics, Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, Hebei Province, China
  • About author:Sun Fu-zhai, Studying for master’s degree, Department of Orthopaedics, Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, Hebei Province, China; Hebei Key Laboratory for Chronic Disease, Tangshan Key Laboratory for Preclinical and Basic Research on Chronic Diseases, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063000, Hebei Province, China
  • Supported by:
     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

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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.

Key words: Tantalum, Integrins, Biocompatible Materials, Tissue Engineering

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