Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (19): 2985-2990.doi: 10.3969/j.issn.2095-4344.2042

Previous Articles     Next Articles

Zinc ion concentration affects proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells 

Zhao Qiao1, 2, Yang Fei3, Zhang Chengdong2, Chen Shuo2, Sun Xiao2, Zhang Bo3, Xiao Dongqin2, Liu Kang2, Feng Gang1, 2, Duan Ke1   

  1. 1Department of Orthopedics and Arthrology, Affiliated Hospital of Southwest Medical University, Laboratory of Research and Development and Application of Orthopedic Devices; 2Second Clinical College of North Sichuan Medical College • Tissue Engineering and Stem Cell Research Institute of Nanchong Central Hospital; 3Department of Orthopedics, Nanchong Central Hospital
  • Received:2019-05-08 Revised:2019-05-22 Accepted:2019-08-07 Online:2020-07-08 Published:2020-04-07
  • Contact: Feng Gang, MD, Professor, Department of Orthopedics and Arthrology, Affiliated Hospital of Southwest Medical University, Laboratory of Research and Development and Application of Orthopedic Devices, Luzhou 646000, Sichuan Province, China; Second Clinical College of North Sichuan Medical College • Tissue Engineering and Stem Cell Research Institute of Nanchong Central Hospital, Nanchong 637000, Sichuan Province, China Duan Ke, MD, Associate professor, Department of Orthopedics and Arthrology, Affiliated Hospital of Southwest Medical University, Laboratory of Research and Development and Application of Orthopedic Devices, Luzhou 646000, Sichuan Province, China
  • About author:Zhao Qiao, Master candidate, Department of Orthopedics and Arthrology, Affiliated Hospital of Southwest Medical University, Laboratory of Research and Development and Application of Orthopedic Devices, Luzhou 646000, Sichuan Province, China; Second Clinical College of North Sichuan Medical College • Tissue Engineering and Stem Cell Research Institute of Nanchong Central Hospital, Nanchong 637000, Sichuan Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81171472; the Applied Foundation Project of Science and Technology Department of Sichuan Province, No. 2016JY0123, 2018JY0100; the Scientific Research Project of Sichuan Health and Family Planning Commission, No. 18PJ476; the Scientific Research Project of Sichuan Provincial Department of Education, No. 17ZA0178; the Nanchong City-School Cooperative Research Special Funds, No. NSMC20170310, NSMC20170453, 18SXHZ0374, 18SXHZ0377, 18SXHZ0124

PDF

137

Abstract:

BACKGROUND: Zinc ion is a necessary metal trace element in human body, which plays an important role in inhibiting osteoclast activity and promoting osteogenic differentiation. However, the effect of zinc ion at different concentrations on osteogenic differentiation of rabbit bone marrow mesenchymal stem cells has been rarely studied.

OBJECTIVE: To evaluate the effects of zinc ion at different concentrations on the proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells for exploring the appropriate zinc ion concentration.  

METHODS: Bone marrow mesenchymal stem cells were extracted from the long bone marrow of rabbits and passaged. The cells were divided into six groups, including 10-4, 10-5, 10-6, 10-7 and 10-8 mol/L zinc ion groups, and blank control group. The cell proliferation activity was measured by Cell Counting Kit-8 and the growth curve was drawn. Alkaline Phosphatase Assay kit was used to evaluate the osteogenic differentiation ability of the cells, and the growth activity of the cells was shown by Live-Dead Cell Staining Kit. The mineralization ability of the cells was observed by alizarin red S staining. The expression of osteogenic genes was detected by fluorescence quantitative polymerase chain reaction.

RESULTS AND CONCLUSION: The number of cells in each group showed an increasing trend with the prolongation of time except for the 10-4 mol/L zinc ion group. After 5 days of culture, the number of cells was highest in the 10-7 mol/L zinc ion group, but lowest in the 10-4 mol/L zinc ion group. Alkaline phosphatase activity was highest when zinc ion concentration was 10-7 mol/L, but lowest at the 10-4 mol/L zinc ion after cell culture for 14 days. The method of Live-Dead Cell Staining revealed the number of viable cells was highest in the 10-7 mol/L zinc ion group. All cells were almost dead in the 10-4 mol/L zinc ion group. Calcium nodule formation was visible, and the expression level of osteogenic genes was highest in the 10-7 mol/L zinc ion group after 14 days of cell culture. These findings suggest that zinc ion can effectively promote the proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells in a certain concentration range. The promoting effects of zinc ion on proliferation, osteogenic induction and mineralization are strongest. Therefore, the addition of a suitable concentration of zinc ions in the medium is beneficial for the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells.

Key words: bone marrow mesenchymal stem cells, zinc ion, cell proliferation, osteogenic differentiation, mineralization

CLC Number: