Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (31): 4939-4944.doi: 10.12307/2021.133

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Effect of over expression of miR-378a on osteogenic and vascular differentiation of bone marrow mesenchymal stem cell sheet

Li Jun1, Zuo Xinhui1, Liu Xiaoyuan1, Zhang Kai1, Han Xiangzhen1, He Huiyu1, 2   

  1. 1Department of Prosthodontics, The First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China 

  • Received:2020-08-04 Revised:2020-08-07 Accepted:2020-09-05 Online:2021-11-08 Published:2021-04-25
  • Contact: He Huiyu, Chief physician, Professor, Doctoral supervisor, Master’s supervisor, Department of Prosthodontics, The First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Li Jun, Master candidate, Department of Prosthodontics, The First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    the Scientific and Technological Supporting Xinjiang Project of Xinjiang Uygur Autonomous Region, No. 2018E02060 (to HHY); the Postgraduate Scientific Research Innovation Project of Xinjiang Uygur Autonomous Region, No. XJ2020G198 (to LJ)

Abstract: BACKGROUND:  In recent years, the use of microRNAs to regulate the function and differentiation of stem cells is an attractive treatment model for regenerative medicine. Therefore, loading microRNA with stem cell sheet and to enhance its osteogenesis and angiogenesis properties is of great significance to develop a new type of repair material for bone defect.
OBJECTIVE: To investigate the effect of over expression of miR-378a gene on the osteogenic and angiogenic differentiation of bone marrow mesenchymal stem cell sheet.
METHODS:  Bone marrow mesenchymal stem cells from Sprague-Dawley rats were isolated and cultured. miR-378a lentivirus vector and negative control virus with appropriate titers were selected to infect bone marrow mesenchymal stem cells, which were recorded as LV-miR-378a-BMMSCs group and LV-BMMSCs group in turn, and cells not transfected with viral vector were used as blank group. Three groups of cells were inoculated into a six-well plate, and the membrane induced liquid was added for culture for 6 days. The morphology of the membrane was observed under the microscope. After the formation of the membrane, the membrane was cultured by osteogenic induction culture. At 3, 7, and 14 days of culture, the mRNA expressions of miR-378a, osteogenic gene runt-related transcription factor 2 (RUNX2) and type I collagen, angiogenesis genes vascular endothelial growth factor and platelet growth factor were detected by RT-PCR. Western blot assay was used to detect the protein expression of RUNX2 and vascular endothelial growth factor when cultured for 14 days.  
RESULTS AND CONCLUSION: (1) On day 2 after the induction of membrane formation, a small amount of stroma was observed to be connected between the central spindle cells of the membrane microscopically, and the edge of the membrane was furred and discontinuous. On day 6, the central cells of the membrane were overlapped and dense, and a large number of cell matrix networks were cross-connected. The edges of the membrane showed continuous and widened folds and rolled toward the center, showing no significant difference from the morphological changes of the blank group. (2) The mRNA expressions of miR-378a, RUNX2, type I collagen, vascular endothelial growth factor and platelet growth factor at each time points in the LV-miR-378a-BMMSCs group were higher than those in the LV-BMMSCs group and the blank group (P < 0.001), and the protein expressions of RUNX2 and vascular endothelial growth factor were all higher than those in LV-BMMSCs group (P < 0.001) and blank group (P < 0.01). (3) The results showed that overexpression of miR-378a promoted the osteogenesis and angiogenesis of the bone marrow mesenchymal stem cell sheet in vitro.

Key words: bone, material, miR-378a, bone marrow mesenchymal stem cell, sheet, osteogenic differentiation, angiogenic differentiation, bone tissue engineering

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