H-T Chen1, A-B Huang, Y-L He, J Bian, H-J Li. 1. Department of Orthopedics, Taizhou People's Hospital Affiliated to Nantong University, Taizhou, Jiangsu, China. handsome_lhj@139.com.
Abstract
OBJECTIVE: Our present study aimed to evaluate the effects of Wnt11 overexpression on the adipose-derived stem (ADSCs) cells differentiation to the nucleus pulposus (NP) cells and its function in the ADSCs cells growth, proliferation and induction of the NP cells markers. MATERIALS AND METHODS: The cell growth was detected using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT) assay and the cell cycle was assessed by the flow cytometry. The cells morphology was evaluated using the transmission electron microscopy. The transfection efficiencies of Wnt11 lentivirus were observed under fluorescence microscope. Besides, Quantitative Real-time PCR and Western blot analysis were applied to detect the relative mRNA and protein levels. RESULTS: Wnt11 lentivirus treatment could inhibit the ADSCs cells growth and arrest the cell cycle progression at the G0/G1 phase. Besides, the overexpression of Wnt11in ADSCs cells could induce the expression of the NP cells markers. Levels of SOX-9, aggrecan, and collagen type II were significantly increased in the ADSCs cells transfected with the Wnt11 lentivirus, in comparison with the untreated cells or the vector controls. CONCLUSIONS: The Wnt11 overexpression may provide some experimental evidence for the possible opportunity of the Wnt11 to promote the ADSCs cells differentiating to the NP cells. Therefore, the Wnt11 overexpression may have a potential utility for the treatment of the intervertebral disc degeneration.
OBJECTIVE: Our present study aimed to evaluate the effects of Wnt11 overexpression on the adipose-derived stem (ADSCs) cells differentiation to the nucleus pulposus (NP) cells and its function in the ADSCs cells growth, proliferation and induction of the NP cells markers. MATERIALS AND METHODS: The cell growth was detected using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT) assay and the cell cycle was assessed by the flow cytometry. The cells morphology was evaluated using the transmission electron microscopy. The transfection efficiencies of Wnt11 lentivirus were observed under fluorescence microscope. Besides, Quantitative Real-time PCR and Western blot analysis were applied to detect the relative mRNA and protein levels. RESULTS:Wnt11 lentivirus treatment could inhibit the ADSCs cells growth and arrest the cell cycle progression at the G0/G1 phase. Besides, the overexpression of Wnt11in ADSCs cells could induce the expression of the NP cells markers. Levels of SOX-9, aggrecan, and collagen type II were significantly increased in the ADSCs cells transfected with the Wnt11 lentivirus, in comparison with the untreated cells or the vector controls. CONCLUSIONS: The Wnt11 overexpression may provide some experimental evidence for the possible opportunity of the Wnt11 to promote the ADSCs cells differentiating to the NP cells. Therefore, the Wnt11 overexpression may have a potential utility for the treatment of the intervertebral disc degeneration.
Authors: Jingcai Wang; Min Gong; Shi Zuo; Jie Xu; Chris Paul; Hongxia Li; Min Liu; Yi-Gang Wang; Muhammad Ashraf; Meifeng Xu Journal: Genes (Basel) Date: 2020-10-29 Impact factor: 4.096