Effects of mechano-growth factor on proliferation,migration and differentiation of cartilage endplate-derived stem cells and their acting mechanisms
LI Haoxi
WU Chunlan
PENG Guanlu
MO Kaiqi
YAO Shuyu
YU Chengqiang
LI Zhuhai
WEI Jianxun
Abstract：Objective To explore the effects of mechano-growth factor(MGF)on proliferation,migration and differentiation of cartilage endplate-derived stem cells(CESCs)and their acting mechanisms.Methods The patients with degenerative disc disease who underwent disc fusion surgery in the People's Hospital of Guangxi Zhuang Autonomous Region were recruited.During the surgery,about 2 cm3 of the patients'cartilage endplate tissues of intervertebral disc were collected,and CESCs were collected and cultured.The effects of MGF on the proliferation of CESCs were evaluated by Cell Counting Kit-8(CCK-8)assay.The effects of MGF on the migration and invasion abilities of CESCs were evaluated by Transwell experiment.The expression levels of osteogenic,chondrogenic and lipogenic differentiation-related factors were detected by using reverse transcription-polymerase chain reaction(RT-PCR)and Western blot assay,and the influence of phosphorylated extracellular signal-regulated kinase(ERK)on the acting effect of MGF was analyzed.Results The results of CCK-8 assay showed that MGF could promote the proliferation of CESCs in a dose-dependent manner.The results of Transwell experiment showed that MGF could promote the migration of CESCs.After the intervention of ERK inhibitor PD98059,the proliferative effect of MGF was significantly weakened(P＜0.05).After the intervention of insulin-like growth factor-1 receptor(IGF-1R)inhibitor PQ401,the promoting migration effect of MGF was also significantly weakened(P＜0.05).The results of RT-PCR detection showed that the mRNA expression levels of alkaline phosphatase(ALP),Runt-related transcription factor 2(Runx2)and osteocalcin(OC)in the MGF group were significantly lower than those in the control group(P＜0.05),while the mRNA expression levels of aggrecan(AGG),SRY-related high-mobility group box gene-9(SOX-9)and type Ⅱ collagen(C Ⅱ)in the MGF group were significantly higher than those in the control group(P＜0.05).The results of Western blot assay also showed that the protein expression levels of ALP,Runx2 and OC in the MGF group were significantly lower than those in the control group(P＜0.05),while the protein expression levels of AGG,SOX-9 and C Ⅱ in the MGF group were significantly higher than those in the control group(P＜0.05).The ratio of phosphorylated ERK(p-ERK)to total extracellular signal-regulated kinase(t-ERK)in the MGF group was significantly higher than that in the control group(P＜0.05),and the ratio of p-ERK to t-ERK in the MGF+PQ401 group was lower than that in the MGF group,but the ratio of p-ERK to t-ERK in the MGF+PQ401 group was still higher than that in the control group,and the differences were statistically significant(P＜0.05).Conclusion MGF promotes the proliferation and migration of CESCs by inducing ERK phosphorylation.
Keywords：Cartilage endplate-derived stem cells(CESCs)Mechano-growth factor(MGF)ProliferationMigrationExtracellular signal-regulated kinase(ERK)Insulin-like growth factor-1 receptor(IGF-1 R)
Publication Date：2025-01-27
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 34-40 )