Gujian(骨健)peroral liquids combined with mesenchymal stem cell-derived exosome against knee osteoarthri-tis in mice:an experimental study
Objective:To observe the effects of Gujian(骨健,GJ)peroral liquids combined with mesenchymal stem cell-derived exo-some(MSC-EXO)on knee osteoarthritis(KOA)in mice,and to explore its mechanism.Methods:Twenty-five 12-week-old specific pathogen free(SPF)-grade male C57BL/6J mice were randomized into sham-operated group,model group,GJ group,MSC-EXO group,GJ combined MSC-EXO group,5 cases in each group.All mice but the ones in sham-operated group were subjected to destabilized medial meniscus(DMM)method on the right knees for inducing KOA,while the ones in sham-operated group underwent surgery for merely incising the joint capsule,but without transecting medial meniscus.After successful modeling,the mice in GJ group and GJ combined MSC-EXO group were intervened by intragastric administration with 0.2 mL GJ peroral liquids,while the ones in sham-operated group,model group and MSC-EXO group with the same dose of normal saline,once a day for consecutive 8 weeks;moreover,the mice in MSC-EXO group and GJ combined MSC-EXO group were further intervened by knee intra-articular injection of 10 μL MSC-EXO,while the ones in sham-operated group,model group and GJ group by the same dose of phosphate buffer solution(PBS),once a week for consecutive 8 weeks.After the end of interven-tion,the mice were sacrificed by cervical dislocation method,and their right knees were harvested for scanning and analysing by using Mi-cro-CT,followed by observation on histopathological changes of knee tissues and determination of the expression level of collagen Ⅱ α1(Col2A1)in knee cartilage tissues.Results:①The bone volume fraction was lower in sham-operated group,GJ group,MSC-EXO group and GJ combined MSC-EXO group compared to model group(P=0.004,P=0.007,P=0.040,P=0.001),while,there was no statistical difference between GJ group and GJ combined MSC-EXO group as well as between MSC-EXO group and GJ combined MSC-EXO group(P=0.108,P=0.126).The trabecular thickness was greater in sham-operated group,GJ group,MSC-EXO group and GJ combined MSC-EXO group compared to model group(P=0.015,P=0.023,P=0.023,P=0.009),while,there was no statistical difference between GJ group and GJ combined MSC-EXO group as well as between MSC-EXO group and GJ combined MSC-EXO group(P=0.347,P=0.062).The trabecular number was more in sham-operated group and GJ group compared to model group(P=0.016,P=0.021),while,there was no statistical difference between MSC-EXO group and model group as well as between GJ combined MSC-EXO group and model group(P=0.196,P=0.206).The trabecular separation was lower in sham-operated group,GJ group,MSC-EXO group and GJ combined MSC-EXO group compared to model group(P=0.016,P=0.012,P=0.007,P=0.001),and was lower in GJ combined MSC-EXO group compared to GJ group(P=0.016),while,there was no statistical difference between MSC-EXO group and GJ combined MSC-EXO group(P=0.224).②The mice in sham-operated group presented with smooth articular cartilage surface and neatly arranged chondrocytes,which sug-gesting no obvious degeneration in articular cartilage;compared with that of sham-operated group,the changes,manifesting as obviously worn articular cartilage tissues,decreased and disordered arranged chondrocytes,were observed in mice of model group;compared with that of model grouop,the mice in GJ group and MSC-EXO group presented with partial worn articular cartilage with increased and neatly arranged chondrocytes;the improvement was better in GJ combined MSC-EXO group in contrast to GJ group and MSC-EXO group.③The positive ex-pression area ratio of Col2A1 in knee cartilage tissues was greater in mice of sham-operated group,GJ group,MSC-EXO group and GJ com-bined MSC-EXO group compared to model group(P=0.000,P=0.000,P=0.000,P=0.000),and was greater in GJ combined MSC-EXO group compared to GJ group and MSC-EXO group(P=0.000,P=0.000).Conclusion:GJ peroral liquids combined with MSC-EXO can improve the pathological state of articular cartilage and the abnormal microstructure of subchondral bone in KOA mice,and it behaves a synergistic therapeutic action.It may exert the effects by up-regulating the expression of Col2A1 in knee cartilage tissues.
万方数据
维普
