Effect Of Warm Acupuncture Combined With Bone Marrow Mesenchymal Stem Cells Transplantation On Cartilage Tissue In Rabbit Knee Osteoarthritis

ObjectiveIn this experiment,warm acupuncture combined with bone marrow mesenchymal stem cells(BMSCs)transplantation was used to treat knee osteoarthritis(KOA)of rabbits.The study aimed to investigate the protective effect of warm acupuncture combined with BMSCs transplantation on cartilage tissue injury in rabbits KOA as well as the underlying mechanism.Methods50 rabbits(male/female ratio,1:1)were randomly divided into 5 groups: blank group,model group,warm acupuncture group,BMSCs group,warm acupuncture combined with BMSCs group,10 rabbits in each group.In addition to the blank group,the other 4 groups were fixed in the extended position of the right hind limb for 6 weeks to replicate the rabbit model of medium-term KOA cartilage degeneration,and the model was successfully constructed by X-ray examination.After the model was successfully constructed,the corresponding intervention measures were given respectively.Modified LequesneMG knee joint assessment scale used to evaluate the degree of knee joint behavior,tai-ping peng method generally observed KOA rabbit cartilage histomorphology changes,hematoxylin eosin staining,safranine O-green FCF staining,toluidine blue staining of KOA rabbit cartilage tissue pathology observation,transmission electron microscope observed the cartilage cell ultrastructure and apoptotic body,TUNEL staining observed the cartilage cell apoptosis,immunohistochemical and qPCR observed cartilage tissue apoptosis-related factors Bax,Bcl-2,Caspase-3 expression.SPSS 20.0 statistical software was used to analyze the final experimental values.Results 1.Protective effect of warm acupuncture combined with BMSCs on KOA cartilage tissue injury in rabbitsCompared with the blank group,model group of rabbit knee joint swelling,range decreased significantly,LequesneMG score increased significantly(P < 0.01).Cartilage tissue pathological damage was severe,HE staining Mankin’s score significantly higher(P < 0.01).Safranine O-green FCF staining and toluidine blue staining observed widespread loss of dye,transmission electron microscopy observed the degeneration of cartilaginous ultrastructure,the collapse of extracellular blood vessels,the visible apoptotic body locally.Compared with model group,the joint function was recovered,the symptoms were relieved and LequesneMG score was significantly decreased(P<0.01)in warm acupuncture group,BMSCs group and warm acupuncture combined BMSCs group.In addition,cartilage tissue pathological damage degree was improved(P<0.01),the degeneration of cartilaginous ultrastructure were recovered,peripheral vascular was slightly edema,blood supply gradually restored,even a small amount of red blood cells found.Furthermore,the above effects of warm acupuncture combined with BMSCs was better than that of warm acupuncture and BMSCs transplantation alone(P<0.05).2.Protective mechanism of warm acupuncture combined with BMSCs on KOA cartilage tissue injury in rabbitsCompared with the blank group,the number of TUNEL positive chondrocytes in the model group was significantly increased(P<0.01),and the expression of Bax,Caspase-3 protein and mRNA in cartilage tissue was significantly increased(P<0.01),while the expression of Bcl-2 protein and mRNA was decreased(P<0.01).Compared with the model group,warm acupuncture combined with BMSCs transplantation significantly reduced the number of chondrocytes apoptosis(P<0.01),and the expression of apoptosis-related factors in cartilage tissue was significantly reversed,in which the expression of Bcl-2 protein and mRNA was significantly increased,and the expression of Bax and Caspase-3 protein and mRNA was significantly decreased(P<0.01).Conclusion 1.Warm acupuncture combined with BMSCs transplantation could effectively inhibit KOA cartilage tissue injury in rabbits.The effect was significantly better than that of warm acupuncture alone and BMSCs alone,confirming the synergistic effect.2.Warm acupuncture combined with BMSCs transplantation was benefical to reducing cartilage tissue injury may be by reducing the expression of Bax and Caspase-3,elevating the level of Bcl-2,and inhibiting chondrocyte apoptosis on KOA in rabbits.