Effect of curcumin-primed human bone marrow mesenchymal stem cells-derived exosomes on osteo-arthritic chondrocytes
Objective To observe the effect of curcumin-primed exosomes derived from human bone marrow mesenchymal stem cells(hBMSCs)on osteoarthritic chondrocyte proliferation,apoptosis and expression of key genes and proteins.Methods A total of 10 patients who were hospitalized from March 2023 to June 2023 were selected from the Department of Orthopedic Surgery of the First Affiliated Hospital of Zhengzhou University,and hBMSCs and osteoarthritic chondrocytes were extracted and cultured from 10 patients undergoing joint replacement,including 5 patients with fracture of neck of femur,aged(56.8±6.0),60％females,and 5 patients with osteoarthritis of the knee,aged(54.6±5.9),80％females.After adding 10 µmol/L curcumin to the BMSCs culture medium for 48 h,curcumin exo-somes were extracted from the culture supernatants of the BMSCs using differential centrifugation and ultra-centrifugation(100 000 ×g centrifugation for 70 min).Osteoarthritic chondrocytes were categorized into control group,interleukin(IL)-1 β group(1 ng/ml IL-1 β pre-stimulation),IL-1β+curcumin group(1 ng/ml IL-1 β pre-stimulation+free curcumin)and IL-1 β+curcumin-exosomes group(1 ng/ml IL-1 βpre-stimulation+curcumin-primed hBMSC-derived exosomes),and the effect of curcumin-primed hBMSC-derived exosomes on the proliferation and apoptosis of osteoarthritic chondrocytes was detected by methyl thiazolyl tetrazolium(MTT)assay,cysteinyl aspartate-specific protease(Caspase)-3/7 apoptosis as-say,respectively.Real-time fluorescent quantitative polymerase chain reaction(RT-PCR)and Western blotting were used to detect differential expression of key genes and proteins.Comparison of data above the two groups was performed by one-way ANOVA.Results The extracted exosomes were round membranous vesicles with a particle size of about 90-120 nm,and expressed exosome-specific markers CD9,CD63,indicating successful characterization.Exosomes labeled by red fluorescence within chondrocytes were visi-ble under fluorescence microscopy,indicating that exosomes were taken up by chondrocytes.The results of MTT method showed that the control group was set to 1.00,IL-1β group(0.50±0.03),IL-1β+curcumin group(0.51±0.04)and IL-1 β+curcumin-exosome group(0.83±0.03).The proliferative capacity of the IL-1 β+curcumin-exosomes group was significantly higher than that of the IL-1 β group(0.83±0.03 vs.0.50±0.03,t=8.77,P＜0.001).Caspase-3/7 activity apoptosis assay showed that the control group was set to 1.00,and the IL-1 β group(1.53±0.08),the IL-1 β+curcumin group(1.51±0.05),and IL-1 β+curcumin exosome group(0.73±0.04).The apoptosis rate in the IL-1 β+curcumin-exosomes group was significantly lower than that in the IL-1β group(0.73±0.04 vs.1.53±0.08,t=11.89,P＜0.01).RT-PCR results showed that IL-6 and MMP-13 expression in the IL-1β+curcumin exosome group was significantly lower than that in the IL-1β group(0.59±0.04,0.39±0.03 vs.1.00,0.68±0.04,0.40±0.04 vs.1.00,set at 1.00 in IL-1 β group,t=25.24、22.11,P＜0.01),COL2A1 expression was significantly higher than that in IL-1β group(2.39±0.17,2.91±0.17 vs.1.00,t=16.19,P＜0.01),and IL-1β+curcumin exosome group had lower expression of IL-6,MMP-13 and higher expression of COL2A1 that IL-1 β+curcumin group.Conclusion Curcumin-primed hBMSC-derived exosomes enhance osteoarthritic chondrocyte proliferation,reduce apoptosis,attenuate cartilage matrix decomposition,and promote cartilage repair.
Bone marrow mesenchymal stem cellsExosomesCurcuminChondrocytesOsteoarthritis
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