Résumé
BACKGROUND:As a member of bone morphogenetic proteins,growth differentiation factor-5 shows promising potential in the application of cartilage and bone repair.The affinity of growth differentiation factor-5 onto bone tissue determines protein use efficiency,so it is of great significance to prepare growth differentiation factor-5 with bone targeting capability. OBJECTIVE:To modify growth differentiation factor-5 using bisphosphonates and investigate the effects of modified protein on the growth of preosteoblasts in mice. METHODS:Pamidronate disodium/growth differentiation factor-5 complex was prepared using chemical crosslinking to couple growth differentiation factor-5 with pamidronate disodium.The functional groups and structures of the complex were characterized using Fourier transform infrared spectroscopy and circular dichromatography.To determine the bone targeting in vitro,the binding of the modified growth differentiation factor-5 with calcium phosphate and in vitro release amount of growth differentiation factor-5 were measured with an ELISA kit.Growth differentiation factor-5(control group)and the pamidronate disodium/growth differentiation factor-5 complex(experimental group)were co-cultured with preosteoblasts MC3T3-E1.Individually cultured cells were blank controls.The effect of the complex on cell proliferation and differentiation was evaluated. RESULTS AND CONCLUSION:(1)The infrared spectroscopy and circular dichromatography results indicated that the bisphosphonate/growth differentiation factor-5 complex was successfully prepared without significant changes in the protein secondary structure.In vitro protein adsorption results showed that growth differentiation factor-5 adsorption on calcium phosphate was increased by about one time after coupling with a bisphosphonate.In the presence of cysteine,growth differentiation factor-5 could be released from the bisphosphonate/growth differentiation factor-5 complex.(2)CCK-8 assay results showed that the absorbance value of the experimental group cultured for 4 and 7 days was higher than that of the control group and blank control group(P<0.000 1).After 7 days of culture,the expression of alkaline phosphatase in the experimental group was significantly higher than that in the control group and blank control group(P<0.000 1).After 13 days of culture,the content of calcium nodules in the experimental group was significantly higher than that in the control group and the blank control group(P<0.000 1).The results of qRT-PCR showed that the mRNA expression of alkaline phosphatase,osteocalcin and Runx2 in the experimental group was higher than that in the control group and the blank control group after 7 days of culture(P<0.01,P<0.001,P<0.000 1).(3)These findings exhibit that bisphosphonate modification can enhance the binding capacity of growth differentiation factor-5 to calcium phosphate as well as improve its biological activity.
Résumé
Objective:To explore the effect of extracorporeal shock wave combined with meloxicam tablets on joint function and serum matrix metalloproteinase-9 (MMP-9) and resistin levels in elderly patients with osteoarthritis.Methods:80 cases of elderly patients with osteoarthritis in Nanchong Central Hospital from May 2016 to March 2019 were selected as the research objects, and they were randomly divided into two groups, 40 cases in each group. The control group was given conventional oral medicine (meloxicam + potassium glucosamine sulfate tablets), and the observation group was given meloxicam tablets combined with extracorporeal shock wave therapy. The total effective rate, incidence of adverse reactions, and the scores of osteoarthritis symptoms (WOMCA), joint function (Lysholm), serum MMP-9 and resistin before and after 8 weeks' treatment were compared between the two groups.Results:After treatment, the WOMCA score of the observation group was lower than that of the control group, and Lysholm score was higher than that of the control group ( P<0.05); the total effective rate of the observation group (95.00%) was higher than of the control group (77.50%) ( P<0.05); the serum resistin and MMP-9 levels of the observation group were lower than those of the control group after treatment ( P<0.05); there was no significant difference in the incidence of adverse reactions between the observation group (15.00%) and the control group (17.50%) ( P>0.05). Conclusions:Extracorporeal shock wave combined with meloxicam tablets in the treatment of elderly patients with osteoarthritis can significantly improve the clinical symptoms, promote the recovery of joint function, reduce the levels of serum resistin and MMP-9, and further improve the treatment effect with high safety.