| Objective: To explore whether the effect of microbubble cavitation with ultrasound can reduce or block the angiogenesis of synovial,thus playing a role in the treatment of rheumatoid arthritis,and hope to provide a new idea and theoretical basis for clinical treatment.Methods:1.We made the collagen-induced arthritis(CIA)models from 100 SD rats with chicken type II collagen and incomplate adjuvant.After modeled,the 72 models were successfully made,and then randomly divided them into 4 groups.Group1:microbubbles + ultrasound;Group 2: saline + ultrasound;Group 3: ultrasound;Group 4: control group.Applied couplant to the hind paws of rats on the 14 th day after the initial immunization,rats in Group 1 were injected with 1ml microbubble contrast agent via tail vein,set the MI to be 1.0,and then irradiated both of the hind paws by ultrasound for 3 minutes immediately.Rats in Group 2 were injected with1 ml saline contrast agent and were irradiated on both of the hind paws by ultrasound for 3 minutes immediately under the same condition.Rats in Group 3 were irradiated by ultrasound for 3 minutes.Rats in Group 4 did not had any treatment.Fed each rat in single cage after all treatments.2.We scored the arthritis index(AI)every 7 days using a five-point scale according to the extent and deformation of the joints in rats for 4 weeks.3.High-frequency color Doppler ultrasonography of the ankle joints was observed every 7 days for 4 consecutive weeks.The condition of the synovial membrane was observed,and the thickness of the thickest part of the synovial membrane was measured.The mean value of the three records was measured.The blood supply of the synovial membrane was observed by color Doppler ultrasonography for 4 consecutive weeks.The blood flow signal in the synovial membrane was divided into 4 levels and recorded according to the semi-quantitative method.4.Rats in each group was injected with SonoVue via tail vein every 7 days for 4consecutive weeks,and the hind paw of each rat was observed by contrast.The lesion synovium was selected as the Region of Interest(ROI).The machine’s own quantitative analysis software performs time-intensity curve(TIC)analysis,and obtains contrast-enhanced perfusion parameters,which record peak(Peak),regional bleed volume(RBV),and regional bleed flow(RBF).5.We sacrificed the rats at the end of the last week.The synovial tissue of rats was fixed in 10% neutral formalin for 24 hours,embedded in paraffin,and observed the synovial tissue of rats and evaluated the neovascularization of synovial tissue.Then analyzed VEGF and CD34 expression and calculated the MVD.6.Statistical analysis:Measurement data were analyzed by Non-parmetric test for independent samples,count data were expressed as mean ± standard devietion,and analyzed by one-way ANOVA,using the SPSS version 20.0 software.p < 0.05 was considered as statistically significant.Results:1.AI: The arthritis index scores increased from the first week to the third week after treatment,peaked at the third week,and decreased at the fourth week.However,the AI of synovial membrane in group 1 at each observation time point was significantly smaller than that in the other three groups(p<0.05);Group 2,Group 3and Group 4 were compared with each other,there was no significant difference.2.Synovial thickness: At the first week after treatment,the incidence of arthritis reached a peak,and the synovial thickening was obvious.With the prolongation of time,the inflammatory reaction gradually increased,and the synovial membrane gradually thickened.At the 3rd week,the synovial thickening reached a peak.Synovial swelling was relieved at the 4th week.However,the thickness of synovial membrane in group 1 at each observation time point was significantly smaller than that in the other three groups.At the 1st,2nd,3rd,and 4th week after treatment,the differences between the Group 1 with the Group 2,Group 3 and Group 4 were significant(p<0.05);Group 2,Group 3 and Group 4 were compared with each other,there was no significant difference.3.CDFI: The blood circulation gradually increased from the first week to the2 nd week after treatment,and there was no obvious peak trend at the 3rd week,then decreased at the 4th week.However,the blood supply grading of group 1 was lower than the other 3 groups at each observation time point.At the 1st,2nd,3rd,and 4th week after treatment,the differences between the Group 1 with the Group 2,Group 3and Group 4 were significant(p<0.05);Group 2,Group 3 and Group 4 were compared with each other,there was no significant difference.4.Peak,RBV and RBF gradually increased from 1st week to the 2nd week after treatment,and all parameters peaked at the 3rd week after treatment,and decreased at the 4th week.However,the values of group 1 were lower than the other 3 groups at each observation time point.At the 1st,2nd,3rd,and 4th week after treatment,the differences between the Group 1 with the Group 2,Group 3 and Group 4 were significant(p<0.05);Group 2,Group 3 and Group 4 were compared with each other,there was no significant difference.5.Immunohistochemistry: The expression of VEGF and MVD differences between the Group 1 with the Group 2,Group 3 and Group 4 were significant(p<0.05);Group 2,Group 3 and Group 4 were compared with each other,there was no significant difference.Conclusions:1.Microbubble cavitation with ultrasound can be used to treat rheumatoid arthritis in rats.2.Microbubble cavitation with ultrasound destroyed the synovial angiogenesis and thus played a role in the treatment of rheumatoid arthritis in rats.3.The rheumatoid arthritis of CIA model will show a self-healing after the inflammation developed to a peak without any treatment. |
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