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Ultrasound-targeted microbubble destruction (UTMD) is a novel method for gene transfection. The aim of the present study was to identify the most suitable method of tumor necrosis factor (TNF)-α receptor (TNFR) gene transfection using UTMD for systemically treating a rat model of collagen-induced arthritis (CIA). Plasmids encoding the TNFR and enhanced green fluorescent protein (EGFP) with or without microbubbles were locally injected into the skeletal muscle and synovial membrane of CIA rats. The rats were divided into the following 6 groups: i) Group 1, plasmid + microbubble + ultrasound (muscle group); ii) group 2, plasmid + microbubble + ultrasound (joint group); iii) group 3, plasmid + ultrasound; iv) group 4, plasmid + microbubble; v) group 5, plasmid only and; vi) group 6, untreated controls. Rats were sacrificed at 2, 4 and 8 weeks of treatment. The transfection efficiency of the plasmids in the muscle or synovium was observed by fluorescence microscopy. Arthritis scores were calculated and serum levels of TNF-α were measured prior to and following treatment. Bilateral ankle joints were obtained and stained to observe synovial inflammation and the expression of TNF-α. EGFP expression was detected in all treated groups at each time point, and the fluorescence intensity of groups 1 and 2 was significantly greater than that of the other groups (P&lt;0.05). For groups 1 and 2, the reductions in joint scores and serum levels of TNF-α were significant compared with the other groups (P&lt;0.05). The number of synovial inflammatory cells and the synovial expression of TNF-α presented similar results among all experimental groups and no significant difference was observed between groups 1 and 2. Therefore, the results of the present study suggest that UTMD significantly enhanced the efficiency of TNFR gene transfection in the muscle and inflamed synovium of rats with. Regardless of whether the transfected TNFR gene was injected into the muscle or joint, it was continuously expressed in the rats for at least 8 weeks, which may improve arthritic symptoms and reduce the levels of inflammatory factors in the synovial tissues and peripheral blood.

experimental and therapeutic medicine

Experimental study of TNF‑α receptor gene transfection by ultrasound‑targeted microbubble destruction to treat collagen‑induced arthritis in rats in�vivo