Transfection of TGF-β shRNA by Using Ultrasound-targeted Microbubble Destruction to Inhibit the Early Adhesion Repair of Rats Wounded Achilles Tendon In vitro and In vivo.

Abstract

Tendon injury is a major orthopedic disorder. Ultrasound-targeted microbubble destruction (UTMD) provides a promising method for gene transfection, which can be used for the treatment of injured tendons. The purpose of this study was to investigate the optimal transforming growth factor beta (TGF-β) short hairpin RNA (shRNA) sequence and transfection conditions using UTMD in vitro and to identify its ability for inhibiting the early adhesion repair of rats wounded achilles tendons in vivo. The optimal sequence was selected analyzing under a fluorescence microscope and quantitative real-time reverse transcription polymerase chain reaction in vitro. In vivo, 40 rats with wounded Achilles tendons were divided into five groups: (1) control group, (2) plasmid group (3) plasmid + ultrasound group, (4) plasmid + microbubble group, (5) plasmid + microbubble + ultrasound group, and were euthanized at 14 days post treatment. TGF-β expression was evaluated using adhesion scores and pathological examinations. The optimal condition for UTMD delivery in vitro was 1W/cm2 of output intensity and a 30% duty cycle with 60 s irradiation time (P < 0.05). The transfection efficiency of the plasmid in group 5 was higher than that in other groups (P < 0.05). Moreover, the lowest adhesion index score and the least expression of TGF-β were shown in group 5 (P < 0.05). When compared with the other groups, group 5 had a milder inflammatory reaction. The results suggested that UTMD delivery of TGF-β shRNA offers a promising treatment approach for a tendon injury in vivo.