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
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
复制标题
超声靶向微泡破坏转染TGF-β shRNA抑制大鼠跟腱损伤体内外早期粘连修复
DOI：
10.2174/1566523220666200516165828
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发表时间：
2020-01-01
影响因子：
3.6
通讯作者：
Tang, Xinyi
中科院分区：
文献类型：
--
作者：
Huang, Songya;Xiang, Xi;Tang, Xinyi
背景：肌腱损伤是一种主要的骨科疾病。超声靶向微泡破坏法（Ultrasound-targeted microbubble destruction，UTMD）为基因转染提供了一种很有前途的方法，可用于肌腱损伤的治疗。本研究的目的是探讨最佳转化生长因子β（TGF-β）短发夹RNA（shRNA）序列和转染条件，并鉴定其抑制大鼠跟腱损伤后早期粘连修复的能力。vivo.Methods:荧光显微镜下分析和体外实时定量逆转录聚合酶链反应筛选出最佳序列。在体实验中，将40只跟腱损伤大鼠随机分为5组：（1）对照组，（2）质粒组，（3）质粒+超声组，（4）质粒+微泡组，（5）质粒+微泡+超声组，于治疗后14 d处死。结果：UTMD体外释放的最佳条件为输出功率1 W/cm 2，占空比30%，照射时间60 s（P < 0.05）。第5组质粒转染效率高于其他各组（P < 0.05）。第5组粘附指数评分最低，TGF-β表达最少（P < 0.05）。与其他组相比，第5组有较轻的炎症反应。结论：结果表明，UTMD交付TGF-β shRNA提供了一个有前途的治疗方法，在体内肌腱损伤。
Background: 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.Objective: The purpose of this study was to investigate the optimal transforming growth factor beta (TGF-beta) 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.Methods: 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-beta expression was evaluated using adhesion scores and pathological examinations.Results: The optimal condition for UTMD delivery in vitro was 1W/cm(2) 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-beta were shown in group 5 (P < 0.05). When compared with the other groups, group 5 had a milder inflammatory reaction.Conclusion: The results suggested that UTMD delivery of TGF-beta shRNA offers a promising treatment approach for a tendon injury in vivo.