Skeletal Biotech Laboratory, The Hebrew University-Hadassah Faculty of Dental Medicine, Ein Kerem, Jerusalem 91120, Israel; Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Department of Surgery, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA; Department of Orthopedics, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
Mol Ther. 2018 Jul 5;26(7):1746-1755. doi: 10.1016/j.ymthe.2018.04.020. Epub 2018 Apr 26.
Ligament injuries occur frequently, substantially hindering routine daily activities and sports participation in patients. Surgical reconstruction using autogenous or allogeneic tissues is the gold standard treatment for ligament injuries. Although surgeons routinely perform ligament reconstructions, the integrity of these reconstructions largely depends on adequate biological healing of the interface between the ligament graft and the bone. We hypothesized that localized ultrasound-mediated, microbubble-enhanced therapeutic gene delivery to endogenous stem cells would lead to significantly improved ligament graft integration. To test this hypothesis, an anterior cruciate ligament reconstruction procedure was performed in Yucatan mini-pigs. A collagen scaffold was implanted in the reconstruction sites to facilitate recruitment of endogenous mesenchymal stem cells. Ultrasound-mediated reporter gene delivery successfully transfected 40% of cells recruited to the reconstruction sites. When BMP-6 encoding DNA was delivered, BMP-6 expression in the reconstruction sites was significantly enhanced. Micro-computed tomography and biomechanical analyses showed that ultrasound-mediated BMP-6 gene delivery led to significantly enhanced osteointegration in all animals 8 weeks after surgery. Collectively, these findings demonstrate that ultrasound-mediated gene delivery to endogenous mesenchymal progenitor cells can effectively improve ligament reconstruction in large animals, thereby addressing a major unmet orthopedic need and offering new possibilities for translation to the clinical setting.
韧带损伤较为常见，严重影响患者的日常活动和运动参与。使用自体或同种异体组织进行手术重建是治疗韧带损伤的金标准。尽管外科医生经常进行韧带重建，但这些重建的完整性在很大程度上取决于韧带移植物与骨骼之间界面的充分生物愈合。我们假设局部超声介导的微泡增强治疗性基因传递到内源性干细胞将导致韧带移植物的整合显著改善。为了验证这一假设，在尤卡坦小型猪中进行了前交叉韧带重建手术。在重建部位植入胶原支架，以促进内源性间充质干细胞的募集。超声介导的报告基因传递成功转染了募集到重建部位的 40%的细胞。当传递编码 BMP-6 的 DNA 时，重建部位的 BMP-6 表达显著增强。微计算机断层扫描和生物力学分析表明，超声介导的 BMP-6 基因传递可导致所有动物在手术后 8 周时显著增强骨整合。总之，这些发现表明，将基因传递到内源性间充质祖细胞的超声介导可以有效地改善大动物的韧带重建，从而解决主要的未满足的骨科需求，并为转化为临床环境提供新的可能性。
