Inserm U872, Physiopathology of Ocular Diseases: Therapeutic Innovations, Paris, France.
Ultrasound Med Biol. 2011 Nov;37(11):1814-27. doi: 10.1016/j.ultrasmedbio.2011.07.010. Epub 2011 Oct 2.
This study aimed to assess application of ultrasound (US) combined with microbubbles (MB) to transfect the ciliary muscle of rat eyes. Reporter DNA plasmids encoding for Gaussia luciferase, β-galactosidase or the green fluorescent protein (GFP), alone or mixed with 50% Artison MB, were injected into the ciliary muscle, with or without US exposure (US set at 1 MHz, 2 W/cm(2), 50% duty cycle for 2 min). Luciferase activity was measured in ocular fluids at 7 and 30 days after sonoporation. At 1 week, the US+MB treatment showed a significant increase in luminescence compared with control eyes, injected with plasmid only, with or without MB (×2.6), and, reporter proteins were localized in the ciliary muscle by histochemical analysis. At 1 month, a significant decrease in luciferase activity was observed in all groups. A rise in lens and ciliary muscle temperature was measured during the procedure but did not result in any observable or microscopic damages at 1 and 8 days. The feasibility to transfer gene into the ciliary muscle by US and MB suggests that sonoporation may allow intraocular production of proteins for the treatment of inflammatory, angiogenic and/or degenerative retinal diseases.
本研究旨在评估超声（US）联合微泡（MB）转染大鼠眼睫状肌的应用。单独或混合 50% Artison MB 的报告 DNA 质粒，编码 Gaussia 荧光素酶、β-半乳糖苷酶或绿色荧光蛋白（GFP），被注射到睫状肌中，同时或不进行超声暴露（US 设置为 1 MHz、2 W/cm²、50%占空比 2 分钟）。在 sonoporation 后 7 天和 30 天测量眼液中的荧光素酶活性。在 1 周时，与仅注射质粒的对照眼相比，US+MB 处理的发光强度显著增加，无论是否有 MB（×2.6），并且组织化学分析显示报告蛋白定位于睫状肌。在 1 个月时，所有组的荧光素酶活性均显著下降。在该过程中测量了晶状体和睫状肌的温度升高，但在 1 天和 8 天时未导致任何可观察到或显微镜下的损伤。US 和 MB 将基因转移到睫状肌的可行性表明，声孔可以允许在眼内产生用于治疗炎症、血管生成和/或退行性视网膜疾病的蛋白质。
