Duke University School of Medicine, Durham, NC, USA.
Exp Eye Res. 2010 Apr;90(4):501-6. doi: 10.1016/j.exer.2010.01.001. Epub 2010 Jan 11.
We quantified episcleral drug clearance of sodium fluorescein (NaFl) in rats to examine the hypothesis that there is rapid clearance of episcleral water soluble drugs, and that this rapid clearance may limit the amount of drug that is able to reach the posterior segment from an episcleral location. 2 mm implants containing either 12 or 22 microg of NaFl were manufactured and in vitro release rates were determined. Implants were placed in the sub-Tenon's space and the amount of drug remaining in the conjunctiva/sclera/choroid complex (CSCC) at various time points was quantified following tissue solubilization and fluorescence quantification using a spectrofluorometer. Kinetics of NaFl clearance was determined in live animals, following euthanasia and in animals in which choroidal non-perfusion had been achieved with indocyanine green-enhanced 810 nm diode laser thrombosis of the choroidal vasculature. Choroidal non-perfusion in these laser-treated rats was verified with Concavalin-A staining of choroidal flatmounts. In vitro, >99% of drug was released by 25 min for the low dose implants, and by 60 min for the high dose implants. In vivo, both implant doses were >99% cleared from the episcleral tissue by 3 h. By 7 h, an average of only 0.14 +/- 0.131 ng of NaFl per mg of wet tissue weight (mean +/- SD) remained in the CSCC with the low dose implant, and 0.29 +/- 0.428 ng of NaFl per mg of wet tissue weight remained in animals with the high dose implant. By comparison, in euthanized animals at 7 h following sub-Tenon's implantation, 432.0 +/- 181.40 ng of NaFl per mg of wet tissue weight was in the episcleral tissue of animals with the low dose implant, and of 787.8 +/- 409.89 ng of NaFl per mg of wet tissue weight remained in the animals with the high dose implant. In live animals with selective thrombosis of the choroidal vasculature, the difference in the amount of drug remaining in the episcleral tissue as compared to control live animals was not significant at all time points for both implant doses. In conclusion, there is rapid clearance of episcleral NaFl delivered from a bioerodible sub-tenon's implant. The clearance mechanisms are dramatically reduced following euthanasia, suggesting that elimination is occurring via active physiologic mechanisms, rather than by passive diffusion clearance (CL(diff)) (Pfister et al., 2003). Interestingly, the choroid does not appear to play a prominent role as clearance of episcleral NaFl was not affected by elimination of choroidal blood flow. Further work is needed to delineate the pathways of episcleral drug clearance.
我们定量研究了巩膜外的药物清除率,以检验这样一个假设:巩膜外水溶性药物清除速度很快,而这种快速清除可能限制了药物从巩膜位置到达后节的量。我们制造了含有 12 或 22 微克 NaFl 的 2 毫米植入物,并确定了体外释放率。植入物被放置在 Tenon 下腔,在组织溶解后,通过荧光分光光度计定量检测结膜/巩膜/脉络膜复合体 (CSCC) 中残留的药物量。在活动物中,通过安乐死后和通过吲哚菁绿增强的 810nm 二极管激光脉络膜血管血栓形成实现脉络膜无灌注的动物中,确定了 NaFl 清除的动力学。用 ConA 对脉络膜扁平膜进行染色,验证了这些激光处理大鼠中的脉络膜无灌注。在体内,低剂量植入物在 25 分钟内释放了 >99%的药物,高剂量植入物在 60 分钟内释放了 >99%的药物。在 3 小时内,两种植入物剂量均>99%从巩膜组织中清除。到 7 小时时,低剂量植入物每毫克湿组织重量平均残留 0.14 +/- 0.131ng 的 NaFl,高剂量植入物每毫克湿组织重量残留 0.29 +/- 0.428ng 的 NaFl。相比之下,在 Tenon 下植入后 7 小时安乐死的动物中,低剂量植入物的巩膜组织中有 432.0 +/- 181.40ng 的 NaFl,高剂量植入物的巩膜组织中有 787.8 +/- 409.89ng 的 NaFl。在脉络膜血管选择性血栓形成的活动物中,两种植入物剂量的动物中,巩膜组织中残留的药物量在所有时间点均无显著差异。总之,从可生物降解的 Tenon 下植入物中递送至巩膜外的 NaFl 清除速度很快。清除机制在安乐死后明显降低,表明消除是通过主动的生理机制发生的,而不是通过被动扩散清除 (CL(diff))(Pfister 等人,2003 年)。有趣的是,脉络膜似乎没有发挥突出作用,因为巩膜外 NaFl 的清除不受脉络膜血流消除的影响。需要进一步的工作来描绘巩膜外药物清除的途径。