Department of Ophthalmology, Semmelweis University, Budapest, Hungary.
Invest Ophthalmol Vis Sci. 2010 Feb;51(2):1024-7. doi: 10.1167/iovs.09-4254. Epub 2009 Sep 24.
The aim of this study was to develop a model to investigate the permeability of the amniotic membrane (AM) to ofloxacin eye drops, a widely used topical antibiotic in ocular surface disease after AM transplantation.
AM pieces on cellulose acetate filter membranes were mounted in a vertical Franz-diffusion cell system equipped with an autosampler. In vitro release of 300 mg of 3% commercially available ofloxacin ophthalmic solution was determined by quantitative absorbance measurement carried out with a UV spectrophotometer (wavelength, 287 nm). Freshly prepared and cryopreserved AMs were compared. Filter membranes without AM served as positive controls.
Ofloxacin was detectable in the acceptor phase 1 minute after instillation, and a gradual increase of concentration could be detected in a period of 90 minutes in all groups. At 30 minutes 3.35% +/- 2.23% of ofloxacin penetrated the freshly prepared AM, 4.35% +/- 1.8% through cryopreserved AM compared with 17.52% +/- 3.91% filter membrane alone. At 90 minutes, penetration rates of ofloxacin were 5.04% +/- 1.11%, 5.26% +/- 3.21%, and 27.91% +/- 3.05%, respectively. Difference (P > 0.05; t-test) was not significant between freshly prepared and cryopreserved AMs. Compared with control, both membranes showed significant differences (P < 0.05; t-test) at all time points.
The in vitro model of the Franz-diffusion cell system was found to be applicable for drug permeability studies of human amniotic membranes to water-based solutions. The filter membrane and AM were permeable to a water-based solution of ofloxacin. Significant barrier function of the AM could be measured in ofloxacin permeability. Cryopreservation did not influence the permeability of the AM.
本研究旨在建立一个模型，以研究眼用抗生素氧氟沙星滴眼剂在羊膜（AM）移植后透过 AM 的渗透情况。
将纤维素醋酸酯滤膜上的 AM 片安装在配备自动进样器的垂直 Franz 扩散细胞系统中。通过定量吸光度测量法（波长 287nm），使用紫外分光光度计测定 300mg 市售 3%氧氟沙星滴眼液的体外释放情况。比较了新鲜制备和冷冻保存的 AM。无 AM 的滤膜作为阳性对照。
滴注后 1 分钟即可在接受相中检测到氧氟沙星，所有组在 90 分钟内均可检测到浓度逐渐增加。在 30 分钟时，新鲜制备的 AM 中氧氟沙星的穿透率为 3.35% +/- 2.23%，冷冻保存的 AM 为 4.35% +/- 1.8%，而单独的滤膜为 17.52% +/- 3.91%。90 分钟时，氧氟沙星的渗透速率分别为 5.04% +/- 1.11%、5.26% +/- 3.21%和 27.91% +/- 3.05%。新鲜制备和冷冻保存的 AM 之间的差异（P > 0.05；t 检验）不显著。与对照组相比，所有时间点滤膜和 AM 均有显著差异（P < 0.05；t 检验）。
Franz 扩散细胞系统的体外模型适用于水基溶液中人类 AM 对药物渗透的研究。滤膜和 AM 对水基氧氟沙星溶液具有渗透性。在氧氟沙星渗透中可以测量到 AM 的显著屏障功能。冷冻保存不会影响 AM 的通透性。
