Department of Ophthalmology, Paulista School of Medicine, Federal University of Sao Paulo, UNIFESP, Sao Paulo, Brazil.
Invest Ophthalmol Vis Sci. 2011 Dec 9;52(13):9333-8. doi: 10.1167/iovs.11-8382.
To assess the Acanthamoeba trophozoite viability in vitro and treatment of Acanthamoeba keratitis in a hamster model using ultraviolet light A (UV-A) and riboflavin (B2).
A sample of Acanthamoeba sp. cultured was transferred to a 96-well plate and exposed to B2 and the UV-A light (365 nm wavelength) at a power density of 3 mW/cm(2), 8 mm spot diameter, for 30 minutes. The exposure was done in triplicate. Control groups were prepared in triplicate as well: blank control, UV-A only, riboflavin only, and dead control. Cell viability assessment was done using the trypan blue dye exclusion method. Acanthamoeba keratitis was induced in Chinese hamsters; who were randomly assigned to one of the animal groups: UV-A + B2, propamidine isethionate (Brolene; Sanofi-Aventis, Ellerslie, Auckland, Australia), UV-A + B2 + propamidine isethionate (Brolene), only UV-A, only B2, and blank. Throughout the 14 days after treatment the animals were examined clinically. Histology and clinical scores of all groups were compared.
The in vitro study showed no difference between the treatment group UV-A + B2 and the control groups. In the hamster keratitis model a significant improvement of clinical score was observed for the groups propamidine isethionate (Brolene) and UV-A + B2 + propamidine isethionate (Brolene) (P = 0.0067). Also a significant worsening of clinical score was observed in the other groups: UV-A + B2 group (P = 0.0084), only UV-A (P = 0.0078), B2 only (P = 0.0084), and blank (P = 0.0082). No difference was observed between propamidine isethionate (Brolene) and UV-A + B2 + propamidine isethionate (Brolene).
The adjunctive use of UV-A and B2 therapy did not demonstrate antitrophozoite activity; in vivo UV-A and B2 did not demonstrate efficacy in this model.
评估紫外线 A（UV-A）和核黄素（B2）对体外棘阿米巴滋养体活力的影响，并在仓鼠模型中治疗棘阿米巴角膜炎。
将培养的棘阿米巴样本转移到 96 孔板中，在功率密度为 3 mW/cm(2)、8mm 光斑直径的条件下，用 UV-A（365nm 波长）照射 B2，照射时间 30 分钟。每组重复 3 次。同样制备了 3 组对照：空白对照、仅 UV-A、仅核黄素和死亡对照。使用台盼蓝染料排除法评估细胞活力。在中华仓鼠中诱导棘阿米巴角膜炎；将其随机分配到以下动物组之一：UV-A+B2、丙脒腙（Brolene；赛诺菲-安万特，奥克兰，新西兰）、UV-A+B2+丙脒腙（Brolene）、仅 UV-A、仅 B2 和空白。在治疗后的 14 天内，对动物进行临床检查。比较所有组的组织学和临床评分。
体外研究表明，治疗组 UV-A+B2 与对照组之间无差异。在仓鼠角膜炎模型中，丙脒腙（Brolene）和 UV-A+B2+丙脒腙（Brolene）组的临床评分显著改善（P=0.0067）。其他组的临床评分也显著恶化：UV-A+B2 组（P=0.0084）、仅 UV-A 组（P=0.0078）、仅 B2 组（P=0.0084）和空白组（P=0.0082）。丙脒腙（Brolene）与 UV-A+B2+丙脒腙（Brolene）之间无差异。
辅助使用 UV-A 和 B2 治疗并未显示出抗滋养体活性；体内 UV-A 和 B2 在该模型中无效。
