Youn Hyun-Yi, Moran Kelley L, Oriowo Olanrewaju M, Bols Niels C, Sivak Jacob G
School of Optometry, University of Waterloo, Waterloo, ON, Canada N2L 3G1.
Toxicol In Vitro. 2004 Dec;18(6):841-52. doi: 10.1016/j.tiv.2004.04.007.
To evaluate in vitro methods for testing the toxicity of the surfactants, sodium dodecyl sulfate (SDS) and benzalkonium chloride (BAK), and Ultraviolet (UV)-B radiation to the bovine lens.
Lenses were dissected from bovine eyes--obtained from a local abattoir--and incubated in M199 culture medium at 37 degrees C, with 4% CO(2) and 96% air atmosphere. For the SDS and BAK experiments, the lenses (n = 153) were exposed directly to 0.001%, 0.01%, 0.1%, and 1.0% solutions for 15 min. These lenses were then rinsed five times each with saline and medium. Another group of lenses (n = 36) was irradiated with broadband UV-B at energy levels of 1.0 and 2.0 J/cm(2) (0.445 and 0.89 J/cm(2) in the biologically effective energy levels). For all of the above experiments, lens optical quality and cellular viability of lens epithelial cells were evaluated.
The analysis of optical quality, using a scanning laser in vitro assay system, of exposed lenses treated with SDS and BAK at concentrations of 0.01%, 0.1%, and 1.0%, and with UV-B at energy levels of 0.445 and 0.89 J/cm(2) showed a dose- and time-dependent increase in back vertex distance (BVD) variability, indicating loss of sharp focus in comparison with control lenses. Both 0.001% SDS and 0.001% BAK-treated lenses did not show any optical damage until 8-days after exposure. Lenses treated with 0.01% SDS showed recovery from optical damage 6-days later after exposure. Optical damage was not shown immediately for UV-B-exposed lenses. The Alamar Blue assay data for SDS, BAK and UV-B-exposed lenses, except the 0.001% SDS treated lens group, showed also dose- and time-dependent decreases in cellular viability in comparison with the control lenses, and there was no cellular recovery during the entire culture period. Lenses treated with 0.001% SDS did not show biological damage until 8-days after exposure. It appears that cellular changes appeared earlier than optical changes.
The findings suggest that cultured bovine lenses can be evaluated by assays that probe optical properties and cellular function after exposure to surfactants and UV-B irradiation, and that the optical and biological assay methods are valuable for in vitro mild ocular toxicity research.
评估检测表面活性剂十二烷基硫酸钠（SDS）和苯扎氯铵（BAK）以及紫外线（UV）-B辐射对牛晶状体毒性的体外方法。
从当地屠宰场获取的牛眼中摘取晶状体，在37℃、含4%二氧化碳和96%空气的M199培养基中培养。对于SDS和BAK实验，将晶状体（n = 153）直接暴露于0.001%、0.01%、0.1%和1.0%的溶液中15分钟。然后用生理盐水和培养基各冲洗这些晶状体五次。另一组晶状体（n = 36）用能量水平为1.0和2.0 J/cm²（生物有效能量水平为0.445和0.89 J/cm²）的宽带UV-B照射。对于上述所有实验，评估晶状体的光学质量和晶状体上皮细胞的细胞活力。
使用扫描激光体外检测系统对用0.01%、0.1%和1.0%的SDS和BAK以及能量水平为0.445和0.89 J/cm²的UV-B处理的暴露晶状体进行光学质量分析，结果显示后顶点距离（BVD）变异性呈剂量和时间依赖性增加，表明与对照晶状体相比焦点清晰度丧失。0.001% SDS和0.001% BAK处理的晶状体在暴露后8天内未显示任何光学损伤。用0.01% SDS处理的晶状体在暴露6天后光学损伤有所恢复。UV-B照射的晶状体未立即显示光学损伤。除0.001% SDS处理的晶状体组外，SDS、BAK和UV-B照射的晶状体的阿拉玛蓝检测数据也显示与对照晶状体相比细胞活力呈剂量和时间依赖性下降，并且在整个培养期间没有细胞恢复。用0.001% SDS处理的晶状体在暴露后8天内未显示生物损伤。似乎细胞变化比光学变化出现得更早。
研究结果表明，培养的牛晶状体可通过检测暴露于表面活性剂和UV-B照射后的光学特性和细胞功能的方法进行评估，并且光学和生物学检测方法对于体外轻度眼毒性研究具有重要价值。
