Sodium dodecyl sulfate and C31G as microbicidal alternatives to nonoxynol 9: Comparative sensitivity of primary human vaginal keratinocytes
Sodium dodecyl sulfate and C31G as microbicidal alternatives to nonoxynol 9: Comparative sensitivity of primary human vaginal keratinocytes
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DOI：
10.1128/aac.44.7.1954-1960.2000
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发表时间：
2000-07-01
影响因子：
4.9
通讯作者：
Wigdahl, B
中科院分区：
文献类型：
--
作者：
Krebs, FC;Miller, SR;Wigdahl, B
广谱阴道杀微生物剂必须对多种性传播疾病病原体有效，并且对阴道上皮内发现的细胞类型（包括阴道角质形成细胞）的毒性最低。我们评估了原代人阴道角质形成细胞对潜在的局部阴道杀微生物剂壬苯醇醚-9（N-9）、C31 G和十二烷基硫酸钠（SDS）的敏感性。直接免疫荧光和荧光激活细胞分选分析表明，原代阴道角质形成细胞表达上皮细胞特异性角蛋白。在连续48小时暴露期间比较阴道角质形成细胞对每种药剂的敏感性的实验表明，原代阴道角质形成细胞对N-9的敏感性几乎是对C31 G或SDS的5倍。为了评估多种杀微生物剂暴露对细胞活力的影响，将原代阴道角质形成细胞在78小时内暴露于N-9、C31 G或SDS三次。在这些实验中，在测试范围内的较低浓度下，细胞对C31 G比对N-9或SDS敏感得多。当选择试剂浓度以在三次每日暴露后产生25%活力的终点时，每次暴露以相同的恒定速率降低细胞活力。当检查连续48小时暴露期间的时间依赖性灵敏度时，暴露于C31 G 18小时导致细胞活力损失，直到至少24至48小时才由N-9或SDS引起。累积而言，这些结果揭示了体外培养的原代人阴道角质形成细胞群体中对N-9、C31 G或SDS的时间和浓度依赖性敏感性的重要变化。这些研究代表了阴道微环境体外建模和影响阴道局部杀微生物剂体内效力的因素研究的初步步骤。
A broad-spectrum vaginal microbicide must be effective against a variety of sexually transmitted disease pathogens and be minimally toxic to the cell types found within the vaginal epithelium, including vaginal keratinocytes. We assessed the sensitivity of primary human vaginal keratinocytes to potential topical vaginal microbicides nonoxynol-9 (N-9), C31G, and sodium dodecyl sulfate (SDS). Direct immunofluorescence and fluorescence-activated cell sorting analyses demonstrated that primary vaginal keratinocytes expressed epithelial cell-specific keratin proteins. Experiments that compared vaginal keratinocyte sensitivity to each agent during a continuous, 48-h exposure demonstrated that primary vaginal keratinocytes were almost five times more sensitive to N-9 than to either C31G or SDS. To evaluate the effect of multiple microbicide exposures on cell viability, primary vaginal keratinocytes were exposed to N-9, C31G, or SDS three times during a 78-h period. In these experiments, cells were considerably more sensitive to C31G than to N-9 or SDS at lower concentrations within the range tested. When agent concentrations were chosen to result in an endpoint of 25% viability after three daily exposures, each exposure decreased cell viability at the same constant rate. When time-dependent sensitivity during a continuous 48-h exposure was examined, exposure to C31G for 18 h resulted in losses in cell viability not caused by either N-9 or SDS until at least 24 to 48 h. Cumulatively, these results reveal important variations in time- and concentration-dependent sensitivity to N-9, C31G, or SDS within populations of primary human vaginal keratinocytes cultured in vitro. These investigations represent initial steps toward both in vitro modeling of the vaginal microenvironment and studies of factors that impact the in vivo efficacy of vaginal topical microbicides.