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Antimicrobial Agents and Chemotherapy, July 2000, p. 1954-1960, Vol. 44, No. 7
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Sodium Dodecyl Sulfate and C31G as Microbicidal Alternatives to Nonoxynol 9: Comparative Sensitivity of Primary Human Vaginal Keratinocytes

Fred C. Krebs,1 Shendra R. Miller,1 Bradley J. Catalone,1 Patricia A. Welsh,1 Daniel Malamud,2,3 Mary K. Howett,1 and Brian Wigdahl1,*

Department of Microbiology and Immunology, College of Medicine, The Pennsylvania State University, Hershey, Pennsylvania 17033,1 and Department of Biochemistry, School of Dental Medicine, University of Pennsylvania,2 and Biosyn, Inc.,3 Philadelphia, Pennsylvania 19104

Received 16 November 1999/Returned for modification 19 January 2000/Accepted 25 April 2000

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.

* Corresponding author. Mailing address: Department of Microbiology and Immunology (H107), The Pennsylvania State University, College of Medicine, 500 University Drive, P.O. Box 850, Hershey, PA 17033. Phone: (717) 531-8258. Fax: (717) 531-5580. E-mail: bwigdahl{at}psu.edu.

Antimicrobial Agents and Chemotherapy, July 2000, p. 1954-1960, Vol. 44, No. 7
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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