Organotypic raft cultures for the in vitro evaluation of vaginal microbicidal agents. Microbicides have epithelium-specific effects when repeatedly added onto organotypic human keratinocyte cultures.
OBJECTIVE
The authors tested for keratinocyte raft cultures to compare vaginal microbicides that could be used at high frequency to prevent transmission of sexually transmitted diseases but should have minimal effects on the vaginal epithelium.


METHODS
Effects of previously described microbicides were analyzed on normal foreskin keratinocytes and human papillomavirus-immortalized cervical epithelial cells in raft cultures after hematoxylin-eosin staining and immunostainings for keratins, filaggrin, proliferating cell nuclear antigen and Ki67-antigen; toxicity toward feeder cells was monitored by quantifying interleukin-6 release.


RESULTS
Repeated addition of microbicides into the medium resulted in histologic alterations at concentrations predictable from cytotoxicity tests performed in submerged cultures and correlated with reduced interleukin-6 release. Normal interleukin-6 release was observed when histologic alterations were induced by microbicides applied onto the rafts.


CONCLUSIONS
Adding microbicides onto raft cultures at effective virucidal concentrations shows harmful effects on epithelial tissues undetectable in submerged cultures.
Aebischer
F
F
Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98102, USA.
McDougall
J K
JK
D000282:Administration, Intravaginal; D000890:Anti-Infective Agents / Q000008:administration & dosage* / Q000009:adverse effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004848:Epithelium / Q000187:drug effects; D005260:Female; D000091344:Filaggrin Proteins; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D015603:Keratinocytes / Q000187:drug effects*; D012749:Sexually Transmitted Diseases / Q000517:prevention & control*; D014621:Vagina / Q000187:drug effects*
D000890:Anti-Infective Agents; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D015850:Interleukin-6