title: A study of the potential of the pig as a model for the vaginal irritancy of benzalkonium chloride in comparison to the nonirritant microbicide PHI-443 and the spermicide vanadocene dithiocarbamate.
abstract: A porcine model was established to test the mucosal toxicity potential of a thiophene thiourea (PHI-443)-based anti-HIV microbicide and a vanadocene-based spermicide, vanadocene dithiocarbamate (VDDTC) in comparison to benzalkonium chloride (BZK). Nine domestic pigs (Duroc) in nonestrus stage received a single intravaginal application of 2% BZK, 2% PHI-443, or 0.1% VDDTC-containing gel. At various times after gel application, cell differentials and levels of inflammatory cytokines (IL-1beta, IL-4, IL-6, IL-8, IL-10, IL-18, IFN-gamma, and TNF-alpha) in cervicovaginal lavage (CVL) fluid were monitored by flow cytometry and ELISA, respectively. Eight pigs were exposed intravaginally to a gel with and without BZK or VDDTC for 4 consecutive days and vaginal tissues were scored histologically for inflammation using a new scoring system. Only CVL fluid from pigs exposed to BZK showed a significant increase of IL-1beta, IL-8, and also IL-18 production when compared to the controls, PHI-443 or VDDTC-treated groups. Maximum levels of BZK-induced IL-1beta (100-fold), IL-8 (2,500-fold), IL-18 (80-fold), and IFN-gamma (10-fold) were found at 24 hours. In the in vivo porcine vaginal irritation model, increased levels of vaginal IL-1beta, IL-8, and IL-18 were associated with histological changes consistent with vaginal inflammation. These results demonstrate that key cervicovaginal inflammatory cytokines are useful in vivo biomarkers for predicting the mucosal toxicity potential of vaginal products in the physiologically relevant and sensitive porcine model.
authors.affiliation: Drug Discovery Program, Experimental Pathology, Parker Hughes Institute LLC, 2657 Patton Road, St. Paul, MN 55113, USA. odcruz@ih.org
authors.forename: Osmond J
authors.initials: OJ
authors.lastname: D'Cruz
authors.forename: Douglas
authors.initials: D
authors.lastname: Erbeck
authors.forename: Fatih M
authors.initials: FM
authors.lastname: Uckun
mesh_terms: D000818:Animals; D000891:Anti-Infective Agents, Local / Q000145:classification / Q000633:toxicity*; D001548:Benzalkonium Compounds / Q000145:classification / Q000633:toxicity*; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D007509:Irritants / Q000145:classification / Q000633:toxicity*; D023421:Models, Animal; D011725:Pyridines / Q000145:classification / Q000633:toxicity*; D023422:Skin Irritancy Tests / Q000379:methods; D013089:Spermatocidal Agents / Q000145:classification / Q000633:toxicity*; D013552:Swine; D007507:Therapeutic Irrigation; D013890:Thiourea / Q000031:analogs & derivatives* / Q000145:classification / Q000633:toxicity; D014621:Vagina / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D017968:Vanadium Compounds / Q000145:classification / Q000633:toxicity*
chemical_list: D000891:Anti-Infective Agents, Local; D001548:Benzalkonium Compounds; D016207:Cytokines; C403248:HI 443; D007509:Irritants; D011725:Pyridines; D013089:Spermatocidal Agents; D017968:Vanadium Compounds; C441979:bis(cyclopentadienyl)-N,N-diethyldithiocarbamato triflate salt; D013890:Thiourea