426. IRRITATION TESTING OF CONTRACEPTIVE AND FEMININE-CARE PRODUCTS USING EPIVAGINAL, AN IN VITRO HUMAN VAGINAL-ECTOCERVICAL TISSUE MODEL.

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Ayehunie, S., Hayden, P., Cannon, C., Lamore, S., Klausner, M., Sheasgreen, J. MatTek Corporation, Ashland, MA. Presented at EUROTOX 2006, Cavtat and Dubrovnik, Croatia, September 20-24, (2006).

Summary:
This study by scientists at MatTek Corp. demonstrated that MatTek’s EpiVaginal human cervico-vaginal tissue equivalent is a useful, highly reproducible non-animal tool that can be used to assess the irritation potential of feminine hygiene products, as well enabling HIV infection, microbicides and drug absorption studies.

Normal human vaginal-ectocervical (VEC) cells were cultured to reconstruct VEC tissues in vitro (designated as EpiVaginal VEC-100). In some cultures, normal human dendritic cells were also incorporated (designated as EpiVaginal VLC-100-FT).

Both EpiVaginal™ tissues mimic native in vivo tissue in that they have basal, parabasal, glycogenated intermediate, and the superficial cell layers.

To test the utility of EpiVaginal VEC tissues, contraceptives, microbicides, anti-itch agents, and other vaginal care products were topically applied.

To mimic heterosexual HIV infection, the EpiVaginal VLC tissues were topically exposed to HIV-1 viruses.

Quality control (QC) testing on each batch of EpiVaginal tissue utilized Triton X-100 (1%) and water as positive and negative controls, respectively. The MTT assay was used to determine the exposure time necessary to decrease the EpiVaginal tissue viability to 50% (ET-50) for the positive control and 18 vaginal care products.

QC testing showed the EpiVaginal tissues to be highly reproducible; the average intra-lot coefficient of variation was <10% and ET-50s averaged 1.34 hr ± 0.21 (n=41 lots).

The EpiVaginal VEC tissue model discriminated between the mildness of vaginal care products. The ET-50 values ranged between 1.7-2.7 hr for feminine washes, 3.5-7.0 hr for contraceptives, 6.9->18 hr for anti-itch creams, and were > 18 hr for douches, lubricants, and anti-fungal creams. Released cytokines and gene expression levels showed that IL-1á, IL-1â, and IL-8 were associated with toxicity of vaginal care products.

Finally, EpiVaginal VLC tissues were infectible with macrophage-tropic and T-cell tropic HIV-1 strains as evidenced by DNA-PCR.

Based on these results, it is likely that the EpiVaginal VEC tissue model will serve as a useful, highly reproducible, non-animal tool to assess the irritation of vaginal care products. In addition, the EpiVaginal VLC tissue will enable studies pertaining to HIV infection, microbicides and drug absorption.

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EpiVaginal Data Sheet

EpiVaginal Specifications

EpiVaginal Technical References

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Applications: Cytokine expression/analysis, HIV/AIDS, Immunological research, Infectious disease research, Irritation - vaginal, Microbicides, Personal care products

Keywords: Anti-fungal creams, Contraceptives, Cytokines, DNA-PCR, Douches, EpiVaginal, Estrogen receptor, HIV-1, IL-1a, IL-1b, IL-8, Irritation, MTT, Microbicide, Microbicides, VEC DNA-PCR, Vaginal care products

Endpoints: ET-50, HIV infection, Histology (H & E staining), MTT viability

Materials Tested: Ammonium lauryl sulfate, Benzocaine, Clotrimazole, Glycerin, Miconazole nitrate, Nonoxynol-9 (N9), Povidene-iodine, Pramoxine HCl, Resorcinol, Tioconazole, Triton X-100, Vinegar

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