ORGANOTYPIC HUMAN VAGINAL-ECTOCERVICAL TISSUE MODEL FOR IRRITATION STUDIES OF SPERMICIDES, MICROBICIDES, AND FEMININE-CARE PRODUCTS.

This study by scientists at Boston University School of Medicine and MatTek Corp. demonstrated that MatTek’s EpiVaginal in vitro human cervico-vaginal tissue equivalent is a highly reproducible, non-animal means to assess the irritation potential of topically applied contraceptives, microbicides and vaginal care products. EpiVaginal, a three-dimensional organotypic vaginal-ectocervical (VEC) tissue model, has been developed to test the irritation of topically applied spermicides, microbicides, and vaginal-care products. The EpiVaginal in vitro tissue model was reconstructed using normal VEC epithelial cells and is well stratified, contained differentiated basal, suprabasal, intermediate and superficial cell layers similar to in vivo tissue. The intermediate and superficial cell layers contain glycogen, and the expression of cytokeratins 13 and 14 in the tissue also parallels that of native tissue. The MTT viability assay and histological assessment were used to test inter-lot and intra-lot reproducibility. The MTT average intralot coefficient of variation (CV) was less than 10% and the time required to reduce tissue viability by 50% (ET-50) following application of 1% Triton X-100 averaged 1.25 ± 0.24 h (n=23) upon completion of the 11-day culture period and 1.30 ± 0.19 for the same tissues stored overnight at 4°C on agarose gels. The utility of the EpiVaginal VEC model for irritation studies was examined by testing commercially available products using the MTT assay and histological assessment. The average ET-50 values ranged between 1.8 and 2.7h for feminine washes, 3.9-6.7h for spermicides, 6.8-18 h for anti-itch creams, and >18 h for douches, lubricants, and anti-fungal creams. Studies of cytokines released from EpiVaginal tissue cultures following product application showed that elevated concentration of IL-1á and IL-1â were associated with toxicity of test materials. In conclusion, the EpiVaginal tissue model is a highly reproducible, non-animal means to assess the irritation of contraceptives, microbicides, and vaginal-care products.

Cytokines, ET-50, Feminine care products, High throughput screening, IL-1a, IL-1b, In vivo vaginal models, Initial screening, Inter-lot, Intra-lot reproducibility, Irritation, MTT, MTT tissue viability assay, Microbicide, Microbicides, Robotic-compatible, Spermicides, Toxicity, Triton X-100, VEC, Vaginal-ectocervical, Vaginal-ectocervical tissue

Anti-itch cream: benzocaine (20%), BAK (0.13%), Anti-itch cream: benzocaine (20%), resorcinol (3%), Anti-itch cream: benzocaine (5%), resorcinol (2%), Anti-itch cream: bramoxine HCl (1&), glycerin (39%), Antifungal cream: clotrimazole (2%), Antifungal cream: miconazole nitrate (2%), Antifungal cream: tioconazole (6.5%), Douche: povidene-iodine, Douche: vinegar, Feminine wash (normal): ammonium laureth sulfate, Feminine wash (sensitive): ammonium lauryl sulfate, Feminine wash: ammonium laureth sulfate, Personal lubricant, Spermicide: nonoxynol-9 (2%), Spermicide: nonoxynol-9 (3%), Spermicide: nonoxynol-9 (4%)