Journal
TOXICOLOGY IN VITRO
Volume 20, Issue 5, Pages 689-698Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.tiv.2005.10.002
Keywords
vagina; irritation; feminine care; tissue model; contraceptives; microbicides; toxicity; cytokines
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Funding
- PHS HHS [2R44A147792] Funding Source: Medline
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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 in vitro tissue model was reconstructed using normal VEC epithelial cells and is well stratified, containing 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 intra-lot 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 degrees C on agarose gels. The utility of the 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 2.7 h for feminine washes, 3.9-6.7 h for spermicides, 6.8-18 h for anti-itch creams, and > 18 It for douches, lubricants, and anti-fungal creams. Studies of cytokines released from VEC cultures following product application showed that elevated concentrations of IL-1 alpha and IL-1 beta were associated with toxicity of test materials. In conclusion, the VEC tissue model is a highly reproducible, non-animal means to assess the irritation of contraceptives, microbicides, and vaginal-care products. (c) 2005 Elsevier Ltd. All rights reserved.
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