325. CHARACTERIZATION OF NEW BUCCAL AND GINGIVAL EPITHELIAL TISSUE MODELS.

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Kubilus¹, J., Breyfogle¹, B., Sheasgreen¹, J., Hayden¹, P., and Wertz², P., Dale³, B., Kimball³, J., and Klausner¹, M. ¹MatTek Corporation, Ashland, MA, ²University of Iowa, Iowa City, IA, ³University of Washington, Seattle, WA. J. Invest. Dermatol., 122, (3), A79, Abstract #474, (2004).

Summary: Three-dimensional models of the human oral epithelia, exhibiting a buccal or gingival phenotype have been developed using normal human oral epithelial cells cultured in serum free medium. Two versions of buccal tissue contain 8-12 and 25-35 cell layers (designated B12 and B35, respectively) with cells becoming increasingly squamous toward the apical surface have been developed. No evidence of cornification is present in histological slides and immuno-staining shows the expression of cytokeratin K13 in the suprabasal layers. Cytokeratin K4, the expression partner of K13, has also been detected by immuno-blotting. These features are characteristic of buccal epithelium. The gingival tissue (G13) has 9-13 layers of viable, nucleated cells and is partially cornified at the apical surface. When exposed to the surfactant Triton X-100, an exposure time of 52+/-20 minutes (n=31) reduces the viability of B12 to 50% as determined by an MTT assay. For G13 and B35, a similar exposure of >8 hours is required to damage the tissue to the same extent. Lipid analysis of B12 revealed that, of the ceramides important in the barrier of epidermis, only ceramide 2 (C2) was present, a result which matches that of human buccal tissue. B35 contained ceramides C2 and C3 in a ratio of 3.1:1 and G13 showed the presence of the three least polar ceramides, C1, C2, C3, in a ratio of 1: 8.2 : 4.5, respectively. The absolute amounts of ceramide in B12, B35, and G13 also varied significantly. Cultures contained 1.0, 4.5, and 8.2 ug/cm2, respectively. However, G13 was not as cornified as human hard palate, which contains all ceramides normally present in epidermis, C1-C7. The good correspondence between these tissues and those of native oral epithelia leads us to believe the tissues will be useful for a broad variety of basic and applied oral cavity studies.

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EpiOral/EpiGingival Data Sheet

EpiOral/EpiGingival Specifications

EpiOral/EpiGingival Technical References

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Applications: Characterization study, Irritation - oral

Keywords: Acyl glucosylated ceramide (AGC), Buccal, Buccal epithelium, Ceramide (CER AH), Ceramide 1 (CER EOS), Ceramide 2 (CER NS), Ceramide 3 (CER EOHP/NP), Ceramide 4 (CER AS), Ceramide 5 (CER NH), Ceramide 6 (CER AP), Ceramides, Cytokeratin K13, Cytokeratin K4, EVOM Epithelial Voltohmmeter, EpiGingival, EpiOral, GIN-100, Gingival, Gingival tissue, Gingivitis, Glucosphingolipid (GSL), Grafts, Gum disease, Lipid analysis, MTT, MTT ET-50 tissue viability assay, MTT assay, ORL-100, Oral cancer, Oral epithelia, Oral mucosal, Periodontal surgery, Phospholipids (PL), Three-dimensional differentiated tissues

Endpoints: Immunohistochemistry, Lipid analysis – high performance thin layer chromatography (HPTLC), MTT assay, MTT tissue viability assay, Transepithelial electrical resistance (TEER)

Materials Tested: Triton X-100

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