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CHARACTERIZATION OF NEW BUCCAL AND GINGIVAL EPITHELIAL TISSUE MODELS.
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
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 |
