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HUMAN β-DEFENSIN EXPRESSION IN A NEW BUCCAL EPITHELIAL TISSUE MODEL.

Kubilus1, J., Breyfogle1, B., Sheasgreen1, J., Hayden1, P., Klausner1, M., Dale2, B.A., Kimball2, J.R. 1MatTek Corporation, Ashland, MA. 2University of Washington, Seattle, WA.
Abstract

Oral epithelia function as microbial barriers against bacteria and yeast that are resident in the oral cavity. These microbes can be categorized as non-pathogenic (commensal) and pathogenic (resulting in cavities, gingivitis, and periodontal disease, etc.). The epithelium is actively involved in recognition and response to bacteria, setting into motion the innate immune responses that are an important part of periodontal health and disease. One epithelial response is the expression of antimicrobial peptides of the human beta-defensin (hBD) family. A new, three-dimensional model of the human buccal epithelium, based on normal human oral epithelial cells cultured in serum free medium, has been developed. Histologically, the tissue is 20-30 cell layers thick with cells becoming increasingly squamous toward the apical surface. No evidence of cornification is present 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. Additional immuno-staining revealed the presence of hBD-1 and the recently discovered hBD-3 in the suprabasal layers; hBD2 was not expressed. The hBD-1 and hBD-2 results parallel those of un-inflamed epithelia, however the hBD-3 result has not been previously reported. Based on these initial results, further investigation of the tissue model’s hBD expression in unstimulated and bacterially challenged tissue is warranted. Note: Subsequent to submission of this abstract, additional data were obtained which have been included in this presentation.

Keywords

Anti-microbial, Anti-microbial activity, Anti-microbial peptide, Anti-microbial peptides, Antimicrobial, Antimicrobial activity, Antimicrobial peptide, Antimicrobial peptides, Bacterially challenged tissue, Buccal, Buccal epithelium, Cavities, Commensal, Cytokeratin K13, Cytokeratin K4, EpiOral, Epithelial response, Epithelium is actively, Gingivitis, HBD, HBD-1, HBD-3, Human beta-defensin, Human buccal epithelium, Human oral epithelial cells, Immuno-blotting, Immuno-staining, MTT, MTT ET-50 tissue viability assay, MTT assay, Microbial barrier, Non-pathogenic, Oral cavity, Oral epithelia, Pathogenic, Periodontal disease, Suprabasal layer

Materials Tested

SLS, Sodium lauryl sulfate, Toothpaste

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