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Reconstituted human gingival epithelium: Nonsubmerged in vitro model

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Summary

Many studies have shown that human gingival keratinocytes grown in submerged culture fail to attain optimal differentiation. This study reports an in vitro culture system for oral gingival epithelial cells, in which they are grown at the air-liquid interface, on polycarbonate inserts, in the presence of an NIH-3T3 feeder layer. This model was compared with two submerged culture methods for gingival keratinocytes, on type I collagen gel and on an NIH-3T3 feeder layer. Transmission electron microscopy showed an advanced level of stratification (over six layers of cells) for cultures grown at the air-liquid interface. Immunofluorescence and electrophoretic patterns showed the presence of cytokeratins 10 and 11 in cytoskeletal protein extracts of these cultured keratinocytes. In this air-liquid interface culture model, in the presence of NIH-3T3 feeder cells, keratinocytes can achieve an advanced level of stratification and differentiation and a resemblance to in vivo gingiva. The obtention of a highly differentiated epithelium will permit in vitro pharmacological studies and studies on the biocompatability of certain alloys with the superficial periodontium; it will also provide grafts for patients undergoing periodontal surgery.

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References

  1. Achtstaetter, T.; Hatzfeld, M.; Quinlan, R. A., et al. Separation of cytokeratin polypeptides by gel electrophoretic and chromatographic techniques and their identification by immunoblotting. Methods Enzymol. 134:355–371; 1986.

    Article  PubMed  CAS  Google Scholar 

  2. Alitalo, K.; Kuismanen, E.; Myllylä, R., et al. Extracellular matrix proteins of human epidermal keratinocytes and feeder 3T3 cells. J. Cell Biol. 94:497–505; 1982.

    Article  PubMed  CAS  Google Scholar 

  3. Ansorge, W. Fast and sensitive detection of protein and DNA bands by treatment with potassium permanganate. J. Biochem. Biophys. Methods 11:13–20; 1985.

    Article  PubMed  CAS  Google Scholar 

  4. Asselineau, D.; Bernhard, B.; Bailly, C., et al. Epidermal morphogenesis and induction of the 67 kD keratin polypeptide by culture of human keratinocytes at the liquid-air interface. Exp. Cell Res. 159:536–539; 1985.

    Article  PubMed  CAS  Google Scholar 

  5. Bell, E.; Sher, S.; Hull, B., et al. The reconstitution of living skin. J. Invest. Dermatol. 81:2–10; 1983.

    Article  Google Scholar 

  6. Boyce, S. T.; Ham, R. G. Calcium regulated differentiation of neural human epidermal keratinocytes defined clonal culture and a serum-free serial cultures. J. Invest. Dermatol. 81:33–40; 1983.

    Article  Google Scholar 

  7. Calle, A.; Poly, H.; Faure, M., et al. Ultrastructure of multilayered cultures of human gingival epithelial cells: attachment to enamel surfaces in vitro. Biol. Cell. 58:95–98; 1986.

    PubMed  CAS  Google Scholar 

  8. Cooper, D.; Schermer, A.; Sun, T. T. Classification of human epithelia and their neoplasms using monoclonal antibodies to keratins: strategies, applications, and limitations. Lab. Invest. 52:243; 1985.

    PubMed  CAS  Google Scholar 

  9. Franke, W. W.; Schiller, D. L.; Moll, R., et al. Diversity of cytokeratins. Differentiation specific expression of cytokeratin polypeptides in epithelial cells and tissues. J. Mol. Biol. 153:933–959; 1981.

    Article  PubMed  CAS  Google Scholar 

  10. Goldberg, M. Tissus non minéralisés et milieu buccal. Paris: Masson; 1993.

    Google Scholar 

  11. Gosselin, F.; Gervaise, M.; Neveux, Y., et al. Gingiva-equivalent: the reconstitution in vitro of human gingiva. J. Dent. Res. 69:188; 1990.

    Google Scholar 

  12. Gosselin, F.; Magloire, H.; Joffre, A., et al. Cytokeratins as molecular markers in the evaluation of precise differentiation stage of human gingival epithelium reconstituted in vitro. Arch. Oral Biol. 35:217S-221S; 1990.

    Article  PubMed  CAS  Google Scholar 

  13. Katz, A. B.; Taichman, L. H. Epidermis as a secretory tissue: an in vitro tissue model to study keratinocyte secretion. J. Invest. Dermatol. 102:55–60; 1994.

    Article  PubMed  CAS  Google Scholar 

  14. Korge, B.; Stadler, R.; Mischke, D. Effect of retinoids on hyperproliferation-associated keratins K6 and K16 in cultured human keratinocytes: a quantitative analysis. J. Invest. Dermatol. 95:450–455; 1990.

    Article  PubMed  CAS  Google Scholar 

  15. Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685; 1970.

    Article  PubMed  CAS  Google Scholar 

  16. Moll, R.; Franke, W. W.; Schiller, D. L. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell 31:11–24; 1984.

    Article  Google Scholar 

  17. Muller-Glauser, W.; Preisig, E. The effect of cholera toxin and epidermal growth factor on the in vitro growth of human oral epithelial cells. Arch. Oral Biol. 28:765–769; 1983.

    Article  PubMed  CAS  Google Scholar 

  18. Oda, D.; Dale, B. A.; Bourekis, G. Human oral epithelial cell culture II. Keratin expression in fetal and adult gingival cells. In Vitro Cell. Dev. Biol. 26:596–603; 1990.

    Article  PubMed  CAS  Google Scholar 

  19. Oda, D.; Watson, E. Human oral epithelial cell culture I. Improved conditions for reproducible culture in serum-free medium. In Vitro Cell. Dev. Biol. 26:589–595; 1990.

    Article  PubMed  CAS  Google Scholar 

  20. O’Farrell, P. H. High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250:4007–4021; 1975.

    PubMed  CAS  Google Scholar 

  21. O’Farrell, P. Z.; Howard, M.; O’Farrel, P. H. High resolution two dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142; 1977.

    Article  PubMed  CAS  Google Scholar 

  22. Ouhayoun, J. P.; Gosselin, F.; Forest, N., et al. Cytokeratin patterns of human oral epithelia: differences in cytokeratin synthesis in gingival epithelium and the adjacent alveolar mucosa. Differenciation 30:123–129; 1985.

    Article  CAS  Google Scholar 

  23. Pan, Y. M.; Firth, J. D.; Salonen, J. I., et al. Multilayer culture of periodontal ligament epithelial cells: a model for junctional epithelium. J. Periodontal Res. 30:97–107; 1995.

    Article  PubMed  CAS  Google Scholar 

  24. Poly, H.; Couble, M. C.; Hartmann, D. J., et al. Electron immunolocalization of type IV collagen, laminin and fibronectin synthesized by multilayered cell cultures from human oral epithelium. J. Periodontal Res. 23:252–257; 1988.

    Article  PubMed  CAS  Google Scholar 

  25. Pruniéras, M.; Régnier, M.; Woodley, D. Methods for cultivation of keratinocytes with an air-liquid interface. J. Invest. Dermatol. 81:28–33; 1983.

    Article  Google Scholar 

  26. Rheinwald, J. G.; Green, H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6:331–334; 1975.

    Article  PubMed  CAS  Google Scholar 

  27. Sawaf, M. H.; Ouhayoun, J. P.; Forest, N. Cytokeratin profiles in oral epithelia: a review and a new classification. J. Biol. Buccale 19:187–198; 1991. The publication of this journal has been interrupted in 1992.

    PubMed  CAS  Google Scholar 

  28. Schroeder, H. E. Oral structural biology: embryology, structure, and function of normal hard and soft tissues of the oral cavity and temporomandibular joints. New York: Thieme Medical; 1991.

    Google Scholar 

  29. Shabana, A. H. M.; Khattab, R.; Sawaf, M. H., et al. A switch in cytokeratin expression and intermediate filament organization associated with epithelial stratification. J. Biol. Buccale 20:33–43; 1992.

    PubMed  CAS  Google Scholar 

  30. Southgate, J.; Williams, H. K.; Trejdosiewicz, L. K., et al. Primary culture of human oral epithelial cells. Lab. Invest. 56:211–223; 1987.

    PubMed  CAS  Google Scholar 

  31. Staiano-Coico, L.; Helm, R. E.; McMahon, C. K., et al. Sodium-N-butyrate induces cytoskeletal rearrangements and formation of cornified envelopes in cultured adult human keratinocytes. Cell Tissue Kinet. 22:361–375; 1989.

    PubMed  CAS  Google Scholar 

  32. Terranova, V. P.; Lyall, R. M. Chemotaxis of human gingival epithelial cells to laminin. A mechanism for epithelial cell apical migration. J. Periodontol. 57:311–317; 1986.

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Equipe de Biologie Buccale, Université de Rennes, I, 2 place Pasteur, 35000, Rennes, France

    Agnes Delcourt-Huard & Martine Bonnaure-Mallet

  2. Unité INSERM 49 Hôpital Pontchaillou, France

    Anne Corlu

  3. Laboratoire d’Histophysiologie et Pathologie des tissus dentaires, Faculté d’Odontologie de Lyon, France

    Annick Joffre & Henry Magloire

Authors
  1. Agnes Delcourt-Huard
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  2. Anne Corlu
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  3. Annick Joffre
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  4. Henry Magloire
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  5. Martine Bonnaure-Mallet
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Delcourt-Huard, A., Corlu, A., Joffre, A. et al. Reconstituted human gingival epithelium: Nonsubmerged in vitro model. In Vitro Cell.Dev.Biol.-Animal 33, 30–36 (1997). https://doi.org/10.1007/s11626-997-0019-0

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  • DOI: https://doi.org/10.1007/s11626-997-0019-0

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