The Effects of Partial Demineralization and Fibronectin on Migration and Growth of Gingival Epithelial Cells on Cementum in vitro

Abstract

The capacity of mineralized cementum to support epithelial cell migration and growth and the effect that fibronectin and partial demineralization of cementum have on these processes were assessed in vitro. Dog gingival explants, 1 x 2 mm, were cultured on the cementum surfaces of pig root pieces in a defined medium consisting of DMEM and F₁₂ (1V/1V, transferrin, insulin, epidermal growth factor, cortisone, selenium, and high-density lipoprotein. Sixty root pieces were divided into four equal groups according to the treatment: (1) untreated mineralized cementum; (2) treated with 5 μg of fibronectin; (3) partially demineralized in 18% EDTA for 30 min; and (4) both partially demineralized and fibronectin-treated as above. Epithelial cell migration and growth on each of the four differently treated cementum surfaces were assessed histomorphometrically by means of scanning electron microscopy. The defined culture medium supported the selective migration and growth of epithelial cells from the gingival explants onto the mineralized cementum. This was confirmed by the positive immunostaining of these cells with antikeratin antibodies. Partial demineralization of cementum inhibited epithelial migration and growth by 83% and 91%, respectively. Fibronectin treatment did not affect epithelial cell migration and growth on mineralized cementum, but it decreased the degree of epithelial cell migration and growth inhibition on partially demineralized cementum to 57% and 43%, respectively. The results indicate that: (i) mineralized cementum may consist of components that are recognized by gingival epithelial cells and support their migration and growth in vitro; (ii) these components can be removed by demineralization; and (iii) fibronectin partially restores epithelial cell migration and growth on partial demineralized cementum in vitro.

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References

Boyko, G.A. ; Brunette, D.M. ; and Melcher, A.H. (1980): Cell Attachment to Demineralized Root Surfaces in vitro, J Periodont Res 15:297-303.

Caffesse, R.G. ; Holden, M.J. ; Kon, S. ; and Nasjleti, C.E. (1985): The Effect of Citric Acid and Fibronectin Application on Healing Following Surgical Treatment of Naturally Occurring Periodontal Disease in Beagle Dogs, J Clin Periodontol 12:578-590.

Donaldson, D.J. and Mahan, J.T. (1983): Fibronogen and Fibronectin as Substrates for Epidermal Cell Migration During Wound Closure, J Cell Sci 62:117-127.

Donaldson, D.J. ; Mahan, J.T. ; Hasty, D.L. ; McCarthy, J.B. ; and Furcht, L.T. (1985): Location of Fibronectin Domain Involved in Newt Epidermal Cell Migration, J Cell Biol 101:73-78.

Fernyhough, W.S. and Page, R.C. (1983): Attachment Growth and Synthesis by Human Gingival Fibroblasts on Demineralized or Fibronectin-treated Normal and Diseased Root Surface, J Periodontol 54:135-139.

Gottlow, J. ; Nyman, S. ; Karring, S. ; and Lindhe, J. (1986): Treatment of Localized Gingival Recessions with Coronally Displaced Flaps and Citric Acid, J Clin Periodontol 13:57-63.

Grinnell, F. (1984): Fibronectin and Wound Healing, J Cell Biochem 26:107-116.

Hynes, R.O. (1981): Fibronectin and Its Relation to Cellular Structure and Behaviour. In: Cell Biology of Extracellular Matrix, E.D. Hay , Ed., New York: Plenum Press.

Isidor, F. ; Karring, S. ; Nyman, S. ; and Lindhe, J. (1985): New Attachment Formation on Citric Acid Treated Roots, J Periodont Res 20:421-430.

10.

Karp, W. ; Sodek, J. ; Aubin, J.E. ; and Melcher, A.H. (1986): A Comparison of Fibronectin and Laminin Binding to Undemineralized and Demineralized Tooth Root Surfaces, J Periodont Res 21:30-38.

11.

Kashani, H.G. ; Magner, A.W. ; and Stahl, S.S. (1984): The Effect of Root Planing and Citric Acid Applications on Flap Healing in Humans, J Periodontol 55:679-683.

12.

Mosher, D.F. and McKeown-Longo (1985): Assembly of Fibronectin-containing Extracellular Matrix: A Glimpse of the Machinery, Biopolymers 24:199-210.

13.

Pearson, B.S. ; Klebe, R.J. ; Boyan, B.D. ; and Mosko-Wicz, D. (1988): Comments on the Clinical Application of Fibronectin in Dentistry, J Dent Res 67:515-517.

14.

Pitaru, S. ; Gray, A. ; Aubin, J.E. ; and Melcher, A.H. (1984): The Influence of the Morphological and Chemical Nature of Dental Surfaces on the Migration, Attachment and Orientation of Human Gingival Fibroblasts in vitro, J Periodont Res 19:408-418.

15.

Pitaru, S. ; Heckmati, H. ; Soldinger, M. ; and Savion, N. (1988): Growth and Migration of Gingival Epithelial Cells on Demineralized and Partially Demineralized Root Surfaces in an in vitro System, J Periodontol 59:531-534.

16.

Pitaru, S. and Melcher, A.H. (1983): Orientation of Gingival Fibroblasts and Newly-Synthesized Collagen Fibres in vitro: Resemblance to Transseptal and Dentogingival Fibres , J Periodont Res 18:483-500.

17.

Polson, A.M. and Proye, M.P. (1983): Fibrin Linkage: A Precursor for New Attachment , J Periodontol 54:141-147.

18.

19.

Ririe, G.M. ; Crigger, M. ; and Selvig, K.A. (1980): Healing of Periodontal Connective Tissues Following Surgical Wounding and Application of Citric Acid in Dogs, J Periodont Res 15:314-327.

20.

Salonen, J. and Santi, R. (1983): An Attempt to Simulate Junctional Epithelium of Human Gingiva in vitro, J Periodont Res 18:311-317.

21.

Takashima, A. and Grinnell, G. (1984): Human Keratinocyte Adhesion and Phagocytosis Promoted by Fibronectin, J Invest Dermatol 83:352-358.

22.

Takashima, A. and Grinnell, G. (1985): Fibronectin-mediated Keratinocyte Migration and Initiation of Fibronectin Receptor Function in vitro, J Invest Dermatol 85:304-308.

23.

Terranova, V.P. ; Francetti, L.C. ; Hic, S. ; and Wikesjo, U.M.E. (1987): Biochemically Mediated Periodontal Regeneration , J Periodont Res 22:248-251.

24.

Wirthlin, M.R. (1981): The Status of New Attachment Therapy, J Periodontol 52:529-544.

25.

Yamada, K.M. (1981): Fibronectin and Other Structural Proteins. In: Cell Biology of Extracellular Matrix, E.D. Hay , Ed., New York: Plenum Press .

26.

Yamada, K.M. ; Hayashi, M. ; Hirano, H. ; and Akiyama, S.K. (1984): Fibronectin and Cell Surface Interactions. In: The Role of Extracellular Matrix in Development, R.L. Trelstad , Ed., New York: Alan R. Liss, Inc.