Kinetic Aspects of Gingival and Periodontal Ligament Fibroblast Attachment to Surface-conditioned Dentin

Abstract

An in vitro model has been developed for measurement of initial attachment of 3H-labeled human gingival fibroblasts (HGF) and periodontal ligament cells (HPDL cells) to sieved dentin powder. Reproducible attachment was obtained and was closely related to the number of plated cells, the amount of dentin powder, the temperature, and the time (from 0 to 6 h) of incubation. HGF and HPDL cells had a high affinity for untreated dentin. Heat-denatured BSA coating of the dentin fully inhibited cell attachment, while a fibronectin coating had no significant effect. A surface conditioning of the dentin grains by 3% citric acid or by 2.5% minocycline-HCl increased the initial attachment of HPDL cells significantly (p < 0.05). Minocycline-HCl was more efficient than citric acid in that respect. In addition, HGF attachment to untreated dentin was highly significantly improved (p < 0.005) by a pre-incubation of the cells with 50 μg/mL of minocycline, and there was also indication (p = 0.067) of improvement by the presence of minocycline in the attachment medium, with a maximum of efficacy at 110 μg/mL. These results suggest that minocycline-HCl directly influenced the attachment properties of fibroblastic cells and that citric acid and minocycline-HCl could act by different mechanisms. HGF and HPDL cells originating from the same patient displayed no significant difference in their attachment to dentin in this model.

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References

Becker W. , Becker BE , Prichard JF , Caffesse R. , Rosenberg E. , Gian-Grasso J. (1987). Root isolation for new attachment procedures. A surgical and suturing method: Three case reports. J Periodontol 58:819-826.

Bowers GM , ChadroffB. , Carnevale R. , Mellonig J. , Corio R. , Emerson J. , et al.(1989a). Histologic evaluation of new attachment apparatus formation in humans. Part I. J Periodontol 60:664-674. Bowers GM , Chadroff B. , Carnevale R. , Mellonig J. , Corio R. , Emerson J. , et al. (1989 b). Histologic evaluation of new attachment apparatus formation in humans. Part II. J Periodontol 60:675-693. Boyko GA , Brunette DM , Melcher AH (1980). Cell attachment to demineralized root surfaces in vitro. J Periodontol 15:297-303. Caffesse RG , Nasjleti CE , Anderson GB , Lopatin DE , Smith BA , Morrison EC (1991). Periodontal healing following guided tissue regeneration with citric acid and fibronectin application. J Periodontol 62:21-29.

Fernyhough W. , Page RC ( 1983). Attachment, growth and synthesis by human gingival fibroblasts on demineralized or fibronectin-treated normal and diseased tooth roots. J Periodontol 54 :133-140.

Garrett JS , Crigger M. , Egelberg J. (1978). Effects of citric acid on diseased root surfaces . J Periodont Res 13:155-163.

Hanes PJ , Polson AM (1989). Cell and fiber attachment to demineralized cementum from normal root surfaces. J Periodontol 60:188-198.

Hanes PJ , Polson AM , Frederick GT (1988). Initial wound healing attachments to demineralized dentin. J Periodontol 59:176-183. Hanes PJ , Polson AM , Ladenheim S. (1985). Cell and fiber attachment to demineralized dentin from normal root surfaces . J Periodontol 56:752-764.

Heritier M. (1982). Etude au microscope electronique a balayage et a transmission des effets d'une solution d'acide citrique sur la dentine radiculaire humaine. J Biol Buccale 10:147-167.

Karp W. , Sodek J. , Aubin JF , Melcher AH (1986). A comparison of fibronectin and laminin binding to undemineralized and demineralized tooth root surface. J Periodont Res 21:30-38. Kleinman HK , Klebe RJ , Martin GF (1981). Role of collagenous matrices in the adhesion and growth of cells. J Cell Biol 88 :473-485.

Lowenberg BF , Aubin JE , Deporter DA , Sodek J. , Melcher AH (1985). Attachment, migration and orientation of human gingival fibroblasts to collagen-coated, surface-demineralized, and untreated root slices. J Dent Res 64:1106-1110.

10.

Lowenberg BF , Aubin JE , Pitaru S. , Melcher AH (1984). A new chromium 51 assay for accurate measurement of cell attachment to demineralized and non-demineralized dentin in vitro . J Periodont Res 19:40-50.

11.

Lowenberg BF , Thibault J. , Lawrence C. , Sodek J. (1986). The influ

12.

ence of chemically-induced modifications of root surfaces on cell migration, attachment, and orientation. J Dent Res 65 :1010-1015.

13.

Lynch SE , Williams RC , Polson AM , Howell TH , Reddy MS , Zappa UE , et al. (1989). A combination of platelet-derived and insulin-like growth factors enhances periodontal regeneration. J Clin Periodontol 16:545-548.

14.

Mendieta C. , Caravana C. , Fine DH ( 1990). Sorption of fibronectin to human root surfaces in vitro . J Periodontol 61:254-260.

15.

Polson AM , Frederick GT (1985). Cell processes in dentin tubules during early phases of attachment to demineralized periodontitis-affected surfaces. J Clin Periodontol 12:162-169.

16.

Pontoriero R. , Lindhe J. , Nyman S. , Karring T. , Rosenberg E. , Sanavi F. (1988). Guided tissue regeneration in degree II furcation-involved mandibular molars: A clinical study. J Clin Periodontol 15:247-254.

17.

Pontoriero R. , Lindhe J. , Nyman S. , Karring T. , Rosenberg E. , Sanavi F. (1989). Guided tissue regeneration in the treatment of furcation defects in mandibular molars. J Clin Periodontol 16:170-174. Ragnarsson B. , Carr G. , Daniel JC (1985). Isolation and growth of human periodontal ligament cells in vitro. J Dent Res 64 :1026-1030.

18.

Register AA , Burdick FA (1975). Accelerated reattachment with cementogenesis to dentin, demineralized in situ. I. Optimum range. J Periodontol 46:646-655.

19.

Somerman MJ , Archer SY , Imm GR , Foster RA (1988a). A comparative study of human periodontal ligament cells and gingival

20.

fibroblasts in vitro. J Dent Res 67:66-70.

21.

Somerman MJ , Foster RA , Progebin K. , Wynn RL ( 1988b). Effects of minocycline on fibroblasts attachment and spreading. JPeriodont Res 23:154-159.

22.

Terranova VP , Franzetti LC , Hic S. , Di Florio RM , Lyall RM , Wikesjo UM , et al. (1986). A biochemical approach to periodontal regeneration : tetracycline treatment of dentin promotes fibroblast adhesion and growth . J Periodont Res 21:330-337.

23.

Terranova VP , Hic S. , Franzetti LC , Lyall RM , Wikesjo ME (1987). A biochemical approach to periodontal regeneration-AFSCM: assay for specific cell migration. J Periodontol 58:247-257. Terranova VP , Odziemiec C. , Tweden KS , Spadone DP (1989). Repopulation of dentin surfaces by periodontal ligament cells and endothelial cells-Effects of basic fibroblast growth factor. J Periodontol 60:293-301.

24.

Tweden KS , Spadone DP , Terranova VP (1989). Neovascularization of surface demineralized dentin . J Periodontol 60:460-466. Wikesjo UM , Baker PJ , Christersson LA , Genco RJ , Lyall RM , Hie S. , et al. (1986). A biochemical approach to periodontal regeneration: tetracycline treatment conditions dentin surfaces. J Periodont Res 21:322-329.

25.

Yamada KM , Schlesinger DH , Kennedy DW , Pastan I. (1977). Characterization of a major fibroblast cell surface glycoprotein. Biochemistry 16:5552-5556.

26.

Zerbe GO (1979). Randomization analysis of the completely randomized design extended to growth and response curves. J Am Statist Assoc 74:215-221.