Effect of enamel matrix derivative on periodontal ligament cells in vitro is deminished by porphyromonas gingivalis

Hiroaki Inaba, Shinji Kawai, Koji Nakayama, Nobuo Okahashi, Atsuo Amano

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Background: Enamel matrix derivative (EMD) has been shown to possess a mitogenic effect to induce effective periodontal regeneration, however, it is unclear whether periodontal pathogens can modulate the effect of EMD. The present study examined the influence of Porphyromonas gingivalis on EMD-stimulated periodontal ligament (PDL) cells. Methods: P. gingivalis ATCC33277 and its mutants deficient in fimbriae (ΔfimA) or gingipains (ΔrgpAΔrgpB, Δkgp, and ΔrgpAΔrgpBΔkgp) were employed. PDL cells were grown on EMD-coated dishes and infected with P. gingivalis wild strain or a mutant. Cell migration and proliferation were then evaluated with an in vitro wound healing assay. The expression of transforming growth factor-β1 (TGF-β1) and insulin-like growth factor I (IGF-I) mRNA by PDL cells was examined. Further, the degradation and phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2 as well as paxillin in infected PDL cells were estimated using Western blot analysis. Results: P. gingivalis ATCC33277 inhibited the migration and proliferation of PDL cells on EMD-coated dishes, and the mutants ΔfimA, ΔrgpAΔrgpB, and Δkgp showed the same effects. Further, each of these organisms diminished the expression of TGF-β1 and IGF-I mRNA, as well as the phosphorylation of ERK1/2 from EMD-stimulated PDL cells. In addition, total paxillin protein was markedly degraded by both the wild-type strain and each of the mutants except for ΔrgpAΔrgpBΔkgp, which showed a negligible effect in all of the assays with EMD-stimulated PDL cells. Conclusion: These results suggest that P. gingivalis diminishes the effect of EMD on PDL cells in vitro through a cooperative action of gingipains.

Original languageEnglish
Pages (from-to)858-865
Number of pages8
JournalJournal of Periodontology
Volume75
Issue number6
DOIs
Publication statusPublished - Jun 2004
Externally publishedYes

Fingerprint

Periodontal Ligament
Porphyromonas gingivalis
Dental Enamel
Paxillin
Transforming Growth Factors
Insulin-Like Growth Factor I
Phosphorylation
Messenger RNA
Mitogen-Activated Protein Kinase 3
In Vitro Techniques
Mitogen-Activated Protein Kinase 1
Wound Healing
Cell Movement
Regeneration
Western Blotting
Cell Proliferation

Keywords

  • Cells, periodontal
  • Enamel matrix derivative
  • Periodontal ligament/anatomy and histology
  • Periodontal regeneration
  • Porphyromonas gingivalis

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Effect of enamel matrix derivative on periodontal ligament cells in vitro is deminished by porphyromonas gingivalis. / Inaba, Hiroaki; Kawai, Shinji; Nakayama, Koji; Okahashi, Nobuo; Amano, Atsuo.

In: Journal of Periodontology, Vol. 75, No. 6, 06.2004, p. 858-865.

Research output: Contribution to journalArticle

Inaba, Hiroaki ; Kawai, Shinji ; Nakayama, Koji ; Okahashi, Nobuo ; Amano, Atsuo. / Effect of enamel matrix derivative on periodontal ligament cells in vitro is deminished by porphyromonas gingivalis. In: Journal of Periodontology. 2004 ; Vol. 75, No. 6. pp. 858-865.
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abstract = "Background: Enamel matrix derivative (EMD) has been shown to possess a mitogenic effect to induce effective periodontal regeneration, however, it is unclear whether periodontal pathogens can modulate the effect of EMD. The present study examined the influence of Porphyromonas gingivalis on EMD-stimulated periodontal ligament (PDL) cells. Methods: P. gingivalis ATCC33277 and its mutants deficient in fimbriae (ΔfimA) or gingipains (ΔrgpAΔrgpB, Δkgp, and ΔrgpAΔrgpBΔkgp) were employed. PDL cells were grown on EMD-coated dishes and infected with P. gingivalis wild strain or a mutant. Cell migration and proliferation were then evaluated with an in vitro wound healing assay. The expression of transforming growth factor-β1 (TGF-β1) and insulin-like growth factor I (IGF-I) mRNA by PDL cells was examined. Further, the degradation and phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2 as well as paxillin in infected PDL cells were estimated using Western blot analysis. Results: P. gingivalis ATCC33277 inhibited the migration and proliferation of PDL cells on EMD-coated dishes, and the mutants ΔfimA, ΔrgpAΔrgpB, and Δkgp showed the same effects. Further, each of these organisms diminished the expression of TGF-β1 and IGF-I mRNA, as well as the phosphorylation of ERK1/2 from EMD-stimulated PDL cells. In addition, total paxillin protein was markedly degraded by both the wild-type strain and each of the mutants except for ΔrgpAΔrgpBΔkgp, which showed a negligible effect in all of the assays with EMD-stimulated PDL cells. Conclusion: These results suggest that P. gingivalis diminishes the effect of EMD on PDL cells in vitro through a cooperative action of gingipains.",
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T1 - Effect of enamel matrix derivative on periodontal ligament cells in vitro is deminished by porphyromonas gingivalis

AU - Inaba, Hiroaki

AU - Kawai, Shinji

AU - Nakayama, Koji

AU - Okahashi, Nobuo

AU - Amano, Atsuo

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N2 - Background: Enamel matrix derivative (EMD) has been shown to possess a mitogenic effect to induce effective periodontal regeneration, however, it is unclear whether periodontal pathogens can modulate the effect of EMD. The present study examined the influence of Porphyromonas gingivalis on EMD-stimulated periodontal ligament (PDL) cells. Methods: P. gingivalis ATCC33277 and its mutants deficient in fimbriae (ΔfimA) or gingipains (ΔrgpAΔrgpB, Δkgp, and ΔrgpAΔrgpBΔkgp) were employed. PDL cells were grown on EMD-coated dishes and infected with P. gingivalis wild strain or a mutant. Cell migration and proliferation were then evaluated with an in vitro wound healing assay. The expression of transforming growth factor-β1 (TGF-β1) and insulin-like growth factor I (IGF-I) mRNA by PDL cells was examined. Further, the degradation and phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2 as well as paxillin in infected PDL cells were estimated using Western blot analysis. Results: P. gingivalis ATCC33277 inhibited the migration and proliferation of PDL cells on EMD-coated dishes, and the mutants ΔfimA, ΔrgpAΔrgpB, and Δkgp showed the same effects. Further, each of these organisms diminished the expression of TGF-β1 and IGF-I mRNA, as well as the phosphorylation of ERK1/2 from EMD-stimulated PDL cells. In addition, total paxillin protein was markedly degraded by both the wild-type strain and each of the mutants except for ΔrgpAΔrgpBΔkgp, which showed a negligible effect in all of the assays with EMD-stimulated PDL cells. Conclusion: These results suggest that P. gingivalis diminishes the effect of EMD on PDL cells in vitro through a cooperative action of gingipains.

AB - Background: Enamel matrix derivative (EMD) has been shown to possess a mitogenic effect to induce effective periodontal regeneration, however, it is unclear whether periodontal pathogens can modulate the effect of EMD. The present study examined the influence of Porphyromonas gingivalis on EMD-stimulated periodontal ligament (PDL) cells. Methods: P. gingivalis ATCC33277 and its mutants deficient in fimbriae (ΔfimA) or gingipains (ΔrgpAΔrgpB, Δkgp, and ΔrgpAΔrgpBΔkgp) were employed. PDL cells were grown on EMD-coated dishes and infected with P. gingivalis wild strain or a mutant. Cell migration and proliferation were then evaluated with an in vitro wound healing assay. The expression of transforming growth factor-β1 (TGF-β1) and insulin-like growth factor I (IGF-I) mRNA by PDL cells was examined. Further, the degradation and phosphorylation of extracellular signal-regulated kinase 1 and 2 (ERK1/2 as well as paxillin in infected PDL cells were estimated using Western blot analysis. Results: P. gingivalis ATCC33277 inhibited the migration and proliferation of PDL cells on EMD-coated dishes, and the mutants ΔfimA, ΔrgpAΔrgpB, and Δkgp showed the same effects. Further, each of these organisms diminished the expression of TGF-β1 and IGF-I mRNA, as well as the phosphorylation of ERK1/2 from EMD-stimulated PDL cells. In addition, total paxillin protein was markedly degraded by both the wild-type strain and each of the mutants except for ΔrgpAΔrgpBΔkgp, which showed a negligible effect in all of the assays with EMD-stimulated PDL cells. Conclusion: These results suggest that P. gingivalis diminishes the effect of EMD on PDL cells in vitro through a cooperative action of gingipains.

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