Impaired degradation of matrix collagen in human gingival fibroblasts by the antiepileptic drug phenytoin

Takahiro Kato, Nobuo Okahashi, Shinji Kawai, Takafumi Kato, Hiroaki Inaba, Ichijiro Morisaki, Atsuo Amano

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Background: Gingival overgrowth (GO) is a serious adverse effect associated with the administration of phenytoin (PHT), with PHT-induced GO characterized by a massive accumulation of extracellular matrix components, especially collagen, in gingival connective tissues. However, the etiology of such collagen accumulation is still largely unknown. We examined the effects of PHT on the collagen degradation process leading to collagen accumulation in human gingival fibroblasts (HGF). Methods: HGFs were cultured with various concentrations of PHT and viable cell numbers and collagen amounts were determined. Gene and protein expressions of matrix metalloproteinases (MMP) and tissue inhibitors of MMPs (TIMP) were quantified with reverse transcription-polymerase chain reaction (RT-PCR) analyses and Western blotting, respectively. Cellular endocytosis of collagen was assayed using flow-cytometric analysis. The effects of PHT on extracellular signal-regulated kinase 1/2 (ERK1/2) and inhibitor κB-α (IκB-α) were assayed. Results: The proliferation of HGFs was not affected by PHT, whereas it significantly increased collagen accumulation. Further, the expressions of MMP-1, -2, and -3 were markedly suppressed by PHT, whereas that of TIMP-1 was induced in a dose- and time-dependent manner. PHT also markedly prevented collagen endocytosis by HGFs, which was associated with the suppression of α2β1-integrin expression. In addition, the phosphorylation of ERK1/2 and IκB-α degradation were suppressed by PHT. Conclusions: These results suggest that PHT causes an impaired degradation of collagen by suppression of enzymatic degradation with MMPs/TIMP-1 and α2β1-integrin-mediated endocytosis. Those alterations are likely mediated through the cellular signaling pathways of ERK1/2 and nuclear factor κB. These synergistic effects may cause collagen accumulation, leading to GO.

Original languageEnglish
Pages (from-to)941-950
Number of pages10
JournalJournal of Periodontology
Volume76
Issue number6
DOIs
Publication statusPublished - Jun 2005
Externally publishedYes

Fingerprint

Phenytoin
Anticonvulsants
Collagen
Fibroblasts
Gingival Overgrowth
Matrix Metalloproteinase Inhibitors
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinase 1
Endocytosis
Matrix Metalloproteinases
Integrins
Matrix Metalloproteinase 1
Matrix Metalloproteinase 2
Connective Tissue
Reverse Transcription
Extracellular Matrix
Cell Count
Western Blotting
Phosphorylation
Gene Expression

Keywords

  • Collagen diseases
  • Fibroblasts, gingival
  • Gingival hyperplasia/etiology
  • Metalloproteinases, matrix
  • Phenytoin/adverse effects

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Impaired degradation of matrix collagen in human gingival fibroblasts by the antiepileptic drug phenytoin. / Kato, Takahiro; Okahashi, Nobuo; Kawai, Shinji; Kato, Takafumi; Inaba, Hiroaki; Morisaki, Ichijiro; Amano, Atsuo.

In: Journal of Periodontology, Vol. 76, No. 6, 06.2005, p. 941-950.

Research output: Contribution to journalArticle

Kato, Takahiro ; Okahashi, Nobuo ; Kawai, Shinji ; Kato, Takafumi ; Inaba, Hiroaki ; Morisaki, Ichijiro ; Amano, Atsuo. / Impaired degradation of matrix collagen in human gingival fibroblasts by the antiepileptic drug phenytoin. In: Journal of Periodontology. 2005 ; Vol. 76, No. 6. pp. 941-950.
@article{c886634109384078a44903699da1dd43,
title = "Impaired degradation of matrix collagen in human gingival fibroblasts by the antiepileptic drug phenytoin",
abstract = "Background: Gingival overgrowth (GO) is a serious adverse effect associated with the administration of phenytoin (PHT), with PHT-induced GO characterized by a massive accumulation of extracellular matrix components, especially collagen, in gingival connective tissues. However, the etiology of such collagen accumulation is still largely unknown. We examined the effects of PHT on the collagen degradation process leading to collagen accumulation in human gingival fibroblasts (HGF). Methods: HGFs were cultured with various concentrations of PHT and viable cell numbers and collagen amounts were determined. Gene and protein expressions of matrix metalloproteinases (MMP) and tissue inhibitors of MMPs (TIMP) were quantified with reverse transcription-polymerase chain reaction (RT-PCR) analyses and Western blotting, respectively. Cellular endocytosis of collagen was assayed using flow-cytometric analysis. The effects of PHT on extracellular signal-regulated kinase 1/2 (ERK1/2) and inhibitor κB-α (IκB-α) were assayed. Results: The proliferation of HGFs was not affected by PHT, whereas it significantly increased collagen accumulation. Further, the expressions of MMP-1, -2, and -3 were markedly suppressed by PHT, whereas that of TIMP-1 was induced in a dose- and time-dependent manner. PHT also markedly prevented collagen endocytosis by HGFs, which was associated with the suppression of α2β1-integrin expression. In addition, the phosphorylation of ERK1/2 and IκB-α degradation were suppressed by PHT. Conclusions: These results suggest that PHT causes an impaired degradation of collagen by suppression of enzymatic degradation with MMPs/TIMP-1 and α2β1-integrin-mediated endocytosis. Those alterations are likely mediated through the cellular signaling pathways of ERK1/2 and nuclear factor κB. These synergistic effects may cause collagen accumulation, leading to GO.",
keywords = "Collagen diseases, Fibroblasts, gingival, Gingival hyperplasia/etiology, Metalloproteinases, matrix, Phenytoin/adverse effects",
author = "Takahiro Kato and Nobuo Okahashi and Shinji Kawai and Takafumi Kato and Hiroaki Inaba and Ichijiro Morisaki and Atsuo Amano",
year = "2005",
month = "6",
doi = "10.1902/jop.2005.76.6.941",
language = "English",
volume = "76",
pages = "941--950",
journal = "Journal of Periodontology",
issn = "0022-3492",
publisher = "American Academy of Periodontology",
number = "6",

}

TY - JOUR

T1 - Impaired degradation of matrix collagen in human gingival fibroblasts by the antiepileptic drug phenytoin

AU - Kato, Takahiro

AU - Okahashi, Nobuo

AU - Kawai, Shinji

AU - Kato, Takafumi

AU - Inaba, Hiroaki

AU - Morisaki, Ichijiro

AU - Amano, Atsuo

PY - 2005/6

Y1 - 2005/6

N2 - Background: Gingival overgrowth (GO) is a serious adverse effect associated with the administration of phenytoin (PHT), with PHT-induced GO characterized by a massive accumulation of extracellular matrix components, especially collagen, in gingival connective tissues. However, the etiology of such collagen accumulation is still largely unknown. We examined the effects of PHT on the collagen degradation process leading to collagen accumulation in human gingival fibroblasts (HGF). Methods: HGFs were cultured with various concentrations of PHT and viable cell numbers and collagen amounts were determined. Gene and protein expressions of matrix metalloproteinases (MMP) and tissue inhibitors of MMPs (TIMP) were quantified with reverse transcription-polymerase chain reaction (RT-PCR) analyses and Western blotting, respectively. Cellular endocytosis of collagen was assayed using flow-cytometric analysis. The effects of PHT on extracellular signal-regulated kinase 1/2 (ERK1/2) and inhibitor κB-α (IκB-α) were assayed. Results: The proliferation of HGFs was not affected by PHT, whereas it significantly increased collagen accumulation. Further, the expressions of MMP-1, -2, and -3 were markedly suppressed by PHT, whereas that of TIMP-1 was induced in a dose- and time-dependent manner. PHT also markedly prevented collagen endocytosis by HGFs, which was associated with the suppression of α2β1-integrin expression. In addition, the phosphorylation of ERK1/2 and IκB-α degradation were suppressed by PHT. Conclusions: These results suggest that PHT causes an impaired degradation of collagen by suppression of enzymatic degradation with MMPs/TIMP-1 and α2β1-integrin-mediated endocytosis. Those alterations are likely mediated through the cellular signaling pathways of ERK1/2 and nuclear factor κB. These synergistic effects may cause collagen accumulation, leading to GO.

AB - Background: Gingival overgrowth (GO) is a serious adverse effect associated with the administration of phenytoin (PHT), with PHT-induced GO characterized by a massive accumulation of extracellular matrix components, especially collagen, in gingival connective tissues. However, the etiology of such collagen accumulation is still largely unknown. We examined the effects of PHT on the collagen degradation process leading to collagen accumulation in human gingival fibroblasts (HGF). Methods: HGFs were cultured with various concentrations of PHT and viable cell numbers and collagen amounts were determined. Gene and protein expressions of matrix metalloproteinases (MMP) and tissue inhibitors of MMPs (TIMP) were quantified with reverse transcription-polymerase chain reaction (RT-PCR) analyses and Western blotting, respectively. Cellular endocytosis of collagen was assayed using flow-cytometric analysis. The effects of PHT on extracellular signal-regulated kinase 1/2 (ERK1/2) and inhibitor κB-α (IκB-α) were assayed. Results: The proliferation of HGFs was not affected by PHT, whereas it significantly increased collagen accumulation. Further, the expressions of MMP-1, -2, and -3 were markedly suppressed by PHT, whereas that of TIMP-1 was induced in a dose- and time-dependent manner. PHT also markedly prevented collagen endocytosis by HGFs, which was associated with the suppression of α2β1-integrin expression. In addition, the phosphorylation of ERK1/2 and IκB-α degradation were suppressed by PHT. Conclusions: These results suggest that PHT causes an impaired degradation of collagen by suppression of enzymatic degradation with MMPs/TIMP-1 and α2β1-integrin-mediated endocytosis. Those alterations are likely mediated through the cellular signaling pathways of ERK1/2 and nuclear factor κB. These synergistic effects may cause collagen accumulation, leading to GO.

KW - Collagen diseases

KW - Fibroblasts, gingival

KW - Gingival hyperplasia/etiology

KW - Metalloproteinases, matrix

KW - Phenytoin/adverse effects

UR - http://www.scopus.com/inward/record.url?scp=21344453286&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=21344453286&partnerID=8YFLogxK

U2 - 10.1902/jop.2005.76.6.941

DO - 10.1902/jop.2005.76.6.941

M3 - Article

C2 - 15948689

AN - SCOPUS:21344453286

VL - 76

SP - 941

EP - 950

JO - Journal of Periodontology

JF - Journal of Periodontology

SN - 0022-3492

IS - 6

ER -