Effect of enamel matrix derivative on periodontal ligament cells in vitro is diminished by Porphyromonas gingivalis.
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 (delta fimA) or gingipains (delta rgpA delta rgpB, delta kgp, and delta rgpA delta rgpB delta kgp) 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-beta1 (TGF-beta1) 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 delta fimA, delta rgpA delta rgpB, and delta kgp showed the same effects. Further, each of these organisms diminished the expression of TGF-beta1 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 delta rgpA delta rgpB delta kgp, which showed a negligible effect in all of the assays with EMD-stimulated PDL cells.


CONCLUSIONS
These results suggest that P. gingivalis diminishes the effect of EMD on PDL cells in vitro through a cooperative action of gingipains.
Department of Oral Frontier Biology, Osaka University Graduate School of Dentistry, Suita-Osaka, Japan.
Hiroaki
H
Inaba
Shinji
S
Kawai
Koji
K
Nakayama
Nobuo
N
Okahashi
Atsuo
A
Amano
D018829:Adhesins, Bacterial; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000494:pharmacology; D003598:Cytoskeletal Proteins / Q000009:adverse effects; D003746:Dental Enamel Proteins / Q000037:antagonists & inhibitors* / Q000494:pharmacology*; D000080867:Gingipain Cysteine Endopeptidases; D006133:Growth Substances / Q000096:biosynthesis; D006388:Hemagglutinins / Q000494:pharmacology; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D051419:Paxillin; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism / Q000382:microbiology*; D010750:Phosphoproteins / Q000009:adverse effects; D016966:Porphyromonas gingivalis / Q000201:enzymology / Q000472:pathogenicity*; D014945:Wound Healing / Q000187:drug effects
D018829:Adhesins, Bacterial; D003598:Cytoskeletal Proteins; D003746:Dental Enamel Proteins; D000080867:Gingipain Cysteine Endopeptidases; D006133:Growth Substances; D006388:Hemagglutinins; C496061:PXN protein, human; D051419:Paxillin; D010750:Phosphoproteins; C109069:enamel matrix proteins; D020928:Mitogen-Activated Protein Kinases; D003546:Cysteine Endopeptidases