Human periodontal ligament cells secrete macrophage colony-stimulating factor in response to tumor necrosis factor-alpha in vitro.
BACKGROUND
Human periodontal ligament (HPDL) cells may support osteoclastogenesis by expressing receptor activator of nuclear factor-kappa B ligand (RANKL) in response to periopathogenic factors and inflammatory cytokines. Because osteoclastogenesis requires the presence of macrophage colony-stimulating factor (M-CSF), we examined whether HPDL cells secrete M-CSF in response to tumor necrosis factor-alpha (TNF-alpha).


METHODS
Cultured HPDL cells were treated with TNF-alpha in serum-free condition. The expression of M-CSF and RANKL was determined by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Inhibitors and anti-TNF receptor (TNFR) neutralizing antibodies were used for the inhibitory experiments. A migration assay was performed.


RESULTS
TNF-alpha upregulated M-CSF and RANKL in HPDL cells. The effect on M-CSF expression could be partially blocked by pyrrolidine-dithiocarbamate ammonium salt and LY294002 but not by NS398. Neutralizing antibody to TNFR1 could diminish the effect of TNF-alpha. In addition, TNF-treated culture medium exhibited chemotactic effect for RAW264.7.


CONCLUSIONS
HPDL cells are capable of secreting M-CSF and expressing RANKL in response to TNF-alpha. The upregulation of M-CSF is possibly one of the mechanisms essential for periodontal tissue destruction in response to inflammatory cytokines. The upregulation is partly through nuclear factor-kappa B (NF-kappaB) and phosphatidylinositol 3'-kinase and possibly involves TNFR1.
Yongchaitrakul
Tussanee
T
Department of Anatomy, Faculty of Dentistry, Chulalongkorn University, Pathumwan, Bangkok, Thailand.
Lertsirirangson
Kanokporn
K
Pavasant
Prasit
P
D000293:Adolescent; D000328:Adult; D000704:Analysis of Variance; D015153:Blotting, Western / Q000379:methods; D002352:Carrier Proteins / Q000378:metabolism*; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D016173:Macrophage Colony-Stimulating Factor / Q000096:biosynthesis*; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009578:Nitrobenzenes / Q000494:pharmacology; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000037:antagonists & inhibitors / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000037:antagonists & inhibitors; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013449:Sulfonamides / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D002352:Carrier Proteins; D016861:Cyclooxygenase Inhibitors; D008562:Membrane Glycoproteins; D009578:Nitrobenzenes; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D013449:Sulfonamides; C505686:TNFRSF11A protein, human; C505688:TNFSF11 protein, human; D014409:Tumor Necrosis Factor-alpha; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D016173:Macrophage Colony-Stimulating Factor