Effects of hydrostatic pressure on cytoskeleton and BMP-2, TGF-beta, SOX-9 production in rat temporomandibular synovial fibroblasts.
OBJECTIVE
Recent experimental evidence has suggested that pressure may play an important role in the pathogenesis of arthritic diseases such as temporomandibular disorders (TMDs), rheumatic diseases and osteoarthritis. This study examines the effects of hydrostatic pressure (HP) on cytoskeleton and protein production of bone morphogenetic protein-2 (BMP-2), transforming growth factor-beta (TGF-beta) and the SRY HMG box related gene 9 (SOX-9) in synovial fibroblasts (SFs) of rat temporomandibular joint (TMJ).


METHODS
SFs derived from rat TMJ were grown to confluence in Dulbecco's modified Eagle medium supplemented with 15% fetal calf serum. The monolayer of SFs was subjected to different HPs (0, 30, 60, and 90kPa) by an in-house designed pressure chamber for 12h. Changes of cell morphology were observed by fluorescent microscope. Production of TGF-beta, BMP-2 and SOX-9 was examined by immunocytochemical assay and western blot.


RESULTS
Compared with the untreated control, the cellular actin configuration of SFs became elongated and more intense F-actin stress fiber staining was observed after HP loading. Exposure of SFs to HP for 12h resulted in significant up-regulation of BMP-2 by 46, 54, and 66% at 30, 60, and 90kPa, respectively, whilst TGF-beta increased by 11, 19, and 28% at 30, 60, and 90kPa, respectively. HP also induced the increase of SOX-9 by 72% at 30kPa and 83% at 60kPa, but only 54% at 90kPa.


CONCLUSIONS
The obtained data suggest that HP induced the alteration of cytoskeleton and bone-morphogenetic-related proteins' production of SFs, which may influence the pathological condition of TMDs.
Wu
M-J
MJ
Department of Oral and Maxillofacial Surgery, Hospital of Stomatology, Zhejiang University, Hangzhou, China.
Gu
Z-Y
ZY
Sun
W
W
D000199:Actins / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006874:Hydrostatic Pressure; D007150:Immunohistochemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D055758:SOX9 Transcription Factor; D013583:Synovial Membrane / Q000378:metabolism / Q000473:pathology; D013704:Temporomandibular Joint / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*
D000199:Actins; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D006609:High Mobility Group Proteins; D055758:SOX9 Transcription Factor; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1