Effect of hydrostatic pressure of various magnitudes on osteoarthritic chondrocytes exposed to IL-1beta.
OBJECTIVE
Several in vitro studies have shown the importance of mechanical compression or hydrostatic pressure (HP) as a modulator of cartilage metabolism. The present study was undertaken to evaluate the in vitro effects of cyclical low HP (1-5 MPa) and continuous high HP (24 MPa) applied in the presence or absence of interleukin (IL)-1beta on human osteoarthritis (OA) chondrocytes.


METHODS
Chondrocytes obtained from OA cartilage were cultivated for 48 h and then exposed to pressurization in the presence or absence of IL-1beta. After pressurization, the culture medium was collected to detect the amount of proteoglycans (PG) and nitric oxide (NO) and the chondrocytes were immediately fixed for transmission electron microscopy (TEM) and processed for immunocytochemistry to localize the inducible nitric oxide synthase (iNOS).


RESULTS
A significant increase in the level of PG and a small, non-significant, decrease in NO production were observed upon exposure to cyclical low HP. On the other hand, exposure to continuous high HP resulted in a significant decrease in the PG levels and a significant increase in NO production. The presence of IL-1beta led to a significant decrease in PG levels as well as a significant increase in NO production. The cyclical low HP did not increase the PG levels significantly but caused a statistically significant decrease in NO production in cultures damaged with IL-1beta. The continuous high HP in chondrocyte cultures stimulated with IL-1beta did not significantly decrease PG production, but significantly increased NO production. The results concerning metabolic production were further confirmed by morphological findings obtained by TEM and immunocytochemical studies.


CONCLUSIONS
The findings of this study confirmed that the response of chondrocytes varies with magnitude and frequency of HP. These findings are important to understand aetiopathogenetic mechanisms of OA and to find out which type of physical activity may be best suited for the prevention and therapy of OA.
Fioravanti
Antonella
A
Department of Clinical Medicine & Immunological Sciences, Rheumatology Unit, University of Siena, Siena, Italy. foravanti7@virgilio.it
Collodel
Giulia
G
Petraglia
Angela
A
Nerucci
Fabiola
F
Moretti
Elena
E
Galeazzi
Mauro
M
D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D006874:Hydrostatic Pressure; D007150:Immunohistochemistry; D053583:Interleukin-1beta / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism; D010003:Osteoarthritis / Q000378:metabolism* / Q000503:physiopathology; D011509:Proteoglycans / Q000378:metabolism
D053583:Interleukin-1beta; D011509:Proteoglycans; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase