Serum from children with polyarticular juvenile idiopathic arthritis (pJIA) inhibits differentiation, mineralization and may increase apoptosis of human osteoblasts "in vitro".
We examined the effects of polyarticular juvenile idiopathic arthritis (pJIA) serum on proliferation, differentiation, mineralization, and apoptosis of human osteoblast cells (hOb) in culture. The hOb were cultured with 10% serum from active pJIA and healthy controls (CT) and were tested for DNA synthesis, alkaline phosphatase (AP) activity, osteocalcin (OC) secretion, calcium levels, caspase 3 activity, and DNA fragmentation. None of the patients had used glucocorticoids for at least 1 month before the study, or any other drug that can affect bone mineral metabolism. Human inflammatory cytokine levels (IL-6, IL-8, IL-10, IL-1beta, TNF-alpha, and IL-12p70) were measured in pJIA and CT sera. Low levels of AP activity was observed in pJIA cultures compared with CT cultures (67.16 +/- 53.35 vs 100.11 +/- 50.64 mumol p-nitrophenol/h(-1) mg(-1) protein, P = 0.008). There was also a significant decrease in OC secretion (9.23 +/- 5.63 vs 12.82 +/- 7.02 ng/mg protein, P = 0.012) and calcium levels (0.475 +/- 0.197 vs 0.717 +/- 0.366 mmol/l, P = 0.05) in pJIA hOb cultures. No difference was observed in cell proliferation (323.56 +/- 108.23 vs 328.91 +/- 88.03 dpm/mg protein, P = 0.788). Osteoblasts cultured with JIA sera showed lower levels of DNA and increased fragmentation than osteoblasts cultured with CT sera. pJIA sera showed higher IL-6 values than CT (21.44 +/- 9.31 vs 3.58 +/- 2.38 pg/ml, P < 0.001), but no difference was observed related to IL-8, IL-10, IL-1beta, TNF-alpha, and IL-12p70 between pJIA and controls. This study suggests that serum from children with pJIA inhibits differentiation, mineralization and may increase apoptosis of hOb cultures, and inflammatory cytokines such as IL-6 might be a mechanism in this find. These results may represent an alternative therapeutic target for prevention and treatment of bone loss in JIA.
Caparbo
Valéria F
VF
Rheumatology Division, School of Medicine, University of São Paulo, São Paulo, Brazil.
Prada
Flávia
F
Silva
Clóvis A A
CA
Regio
Paula L
PL
Pereira
Rosa M R
RM
D000469:Alkaline Phosphatase / Q000378:metabolism; D017209:Apoptosis; D001171:Arthritis, Juvenile / Q000097:blood* / Q000503:physiopathology; D002113:Calcification, Physiologic; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D002648:Child; D016207:Cytokines / Q000378:metabolism; D004247:DNA / Q000032:analysis; D053938:DNA Fragmentation; D005260:Female; D006801:Humans; D008297:Male; D010006:Osteoblasts / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D015675:Osteocalcin / Q000378:metabolism; D044967:Serum / Q000378:metabolism*
D016207:Cytokines; D015675:Osteocalcin; D004247:DNA; D000469:Alkaline Phosphatase; D002118:Calcium