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Abstract
Objective
This study examines the effects of tumor necrosis factor (TNF) blockade on markers of bone metabolism in peripheral blood from active rheumatoid arthritis (RA) patients.
Methods
Eighteen patients (16 women, 2 men) aged 50 years (range 37-63 years), with persistently active RA (mean disease duration 7 years) were studied. Most took methotrexate (mean dose 12.5 mg) and all except one received corticosteroid (mean dose 5.7 mg). Four were treated with etanercept, eight received adalimumab and six received infliximab. Before and six months after taking TNF blockers, blood was sampled to obtain peripheral blood mononuclear cells (PBMCs), and serum bone turnover markers and acute phase reactants were measured. PBMCs were seeded and cultured to produce osteoblastic lineage cells and osteoclasts.
Results
The formation of calcified nodules by osteoblastic lineage cells from PBMC increased from 205.7±196.3 μ mol/well at the baseline to 752.5±671.9 μ mol/well after TNF blockade (p＜0.024). The serum levels of bone formation markers, including bone specific alkaline phosphatase and osteocalcin also increased. The number of circulating osteoclasts and area of bone resorption pits made by osteoclasts were reduced after TNF blockade.
Conclusion
The activity of circulating osteoblastic lineage cells increased after TNF blockade, whereas peripheral osteoclastogenesis tended to be suppressed. This is the first study of cultured human peripheral osteoblastic lineage cells in RA patients. Given that peripheral bone formation is difficult to study using radiologic methods, culture of these cells may provide a new modality for studying bone metabolism in RA.
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Figure 1.
Calcified nodule formation (a) and bone resorption pits (b) before and after tumor necrosis factor (TNF) blocker treatment. The multinucleated osteoclasts which are responsible for bone resorption is shown in left upper corner. Patients 1∼3 were treated with adalimumab, patients 4∼5 with etanercept and patients 6∼7 with infliximab. TRAP staining, 200×.
Figure 2.
Flow cytometry analysis of cells from peripheral blood to sort for both osteocalcin, bone specific alkaline phosphatase (BSALP)-positive cells. Cells positive for osteocalcin (OC) were stained with phycoerythrin (PE) and cells positive for BSALP (AP) were conjugated with peridinin chlorophyll protein (PerCP). Cells positive for both osteocalcin and BSALP were turned out to be less than 1%. UL: upper left, UR: upper right, LL: lower left, LR: lower right, Quad: quadrant.
Table 1.
Demographics of patients enrolled in the study
Table 2.
Changes in disease activities and biochemical markers of bone turnover after TNF blocker treatment
Values are presented as mean±standard deviation. DAS28 is Disease Activity Score based on 28 joints for rheumatoid arthritis patients [8]. TNF: tumor necrosis factor, ESR: erythrocyte sedimentation rate, CRP: C-reactive protein, IL: interleukin, BSALP: bone specific alkaline phosphatase, CTX: c-terminal telopeptide.
Table 3.
Changes in disease activities and biochemical markers of bone turnover in DAS28 responders after TNF blocker treatment
Values are presented as mean±standard deviation. DAS28 is Disease Activity Score based on 28 joints for rheumatoid arthritis patients [8]. TNF: tumor necrosis factor, ESR: erythrocyte sedimentation rate, CRP: C-reactive protein, IL: interleukin, BSALP: bone specific alkaline phosphatase, CTX: c-terminal telopeptide.
Table 4.
Changes in optical densities of osteoblasts, number of osteoclasts and area of bone resorption pits in addition to disease activities and biochemical markers of bone turnover in DAS28 non-responders after TNF blocker treatment
Values are presented as mean±standard deviation. DAS28 is Disease Activity Score based on 28 joints for rheumatoid arthritis patients [8]. TNF: tumor necrosis factor, ESR: erythrocyte sedimentation rate, CRP: C-reactive protein, IL: interleukin, BSALP: bone specific alkaline phosphatase, CTX: c-terminal telopeptide.
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