Noriki Miyamoto

Gene expression during osteoclast-like cell formation induced by antifusion regulatory protein-1/CD98/4F2 monoclonal antibodies (MAbs): c-src is selectively induced by anti-FRP-1 MAb

Human blood monocytes can differentiate into osteoclast-like cells when they are cultured in the presence of anti-FRP-1. Messenger (mRNA) expression of markers related to osteoclasts was analyzed during differentiation of osteoclasts from monocytes. As markers related to osteoclasts, we selected cathepsin-K, carbonic anhydrase (CA) II, vacuolar H(+)-ATPase (v-ATPase), vitronectin receptor (VNR), tartrate-resistant acid phosphatase (TRAP), osteopontin (OPN), galectin-3, c-src, c-fos, and c-fms. The mRNAs other than c-src mRNA were expressed in freshly isolated monocytes or monocytes incubated with control antibody or anti-FRP-1 monoclonal antibody (MAb) for 14 days. Of these mRNAs, cathepsin-K, CA II, v-ATPase, VNR, TRAP, OPN, and c-fms mRNAs were expressed at higher levels in the osteoclast-like cells than those in monocytes cultured with control antibody. On the other hand, galectin-3 mRNA was expressed at lower levels in the osteoclast-like cells, and there was no significant difference in c-fos mRNA expression between the monocytes cultured with control antibody and anti-FRP-1 MAb. c-src mRNA could not be detected in monocytes freshly isolated or incubated with control antibody. Surprisingly, expression of c-src mRNA was induced in monocytes by anti-FRP-1 MAb and was detectable as early as 3 h after anti-FRP-1 MAb treatment, indicating that c-src is selectively induced by anti-FRP-1 MAb treatment. Furthermore, the osteoclast-like cells expressed calcitonin receptor. Receptor activator of NF-kappaB (RANK) mRNA was detectable in freshly isolated monocytes or monocytes cultured with control antibody or anti-FRP-1 MAbs. Maximal expression of RANK was observed in osteoclast-like cells. On the other hand, no receptor activator of NF-KB ligand (RANKL) mRNA was detectable in any of the samples, suggesting that anti-FRP-1 mAb can induce osteoclast-like cells from blood monocytes without RANKL.

Prevalence and risk factors for knee osteoarthritis in elderly Japanese men and women

BackgroundThe aims of the present study were to examine the prevalence and risk factors for knee osteoarthritis in elderly Japanese men and women.MethodsWe examined 598 of the 1513 inhabitants of Miyagawa village aged ≥ 65 years (393 women, 205 men). Baseline data, obtained with standard questionnaires, included information on age, past history, sports activity, working, knee pain, smoking, and intakes of alcohol and milk. Bone mineral density of the forearm was measured using dual energy X-ray absorptiometry. Anteroposterior radiographs of both knees were graded for osteoarthritis using the Kellgren-Lawrence grading system. Definite osteoarthritis was defined as a grade of 2 or higher. We used logistic regression analysis by the stepwise method to determine the risk factors for radiographic knee osteoarthritis.ResultsThe prevalence of definite radiographic knee osteoarthritis was 30.0% overall: 17.7% in men and 36.5% in women. The prevalence of symptomatic knee osteoarthritis was 21.2% overall: 10.7% in men and 26.7% in women. There were significant differences in the risk of radiographic knee osteoarthritis with body mass index (BMI), sex, age, and bone mineral density (BMD).ConclusionsThe prevalence of definite radiographic knee osteoarthritis was 30.0% and that of symptomatic knee osteoarthritis was 21.2%. We found that higher BMI, female sex, older age, and higher BMD were significantly associated with an increased risk for radiographic knee osteoarthritis.

Human osteosarcoma-derived cell lines produce soluble factor(s) that induces differentiation of blood monocytes to osteoclast-like cells

When monocytes were cocultured with human osteosarcoma-derived cells (HOS cells), multinucleated giant cell formation of monocytes was induced. Intriguingly, even when a filter was interposed between monocytes and HOS cells, polykaryocytes also appeared. The multinucleated giant cells have characters similar to osteoclast-like cells. These findings indicate that soluble factor(s) secreted from HOS cells play an important role in polykaryocyte formation from monocytes. Twelve cloned cells were established from HSOS-1 cells and their capacities of inducing osteoclasts were investigated. Three cloned cells inducing nos. 4 and 9 had an ability of inducing osteoclasts (multinucleated giant cells, TRAP, calcitonin receptor and c-src mRNAs, osteoresorbing activity), and three cells, including nos. 1 and 5, did not show the ability. HOS cells and the cloned cells expressed several cytokine mRNAs. M-CSF was detected in the culture fluids of HOS cells, which also expressed RANK and RANK/ODF/OPGL mRNAs. Intriguingly, HOS cells secreting a soluble osteoclast inducing factors(s) expressed TNF-alpha converting enzyme mRNA. Furthermore, OCIF/OPG inhibited HOS cell-induced osteoclastogenesis and soluble RANKL could be detected in the culture fluids of HOS cells expressing TACE, suggesting that one of soluble osteoclast-inducing factor(s) is soluble RANKL. When blood monocytes were indirectly cocultured with HSOS-1 cells or cloned no. 9 cells in the presence of OCIF for 14 days, HOS cell-mediated osteoclastogenesis was suppressed, indicating that RANK-RANKL system is involved in the HOS cell-mediated osteoclastogenesis.

Longitudinal Magnetic Resonance Imaging of Pseudotumors Following Metal-on-Metal Total Hip Arthroplasty

The purpose of the study was to determine the natural history of pseudotumors following metal-on-metal total hip arthroplasty (THA) using magnetic resonance imaging (MRI). Initial MRI was conducted at a mean of 36months postoperatively. Follow-up MRI was performed at a mean of 20months after the detection of 24 asymptomatic pseudotumors. Pseudotumor size was determined on MRI. The mean pseudotumor size changed from 729mm(2) to 877mm(2). Pseudotumors increased in size in eight and decreased in six. Ten hips showed no changes. The bigger the pseudotumor size, the more likely the size would increase. In conclusion, pseudotumors frequently change in size. A single MRI study in the clinical decision-making process should be avoided and a longitudinal study should be performed.

The relationships among hemostatic markers, the withdrawal of fondaparinux due to a reduction in hemoglobin and deep vein thrombosis in Japanese patients undergoing major orthopedic surgery.

BACKGROUND The relationships among the hemostatic markers, the development of deep vein thrombosis (DVT) and the withdrawal of fondaparinux due to a reduction in the hemoglobin levels were examined. METHODS Two-hundred twenty-one Japanese patients who underwent major orthopedic surgery and were treated with 1.5mg of fondaparinux instead of 2.5mg of fondaparinux were studied. Forty-seven of 221 patients discontinued fondaparinux treatment (withdrawal group) and 37 patients developed DVT. RESULTS The age, frequency of total knee arthroplasty (TKA), withdrawal of fondaparinux, reduction of hemoglobin and the plasma levels of soluble fibrin (SF), D-dimer and fibrinogen and fibrin degradation product (FDP) on day 1 after the operation were significantly higher in the patients with DVT. Elevated SF, D-dimer or FDP levels were associated with the risk for DVT. The age, frequency of TKA or DVT, anti-Xa activity and the creatinine, FDP and D-dimer levels were significantly higher in the withdrawal group. An anti-Xa level >0.33 mg/l and an elevated D-dimer or FDP level were associated with the risk of withdrawal. CONCLUSION The age and SF levels, TKA and withdrawal of fondaparinux were related to the risk of DVT, and the anti-Xa activity, creatinine level and DVT were related to the risk of withdrawal of fondaparinux due to a reduction in hemoglobin.

Comparison of bone mineral density among residents of a mountain village and a fishing village in Japan

Objective. To compare the bone mineral density of residents of a mountain village with that of residents of a fishing village in Mie Prefecture, Japan. Methods. Microdensitometry was used to measure bone mineral density of the second metacarpal bone of 202 participants living in a mountain village and of 852 participants living in a fishing village to identify contributory factors for osteoporosis. The participants were interviewed using a questionnaire on alcohol consumption, fish intake, milk intake, and daily activity. Results. Analysis of covariance revealed that bone mineral density was significantly higher among the participants living in a fishing village than among those living in a mountain village (2.5–2.9 versus 2.1–2.7 mmAl; p<0.001). A higher proportion of women in the fishing village than of those in the mountain village consumed alcohol (17% versus 10%; p<0.05). Conclusion. Nutrition may be a contributory factor to the lower incidence of osteoporosis among residents of the fishing village compared with those of the mountain village.

Genome-wide Association Study of Idiopathic Osteonecrosis of the Femoral Head

Idiopathic osteonecrosis of the femoral head (IONFH) is an ischemic disorder that causes bone necrosis of the femoral head, resulting in hip joint dysfunction. IONFH is a polygenic disease and steroid and alcohol have already known to increase its risk; however, the mechanism of IONFH remains to be elucidated. We performed a genome-wide association study using ~60,000 subjects and found two novel loci on chromosome 20q12 and 12q24. Big data analyses identified LINC01370 as a candidate susceptibility gene in the 20q12 locus. Stratified analysis by IONFH risk factors suggested that the 12q24 locus was associated with IONFH through drinking capacity. Our findings would shed new light on pathophysiology of IONFH.