Akira Hozumi

Primary human bone marrow adipocytes support TNF-α-induced osteoclast differentiation and function through RANKL expression

PURPOSE In previous reports, it was demonstrated that bone marrow adipocytes were related to steroid osteoporosis through osteoclastogenesis induced by Receptor Activator of Nuclear factor κ-B Ligand (RANKL) expression. The purpose of this study was to evaluate the effect of Tumor necrosis factor-alpha (TNF-α) on RANKL expression in bone marrow adipocytes, and osteoclast differentiation supported by human bone marrow adipocytes. METHODS RANKL, osteoprotegerin (OPG), and macrophage-colony stimulating factor (M-CSF) mRNA expression in bone marrow adipocytes and their regulation by TNF-α treatment were measured by real-time RT-PCR. Co-cultures of bone marrow adipocytes and osteoclast precursors were performed with or without TNF-α, and osteoclast differentiation was evaluated morphologically and functionally. RESULTS RANKL expression and an increase in the RANKL/OPG ratio in bone marrow adipocytes were stimulated by TNF-α treatment. In co-culture of bone marrow adipocytes and osteoclast precursors with TNF-α, the number of TRAP-positive multinuclear cells and resorption cavity formations of calcium phosphate film were increased. Osteoclast differentiation was suppressed by anti-RANKL antibody treatment. In co-culture with non-cell-contact conditions, no TRAP-positive cells or resorption cavity formations were observed. CONCLUSIONS TNF-α increased RANKL expression in primary human bone marrow adipocytes. TNF-α induced the ability of bone marrow adipocytes to promote osteoclast differentiation and activity in a manner directly related to RANKL expression.

Bone marrow adipocytes support dexamethasone-induced osteoclast differentiation

The purpose of this study was to examine the ability of bone marrow adipocytes to support osteoclast differentiation in vitro. The primary bone marrow adipocytes were obtained from bone marrow fluid during prosthesis insertion. NFkappa-B ligand (RANKL), Osteoprotegerin (OPG), and macrophage colony stimulating factor (M-CSF) expressions in bone marrow adipocytes with or without dexamethasone were examined. In a co-culture system with bone marrow adipocytes and osteoclast precursor cells, osteoclast differentiation was assessed by the expression of titrate-resistant acid phosphatase (TRAP) staining. RANKL, OPG, and M-CSF mRNA expressions were confirmed in all individuals. Dexamethasone significantly induced RANKL and OPG expression. The RANKL/OPG ratio was increased by dexamethasone and was significant at 10(-7) M dexamethasone. With 10(-7) M dexamethasone, osteoclast precursor cells differentiated into multinucleated TRAP-positive cells when co-cultured with bone marrow adipocytes. The present study demonstrates for the first time that bone marrow adipocytes can support osteoclast differentiation in vitro.

Simvastatin suppresses dexamethasone-induced secretion of plasminogen activator inhibitor-1 in human bone marrow adipocytes

BackgroundOsteonecrosis of the femoral head is a common complication of high-dose glucocorticoid treatment. Intravascular thrombosis is thought to be associated with the ischemic state of the femoral head. Plasminogen activator inhibitor-1 (PAI-1) is an adipokine, which are physiologically active substances secreted from visceral and subcutaneous adipocytes. PAI-1 suppresses fibrinolysis by binding tissue-type plasminogen activator. Several reports have described the relationship between PAI-1 and steroid-induced osteonecrosis of the femoral head, and the preventive effects of lipid-lowering agents (statins) against steroid-induced osteonecrosis of the femoral head. We previously reported that adipokines and dexamethasone induced PAI-1 secretion from bone marrow adipocytes. The purpose of the present study is to examine the effects of simvastatin on PAI-1 secretion from human bone marrow adipocytes in vitro.MethodsPrimary bone marrow adipocytes were extracted from collagenase-treated bone marrow fluid obtained from the femoral necks of 40 patients (6 men, 34 women; age range, 52-81 years) undergoing hip joint replacement surgery. After suspended culture with or without dexamethasone or simvastatin, PAI-1 mRNA expression was assessed by real-time RT-PCR. Total PAI-1 protein secretion in culture medium was assessed by enzyme-linked immunosorbent assay.ResultsPAI-1 mRNA expression was up-regulated by 388% (P = 0.002) with dexamethasone, and down-regulated by 45% (P = 0.002) with simvastatin, as compared to control levels. Dexamethasone increased total PAI-1 secretion by 166% (P = 0.001) and simvastatin decreased total PAI-1 secretion by 64% (P = 0.002). No significant changes were observed in adiponectin mRNA expression and secretion by dexamethasone and simvastatin, while pre-treatment with simvastatin reversed dexamethasone induced PAI-1 secretion by 89%, as compared to control levels.ConclusionThe present study confirmed the suppressive effects of simvastatin on PAI-1 expression and secretion from bone marrow adipocytes. Furthermore, pre-treatment with simvastatin reversed dexamethasone induced PAI-1 secretion. Simvastatin may thus exhibit preventive effects against steroid-induced osteonecrosis of the femoral head by suppressing PAI-1 secretion.

Pentosan Reduces Osteonecrosis of Femoral Head in SHRSP

Increased oxidative stress is considered one of the main causes of steroid-induced osteonecrosis of the femoral head (ONFH). The aim of this study was to evaluate the effects of a steroid hormone and pentosan polysulfate sodium (pentosan), a heparin analog, in stroke-prone spontaneously hypertensive rats (SHRSP) as a model of ONFH. One hundred twenty-three 13-week-old male SHRSP//Izm rats were divided into four groups: a control group (group C), pentosan-administered group (group P), steroid-administered group (group S), and group administered pentosan plus steroid (group PS). Methylprednisolone acetate, as the steroid hormone, at a dose of 4 mg (15 mg//kg) was administered at 15 weeks of age. Pentosan at a dose of 3 mg//day//kg was continuously administered intraperitoneally from 13 weeks of age for 4 weeks. Rats were sacrificed at 17 weeks of age, and heart blood and both femora were collected. Triglyceride levels were significantly lower in group PS than in group S, indicating that pentosan improves lipid metabolism. The incidence of histologic ONFH was significantly lower in group P, at 14.8%% (10//71 femoral heads), than in group C, at 30.4%% (17//56 femoral heads), and significantly lower in group PS, at 40.8%% (29//71 femoral heads), than in group S, at 91.3%% (42//46 femoral heads), indicating that pentosan markedly inhibits ONFH. Immunohistochemical staining for oxidative stress showed that the stainability was significantly lower in group PS than in group S. Pentosan seems to reduce the incidence of ONFH in SHRSP by improving lipid metabolism and decreasing oxidative stress.

Osteonecrosis of Femoral Head in the Stroke-Prone Spontaneously Hypertensive Rats, Especially Old Rats

The average life span of stroke-prone spontaneously hypertensive rats (SHRSP) is about eight months. Male SHRSPs at 40 weeks old were used to study the idiopathic osteonecrosis of the femoral head (ION). The control group showed about 40% old necrosis and 20% early necrosis. The group administered with steroid hormone showed an increasing degeneration of adipocyte in the bone marrow, and 20% fresh necrosis was recognized. Furthermore, we observed the adipocyte change as well as early necrosis occurring among old necrosis sites. The study of aged rats may provide further understanding into the pathogenesis of ION.

Steroid changes adipokine concentration in the blood and bone marrow fluid

Our previous study has shown that plasminogen activator inhibitor 1 (PAI-1) gene expression and secretion from bone marrow adipocytes increased markedly with dexamethasone administration. The purpose of the present study was to measure the secretion of various adipokines from human bone marrow and blood, and investigate how adipokine secretion changes in a steroid environment. Human blood and bone marrow fluid were collected from a steroid treatment group and a control group during hip replacement surgery, and an enzyme-linked immunosorbent assay (ELISA) was used to measure the adiponectin, leptin, and PAI-1 levels. Adiponectin and leptin showed no significant differences between bone marrow and blood levels, but PAI-1 was significantly higher in bone marrow. The steroid treatment group had higher levels of leptin and PAI-1 in both the blood and bone marrow than the control group. PAI-1 was present at high concentrations in the bone marrow and increased by steroid treatment. High levels of PAI-1 in bone marrow may influence intraosseous hemodynamics and may induce necrotic bone disorders.

Double-bundle anterior cruciate ligament reconstruction improves tibial rotational instability: analysis of squatting motion using a 2D/3D registration technique

BackgroundThe anterior cruciate ligament-deficient (ACLD) knee requires appropriate treatment for the patient to return to sports. The purpose of this study was to clarify the kinematics of the anterior cruciate ligament-deficient knee in squatting motion before and after double-bundle anterior cruciate ligament reconstruction (DB-ACLR) using a 2D/3D registration technique.MethodsThe subjects of this study were 10 men with confirmed unilateral ACL rupture who underwent DB-ACLR. Computed tomography (CT) of the knee joints was performed before DB-ACLR. Fluoroscopic imaging of the knee motion in squatting before and after DB-ACLR was also performed. The 2D/3D registration technique is a method of calculating positional relationships by projecting the 3D bone model created from the CT data onto the image extracted from the fluoroscopic images. The tibial anteroposterior (AP) and rotational positions were analyzed with reference to the femur.ResultsThe tibial AP position of the ACLD knees was significantly anterior to the contralateral knees (p = 0.015). The tibial rotational position of the ACLD knees was significantly internally rotated compared to the contralateral knees (p < 0.001). Both tibial AP and rotational positions improved after DB-ACLR (p < 0.001), with no significant differences compared to the contralateral knees.ConclusionDB-ACLR improved not only tibial AP instability but also tibial rotational instability at knee flexion with weight-bearing. DB-ACLR appears to be a useful technique for normalizing the knee joint kinematics of ACLD knees.

Total Hip Arthroplasty Using the S-ROM-A Prosthesis for Anatomically Difficult Asian Patients

Background. The S-ROM-A prosthesis has been designed for the Asian proximal femur with a small deformed shape and narrow canal. In this study, the clinical and radiological results using the S-ROM-A prosthesis for Japanese patients with severe deformity due to dysplasia and excessive posterior pelvic tilt were examined. Methods. 94 hips were followed up for a mean of 55 months, with a mean age at surgery of 61 years. The primary diagnoses were 94 coxarthritis cases, including 51 dysplasia and 37 primary OA, 1 avascular necrosis, 2 traumatic arthritis, and 3 Perthes disease. Thirty-one hips had been treated with osteotomy of the hip joints. Preoperative intramedullary canal shapes were stovepipe in 23 hips, normal in 51 hips, and champagne-flute in 5 hips. The maximum pelvic inclination angle was 56°. Results. The mean JOA score improved from 46 points preoperatively to 80 points at final follow-up. On radiological evaluation of the fixation of the implants according to the Engh classification, 92 (97%) hips were classified as “bone ingrown fixation.” Conclusion. In primary THA, using the S-ROM-A prosthesis for Asian patients with proximal femoral deformity, even after osteotomy and with posterior pelvic tilt, provided good short- to midterm results.

Prognosis for Conservative Therapy to Vertebral Compression Fractures

胸腰椎圧迫骨折の臨床経過と予後予測について検討を行った．過去2年間で受傷早期に来院し入院治療を行った43例50椎体（男性15例，女性28例）を対象とした．3カ月以上の疼痛持続の有無とX線学的に椎体圧潰率，局所後弯を受傷時と最終観察時に計測し，受傷時MRI（T1強調像），年齢，性別，受傷時椎体圧潰率，受傷時後弯度，損傷部位，受傷機転との関連を検討した．疼痛持続，椎体圧潰進行はMRI像の後壁損傷と男性症例に関連を認めた．後弯進行はMRI像の後壁損傷に関連を認めた．胸腰椎移行部の損傷は有意に椎体圧潰が進行していた．