Yohei Naito

Anti-Tumor Necrosis Factor Therapy Inhibits Lung Metastasis in an Osteosarcoma Cell Line

Background: Osteosarcoma is the most common primary malignancy of bone, and patients often develop pulmonary metastases. In a previous study, tumor necrosis factor (TNF)-α treatment of human osteosarcoma cells increases their metastatic ability in an animal model. TNF-α can act as a tumor necrosis factor and also as a tumor-promoting factor. In the present study, the effect of a TNF-α inhibitor on osteosarcoma aggressiveness and pulmonary metastases was investigated in vitro and in vivo. Methods: The effect of infliximab, a TNF-α inhibitor, on a metastatic osteosarcoma 143B cell growth and motility was investigated in vitro. An orthotopic xenograft model of 143B cell growth and spontaneous metastasis in SCID mice was used to assess the in vivo effect of infliximab. Results: Infliximab greatly reduced cell motility and pulmonary metastases in 143B cells. The mechanism of pulmonary metastasis inhibition involved decreased expression of CXC chemokine receptor 4 (CXCR4), Rho (small GTPase protein), and its effector. Conclusions: These results suggest a novel role for TNF-α inhibition in the reduction or prevention of pulmonary metastases of osteosarcoma in this animal model. TNF-α inhibition may become a preventive therapeutic option for the pulmonary metastases of osteosarcoma.

Lumbar metastasis of choriocarcinoma.

Study Design. A case of lumbar metastasis of a choriocarcinoma is presented. Objective. To present and review a rare case of metastatic choriocarcinoma in the lumbar spine. Summary of Background Data. Choriocarcinoma is a highly anaplastic malignancy derived from trophoblastic cells characterized by the secretion of human chorionic gonadotropin (hCG) and early hematogenous metastasis. However, metastatic choriocarcinoma in the spine is extremely rare. Although 2 cases of metastasis in lumbar and/or sacral vertebra have been reported, the efficacy of surgical treatment for the spinal metastasis of choriocarcinoma is not yet known. Methods. The clinical course, radiologic features, pathology, and outcome of the treatment of metastatic choriocarcinoma of the lumbar spine is reported. Results. A 38-year-old female patient with abnormal uterine bleeding 6 weeks after a normal-term delivery showed high serum levels of hCG. A whole body image analysis revealed a lesion in the L2 vertebra. After computed tomography-guided needle biopsy, a clinical and pathologic diagnosis of lumbar metastasis of choriocarcinoma was made. Surgical resection of the localized L2 vertebra lesion was performed by total en bloc spondylectomy after a poor response to initial chemotherapy with methotrexate. Postsurgically, the serum level of hCG explosively increased and local recurrences around the original L2 vertebra and epidural metastasis abruptly developed. Lung metastases also occurred concurrently and progressed and the patient eventually died to the disease. Conclusion. We have reported a rare case of lumbar metastasis of choriocarcinoma after a normal-term pregnancy. This is the first report of lumbar metastasis of choriocarcinoma treated by spinal surgery. Because surgical resection of a lumbar metastasis of choriocarcinoma involves a substantial risk of profuse hemorrhage, local recurrence and the spread of metastasis, multiagent chemotherapy in combination with radiotherapy should be preformed before surgical resection.

A tumor endoprosthesis is useful in elderly rheumatoid arthritis patient with acute intercondylar fracture of the distal femur

The purpose of this paper is to report the use of total knee arthroplasty using a tumor prosthesis in the treatment of elderly patients with an intercondylar fracture of the distal femur. Supracondylar fractures of the femur in patients with rheumatoid arthritis are difficult to treat due to joint deformity. We present outcomes for treating intercondylar fractures of the distal femur in rheumatoid arthritis patient using a tumor endoprosthesis. This technique allows early mobilization of the patient, with restoration of a good range of knee motion. A tumor prosthesis appears to be a viable treatment option for intercondylar femoral fractures in elderly patients. It is well tolerated and permits early ambulation and return to activities of daily living.

Factors Contributing to Patient Satisfaction and Expectations following Computer-Assisted Total Knee Arthroplasty

Abstract The purpose of this study was to determine which factors contribute to patient satisfaction and expectations after total knee arthroplasty (TKA). A total of 109 patients (130 knees) with knee osteoarthritis who underwent primary TKA with navigation were studied. Intraoperative mediolateral laxity was measured. The factors including mediolateral laxity that affected patient satisfaction and expectations were evaluated using the 2011 Knee Society Score. Patient satisfaction after TKA correlated positively with symptoms and functional activities, and negatively with old age in univariate analysis. Multivariate analysis showed that symptoms and functional activities remained statistically significant; however, old age showed no difference. Patient expectations after TKA correlated positively with symptoms and functional activities, and negatively with mediolateral laxity at 60 degrees in univariate analysis. Multivariate analysis showed that symptoms, functional activities, and mediolateral laxity at 60 degrees remained statistically significant. In conclusion, midflexion instability was associated with worse expectations.

The effects of bisphosphonate on pain-related behavior and immunohistochemical analyses in hindlimb-unloaded mice

OBJECTIVE The aim of this study was to evaluate skeletal pain associated with immobility-induced osteoporosis and to examine the inhibitory effect of bisphosphonate (BP) administration on pain in hindlimb-unloaded (HU) mice. METHODS The mechanism of osteoporotic pain in HU mice was evaluated through an examination of pain-related behavior, as well as immunohistochemical findings. In addition, the effects of alendronate (ALN), a potent osteoclast inhibitor, on these parameters were assessed. RESULTS HU mice with tail suspension developed bone loss and mechanical hyperalgesia in the hindlimbs. The HU mice showed an increase in the number of calcitonin gene-related peptide (CGRP)-immunoreactive neurons and in transient receptor potential channel vanilloid subfamily member 1 (TRPV1)-immunoreactive neurons in the dorsal root ganglions (DRGs) innervating the hindlimbs. Furthermore, administration of ALN prevented HU-induced bone loss, mechanical hyperalgesia, and upregulation of CGRP and TRPV1 expressions in DRG neurons of immobility-induced osteoporotic animal models. CONCLUSIONS HU mice appear to be a useful model for immobility-induced osteoporotic pain and hindlimb-unloading-induced bone loss, as well as upregulation of CGRP and TRPV1 expressions in DRG neurons, and BP treatment prevented bone loss and mechanical hyperalgesia. The inhibitory effect of BP on osteoclast function might contribute to improving osteoporosis-related pain.

FRI0566 Teriparatide and alendronate improved bone loss and hyperalgesia in a mouse model of osteoporosis

Background Osteoporosis may cause not only fractures but also chronic back pain in elderly women. Teriparatide (TPTD) and alendronate (ALN) are widely used in clinical for treatment of osteoporosis. Several studies have demonstrated that TPTD and ALN treatments improved skeletal pain in osteoporosis patients. Objectives We investigated the effect of TPTD and ALN on pain-related behavior in ovariectomized (OVX) mice. And we investigated expression of inflammatory cytokines treated OVX mice. Methods 8-week-old female ddY mice were OVX and assigned to 4groups; SHAM-operated mice treated with vehicle (SHAM), OVX mice treated with vehicle (OVX), OVX mice treated with TPTD (TPTD) and OVX mice treated with ALN (ALN). Mice were started treatment immediately after surgery. For 4 weeks, mice were injected subcutaneously with vehicle or 40μg/kg ALN twice a week or 40μg/kg TPTD 5 times a week. The bilateral distal femur metaphyses were analyzed three-dimensionally by μCT 4 weeks after surgery (each group; n=8). Mechanical sensitivity was tested using von Frey filaments 4 weeks after surgery. To evaluate the 50% withdrawal threshold, seven von Frey filaments with forces of 0.07, 0.16, 0.4, 0.6, 1.0, 1.4 and 2.0 g were applied to the middle of the plantar surface. Data was collected using the up-down method. To evaluate expression of interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α), mice were anesthetized and the bilateral hindlimb bone excised. We performed quantitative polymerase chain reaction (q-PCR) from hindlimb bone. Results μCT analysis of the distal femur metaphysis showed that bone volume/tissue volume (BV/TV) and trabecular number (Tb.N) were significantly less in the OVX group than in the SHAM group, whereas trabecular separation (Tb.Sp) was significantly greater in the OVX group than in the SHAM group. In the TPTD and ALN group, BV/TV and Tb.N were significantly greater than in the OVX group, whereas Tb.Sp was significantly less than in the OVX group. And in the ALN group, BV/TV and Tb.N were significantly greater than in the TPTD group, but Tb.Sp was no significance. The 50% withdrawal threshold was significantly lower in the OVX group than in the SHAM group, and it was significantly higher in the TPTD and ALN group than in the OVX group. And the 50% withdrawal threshold was no significance between the TPTD and ALN group. The expression levels of TNF-α was increased in the OVX group compared with those in the SHAM group. Other cytokines were not increased significantly in the OVX group. In the TPTD and ALN group, the expression levels of TNF-α was significantly degreased than the OVX group. And the expression levels of TNF-α was no significance between the TPTD and ALN group. Conclusions In this study, TPTD and ALN treatments prevented bone loss in OVX mice. Mechanical hyperalgesia in hindlimbs tended to be decreased in the OVX group compared with the TPTD and ALN group. ALN treatment was more effective in bone formation compared with TPTD treatment, whereas pain relief was no significance between TPTD and ALN treatment. These results suggest that TPTD treatment was more effective in osteoporosis patients with skeletal pain. Disclosure of Interest None declared

THU0487 Teriparatide Improved Bone Loss and Hyperalgesia in A Mouse Model of Osteoporosis

Background Osteoporosis may cause not only fractures but also chronic back pain in elderly women. Bente reported that postmenopausal women with severe osteoporosis who were prescribed teriparatide (TPTD) in standard clinical practice had a reduction in back pain. However, there have been no reports regarding the effect of TPTD on pain in animal models of osteoporosis. Objectives The objective of the current study was to investigate the effect of TPTD on pain-related behavior in ovariectomized (OVX) mice. Methods Female ddY mice (8 weeks old) were ovariectomized and assigned to 3groups; SHAM-operated mice treated with vehicle (SHAM), OVX mice treated with vehicle (OVX), OVX mice treated with TPTD (TPTD). Starting immediately after surgery, vehicle or 40μg/kg TPTD was injected subcutaneously 5 times a week for 4 weeks. The proximal tibial metaphysis were analyzed three-dimensionally by μCT 4 weeks after surgery (each group; 8 mice). Mechanical sensitivity was tested using von Frey filaments 4 weeks after surgery. To evaluate the withdrawal threshold, each von Frey filaments was applied once, starting with 0.008g and increasing until a withdrawal response was reached, which was considered a positive response. The lowest force producing a response was considered the withdrawal threshold. To evaluate the 50% withdrawal threshold, seven von Frey filaments were applied to the middle of the plantar surface. Testing was initiated with the 0.6g filament. In the absence of a clear paw withdrawal response, increasingly stronger filaments were presented consecutively, until one of them was found to elicit such a response. If the 0.6 g filament elicited a response, filaments with decreasing strength were presented until determination of the first one which failed to cause paw withdrawal. Data was collected using the up-down method. To identify osteoclasts in hindlimb bone, we used the tartrate-resistant acid phosphatase (TRAP) method. TRAP-positive multinucleated cells were scored as osteoclasts. Measurements were made within an area the proximal tibial metaphysis starting immediately below the growth plate. Histomorphometry was conducted to quantify the number of osteoclasts. Results μCT analysis of the proximal tibial metaphysis showed that bone volume/tissue volume (BV/TV) and trabecular number (Tb.N) were significantly less in the OVX group than in the SHAM group, whereas trabecular separation (Tb.Sp) was significantly greater in the OVX group than in the SHAM group. In the TPTD group, BV/TV and Tb.N were significantly greater than in the OVX group, whereas Tb.Sp was significantly less than in the OVX group. The withdrawal threshold was significantly lower in the OVX group than in the SHAM group, and it was significantly higher in the TPTD group than in the OVX group. Similarly, the 50% withdrawal threshold was significantly lower in the OVX group than in the SHAM group, and it was significantly higher in the TPTD group than in the OVX group. The number of osteoclasts was significantly higher in the OVX group than in the SHAM group, whereas it was higher in the TPTD group than in the OVX group, but there were no significantly changes. Conclusions TPTD prevented ovariectomy-induced bone loss. In addition, mechanical hyperalgesia in hindlimbs significantly improved in OVX group compared with TPTD group. The results suggest that TPTD prevented postmenopausal osteoporotic pain. Disclosure of Interest None declared

AB0781 Effects of Teriparatide for Bone Loss and Pain-Related Behavior in The Hind Limb-Unloaded Mouse Model of Disuse Osteoporosis

Background Factors of chronic pain due to hemiplegia or prolonged immobility are varied, there is osteoporosis (a rarefaction due to bone atrophy) as the cause. Osteoporotic patients with no evidence of fractures sometimes experience vague lower back pain. However, there have been few reports regarding the correlation between osteoporosis and pain-related behavior. Our previous studies have indicated that hindlimb-unloading induced bone loss and mechanical hyperalgesia in hindlimb of mice. We investigated the effects of teriparatide (PTH) on bone mass and pain-related behaviors in osteoporosis model mice with hindlimb unloading. Objectives The objective of the current study was to investigate the effect of PTH on bone mass and pain-related behaviors in hindlimb-unloaded mice model of disuse osteoporosis. Methods Male ddY mice (8 weeks old) were tail-suspended for 2 weeks and assigned to 3 groups; hindlimb-loaded mice treated with vehicle (control), hindlimb-unloaded mice treated with vehicle (HU), hindlimb-unloaded mice treated with PTH (HU-PTH) (n=8/group). Starting immediately after tail-suspension, vehicle or 40μg/kg PTH was injected subcutaneously twice a week for 2 weeks. The bilateral distal femoral metaphyses and proximal tibial metaphyses were analyzed three-dimensionally by micro-computed tomography (μCT) 2 weeks after tail-suspension. Mechanical sensitivity was also tested using von Frey filaments 2 week after the tail suspension. The withdrawal threshold, the 50% withdrawal threshold and the frequency of the withdrawal response to the application of von Frey filaments to the plantar surface of the hind paws was examined. Measurement of pain-related behavior with von Frey filaments was interpreted as indicative of mechanical allodynia. Results μCT analysis of the distal femoral metaphysis and the proximal tibial metaphysis showed that significantly decreased bone volume/tissue volume (BV/TV) and trabecular thickness (Tb.Th) in HU compared with HL was significantly increased by PTH treatment. (Fig.1) Similarly, decreased trabecular number (Tb.N) in HU compared with HL was increased, and increased trabecular separation (Tb.Sp) in HU compared with HL was decreased by PTH treatment, but not significant. The paw withdrawal threshold and the 50% paw withdrawal threshold were significantly lower in the HU than in the control, whereas it was significantly higher in the HU-PTH than in the HU group. (Fig.2) The paw withdrawal frequency stimulated by von Frey filaments with strength of 0.4–2.0 g was significantly higher in the HU than in the control. Whereas it was significantly lower in the HU-PTH than in the HU. Conclusions In this study, treatment of PTH prevented bone loss and mechanical hyperalgesia in disuse osteoporotic animal models by hindlimb-unloading. The results suggest that low bone volume itself is one of the causes of osteoporotic pain. Disclosure of Interest None declared

FRI0306 Effects of Bisphosphonate for Pain-Related Behavior and Immunohistochemical Analysis in the Hind Limb-Unloaded Mouse Model of Disuse Osteoporosis

Background Osteoporotic patients with no evidence of fractures sometimes experience vague lower back pain. A previous report has indicated that sensory innervation of ovariectomized rat vertebrae showed increased expression of the CGRP, a neuropeptide marker of pain, and the TRPV1, an acid-sensitive ion channel, in dorsal root ganglion (DRG) neurons1. However, there have been few reports regarding the correlation between osteoporosis, pain-related behavior and immunohistochemical analysis. Objectives The objective of the current study was to investigate pain-related behavior and to elucidate the mechanism of osteoporotic pain by immunohistochemical analysis of the DRG in hindlimb-unloaded (HU) mice model of disuse osteoporosis2. And we investigated the effect of alendronate (ALN) in HU mice. Methods Male ddY mice (8 weeks old) were tail-suspended for 2 week and assigned to 3 groups; hindlimb-loaded mice treated with vehicle (control), HU mice treated with vehicle (HUV), HU mice treated with ALN (HUA)(n=8/group). Starting immediately after tail-suspension, vehicle or 40μg/kg ALN was injected subcutaneously twice a week for 2 weeks. The bilateral distal femoral metaphyses and proximal tibial metaphyses were analyzed three-dimensionally by μCT 2 weeks after tail-suspension. Mechanical sensitivity was also tested using von Frey filaments 2 week after the tail suspension. The withdrawal threshold, the 50% withdrawal threshold and the frequency of the withdrawal response to the application of von Frey filaments to the plantar surface of the hind paws was examined. Immunohistochemical analysis of the CGRP expression and the TRPV1 expression was completed for the L4 and L5 DRG neurons. Results μCT analysis of the distal femoral metaphysis (Fig. 1a) and the proximal tibial metaphysis showed that significantly decreased bone volume/tissue volume (BV/TV) and trabecular number (Tb.N) in HUV compared with control was significantly increased by ALN treatment. Significantly increased trabecular separation (Tb.Sp) in HUV compared with control was significantly decreased by ALN treatment. The paw withdrawal threshold and the 50% paw withdrawal threshold were significantly lower in the HUV than in the control, whereas it was significantly higher in the HUA than in the HUV group (Fig. 1b). The paw withdrawal frequency stimulated by von Frey filaments with strength of 0.4–2.0 g was significantly higher in the HUV than in the control. Whereas it was significantly lower in the HUA than in the HUV. The immunohistochemical analysis showed that the ratio of CGRP-immunoreactive (ir) and TRPV1-ir DRG neurons (L4 and L5) in the HUV was significantly higher than in the control, whereas it was significantly lower in the HUA than in the HUV. There was no significant difference between the HUA and the control in μCT analysis, hyperalgesia of hindlimbs and immunohistochemical analysis. Conclusions In this study, treatment of ALN prevented bone loss, mechanical hyperalgesia and upregulation of CGRP and TRPV1 expression in DRG neurons of disuse osteoporotic animals models by hindlimb-unloading. The results suggest that bone resorption is one of the causes of osteoporotic pain. References Orita S et al. Spine. 2010. Barou O et al. Invest Radiol. 2002. Disclosure of Interest None declared

SAT0300 Teriparatide Inhibits Bone Loss and Improves Hyperalgesia in Ovariectomized Mice

Background Osteoporosis may cause not only fractures but also chronic back pain in elderly women. Bente et al reported that postmenopausal women with severe osteoporosis who were prescribed teriparatide (TPTD) in standard clinical practice had a reduction in back pain. However, there have been no reports regarding the effect of TPTD on pain in animal models of osteoporosis. Objectives In this study, we investigated the effect of TPTD on pain-related behavior in ovariectomized mice. Methods 8-week-old female ddY mice were ovariectomized (OVX) and assigned to 3groups; SHAM-operated mice treated with vehicle (SHAM), OVX mice treated with vehicle (OVX-V), OVX mice treated with TPTD (OVX-TPTD). Starting immediately after surgery, vehicle or 40μg/kg TPTD was injected subcutaneously 5 times a week for 4 weeks. The bilateral proximal tibial metaphyses were analyzed three-dimensionally by μCT 4 weeks after surgery (each group; 4 mice). Mechanical sensitivity was tested using von Frey filaments 4 weeks after surgery. To evaluate the 50% withdrawal threshold, nine von Frey filaments with forces of 0.02, 0.04, 0.07, 0.16, 0.4, 0.6, 1.0, 1.4 and 2.0 g were applied to the middle of the plantar surface. Testing was initiated with the 0.6 g filament. In the absence of a clear paw withdrawal response, increasingly stronger filaments were presented consecutively, until one of them was found to elicit such a response. If the 0.6 g filament elicited a response, filaments with decreasing strength were presented until determination of the first one which failed to cause paw withdrawal. Data was collected using the up-down method to calculate the 50% mechanical paw withdrawal threshold Results μCT analysis of the proximal tibial metaphysis (Fig. 1) showed that bone volume/tissue volume (BV/TV) and trabecular number (Tb.N) were significantly less in the OVX-V group than in the SHAM group, whereas trabecular separation (Tb.Sp) was significantly greater in the OVX-V group than in the SHAM group. In the OVX-TPTD group, BV/TV and Tb.N were significantly greater than in the OVX-V group, whereas Tb.Sp was significantly less than in the OVX-V group. The 50% withdrawal threshold was significantly lower in the OVX group than in the SHAM group, whereas it was higher in the OVX-TPTD group than in the OVX-V group,but there was not significant. Conclusions In this study,TPTD prevented ovariectomy-induced bone loss. Mechanical hyperalgesia in hindlimbs tended to be decreased in OVX group compered with OVX-TPTD group.TPTD may be usefull for treatment of the postmenopausal osteoporotic pain. Disclosure of Interest None declared