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Biomedical subjects

Hideki Yoshikawa

Publications and source records attributed to Hideki Yoshikawa.

At least 73 records · Page 4Linked to original sources
PubMed2005

Differential and collaborative actions of Rad51 paralog proteins in cellular response to DNA damage.

Metazoan Rad51 plays a central role in homologous DNA recombination, and its activity is controlled by a number of Rad51 cofactors. These include five Rad51 paralogs, Rad51B, Rad51C, Rad51D, XRCC2 and XRCC3. We previously hypothesized that all five paralogs participate collaboratively in repair. However, this idea was challenged by the biochemical identification of two independent complexes composed of either Rad51B/C/D/XRCC2 or Rad51C/XRCC3. To investigate if this biochemical finding is matched by genetic interactions, we made double mutants in either the same complex (rad51b/rad51d) or in both complexes (xrcc3/rad51d). In agreement with the biochemical findings the double deletion involving both complexes had an additive effect on the sensitivity to camptothecin and cisplatin. The double deletion of genes in the same complex, on the other hand, did not further increase the sensitivity to these agents. Conversely, all mutants tested displayed comparatively mild sensitivity to gamma-irradiation and attenuated gamma-irradiation-induced Rad51 foci formation. Thus, in accord with our previous conclusion, all paralogs appear to collaboratively facilitate Rad51 action. In conclusion, our detailed genetic study reveals a complex interplay between the five Rad51 paralogs and suggests that some of the Rad51 paralogs can separately operate in later step of homologous recombination.

Animals↗
PubMed2005

Leptin regulates chondrocyte differentiation and matrix maturation during endochondral ossification.

Leptin has been suggested to mediate a variety of actions, including bone development, via its ubiquitously expressed receptor (Ob-Rb). In this study, we investigated the role of leptin in endochondral ossification at the growth plate. The growth plates of wild-type and ob/ob mice were analyzed. Effects of leptin on chondrocyte gene expression, cell cycle, apoptosis and matrix mineralization were assessed using primary chondrocyte culture and the ATDC5 cell differentiation culture system. Immunohistochemistry and in situ hybridization showed that leptin was localized in prehypertrophic chondrocytes in normal mice and that Ob-Rb was localized in hypertrophic chondrocytes in normal and ob/ob mice. Growth plates of ob/ob mice were more fragile than those of wild-type mice in a mechanical test and were broken easily at the chondro-osseous junction. The growth plates of ob/ob mice showed disturbed columnar structure, decreased type X collagen expression, less organized collagen fibril arrangement, increased apoptosis and premature mineralization. Leptin administration in ob/ob mice led to an increase in femoral and humeral lengths and decrease in the proportional length of the calcified hypertrophic zone to the whole hypertrophic zone. In primary chondrocyte culture, the matrix mineralization in ob/ob chondrocytes was stronger than that of wild-type mice; this mineralization in both types of mice was abolished by the addition of exogenous leptin (10 ng/ml). During ATDC5 cell differentiation culture, exogenous leptin at a concentration of 1-10 ng/ml (equivalent to the normal serum concentration of leptin) altered type X collagen mRNA expression and suppressed apoptosis, cell growth and matrix calcification. In conclusion, we demonstrated that leptin modulates several events associated with terminal differentiation of chondrocytes. Our finding that the growth plates of ob/ob mice were fragile implies a disturbance in the differentiation/maturation process of growth plates due to depletion of leptin signaling in ob/ob mice. These findings suggest that peripheral leptin signaling plays an essential role in endochondral ossification at the growth plate.

Animals↗
PubMed2005

Adiponectin increases bone mass by suppressing osteoclast and activating osteoblast.

Adiponectin, an adipose-derived hormone, exhibits various biological functions, such as increasing insulin sensitivity, protecting hypertension, and suppression of atherosclerosis, liver fibrosis, and tumor growth. Here, we report the role of adiponectin on bone metabolism. C57BL/6J mice were treated with adenovirus expressing lacZ or adiponectin, and their bones were analyzed by three-dimensional microcomputed tomography. Adiponectin-adenovirus treatment increased trabecular bone mass, accompanied by decreased number of osteoclasts and levels of plasma NTx, a bone-resorption marker. In vitro studies showed that adiponectin inhibited M-CSF- and RANKL-induced differentiation of mouse bone marrow macrophages and human CD14-positive mononuclear cells into osteoclasts and also suppressed the bone-resorption activity of osteoclasts. Furthermore, adiponectin enhanced mRNA expression of alkaline phosphatase and mineralization activity of MC3T3-E1 osteoblasts. Our results indicate that adiponectin exerts an activity to increase bone mass by suppressing osteoclastogenesis and by activating osteoblastogenesis, suggesting that adiponectin manipulation could be therapeutically beneficial for patients with osteopenia.

Adenoviridae↗
PubMed2005

Macro-structural effect of metal surfaces treated using computer-assisted yttrium-aluminum-garnet laser scanning on bone-implant fixation.

Porous coatings have been applied to the surface of prosthetic devices to foster stable device fixation. The coating serves as a source of mechanical interlocking and may stimulate healthy bone growth through osseointegrated load transfer in cementless arthroplasty. Joint arthroplasty by porous-coated prostheses is one of the most common surgical treatments, and has provided painless and successful joint mobility. However, long-term success is often impaired by the loss of fixation between the prosthesis and bone. Porous-coated prostheses are associated with several disadvantages, including metal debris from porous coatings (third body wear particles) and irregular micro-texture of metal surfaces. Consequently, quantitative histological analysis has been very difficult. These issues arise because the porous coating treatment is based on addition of material and is not precisely controllable. We recently developed a precisely controllable porous texture technique based on material removal by yttrium-aluminum-garnet laser. Free shapes can be applied to complex, three-dimensional hard metal surfaces using this technique. In this study, tartan check shapes made by crossing grooves and dot shapes made by forming holes were produced on titanium (Ti6A14V) or cobalt chrome (CoCr) and evaluated with computer-assisted histological analysis and measurement of bone-metal interface shear strength. Width of grooves or holes ranged from 100 to 800 mum (100, 200, 500, and 800 microm), with a depth of 500 microm. When the cylindrical porous-texture-treated metal samples (diameter, 5 mm; height, 15 mm) were implanted into a rabbit femoral condyle, bone tissue with bone trabeculae formed in the grooves and holes after 2 or 4 weeks, especially in 500-microm-wide grooves. Abundant osteoconduction was consistently observed throughout 500-microm-wide grooves in both Ti6A14V and CoCr. Speed of osteoconduction was faster in Ti6A14V than in CoCr, especially in the tartan check shape made of 500-microm-wide grooves. In pushout testing, the tartan check shape made of 500-microm-wide grooves had significantly higher bone-metal interface shear strength than the dot shape or commercial porous coating. These results indicate that the tartan check shape made of 500-microm-wide grooves on metal surfaces has potential for clinical application in artificial prosthesis design.

Aluminum↗
PubMed2005

Apoptosis induced by the sonomechanical effects of low intensity pulsed ultrasound in a human leukemia cell line.

To obtain an optimal condition for ultrasound (US)-induced apoptosis that could be useful for cancer therapy, we applied low intensity pulsed US to sonicate U937 cells in vitro. Cells were then incubated at different time intervals before measuring apoptosis. The apoptosis was assessed by DNA fragmentation and phosphatidylserine externalization. The pattern of the decrease in mitochondrial membrane potential was determined by flow cytometry. Optimal apoptosis (70.0+/-13.8%) with minimal lysis was attained with 1 MHz ultrasound 0.3 W/cm2, 10% duty factor at 100 Hz for 1 min) at 12 h after sonication. Lack of US-induced free radical detection and absence of Heme oxygenase-1, an intracellular oxidative stress marker, up-regulation in cells, suggest that sonomechanical, not sonochemical, effects are the main mechanism involved.

Apoptosis↗
PubMed2005

FDG-PET imaging for chronic expanding hematoma in pelvis with massive bone destruction.

Chronic expanding hematoma is a rare presentation of a hematoma characterized by a persistent increase in size for more than a month after the initial hemorrhage. We present a 65-year-old man with a chronic expanding hematoma in his ilium who was receiving anticoagulant treatment. The patient had a delayed manifestation of a femoral neuropathy with massive bone destruction. 2-Deoxy-[18F]fluoro-D: -glucose (FDG) positron emission tomography (PET) imaging revealed an increased uptake in the rim of the mass in images acquired 1 h after FDG injection. FDG-PET scans were performed using a dedicated PET scanner (HeadtomeV/SET2400 W, Shimadzu, Kyoto, Japan), and the PET data for the most metabolically active region of interest (ROI) were analyzed. The maximum standardized uptake value (SUVmax) was set to a cut-off point of 3.0 to distinguish between benign and malignant lesions. The SUVmax of the patient's lesion was 3.10, suggesting a malignant lesion. The characteristics of FDG-PET images of chronic expanding hematomas, including the uptake of FDG in the peripheral rim of the mass as a result of inflammation, should be recognized as a potential interpretive pitfall in mimicking a sarcoma.

Aged↗
PubMed2005

Long-term outcome of laminoplasty for cervical myelopathy due to disc herniation: a comparative study of laminoplasty and anterior spinal fusion.

STUDY DESIGN: A retrospective study was conducted. OBJECTIVE: To compare the long-term outcomes after laminoplasty and anterior spinal fusion (ASF) for cervical myelopathy secondary to disc herniation. SUMMARY OF BACKGROUND DATA: There have been no reports of long-term comparative studies of laminoplasty and ASF for cervical myelopathy due to disc herniation. METHODS: Of 21 patients who underwent ASF only between 1984 and 1987, 15 were followed up. Of 22 patients who underwent laminoplasty only between 1987 and 1994, 18 were followed up. There were no significant differences in preoperative prognostic factors between the 2 groups. Average follow-up was 15 years in the ASF group and 10 years in the laminoplasty group. Neurologic and radiologic results were examined. RESULTS: Laminoplasty and ASF provided equal neurologic improvement. In the ASF group, additional surgery was required for bone graft complications in 2 patients and for adjacent spondylosis in 1. In the laminoplasty group, one patient had C5 palsy, and intractable axial pain developed in 5 patients after surgery, but no patients needed additional surgery. CONCLUSIONS: Because the 2 procedures provided the same neurologic improvement, the risks of bone graft complication with ASF must be weighed against the risks of chronic neck pain associated with laminoplasty for determining the best technique. Therefore, because our present surgical strategy for cervical myelopathy due to disc herniation, laminoplasty is the procedure of choice except for a patient with single level disc herniation without developmental canal stenosis, who is considered to be a good candidate for ASF.

Bone Transplantation↗
PubMed2005

Assessment of the three-dimensional relationship of the ossific nuclei and cartilaginous anlagen in congenital clubfoot by 3-D MRI.

PURPOSE: Radiographic measurement is the usual method used to objectively determine the extent of a congenital clubfoot deformity. Although radiographs have been used clinically to estimate the size and location of tarsal bones through measurements of the ossific nuclei, it is not clear to what extent these relationships are actually reflected in these measurements. So, we used a 3-D MRI system that could more objectively estimate sizes and positional relationships. MATERIAL AND METHOD: We evaluated 5 patients with unilateral congenital clubfoot deformity. Magnetic resonance imaging was performed of both feet using 1.5-T magnet. Based on the resulting magnetic resonance imaging volume data, a three-dimensional surface bone model was reconstructed by the Marching Cubes method. We used this model to perform a comparative analysis of the volume and volume ratio of each cartilaginous anlage and ossific nucleus, the length of the talus and the calcaneus, and the position of the center of gravity of ossific nuclei within the cartilaginous anlagen. We measured the relationship between the ossific nuclei and cartilaginous anlagen in the talus and calcaneus of patients with unilateral clubfoot deformity. RESULT: In clubfeet talus volume was reduced by 20.1% and calcaneal volume was reduced by 15.7%. Furthermore, the volume of the talar ossific nucleus was reduced by 42.6% and that of the calcaneal ossific nucleus was reduced by 12.1%. The length of the clubfoot talus was 8.2% shorter than normal, and that of the calcaneus was 4.8% shorter. CONCLUSION: The assessment technique presented herein was shown to be useful in ascertaining the various pathological characteristics associated with clubfoot.

Calcaneus↗
PubMed2005

[Development of three-dimensional kinematic analysis system for artificial knee implants using X-ray fluoroscopic imaging].

To achieve quantitative assessment of 3D dynamic motion of artificial knee implants under clinical conditions, we developed a 3D kinematic analysis system using X-ray fluoroscopic imaging. The 3D pose-estimation technique for knee implants was built on a 2D/3D registration algorithm, which determines the spatial pose for each femoral and tibial component from the knee implant contours and computer-assisted design (CAD) models of the implant. In order to validate the accuracy of the 3D pose estimation and the system, computer simulation and in vitro tests were performed using images of knee implants taken in 10 different poses with respect to X-ray focus. Computer simulation tests showed that the root mean square errors (RMSE) for all variables were less than 1.0 mm 1.0 degrees. In vitro tests showed that the RMSE for translation perpendicular to the X-ray image plane was about 1.5 mm, while the accuracy of the remaining two translational and three rotational variables was found to be sufficient for analyzing knee kinematics. Computation time in 3D pose estimation was then obtained in less than 30 seconds for each frame. In clinical application, dynamic movement in deep knee bending was quantitatively analyzed, and the feasibility and effectiveness of the system was demonstrated.

Algorithms↗
PubMed2005

Microbubble-enhanced ultrasound exposure promotes uptake of methotrexate into synovial cells and enhanced antiinflammatory effects in the knees of rabbits with antigen-induced arthritis.

OBJECTIVE: To evaluate whether microbubble-enhanced ultrasound (US) treatment promotes the delivery of methotrexate (MTX) into synovial cells and the enhanced antiinflammatory effects of intraarticular MTX therapy in a rabbit arthritis model. METHODS: Arthritis was induced in both knees of 53 rabbits by immunization with ovalbumin. MTX including a microbubble agent was then injected into the left and right knee joints, and the right knees were exposed to US (MTX+/US+ group), while the left knees were not (MTX+/US- group). The knee joints were evaluated histologically in 7 rabbits at 5 time points up to day 56. Quantitative gene expression of interleukin-1beta (IL-1beta) in synovial tissue was measured on days 7 and 28. Eight rabbits were used for the measurement of MTX concentration in synovial tissue 12 hours after treatment. To evaluate the effect of microbubble-enhanced US treatment in the absence of MTX, only the microbubble agent was injected into the left and right knee joints of 10 rabbits with or without US exposure, and these animals were evaluated histologically on days 7 and 28. RESULTS: The MTX concentration in synovial tissue was significantly higher in the MTX+/US+ group than in the MTX+/US- group. Synovial inflammation was less prominent in the MTX+/US+ group compared with the MTX+/US- group, judging from the results of the histologic evaluation and the gene expression levels of IL-1beta in synovial tissue. It also appeared that microbubble-enhanced US exposure itself did not affect inflammation. CONCLUSION: Microbubble-enhanced US exposure promoted the uptake of MTX into synovial cells, which resulted in enhancement of the antiinflammatory effects of the intraarticular MTX injection. These results suggest that application of this technique may have clinical benefit.

Animals↗
PubMed2005

Nurse-like cells from patients with rheumatoid arthritis support the survival of osteoclast precursors via macrophage colony-stimulating factor production.

OBJECTIVE: To elucidate the role of nurse-like cells (NLCs) obtained from rheumatoid arthritis (RA) patients in bone loss during progressive synovial expansion. METHODS: CD14+ monocytes were cocultured with NLCs for 4 weeks and collected as NLC-supported CD14+ (NCD14+) monocytes. To determine their ability to differentiate into osteoclasts, NCD14+ monocytes were further cultured with macrophage colony-stimulating factor (M-CSF) together with RANKL or tumor necrosis factor alpha (TNFalpha). NCD14+ monocytes were also cocultured with SaOS-4/3 cells, which were shown to support osteoclastogenesis in response to parathyroid hormone (PTH). CD14+ monocytes were cocultured with SaOS-4/3 cells to elucidate how SaOS-4/3 cells and NLCs supported CD14+ monocytes for a long period. Synovial expansion adjacent to bone in RA patients was examined immunohistochemically to detect osteoclast precursors such as NCD14+ monocytes. RESULTS: NLCs supported the survival of CD14+ monocytes for 4 weeks. NCD14+ as well as CD14+ monocytes differentiated into osteoclasts in the presence of M-CSF together with RANKL or TNFalpha. NCD14+ monocytes also differentiated into osteoclasts in PTH-treated cocultures with SaOS-4/3 cells. SaOS-4/3 cells supported the survival of CD14+ monocytes for 4 weeks in the presence, but not absence, of PTH. Treatment of SaOS-4/3 cells with PTH up-regulated the expression of M-CSF messenger RNA. Neutralizing antibodies against M-CSF inhibited the NLC-supported survival of CD14+ monocytes. CD68+ monocytes and M-CSF+ fibroblast-like synoviocytes were colocalized in regions adjacent to the destroyed bone of RA patients. CONCLUSION: Our findings suggest that NLCs are involved in RA-induced bone destruction by maintaining osteoclast precursors via production of M-CSF.

Aged↗
PubMed2005

Application of 3D-MR image registration to monitor diseases around the knee joint.

PURPOSE: To estimate the accuracy and consistency of a method using a voxel-based MR image registration algorithm for precise monitoring of knee joint diseases. MATERIALS AND METHODS: Rigid body transformation was calculated using a normalized cross-correlation (NCC) algorithm involving simple manual segmentation of the bone region based on its anatomical features. The accuracy of registration was evaluated using four phantoms, followed by a consistency test using MR data from the 11 patients with knee joint disease. RESULTS: The registration accuracy in the phantom experiment was 0.49+/-0.19 mm (SD) for the femur and 0.56+/-0.21 mm (SD) for the tibia. The consistency value in the experiment using clinical data was 0.69+/-0.25 mm (SD) for the femur and 0.77+/-0.37 mm (SD) for the tibia. These values were all smaller than a voxel (1.25 x 1.25 x 1.5 mm). CONCLUSION: The present method based on an NCC algorithm can be used to register serial MR images of the knee joint with error on the order of a sub-voxel. This method would be useful for precisely assessing therapeutic response and monitoring knee joint diseases; normalized cross-correlation; accuracy.

Algorithms↗
PubMed2005

Bone tissue engineering with porous hydroxyapatite ceramics.

The main principle of bone tissue engineering strategy is to use an osteoconductive porous scaffold in combination with osteoinductive molecules or osteogenic cells. The requirements for a scaffold in bone regeneration are: (1) biocompatibility, (2) osteoconductivity, (3) interconnected porous structure, (4) appropriate mechanical strength, and (5) biodegradability. We recently developed a fully interconnected porous hydroxyapatite (IP-CHA) by adopting the "form-gel" technique. IP-CHA has a three-dimensional structure with spherical pores of uniform size that are interconnected by window-like holes; the material also demonstrated adequate compression strength. In animal experiments, IP-CHA showed superior osteoconduction, with the majority of pores filled with newly formed bone. The interconnected porous structure facilitates bone tissue engineering by allowing the introduction of bone cells, osteotropic agents, or vasculature into the pores. In this article, we review the accumulated data on bone tissue engineering using the novel scaffold, focusing especially on new techniques in combination with bone morphogenetic protein (BMP) or mesenchymal stem cells.

Animals↗
PubMed2005

Bone metastasis of intracranial meningeal hemangiopericytoma.

The authors investigated the clinical and radiological features and the treatment strategy for bone metastasis from a rare tumor, intracranial meningeal hemangiopericytoma. The clinical presentations and characteristic imagings were retrospectively reviewed in 15 bony metastatic lesions of four patients with this tumor. All four cases were initially diagnosed as atypical meningioma, and all of the bone metastases developed more than 10 years after the initial intracranial surgery. The common symptom induced by the metastatic lesions was pain. On plain roentgen films and computed tomography (CT), the involved bones showed thinning and expansion. On bone scintigraphy, a "cold-in-hot" appearance was typically observed. The clinical and radiological findings were diagnosed as bone metastasis from intracranial meningeal hemangiopericytoma. The prognosis after bone metastasis was relatively short compared to the long duration before bone metastases, because of the coexistence of other, visceral, metastasis. Combined with effective radiation therapy, surgical intervention for bone metastasis of this tumor should be carefully considered.

Aged↗
PubMed2005

Graft length changes in the bi-socket anterior cruciate ligament reconstruction: comparison between isometric and anatomic femoral tunnel placement.

PURPOSE: To compare the length change of grafts in bi-socket anterior cruciate ligament (ACL) reconstruction between isometric and anatomic placement. TYPE OF STUDY: Case series. METHODS: The subjects were 110 knees of 110 patients with a mean age of 27 years. All knees were reconstructed using multistranded autogenous semitendinosus tendon and the bi-socket procedure. Of those, 43 knees were reconstructed in the isometric position, around the isometric point (group I) and the remaining 67 in the anatomic position, mimicking the anatomic footprint of ACL (group II). Grouping was performed based on the femoral socket position. Each femoral socket for the anterior medial graft (AMG) and the posterolateral graft (PLG) for the left knee was positioned at 1 o'clock and 2 o'clock to 2:30 in group I, or 2 o'clock and 3 o'clock in group II in the posterior border of the notch. Their positioning for the right knee was at 11 o'clock and 9:30 to 10 o'clock in group I, or 10 o'clock and 9 o'clock in group II. The length changes of grafts were intraoperatively measured while extending the knee from 120 degrees to full extension, using a commercially available isometric positioner. The Student t test was used for statistical analysis. RESULTS: All the grafts showed the pattern of elongation approaching extension. The length change of AMG and PLG was 1.3+/-0.6 mm and 2.2+/-1.0 mm in the group I, 1.4+/-0.7 mm and 2.5+/-0.9 mm in the group II, respectively. There was no statistically significant difference in the values of AMG (P = .51) and PLG (P = .074) between the 2 groups. CONCLUSIONS: The length change of ACL grafts placed in the anatomic position was not significantly different from that positioned in the isometric position. LEVEL OF EVIDENCE: Level IV, case series.

Adolescent↗
PubMed2005

Potentiation of the activity of bone morphogenetic protein-2 in bone regeneration by a PLA-PEG/hydroxyapatite composite.

Bone morphogenetic proteins (BMPs) are biologically active molecules capable of inducing new bone formation, and show potential for clinical use in bone defect repair. However, an ideal system for delivering BMPs that can potentiate their bone-inducing ability and provide initial mechanical strength and scaffold for bone ingrowth has not yet been developed. In this study, to construct a carrier/scaffold system for BMPs, we combined two biomaterials: interconnected-porous calcium hydroxyapatite ceramics (IP-CHA), and the synthetic biodegradable polymer poly D,L,-lactic acid-polyethyleneglycol block co-polymer (PLA-PEG). We used a rabbit radii model to evaluate the bone-regenerating efficacy of rhBMP-2/PLA-PEG/IP-CHA composite. At 8 weeks after implantation, all bone defects in groups treated with 5 or 20 microg of BMP were completely repaired with sufficient strength. Furthermore, using this carrier scaffold system, we reduced the amount of BMP necessary for such results to about a tenth of the amount needed in previous studies, probably due to the superior osteoconduction ability of IP-CHA and the optimal drug delivery system provided by PLA-PEG, inducing new bone formation in the interconnected pores. The present findings indicate that the synthetic biodegradable polymer/IP-CHA composite is an excellent combination carrier/scaffold delivery system for rhBMP-2, and that it strongly promotes the clinical effects of rhBMP-2 in bone tissue regeneration.

Animals↗