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Biomedical subjects

C Hamanishi

Publications and source records attributed to C Hamanishi.

At least 19 recordsLinked to original sources
PubMed2003

Cyclic tensile stretch loaded on bovine chondrocytes causes depolymerization of hyaluronan: involvement of reactive oxygen species.

OBJECTIVE: We have previously demonstrated that reactive oxygen species (ROS) are involved in cartilage degradation. Decreased size of hyaluronan (HA), the major macromolecule in synovial fluid, to which it imparts viscosity, is reported in patients with arthritis. The purpose of this study was to determine the alteration in the molecular weight range of HA as a result of mechanical deformation loaded on the chondrocytes, as well as the involvement of ROS in this action. METHODS: ROS were generated via the oxidation of hypoxanthine by xanthine oxidase. Cyclic tensile stretch was loaded using a vacuum-operated instrument. Levels of HA were measured using a sandwich enzyme-binding assay. Superoxide dismutase (SOD) activity and ROS were measured using water-soluble tetrazolium and a chemiluminescent probe, respectively. RESULTS: ROS depolymerized HA molecules. Cyclic tensile stretch depolymerized HA and induced ROS. SOD inhibited not only ROS induction but also HA depolymerization caused by the mechanical stress. CONCLUSION: ROS play an important role in mechanical stress-induced HA depolymerization.

Animals↗
PubMed2002

Hydrogen peroxide induces apoptosis of osteocytes: involvement of calcium ion and caspase activity.

We hypothesized that reactive oxygen species play an important role in avascular/ischemic osteonecrosis. When isolated chick osteocytes were cultured with hydrogen peroxide, annexin V binding, which is the earliest marker of apoptosis, increased in a dose-dependent fashion. Hydrogen peroxide also induced the activation of caspase-3 and increase in cytosolic Ca2+. Treatment with BAPTA/AM (cheletor of cytosolic Ca2+) and Ac-DEVD-cho (caspase inhibitor) attenuated hydrogen peroxide-induced apoptosis. These data demonstrated the signal transduction pathways that participate in this hydrogen peroxide-induced cell damage.

Animals↗
PubMed2001

Cyclic tensile stretch inhibition of nitric oxide release from osteoblast-like cells is both G protein and actin-dependent.

Recent reports indicate the alteration of nitric oxide (NO) synthesis with mechanical stress loaded on the osteoblast and NO is considered to have a significant role in mechanotransduction. We found the involvement of guanine-nucleotide-binding regulatory proteins (G proteins), especially Gi, in stress-inhibited NO release of osteoblast-like cells (JOR:17;593-597, 1999). To determine further the mechanism involved in this process, we measured c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) activity under cyclic tensile stretch loaded on osteoblast-like cells. Cyclic stretch significantly enhanced JNK/SAPK activity and pertussis toxin clearly reversed stress-enhanced JNK/SAPK activity. Cytochalasin D, actin microfilament disrupting reagent, also abolished the stress activation of JNK/SAPK. We propose a model for signaling events induced by cyclic tensile stretch, namely a transmembrane mechanosensor which couples Gi-protein, actin cytoskeleton and finally activates JNK/SAPK activity of osteoblasts.

Actins↗
PubMed2001

Interleukin-4 reversed the Interleukin-1-inhibited proteoglycan synthesis through the inhibition of NO release: a possible involvement of intracellular calcium ion.

Interleukin-1 (IL-1) causes cartilage degradation through nitric oxide (NO) synthesis. Although Interleukin-4 (IL-4) antagonizes the IL-1-mediated cartilage degradation, the precise mechanisms are not clear. We examined the effect of IL-4 on NO synthesis in parallel with intracellular Ca levels ([Ca(2+)]i) and proteoglycan (PG) synthesis. IL-4-inhibited IL-1-enhanced NO release in a dose-dependent manner. IL-1-enhanced [Ca(2+)]i in the chondrocytes, and IL-4 attenuated this increase. IL-4 reversed IL-1-inhibited PG synthesis. Accordingly, IL-4 reversed the IL-1-inhibited PG synthesis through the inhibition of NO release. An increase in [Ca(2+)]i with IL-1 is possibly involved in this action.

Journal Article↗
PubMed2000

Hyaluronic acid inhibits the expression of u-PA, PAI-1, and u-PAR in human synovial fibroblasts of osteoarthritis and rheumatoid arthritis.

OBJECTIVE: Intraarticular administration of hyaluronic acid (HA) has been widely used for the treatment of osteoarthritis (OA). Fibrinolysis is closely related to the pericellular proteolysis involved in inflammation. However, the role of HA in the regulation of fibrinolytic factors is not yet known. We investigated the effect of HA on the pericellular fibrinolytic system of human synovial fibroblasts derived from OA and rheumatoid arthritis (RA). METHODS: Human synovial fibroblasts obtained from OA and RA were cultured in the presence and absence of HA. The antigen of urokinase-type plasminogen activator (u-PA) and plasminogen activator inhibitor-1 (PAI-1) were measured by ELISA, and u-PA activity was evaluated by electrophoretic enzymography. The binding assay of u-PA and the immunohistochemical analysis of u-PA were employed to detect u-PA receptor (u-PAR). RESULTS: HA suppressed the secretion of both u-PA and PAI-1 antigens from the synovial fibroblasts of OA to their conditioned medium. Suppression of u-PA activity in OA synovial fibroblasts was more marked than in those of RA. The u-PA binding assay of OA and RA synovial fibroblasts revealed a single class of binding site: dissociation constant (Kd) 23.7 nM, maximal number of binding sites (Bmax) 3.11x10(4) binding sites/cell; Kd 16.5 nM, Bmax of 9.88x10(4) binding sites/cell, respectively. HA decreased Bmax in fibroblasts of both OA and RA. Immunohistochemical analysis showed that u-PAR was constitutively expressed in both synovial fibroblasts, but if these cells were treated with HA, the decrease of the staining of u-PAR was more pronounced in the cells of RA than in OA. CONCLUSION: Pericellular fibrinolytic activity mediated by the u-PA/u-PAR system and PAI-1 was attenuated by HA in synovial fibroblasts derived from OA and RA. Thus, HA may be a useful agent to inhibit the inflammation of arthritis.

Adjuvants, Immunologic↗
PubMed1999

Healing of segmental bone defects in rats induced by a beta-TCP-MCPM cement combined with rhBMP-2.

A beta-tricalcium phosphate-monocalcium phosphate monohydrate (beta-TCP-MCPM) cement was evaluated as an effective carrier of recombinant human bone morphogenetic protein-2 (rhBMP-2) in rat femoral critical-size defects. Hard cement cylinders (4 x 5 mm) impregnated with two different doses of rhBMP-2 (1.26 or 6.28 microg) were implanted into each defect, and the results were compared with those in rats that had implantations of cylinders only. Implantation of the 6.28 microg dose of rhBMP-2 caused a large bone shell to form around the defect, resulting in osseous union in all cases within 3 weeks. Except for beta-TCP granules, the cement was resorbed and replaced by bone tissue at 6 weeks. A torsion test at 9 weeks showed that the failure torque and bone stiffness had recovered 99% and 141%, respectively, compared with the intact contralateral femur. The defects that received 1.26 microg of rhBMP-2 resulted in 40% union and 41% of the failure torque at 9 weeks. However, no instances of union were observed in the defects implanted with cylinders only. In conclusion, the beta-TCP-MCPM cement was shown to be effective as a rhBMP-2 carrier. Combined with rhBMP-2, this cement was rapidly resorbed and completely healed the defects.

Animals↗
PubMed1999

Pertussis toxin-sensitive G proteins as mediators of stretch-induced decrease in nitric-oxide release of osteoblast-like cells.

Mechanical loading plays an important role in regulating bone remodeling, and nitric oxide may be one regulator of this process. To determine how mechanical stress modulates osteoblast function, we loaded cyclic tensile stretch on osteoblast-like cells and measured levels of nitric oxide in the medium. High frequency of stretch at any magnitude inhibited release of nitric oxide; however, low frequency of stretch enhanced its release from the static control. To examine the involvement of G protein (guanine nucleotide-binding regulatory protein) in stress-inhibited release of nitric oxide, we added pertussis toxin, a specific inhibitor of the Gi class, and found that it completely reversed the stress-inhibited release. These data support the idea that pertussis toxin-sensitive G protein is activated in the presence of cyclic tensile stretch.

Bone Remodeling↗
PubMed1999

Effect of hydrogen peroxide on the metabolism of articular chondrocytes.

OBJECTIVE: To examine the effect of hydrogen peroxide on chondrocyte metabolism. MATERIALS AND METHODS: Bovine articular chondrocytes were used. Proteoglycan (PG) synthesis was measured with [35S] sulfate incorporation. For detection of apoptosis, the TdT-mediated dUTP-biotin nick end labeling (TUNEL) and annexin V assay were used. Extracellular-regulated protein kinase (ERK) activity was measured using a mitogen-activated protein kinase assay system. RESULTS: Addition of hydrogen peroxide resulted in the inhibition of PG synthesis, apoptosis, and enhanced ERK activity. CONCLUSION: Hydrogen peroxide plays an important role in regulating the metabolism of chondrocytes.

Animals↗
PubMed1998

Auto-destruction of the articular cartilage and free radical mediators.

The effect of static compression on the release of superoxide (SO) and nitric oxide (NO) from cartilage obtained from rabbit knee joints with tissue defects was studied. The rabbits were divided into two groups: (1) those that had 5 mm diameter full chondral defects (defect group) and (2) those in which chondral defects were filled with autogenous perichondrial grafts (grafted group). Histologically, cartilage was regenerated in the grafted group 3 weeks after the operation, although only a fibrous tissue filled the defects in the defect group even 16 weeks after the operation. A static pressure of 5 kg for 10 minutes applied to the cartilage chips obtained from the area surrounding the defects released significant amounts of SO and NO into the medium. Maximum increases were observed in the defect group 3 weeks after the operation.

Animals↗
PubMed1998

Factors related to degradation of articular cartilage in osteoarthritis: a review.

OBJECTIVES: Osteoarthritis (OA) is a common joint deterioration initiated by multiple factors. To better understand related factors in the development of this disease, we focused on the mechanical stress loaded on articular cartilage. MATERIALS AND METHODS: The anterior cruciate ligaments of rabbit knee joints were transected, and expression of protein kinase C (PKC) examined immunohistochemically. The PKC activator 12-o-tetradecanoyl-phorbol-13-acetate (TPA) was then administered intraarticularly. To determine the involvement of gas mediators, a cartilage defect was made on the medical femoral condyle of rabbit knee joints. Hydrostatic pressure was loaded on the cartilage taken from the surrounding defects, and levels of superoxide anion and nitric oxide (NO) were measured. Bovine chondrocytes were subjected to cyclic mechanical stretch using a Flexercell Strain Instrument. Proteoglycan synthesis and PKC activity were measured. Expression of matrix metalloproteinase (MMP)-3 and tissue inhibitor of metalloproteinase (TIMP)-1 in articular cartilages obtained from OA patients were examined using Northern blots. RESULTS: Chondrocytes from experimentally induced OA were stained positively with anti-alpha-PKC antibody. Intraarticular administration of TPA prevented the development of OA changes. Cyclic tensile stretch loaded on chondrocytes decreased proteoglycan synthesis and PKC activity. Thus, PKC is involved in the stress-mediated degradation of articular cartilage. Cartilage defects led to degradation of surrounding cartilage and to enhanced superoxide anion and NO synthesis. We also noted increased and decreased expressions of MMP-3 and TIMP-1 mRNA in human OA cartilage, respectively. CONCLUSION: PKC, gas mediators (superoxide anion, NO), and proteinases are all involved in OA.

Animals↗
PubMed1996

Self-setting, bioactive, and biodegradable TTCP-DCPD apatite cement.

Tetracalcium phosphate-dicalcium phosphate dihydrate self-setting apatite cement mixed with low-crystallized seed hydroxyapatite is similar to the host bone in degree of crystallinity. The bonding strength of this cement with hydroxyapatite-coated titanium rods was twice that with noncoated titanium and four times that with stainless steel. Histologically, TTCP-DCPD apatite cement incorporated into the tibia of rabbits were degraded, absorbed, and replaced by the normal body trabeculae rapidly. The mechanical strength of the cement disk intercalated into a gap made in the rabbit tibiae increased to 73% of that of the normal tibia at 10 weeks concurrently with the decrease in bone mineral density of the disk toward that of the normal tibia, in which bony replacement of the disk was observed also histologically. These bioactive and biodegradative characters of this cement are due to the similarity of its degree of crystallinity to that of the host bone, and could expand its clinical applications.

Animals↗
PubMed1996

A self-setting TTCP-DCPD apatite cement for release of vancomycin.

Vancomycin (VCM), a methiciline-cefem resistant Staphylococcus aureus (MRSA)-specific antibiotic, was incorporated in a self-setting tetracalcium phosphate (TTCP)-dicalcium phosphate dihydrate (DCPD) apatite cement that hardened isothermally into a hydroxyapatite (HAP) phase with crystallinity similar to that of host bone. Effective release of VCM into PBS lasted for 2 weeks from cements containing 1% VCM and for longer than 9 weeks from cements containing 5% VCM. The rate of release of VCM differed between cements with different crystallinities as well as between the two dissolution media, PBS and simulated body fluid. Mean concentration of VCM in the bone marrow tissue released from cements containing 5% VCM was 20 times the minimum inhibitory concentration 3 weeks after implantation in bone. Direct contact with new bone was observed with the cements containing 1% VCM. Slow delivery of VCM from a self-setting TTCP-DCPD apatite cement with low crystallinity could be used to treat MRSA osteomyelitis.

Animals↗
PubMed1996

Alpha and epsilon isozymes of protein kinase C in the chondrocytes in normal and early osteoarthritic articular cartilage.

Protein kinase C (PKC) isozymes (alpha, beta I, beta II, gamma and, epsilon-PKC) were examined immunocytochemically in control and mechanically induced osteoarthritic knee joints in rats. beta I, beta II, and gamma PKC-positive cells were not observed in sham-operated or osteoarthritic knee joints. alpha-PKC, which was observed only in the cells in the subchondral bony layer in the controls, appeared in the chondrocytes in the superficial and columnar layers of the osteoarthritic knees. epsilon-PKC was observed in the control chondrocytes in the superficial portion of the columnar layers. In early osteoarthritic joints, however, epsilon-PKC-positive chondrocytes disappeared from the superficial portion of columnar layer and increased in number in the middle columnar layers. The appearance and changes in distribution of these PKC-positive chondrocytes, either under normal or osteoarthritic conditions, have not been reported previously. The role of the redistribution of PKC isozymes is still unclear as to whether they are involved in initiating destructive processes, reflect attempted cell repair mechanisms, or are simply a consequence of the cellular changes.

Animals↗
PubMed1996

Protein kinase C activator inhibits progression of osteoarthritis induced in rabbit knee joints.

To study the role of protein kinase C (PKC) in cartilage tissue in osteoarthritis, experimental osteoarthritis was induced in the knee joints of rabbits by resection of the anterior cruciate ligament (ACL). At 4 weeks after the operation, osteoarthritic changes varying from surface irregularities and cleft formation to loss of the tangential layer were observed, and cloning or hypocellularity of the chondrocytes was observed mainly in the transitional and radial layers. The PKC activator 12-O-tetradecanoyl-phorbol-13-acetate (TPA) or non-PKC-activating phorbol ester 4 alpha-phorbol-12,13-didecanate (PDD) was administered intraarticularly once a week from the day of the operation for 3 weeks. Histologic evaluation with a rating scale was carried out. In the TPA-administered group, cartilage structures were preserved almost completely, and score of the cartilage lesion was significantly less than that in animals administered PDD or in nonadministered controls. A chondroprotective role of PKC under mechanical stress was suggested.

Animals↗
PubMed1996

Bilateral multilevel laminectomy with or without posterolateral fusion for cervical spondylotic myelopathy: relationship to type of onset and time until operation.

The authors retrospectively evaluated the relationship of several preoperative factors in 69 patients who had myelopathy due to multilevel cervical spondylosis without ossification of the posterior longitudinal ligament treated with Kirita's bilateral wide laminectomy. In 34 patients with focal instability or malalignments, posterolateral fusion was also combined. The clinical results at an average follow-up period of 3.5 years (range 1-10 years) after operation in the groups that had and had not undergone fusion were equally satisfactory, and preoperative focal instability was believed to be the sole useful indication for adding posterolateral fusion. The patients were classified in three groups according to the acuteness of the onset. The type of onset and time until operation were found to be the factors most strongly related to prognosis, and clinical outcome was correlated with the duration after onset when plotted as days in the acute, months in the subacute, and years in the insidious onset groups. Wide laminectomy with or without posterolateral fusion is a simple operation that is recommended, provided that it is performed early enough according to the type of onset.

Aged↗
PubMed1995

Ossification of the posterior longitudinal ligament. Autosomal recessive trait.

STUDY DESIGN: This study analyzed the mode of inheritance of ossification of the posterior longitudinal ligament (OPLL) from the pedigree of a family. OBJECTIVES: The results were correlated to provide a new mode of inheritance of OPLL. SUMMARY OF BACKGROUND DATA: Although a nation-wide multicenter survey of OPLL has been carried out in 347 subjects and 1030 relatives in Japan since 1981, no parental consanguinity has been reported. METHODS: In the family, three siblings and one other member of a family underwent operations for OPLL in our department, and the clinical information regarding other family members was obtained from interviews with these four patients or relatives. RESULTS: The parents of three affected siblings and another unaffected sister were first cousins, and the father was suspected to be affected as well. CONCLUSION: Transmission of OPLL as an autosomal recessive trait in this family, which has not been reported, is suspected; although, the possibility that it is a dominant trait can not be excluded.

Aged↗
PubMed1995

Bone mineral density changes in distracted callus stimulated by pulsed direct electrical current.

To evaluate the effect of application of pulsed direct current electrical stimulation to callus tissue, a 1-cm bone-lengthening model using an external lengthener was applied to rabbit tibia. Twenty-microampere pulsed direct current was applied 12 hours daily from the day of osteotomy until 40 days after the completion of lengthening. The area, bone mineral content, and bone mineral density of the distracted callus and of the proximal and distal segments of the tibia were evaluated using dual-energy xray absorptiometry. The absolute and relative values of bone mineral density of the electrically stimulated callus were significantly increased as compared with those in the control group. Pulsed direct current electrical stimulation may be indicated in bone lengthening to stimulate the poorly mineralized callus, and may shorten the overall time course of leg lengthening.

Absorptiometry, Photon↗