Chien-Chung Kuo

Thrombin enhanced migration and MMPs expression of human chondrosarcoma cells involves PAR receptor signaling pathway

Thrombin is a multifunctional protease that can activate hemostasis and coagulation through the cleavage of fibrinogen to form fibrin clots. Thrombin also plays a crucial role in migration and metastasis of human cancer cells. However, the effect of thrombin on migration activity in human chondrosarcoma cells is mostly unknown. Here, we found that thrombin increased the migration and expression of matrix metalloproteinase (MMP)‐2 and MMP‐13 in human chondrosarcoma cells (JJ012 and SW1353 cells). By using pharmacological inhibitors or activators or genetic inhibition by the protease‐activated receptor (PAR), we found that the PAR1 and PAR4 receptor but not PAR3 receptor are involved in thrombin‐mediated cell migration and MMPs expression. Thrombin‐mediated migration and MMPs up‐regulation was attenuated by phospholipase C (PLC), protein kinase C, and c‐Src inhibitor. Activations of PLCβ, PKCα, c‐Src, and NF‐κB pathways after thrombin treatment was demonstrated, and thrombin‐induced MMPs expression and migration activity was inhibited by the specific inhibitors and mutants of PLC, PKC, c‐Src, and NF‐κB cascades. Taken together, our results indicated that thrombin enhances the migration of chondrosarcoma cells by increasing MMP‐2 and MMP‐13 expression through the PAR/PLC/PKCα/c‐Src/NF‐κB signal transduction pathway. J. Cell. Physiol. 223:737–745, 2010.

CCN3 increases cell motility and MMP-13 expression in human chondrosarcoma through integrin-dependent pathway.

Chondrosarcoma is a type of highly malignant tumor with a potent capacity to invade locally and cause distant metastasis. Chondrosarcoma shows a predilection for metastasis to the lungs. CCN3, also called nephroblastoma overexpressed gene (NOV), regulates proliferation and differentiation of cancer cells. However, the effect of CCN3 on migration activity in human chondrosarcoma cells is mostly unknown. Here, we found that CCN3 increased the migration and expression of matrix metalloproteinase (MMP)‐13 in human chondrosarcoma cells (JJ012 cells). αvβ3 or αvβ5 monoclonal antibody (mAb), phosphatidylinositol 3‐kinase (PI3K) inhibitors (Ly294002 and wortmannin) and Akt inhibitor inhibited the CCN3‐induced increase of the migration and MMP‐13 upregulation of chondrosarcoma cells. CCN3 stimulation increased the phosphorylation of focal adhesion kinase (FAK), PI3K, and Akt. In addition, NF‐κB inhibitors also suppressed the cell migration and MMP‐13 expression enhanced by CCN3. Moreover, CCN3 increased NF‐κB luciferase activity and binding of p65 to the NF‐κB element on the MMP‐13 promoter. Taken together, our results indicate that CCN3 enhances the migration of chondrosarcoma cells by increasing MMP‐13 expression through the αvβ3/αvβ5 integrin receptor, FAK, PI3K, Akt, p65, and NF‐κB signal transduction pathway. J. Cell. Physiol. 226: 3181–3189, 2011.

CCN3 increases BMP-4 expression and bone mineralization in osteoblasts

The nephroblastoma overexpressed (NOV) gene, also called CCN3, regulates differentiation of skeletal mesenchymal cells. Bone morphogenetic proteins (BMPs) play important roles in osteoblast differentiation and bone formation, but the effects of CCN3 on BMP expression and bone formation in cultured osteoblasts are largely unknown. Here we found that CCN3 increased BMP‐4 expression and bone nodule formation in cultured osteoblast. Monoclonal antibodies for α5β1 and αvβ5 integrins, and inhibitors of integrin‐linked kinase (ILK), p38, and JNK, all inhibited CCN3‐induced bone nodule formation and BMP‐4 up‐regulation of osteoblasts. CCN3 stimulation increased the kinase activity of ILK and phosphorylation of p38 and JNK. Inhibitors of activator protein‐1 (AP‐1) also suppressed bone nodule formation and BMP‐4 expression enhanced by CCN3. Moreover, CCN3‐induced c‐Jun translocation into the nucleus, and the binding of c‐Jun to the AP‐1 element on the BMP‐4 promoter were both inhibited by specific inhibitors of the ILK, p38, and JNK cascades. Taken together, our results provide evidence that CCN3 enhances BMP‐4 expression and bone nodule formation in osteoblasts, and that the integrin receptor, ILK, p38, JNK, and AP‐1 signaling pathways may be involved. J. Cell. Physiol. 227: 2531–2541, 2012.

Involvement of integrin up‐regulation in RANKL/RANK pathway of chondrosarcomas migration

Invasion of tumor cells is the primary cause of therapeutic failure in malignant chondrosarcomas treatment. Receptor activator of nuclear factor‐κB ligand (RANKL) and its receptor, RANK, play a key roles in osteoclastogenesis and tumor metastasis. We found that the RANKL and RANK expression in human chondrosarcoma tissues was higher than that in normal cartilage. We also found that RANKL directed the migration and increased cell surface expression of β1 integrin in human chondrosarcoma cells (JJ012 cells). Pretreatment of JJ012 cells with MAPK kinase (MEK) inhibitors, PD98059 or U0126, inhibited the RANKL‐induced migration and integrin expression. Stimulation of cells with RANKL increased the phosphorylation of MEK and extracellular signal‐regulating kinase (ERK). In addition, NF‐κB inhibitor (PDTC) or IκB protease inhibitor (TPCK) also inhibited RANKL‐induced cells migration and integrin up‐regulation. Taken together, these results suggest that the RANKL acts through MEK/ERK, which in turn activates IKKα/β and NF‐κB, resulting in the activation of β1 integrin and contributing to the migration of human chondrosarcoma cells. J. Cell. Biochem. 111: 138–147, 2010.

Three-dimensional computer graphics-based ankle morphometry with computerized tomography for total ankle replacement design and positioning.

Morphometry of the bones of the ankle joint is important for the design of joint replacements and their surgical implantations. However, very little three‐dimensional (3D) data are available and not a single study has addressed the Chinese population. Fifty‐eight fresh frozen Chinese cadaveric ankle specimens, 26 females, and 32 males, were CT‐scanned in the neutral position and their 3D computer graphics‐based models were reconstructed. The 3D morphology of the distal tibia/fibula segment and the full talus was analyzed by measuring 31 parameters, defining the relevant dimensions, areas, and volumes from the models. The measurements were compared statistically between sexes and with previously reported data from Caucasian subjects. The results showed that, within a general similarity of ankle morphology between the current Chinese and previous Caucasian subjects groups, there were significant differences in 9 out of the 31 parameters analyzed. From a quantitative comparison with available prostheses designed for the Caucasian population, few of these designs have both tibial and talar components suitable in dimension for the Chinese population. The current data will be helpful for the sizing, design, and surgical positioning of ankle replacements and for surgical instruments, especially for the Chinese population. Clin. Anat. 659–668, 2014.

Effects of positioning on radiographic measurements of ankle morphology: a computerized tomography-based simulation study.

BackgroundMeasurements of the morphology of the ankle joint, performed mostly for surgical planning of total ankle arthroplasty and for collecting data for total ankle prosthesis design, are often made on planar radiographs, and therefore can be very sensitive to the positioning of the joint during imaging. The current study aimed to compare ankle morphological measurements using CT-generated 2D images with gold standard values obtained from 3D CT data; to determine the sensitivity of the 2D measurements to mal-positioning of the ankle during imaging; and to quantify the repeatability of the 2D measurements under simulated positioning conditions involving random errors.MethodFifty-eight cadaveric ankles fixed in the neutral joint position (standard pose) were CT scanned, and the data were used to simulate lateral and frontal radiographs under various positioning conditions using digitally reconstructed radiographs (DRR).Results and discussionIn the standard pose for imaging, most ankle morphometric parameters measured using 2D images were highly correlated (R > 0.8) to the gold standard values defined by the 3D CT data. For measurements made on the lateral views, the only parameters sensitive to rotational pose errors were longitudinal distances between the most anterior and the most posterior points of the tibial mortise and the tibial profile, which have important implications for determining the optimal cutting level of the bone during arthroplasty. Measurements of the trochlea tali width on the frontal views underestimated the standard values by up to 31.2%, with only a moderate reliability, suggesting that pre-surgical evaluations based on the trochlea tali width should be made with caution in order to avoid inappropriate selection of prosthesis sizes.ConclusionsWhile highly correlated with 3D morphological measurements, some 2D measurements were affected by the bone poses in space during imaging, which may affect surgical decision-making in total ankle arthroplasty, including the amount of bone resection and the selection of the implant sizes. The linear regression equations for the relationship between 2D and 3D measurements will be helpful for correcting the errors in 2D morphometric measurements for clinical applications.

Inter-joint sharing of total support moments in the lower extremities during gait in narrow-heeled shoes of different heights

The study aimed to investigate the influence of the base and height of shoe heels on the total support moment (Ms) and individual joint contributions during gait. Fifteen healthy females walked barefoot and with narrow-heeled shoes (heel heights: 3.9, 6.3 and 7.3 cm) while kinematic and kinetic data were measured. Compared with the barefoot condition, the subjects maintained unaltered Ms in the sagittal plane in shod conditions. This was achieved by increasing the knee extensor moment to compensate for the diminished ankle plantarflexor moments in medium and high heel conditions. In the frontal plane, subjects in shod conditions had to sustain an increased Ms for balance control during late single-leg stance with increased knee abductor and ankle pronator moments as a result of the reduced base of the heels. The results will be helpful for future shoe designs to reduce fall risks and prevent relevant musculoskeletal problems Practitioner Summary: Knowledge of the influence of narrow-heeled shoes on lower limb support moments helps in shoe design to address fall risks. Gait analysis showed that females in narrow-heeled shoes maintained unaltered sagittal total support moments but sustained an increased demand in the frontal plane during late single-leg stance.

Bilateral Locked Posterior Fracture-dislocation of the Shoulders Due to Electric Shock

Bilateral locked posterior fracture-dislocation of the shoulders is very uncommon. The majority of cases are caused by seizure. We report a patient with bilateral locked posterior fracture-dislocation due to electric shock; he underwent bilateral hemiarthroplasty surgery two weeks after being electrocuted. The patient was able to return to work and was satisfied with his shoulder mobility on follow-up examination 6 years after surgery.

Gait changes in individuals with bilateral hallux valgus reduce first metatarsophalangeal loading but increase knee abductor moments.

Hallux valgus (HV), one of the most common foot pathologies in modern society, affects not only the foot itself, but also the other lower limb joints. The purpose of the study was to investigate the kinematic and kinetic changes in the lower limb joints in patients with bilateral HV during level walking. Twelve female patients with bilateral HV and 12 healthy female controls walked while three-dimensional kinematic and kinetic data were measured. Patients with HV were found to shift their center of pressure (COP) laterally away from the 1st metatarsophalangeal joint (MPJ), which helped unload the joint during late stance. The lateral shift of the COP in these patients was associated with the reduced toe-out angles of the foot as a result of increased internal rotation of the hip. However, this strategy increased the abductor moments at the knee, an index closely correlated with the medial load at the knee and a predictor of the onset and progression of medial OA. Early treatment of HV may be helpful not only for reducing foot pain and deformity, but also for preventing the potentially harmful loading at the knee, especially in those at risk of medial knee OA.

Effects of bilateral medial knee osteoarthritis on intra- and inter-limb contributions to body support during gait.

Patients with knee OA show altered gait patterns, affecting their quality of living. The current study aimed to quantify the effects of bilateral knee OA on the intra-limb and inter-limb sharing of the support of the body during gait. Fifteen patients with mild, 15 with severe bilateral knee OA, and 15 healthy controls walked along a walkway while the kinematic and kinetic data were measured. Compared with the controls, the patients significantly reduced their knee extensor moments and the corresponding contributions to the total support moment in the sagittal plane (p<0.05). For compensation, the mild OA group significantly increased the hip extensor moments (p<0.05) to maintain close-to-normal support and a more symmetrical inter-limb load-sharing during double-limb support. The severe OA group involved compensatory actions of both the ankle and hip, but did not succeed in maintaining a normal sagittal total support moment during late stance, nor a symmetrical inter-limb load-sharing during double-limb support. In the frontal plane, the knee abductor moments and the corresponding contributions to the total support moment were not affected by the changes in the other joints, regardless of the severity of the disease. The observed compensatory changes suggest that strengthening of weak hip muscles is essential for body support during gait in patients with knee OA, but that training of weak ankle muscles may also be needed for patients with severe knee OA.