PURPOSE: This study was performed to evaluate the expressions of osteopontin (OPN) and clusterin in a transitional cell carcinoma (TCC) of the urinary bladder, and then compare their expression rates with the tumor invasiveness. MATERIALS AND METHODS: Twenty-five superficial and 25 invasive TCC were used for immunohistochemical staining. RESULTS: All 25 non-invasive TCC showed a strong positive reaction for OPN. Twenty of the invasive TCC showed a strong positive reaction for OPN, but 5 showed only a weak positive reaction. OPN expression was significantly decreased in the invasive TCC (p=0.02). Eighteen superficial TCC showed a weak positive reaction for clusterin, with 7 showing a negative reaction. Nine invasive TCC showed a strong positive reaction for clusterin, and 11 showed only a weak positive reaction. Five invasive TCC showed a negative reaction for clusterin. Clusterin expression was significantly increased in the invasive TCC (p=0.001). CONCLUSIONS: These results may suggest that OPN and clusterin could be used as markers to predict the biological behavior of a TCC.
Carcinoma, Transitional Cell* ; Clusterin* ; Osteopontin* ; Urinary Bladder*

BACKGROUND: Although basal cell carcinoma (BCC) is the most common human skin tumor, with calcification reportedly taking place in about 20% of all BCC cases, the pathogenesis of calcification in BCC has not yet been studied. OBJECTIVE: The purpose of this study was to evaluate factors related to the pathogenesis of calcification in BCC. METHODS: We performed immunohistochemical staining for beta-catenin, frizzled-2, BMP-4, osteopontin, osteocalcin, and osteonectin using frozen skin tissue from 15 cases of BCC with calcification and 11 cases of BCC without calcification. RESULTS: The expression of beta-catenin showed positive in 14 of the 15 cases in BCC with calcification, but negative in all 11 cases of BCC without calcification. The expression of frizzled-2 was observed in 14 of the 15 cases in BCC with calcification, and in 10 of the 11 cases in BCC without calcification. The difference did not reach statistical significance (p=0.236). The expression of BMP-4 was observed in all 26 samples of BCC, but the intensity of expression did not reach statistical significance between the two groups (p=0.293). Furthermore, osteopontin and osteocalcin showed no statistical significance between two the groups (p=0.567, p=0.401). The expression of osteonectin was observed in all of the BCC cases, and was stronger in BCC with calcification than in BCC without calcification (p=0.042). CONCLUSION: We suggest that the calcification in BCC might be related to the increase of beta-catenin expression and that osteonectin might also influence the process of calcification in BCC.
beta Catenin* ; Carcinoma, Basal Cell* ; Humans ; Osteocalcin* ; Osteonectin* ; Osteopontin* ; Skin

PURPOSE: This study was aimed at evaluating the histological changes of new bone and expression of osteopontin (OPN) after mandibular distraction osteogenesis. MATERIALS AND METHODS: Unilateral mandibular distraction (0.5 mm twice per day for 10 days) was performed in eight adult dogs. Two animals were sacrificed at 7, 14, 28 and 56 days after completion of distraction, respectively. The distracted bones and contralateral non-distracted control bones were harvested and processed for histological and immunohistochemical examinations. RESULTS: The new bone was arranged to tension direction after distraction osteogenesis. 7 days after distraction, numerous osteoblasts lining the immature trabecular bone and fibroblast-like cells in the fibrous intrezone were observed. 14 days after distraction, the new formed trabecular bones were thickened compared with 7 days after distraction. 28 days after distraction, the fibrous interzone was almost filled with newly calcified bone, and it was more hardened at 56 days after distraction. Increased OPN signals detected in the osteoblasts lining the trabecular bone and fibroblast-like cells in the fibrous interzone at 7 and 14 days after distraction. At 28 days after distraction, the OPN was weakly expressed in the osteoblasts, and it was not detected in all cellular components of distracted bone at 56 days later of distraction. CONCLUSIONS: After distraction osteogenesis, the distracted zone was completely calcified during the 56 days of consolidation period. In this study, the staining intensity of OPN increased in the osteoblasts and fibroblast-like cells at 7 and 14 days after distraction. The expression pattern of this protein shown here suggested that OPN play an important role in the osteogenesis during the early consolidation period.
Adult ; Animals ; Dogs ; Humans ; Osteoblasts ; Osteogenesis ; Osteogenesis, Distraction* ; Osteopontin*

Coronary Restenosis ; metabolism ; Heart Valve Diseases ; Humans ; Osteopontin ; metabolism

The purpose of this study was to evaluate the effects of DFPR on differentiation of mouse calvarial cell in vitro, to examine the possibility for periodontal regeneration. 10microgram/ml of DFPR was used as experimental concentration. osteogenic medium only was assigned as control, Experimental 1 was supplemented with 10nM dexamethasone, Experimental 2 with 10microgram/ml DFPR and Experimental 3 with 10nM dexamethasone + 10microgram/ml DFPR. cellular activity was evaluated by MTT method at 8, 12, 16 days, expression of mRNA of ALP, osteopontin, osteocalcin, collagen type-1 was detected by RT-PCR method at 4, 8, 12, 16 days of culture . extent of mineralization was observed by Von Kossa staining at 16 day of culture. The results are as follows 1)Any acceleration of differentiation was not observed at expression of differentiation marker, 2) Decrease in expression of extracelluar matrix and in bone nodule formation was observed The results suggested that DFPR have negative effect on the rate of differentiation on rat calvarial cell, decrease extracelluar matrix formation ,decrease bone nodule formation. Ongoing studies are necessary in order to determine effect of DFPR on periodontal regeneration.
Acceleration ; Animals ; Collagen ; Dexamethasone ; Methylene Chloride* ; Mice* ; Osteocalcin ; Osteopontin ; Rats ; Regeneration ; RNA, Messenger

Silica is known as a promising osteoconductive material, and polycaprolactone is a bioactive and degradable material. The purpose of this study was to monitor the differentiation of MC3T3-E1 cells cultured on the layer-built silica/poly caprolactone non-woven fabric produced by electrospinning. Non-woven fabric (silica, polycaprolactone, PSP, SPS) was made by electrospinning and they were inserted in the 48 well cell culture plate. MC3T3-E1 cells were prepared by subculture. Cells were seeded to each well 1x10(5) concentration per well. Dulbecco's modified eagle medium with 10% FBS and 1% antibiotic-antimycotic solution was used. Confocal laser scanning microscope was taken 4 hours after incubation (95% air, 5% CO2, 37degrees C). Cell proliferation was monitored by spectrophotometer on 1, 7, 14 days, and the morphology of the growing cells was observed by field emission scanning electron microscope. To monitor the differentiation of osteoblasts on the materials, MC3T3-E1 cells were incubated in 48 well culture plate after seeding with the density of 1x10(5) concentration. Then ELISA kit & EIA kit were used on to assess osteocalcin and osteopontin expression respectively. The other conditions were the same as above. MC3T3-E1 cells were proliferated well on all of the materials. There were no statistical differences among them. The osteopontin expression of silica, PSP, SPS was significantly higher than other groups on day 3 (p<0.05), but after that time, there were no statistically signigicant differences. The osteocalcin expression was significantly higher in silica and PSP than other groups on day 14. These findings show that PSP was as good as silica on the effect of osteoblast differentiation. The PSP non-woven fabric may have the possibility as bone graft materials.
Cell Culture Techniques ; Cell Proliferation ; Eagles ; Enzyme-Linked Immunosorbent Assay ; Osteoblasts ; Osteocalcin ; Osteopontin ; Silicon Dioxide ; Transplants

Osteopontin (OPN) is a secreted glycoprotein that is expressed in various tissues, including brain, and mediates a wide range of cellular activities. In a previous study, the authors observed the robust neuroprotective effects of recombinant OPN and of RGD and SLAYGLR-containing OPN-peptide icosamer (OPNpt20) in an animal model of transient focal ischemia, and demonstrated anti-inflammatory and pro-angiogenic effects of OPNpt20 in the postischemic brain. In the present study, we investigated the effects of OPNpt20 on the motility and phagocytic activity of BV2 cells (a microglia cell line). F-actin polymerization and cell motility were significantly enhanced in OPNpt20-treated BV2 cells, and numbers of filopodia-like processes increased and lamellipodia-like structures enlarged and thickened. In addition, treatment of cells with either of three mutant OPN icosamers containing mutation within RGD, SLAY, or RGDSLAY showed that the RGD and SLAY motifs of OPNpt20 play critical roles in the enhancement of cell motility, and the interaction between exogenous OPNpt20 and endogenous αv and α4 integrin and the activations of FAK, Erk, and Akt signaling pathways were found to be involved in the OPNpt20-mediated induction of cell motility. Furthermore, phagocytic activity of microglia was also significantly enhanced by OPNpt20 in a RGD and SLAY dependent manner. These results indicate OPNpt20 containing RGD and SLAY motifs triggers microglial motility and phagocytic activity and OPNpt20-integrin mediated signaling plays a critical role in these activities.
Actins ; Brain ; Cell Movement ; Glycoproteins ; Ischemia ; Microglia* ; Models, Animal ; Neuroprotective Agents ; Osteopontin* ; Phagocytosis ; Polymerization ; Polymers

OBJECTIVE: To evaluate the effects of both exogenous and endogenous osteopontin on normal and malignant ovarian epithelial cell growth, and on paclitaxel chemo-resistance. METHODS: The ovarian cancer cell line OV429, which showed low level of endogenous osteopontin and paclitaxel sensitive cell line OV420, which showed high level of endogenous osteopontin, and a normal ovarian epithelial (HOSE: Human ovarian surface epithelial) cells were treated with purified osteopontin. Furthermore, OV420 was treated with osteopontin siRNA alone or in combination with paclitaxel. Proliferation rates and cell cycle progression of treated cells were determined by the tetrazolium colorimetric (XTT) assay and FACS analysis, respectively. RESULTS: Exogenous osteopontin increased the proliferation rate of OV429 and OV420 but had negligible effect on normal HOSE. Ovarian cancer cell lines treated with siRNA showed significantly reduced the growth rates (P<0.05), and they were arrested in G2/M phase of the cell cycle. Furthermore, OV420 treated with paclitaxel in the presence of osteopontin siRNA showed significantly decreased the survival rate. CONCLUSION: Osteopontin promote cell growth in malignant but not in normal ovarian epithelial cells, and may confer paclitaxel-resistance by adhesion to each cell and minimized the cell surface which exposure to chemo-agents.
Cell Cycle ; Cell Line ; Epithelial Cells* ; Humans ; Osteopontin* ; Ovarian Neoplasms ; Paclitaxel ; RNA, Small Interfering ; Survival Rate

PURPOSE: To characterize the effects of 2-deoxy-D-glucose (2-DG) and quercetin (QCT) on gene expression of osteonectin (ON) and osteopontin (OP) in irradiated MC3T3-E1 cells. MATERIALS AND METHODS: When MC3T3-E1 osteoblastic cells had reached 70-80% confluence, cultures were transferred to a differentiating medium supplemented with 5 mM 2-DG or 10 micrometer QCT and then irradiated with 2, 4, 6, and 8 Gy. At various times after irradiation, the cells were analyzed for the expression of bone mineralization genes such as ON and OP. RESULTS: The mRNA expression of both ON and OP was increased according to the culture time in the differentiation medium, and the increase of the genes peaked at 14 days after the differentiation induction. In the case of OP, the increase of mRNA expression was maintained to 28 days after the differentiation, while the mRNA level of ON was reduced to the basal level at the same time. Irradiation adding 2-DG showed a significant peak value in the expression pattern of ON at 4 Gy 7 days after irradiation. Irradiation adding QCT increased the mRNA expression of ON and OP in a dose-dependant manner, but irradiation adding 2-DG did not show any differences between the control and experiments 14 days after irradiation. Irradiation adding QCT increased significantly the expression patterns of ON 21 days after irradiation. CONCLUSION: The results showed that QCT acted as a radiosensitizer in the gene expression of ON and OP during differentiation of the late stage of irradiated MC3T3-E1 osteoblastic cells in vitro.
Calcification, Physiologic ; Deoxyglucose ; Gene Expression ; Osteoblasts ; Osteonectin ; Osteopontin ; Quercetin ; RNA, Messenger

PURPOSE: In this paper we tried to evaluate the most appropriate surface for rhBMP-2 coating among 4 rough titanium surfaces. MATERIALS AND METHODS: We used machined surface as a control group and anodized, RBM and SLA surfaces as test groups. We coated rhBMP-2 on the 4 surfaces and with uncoated surfaces for each case, we cultured human mesenchymal stem cells on all 8 surfaces. 24 hours after we measured the stem cell'attachment with SEM, and on 3rd, 7th, and 14th days, we checked the cell proliferation and differentiation by using MTT and ALP activity assay. And on the 7th day after the culture, we performed RT-PCR assay to determine whether the expression levels of Type I collagen, osteocalcin, osteopontin were changed. RESULTS: We observed with SEM that 4 rhBMP-2 coated surfaces exhibited wider and tighter cell attachment and more cell process spreading than uncoated surfaces. The anodized rhBMP-2 surface caused robustest effects. In MTT assay we could not find any meaningful difference. In ALP assay there was a significant increase (P<.05) in the ALP activity of anodized rhBMP-2 coated surface compared with that of the control (3rd and 14th days) and with that of the RBM rhBMP-2 coated surface (14th day). In RT-PCR assay there was increased expressions in the anodized rhBMP-2 coated surface for osteocalcin, and osteopontin. CONCLUSION: We found that the anodized rhBMP-2 coated surface were most prominent stem cell attachment and differentiation in compared to control and Machined rhBMP-2 coated, RBM rhBMP-2 coated surface.
Cell Proliferation ; Collagen Type I ; Humans ; Mesenchymal Stromal Cells ; Osteocalcin ; Osteopontin ; Stem Cells ; Titanium