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

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

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*

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

PURPOSE: Zinc, a biomineral present within and outside cells, manages various cellular mechanisms. In this study, we examined whether zinc was involved in vascular smooth muscle cell (VSMC) calcification via regulation of calcification inhibitor protein, osteopontin (OPN). METHODS: Rat aorta cell line (A7r5 cells) and primary vascular smooth muscle cells (pVSMCs) from rat aorta were cultured with phosphate (1-5 mM) and zinc (0-15 microM) as appropriate, along with osteoblasts (MC3T3-E1) as control. The cells were then stained for Ca and P deposition for calcification examination as well as osteopontin expression as calcification inhibitor protein was measured. RESULTS: Both Ca and phosphate deposition increased as the addition of phosphate increased. In the same manner, the expression of osteopontin was upregulated as the addition of phosphate increased in both cell types. When zinc was added, Ca and P deposition decreased in VSMCs, while it increased in osteoblasts. CONCLUSION: The results imply that zinc may prevent VSMC calcification by stimulating calcification inhibitor protein OPN synthesis in VSMCs.
Animals ; Aorta ; Cell Line ; Muscle, Smooth, Vascular* ; Osteoblasts ; Osteopontin ; Rats ; Zinc*

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

Osteopontin(OPN) is a secreted phosphoprotein that is expressed constitutively in the descending thin limb(DTL) and papillary surface epithelium (PSE). Although the function of OPN is not known with certainty, it has been suggested that OPN may play a role in the regulation of calcium-mediated or calcium-dependent processes. The aim of this study was to compare the effects of 1,25-dihydroxyvitamin D3(VitD), parathyroid hormone(PTH) and calcitonin, the hormones involved in the regulation of calcium homeostasis, on renal OPN expression. Three groups of rats were studied:1)acute(single injection of VitD, 200ng/100g BW s.c., 12h before sacrifice) and chronic VitD(daily injection of VitD 50ng/day/100g BW s.c. for 7d). 2) acute(single injection of PTH 50 microgram/100g BW i.p., 30min before sacrifice) and chronic PTH(infusion of PTH 6 microgram/day/100g BW s.c., for 7d via miniosmotic pump). 3) acute(single injection of calcitonin 25U/100g BW i.p., 1h before sacrifice) and chronic calcitonin(infusion of calcitonin 0.2U/ hr/kg BW s.c., for 7d via miniosmotic pump). Each of the study was compared with vehicle-treated animals. Kidneys were processed for immunohistochemistry and OPN expression was examined using a monoclonal antibody to OPN. In vehicle-treated animals, OPN was expressed primarily in DTL and PSE. In the acute VitD and PTH groups, OPN immunoreactivity appeared strongly in proximal tubule, and increased slightly in DTL and PSE. In the chronic VitD and PTH groups, there was a marked increase in OPN immunoreactivity in DTL, PSE, distal convoluted tubule(DCT) and thick ascending limb(TAL) of Henle's loop. Calcitonin groups showed no apparent change. In summary, this study demonstrates that OPN is constitutively expressed in the cells of the DTL and PSE, and induced in proximal tubule(PT), DCT and TAL by vitD and PTH. From these results we conclude that vitD and PTH play an important role in regulation of OPN expression not only in DTL and PSE but also in PT, DCT and TAL.
Animals ; Calcitonin* ; Calcium ; Epithelium ; Homeostasis ; Immunohistochemistry ; Kidney* ; Osteopontin* ; Rats* ; Vitamin D* ; Vitamins*

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

PURPOSE: Osteopontin is one of the major non-collagenous protein of hard tissue. Use of peptide domain of biologically active protein has some advantages. The objective of this experimental study is evaluation of periodontal regenerative potency of synthetic peptide gel which containing collagen binding domain of osteopontin in the degree III periodontal defect of beagle dogs. MATERIAL AND METHODS: Experimental degree III furcation defect was made in the mandibular third and fourth premolar of beagles. Regenerative material was applied during flap operation. 8 weeks after regenerative surgery, all animals were sacrificed and histomorphometric measurement was performed to calculate the linear percentage of the new cementum formation and the volume percentage of new bone formation. RESULT: The linear percent of new cementum formation was 41.6% at control group and 67.1% at test group and there was statistically significant difference. The volume percent of new bone formation was 52.1% at control group and 58.9% at test group. CONCLUSION: As the results of present experiment, synthetic peptide gel containing collagen binding domain of osteopontin significantly increase new bone and cementum formation in the degree III furcation defect of canine mandible.
Animals ; Bicuspid ; Collagen ; Dental Cementum ; Furcation Defects ; Mandible ; Osteogenesis ; Osteopontin ; Protein Structure, Tertiary ; Regeneration