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

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

PURPOSE: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-E1 osteoblastic cell line, particularly on the expression of osteocalcin and osteopontin. MATERIALS AND METHODS: Cells were irradiated with a single dose of 0.5, 1, 4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. After the specimens were harvested, RNA was extracted on the 3rd, 7th, 14th, and 21st day after irradiation. The RNA strands were reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR. RESULTS: The irradiated cells demonstrated a dose-dependent increase in osteocalcin and a dose-dependent decrease in osteopontin mRNA expression compared with the non-irradiated control group. The amount of osteocalcin mRNA expression decreased significantly at the 3rd day after irradiation of 0.5, 1, 4, and 8 Gy, and also decreased significantly at the 3rd, 14th, and 21st day after irradiation in the 8 Gy exposed group compared with the control group. The degree of osteopontin mRNA expression increased significantly at the 7th day after irradiation of 0.5, 1, 4, and 8 Gy. CONCLUSION: These results showed that each single dose of 0.5, 1, 4, and 8 Gy influenced the mRNA expression of osteocalcin and osteopontin associated with the calcification stage of osteoblastic cells, suggesting that each single dose affected bone formation at the cell level.
Cell Line* ; Cesium ; DNA, Complementary ; Osteoblasts* ; Osteocalcin* ; Osteogenesis ; Osteopontin* ; Polymerase Chain Reaction ; RNA ; 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 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: This study was undertaken to investigate the osteogenic induction potential of PGA & FBS mixture on a calvarial defect in the rabbit. METHODS: Twenty New zealand white rabbit, weighing from 3.5-4kg were allocated into each of the three groups. Four 8mm sized bone defects were made on the parietal bone by drilling. In group I, the bony defects were implanted with 50 micrometer thickness film containing mixture of PGA and FBS. In group II, with PGA only film, & in group III, the bony defects were left with no implants. Results were evaluated by using morphologic change, radiographic study, biochemical study and histologic examination at 1 week (group I n=7, group II n=7, group III n=14), 2 weeks (group I n=6, group II n=6, group III n=12) and 3 weeks (group I n=7, group II n=7, group III n=14) following implantation. RESULTS: In the morphologic & radiographic study, the formation and corticalization of callus were observed earlier in group I than in groups II and III (p<0.05). In histological examination, group I showed more abundant and faster new bone formation than in group II and III. In biochemical analysis, group I displayed more activity than in group II and III. Group I also showed more abundant osteopontin, osteocalcin than groups II and III. CONCLUSION: In conclusion, the results demonstrate that the mixture of PGA and FBS has an effect on osteoblastic formation in the rabbit model. It is considered that further evaluation of long term results on resorption, immunologic tissue reaction and response of applied mixture in the human model will be needed.
Bony Callus ; Humans ; New Zealand ; Osteoblasts ; Osteocalcin ; Osteogenesis ; Osteopontin ; Parietal Bone

PURPOSE: This study was undertaken to investigate the osteogenic induction potential of PGA & FBS mixture on a calvarial defect in the rabbit. METHODS: Twenty New zealand white rabbit, weighing from 3.5-4kg were allocated into each of the three groups. Four 8mm sized bone defects were made on the parietal bone by drilling. In group I, the bony defects were implanted with 50 micrometer thickness film containing mixture of PGA and FBS. In group II, with PGA only film, & in group III, the bony defects were left with no implants. Results were evaluated by using morphologic change, radiographic study, biochemical study and histologic examination at 1 week (group I n=7, group II n=7, group III n=14), 2 weeks (group I n=6, group II n=6, group III n=12) and 3 weeks (group I n=7, group II n=7, group III n=14) following implantation. RESULTS: In the morphologic & radiographic study, the formation and corticalization of callus were observed earlier in group I than in groups II and III (p<0.05). In histological examination, group I showed more abundant and faster new bone formation than in group II and III. In biochemical analysis, group I displayed more activity than in group II and III. Group I also showed more abundant osteopontin, osteocalcin than groups II and III. CONCLUSION: In conclusion, the results demonstrate that the mixture of PGA and FBS has an effect on osteoblastic formation in the rabbit model. It is considered that further evaluation of long term results on resorption, immunologic tissue reaction and response of applied mixture in the human model will be needed.
Bony Callus ; Humans ; New Zealand ; Osteoblasts ; Osteocalcin ; Osteogenesis ; Osteopontin ; Parietal Bone