• Journal of Periodontal and Implant Science
• /
• v.30 no.2
• /
• pp.389-405
• /
• 2000

The purpose of this study is to evaluate the effect of IGF-I for DNA synthetic activity and the mRNA expression of bone matrix protein, type I collagen and osteopontin in prolifetation and differentiation of MC3T3-E1 cells. To evaluate DNA synthetic activity, cells were seeded at $2{\times}10^4cells/ml$ in 24 well plates and to evaluate mRNA of type I collagen and osteopontin cells were seeded at $5{\times}10^5cells/ml$ in 100mm culture dishes. These cells were cultured in alpha-minimum essential medium(${\alpha}-MEM$) containing 10% fetal bovine serum at $37^{\circ}C$, 5% $CO_2$ incubator. For DNA synthetic activity test 1, 10, 100ng/ml IGF-I were added to the cells which had been cultured for 3 days before 24 hours. For type I collagen mRNA expression 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 10 days and for osteopontin mRNA expression 0.1, 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 15, 20 days. Cell proliferaton was measured by the incorporation of [$^3H$]-thymidine into DNA and expression for type I collagen and osteopontin were measured by northern blot analysis. The results were as follows : DNA synthetic activity were generally higher in experimental group than control group. Expressions of type I collagen mRNA were higher at 5 day group and much lower at 10 day group in the control groups. In the experimental groups, mRNA expressions were slightly increased when 1 ng/ml IGF-I were added to 5 day group and decreased in all experimental 10 day groups. Expressions of osteopontin mRNA were higher at 20 day groups and lower at 15 day groups than the control groups. In the experimental groups, mRNA expressions were incereased when 0.1, 1 ng/ml IGF-I were added to 5 day group and in all the 15 day groups, but decreased when 0.1, 1, 10 ng/ml IGF-I were added to 20 day groups. IGF-I stimulated DNA synthetic activity of MC3T3-E1 cells during proliferation stage significantly, did not greatly changed effects on type I collagen mRNA expression and stimulated osteopontin mRNA expression at 15 day especially. In conclusion, we suggests that IGF-I have a tendency of stimulation effect of DNA synthetic activity but do not stimulate type I collagen mRNA in proliferation stage of MC3T3-E1 cell cultures, and stimulate osteopontin mRNA in differentiation stage of MC3T3-E1 cell cultures.

• Journal of Nutrition and Health
• /
• v.49 no.1
• /
• pp.59-62
• /
• 2016

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 ($0-15{\mu}M$) 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.

• Proceedings of the Materials Research Society of Korea Conference
• /
• 2010.05a
• /
• pp.42.1-42.1
• /
• 2010

Poly-carprolactone (PCL)은 생분해성 고분자로 장기간의 임상실험 결과 생체에 독성이 없으며 생체친화성이 우수한 소재로 확인되어 PLGA, PLLA 등과 더불어 조직공학 분야에서 널리 사용되고 있는 생체재료이다. 그러나 PCL은 5개의 비극성 methylene group과 1개의 극성 ester group이 반복되는 지방족의 polyester로 구조상 탄소수가 많아 소수성을 띄는 단점을 가지고 있다. 이러한 표면이 소수성인 재료의 경우, 초기 단백질 흡착능이 떨어져 세포의 부착이 느린 속도로 일어나므로 세포 분화 및 조직 재생이 더디게 일어난다. 본 연구에서는 소수성의 PCL 표면의 단백질 흡착능을 증가시키기 위해 기능성 amine group을 부착하였으며, 또한 골재생을 촉진시킬 수 있는 세포외 기질인 collagen과 osteopontin을 부착함으로써 고기능성 PCL membrane을 제조하였다. 제조된 PCL membrane은 골재생용 조직공학에의 응용을 위해 지방유래 줄기세포를 이용하여 부착능 및 골세포로의 분화능을 확인하였다. 표면 성질의 변화에 의한 세포의 부착능의 변화를 confocal microscopy을 이용하여 부착에 관여하는 단백질의 발현을 확인하였으며, collagen과 osteopontin에 의한 골세포로의 분화능을 확인하기 위해 real time PCR을 통해 골세포의 분화 표지 유전자의 발현을 비교 분석하였다.

• Korean Journal of Veterinary Research
• /
• v.44 no.3
• /
• pp.335-341
• /
• 2004

This study was undertaken to examine the developmental expression of osteopontin(OPN) in the mouse brain. In Western blotting analysis, the expression of OPN was noted initially at embryonic stage and increased gradually after birth and decreased at postnatal day 60(P60). In immunohistochemistry, OPN expression was found in the interstitial nucleus Cajal and the substantia nigra reticularis in anterior part of the brain and in the inferior olivary complex, the parabrachial nucleus, the facial nucleus, the gigantocellular reticular nucleus, the trigeminal nucleus and the anterior interposed nucleus in posterior part of the brain at P31 and P60. In addition, OPN expression in widespread neurons appeared during the period of neuronal differentiation, increased just after birth and decreased with maturation. These results suggest that OPN contributes to developmental processes, including the differentiation and maturation of specific neuronal populations.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.12
• /
• pp.7315-7319
• /
• 2013

Background: The metastasis gene osteopontin (OPN) is subject to alternative splicing, which yields three messages, osteopontin-a, osteopontin-b and osteopontin-c. Osteopontin-c is selectively expressed in invasive, but not in noninvasive tumors. In the present study, we examined the expression of OPN-c in esophageal squamous cell carcinomas (ESCCs) and assessed its value as a diagnostic biomarker. Methods: OPN-c expression was assessed by immunohistochemistry in 63 ESCC samples and correlated with clinicopathologic factors. Expression was also examined in peripheral blood mononuclear cells (PBMCs) from 120 ESCC patients and 30 healthy subjects. The role of OPN-c mRNA as a tumor marker was investigated by receiver operating characteristic curve (ROC) analysis. Results: Immunohistochemistry showed that OPN-c was expressed in 30 of 63 cancer lesions (48%)and significantly associated with pathological T stage (P=0.038) and overall stage (P=0.023). Real time PCR showed that OPN-c mRNA was expressed at higher levels in the PBMCs of ESCC patients than in those of healthy subjects (P<0.0001) with a sensitivity as an ESCC biomarker of 86.7%. Conclusion: Our findings suggest that expression of OPN-c is significantly elevated in ESCCs and this upregulation could be a potential diagnostic marker.

• Journal of Animal Science and Technology
• /
• v.45 no.3
• /
• pp.491-498
• /
• 2003

The purpose of this study is to observe purification and properties of osteopontin(OPN) from bovine milk. The purification of osteopontin from bovine milk was performed by using ion-exchange and hydrophobic chromatography. SDS-PAGE analysis revealed that the protein migrated at Mw. 60,000. NH2-terminal sequence analysis of the first seven amio acids revealed the protein to be identical to that previously reported for bovine OPN. 35-wk-old chickens, including 3 Single Comb White Leghorn (SCWL), were used to produce egg yolk antibody(IgY) against OPNas a antigen. However, the anti-OPN antibody activities determined by ELISA. Immunological assy of OPN in milk was performed using radial immunodiffusion test based on the standard curve of pure OPN. The radial precipitation lines of four different milk samples indicated that the concentrations of OPN in the milk samples were within the range of 31.7 to 39.7${\mu}g$/ml. On inhibition with OPN on precipitation of calcium phosphate, OPN was slightly higher than casein phosphopeptide(CPP) and poly-glutamic acid.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.4
• /
• pp.652-656
• /
• 2007

Hydroxyapatite [Ca10(PO4)6(OH)2, HAp] and titanium(Ti) metal are known to be excellent materials with high affinities for natural bone through the apatite. Tricalcium phosphate [Ca3(PO4)2, TCP] is a promising alternative material because it is similar to HAp in its physical properties and biocompatibility. To examine the influence of hydroxyapatite, tricalcium phosphate, pure titanium and pre-treated titanium on osteopontin expression in osteoblasts, RNA was extracted from proliferated and cultured osteoblast cells and OPN mRNA expression was observed by RT-PCR.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.26 no.4
• /
• pp.652-663
• /
• 1999

Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of the interactions between different tissues within the cranial sutures. Interestingly, point mutaions in the genes encoding for the fibroblast growth factor receptors(FGFRs), especially FGFR2, cause various types of human craniosynostosis syndromes. To elucidate the function of these genes in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of FGFR2(BEK) and osteopontin, an early marker of osteogenic differentiation, in the sagittal suture of calvaria during embryonic(E15-E18) and postnatal stage(P1-P3). FGFR2(BEK) was intensely expressed in the osteogenic fronts, whose cells undergo differentiation into osteoprogenitor cells that ultimately lay down the bone matrix. Osteopontin was expressed throughout the parietal bones excluding the osteogenic fronts, the periphery of the parietal bones. To further examine the role of FGF-mediated FGFR signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of FGF2 soaked beads onto both the osteogenic fronts and mid-mesenchyme of sagittal suture after 36 hours organ culture resulted in the increase of the tissue thickness and cell number around FGF2 beads, moreover FGF4-soaked beads implanted onto the osteogenic fronts stimulated suture closure due to an accelerated bone growth, compared to FGF4 beads placed onto mid-mesenchyme of sagittal suture and BSA control beads. In addition FGF2 induced the ectopic expression of osteopontin and Msx1 genes. Taken together, these data indicate that FGF-mediated FGFR signaling has a important role in regulating the cranial bone growth and maintenance of cranial suture, and suggest that FGF-mediated FGFR signaling is involved in regulating the balance between the cell proliferation and differentiation through inducing the expression of osteopontin and Msx1 genes.

• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.142.2-142.2
• /
• 2003

The activation of the hepatic stellate cell (HSC) is a key step in liver fibrogenesis. We investigated the changes of global gene expression during activation in hepatic stellate cells using a rat cDNA microarray with 5, 000 sequence-verified clones. We identified osteopontin (OPN), a secreted matrix protein, as one of the upregulated factors. Northern analysis showed OPN mRNA was increasingly expressed during progressive activation of cultured rat HSCs and in models of experimental liver fibrosis. (omitted)

• Journal of Periodontal and Implant Science
• /
• v.38 no.4
• /
• pp.621-628
• /
• 2008

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.