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

• Preventive Nutrition and Food Science
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• v.18 no.2
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• pp.92-97
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• 2013

The calcification of vascular smooth muscle cells (VSMCs) is considered one of the major contributors for vascular disease. Phosphate is known as the inducer for VSMC calcification. In this study, we assessed whether phosphate affected cell viability and fetuin-A, a calcification inhibitor protein, both which are related to VSMC calcification. Also, VSMC viability by zinc level was assessed. The results showed that phosphate increased Ca and P deposition in VSMCs (A7r5 cell line, rat aorta origin). This phosphate-induced Ca and P deposition was consistent with the decreased A7r5 cell viability (P<0.05), which implies phosphate-induced calcification in A7r5 cells might be due to the decreased VSMC cell viability. As phosphate increased, the protein expression of fetuin-A protein was up-regulated. A7r5 cell viability decreased as the addition of cellular zinc level was decreased (P<0.05). The results suggested that zinc deficiency causes the decreased cell viability and it would be the future study to clarify how zinc does act for VSMC cell viability. The results suggest that the decreased VSMC viability by high P or low Zn in VSMCs may be the risk factor for vascular disease.

• Fisheries and Aquatic Sciences
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• v.17 no.2
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• pp.269-273
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• 2014

Coralline algae, the algal whitening phenomenon-causing seaweeds, are characterized by calcareous deposits in the cell wall. The viability of the coralline algae Lithophyllum yessoense and Corallina pilulifera was quantitated using a triphenyltetrazolium chloride assay and eight calcification inhibitors. Among these inhibitors, ferric citrate showed the strongest inhibition of coralline algae viability. The concentrations of ferric citrate conferring 50% inhibition were 1.7 and 3.8 mM for L. yessoense and C. pilulifera, respectively. Thus, at a specific concentration and in a localized area, ferric citrate may be used to prevent the blooming of coralline algae.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.2
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• pp.111-118
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• 2015

Osteoprotegerin (OPG), receptor activator of NF-${\kappa}B$ ligand (RANKL)/receptor activator of NF-${\kappa}B$ (RANK) axis, and TNF-related apoptosis-inducing ligand (TRAIL) participate in vascular calcification process including atherosclerosis, but their contributions under high glucose (HG) and phosphate (HP) condition for a long-term period (more than 2 weeks) have not been fully determined. In this study, we evaluated the effects of HG and HP levels over 2 or 4 weeks on the progression of vascular calcification in rat vascular smooth muscle cells (VSMCs). Calcium deposition in VSMCs was increased in medium containing HG (30 mmol/L D-glucose) with ${\beta}$-glycerophosphate (${\beta}$-GP, 12 mmol/L) after 2 weeks and increased further after 4 weeks. OPG mRNA and protein expressions were unchanged in HG group with or without ${\beta}$-GP after 2 weeks. However, after 4 weeks, OPG mRNA and protein expressions were significantly lower in HG group with ${\beta}$-GP. No significant expression changes were observed in RANKL, RANK, or TRAIL during the experiment. After 4 weeks of treatment in HG group containing ${\beta}$-GP and rhBMP-7, an inhibitor of vascular calcification, OPG expressions were maintained. Furthermore, mRNA expression of alkaline phosphatase (ALP), a marker of vascular mineralization, was lower in the presence of rhBMP-7. These results suggest that low OPG levels after long term HG and phosphate stimulation might reduce the binding of OPG to RANKL and TRAIL, and these changes could increase osteo-inductive VSMC differentiation, especially vascular mineralization reflected by increased ALP activity during vascular calcification.

• International Journal of Oral Biology
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• v.33 no.1
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• pp.33-43
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• 2008

In the process of bone remodeling, mineral phase of bone is dissolved by osteoclasts, resulting in elevation of calcium concentration in micro-environment. This study was performed to explore the effect of high extracellular calcium ($Ca{^{2+}}_e$) on mineralized nodule formation and on the expression of progressive ankylosis (Ank), plasma cell membrane glycoprotein-1 (PC-1) and osteopontin by primary cultured mouse calvarial cells. Osteoblastic differentiation and mineralized nodule formation was induced by culture of mouse calvarial cells in osteoblast differentiation medium containing ascorbic acid and ${\beta}$-glycerophosphate. Although Ank, PC-1 and osteopontin are well known inhibitors of mineralization, expression of these genes were induced at the later stage of osteoblast differentiation during when expression of osteocalcin, a late marker gene of osteoblast differentiation, was induced and mineralization was actively progressing. High $Ca{^{2+}}_e$(10 mM) treatment highly enhanced mRNA expression of Ank, PC-1 and osteopontin in the late stage of osteoblast differentiation but not in the early stage. Inhibition of p44/42 MAPK activation but not that of protein kinase C suppressed high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin. When high $Ca{^{2+}}_e$(5 mM or 10 mM) was present in culture medium during when mineral deposition was actively progressing, matrix calcifiation was significantly increased by high $Ca{^{2+}}_e$. This stimulatory effect was abolished by pyrophosphate (5 mM) or levamisole (0.1-0.5 mM), an alkaline phosphatase inhibitor. In addition, probenecid (2mM), an inhibitor of Ank, suppressed matrix calcification in both control and high $Ca{^{2+}}_{e^-}$treated group, suggesting the possible role of Ank in matrix calcification by osteoblasts. Taken together, these results showed that high $Ca{^{2+}}_e$ stimulates expression of Ank, PC-1 and osteopontin as well as matrix calcification in late differentiation stage of osteoblasts and that p44/42 MAPK activation is involved in high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.18-24
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• 2002

The present study was performed to investigate the excellent microbial anticaries substance, aminoglycoside antibiotic, which is more effective than chlorhexidine for the treatment of dental caries. The aminoglycoside antibiotic against Streptococcus mutans JC-2 from a novel alkaliphilic Bacillus alkalophilshaggy JY-827 exhibited no significant difference at the treatment concentration of $2.5{\times}10^{-7}M$, however, it inhibited the activity of the Streptococcus mutans glucosyltransferase by 70.2% and 99.8% at the concentrations of $2.5{\times}10^{-7}$M\;and\;2.5{\times}10^{-6}M$, respectively. Lineweaver-Burk plot of the inhibitory aminoglycoside antibiotic showed competitive inhibition, with $K_i$ value of $6.4{\times}10^{-6}$ M. The aminoglycoside antibiotic did not show any cytotoxicity against human gingival cells. To evaluate the industrial applicability of the aminoglycoside antibiotic, a toothpaste containing this substance was prepared and tested on the extracted human teeth. The inhibitory rate of tooth calcification and calcium ion elution by the aminoglycoside antibiotic were 50% and 2.5 times, respectively. These results suggested that the aminoglycoside antibiotic from Bacillus alkalophilshaggy JY-827 is an effective agent against dental caries.

• Restorative Dentistry and Endodontics
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• v.25 no.4
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• pp.509-526
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• 2000

It is known that injuries to the dentin have a corresponding inflammatory effect on the pulp and these inflammatory effects frequently result in pulpal pathoses due to progressive degradation of pulpal connective tissue. It was supposed that the tissue degradation in different inflammatory process was controlled by proteinase activity and antiproteinase activity. Therefore, the purpose of this study was to examine the pulp and periapical pathoses in terms of the activities of proteinase and proteinase inhibitor, 37 pulpal tissues were divided by clinical diagnostic criteria into normal pulp, acute inflamed pulp, and chronic inflamed pulp, and then those groups were subdivided by histopathological findings into 5 pulpal pathoses groups, i.e. normal pulp (P1, n=8), chronic pulpitis with fibrotic change (P2, n=2), chronic pulpitis with dystrophic calcification (P3, n=11), chronic pulpitis with pulp abscess (P4, n=7), acute pulpitis with necrotic change (P5, n=4), 26 periapical tissues were also divided by ordinary histopathological findings into 3 periapical pathoses group, i.e., granuloma (A1, n=17), cyst (A2, n=2) and abscess (A3, n=7). The activities of proteinases (cathepsin G, MMP-3) and proteinase inhibitors (${\alpha}1$-AT, TIMP-1 and, SLPI) were evaluated by RT-PCR and immunohistochemical methods. The results were as follows. 1. Generally, the intensity of immunohistochemical staining of proteinases and proteinase inhibitors increased in P2 and P5 groups compared to P1 group. 2. The immunohistochemical stain of proteinases and proteinase inhibitors was intensely detected in P2 group, showing low inflammatory reaction and low tissue degradation, but it was reduced in P3 and P4 groups, showing severe tissue degradation. 3. The distribution of proteinases and proteinase inhibitors in pulpal pathoses was consistently presented by immunohistochemical staining, while the expression of proteinase and/or proteinase inhibitors mRNAs in pulpal pathoses was occasionally detected by RT-PCR methods. 4. RT-PCR of proteinase and proteinase inhibitors was usually positive in P2, showing rare tissue degradation, but it was almost negative in P3 and P4, showing severe tissue degradation. 5. We presume that the reason why the level of proteinase and proteinase inhibitors was so sparse in RT-PCR method is due to the abrupt decrease of mRNA synthesis or degradation of synthesized mRNA of proteinase and/or proteinase inhibitors depend on the inflammatory reaction and/or on the degradation of pulp tissues(P3, P4). 6. Pulpal pathoses groups showed significant lower RT-PCR detection of proteinases and proteinase inhibitors than the periapical pathoses group(p<0.05), and there is no significant difference among the periapical pathoses groups(p>0.05).