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http://dx.doi.org/10.5051/jpis.2018.48.1.34

Effects of 1,25-dihydroxyvitamin D3 on the differentiation of MC3T3-E1 osteoblast-like cells  

Kim, Hyun-Soo (Department of Periodontology, Chosun University School of Dentistry)
Zheng, Mingzhen (Department of Stomatology, Affiliated Hospital of Yanbian University)
Kim, Do-Kyung (Department of Oral Physiology, Chosun University School of Dentistry)
Lee, Won-Pyo (Department of Periodontology, Chosun University School of Dentistry)
Yu, Sang-Joun (Department of Periodontology, Chosun University School of Dentistry)
Kim, Byung-Ock (Department of Periodontology, Chosun University School of Dentistry)
Publication Information
Journal of Periodontal and Implant Science / v.48, no.1, 2018 , pp. 34-46 More about this Journal
Abstract
Purpose: The purpose of this study was to evaluate the effects of 1,25-dihydroxyvitamin $D_3$ on the proliferation, differentiation, and matrix mineralization of MC3T3-E1 osteoblast-like cells in vitro. Methods: MC3T3-E1 osteoblastic cells and 1,25-dihydroxyvitamin $D_3$ were prepared. Cytotoxic effects and osteogenic differentiation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity assay, ALP staining, alizarin red S staining, and reverse transcription-polymerase chain reaction (RT-PCR) for osteogenic differentiation markers such as ALP, collagen type I (Col-I), osteocalcin (OCN), vitamin D receptor (VDR), and glyceraldehyde 3-phosphate dehydrogenase. Results: The MTT assay showed that 1,25-dihydroxyvitamin $D_3$ did not inhibit cell growth and that the rate of cell proliferation was higher than in the positive control group at all concentrations. ALP activity was also higher than in the positive control group at low concentrations of 1,25-dihydroxyvitamin $D_3$ ($10^{-10}$, $10^{-12}$, and $10^{-14}M$). RT-PCR showed that the gene expression levels of ALP, Col-I, OCN, and vitamin D receptor (VDR) were higher at a low concentration of 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$). Alizarin red S staining after treatment with 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$) showed no significant differences in the overall degree of calcification. In contrast to the positive control group, formation of bone nodules was induced in the early stages of cell differentiation. Conclusions: We suggest that 1,25-dihydroxyvitamin $D_3$ positively affects cell differentiation and matrix mineralization. Therefore, it may function as a stimulating factor in osteoblastic bone formation and can be used as an additive in bone regeneration treatment.
Keywords
Bone regeneration; Calcitriol; Osteoblasts;
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1 Schropp L, Wenzel A, Kostopoulos L, Karring T. Bone healing and soft tissue contour changes following single-tooth extraction: a clinical and radiographic 12-month prospective study. Int J Periodontics Restorative Dent 2003;23:313-23.
2 Hynes K, Menicanin D, Gronthos S, Bartold PM. Clinical utility of stem cells for periodontal regeneration. Periodontol 2000 2012;59:203-27.   DOI
3 Rosenberg E, Rose LF. Biologic and clinical considerations for autografts and allografts in periodontal regeneration therapy. Dent Clin North Am 1998;42:467-90.
4 Fiorellini JP, Kim DM, Nakajima Y, Weber HP. Osseointegration of titanium implants following guided bone regeneration using expanded polytetrafluoroethylene membrane and various bone fillers. Int J Periodontics Restorative Dent 2007;27:287-94.
5 Nakashima M. Bone morphogenetic proteins in dentin regeneration for potential use in endodontic therapy. Cytokine Growth Factor Rev 2005;16:369-76.   DOI
6 Nakashima M, Reddi AH. The application of bone morphogenetic proteins to dental tissue engineering. Nat Biotechnol 2003;21:1025-32.   DOI
7 Franceschi RT, Ge C, Xiao G, Roca H, Jiang D. Transcriptional regulation of osteoblasts. Cells Tissues Organs 2009;189:144-52.   DOI
8 Rowe DW, Kream BE. Regulation of collagen synthesis in fetal rat calvaria by 1,25-dihydroxyvitamin D3. J Biol Chem 1982;257:8009-15.
9 Haussler MR, Whitfield GK, Kaneko I, Haussler CA, Hsieh D, Hsieh JC. Molecular mechanisms of vitamin D action. Calcif Tissue Int 2013;92:77-98.   DOI
10 Carlberg C, Campbell MJ. Vitamin D receptor signaling mechanisms: integrated actions of a well-defined transcription factor. Steroids 2013;78:127-36.   DOI
11 Jones G. Pharmacokinetics of vitamin D toxicity. Am J Clin Nutr 2008;88:582S-586S.   DOI
12 Sigurdsson TJ, Nygaard L, Tatakis DN, Fu E, Turek TJ, Jin L, et al. Periodontal repair in dogs: evaluation of rhBMP-2 carriers. Int J Periodontics Restorative Dent 1996;16:524-37.
13 Kurihara N, Ishizuka S, Kiyoki M, Haketa Y, Ikeda K, Kumegawa M. Effects of 1,25-dihydroxyvitamin D3 on osteoblastic MC3T3-E1 cells. Endocrinology 1986;118:940-7.   DOI
14 Lee SY, Kim SG, Lee SA, Park MG, Park SY, Oh JS, et al. Influence of Carthamus tinctorius seed extract on proliferation and differentiation of MC3T3-E1 osteoblast cells. Oral Biol Res 2013;37:73-81.   DOI
15 Henry HL, Norman AW. Studies on the mechanism of action of calciferol VII. Localization of 1,25-dihydroxy-vitamin D3 in chick parathyroid glands. Biochem Biophys Res Commun 1975;62:781-8.   DOI
16 Srisuwan T, Tilkorn DJ, Wilson JL, Morrison WA, Messer HM, Thompson EW, et al. Molecular aspects of tissue engineering in the dental field. Periodontol 2000 2006;41:88-108.   DOI
17 Anusaksathien O, Giannobile WV. Growth factor delivery to re-engineer periodontal tissues. Curr Pharm Biotechnol 2002;3:129-39.   DOI
18 Minghetti PP, Norman AW. 1,25-dihydroxy-vitamin D3 receptors: gene regulation and genetic circuitry. FASEB J 1988;2:3043-53.   DOI
19 Minghetti PP, Norman AW. 1,25(OH)2-vitamin D3 receptors: gene regulation and genetic circuitry. FASEB J 1988;2:3043-53.   DOI
20 Norman AW. Vitamin D: the calcium homeostatic steroid hormone. New York (NY): Academic Press; 1979.
21 Norman AW, Roth J, Orci L. The vitamin D endocrine system: steroid metabolism, hormone receptors, and biological response (calcium binding proteins). Endocr Rev 1982;3:331-66.   DOI
22 Haneji T, Kurihara N, Ikeda K, Kumegawa M. $1{\alpha}$,25-Dihydroxyvitamin D3 and analogues of vitamin D3 induce alkaline phosphatase activity in osteoblastic cells derived from newborn mouse calvaria. J Biochem 1983;94:1127-32.   DOI
23 Choi JY, Lee BH, Song KB, Park RW, Kim IS, Sohn KY, et al. Expression patterns of bone related proteins during osteoblastic differentiation in MC3T3E1 cells. J Cell Biochem 1996;61:609-18.   DOI
24 Matsumoto T, Igarashi C, Takeuchi Y, Harada S, Kikuchi T, Yamato H, et al. Stimulation by 1, 25-dihydroxyvitamin D3 of in vitro mineralization induced by osteoblast-like MC3T3-E1 cells. Bone 1991;12:27-32.   DOI
25 Seibert J, Nyman S. Localized ridge augmentation in dogs: a pilot study using membranes and hydroxyapatite. J Periodontol 1990;61:157-65.   DOI
26 Petite H, Viateau V, Bensaid W, Meunier A, de Pollak C, Bourguignon M, et al. Tissue-engineered bone regeneration. Nat Biotechnol 2000;18:959-63.   DOI
27 Lian JB, Stein GS, Canalis E, Robey PG, Boskey AL. Bone formation: osteoblast lineage cells, growth factors, matrix proteins and the mineralization process. In: Favus M, editor. Primer on the metabolic bone diseases and disorders of mineral metabolism. 4th ed. Philadelphia (PA): Lippincott Williams & Wilkins; 1999. p.14-9.
28 Zheng MZ, Lee SY, Yu SJ, Kim BO. Effect of Cornus officinalis extract on the differentiation of MC3T3-E1 osteoblast-like cells. Tissue Eng Regen Med 2015;12:113-21.
29 Spelsberg TC, Subramaniam M, Riggs BL, Khosla S. The actions and interactions of sex steroids and growth factors/cytokines on the skeleton. Mol Endocrinol 1999;13:819-28.   DOI
30 Fraser JD, Otawara Y, Price PA. 1,25-Dihydroxyvitamin D3 stimulates the synthesis of matrix gammacarboxyglutamic acid protein by osteosarcoma cells. Mutually exclusive expression of vitamin K-dependent bone proteins by clonal osteoblastic cell lines. J Biol Chem 1988;263:911-6.
31 Manolagas SC, Burton DW, Deftos LJ. 1,25-Dihydroxyvitamin D3 stimulates the alkaline phosphatase activity of osteoblast-like cells. J Biol Chem 1981;256:7115-7.
32 Wang W, Olson D, Cheng B, Guo X, Wang K. Sanguis Draconis resin stimulates osteoblast alkaline phosphatase activity and mineralization in MC3T3-E1 cells. J Ethnopharmacol 2012;142:168-74.   DOI
33 Price PA, Baukol SA. 1,25-Dihydroxyvitamin D3 increases synthesis of the vitamin K-dependent bone protein by osteosarcoma cells. J Biol Chem 1980;255:11660-3.