Effects of 1,25-dihydroxyvitamin D3 on the differentiation of MC3T3-E1 osteoblast-like cells
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Purpose: The purpose of this study was to evaluate the effects of 1,25-dihydroxyvitamin D3 on the proliferation, differentiation, and matrix mineralization of MC3T3-E1 osteoblast-like cells in vitro. Methods: MC3T3-E1 osteoblastic cells and 1,25-dihydroxyvitamin D3 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 D3 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 D3 (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 D3 (10−12 M). Alizarin red S staining after treatment with 1,25-dihydroxyvitamin D3 (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 D3 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.
Abstract
Purpose: The purpose of this study was to evaluate the effects of 1,25-dihydroxyvitamin D3 on the proliferation, differentiation, and matrix mineralization of MC3T3-E1 osteoblast-like cells in vitro. Methods: MC3T3-E1 osteoblastic cells and 1,25-dihydroxyvitamin D3 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 D3 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 D3 (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 D3 (10−12 M). Alizarin red S staining after treatment with 1,25-dihydroxyvitamin D3 (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 D3 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.
참고문헌 (33)
- Schropp L, "Bone healing and soft tissue contour changes following single-tooth extraction: a clinical and radiographic 12-month prospective study", Int J Periodontics Restorative Dent, 23, pp.313~323 (2003)
- Hynes K, "Clinical utility of stem cells for periodontal regeneration", Periodontol 2000, 59, pp.203~227 (2012)[DOI]
- Rosenberg E, "Biologic and clinical considerations for autografts and allografts in periodontal regeneration therapy", Dent Clin North Am, 42, pp.467~490 (1998)
- Fiorellini JP, "Osseointegration of titanium implants following guided bone regeneration using expanded polytetrafluoroethylene membrane and various bone fillers", Int J Periodontics Restorative Dent, 27, pp.287~294 (2007)
- Nakashima M, "Bone morphogenetic proteins in dentin regeneration for potential use in endodontic therapy", Cytokine Growth Factor Rev, 16, pp.369~376 (2005)[DOI]
- Nakashima M, "The application of bone morphogenetic proteins to dental tissue engineering", Nat Biotechnol, 21, pp.1025~1032 (2003)[DOI]
- Srisuwan T, "Molecular aspects of tissue engineering in the dental field", Periodontol 2000, 41, pp.88~108 (2006)[DOI]
- Anusaksathien O, "Growth factor delivery to re-engineer periodontal tissues", Curr Pharm Biotechnol, 3, pp.129~139 (2002)[DOI]
- Minghetti PP, "1,25-dihydroxy-vitamin D3 receptors: gene regulation and genetic circuitry", FASEB J, 2, pp.3043~3053 (1988)[DOI]
- Henry HL, "Studies on the mechanism of action of calciferol VII. Localization of 1,25-dihydroxy-vitamin D3 in chick parathyroid glands", Biochem Biophys Res Commun, 62, pp.781~788 (1975)[DOI]
- Minghetti PP, "1,25(OH)2-vitamin D3 receptors: gene regulation and genetic circuitry", FASEB J, 2, pp.3043~3053 (1988)[DOI]
- Norman AW, "Vitamin D: the calcium homeostatic steroid hormone" (1979), Academic Press
- Norman AW, "The vitamin D endocrine system: steroid metabolism, hormone receptors, and biological response (calcium binding proteins)", Endocr Rev, 3, pp.331~366 (1982)[DOI]
- Haneji T, "<TEX>$1{\alpha}$<", TEX>,25-Dihydroxyvitamin D3 and analogues of vitamin D3 induce alkaline phosphatase activity in osteoblastic cells derived from newborn mouse calvaria, 1983, pp.94, 1127~1132
- Matsumoto T, "Stimulation by 1, 25-dihydroxyvitamin D3 of in vitro mineralization induced by osteoblast-like MC3T3-E1 cells", Bone, 12, pp.27~32 (1991)[DOI]
- Seibert J, "Localized ridge augmentation in dogs: a pilot study using membranes and hydroxyapatite", J Periodontol, 61, pp.157~165 (1990)[DOI]
- Petite H, "Tissue-engineered bone regeneration", Nat Biotechnol, 18, pp.959~963 (2000)[DOI]
- Choi JY, "Expression patterns of bone related proteins during osteoblastic differentiation in MC3T3E1 cells", J Cell Biochem, 61, pp.609~618 (1996)[DOI]
- Lian JB, "Primer on the metabolic bone diseases and disorders of mineral metabolism", pp.14~19 (1999), Lippincott Williams & Wilkins
- Ming-Zhen Zheng, "Effect of Cornus Officinalis Extract on the Differentiation of MC3T3-E1 Osteoblast-like Cells", 조직공학과 재생의학, 12(4), pp.113~121 (2015), 한국조직공학과 재생의학회
- Spelsberg TC, "The actions and interactions of sex steroids and growth factors", cytokines on the skeleton, 1999, pp.13, 819~828
- Fraser JD, "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, 263, pp.911~916 (1988)
- Manolagas SC, "1,25-Dihydroxyvitamin D3 stimulates the alkaline phosphatase activity of osteoblast-like cells", J Biol Chem, 256, pp.7115~7117 (1981)
- Price PA, "1,25-Dihydroxyvitamin D3 increases synthesis of the vitamin K-dependent bone protein by osteosarcoma cells", J Biol Chem, 255, pp.11660~11663 (1980)
- Rowe DW, "Regulation of collagen synthesis in fetal rat calvaria by 1,25-dihydroxyvitamin D3", J Biol Chem, 257, pp.8009~8015 (1982)
- Haussler MR, "Molecular mechanisms of vitamin D action", Calcif Tissue Int, 92, pp.77~98 (2013)[DOI]
- Carlberg C, "Vitamin D receptor signaling mechanisms: integrated actions of a well-defined transcription factor", Steroids, 78, pp.127~136 (2013)[DOI]
- Franceschi RT, "Transcriptional regulation of osteoblasts", Cells Tissues Organs, 189, pp.144~152 (2009)[DOI]
- Jones G, "Pharmacokinetics of vitamin D toxicity", Am J Clin Nutr, 88, pp.582S~586S (2008)[DOI]
- Sigurdsson TJ, "Periodontal repair in dogs: evaluation of rhBMP-2 carriers", Int J Periodontics Restorative Dent, 16, pp.524~537 (1996)
- Kurihara N, "Effects of 1,25-dihydroxyvitamin D3 on osteoblastic MC3T3-E1 cells", Endocrinology, 118, pp.940~947 (1986)[DOI]
- 이승용, "홍화씨 추출물이 MC3T3-E1 조골세포의 증식과 분화에 미치는 영향", Oral Biology Research, 37(2), pp.73~81 (2013), 치의학연구원[DOI]
- Wang W, "Sanguis Draconis resin stimulates osteoblast alkaline phosphatase activity and mineralization in MC3T3-E1 cells", J Ethnopharmacol, 142, pp.168~174 (2012)[DOI]