The effects of bone morphogenetic protein-2 and enamel matrix derivative on the bioactivity of mineral trioxide aggregate in MC3T3-E1cells
1
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
Objectives: The effects of bone morphogenetic protein-2 (BMP-2) and enamel matrix derivative (EMD) respectively with mineral trioxide aggregate (MTA) on hard tissue regeneration have been investigated in previous studies. This study aimed to compare the osteogenic effects of MTA/BMP-2 and MTA/EMD treatment in MC3T3-E1 cells. Materials and Methods: MC3T3-E1 cells were treated with MTA (ProRoot, Dentsply), BMP-2 (R&D Systems), EMD (Emdogain, Straumann) separately and MTA/BMP-2 or MTA/ EMD combination. Mineralization was evaluated by staining the calcium deposits with alkaline phosphatase (ALP, Sigma-Aldrich) and Alizarin red (Sigma-Aldrich). The effects on the osteoblast differentiation were evaluated by the expressions of osteogenic markers, including ALP, bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN) and osteonectin (OSN), as determined by reverse-transcription polymerase chain reaction analysis (RT-PCR, AccuPower PCR, Bioneer). Results: Mineralization increased in the BMP-2 and MTA/BMP-2 groups and increased to a lesser extent in the MTA/ EMD group but appeared to decrease in the MTA-only group based on Alizarin red staining. ALP expression largely decreased in the EMD and MTA/EMD groups based on ALP staining. In the MTA/BMP-2 group, mRNA expression of OPN on day 3 and BSP and OCN on day 7 significantly increased. In the MTA/EMD group, OSN and OCN gene expression significantly increased on day 7, whereas ALP expression decreased on days 3 and 7 (p < 0.05). Conclusions: These results suggest the MTA/BMP-2 combination promoted more rapid differentiation in MC3T3-E1 cells than did MTA/EMD during the early mineralization period.
Abstract
Objectives: The effects of bone morphogenetic protein-2 (BMP-2) and enamel matrix derivative (EMD) respectively with mineral trioxide aggregate (MTA) on hard tissue regeneration have been investigated in previous studies. This study aimed to compare the osteogenic effects of MTA/BMP-2 and MTA/EMD treatment in MC3T3-E1 cells. Materials and Methods: MC3T3-E1 cells were treated with MTA (ProRoot, Dentsply), BMP-2 (R&D Systems), EMD (Emdogain, Straumann) separately and MTA/BMP-2 or MTA/ EMD combination. Mineralization was evaluated by staining the calcium deposits with alkaline phosphatase (ALP, Sigma-Aldrich) and Alizarin red (Sigma-Aldrich). The effects on the osteoblast differentiation were evaluated by the expressions of osteogenic markers, including ALP, bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN) and osteonectin (OSN), as determined by reverse-transcription polymerase chain reaction analysis (RT-PCR, AccuPower PCR, Bioneer). Results: Mineralization increased in the BMP-2 and MTA/BMP-2 groups and increased to a lesser extent in the MTA/ EMD group but appeared to decrease in the MTA-only group based on Alizarin red staining. ALP expression largely decreased in the EMD and MTA/EMD groups based on ALP staining. In the MTA/BMP-2 group, mRNA expression of OPN on day 3 and BSP and OCN on day 7 significantly increased. In the MTA/EMD group, OSN and OCN gene expression significantly increased on day 7, whereas ALP expression decreased on days 3 and 7 (p < 0.05). Conclusions: These results suggest the MTA/BMP-2 combination promoted more rapid differentiation in MC3T3-E1 cells than did MTA/EMD during the early mineralization period.
참고문헌 (34)
- Bashutski JD, "Periodontal and endodontic regeneration", J Endod, 35, pp.321~328 (2009)
- Schwartz RS, "Mineral trioxide aggregate: a new material for endodontics", J Am Dent Assoc, 130, pp.967~975 (1999)
- Torabinejad M, "Histologic assessment of mineral trioxide aggregate as a root-end filling in monkeys", J Endod, 23, pp.225~228 (1997)
- Parirokh M, "Mineral trioxide aggregate: a comprehensive literature review-part III: clinical applications, drawbacks, and mechanism of action", J Endod, 36, pp.400~413 (2010)
- Min KS, "The combined effect of mineral trioxide aggregate and enamel matrix derivative on odontoblastic differentiation in human dental pulp cells", J Endod, 35, pp.847~851 (2009)
- Chen D, "Bone morphogenetic proteins", Growth Factors, 22, pp.233~241 (2004)
- Yamaguchi A, "Recombinant human bone morphogenetic protein-2 stimulates osteoblastic maturation and inhibits myogenic differentiation in vitro", J Cell Biol, 113, pp.681~687 (1991)
- Welch RD, "Effect of recombinant human bone morphogenetic protein-2 on fracture healing in a goat tibial fracture model", J Bone Miner Res, 13, pp.1483~1490 (1998)
- Hogan BL., "Bone morphogenetic proteins in development", Curr Opin Genet Dev, 6, pp.432~438 (1996)
- Ko H, "Cytotoxicity of mineral trioxide aggregate (MTA) and bone morphogenetic protein 2 (BMP-2) and response of rat pulp to MTA and BMP-2.", Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 109, pp.e103~e108 (2010)
- Nakashima M., "Induction of dentin formation on canine amputated pulp by recombinant human bone morphogenetic proteins (BMP)-2 and -4", J Dent Res, 73, pp.1515~1522 (1994)
- Hammarström L., "Enamel matrix, cementum development and regeneration", J Clin Periodontol, 24, pp.658~668 (1997)
- Jiang J, "Enamel matrix derivative prolongs primary osteoblast growth", J Endod, 27, pp.110~112 (2001)
- Chung H, "Comparison of the effects of enamel matrix derivative and mineral trioxide aggregate on the mineralization potential of human cementum-derived cells", J Dent Sci, 6, pp.153~157 (2011)
- Wozney JM., "The bone morphogenetic protein family: multifunctional cellular regulators in the embryo and adult", Eur J Oral Sci, 106(Supplement 1), pp.160~166 (1998)
- Lianjia Y, "Immunohistochemical localization of bone morphogenetic protein (BMP) in calcifying fibrous epulis", J Oral Pathol Med, 22, pp.406~410 (1993)
- Lee SY, "Effects of Simvastain and Enamel Matrix Derivative on Portland Cement with Bismuth Oxide-induced Growth and Odontoblastic Differentiation in Human Dental Pulp Cells", J Endod, 38(3), pp.405~410 (2012), ELSEVIER SCIENCE INC
- Palioto DB, "Effects of enamel matrix derivative and transforming growth factor-ββ1 on human osteoblastic cells", Head Face Med, 7, pp.13~ (2011)
- Gronthos S, "Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo", Proc Natl Acad Sci USA, 97, pp.13625~13630 (2000)
- Wada Y, "The suppressive effect of enamel matrix derivative on osteocalcin gene expression of osteoblasts is neutralized by an antibody against TGF-ββ", J Periodontol, 79, pp.341~347 (2008)
- Ibbotson KJ, "Effects of transforming growth factors beta 2 on a mouse clonal, osteoblastlike cell line MC3T3-E1", J Bone Miner Res, 4, pp.37~45 (1989)
- Mizuno M, "Carboxyl-terminal propeptide of type I collagen (c-propeptide) modulates the action of TGF-ββ on MC3T3-E1 osteoblastic cells", FEBS Lett, 479, pp.123~126 (2000)
- Neve A, "Ostecalcin: skeletal and extra-skeletal effects", J Cell Physiol, 228, pp.1149~1153 (2013)
- Reseland JE, "The effect of enamel matrix derivative on gene expression in osteoblasts", Eur J Oral Sci, 114(Supplement 1), pp.205~211 (2006)
- Mizuno M, "Bone sialoprotein (BSP) is a crucial factor for the expression of osteoblastic phenotypes of bone marrow cells cultured on type I collagen matrix", Calcif Tissue Int, 66, pp.388~396 (2000)
- Gordon JA, "Bone sialoprotein expression enhances osteoblast differentiation and matrix mineralization in vitro", Bone, 41, pp.462~473 (2007)
- . Wada T, "Calcification of vascular smooth muscle cell cultures: inhibition by osteopontin", Circ Res, 84, pp.166~178 (1999)
- Ishijima M, "Osteopontin is required for mechanical stress-dependent signals to bone marrow cells", J Endocrinol, 193, pp.235~243 (2007)
- Swanson EC, "Amelogenins regulate expression of genes associated with cementoblasts in vitro", Eur J Oral Sci, 114(Supplement 1), pp.239~243 (2006)
- Chen CL, "Comparison of calcium and silicate cement and mineral trioxide aggregate biologic effects and bone markers expression in MG63 cells", J Endod, 35, pp.682~685 (2009)
- Weishaupt P, "Stimulation of osteoblasts with Emdogain increases the expression of specific mineralization markers", Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 106, pp.304~308 (2008)
- Fujisawa R, "Changes in levels of osteonectin in bovine dentine during tooth development", Arch Oral Biol, 34, pp.89~92 (1989)
- Papagerakis P, "Investigation of osteocalcin, osteonectin, and dentin sialophosphoprotein in developing human teeth", Bone, 30, pp.377~385 (2002)
- Sahng G. Kim, "Enamel matrix derivative for replanted teeth in animal models: a systematic review and meta-analysis", Restorative Dentistry and Endodontics, 38(4), pp.194~203 (2013), 대한치과보존학회
