Basic Research| Volume 36, ISSUE 2, P260-263, February 2010

Effect of MTAD on the Differentiation of Osteoblast-like Cells



      The aim of the present study was to investigate the effect of MTAD on the differentiation of osteoblast-like cells.


      The cell viability assay was performed to evaluate the cytotoxicity of MTAD on MC3T3-E1 and periodontal ligament cells at the various concentrations. The bone sialoprotein (BSP) gene expression was also examined by real-time polymerase chain reaction.


      MTAD exhibited a lower cytotoxicity compared with other intracanal irrigants and medication. The MC3T3-E1 cells treated with H2O2 showed a decrease in the alkaline phosphatase (ALP) activity by 40% on day 14 compared with the control group at the concentration of 50 μg/mlL. No significant difference in the ALP activity was observed between MTAD and control group. Furthermore, MTAD and Ca(OH)2 paste did not change in BSP gene expression in MC3T3-E1 cells on day 21.


      These results suggested that MTAD is a less cytotoxic irrigant and does not affect differentiation into osteoblasts compared with other intracanal irrigants, such as H2O2, NaOCl, ethylenediaminetetraacetic acid, and chlorhexidine.

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        • Moller A.J.
        • Fabricius L.
        • Dahlen G.
        • Ohman A.E.
        • Heyden G.
        Influence on periapical tissues of indigenous oral bacteria and necrotic pulp tissue in monkeys.
        Scand J Dent Res. 1981; 89: 475-484
        • Kiryu T.
        • Hoshino E.
        • Iwaku M.
        Bacteria invading periapical cementum.
        J Endod. 1994; 20: 169-172
        • Sen B.H.
        • Piskin B.
        • Demirci T.
        Observation of bacteria and fungi in infected root canals and dentinal tubules by SEM.
        Endod Dent Traumatol. 1995; 11: 6-9
        • Jeansonne M.J.
        • White R.R.
        A comparison of 2.0% chlorhexidine gluconate and 5.25% sodium hypochlorite as antimicrobial endodontic irrigants.
        J Endod. 1994; 20: 276-278
        • Bystrom A.
        • Sundqvist G.
        Bacteriologic evaluation of the efficacy of mechanical root canal instrumentation in endodontic therapy.
        Scand J Dent Res. 1981; 89: 321-328
        • Vertucci F.J.
        Root canal anatomy of the human permanent teeth.
        Oral Surg Oral Med Oral Pathol. 1984; 58: 589-599
        • Torabinejad M.
        • Handysides R.
        • Khademi A.A.
        • Bakland L.K.
        Clinical implications of the smear layer in endodontics: a review.
        Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2002; 94: 658-666
        • Sjogren U.
        • Figdor D.
        • Persson S.
        • Sundqvist G.
        Influence of infection at the time of root filling on the outcome of endodontic treatment of teeth with apical periodontitis.
        Int Endod J. 1997; 30: 297-306
        • Pataky L.
        • Ivanyi I.
        • Grigar A.
        • Fazekas A.
        Antimicrobial efficacy of various root canal preparation techniques: an in vitro comparative study.
        J Endod. 2002; 28: 603-605
        • Zehnder M.
        Root canal irrigants.
        J Endod. 2006; 32: 389-398
        • Cwikla S.J.
        • Belanger M.
        • Giguere S.
        • Progulske-Fox A.
        • Vertucci F.J.
        Dentinal tubule disinfection using three calcium hydroxide formulations.
        J Endod. 2005; 31: 50-52
        • Torabinejad M.
        • Khademi A.A.
        • Babagoli J.
        • et al.
        A new solution for the removal of the smear layer.
        J Endod. 2003; 29: 170-175
        • Torabinejad M.
        • Shabahang S.
        • Aprecio R.M.
        • Kettering J.D.
        The antimicrobial effect of MTAD: an in vitro investigation.
        J Endod. 2003; 29: 400-403
        • Shabahang S.
        • Pouresmail M.
        • Torabinejad M.
        In vitro antimicrobial efficacy of MTAD and sodium hypochlorite.
        J Endod. 2003; 29: 450-452
        • Shabahang S.
        • Torabinejad M.
        Effect of MTAD on Enterococcus faecalis-contaminated root canals of extracted human teeth.
        J Endod. 2003; 29: 576-579
        • Torabinejad M.
        • Cho Y.
        • Khademi A.A.
        • Bakland L.K.
        • Shabahang S.
        The effect of various concentrations of sodium hypochlorite on the ability of MTAD to remove the smear layer.
        J Endod. 2003; 29: 233-239
        • Krause T.A.
        • Liewehr F.R.
        • Hahn C.L.
        The antimicrobial effect of MTAD, sodium hypochlorite, doxycycline, and citric acid on Enterococcus faecalis.
        J Endod. 2007; 33: 28-30
        • Davis J.M.
        • Maki J.
        • Bahcall J.K.
        An in vitro comparison of the antimicrobial effects of various endodontic medicaments on Enterococcus faecalis.
        J Endod. 2007; 33: 567-569
        • Zhang W.
        • Torabinejad M.
        • Li Y.
        Evaluation of cytotoxicity of MTAD using the MTT-tetrazolium method.
        J Endod. 2003; 29: 654-657
        • Fujii S.
        • Maeda H.
        • Wada N.
        • Kano Y.
        • Akamine A.
        Establishing and characterizing human periodontal ligament fibroblasts immortalized by SV40T-antigen and hTERT gene transfer.
        Cell Tissue Res. 2006; 324: 117-125
        • Masillamoni C.R.
        • Kettering J.D.
        • Torabinejad M.
        The biocompatibility of some root canal medicaments and irrigants.
        Int Endod J. 1981; 14: 115-120
        • Mjor I.A.
        Biologic assessment of restorative dental materials: interrelationship of biologic and technologic properties.
        Oper Dent. 1978; 3: 9-13
        • Denizot F.
        • Lang R.
        Rapid colorimetric assay for cell growth and survival: modifications to the tetrazolium dye procedure giving improved sensitivity and reliability.
        J Immunol Methods. 1986; 89: 271-277
        • Ogata K.
        • Imazato S.
        • Ehara A.
        • et al.
        Comparison of osteoblast responses to hydroxyapatite and hydroxyapatite/soluble calcium phosphate composites.
        J Biomed Mater Res. 2005; 72: 127-135
        • Imazato S.
        • Horikawa D.
        • Ogata K.
        • Kinomoto Y.
        • Ebisu S.
        Responses of MC3T3-E1 cells to three dental resin-based restorative materials.
        J Biomed Mater Res. 2006; 76: 765-772
        • Beck Jr., G.R.
        • Sullivan E.C.
        • Moran E.
        • Zerler B.
        Relationship between alkaline phosphatase levels, osteopontin expression, and mineralization in differentiating MC3T3-E1 osteoblasts.
        J Cell Biochem. 1998; 68: 269-280
        • Jadlowiec J.
        • Koch H.
        • Zhang X.
        • Campbell P.G.
        • Seyedain M.
        • Sfeir C.
        Phosphophoryn regulates the gene expression and differentiation of NIH3T3, MC3T3-E1, and human mesenchymal stem cells via the integrin/MAPK signaling pathway.
        J Biol Chem. 2004; 279: 53323-53330
        • Marom R.
        • Shur I.
        • Solomon R.
        • Benayahu D.
        Characterization of adhesion and differentiation markers of osteogenic marrow stromal cells.
        J Cell Physiol. 2005; 202: 41-48
        • Aubin J.E.
        • Liu F.
        • Malaval L.
        • Gupta A.K.
        Osteoblast and chondroblast differentiation.
        Bone. 1995; 17: 77S-83S
        • Malaval L.
        • Wade-Gueye N.M.
        • Boudiffa M.
        • et al.
        Bone sialoprotein plays a functional role in bone formation and osteoclastogenesis.
        J Exp Med. 2008; 205: 1145-1153
        • Leonardo M.R.
        • da Silva L.A.
        • Leonardo Rde T.
        • Utrilla L.S.
        • Assed S.
        Histological evaluation of therapy using a calcium hydroxide dressing for teeth with incompletely formed apices and periapical lesions.
        J Endod. 1993; 19: 348-352