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Calcium Ions Released from Mineral Trioxide Aggregate Convert the Differentiation Pathway of C2C12 Cells into Osteoblast Lineage

2012; Elsevier BV; Volume: 39; Issue: 1 Linguagem: Inglês

10.1016/j.joen.2012.10.006

1878-3554

Sakurako Matsumoto, Makoto Hayashi, Yusuke Suzuki, Naoto Suzuki, Masao Maeno, Bunnai Ogiso,

Oral microbiology and periodontitis research

IntroductionThe purpose of this study was to examine the effect of mineral trioxide aggregate (MTA) on pluripotent-mesenchymal cell differentiation.MethodsThe pluripotent-mesenchymal cell line C2C12 was cultured in a 5% serum medium to induce cell differentiation with or without MTA. The differentiation to myoblasts was analyzed by the immunocytochemical staining of myosin heavy chains. The cellular phenotype-specific markers characterizing the osteoblasts (Runx2 and osterix), chondroblasts (Sox9), myoblasts (MyoD), and adipocytes (LPL) were estimated with mRNA and protein levels by using real-time polymerase chain reaction and Western blot analysis, respectively. To verify that the effect of MTA was caused by the released calcium ions, the mRNA levels were analyzed in the presence or absence of MTA with ethylene glycol tetraacetic acid, calcium chloride, or verapamil.ResultsC2C12 cells cultured without MTA altered their phenotype to myoblasts, exhibiting positive reactions to myosin heavy chains. However, the cells cultured with MTA were strongly inhibited from developing into myoblasts. The mRNA and protein expressions of Runx2, osterix, and Sox9 significantly increased with MTA; the expressions of MyoD and LPL decreased significantly. Calcium chloride addition without MTA presented a significant increase of mRNA levels of Runx2, osterix, and Sox9; ethylene glycol tetraacetic acid addition with MTA presented a significant increase of mRNA levels of MyoD and LPL. Verapamil blocked the stimulating or suppressing effect of MTA on these transcription factors.ConclusionsOur study showed that MTA converted the differentiation pathway of C2C12 cells into osteoblast and/or chondroblast lineages as a result of elution components such as calcium ions from MTA.