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

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

doi:10.1016/j.joen.2012.10.006

Cited by 63 publications

(66 citation statements)

References 33 publications

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“…The number of cultured cells in the presence or absence of MTA was determined by using the Cell Counting Kit 8 (Dojindo Molecular Technologies, Kumamoto, Japan) on days 1, 2, and 3, as described previously 14) . Briefly, the medium was replaced with fresh medium containing 10% (v/v) cell counting reagent at each time point, and the incubation was continued for 2 h. After incubation, the absorbance of the reaction products was measured at 450 nm using a microtiter plate reader (SpectraMax 190, Molecular Devices, Sunnyvale, CA, USA).…”

Section: Determination Of Cell Numbermentioning

confidence: 99%

“…Takita et al reported that the release of ions from MTA provides the optimum amount of calcium for the proliferation and migration of dental pulp cells 4,13) . In another study involving undifferentiated mesenchymal cell cultures, calcium ions from MTA induced differentiation into osteoblast lineages 14) . Given these findings, the release of calcium ions from MTA may be an important characteristic underlying the biological effect of MTA.…”

Section: Introductionmentioning

confidence: 98%

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Involvement of the calcium-sensing receptor in mineral trioxide aggregate-induced osteogenic gene expression in murine MC3T3-E1 cells

Yasukawa

Hayashi

Tanabe

et al. 2017

Dental Materials Journal

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Mineral trioxide aggregate (MTA) has excellent biocompatibility as well as bioactivity, including an ability to induce osteoblast differentiation. We examined the effects of the calcium-sensing receptor (CaSR) on osteogenic gene expression induced by MTA. MC3T3-E1 cells were cultured with or without (control) MTA. The expression levels of Runx2, type I collagen, and CaSR genes were analyzed by real-time polymerase chain reaction and their products were measured using enzyme-linked immunosorbent assays. The levels were increased significantly in cells exposed to MTA compared with control. Next, MC3T3-E1 cells were cultured with MTA and EGTA (a calcium chelator), because calcium ions were released continuously from MTA into the culture. Expression levels were decreased to control levels by MTA plus EGTA. NPS2143 (a CaSR antagonist) also reduced MTA-induced gene expression. These results suggest that MTA induced osteogenic gene expressions of Runx2 and type I collagen via CaSR in MC3T3-E1 cells.

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Section: Determination Of Cell Numbermentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Involvement of the calcium-sensing receptor in mineral trioxide aggregate-induced osteogenic gene expression in murine MC3T3-E1 cells

Yasukawa

Hayashi

Tanabe

et al. 2017

Dental Materials Journal

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“…The levels of calcium ions released from MTA are reported to be optimal for cell proliferation 9) and mineralization 22) . We recently reported that MTA converts the differentiation pathway of C2C12 cells to osteoblast and/or chondroblast lineages through the influx of elution components, such as calcium ions released from MTA 13) . The purpose of this study was to clarify the influx of calcium ions released from MTA into C2C12 cells; thus, we investigated changes in intracellular calcium resulting from the presence of MTA.…”

Section: Discussionmentioning

confidence: 99%

“…The progenitor cells of myoblasts, C2C12 cells, originate from undifferentiated mesenchymal cells and have the capacity to differentiate into osteoblasts, chondroblasts, myoblasts or adipocytes 12) . We previously reported that MTA converts the differentiation pathway of C2C12 cells into osteoblast lineages, possibly through an influx of elution components, such as calcium ions released from MTA 13) . However, the intracellular calcium ion concentration of C2C12 cells treated with MTA has not been determined.…”

Section: Introductionmentioning

confidence: 99%

Calcium Ions Released from Mineral Trioxide Aggregate are Taken Up by C2C12 Cells via the L-Type Voltage-Dependent Calcium Channel

Matsumoto¹,

Hayashi²,

Tanabe³

et al. 2013

J. Hard Tissue Biology.

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Mineral trioxide aggregate (MTA) is a commonly used endodontic repair material reported to exhibit calcified-tissue conductive activity. The effect of MTA on osteoblast differentiation has been well-defined; however, its effect on mesenchymal cells is not fully understood. Recently, we reported that MTA converts the differentiation pathway of pluripotent mesenchymal cells (C2C12 cells) into osteoblast lineages, possibly through the influx of elution components, such as calcium ions released from MTA. The purpose of this study was to investigate whether calcium ions released from MTA flow directly into C2C12 cells and whether this influx increases intracellular calcium levels. C2C12 cells were loaded with fluo-3 acetoxymethyl ester, which acts as a calcium-sensitive fluorescent probe. The intracellular calcium levels of C2C12 cells were assessed by confocal laser scanning microscopy. MTA induced a rapid increase in fluorescence intensity (FI) in C2C12 cells; however, application of Dulbecco's modified Eagle's medium (DMEM) as the growth medium had no such effect. To clarify the specific effects of MTA-derived calcium ions, we used calcium chloride as an exogenous calcium ion source and ethylene glycol tetraacetic acid (EGTA) as a calcium chelator. DMEM containing calcium chloride elicited a rapid rise in FI, whereas MTA plus EGTA did not affect FI. Moreover, MTA did not affect FI in C2C12 cells pre-incubated with verapamil, an L-type voltage-dependent calcium channel (VDCC L ) blocker. Our data suggest that calcium ions released from MTA flow directly into C2C12 cells via VDCC L .

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“…Also, other factors [59,60], and signaling pathways [61] are essential for osteoblast differentiation. These regulators interact with each other as networks to trigger diverse signals and orchestrate the transcription of genes crucial to define osteoblastic lineage and differentiation [62][63][64].…”

Section: Transcriptome Analysismentioning

confidence: 99%

Searching for the Molecular Pathways Regulating Bone Mineral Density in the Proteome and RNA Interference Era

Saad¹

2013

J Orth Rheumatol

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Osteoporosis is a polygenic disorder associated with low bone mineral density and deterioration of bone microarchitecture with increased chance of bone fractures. Although bone matrix mineralization and osteoporosis are closely related, the mineralization of bone matrix is almost a forgotten aspect in osteoporosis research. The complex processes of bone matrix mineralization and bone remodeling are tightly regulated by several transcription factors and signal transduction pathways. However, signal transduction pathways occurring at a protein level that depends not only on mRNA transcriptional regulation but also on a multitude of translational and posttranslational controls. Furthermore, proteomics allow a discerning view of complex molecular pathways, provides an efficient method to determine protein candidates, and elucidates signal transduction pathways that regulate bone mineral density and accelerates the discovery of osteoporosis causative genes. RNA interference is a powerful tool for rapid analysis of gene functions. Therefore, strategies to combine proteomics with RNA interference and transgenic RNAi would greatly improve the efficiency of gene discovery and divulge the molecular pathways involved in osteoporosis pathophysiology. In this review, current methods employed to identify genes involved in osteoporosis, which include linkage analysis, candidate gene association studies, genome wide association studies, transcriptome microarray, and proteomics are evaluated, and a new strategy is proposed.

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

Cited by 63 publications

References 33 publications

Involvement of the calcium-sensing receptor in mineral trioxide aggregate-induced osteogenic gene expression in murine MC3T3-E1 cells

Involvement of the calcium-sensing receptor in mineral trioxide aggregate-induced osteogenic gene expression in murine MC3T3-E1 cells

Calcium Ions Released from Mineral Trioxide Aggregate are Taken Up by C2C12 Cells via the L-Type Voltage-Dependent Calcium Channel

Searching for the Molecular Pathways Regulating Bone Mineral Density in the Proteome and RNA Interference Era

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