Gene expression profiling in human dental pulp cells treated with mineral trioxide aggregate

Yong-Beom Kim; Won-Jun Shon; WooCheol Lee; Kee-Yeon Kum; Seung-Ho Baek; Kwang-Shik Bae

doi:10.5395/JKACD.2010.35.3.152

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Basic Research Gene expression profiling in human dental pulp cells treated with mineral trioxide aggregate: Yong-Beom Kim, Won-Jun Shon, WooCheol Lee, Kee-Yeon Kum, Seung-Ho Baek, Kwang-Shik Bae; Journal of Korean Academy of Conservative Dentistry 2010;35(3):152-163.
DOI: https://doi.org/10.5395/JKACD.2010.35.3.152
Published online: May 31, 2010

Department of Conservative Dentistry, Dental Research Institute, BK21 Program, School of Dentistry, Seoul National University, Seoul, Korea.

Corresponding Author: Kwang-Shik Bae. Department of Conservative Dentistry, School of Dentistry, Seoul National University, 275-1 Yeongeon-Dong, Jongno-Gu, Seoul, 110-768, Korea. Tel: 82-2-2072-2651, Fax: 82-2-2072-3859, baeks@snu.ac.kr

• Received: March 20, 2010 • Revised: April 6, 2010 • Accepted: April 12, 2010

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Abstract
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Abstract

This study investigated the changes in gene expression when mineral trioxide aggregate (MTA) was applied in vitro to human dental pulp cells (HDPCs). MTA in a teflon tube (diameter 10 mm, height 2 mm) was applied to HDPCs. Empty tube-applied HDPCs were used as negative control. For microarray analysis, total RNA was extracted at 6, 24, and 72 hrs after MTA application. The results were confirmed selectively by performing reverse transcriptase polymerase chain reaction for genes that showed changes of more than two-fold or less than half. Of the 24,546 genes, 109 genes were up-regulated greater than two-fold (e.g., FOSB, THBS1, BHLHB2, EDN1, IL11, FN1, COL10A1, and TUFT1) and 69 genes were down-regulated below 50% (e.g., SMAD6 and DCN). These results suggest that MTA, rather than being a bio-inert material, may have potential to affect the proliferation and differentiation of pulp cells in various ways.
Keywords: Microarray; Mineral trioxide aggregate; Dental pulp capping; Human dental pulp cell; Differentiation; Proliferation

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Figure 1

Differentially expressed genes (178 genes) up- or down-regulated by at least two-fold following a cellular treatment with MTA at every time point. (a) The biological processes of up-regulated 109 genes, (b) The biological processes of down-regulated 69 genes.

Figure 2

Optical density of messenger RNA measured by densitometry.

Table 1

Primer sequence list in RT-PCR

Table 2

Biological processes classification of genes in MTA-treated cells. The genes that showed a difference of more than three-fold, and those considered to be related to differentiation and proliferation out of genes that showed a difference of more than two-fold are listed.

Table 3

The expression level of the messenger RNA determined by RT-PCR. RT-PCR was conducted three times. The relative level of gene expression was normalized against GAPDH messenger RNA, and the control was set as 1.0. Optical density values represent the mean.

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Gene expression profiling in human dental pulp cells treated with mineral trioxide aggregate

J Korean Acad Conserv Dent. 2010;35(3):152-163. Published online May 31, 2010

DOI: https://doi.org/10.5395/JKACD.2010.35.3.152

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Figure

Gene expression profiling in human dental pulp cells treated with mineral trioxide aggregate

Figure 1 Differentially expressed genes (178 genes) up- or down-regulated by at least two-fold following a cellular treatment with MTA at every time point. (a) The biological processes of up-regulated 109 genes, (b) The biological processes of down-regulated 69 genes.

Figure 2 Optical density of messenger RNA measured by densitometry.

Figure 1

Figure 2

Gene expression profiling in human dental pulp cells treated with mineral trioxide aggregate

Table 1

Primer sequence list in RT-PCR

Table 2

Table 3

Table 1 Primer sequence list in RT-PCR

Table 2 Biological processes classification of genes in MTA-treated cells. The genes that showed a difference of more than three-fold, and those considered to be related to differentiation and proliferation out of genes that showed a difference of more than two-fold are listed.

Table 3 The expression level of the messenger RNA determined by RT-PCR. RT-PCR was conducted three times. The relative level of gene expression was normalized against GAPDH messenger RNA, and the control was set as 1.0. Optical density values represent the mean.