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

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

Article information

Restor Dent Endod. 2010;35(3):152-163
Publication date (electronic) : 2010 May 31
doi : https://doi.org/10.5395/JKACD.2010.35.3.152
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 2010 March 20; Revised 2010 April 06; Accepted 2010 April 12.

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.

References

1. American Association of Endodontists. Glossary of endodontic terms 2003. 7th edth ed. 40.
2. Cox CF, Subay RK, Ostro E, Suzuki S, Suzuki SH. Tunnel defects in dentin bridges: their formation following direct pulp capping. Oper Dent 1996. 214–11.
3. Cox CF, Hafez AA, Akimoto N, Otsuki M, Suzuki S, Tarim B. Biocompatibility of primer, adhesive and resin composite systems on non-exposed and exposed pulps of non-human primate teeth. Am J Dent 1998. 11s55–s63.
4. Cox CF, Tarim B, Kopel H, Gürel G, Hafez A. Technique sensitivity: biological factors contributing to clinical success with various restorative materials. Adv Dent Res 2001. 1585–90.
5. Torabinejad M, Chivian N. Clinical applications of mineral trioxide aggregate. J Endod 1999. 25197–205.
6. Torabinejad M, Hong CU, Pitt-Ford TR, Kettering JD. Cytotoxicity of four root end filling materials. J Endod 1995. 21489–492.
7. Dominguez MS, Witherspoon DE, Gutmann JL, Opperman LA. Histological and scanning electron microscopy assessment of various vital pulp-therapy materials. J Endod 2003. 29324–333.
8. Ford TR, Torabinejad M, Abedi HR, Bakland LK, Kariyawasam SP. Using mineral trioxide aggregate as a pulp-capping material. J Am Dent Assoc 1996. 1271491–1494.
9. Yun YR, Yang IS, Hwang YC, Hwang IN, Choi HR, Yoon SJ, Kim SH, Oh WM. Pulp response of mineral trioxide aggregate, calcium sulfate or calcium hydroxide. J Korean Acad Conserv Dent 2007. 3295–101.
10. Bae JH, Kim YG, Yoon PY, Cho BH, Choi YH. Histological study. J Korean Acad Conserv Dent 2010. 355–12.
11. Andelin W, Shabahang S, Wright K, Torabinejad M. Identification of hard tissue after experimental pulp capping using dentin sialoprotein (DSP) as a marker. J Endod 2003. 29646–650.
12. Kuratate M, Yoshiba K, Shigetani Y, Yoshiba N, Ohshima H, Okiji T. Immunohistochemical analysis of nestin, osteopontin, and proliferating cells in the reparative process of exposed dental pulp capped with mineral trioxide aggregate. J Endod 2008. 34970–974.
13. Min KS, Yang SH, Kim EC. The Combined Effect of Mineral Trioxide Aggregate and Enamel Matrix Derivative on Odontoblastic Differentiation in Human Dental Pulp Cells. J Endod 2009. 35847–851.
14. Brown PO, Botstein D. Exploring the new world of the genome with DNA microarrays. Nat Genet 1999. 2133–37.
15. Shi S, Robey PG, Gronthos S. Comparison of human dental pulp and bone marrow stromal stem cells by cDNA microarray analysis. Bone 2001. 29532–539.
16. McLachlan JL, Smith AJ, Bujalska IJ, Cooper PR. Gene expression profiling of pulpal tissue reveals the molecular complexity of dental caries. Biochim Biophys Acta 2005. 1741271–281.
17. Syudo M, Yamada S, Yanagiguchi K, Matsunaga T, Hayashi Y. Early gene expression analyzed by a genome microarray and real-time PCR in osteoblasts cultured with a 4-META/MMA-TBB adhesive resin sealer. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009. 107e77–e81.
18. So H, Park SH, Choi GW. The comparison of gene expression from human dental pulp cells and periodontal ligament cells. J Korean Acad Conserv Dent 2009. 34430–441.
19. Yokose S, Kadokura H, Tajima Y. Establishment and characterization of a culture system for enzymatically released rat dental pulp cells. Calcif Tissue Int 2000. 66139–144.
20. Goldberg M, Farges JC, Lacerda-Pinheiro S. Inflammatory and immunological aspects of dental pulp repair. Pharmacol Res 2008. 58137–147.
21. Matsui S, Takeuchi H, Tsujimoto Y, Matsushima K. Effects of Smads and BMPs induced by Ga-Al-As laser irradiation on calcification ability of human dental pulp cells. J Oral Sci 2008. 5075–81.
22. Imamura T, Takase M, Nishihara A, Oeda E, Hanai J, Kawabata M, Miyazono K. Smad6 inhibits signaling by the TGF-β superfamily. Nature 1997. 389622–626.
23. Keklikoglu N. The localization of Fos B, a member of transcription factor AP-1 family, in rat odontoblasts and pulpal undifferentiated ectomesenchymal cells. Folia Histochem Cytobiol 2004. 42191–193.
24. Sabatakos G, Sims NA, Chen J, Aoki K, Kelz MB, Amling M, Bouali Y, Mukhopadhyay K, Ford K, Nestler EJ, Baron R. Overexpression of DeltaFosB transcription factor(s) increases bone formation and inhibits adipogenesis. Nat Med 2000. 6985–990.
25. Shen M, Yoshida E, Yan W, Kawamoto T, Suardita K, Koyano Y, Fujimoto K, Noshiro M, Kato Y. Basic helix-loop-helix protein DEC1 promotes chondrocyte differentiation at the early and terminal stages. J Biol Chem 2002. 27750112–50120.
26. Iwata T, Kawamoto T, Sasabe E, Miyazaki K, Fujimoto K, Noshiro M, Kurihara H, Kato Y. Effects of overexpression of basic helix-loop-helix transcription factor Dec1 on osteogenic and adipogenic differentiation of mesenchymal stem cells. Eur J Cell Biol 2006. 85423–431.
27. Lawler J. The functions of thrombospondin-1 and-2. Curr Opin Cell Biol 2000. 12634–640.
28. Murphy-Ullrich JE, Schultz-Cherry S, Höök M. Transforming growth factor-beta complexes with thrombospondin. Mol Biol Cell 1992. 3181–188.
29. Ueno A, Yamashita K, Nagata T, Tsurumi C, Miwa Y, Kitamura S, Inoue H. cDNA cloning of bovine thrombospondin 1 and its expression in odontoblasts and predentin. Biochim Biophys Acta 1998. 138217–22.
30. Casasco A, Calligaro A, Casasco M, Springall DR, Tenti P, Marchetti C. Immunohistochemical localization of endothelin-like immunoreactivity in human tooth germ and mature dental pulp. Anat Embryol (Berl) 1991. 183515–520.
31. Neuhaus SJ, Byers MR. Endothelin receptors and endothelin-1 in developing rat teeth. Arch Oral Biol 2007. 52655–662.
32. Guidry C, Hook M. Endothelins produced by endothelial cells promote collagen gel contraction by fibroblasts. J Cell Biol 1991. 115873–880.
33. Marini M, Carpi S, Bellini A, Patalano F, Mattoli S. Endothelin-1 induces increased fibronectin expression in human bronchial epithelial cells. Biochem Biophys Res Commun 1996. 220896–899.
34. Khan ZA, Farhangkhoee H, Mahon JL, Bere L, Gonder JR, Chan BM. Endothelins: regulators of extracellular matrix protein production in diabetes. Exp Biol Med (Maywood) 2006. 2311022–1029.
35. Yan Y, Liu Z, Zhang WG. In vitro study of the effects of endothelin-1 on human dental pulp cells. Chin J Dent Res 1999. 25–13.
36. Kido S, Kuriwaka-Kido R, Imamura T, Ito Y, Inoue D, Matsumoto T. Mechanical stress induces Interleukin-11 expression to stimulate osteoblast differentiation. Bone 2009. 451125–1132.
37. Suga K, Saitoh M, Fukushima S, Takahashi K, Nara H, Yasuda S. Interleukin-11 induces osteoblast differentiation and acts synergistically with bone morphogenetic protein-2 in C3H10T1/2 cells. J Interferon Cytokine Res 2001. 21695–707.
38. Takeuchi Y, Watanabe S, Ishii G, Takeda S, Nakayama K, Fukumoto S. Interleukin-11 as a stimulatory factor for bone formation prevents bone loss with advancing age in mice. J Biol Chem 2002. 27749011–49018.
39. Hynes R. Molecular biology of fibronectin. Ann Rev Cell Biol 1985. 167–90.
40. Lesot H, Osman M, Ruch JV. Immunofluorescent localization of collagens, fibronectin and laminin during terminal differentiation of odontoblasts. Dev Biol 1981. 82371–381.
41. Yoshiba N, Yoshiba K, Twaku M, Nakamura H, Osawa H. A confocal laser scanning microscopic study of the immunofluorescent localization of FN in the odontoblast layer of human teeth. Arch Oral Biol 1994. 39395–400.
42. Wang P, Hao J, Shi J. Actions of bovine plasma fibronectin on cultured human dental pulp cells. Chin J Dent Res 2000. 355–62.
43. Hoshi K, Kemmotsu S, Takeuchi Y, Amizuka N, Ozawa H. The primary calcification in bones follows removal of decorin and fusion of collagen fibrils. J Bone Miner Res 1999. 14273–280.
44. Mochida Y, Duarte WR, Tanzawa H, Paschalis EP, Yamauchi M. Decorin modulates matrix mineralization in vitro. Biochem Biophys Res Commun 2003. 3056–9.
45. Alini M, Marriott A, Chen T, Abe S, Poole AR. A novel angiogenic molecule produced at the time of chondrocyte hypertrophy during endochondral bone formation. Dev Biol 1996. 176124–132.
46. Felszeghy S, Hollo K, Modis L, Lammi MJ. Type X collagen in human enamel development: a possible role in mineralization. Acta Odontol Scand 2000. 58171–176.
47. Kwan KM, Pang MK, Zhou S, Cowan SK, Kong RY, Pfordte T. Abnormal compartmentalization of cartilage matrix components in mice lacking collagen X: Implications for function. J Cell Biol 1997. 136459–471.
48. Deutsch D, Palmon A, Fisher LW, Kolodny N, Termine JD, Young MF. Sequencing of bovine enamelin ("tuftelin") a novel acidic enamel protein. J Biol Chem 1991. 26616021–16028.
49. Paine CT, Paine ML, Luo W, Okamoto CT, Lyngstadaas SP, Snead ML. A tuftelin-interacting protein (TIP39) localizes to the apical secretory pole of mouse ameloblasts. J Biol Chem 2000. 27522284–22292.

Article information Continued

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 1

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 2

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.

Table 3