Stimulatory Effect of N-acetylcysteine on Odontoblastic Differentiation

  • Jun, Ji-Hae (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University) ;
  • Lee, Hye-Lim (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University) ;
  • Baek, Jeong-Hwa (Department of Cell & Developmental Biology, School of Dentistry and Dental Research Institute, Seoul National University)
  • Published : 2008.12.31

Abstract

Reparative dentine formation requires newly differentiated odontoblast-like cells. Therefore, identification of the molecule that stimulates the odontogenic differentiation of precursor cells in the tooth pulp will be helpful for the development of strategies to repair damaged pulp. In this study, we examined the effect of N-acetylcysteine (NAC) on the odontogenic differentiation of MDPC-23 cells, a mouse odontoblast-like cell line derived from dental papilla, and primary cultured rat dental papilla cells (RDPCs). NAC (1-30 mM) suppressed production of reactive oxygen species in MDPC-23 cells in a dose-dependent manner. Although 5 to 20 mM NAC did not alter MDPC-23 cell proliferation, 1 or 30 mM NAC significantly inhibited it. NAC enhanced mineralized nodule formation and the expression of several odontoblast differentiation-associated genes in both RDPCs and MDPC-23. This NAC stimulatory effect was significant, even at concentrations lower than 1 mM. However, NAC did not stimulate expression of bone morphogenetic protein-2, -4, or -7, which are known to enhance odontogenic differentiation. Since reactive oxygen species are also involved in the pulp toxicity of resin-based restorative materials, these results suggest that NAC may be a promising candidate for supplementation of dental restorative materials in order to enhance reparative dentine formation.

Keywords

References

  1. Aikawa F, Nakatsuka M, Kumabe S, Jue SS, Hayashi H, Shin JW, Iwai Y. Expression of DSPP mRNA during differentiation of human dental pulp-derived cells (HDPC) and transplantation of HDPC using alginate scaffold. Int J Oral Biol. 2006;31:73-9
  2. Aruoma OI, Halliwell B, Hoey BM, Butler J. The antioxidant action of N-acetylcysteine: Its reaction with hydrogen peroxide, hydroxyl radical, superoxide, and hypochlorous acid. Free Radic Biol Med. 1989;6:593-7 https://doi.org/10.1016/0891-5849(89)90066-X
  3. Chen S, Gluhak-Heinrich J, Martinez M, Li T, Wu Y, Chuang HH, Chen L, Dong J, Gay I, MacDougall M. Bone morphogenetic protein 2 mediates dentin sialophosphoprotein expression and odontoblast differentiation via NF-Y signaling. J Biol Chem. 2008;283:19359-70 https://doi.org/10.1074/jbc.M709492200
  4. Cox CF, Subay RK, Ostro E, Suzuki S, Suzuki SH. Tunnel defects in dentin bridges: their formation following direct pulp capping. Oper Dent. 1996;21:4-11
  5. Decup F, Six N, Palmier B, Buch D, Lasfargues JJ, Salih E, Goldberg M. Bone sialoprotein-induced reparative dentinogenesis in the pulp of rat's molar. Clin Oral Investig. 2000;4:110-9 https://doi.org/10.1007/s007840050126
  6. Goldberg M, Six N, Decup F, Buch D, Soheili Majd E, Lasfargues JJ, Salih E, Stanislawski L. Application of bioactive molecules in pulp-capping situations. Adv Dent Res. 2001;15:91-5 https://doi.org/10.1177/08959374010150012401
  7. Goldberg M, Lacerda-Pinheiro S, Jegat N, Six N, Septier D, Priam F, Bonnefoix M, Tompkins K, Chardin H, Denbesten P, Veis A, Poliard A. The impact of bioactive molecules to stimulate tooth repair and regeneration as part of restorative dentistry. Dent Clin North Am. 2006;50:277-98 https://doi.org/10.1016/j.cden.2005.11.008
  8. He H, Yu J, Liu Y, Lu S, Liu H, Shi J, Jin Y. Effects of FGF2 and TGFbeta1 on the differentiation of human dental pulp stem cells in vitro. Cell Biol Int. 2008;32:827-34 https://doi.org/10.1016/j.cellbi.2008.03.013
  9. Higashi T, Okamoto H. Characteristics and effects of calcified degenerative zones on the formation of hard tissue barriers in amputated canine dental pulp. J Endod. 1996;22:168-72 https://doi.org/10.1016/S0099-2399(96)80094-X
  10. Jepsen S, Albers HK, Fleiner B, Tucker M, Rueger D. Recombinant human osteogenic protein-1 induces dentin formation: an experimental study in miniature swine. J Endod. 1997;23:378-82 https://doi.org/10.1016/S0099-2399(97)80187-2
  11. Jun JH, Lee SH, Kwak HB, Lee ZH, Seo SB, Woo KM, Ryoo HM, Kim GS, Baek JH. N-acetylcysteine stimulates osteoblastic differentiation of mouse calvarial cells. J Cell Biochem. 2008;103:1246-55 https://doi.org/10.1002/jcb.21508
  12. Kim do Y, Jun JH, Lee HL, Woo KM, Ryoo HM, Kim GS, Baek JH, Han SB. N-acetylcysteine prevents LPS-induced pro-inflammatory cytokines and MMP2 production in gingival fibroblasts. Arch Pharm Res. 2007;30:1283-92 https://doi.org/10.1007/BF02980269
  13. Kim GH, Song DK, Cho CH, Yoo SK, Kim DK, Park GY, Suh SI, Jang BC, Lim JG. Muscular cell proliferative and protective effects of N-acetylcysteine by modulating activity of extracellular signal-regulated protein kinase. Life Sci. 2006;79:622-8 https://doi.org/10.1016/j.lfs.2006.02.008
  14. Kojima N, Yamada M, Paranjpe A, Tsukimura N, Kubo K, Jewett A, Ogawa T. Restored viability and function of dental pulp cells on poly-methylmethacrylate (PMMA)-based dental resin supplemented with N-acetyl cysteine (NAC). Dent Mater. 2008;24:1686-93 https://doi.org/10.1016/j.dental.2008.04.008
  15. Nakamura Y, Slaby I, Matsumoto K, Ritchie HH, Lyngstadaas SP. Immunohistochemical characterization of rapid dentin formation induced by enamel matrix derivative. Calcif Tissue Int. 2004;75:243-52 https://doi.org/10.1007/s00223-003-0153-y
  16. Nakamura Y, Slaby I, Spahr A, Pezeshki G, Matsumoto K, Lyngstadaas SP. Ameloblastin fusion protein enhances pulpal healing and dentin formation in porcine teeth. Calcif Tissue Int. 2006;78:278-84 https://doi.org/10.1007/s00223-005-0144-2
  17. Nakashima M. Induction of dentin formation on canine amputated pulp by recombinant human bone morphogenetic proteins (BMP)-2 and -4. J Dent Res. 1994a;73:1515-22 https://doi.org/10.1177/00220345940730090601
  18. Nakashima M. Induction of dentine in amputated pulp of dogs by recombinant human bone morphogenetic proteins-2 and -4 with collagen matrix. Arch Oral Biol. 1994b;39:1085-9 https://doi.org/10.1016/0003-9969(94)90062-0
  19. Narayanan K, Gajjeraman S, Ramachandran A, Hao J, George A. Dentin matrix protein 1 regulates dentin sialophosphoprotein gene transcription during early odontoblast differentiation. J Biol Chem. 2006;281:19064-71 https://doi.org/10.1074/jbc.M600714200
  20. Nargi JL, Ratan RR, Griffin DE. p53-independent inhibition of proliferation and p21(WAF1/Cip1)-modulated induction of cell death by the antioxidants N-acetylcysteine and vitamin E. Neoplasia. 1999;1:544-56 https://doi.org/10.1038/sj.neo.7900068
  21. Paranjpe A, Cacalano NA, Hume WR, Jewett A. Nacetylcysteine protects dental pulp stromal cells from HEMA-induced apoptosis by inducing differentiation of the cells. Free Radic Biol Med. 2007;43:1394-408 https://doi.org/10.1016/j.freeradbiomed.2007.07.011
  22. Paranjpe A, Sung EC, Cacalano NA, Hume WR, Jewett A. Nacetyl cysteine protects pulp cells from resin toxins in vivo. J Dent Res. 2008;87:537-41 https://doi.org/10.1177/154405910808700603
  23. Parasassi T, Brunelli R, Bracci-Laudiero L, Greco G, Gustafsson AC, Krasnowska EK, Lundeberg J, Lundeberg T, Pittaluga E, Romano MC, Serafino A. Differentiation of normal and cancer cells induced by sulfhydryl reduction: biochemical and molecular mechanisms. Cell Death Differ. 2005;12:1285-96 https://doi.org/10.1038/sj.cdd.4401663
  24. Ruch JV, Lesot H, Begue-Kirn C. Odontoblast differentiation. Int J Dev Biol. 1995;39:51-68
  25. Rutherford RB, Wahle J, Tucker M, Rueger D, Charette M. Induction of reparative dentine formation in monkeys by recombinant human osteogenic protein-1. Arch Oral Biol. 1993;38:571-6 https://doi.org/10.1016/0003-9969(93)90121-2
  26. Shin MS, Yeon KY, Oh SB, Kim JS. Expression of TRP channels in mouse dental papilla cell-23 (MDPC-23) cell line. Int J Oral Biol. 2006;31:135-40
  27. Smith AJ, Cassidy N, Perry H, Begue-Kirn C, Ruch JV, Lesot H. Reactionary dentinogenesis. Int J Dev Biol. 1995;39:273-80
  28. Stanislawski L, Daniau X, Lauti A, Goldberg M. Factors responsible for pulp cell cytotoxicity induced by resinmodified glass ionomer cements. J Biomed Mater Res. 1999;48:277-88 https://doi.org/10.1002/(SICI)1097-4636(1999)48:3<277::AID-JBM11>3.0.CO;2-T
  29. Stanislawski L, Soheili-Majd E, Perianin A, Goldberg M. Dental restorative biomaterials induce glutathione depletion in cultured human gingival fibroblast: protective effect of Nacetyl cysteine. J Biomed Mater Res. 2000;51:469-74 https://doi.org/10.1002/1097-4636(20000905)51:3<469::AID-JBM22>3.0.CO;2-B
  30. Stanislawski L, Lefeuvre M, Bourd K, Soheili-Majd E, Goldberg M, Perianin A. TEGDMA-induced toxicity in human fibroblasts is associated with early and drastic glutathione depletion with subsequent production of oxygen reactive species. J Biomed Mater Res A. 2003;66:476-82
  31. Sun ZL, Fang DN, Wu XY, Ritchie HH, Begue-Kirn C, Wataha JC, Hanks CT, Butler WT. Expression of dentin sialoprotein (DSP) and other molecular determinants by a new cell line from dental papillae, MDPC-23. Connect Tissue Res. 1998;37:251-61 https://doi.org/10.3109/03008209809002443
  32. Thesleff I. Epithelial-mesenchymal signalling regulating tooth morphogenesis. J Cell Sci. 2003;116:1647-8 https://doi.org/10.1242/jcs.00410
  33. Tziafas D. Basic mechanisms of cytodifferentiation and dentinogenesis during dental pulp repair. Int J Dev Biol. 1995;39:281-90
  34. Tziafas D. The future role of a molecular approach to pulpdentinal regeneration. Caries Res. 2004;38:314-20 https://doi.org/10.1159/000077771
  35. Tziafas D, Alvanou A, Panagiotakopoulos N, Smith AJ, Lesot H, Komnenou A, Ruch JV. Induction of odontoblast-like cell differentiation in dog dental pulps after in vivo implantation of dentine matrix components. Arch Oral Biol. 1995;40:883-93 https://doi.org/10.1016/0003-9969(95)00069-2
  36. Tziafas D, Alvanou A, Papadimitriou S, Gasic J, Komnenou A. Effects of recombinant basic fibroblast growth factor, insulin-like growth factor-II and transforming growth factorbeta 1 on dog dental pulp cells in vivo. Arch Oral Biol. 1998;43:431-44 https://doi.org/10.1016/S0003-9969(98)00026-0
  37. Wu L, Zhu F, Wu Y, Lin Y, Nie X, Jing W, Qiao J, Liu L, Tang W, Zheng X, Tian W. Dentin sialophosphoprotein-promoted mineralization and expression of odontogenic genes in adipose-derived stromal cells. Cells Tissues Organs. 2008;187:103-12 https://doi.org/10.1159/000110079
  38. Ye L, MacDougall M, Zhang S, Xie Y, Zhang J, Li Z, Lu Y, Mishina Y, Feng JQ. Deletion of dentin matrix protein-1 leads to a partial failure of maturation of predentin into dentin, hypomineralization, and expanded cavities of pulp and root canal during postnatal tooth development. J Biol Chem. 2004;279:19141-8 https://doi.org/10.1074/jbc.M400490200