Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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Advances in Clinical and Experimental Medicine

2018, vol. 27, nr 12, December, p. 1615–1623

doi: 10.17219/acem/75776

Publication type: original article

Language: English

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Simvastatin increases cell viability and suppresses the expression of cytokines and vascular endothelial growth factor in inflamed human dental pulp stem cells in vitro

Dai Xue1,A,B,C,D,E,F, Zhongjian Gong2,B,C,D,F, Fangyong Zhu2,C,D,F, Yanjing Qiu3,B,C,E,F, Xiaodan Li1,C,F

1 Department of Stomatology, Wuxi Children's Hospital, Nanjing Medical University, China

2 Department of Stomatology, Wuxi People's Hospital, Nanjing Medical University, China

3 Department of Stomatology, Pudong New Area People's Hospital, Shanghai, China

Abstract

Background. In recent years, simvastatin has been demonstrated to be capable of inducing odontogenic differentiation in human dental pulp stem cells (DPSCs), which makes it a promising source for endodontic treatment in pulpitis. However, a comprehensive understanding of how simvastatin affects the behavior of DPSCs and its potential in pulpitis is still lacking.
Objectives. In this study, we investigated the effects of simvastatin on the viability of inflamed DPSCs. The expression of cytokines and vascular endothelial growth factor (VEGF) was also studied in response to simvastatin treatment.
Material and Methods. We characterized the cell viability, inflammatory reactions and the production of VEGF in inflamed DPSCs, induced by lipopolysaccharides (LPS). The methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, cell cycle, apoptosis analysis, quantitative reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and western blot analyses were performed.
Results. We observed that a low dosage of simvastatin accelerated cell proliferation , whereas its high dosage (>15 μg/mL) suppressed propagation. A simvastatin dose of 8 μg/mL was sufficient to promote cell growth and cell cycle progression in DPSCs treated with LPS. Meanwhile, simvastatin induced apoptosis. The expression of multiple cytokines, including interleukins (IL)-1, IL-4 and IL-1β, and especially interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α), was significantly suppressed. Moreover, the protein secretion and mRNA transcription of VEGF was observed to be markedly inhibited by simvastatin by inactivating mitogen-activated protein kinase (MAPK) signaling.
Conclusion. Taken together, these results suggested that simvastatin might be a potent ingredient to enhance cell proliferation, alleviate inflammation response and attune vasculogenesis in pulpitis.

Key words

cytokines, vascular endothelial growth factor, simvastatin, dental pulp stem cell, pulpitis

References (37)

  1. Michaelson PL, Holland GR. Is pulpitis painful? Int Endod J. 2002;35(10):829–832.
  2. Ahlquist ML, Franzen OG. Inflammation and dental pain in man. Endod Dent Traumatol. 1994;10(5):201–209.
  3. Hahn CL, Liewehr FR. Relationships between caries bacteria, host responses, and clinical signs and symptoms of pulpitis. J Endod. 2007;33(3):213–219.
  4. Khabbaz MG, Anastasiadis PL, Sykaras SN. Determination of endotoxins in caries: Association with pulpal pain. Int Endod J. 2000;33(2):132–137.
  5. Jacinto RC, Gomes BP, Shah HN, Ferraz CC, Zaia AA, Souza-Filho FJ. Quantification of endotoxins in necrotic root canals from symptomatic and asymptomatic teeth. J Med Microbiol. 2005;54(Pt 8):777–783.
  6. Liu H, Gronthos S, Shi S. Dental pulp stem cells. Methods Enzymol. 2006;419:99–113.
  7. Tatullo M, Marrelli M, Shakesheff KM, White LJ. Dental pulp stem cells: Function, isolation and applications in regenerative medicine. J Tissue Eng Regen Med. 2015;9(11):1205–1216.
  8. Na S, Zhang H, Huang F, et al. Regeneration of dental pulp/dentine complex with a three-dimensional and scaffold-free stem-cell sheet-derived pellet. J Tissue Eng Regen Med. 2016;10(3):261–270.
  9. Paino F, Ricci G, De Rosa A, et al. Ecto-mesenchymal stem cells from dental pulp are committed to differentiate into active melanocytes. Eur Cell Mater. 2010;20:295–305.
  10. Ustiashvili M, Kordzaia D, Mamaladze M, Jangavadze M, Sanodze L. Investigation of functional activity human dental pulp stem cells at acute and chronic pulpitis. Georgian Med News.2014;234:19–24.
  11. Fedorowicz Z, van Zuuren EJ, Farman AG, Agnihotry A, Al-Langawi JH. Antibiotic use for irreversible pulpitis. Cochrane Database Syst Rev. 2013;2:CD004969. doi: 10.1002/14651858.CD004969.pub4
  12. Hoskin E, Veitz-Keenan A. Antibiotics are not useful to reduce pain associated with irreversible pulpitis. Evid Based Dent. 2016;17(3):81–82.
  13. Iohara K, Nakashima M, Ito M, Ishikawa M, Nakasima A, Akamine A. Dentin regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 2. J Dent Res. 2004;83(8):590–595.
  14. Saito T, Ogawa M, Hata Y, Bessho K. Acceleration effect of human recombinant bone morphogenetic protein-2 on differentiation of human pulp cells into odontoblasts. J Endod. 2004;30(4):205–208.
  15. Karanxha L, Park SJ, Son WJ, Nor JE, Min KS. Combined effects of simvastatin and enamel matrix derivative on odontoblastic differentiation of human dental pulp cells. J Endod. 2013;39(1):76–82.
  16. Okamoto Y, Sonoyama W, Ono M, et al. Simvastatin induces the odontogenic differentiation of human dental pulp stem cells in vitro and in vivo. J Endod. 2009;35(3):367–372.
  17. Jia W, Zhao Y, Yang J, et al. Simvastatin promotes dental pulp stem cell-induced coronal pulp regeneration in pulpotomized teeth. J Endod. 2016;42(7):1049–1054.
  18. Raoof M, Yaghoobi MM, Derakhshani A, et al. A modified efficient method for dental pulp stem cell isolation. Dent Res J (Isfahan). 2014;11(2):244–250.
  19. Shi X, Gipp J, Dries M, Bushman W. Prostate progenitor cells proliferate in response to castration. Stem Cell Res. 2014;13(1):154–163.
  20. Lee SY, Huang GW, Shiung JN, et al. Magnetic cryopreservation for dental pulp stem cells. Cells Tissues Organs. 2012;196(1):23–33.
  21. Warfvinge J, Dahlen G, Bergenholtz G. Dental pulp response to bacterial cell wall material. J Dent Res. 1985;64(8):1046–1050.
  22. Liu Y, Gao Y, Zhan X, et al. TLR4 activation by lipopolysaccharide and Streptococcus mutans induces differential regulation of proliferation and migration in human dental pulp stem cells. J Endod. 2014;40(9):1375–1381.
  23. Hozhabri NS, Benson MD, Vu MD, et al. Decreasing NF-kappaB expression enhances odontoblastic differentiation and collagen expression in dental pulp stem cells exposed to inflammatory cytokines. PLoS One. 2015;10(1):e0113334. doi: 10.1371/journal.pone.0113334
  24. Guven G, Altun C, Gunhan O, et al. Co-expression of cyclooxygenase-2 and vascular endothelial growth factor in inflamed human pulp: An immunohistochemical study. J Endod. 2007;33(1):18–20.
  25. Dvorak HF, Nagy JA, Feng D, Brown LF, Dvorak AM. Vascular permeability factor/vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. Curr Top Microbiol Immunol. 1999;237:97–132.
  26. Botero TM, Son JS, Vodopyanov D, Hasegawa M, Shelburne CE, Nör JE. MAPK signaling is required for LPS-induced VEGF in pulp stem cells. J Dent Res. 2010;89(3):264–269.
  27. Potdar PD, Jethmalani YD. Human dental pulp stem cells: Applications in future regenerative medicine. World J Stem Cells. 2015;7(5):839–851.
  28. Batouli S, Miura M, Brahim J, et al. Comparison of stem-cell-mediated osteogenesis and dentinogenesis. J Dent Res. 2003;82(12):976–981.
  29. Wada N, Menicanin D, Shi S, Bartold PM, Gronthos S. Immunomodulatory properties of human periodontal ligament stem cells. J Cell Physiol. 2009;219(3):667–676.
  30. Gronthos S, Mankani M, Brahim J, Robey PG, Shi S. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A. 2000;97(25):13625–13630.
  31. Afzali M, Vatankhah M, Ostad SN. Investigation of simvastatin-induced apoptosis and cell cycle arrest in cancer stem cells of MCF 7. J Cancer Res Ther. 2016;12(2):725–730.
  32. Wang G, Cao R, Wang Y, et al. Simvastatin induces cell cycle arrest and inhibits proliferation of bladder cancer cells via PPARγ signalling pathway. Sci Rep. 2016;6:35783. doi: 10.1038/srep35783
  33. Le Blanc K. Mesenchymal stromal cells: Tissue repair and immune modulation. Cytotherapy. 2006;8(6):559–561.
  34. Sonoda S, Yamaza H, Ma L, et al. Interferon-gamma improves impaired dentinogenic and immunosuppressive functions of irreversible pulpitis-derived human dental pulp stem cells. Sci Rep. 2016;6:19286. doi: 10.1038/srep19286
  35. Qin Z, Li Y, Li Y, Liu G. Tumor necrosis factor alpha stimulates proliferation of dental pulp stem cells via Akt/glycogen synthase kinase-3beta/cyclin D1 signaling pathway. J Endod. 2015;41(7):1066–1072.
  36. Chu SC, Tsai CH, Yang SF, et al. Induction of vascular endothelial growth factor gene expression by proinflammatory cytokines in human pulp and gingival fibroblasts. J Endod. 2004;30(10):704–707.
  37. Matsushita K, Motani R, Sakuta T, et al. The role of vascular endothelial growth factor in human dental pulp cells: Induction of chemotaxis, proliferation, and differentiation and activation of the AP-1-dependent signaling pathway. J Dent Res. 2000;79(8):1596–1603.
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