Biomaterials Research (생체재료학회지)

Korean Society for Biomaterials (한국생체재료학회)
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Mycoplasma detection and elimination are necessary for the application of stem cell from human dental apical papilla to tissue engineering and regenerative medicine

  • Kim, Byung-Chul (Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University) ;
  • Kim, So Yeon (Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University) ;
  • Kwon, Yong-Dae (Department of Oral Maxillofacial Surgery, School of Dentistry, Kyung Hee University) ;
  • Choe, Sung Chul (Department of Pediatric Dentistry, School of Dentistry, Kyung Hee University) ;
  • Han, Dong-Wook (Department of Cogno-Mechatronics Engineering, Pusan National University) ;
  • Hwang, Yu-Shik (Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University)
  • Received : 2014.12.08
  • Accepted : 2015.03.03
  • Published : 2015.06.30
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Abstract

Background: Recently, postnatal stem cells from dental papilla with neural crest origin have been considered as one of potent stem cell sources in regenerative medicine regarding their multi-differentiation capacity and relatively easy access. However, almost human oral tissues have been reported to be infected by mycoplasma which gives rise to oral cavity in teeth, and mycoplasma contamination of ex-vivo cultured stem cells from such dental tissues and its effect on stem cell culture has received little attention. Results: In this study, mycoplama contamination was evaluated with stem cells from apical papilla which were isolated from human third molar and premolars from various aged patients undergoing orthodontic therapy. The ex-vivo expanded stem cells from apical papilla were found to express stem cell markers such as Stro-1, CD44, nestin and CD133, but mycoplama contamination was detected in almost all cell cultures of the tested 20 samples, which was confirmed by mycoplasma-specific gene expression and fluorescence staining. Such contaminated mycoplasma could be successfully eliminated using elimination kit, and proliferation test showed decreased proliferation activity in mycoplasma-contaminated cells. After elimination of contaminated mycoplasma, stem cells from apical papilla showed osteogenic and neural lineage differentiation under certain culture conditions. Conclusion: Our study proposes that the evaluation of mycoplasma contamination and elimination process might be required in the use of stem cells from apical papilla for their potent applications to tissue engineering and regenerative medicine.

Keywords

Acknowledgement

Supported by : National Research Foundation of Korea (NRF)

References

  1. Chai Y, Jiang X, Ito Y, Bringas P, Han J, Rowitch DH, et al. Fate of the mammalian cranial neural crest during tooth and mandibular morphogenesis. Development. 2000;127:1671-9.
  2. Miletich I, Sharpe PT. Neural crest contribution to mammalian tooth formation. Birth Defects Res (Part C). 2004;72:200-12. https://doi.org/10.1002/bdrc.20012
  3. Ikeda E, Hirose M, Kotobuki N, Shimaoka H, Tadokoro M, Maeda M, et al. Osteogenic differentiation of human dental papilla mesenchymal cells. Biochem Biophy Res Comm. 2006;342:1257-62. https://doi.org/10.1016/j.bbrc.2006.02.101
  4. Sakai VT, Zhang Z, Dong Z, Neiva KG, Machado MAAM, Shi S, et al. SHED differentiate into functional odontoblasts and endothelium. J Dent Res. 2010;89:791-6. https://doi.org/10.1177/0022034510368647
  5. Zhang W, Walboomers XF, Shi S, Fan M, Jansen JA. Multilineage differentiation potential of stem cells derived from human dental pulp after cryopreservation. Tissue Eng. 2006;12:2813-23. https://doi.org/10.1089/ten.2006.12.2813
  6. 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. 2000;97:13625-30. https://doi.org/10.1073/pnas.240309797
  7. Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 2004;364:149-55. https://doi.org/10.1016/S0140-6736(04)16627-0
  8. Morsczeck C, Gotz W, Schierholz J, Zeilhofer F, Kuhn U, Mohl C, et al. Isolation of precursor cells (PCs) from human dental follicle of wisdom teeth. Matrix Biol. 2005;24:155-65. https://doi.org/10.1016/j.matbio.2004.12.004
  9. Kim BC, Bae HJ, Kwon IK, Lee EJ, Park JH, Khademhosseini A, et al. Osteoblastic/cementoblastic and neural differentiation of dental stem cells and their applications to tissue engineering and regenerative medicine. Tissue Eng Part B. 2012;18(3):235-44.
  10. Rothova M, Feng J, Sharpe PT, Peterkova R, Tucker AS. Contribution of mesoderm to the developing dental papilla. Inter J Dev Biol. 2011;55(1):59-64. https://doi.org/10.1387/ijdb.103083mr
  11. Ikeda E, Morita R, Nakao K, Ishida K, Nakamura T, Takano-Yamamoto T, et al. Fully functional bioengineered tooth replacement as an organ replacement therapy. Proc Natl Acad Sci. 2009;106:13475-80. https://doi.org/10.1073/pnas.0902944106
  12. Laino G, d'Aquino R, Graziano A, Lanza V, Carinci F, Naro F, et al. A new population of human adult dental pulp stem cells: a useful source of living autologous fibrous bone tissue (LAB). J Bone Miner Res. 2005;20:1394-402. https://doi.org/10.1359/JBMR.050325
  13. Papaccio G, Graziano A, d'Aquino R, Graziano MF, Pirozzi G, Menditti D, et al. Long-term cryopreservation of dental pulp stem cells (SBP-DPSCs) and their differentiated osteoblasts: a cell source for tissue repair. J Cell Phy. 2006;208:319-25. https://doi.org/10.1002/jcp.20667
  14. Arthur A, Rychkov G, Shi S, Koblar SA, Gronthos S. Adult human dental pulp stem cells differentiate toward functionally active neurons under appropriate environmental cues. Stem Cells. 2008;26:1787-1795. https://doi.org/10.1634/stemcells.2007-0979
  15. Kraus DC, Taylor-Robinson D. Mycoplasmas which infect humans. Mycoplasmas. In: Maniloff J, editor. Molecular biology and pathogenesis. Washington DC: American Society for Microbiology Press; 1992. p. 417-44.
  16. Nicolson GL, Nasralla MY, Nicolson NL. The pathogenesis and treatment of mycoplasmal infections. Antimicro Infect Dis News. 1999;17(11):81-7. https://doi.org/10.1016/S1069-417X(00)88885-8
  17. Mcgarrity GJ, Vanaman V, Sarama J. Cytogenetic effects of mycoplasmal infection of cell cultures: A review. In Vitro. 1984;20(1):1-18. https://doi.org/10.1007/BF02633326
  18. Muhlradt PF, Meyer H, Jansen R. Identification of S-(2,3-Dihydroxypropyl) cystein in a macrophage-activating lipopeptide from Mycoplasma fermentans. Biochemistry. 1996;35:7781-6. https://doi.org/10.1021/bi9602831
  19. Hwang YS, Cho J, Chan KLA, Heng J, Boccacini AR, Kazarian SG, et al. The use of murine embryonic stem cells, alginate encapsulation, and rotary microgravity bioreactor in bone tissue engineering. Biomaterials. 2009;30:499-507. https://doi.org/10.1016/j.biomaterials.2008.07.028
  20. Arthur A, Shi S, Zannettino AC, Fujii N, Gronthos S, Koblar SA. Implanted adult human dental pulp stem cells induce endogenous axon guidance. Stem Cells. 2009;27:2229-37. https://doi.org/10.1002/stem.138
  21. Sonoyama W, Liu Y, Yamaza T, Tuan RS, Wang S, Shi S, et al. Characterization of the apical papilla and its residing stem cells from human immature permanent teeth: a pilot study. J Endodontics. 2008;34:166-71. https://doi.org/10.1016/j.joen.2007.11.021
  22. Wislet-Gendebien S, Hans G, Leprince P, Rigo JM, Moonen G, Rogister B. Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype. Stem Cells. 2005;23:392-402. https://doi.org/10.1634/stemcells.2004-0149
  23. Tucker A, Sharpem P. The cutting-edge of mammalian development; how the embryo makes teeth. Nat Rev. 2004;5:499-508.

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