Journal of Korean Dental Science

  • 제9권1호
  • /
  • Pages.1-8
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  • 2016
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  • 2005-4742(pISSN)
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  • 2713-7651(eISSN)
대한치의학회 (Korean Academy of Dental Science)
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Evaluation of the Viability of Rat Periodontal Ligament Cells after Storing at 0℃/2 MPa Condition up to One Week: In Vivo MTT Method

  • Jang, Sun Mi (Department of Conservative Dentistry, Yonsei University College of Dentistry) ;
  • Cho, Sin-Yeon (Department of Dentistry, Catholic Kwandong University International St. Mary's Hospital) ;
  • Kim, Eui-Seong (Department of Conservative Dentistry, Yonsei University College of Dentistry) ;
  • Jung, Il-Young (Department of Conservative Dentistry, Yonsei University College of Dentistry) ;
  • Lee, Seung Jong (Department of Conservative Dentistry, Yonsei University College of Dentistry)
  • 투고 : 2016.04.04
  • 심사 : 2016.06.20
  • 발행 : 2016.06.30
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초록

Purpose: The aim of this study was to evaluate the rat periodontal ligament cell viability under $0^{\circ}C/2$ MPa condition up to one week using in vivo 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay. Materials and Methods: As soon as 110 upper molar teeth of rats were extracted, they were stored in Hartman's solution under $0^{\circ}C/2$ MPa condition for 1, 2, 3, 4, and 7 days each. All specimens were treated with in vivo MTT assay and the value of optical density was measured by ELISA reader. These values were statistically analyzed by one-way ANOVA. Result: There was no statistical difference on MTT value between immediate and 1 day storage group. There were statistically significant differences between 1 day and 2 days tsorage, 2 and 3 days storage groups, respectively. Teeth of 34, and 7 days storage groups showed significantly lower MTT valuesc ompared with shorter period storage groups. Conclusion: When the MTT values were substituted in standard curve, 1 day storage group at $0^{\circ}C/2$ MPa condition showed 68% cell viability when compared with immediate group. It dropped to 13% at 2 days, and to less than 5% at 3 days or more.

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참고문헌

  1. Melcher AH. Repair of wounds in the periodontium of the rat. Influence of periodontal ligament on osteogenesis. Arch Oral Biol. 1970; 15: 1183-204. https://doi.org/10.1016/0003-9969(70)90010-5
  2. Schwartz O, Andreasen JO. Cryopreservation of mature teeth before replantation in monkeys (I). Effect of different cryoprotective agents and freezing devices. Int J Oral Surg. 1983; 12: 425-36. https://doi.org/10.1016/S0300-9785(83)80034-9
  3. Schwartz O, Andreasen JO, Greve T. Cryopreservation before replantation of mature teeth in monkeys. (II). Effect of preincubation, different freezing and equilibration rates and endodontic treatment upon periodontal healing. Int J Oral Surg. 1985; 14: 350-61. https://doi.org/10.1016/S0300-9785(85)80025-9
  4. Hupp JG, Mesaros SV, Aukhil I, Trope M. Periodontal ligament vitality and histologic healing of teeth stored for extended periods before transplantation. Endod Dent Traumatol. 1998; 14: 79-83. https://doi.org/10.1111/j.1600-9657.1998.tb00815.x
  5. Soder PO, Otteskog P, Andreasen JO, Modeer T. Effect of drying on viability of periodontal membrane. Scand J Dent Res. 1977; 85: 164-8.
  6. Kim H, Song MJ, Shin SJ, Lee Y, Park JW. Esthetic rehabilitation of single anterior edentulous space using fiber-reinforced composite. Restor Dent Endod. 2014; 39: 220-5. https://doi.org/10.5395/rde.2014.39.3.220
  7. Arav A, Natan Y. Directional freezing: a solution to the methodological challenges to preserve large organs. Semin Reprod Med. 2009; 27: 438-42. https://doi.org/10.1055/s-0029-1241052
  8. Chesne C, Guillouzo A. Cryopreservation of isolated rat hepatocytes: a critical evaluation of freezing and thawing conditions. Cryobiology. 1988; 25: 323-30. https://doi.org/10.1016/0011-2240(88)90040-5
  9. Kim E, Jeon IS, Kim JW, Kim J, Jung HS, Lee SJ. An MTT-based method for quantification of periodontal ligament cell viability. Oral Dis. 2007; 13: 495-9. https://doi.org/10.1111/j.1601-0825.2006.01328.x
  10. Pegg DE. Cryopreservation of vascular endothelial cells as isolated cells and as monolayers. Cryobiology. 2002; 44: 46-53. https://doi.org/10.1016/S0011-2240(02)00003-2
  11. Rubinsky B. Principles of low temperature cell preservation. Heart Fail Rev. 2003; 8: 277-84. https://doi.org/10.1023/A:1024734003814
  12. Lee Y. Evaluation of rat periodontal ligament cell viability after hypothermic preservation under high hydrostatic pressure [PhD dissertation]. Seoul: Graduate School, Yonsei University; 2012.
  13. Hupp JG, Trope M, Mesaros SV, Aukhil I. Tritiated thymidine uptake in periodontal ligament cells of dogs' teeth stored in various media for extended time periods. Endod Dent Traumatol. 1997; 13: 223-7. https://doi.org/10.1111/j.1600-9657.1997.tb00044.x
  14. Narayanan AS, Page RC. Connective tissues of the periodontium: a summary of current work. Coll Relat Res. 1983; 3: 33-64. https://doi.org/10.1016/S0174-173X(83)80048-X
  15. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65: 55-63. https://doi.org/10.1016/0022-1759(83)90303-4
  16. Khedmat S, Dehghan S, Hadjati J, Masoumi F, Nekoofar MH, Dummer PM. In vitro cytotoxicity of four calcium silicate-based endodontic cements on human monocytes, a colorimetric MTT assay. Restor Dent Endod. 2014; 39: 149-54. https://doi.org/10.5395/rde.2014.39.3.149
  17. Henze U, Lennartz A, Hafemann B, Goldmann C, Kirkpatrick CJ, Klosterhalfen B. The influence of the C1-inhibitor BERINERT and the proteinfree haemodialysate ACTIHAEMYL20% on the evolution of the depth of scald burns in a porcine model. Burns. 1997; 23: 473-7. https://doi.org/10.1016/S0305-4179(97)00019-3
  18. Chung JH, Kim J, Choi SH, Kim ES, Park J, Lee SJ. The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure. J Korean Acad Conserv Dent. 2010; 35: 285-94. https://doi.org/10.5395/JKACD.2010.35.4.285
  19. Baek DY, Lee SJ, Jung HS, Kim E. Comparison of viability of oral epithelial cells stored by different freezing methods. J Korean Acad Conserv Dent. 2009; 34: 491-9. https://doi.org/10.5395/JKACD.2009.34.6.491
  20. Du Y, Pribenszky CS, Molnar M, Zhang X, Yang H, Kuwayama M, Pedersen AM, Villemoes K, Bolund L, Vajta G. High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification. Reproduction. 2008; 135: 13-7. https://doi.org/10.1530/REP-07-0362
  21. Pribenszky C, Molnar M, Cseh S, Solti L. Improving post-thaw survival of cryopreserved mouse blastocysts by hydrostatic pressure challenge. Anim Reprod Sci. 2005; 87: 143-50. https://doi.org/10.1016/j.anireprosci.2004.09.007
  22. Jeon IS, Kim ES, Kim J, Lee SJ. Evaluation of periodontal ligament cell viability in rat teeth according to various extra-oral dry storage times using MTT assay. J Korean Acad Conserv Dent. 2006; 31: 398-408. https://doi.org/10.5395/JKACD.2006.31.5.398
  23. Kim JW, Kim ES, Kim J, Lee SJ. Evaluation of periodontal ligament cell viability in rat teeth after frozen preservation using in-vivo MTT assay. J Korean Acad Conserv Dent. 2006; 31: 192-202. https://doi.org/10.5395/JKACD.2006.31.3.192
  24. Amemiya H, Matsuzaka K, Kokubu E, Ohta S, Inoue T. Cellular responses of rat periodontal ligament cells under hypoxia and re-oxygenation conditions in vitro. J Periodontal Res. 2008; 43: 322-7. https://doi.org/10.1111/j.1600-0765.2007.01032.x
  25. Amemiya K, Kaneko Y, Muramatsu T, Shimono M, Inoue T. Pulp cell responses during hypoxia and reoxygenation in vitro. Eur J Oral Sci. 2003; 111: 332-8. https://doi.org/10.1034/j.1600-0722.2003.00047.x
  26. Smaili SS, Hsu YT, Youle RJ, Russell JT. Mitochondria in Ca2+ signaling and apoptosis. J Bioenerg Biomembr. 2000; 32: 35-46. https://doi.org/10.1023/A:1005508311495