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HOME > Restor Dent Endod > Volume 34(4); 2009 > Article
Original Article THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS
Young-Eun Lee1, Eui-Seong Kim1, Jin Kim2, Seung-Hoon Han3, Seung-Jong Lee1,*
J Korean Acad Conserv Dent 2009;34(4):-363.
DOI: https://doi.org/10.5395/JKACD.2009.34.4.356
Published online: January 14, 2009

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*Corresponding Author: Seung-Jong Lee, Department of Conservative Dentistry, College of Dentistry, Yonsei University, 250 Seongsan-ro Seodaemin-Ku, Seoul, 120-752, Korea, Tel: 82-2-2228-3148 Fax: 82-2-313-7575, E-mail sjlee@yuhs.ac
• Received: April 23, 2009   • Revised: June 16, 2009   • Accepted: June 22, 2009

Copyright © 2009 The Korean Academy of Conservative Dentistry

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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  • The purpose of this study was to evaluate the viability of periodontal ligament cells in rat teeth using slow cryo-preservation method under pressure by means of MTT assay and WST-1 assay. Eighteen teeth of Sprague-Dawley white female rats of 4 week-old were used for each group.
    Both sides of the first and second maxillary molars were extracted as atraumatically as possible under Tiletamine anesthesia. The experimental groups were group 1 (Immediate control), group 2 (Cold preservation at 4°C for 1 week), group 3 (Slow freezing), group 4 (Slow freezing under pressure of 3 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37°C water bath, then MTT assay and WST-1 assay were processed. One way ANOVA and Tukey method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization.
    In both MTT and WST-1 assay, group 4 showed significantly higher viability of periodontal ligament cells than group 2 and 3 (p < 0.05), but showed lower viability than immediate control group.
    By the results of this study, slow cryo-preservation method under pressure suggests the possibility for long term cryo-preservation of the teeth.
  • Keywords: periodontal ligament cell; MTT; WST-1; viability; pressure; cryo-preservation
Figure 1.
Schematic diagram of program freezer with pressure vessel.

  1. Oxygen container : 3MPa of pressure

  2. Program freezer

  3. Pressure bottle

  4. 2ml Cryotube: 1ml F medium + 10% DMSO

  5. Tooth treated 10% DMSO

  6. Pressure valve

  7. Thermometer

jkacd-34-356f1.jpg
Table 1.
The averages and standard deviations of optical density of MTT.
Groups MTT1 Eosin2 MTT/Eosin3
Immediate control 3.01±0.25 0.49±0.11 6.45±1.46a
Cold preservation 0.81±0.26 0.42±0.11 2.08±0.94b
Slow freezing 1.62±0.24 0.47±0.13 3.62±0.88c
Slow freezing under pressure 1.94±0.44 0.44±0.14 4.71±1.24d

There were significant differences between groups with different subscripts(p<0.05).

The values of MTT1 are proportional to the vitality of the cells.

The values of Eosin2 are proportional to the area of the periodontal ligament cells.

So, the values of MTT/Eosin3 mean the vitality of the periodontal ligament cells per unit area.

Table 2.
The averages and standard deviations of optical density of WST-1.
Groups WST-11 Eosin2 WST-1/Eosin3
Immediate control 2.31±0.33 0.49±0.13 4.96±1.23a
Cold preservation 0.58±0.19 0.39±0.12 1.50±0.32b
Slow freezing 1.11±0.32 0.5±0.20 2.46±1.00c
Slow freezing under pressure 1.34±0.21 0.41±0.10 3.41±0.81d

There were significant differences between groups with different subscripts(p<0.05).

The values of WST-11 are proportional to the vitality of the cells.

The values of Eosin2 are proportional to the area of the periodontal ligament cells.

So the values of WST-1/Eosin3 mean the vitality of the periodontal ligament cells per unit area.

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    • Effects of Slow Programmable Cryopreservation on Preserving Viability of the Cultured Periodontal Ligament Cells from Human Impacted Third Molar
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      Journal of Korean Dental Science.2015; 8(2): 57.     CrossRef
    • The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure
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      THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS
      J Korean Acad Conserv Dent. 2009;34(4):356-363.   Published online January 14, 2009
      DOI: https://doi.org/10.5395/JKACD.2009.34.4.356
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    THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS
    Image
    Figure 1. Schematic diagram of program freezer with pressure vessel. Oxygen container : 3MPa of pressureProgram freezerPressure bottle2ml Cryotube: 1ml F medium + 10% DMSOTooth treated 10% DMSOPressure valveThermometer

    Figure 1.

    THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS
    Groups MTT1 Eosin2 MTT/Eosin3
    Immediate control 3.01±0.25 0.49±0.11 6.45±1.46a
    Cold preservation 0.81±0.26 0.42±0.11 2.08±0.94b
    Slow freezing 1.62±0.24 0.47±0.13 3.62±0.88c
    Slow freezing under pressure 1.94±0.44 0.44±0.14 4.71±1.24d
    Groups WST-11 Eosin2 WST-1/Eosin3
    Immediate control 2.31±0.33 0.49±0.13 4.96±1.23a
    Cold preservation 0.58±0.19 0.39±0.12 1.50±0.32b
    Slow freezing 1.11±0.32 0.5±0.20 2.46±1.00c
    Slow freezing under pressure 1.34±0.21 0.41±0.10 3.41±0.81d
    Table 1. The averages and standard deviations of optical density of MTT.

    There were significant differences between groups with different subscripts(p<0.05).

    The values of MTT1 are proportional to the vitality of the cells.

    The values of Eosin2 are proportional to the area of the periodontal ligament cells.

    So, the values of MTT/Eosin3 mean the vitality of the periodontal ligament cells per unit area.

    Table 2. The averages and standard deviations of optical density of WST-1.

    There were significant differences between groups with different subscripts(p<0.05).

    The values of WST-11 are proportional to the vitality of the cells.

    The values of Eosin2 are proportional to the area of the periodontal ligament cells.

    So the values of WST-1/Eosin3 mean the vitality of the periodontal ligament cells per unit area.

    Table 1.

    Table 2.

    Restor Dent Endod : Restorative Dentistry & Endodontics
    © The Korean Academy of Conservative Dentistry.
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