THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS

Young-Eun Lee; Eui-Seong Kim; Jin Kim; Seung-Hoon Han; Seung-Jong Lee

doi:10.5395/JKACD.2009.34.4.356

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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 Lee^1,*; Journal of Korean Academy of Conservative Dentistry 2009;34(4):356-363.
DOI: https://doi.org/10.5395/JKACD.2009.34.4.356
Published online: January 14, 2009

¹Department of Conservative Dentistry, College of Dentistry, Yonsei University

²Department of Oral Pathology, Oral Cancer Research Center, Yonsei University

³Department of Conservative Dentistry, Kangbuk Samsung Hospital

*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

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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Abstract
REFERENCES
참고문헌

Abstract

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.

Oxygen container : 3MPa of pressure
Program freezer
Pressure bottle
2ml Cryotube: 1ml F medium + 10% DMSO
Tooth treated 10% DMSO
Pressure valve
Thermometer

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±1.46^a
Cold preservation	0.81±0.26	0.42±0.11	2.08±0.94^b
Slow freezing	1.62±0.24	0.47±0.13	3.62±0.88^c
Slow freezing under pressure	1.94±0.44	0.44±0.14	4.71±1.24^d

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±1.23^a
Cold preservation	0.58±0.19	0.39±0.12	1.50±0.32^b
Slow freezing	1.11±0.32	0.5±0.20	2.46±1.00^c
Slow freezing under pressure	1.34±0.21	0.41±0.10	3.41±0.81^d

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.

REFERENCES

참고문헌

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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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Figure

THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS

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±1.46^a
Cold preservation	0.81±0.26	0.42±0.11	2.08±0.94^b
Slow freezing	1.62±0.24	0.47±0.13	3.62±0.88^c
Slow freezing under pressure	1.94±0.44	0.44±0.14	4.71±1.24^d

Groups	WST-1¹	Eosin²	WST-1/Eosin³
Immediate control	2.31±0.33	0.49±0.13	4.96±1.23^a
Cold preservation	0.58±0.19	0.39±0.12	1.50±0.32^b
Slow freezing	1.11±0.32	0.5±0.20	2.46±1.00^c
Slow freezing under pressure	1.34±0.21	0.41±0.10	3.41±0.81^d

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.