Evaluation of the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field

Article information

Restor Dent Endod. 2008;33(4):332-340

Publication date (electronic) : 2008 July 31

doi : https://doi.org/10.5395/JKACD.2008.33.4.332

Hyun-Jung Ahn , Eui-Seong Kim , Jin Kim , Duck-Won Kim , Ki-Yeol Kim , Chan-Young Lee , Seung-Jong Lee

Department of Conservative Dentistry, Yonsei University, Seoul, Korea.

Corresponding Author: Eui-Seong Kim. Department of Conservative Dentistry, College of Dentistry, Yonsei University, 134 Shinchon-Dong, Seodaemun-Ku, Seoul, 120-752, Korea. Tel: 82-2-2228-8700, Fax: 82-2-313-7575, andyendo@yuhs.ac

Received 2008 April 14; Revised 2008 May 20; Accepted 2008 May 21.

Abstract

The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4℃ for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2.

From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.

Keywords: TUNEL; Periodontal ligament cell; MTT; Magnetic field; Cryopreservation

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Figure 1

MTT/Eosin ratio.

1: immediately extraction

2: cold preservation at 4℃

3: rapid cryopresevation in liquid nitrogen

4: slow cryopreservation with magnetic field of 1G

5: slow cryopreservation

★: p < 0.05.

Figure 2

Average and standard deviation of percentage of positive cells by TUNEL test.

1: immediately extraction

2: cold preservation at 4℃

3: rapid cryopresevation in liquid nitrogen

4: slow cryopreservation with magnetic field of 1G

5: slow cryopreservation

★: p < 0.05.

Figure 3

Micrograph of experimental groups by TUNEL test. Arrow shows positive cell.

A. immediately extraction (× 400).

B. cold preservation (× 400).

C. rapid cryopreservation (× 400).

D. slow cryopreservation with magnetic field (× 400).

E. slow cryopreservation (× 400).