J Korean Acad Conserv Dent.  2008 Jul;33(4):332-340. 10.5395/JKACD.2008.33.4.332.

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

• Ahn HJ
• Kim ES
• Kim J
• Kim DW
• Kim KY
• Lee CY
• Lee SJ

• KMID: 2176001
• DOI: http://doi.org/10.5395/JKACD.2008.33.4.332

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 4degrees C 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.

Keyword

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

MeSH Terms

Anesthesia
Animals
Cryopreservation
Dimethyl Sulfoxide
Eosine Yellowish-(YS)
Female
Humans
In Situ Nick-End Labeling
Magnetic Fields
Magnetics
Magnets
Molar
Periodontal Ligament
Rats
Tiletamine
Tooth
Dimethyl Sulfoxide
Eosine Yellowish-(YS)
Tiletamine

Figure

• 

  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).

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