J Korean Acad Conserv Dent.  2009 Jul;34(4):356-363. 10.5395/JKACD.2009.34.4.356.

The efficacy of programmed cryo-preservation under pressure in rat periodontal ligament cells

• Lee YE
• Kim ES
• Kim J
• Han SH
• Lee SJ

• KMID: 2176168
• DOI: http://doi.org/10.5395/JKACD.2009.34.4.356

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 4degrees 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 37degrees 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.

Keyword

periodontal ligament cell; MTT; WST-1; viability; pressure; cryo-preservation

MeSH Terms

Anesthesia
Animals
Baths
Dimethyl Sulfoxide
Eosine Yellowish-(YS)
Female
Freezing
Humans
Molar
Periodontal Ligament
Rats
Tiletamine
Tooth
Dimethyl Sulfoxide
Eosine Yellowish-(YS)
Tiletamine

Figure

• 

  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

Reference

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