THE EFFICACY OF PROGRAMMED CRYO-PRESERVATION UNDER PRESSURE IN RAT PERIODONTAL LIGAMENT CELLS
Article information
Restor Dent Endod. 2009;34(4):356-363
Publication date (electronic) : 2009 January 14
doi :
https://doi.org/10.5395/JKACD.2009.34.4.356
Received 2009 April 23; Revised 2009 June 16; Accepted 2009 June 22.
Abstract
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
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
The averages and standard deviations of optical density of MTT.
The averages and standard deviations of optical density of WST-1.
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