The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure
Article information
Restor Dent Endod. 2010;35(4):285-294
Publication date (electronic) : 2010 July 31
doi :
https://doi.org/10.5395/JKACD.2010.35.4.285
Received 2010 May 29; Revised 2010 June 19; Accepted 2010 July 03.
Abstract
The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group.
Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at 0℃ under pressure of 2 MPa), group 8 (low-temperature preservation at 0℃ under no additional pressure), group 9 (low-temperature preservation at -5℃ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in 37℃ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD 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 7 (0℃/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance).
By the results of this study, low-temperature preservation at 0℃ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.
Keywords: Periodontal ligament cell; MTT; WST-1; Viability; Low temperature preservation under pressure; Rapid freezing; Cryopreservation
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