Journal List > J Korean Acad Conserv Dent > v.35(4) > 1056421

Chung, Kim, Choi, Kim, Park, and Lee: The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure

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.

Figures and Tables

Figure 1
Schematic diagram of program freezer with pressure vessel.
a. Oxygen container: 2 or 3 MPa of pressure, b. Program freezer, c. Pressure bottle, d. 2 ml Cryotube: 1 ml F medium + 10% DMSO, e. Tooth treated 10% DMSO, f. Pressure valve, g. Thermometer.
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Table 1
Experimental Groups
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Table 2
The averages and standard deviations of optical density of MTT
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a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

Table 3
The averages and standard deviations of optical density of WST-1
jkacd-35-285-i003

a,b,c,d,e,f: Different superscript letters means statistical difference (p < 0.05).

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