J Korean Acad Conserv Dent.  2010 Jul;35(4):285-294. 10.5395/JKACD.2010.35.4.285.

The evaluation of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure

Affiliations
  • 1Department of Conservative Dentistry, College of Dentistry, Yonsei University, Seoul, Korea. sjlee@yuhs.ac
  • 2Department of Oral Pathology, College of Dentistry, Yonsei University, Seoul, Korea.
  • 3Department of Periodontology, College of Dentistry, Yonsei University, Seoul, Korea.
  • 4Department of Biotechnology, College of Life Science and Biothchnology, Yonsei University, Seoul, Korea.

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 0degrees C under pressure of 2 MPa), group 8 (low-temperature preservation at 0degrees C under no additional pressure), group 9 (low-temperature preservation at -5degrees C 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 37degrees C 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 (0degrees C/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 0degrees C under pressure of 2 MPa suggest the possibility for long term preservation of teeth.

Keyword

Periodontal ligament cell; MTT; WST-1; Viability; Low temperature preservation under pressure; Rapid freezing; Cryopreservation

MeSH Terms

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

Figure

  • 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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