Evaluation of periodontal ligament cell viability in rat teeth according to various extra-oral dry storage times using MTT assay

In-Soo Jeon; Eui-Seong Kim; Jin Kim; Seung-Jong Lee

doi:10.5395/JKACD.2006.31.5.398

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Original Article Evaluation of periodontal ligament cell viability in rat teeth according to various extra-oral dry storage times using MTT assay: In-Soo Jeon¹, Eui-Seong Kim¹, Jin Kim^1,2, Seung-Jong Lee^1,2; Journal of Korean Academy of Conservative Dentistry 2006;31(5):398-408.
DOI: https://doi.org/10.5395/JKACD.2006.31.5.398
Published online: September 30, 2006

¹Department of Conservative Dentistry, College of Dentistry, Yonsei University, Seoul, Korea.

²Oral Cancer Research Institute, College of Dentistry, Yonsei University, Seoul, Korea.

³Oral Science Research Center, College of Dentistry, Yonsei University, Seoul, Korea.

Corresponding Author: Seung-Jong Lee. Department of Conservative Dentistry, College of Dentistry, Yonsei University, 134, Shinchon-Dong, Seodaemun-Ku, Seoul, 120-752, Korea. Tel: 82-2-2228-8700, Fax: 82-2-313-7575, sjlee@yumc.yonsei.ac.kr

• Received: June 26, 2006 • Revised: August 9, 2006 • Accepted: September 11, 2006

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Abstract
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Abstract

The purpose of this study was to verify the usefulness of MTT analysis as a tool of measurement of the periodontal ligament cell viability from the extracted rat molar.

A total of 80 Sprague-Dawley white female rat of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted under Ketamine anesthesia. Twenty-four teeth of each group (divided as five groups depending upon the time-lapse after extraction such as immediate, 10, 20, 40 and 60 minutes) were immersed in 200 µl of MTT solution (0.5 mg/ml) and processed for optical density measurements . Another 10 teeth of each group were treated as same as above and sectioned at 10 µm for microscopic examination.

All measurements values were divided by the value of hematoxylin-eosin staining which represented the volume of each corresponding samples. Immediate and 10 minute groups showed highest MTT values followed by 20, 40, and 60 minutes consecutively. Statistical significance (p < 0.05) existed between all groups except in immediate versus 10 minute groups and 40 versus 60 minutes. Histological findings also showed similar findings with MTT results in crystal shape and crystal numbers between the experimental groups.

These data indicate that in vivo MTT analysis may be of value for evaluation of the periodontal ligament cell viability without time- consuming cell culturing processes.
Keywords: Periodontal ligament cell; Viability; MTT analysis; Preservation

REFERENCES

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Figure 1

Optical density of MTT and H-E staining following different extra-oral time (min) in Maxillary. 1st molar.

Figure 2

Optical density of MTT and H-E staining following different extra-oral time (min) in Maxillary. 2nd molar.

Figure 3

Statistical significance from Optical density of MTT and H-E staining following different extra-oral time (min) in Maxillary. 1st plus 2nd molar.

Figure 4

Serial section view from apex to cervicalportion in the immediate group (× 100).

Figure 5

Serial section view from apex to cervicalportion in the 60 minute time - lapse group (× 100).

Figure 6

Representative view after MTT staining in the immediate group.

Figure 7

Representative view after MTT staining in the 10 minute time - lapse group.

Figure 8

Representative view after MTT staining in the 20 minute time - lapse group.

Figure 9

Representative view after MTT staining in the 40 minute time - lapse group.

Figure 10

Representative view after MTT staining in the 60 minute time - lapse group.

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Evaluation of periodontal ligament cell viability in rat teeth according to various extra-oral dry storage times using MTT assay

J Korean Acad Conserv Dent. 2006;31(5):398-408. Published online September 30, 2006

DOI: https://doi.org/10.5395/JKACD.2006.31.5.398

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Figure

Evaluation of periodontal ligament cell viability in rat teeth according to various extra-oral dry storage times using MTT assay

Figure 1 Optical density of MTT and H-E staining following different extra-oral time (min) in Maxillary. 1st molar.

Figure 2 Optical density of MTT and H-E staining following different extra-oral time (min) in Maxillary. 2nd molar.

Figure 3 Statistical significance from Optical density of MTT and H-E staining following different extra-oral time (min) in Maxillary. 1st plus 2nd molar.

Figure 4 Serial section view from apex to cervicalportion in the immediate group (× 100).

Figure 5 Serial section view from apex to cervicalportion in the 60 minute time - lapse group (× 100).

Figure 6 Representative view after MTT staining in the immediate group.

Figure 7 Representative view after MTT staining in the 10 minute time - lapse group.

Figure 8 Representative view after MTT staining in the 20 minute time - lapse group.

Figure 9 Representative view after MTT staining in the 40 minute time - lapse group.

Figure 10 Representative view after MTT staining in the 60 minute time - lapse group.

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Evaluation of periodontal ligament cell viability in rat teeth according to various extra-oral dry storage times using MTT assay