Proteome Expression in Human Periodontal Ligament after Delayed Hypothermic Preservation

doi:10.1016/j.joen.2017.02.019

Journal of Endodontics

Volume 43, Issue 8, August 2017, Pages 1317-1322

https://doi.org/10.1016/j.joen.2017.02.019 Get rights and content

Highlights

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Vimentin and type VI collagen were the most prominent proteins in the periodontal ligament after delayed hypothermic preservation of teeth.
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The total size of the vimentin spots decreased gradually with increasing storage time, from 0 to 2 weeks, and decreased rapidly after dry storage.
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Only the dry storage group differed significantly from the immediately sampled group.

Abstract

Introduction

Previous occasional successes after delayed replantation suggest that the presence of viable cells may not be the only factor for successful periodontal regeneration in delayed replantation. Various other factors such as proteins or the extracellular matrix (ECM) may play a role in this process. The purpose of this study was to characterize changes in the proteome components of periodontal ligament (PDL) tissue after hypothermic preservation of the tooth.

Methods

Extracted teeth were divided into 4 groups: immediate sampling, sampling after 1 week of preservation at 4°C, sampling after 2 weeks of preservation at 4°C, and sampling after 1 week of dry storage at room temperature as a negative control. PDL tissues were collected from the root and stored immediately in liquid nitrogen. The tissues were subjected to 2-dimensional gel electrophoresis, and spot selection was executed. Selected spots that maintained the protein volume were then processed with matrix-assisted laser desorption and ionization time-of-flight mass spectrometry to identify the nature of the proteins.

Results

Thirty-five selected spots were analyzed. Sixteen spots were identified as vimentin, and 3 spots were type VI collagen. The size of the 16 vimentin spots decreased gradually with increasing storage time, from 0 to 2 weeks, and decreased rapidly after dry storage. However, only the dry storage group differed significantly from the immediately sampled group.

Conclusions

Vimentin was the most prominent protein followed by type VI collagen in volumetrically maintained protein spots. Although these proteins are known to be closely related with ECM integrity, the role of these proteins in delayed replantation is beyond the scope of this study. Further studies are needed to elucidate the possible role of these proteins for periodontal healing of delayed replantation.

Section snippets

Experimental Group Design and Tissue Sampling

Four volunteers were recruited from patients who were scheduled to have 4 premolars extracted in the Department of Orthodontics, Yonsei University Dental Hospital, Seoul, Korea. Informed consent to participate was obtained from each patient, and the research protocol was approved by the Institutional Review Board of Yonsei University Dental Hospital (number 2-2013-0033).

All extraction and sampling procedures were performed by 1 operator at the Department of Conservative Dentistry, Yonsei

2-DE and Image Analysis

2-DE images were obtained for each of the 4 experimental groups (Fig. 1A). The overall spot patterns were similar, implying that the sampling and analysis procedures were reliable. About 1300 spots were detected on each gel, and 120 spots changed by more than 2-fold in the experimental groups compared with the immediate group.

Initial Spot Selection for Protein Identification

Twenty-seven spots were selected according to the conditions outlined in the Materials and Methods section.

MALDI-TOF Protein Identification

Eleven of the 27 initially selected spots were identified as

Discussion

Vimentin was the most notable protein expressed in this study. This protein appeared consistently under hypothermic storage conditions, but its levels decreased rapidly in dry storage conditions. Vimentin is a type III intermediate filament protein that constructs a cytoskeletal system. Vimentin filaments, which are associated with collagen messenger RNAs (mRNAs) in a 5′SL- and La-related protein (LARP6)-dependent manner, act to stabilize collagen mRNAs (15). The increased stability of collagen

Acknowledgments

The authors thank Yonsei Proteome Research Center (www.proteomix.org) for technical support for proteomic analysis.

Supported by a faculty research grant of Yonsei University College of Dentistry for 2013 (grant no. 6-2013-0131).

The authors deny any conflicts of interest related to this study.

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View full text

Basic ResearchProteome Expression in Human Periodontal Ligament after Delayed Hypothermic Preservation

Highlights

Abstract

Introduction

Methods

Results

Conclusions

Section snippets

Experimental Group Design and Tissue Sampling

2-DE and Image Analysis

Initial Spot Selection for Protein Identification

MALDI-TOF Protein Identification

Discussion

Acknowledgments

Arch Oral Biol

Biomaterials

Biomaterials

Biotechnol Adv

J Mol Biol

Biomaterials

Biomaterials

Replantation of 400 avulsed permanent incisors. 4. Factors related to periodontal ligament healing

Endod Dent Traumatol

The effect of limited drying or removal of the periodontal ligament. Periodontal healing after replantation of mature permanent incisors in monkeys

Acta Odontol Scand

International Association of Dental Traumatology guidelines for the management of traumatic dental injuries: 2. Avulsion of permanent teeth

Dent Traumatol

Delayed replantation of an avulsed maxillary premolar with open apex: a 24 months follow-up case report

J Indian Soc Pedod Prev Dent

Basic Research
Proteome Expression in Human Periodontal Ligament after Delayed Hypothermic Preservation