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Odontoblast RNA stability in different temperature‐based protocols for tooth storage
Author(s) -
Conde M. C. M.,
Nedel F.,
Campos V. F.,
Smith A. J.,
Nör J. E.,
Demarco F. F.,
Tarquinio S. B. C.
Publication year - 2012
Publication title -
international endodontic journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.988
H-Index - 119
eISSN - 1365-2591
pISSN - 0143-2885
DOI - 10.1111/j.1365-2591.2011.01971.x
Subject(s) - trizol , odontoblast , rna extraction , human tooth , rna , pulp (tooth) , housekeeping gene , molar , ribosomal rna , dentistry , chemistry , gene expression , enamel paint , medicine , gene , biochemistry
Conde MCM, Nedel F, Campos VF, Smith AJ, Nör JE, Demarco FF, Tarquinio SB. Odontoblast RNA stability in different temperature‐based protocols for tooth storage. International Endodontic Journal , 45 , 266–272, 2012. Abstract Aim To evaluate the effect of four tooth storage temperature‐based methods on quality of RNA obtained from cells retrieved from human dental pulps and human pre‐dentine. Methodology RNA was isolated from dental pulp tissue and from cells retrieved by scraping the pre‐dentine of freshly extracted human third molars ( n = 15) using TRIzol ® reagent. Teeth were randomly assigned to the following temperature conditions: immediate RNA isolation after tooth extraction, liquid nitrogen (24 h), −80 °C (24 h), 20 °C (24 h) and 4 °C (6 h). RNA integrity was checked by the density of 28S and 18S ribosomal RNA. RT‐PCR was used to analyse the expression of odontoblast makers (DSPP, DMP1 and MEPE) and the housekeeping gene GAPDH. Results All experimental conditions evaluated preserved RNA integrity. The three odontoblastic markers were amplified from the pulp tissue and from the cells associated with pre‐dentine. Conclusion The four storage options allowed RNA isolation for RT‐PCR analysis. These findings may facilitate the use of clinically derived human dental pulp and odontoblasts for endodontic research.