Quantification of neural protein in extirpated tooth pulp

Because the pulp tissue extirpated during root canal procedures might serve as a valuable resource with which to assess underlying mechanisms of persistent pain, we sought to determine whether standard Western blotting techniques could be used to quantify neural proteins in pulp extirpated from teeth with irreversible pulpitis. Pulp harvested from healthy intact teeth extracted for orthodontic reasons was used for comparison. The neural marker PGP9.5 was detectable in all samples tested. A membrane enrichment protocol enabled detection of even low abundance, high molecular weight proteins such as the sodium channel alpha-subunit NaV1.8. Importantly, it was possible to quantify a approximately 6-fold increase in the relative density of NaV1.8 in inflamed pulp compared with control pulp. Our results suggest that it should be possible to use extirpated tooth pulp to validate mechanisms of persistent pain implicated in preclinical studies as well as evaluate the therapeutic efficacy of novel antinociceptive interventions.