Comparative immunohistochemical analysis of the peptidergic innervation of human primary and permanent tooth pulp

This immunohistochemical study sought to determine whether there are any differences in the peptidergic innervation of these pulps and whether dental caries is associated with changes in neuropeptide expression. Mandibular first permanent molars and second primary molars (n=120) were obtained from children requiring dental extractions under general anaesthesia. Extracted teeth were split longitudinally, placed in fixative, and categorized as intact, moderately carious or grossly carious. The coronal pulps were removed and 10-μm frozen sections were processed for indirect immunofluorescence. Double labelling employed combinations of the following antisera: (1) protein gene product 9.5, a general neuronal marker; (2) one of the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), galanin (GAL), enkephalin (ENK) and somatostatin (SOM). Image analysis was then used to determine the percentage area of immunostaining for each label within different anatomical regions of the coronal pulp. Sparse or absent immunoreactivity for GAL, ENK and SOM made analysis impossible. Analysis of CGRP, SP and VIP revealed significant interdentition differences, with their expression being significantly greater in permanent teeth, but this was not the case for NPY, with primary and permanent teeth demonstrating a similar amount of label for this peptide. Both dentitions showed significant increases in CGRP, SP, VIP and NPY expression with caries progression. These findings could have biological and clinical importance in connection with nociception, inflammation and healing.