title: Immunocytochemical investigation of neurovascular relationships in human tooth pulp.
abstract: This study sought to explore the anatomical relationships between peptidergic nerves and blood vessels within human primary and permanent teeth. Extracted primary and permanent molars (n = 120) were split longitudinally, placed in Zamboni's fixative and the coronal pulps were processed for indirect immunofluorescence. Ten-micrometre-thick serial frozen pulp sections were triple-labelled using combinations of the following antisera: (1) protein gene-product 9.5 (PGP 9.5), a general neuronal marker; (2) one of the neuropeptides, calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP) or neuropeptide Y (NPY); and (iii) the lectin Ulex europeus, a label for vascular endothelium. The mid-coronal pulp region was examined, using fluorescence microscopy, to determine the proportion of blood vessels showing a positive innervation (recorded when PGP 9.5-labelled nerves appeared to intersect the vessel wall). In addition, the percentage of these vascular-related nerves expressing each of the above neuropeptides was recorded. Overall, 20% of pulpal blood vessels appeared to have a positive innervation. In the main these were thick-walled arterioles. Capillaries, venules and lymphatics were mostly devoid of an associated innervation. Ninety-two per cent of vascular-related nerves expressed CGRP, 87% expressed SP, 15% expressed VIP and 80% expressed NPY. There were no significant differences in overall innervation or peptide-related innervation between primary and permanent teeth (P < 0.05, ANOVA), indicating that pulpal blood flow is likely to be subject to similar neurological control mechanisms in both dentitions.
authors.affiliation: School of Clinical Dentistry, University of Sheffield, UK. h.d.rodd@sheffield.ac.uk
authors.forename: Helen D
authors.initials: HD
authors.lastname: Rodd
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Boissonade
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis; D003782:Dental Pulp / Q000033:anatomy & histology* / Q000098:blood supply / Q000294:innervation; D006651:Histocytochemistry / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008856:Microscopy, Fluorescence; D008963:Molar; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009478:Neuropeptide Y / Q000032:analysis; D013373:Substance P / Q000032:analysis; D013869:Thiolester Hydrolases / Q000032:analysis; D014094:Tooth, Deciduous; D043222:Ubiquitin Thiolesterase; D014660:Vasoactive Intestinal Peptide / Q000032:analysis
chemical_list: D015415:Biomarkers; D009419:Nerve Tissue Proteins; D009478:Neuropeptide Y; D013373:Substance P; D014660:Vasoactive Intestinal Peptide; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase; D015740:Calcitonin Gene-Related Peptide