Extraction and radioimmunoassay quantitation of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) from human dental pulp tissue

• Published in: Archives of oral biology Vol. 48; no. 3; pp. 249 - 254
• Main Authors: El-Karim, I, Lundy, F.T, Linden, G.J, Lamey, P.-J
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.03.2003
• Subjects: Acetic Acid; Adult; Chromatography, High Pressure Liquid - methods; Dental pulp; Dental Pulp - chemistry; Dentistry; Human; Humans; Neuropeptide Y; Neuropeptide Y - analysis; Neuropeptide Y - isolation & purification; Radioimmunoassay; Radioimmunoassay - methods; Reproducibility of Results; Specimen Handling - methods; Statistics, Nonparametric; Vasoactive Intestinal Peptide - analysis; Vasoactive Intestinal Peptide - isolation & purification; Vasoactive intestinal polypeptide; Human; Neuropeptide Y; Radioimmunoassay; Vasoactive intestinal polypeptide; Dental pulp
• ISSN: 0003-9969; 1879-1506
• DOI: 10.1016/S0003-9969(02)00213-3

The measurement of neuropeptides in complex biological tissue samples requires efficient and appropriate extraction methods so that immunoreactivity is retained for subsequent radioimmunoassay detection. Since neuropeptides differ in their molecular mass, charge and hydrophobicity, no single method will suffice for the optimal extraction of various neuropeptides. In this study, dental pulp tissue was obtained from 30 human non-carious teeth. Of the three different neuropeptide extraction methods employed, boiling in acetic acid in the presence of protease inhibitors yielded the highest levels of neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP). High pressure liquid chromatography (HPLC) analysis of dental pulp tissue verified the authenticity of the neuropeptides extracted.