Abstract
ObjectivesThe aim of this investigation was the detailed analysis of the human pulp proteome using the new picosecond infrared laser (PIRL)-based sampling technique, which is based on a completely different mechanism compared to mechanical sampling. Proteome analysis of healthy pulp can provide data to define changes in the proteome associated with dental disease.Material and methodsImmediately after extraction of the entire, undamaged tooth, 15 wisdom teeth were deep frozen in liquid nitrogen and preserved at −80°C. Teeth were crushed, and the excised frozen pulps were conditioned for further analysis. The pulps were sampled using PIRL, and the aspirates digested with trypsin and analyzed with mass spectrometry. Pulp proteins were categorized according to their gene ontology terminus. Proteins identified exclusively in this study were searched in the Human Protein Atlas (HPA) for gaining information about the main known localization and function.ResultsA total of 1348 proteins were identified in this study. The comparison with prior studies showed a match of 72%. Twenty-eight percent of the proteins were identified exclusively in this study. Considering HPA, almost half of these proteins were assigned to tissues that could be pulp specific.ConclusionPIRL is releasing proteins from the dental pulp which are not dissolved by conventional sampling techniques.Clinical RelevanceThe presented data extend current knowledge on dental pulp proteomics in healthy teeth and can serve as a reference for studies on pulp proteomics in dental disease.
Highlights
Many cellular functions of the tooth are concentrated in the dental pulp [1, 2]
The differences in the number and composition of the identified dental pulp proteins from previous studies are in part due to different methods of determination as well as the presence of blood proteins, which can reduce the number of identifiable proteins because of the suppression effect in mass spectrometry (MS), which is in general a successful technique for protein analysis [8,9,10,11,12]
There is an overlap of proteins, which have been identified in all studies, which is similar in relationship to the number of proteins, identified in the individual studies
Summary
Many cellular functions of the tooth are concentrated in the dental pulp [1, 2]. The proteome represents the current functional status of a tissue. There are only a few studies dealing with the proteome analysis of dental pulp. The differences in the number and composition of the identified dental pulp proteins from previous studies are in part due to different methods of determination as well as the presence of blood proteins, which can reduce the number of identifiable proteins because of the suppression effect in mass spectrometry (MS), which is in general a successful technique for protein analysis [8,9,10,11,12].
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