Computerized Reconstruction of Pulpal Blood Vessels Examined under Confocal Microscope

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1 / Kleoniki Lyroudia2 / Lucie Kubinova3 / Georgia Karayannopoulou4 / Ioannis Marras5 / Ioannis Pitas5

1Private Dentist, Thessaloniki, Greece
2Aristotle University of Thessaloniki, Department of Endodontology, Dental School, Thessaloniki, Greece
3Academy of Sciences of the Czech Republic, Institute of Physiology, Department of Biomathematics, Prague, Czech Republic
4Aristotle University of Thessaloniki, Medical School, Thessaloniki, Greece
5Aristotle University of Thessaloniki, Department of Informatics, Thessaloniki, Greece

Summary

The purpose of this study was the evaluation of 3 different histological methods for studying pulpal blood vessels in combination with 2 types of confocal microscope and computer assisted 3-dimensional reconstruction. 10 human, healthy, free of restorations or caries teeth that were extracted for orthodontic reasons were used. From these teeth, the pulp tissues of 5 were removed, fixed in formalin solution, dehydrated and embedded in paraffin. Serial cross sections 5μm thick were taken from 3 of the above mentioned pulpal tissues and stained with CD34 according to the immunohistochemical ABC technique, while the rest 2 were stained with CD34 and Cy5 by means of immunofluorescence after serial cross sectioning of 10μm. 5 of the 10 teeth were fixed, decalcified, serial cross sectioned (30μm thickness) and stained with eosin. The physical sections were examined under 2 types of confocal laser microscope. Serial images were taken for each section, alignment of the images was followed and finally 3-dimensional reconstructions of the pulpal vessels were achieved.

The combined use of immunofluorescence, confocal microscope and automatic segmentation proved to be a useful method for the detailed study of pulpal vasculature. The above method provides deep knowledge of the form and spatial relationship even of the smallest pulpal blood vessels with neighbouring structures like odontoblasts, which are essential for the fully understanding of their role and function within the dental pulp.

Keywords: CD34; Cy5; Immunofluorescence; 3D reconstruction

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Citation Information: Balkan Journal of Dental Medicine. Volume 19, Issue 1, Pages 43–49, ISSN (Online) 2335-0245, DOI: https://doi.org/10.1515/bjdm-2015-0033, July 2015

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