Published January 15, 2021 | Version v1
Journal article Open

Using Histological Staining Techniques to Improve Visualization and Interpretability of Tooth Cementum Annulation Analysis

  • 1. University of Novi Sad, Faculty of Medicine, Novi Sad, Republic of Serbia
  • 2. University of Novi Sad, Biosense Institute, Novi Sad, Republic of Serbia
  • 3. University of Novi Sad, Biosense Institute, Novi Sad, Republic of Serbia. 3 University of Belgrade, Faculty of Philosophy, Belgrade, Republic of Serbia and University of Belgrade, Faculty of Philosophy, Belgrade, Republic of Serbia

Description

The aim of the present study was to evaluate the effect of different staining techniques on applicability and accuracy of tooth cementum annulation (TCA) method. Nine decalcination techniques, 8 dehydration protocols and 8 different techniques were applied in 3 teeth from the persons of a known age. Black and white, and color images of histological sections were captured. An xray was taken of each tooth and they were photographed. Researchers were asked to observe both black/white and color images of histological sections. Researchers were divided into two groups. The first group analyzed histological images only, and the second group had photos of teeth and X-rays. In the first group of observers (without X ray) the differences in age estimation between real and observed age were significant for 2 younger patients, but not for the oldest patient, where the observed and real values matched. Of the 6 raters, the assesments of the last 3 (that used x-ray images together with histological sections) did not differ significantly from the real values. Extensive analysis and multiple repetitions performed in the present investigation revealed that the most optimal method of decalcification for TCA method was EDTA II for a period longer than 14 days at a section thickness of 2-3mm, while the most optimal protocol for dehydration was number IV. When it comes to staining, the most optimal staining protocol used for the cemental lines visualization and counting was Crocein Scarlet/Acid Fuchsin staining and Toluidine blue staining used at semithin section. Additional use of preexperimental evaluation employing x-ray of analyzed teeth decreased the errors of age estimation.

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Using Histological Staining Techniques to Improve Visualization and Interpretability of Tooth Cementum Annulation Analysis.pdf

Additional details

Funding

Bioarchaeology of Ancient Europe: People, Animals and Plants in the Prehistory of Serbia 47001
Ministry of Education, Science and Technological Development
BIRTH – Births, mothers and babies: prehistoric fertility in the Balkans between 10000 – 5000 BC 640557
European Commission