DOI: 10.5281/zenodo.11181663
title: Tissue-engineered oral epithelial barrier for dental material testing: towards establishing in vitro biomimetic models - Underlying data

description: Underlying CT data of "Tissue-engineered oral epithelial barrier for dental material testing: towards establishing in vitro biomimetic models"https://doi.org/10.1089/ten.tec.2024.0154

Foteini Machla a, Paraskevi Kyriaki Monou b, c, Chrysanthi Bekiari d, Dimitrios Andreadis e Evangelia Kofidou d, , Emmanouel Panteris f, Orestis L. Katsamenis g, h, Maria Kokoti a, Petros Koidis a, Imad About i, Dimitrios Fatourosb, c, Athina Bakopoulou a

a Department of Prosthodontics, Tissue Engineering Core Unit, School of Dentistry, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece

b Department of Pharmaceutical Technology, School of Pharmacy, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece

c Center for Interdisciplinary Research and Innovation (CIRI-AUTH), Thessaloniki 57001, Greece

d Laboratory of Anatomy and Histology, Veterinary School, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece

e Department of Oral Medicine/Pathology, School of Dentistry, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece

f Department of Botany, School of Biology, Faculty of Sciences, Aristotle University of Thessaloniki, Thessaloniki 54124, Greece

g μ-VIS X-ray Imaging Centre, Faculty of Engineering and the Environment, University of Southampton, Southampton SO17 1BJ, United Kingdom

h Institute for Life Sciences, University of Southampton, Southampton SO17 1BJ, United Kingdom UK

i Centre National de la Recherche Scientifique, Institute of Movement Sciences, Aix Marseille University, Marseille 13385, France

 

Measurement of ΤΕΟΕ thickness

X-ray computed micro-tomography (μCT) as employed to examine the microstructure of the paraffin-embedded tissue engineered oral epithelium (TEOE), enabling comprehensive 3D assessment of thickness using volumetric analysis (cf. supplementary for imaging parameters) (11). Imaging was conducted using an isotropic voxel-edge of 6.0 μm. Local Thickness was carried out in 3D using the “Volume Thickness Map” tool within Dragonfly software (cf. supplementary), allowing for the visualization and quantification of the spatial distribution and variability of tissue thickness.

Imaging was conducted at the University of Southampton’s μ-VIS X-ray Imaging Centre ( https://muvis.org ) / 3D X-ray Histology facility, utilizing a customised μCT scanner optimisedfor intricate histological analyses(1), based on Nikon’s XTH225ST system (Nikon Metrology, UK). Operating parameters were set at 80 kVp / 86 μA (6.88 W), with a source-to-object distance of 37.5 mm and a source-to-detector distance of 937.4 mm, resulting in a magnification factor of 25x and an isotropic voxel-edge of 6.0 μm. Imaging acquisition involved the collection of 4001 projections using a 2850 x 2850 dexels detector, by averaging 4 frames per projection, with an exposure time of 500 ms per projection.

Visualisation and analysis of the reconstructed dataset was done using Dragonfly software (Comet Technologies Canada Inc.; software accessible at http://www.theobjects.com/dragonfly). Assessment of Local Thickness was carried out in 3D using the “Volume Thickness Map” tool within Dragonfly software, following segmentation of the tissue layer. Local thickness analysis allowed for the visualization and quantification of the spatial distribution and variability of thickness within the tissue engineered oral epithelium (TEOE).

Visual representation of local thickness histograms was employed to elucidate the distribution of thickness throughout the TEOE. These histograms effectively illustrate the number of voxels associated with specific cross-sectional thickness, offering both a graphical depiction of the variation in thickness across the tissue sample, and a quantitative measure of the average thickness of the specimen.

It's worth noting, the volumetric and non-destructive nature of the technique enabled whole-block imaging, which proved crucial in addressing challenges arising from tissue sample shrinkage. This shrinkage, a consequence of dehydration during the fixation process, can occur in some cases and lead led to the specimen wrapping. While wrapping is not a common occurrence, this analysis method allowed for the evaluation of challenging-shaped specimens, such as the wrapped one presented in Figure 4. Unlike conventional 2D methods such as classical histology, which rely on the angle of slicing and encounter limitations when dealing with non-perfectly perpendicular slicing, μCT-based XRH enables analysis of all specimens, including those with complex shapes.

 
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Figure-XCTv2_mucosa.pngFigure shown in the paper

Figure-Sxv2_mucosa.pngSupplementary image providing additional views of the mucosa culture.

sc_3DProd-of-7xMed_22220_20240313_XRH_3922_OLK_XTAC12147-FFPE_974x670x1347x16bit.rawEnhanced microCT volume data, generated by applying a 7-slice moving average to enhance SNR; size metadata needed for importing into visualization software shown at the end of the file (XxYxZxBitDepth)

LocakThickness.opjuOrigin project file analysing porosity in the dataset.

22220_20240313_XRH_3922_OLK_XTAC12147-FFPE_2487x1416x2850x16bit.rawOriginal microCT volume data with size metadata needed for importing into visualization software shown at the end of the file