Tissue-engineered oral epithelial barrier for dental material testing: towards establishing in vitro biomimetic models - Underlying data

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.

本数据集为论文《用于牙科材料测试的组织工程口腔上皮屏障：构建体外仿生模型》（https://doi.org/10.1089/ten.tec.2024.0154）的配套CT数据。

作者：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 Fatouros b, c, Athina Bakopoulou a

a 希腊塞萨洛尼基亚里士多德大学健康科学学院牙科学院修复牙科系组织工程核心实验室，塞萨洛尼基 54124

b 希腊塞萨洛尼基亚里士多德大学健康科学学院药学院制药技术系，塞萨洛尼基 54124

c 希腊塞萨洛尼基亚里士多德大学跨学科研究与创新中心（CIRI-AUTH），塞萨洛尼基 57001

d 希腊塞萨洛尼基亚里士多德大学兽医学院解剖与组织学实验室，塞萨洛尼基 54124

e 希腊塞萨洛尼基亚里士多德大学健康科学学院牙科学院口腔医学/病理学系，塞萨洛尼基 54124

f 希腊塞萨洛尼基亚里士多德大学理学院生物学院植物学系，塞萨洛尼基 54124

g 英国南安普顿大学工程与环境学院μ-VIS X射线成像中心，南安普顿 SO17 1BJ

h 英国南安普顿大学生命科学研究所，南安普顿 SO17 1BJ

i 法国艾克斯-马赛大学运动科学研究所法国国家科学研究中心，马赛 13385

### 组织工程口腔上皮（TEOE）厚度测量
X射线显微计算机断层扫描（μCT）被用于检测石蜡包埋的组织工程口腔上皮（TEOE）的微观结构，可通过体积分析实现全面的三维厚度评估（成像参数详见补充材料）(11)。扫描采用各向同性体素边长为6.0 μm的设置。使用Dragonfly软件中的“体积厚度图”工具完成三维局部厚度分析（详见补充材料），实现组织厚度空间分布与变异的可视化及定量分析。

成像实验在南安普顿大学μ-VIS X射线成像中心（https://muvis.org）/3D X射线组织学平台开展，使用基于尼康XTH225ST系统（英国尼康计量公司）定制的μCT扫描仪，该设备针对精细组织学分析进行了优化(1)。扫描参数设置为80 kVp / 86 μA（6.88 W），源物距37.5 mm，源探测器距937.4 mm，由此获得25倍放大倍率与6.0 μm的各向同性体素边长。图像采集过程中，使用2850×2850像素的探测器收集4001幅投影图像，每幅投影平均4帧，单幅投影曝光时间为500 ms。

重建数据集的可视化与分析使用Dragonfly软件（加拿大彗星科技有限公司；软件获取地址：http://www.theobjects.com/dragonfly）完成。在对组织层进行分割后，通过该软件的“体积厚度图”工具开展三维局部厚度评估。局部厚度分析可实现组织工程口腔上皮（TEOE）内部厚度空间分布与变异的可视化及定量表征。

研究采用局部厚度直方图可视化手段，阐明TEOE整体的厚度分布特征。此类直方图可直观展示特定截面厚度对应的体素数量，既以图形化方式呈现组织样本的厚度变异情况，又可定量给出样本的平均厚度。

值得注意的是，该技术的体积成像与非破坏性特性支持整块组织成像，这对解决组织样本收缩带来的挑战至关重要。样本收缩是固定过程中脱水导致的后果，部分样本会出现卷曲现象。尽管卷曲并非常见情况，但本分析方法可用于评估形状复杂的样本，例如图4中展示的卷曲样本。与传统二维方法（如经典组织学）依赖切片角度、且在非完美垂直切片时存在局限不同，基于μCT的X射线组织学技术可分析所有样本，包括形状复杂的标本。