The study of molecular composition in biomimetic interface of biocomposite/dentin

The aim of the study is the problem of formation of the biomimetic interface between the dental product and dentin of the human tooth as well as the investigations of molecular-chemical features in biointerface with the use of molecular multivariate IR-visualization. The data on synchrotron IR-mapping made enabled to differentiate the regions of sound dentin tissue and biomimetic transition layer and also to determine molecular groups responsible for the process of integration.

A bilayered tissue engineered in vitro model simulating the tooth periodontium

Due to the complexity of the structure of the tooth periodontium, regeneration of the full tooth attachment is not a trivial task. There is also a gap in models that can represent human tooth attachment in vitro and in vivo. The aim of this study was to develop a bilayered in vitro construct that simulated the tooth periodontal ligament and attached alveolar bone, for the purpose of tissue regeneration and investigation of physiological and orthodontic loading. Two types of materials were used to develop this construct: sol-gel 60S10Mg derived scaffold, representing the hard tissue component of the periodontium, and commercially available Geistlich Bio-Gide® collagen membrane, representing the soft tissue component of the tooth attachment. Each scaffold was dynamically seeded with human periodontal ligament cells (HPDLCs). Scaffolds were either cultured separately, or combined in a bilayered construct, for 2 weeks. Characterisation of the individual scaffolds and the bilayered constructs included biological characterisation (cell viability, scanning electron microscopy to confirm cell attachment, gene expression of periodontium regeneration markers), and mechanical characterisation of scaffolds and constructs. HPDLCs enjoyed a biocompatible 3-dimensional environment within the bilayered construct components. There was no drop in cellular gene expression in the bilayered construct, compared to the separate scaffolds.

A single-cell interactome of human tooth germ from growing third molar elucidates signaling networks regulating dental development

Background Development of dental tissue is regulated by extensive cell crosstalk based on various signaling molecules, such as bone morphogenetic protein (BMP) and fibroblast growth factor (FGF) pathways. However, an intact network of the intercellular regulation is still lacking. Result To gain an unbiased and comprehensive view of this dental cell interactome, we applied single-cell RNA-seq on immature human tooth germ of the growing third molar, discovered refined cell subtypes, and applied multiple network analysis to identify the central signaling pathways. We found that immune cells made up over 80% of all tooth germ cells, which exhibited profound regulation on dental cells via Transforming growth factor-β, Tumor necrosis factor (TNF) and Interleukin-1. During osteoblast differentiation, expression of genes related to extracellular matrix and mineralization was continuously elevated by signals from BMP and FGF family. As for the self-renewal of apical papilla stem cell, BMP-FGFR1-MSX1 pathway directly regulated the G0-to-S cell cycle transition. We also confirmed that Colony Stimulating Factor 1 secreted from pericyte and TNF Superfamily Member 11 secreted from osteoblast regulated a large proportion of genes related to osteoclast transformation from macrophage and monocyte. Conclusions We constructed the intercellular signaling networks that regulated the essential developmental process of human tooth, which served as a foundation for future dental regeneration engineering and the understanding of oral pathology.

In Vivo Evaluation of Decellularized Human Tooth Scaffold for Dental Tissue Regeneration

Conventional root canal treatment may result in loss of tooth vitality, which can lead to unfavorable treatment outcomes. Notably, a ceased tooth development of immature permanent teeth with open apices, regeneration of periodontal ligaments (PDL), and pulp is highly expected healing process. For regeneration, the scaffold is one of the critical components that carry biological benefits. Therefore, this study evaluated a decellularized human tooth as a scaffold for the PDL and pulp tissue regeneration. A tooth scaffold was fabricated using an effective decellularization method as reported in previous studies. PDL stem cells (PDLSCs) and dental pulp stem cells (DPSCs) obtained from human permanent teeth were inoculated onto decellularized scaffolds, then cultured to transplant into immunosuppressed mouse. After 9 weeks, PDLSCs and DPSCs that were inoculated onto decellularized tooth scaffolds and cultured in an in vivo demonstrated successful differentiation. In PDLSCs, a regeneration of the cementum/PDL complex could be expected. In DPSCs, the expression of genes related to revascularization and the hard tissue regeneration showed the possibility of pulp regeneration. This study suggested that the potential possible application of decellularized human tooth could be a scaffold in regeneration PDL and pulp tissue along with PDLSCs and DPSCs, respectively, as a novel treatment method.

Differences in the stemness characteristics and molecular markers of distinct human oral tissue neural crest-derived multilineage cells

Background Although multilineage cells derived from oral tissue, especially the dental pulp, apical papilla, periodontal ligament, and oral mucosa, have neural crest-derived stem cell (NCSC)-like properties, the differences in the characteristics of these progenitor cell compartments remain unknown. The primary aim of the current study was to elucidate these differences. Methods Multilineage sphere-forming apical papilla-derived cells (APDCs), periodontal ligament-derived cells (PDLDCs), and oral mucosa stroma-derived cells (OMSDCs) from the same individuals were isolated from impacted developing teeth. All sphere-forming cells were characterized by biological analyses of stem cells. Additionally, composites of these cells and multiporous hydroxyapatite scaffolds were transplanted into immunocompromised mice. Results All sphere-forming cells expressed neural crest-related markers. Although APDCs and PDLDCs showed greater mineralized-cell differentiation, they exhibited poorer differentiation into adipocytes in vitro than OMSDCs. In immunocompromised mice, APDCs were better able to form hard tissues than PDLDCs and OMSDCs. Moreover, the expression of certain tissue-specific markers, such as CD24 and CD56 (NCAM1), differed among the tissue-derived cells. Surprisingly, the expression of only CD24 and CD56 could be discriminated among human tissues. Conclusions Our results suggest that although cells having NCSC-like properties present the same phenotype, they differ in the expression of certain markers and differentiation abilities. The present study is the first to demonstrate the differences in differentiation ability and molecular markers among multilineage human APDCs, PDLDCs, and OMSDCs obtained from the same patients and, concomitantly, the same sites and identify these tissue-specific markers in the human tooth developmental process.

Microstructured Hyaluronic Acid Hydrogel for Tooth Germ Bioengineering

Tooth loss has been found to adversely affect not just masticatory and speech functions, but also psychological health and quality of life. Currently, teeth replacement options include dentures, bridges, and implants. However, these artificial replacement options remain inferior to biological replacements due to their reduced efficiency, the need for replacements, and the risk of immunological rejection. To this end, there has been a heightened interest in the bioengineering of teeth in recent years. While there have been reports of successfully regenerated teeth, controlling the size and shape of bioengineered teeth remains a challenge. In this study, methacrylated hyaluronic acid (MeHA) was synthesized and microstructured in a hydrogel microwell array using soft lithography. The resulting MeHA hydrogel microwell scaffold resembles the shape of a naturally developing human tooth germ. To facilitate the epithelial–mesenchymal interactions, human adult low calcium high temperature (HaCaT) cells were seeded on the surface of the hydrogels and dental pulp stem cells (DPSCs) were encapsulated inside the hydrogels. It was found that hydrogel scaffolds were able to preserve the viability of both types of cells and they appeared to favor signaling between epithelial and mesenchymal cells, which is necessary in the promotion of cell proliferation. As such, the hydrogel scaffolds offer a promising system for the bioengineering of human tooth germs in vitro.

Effect of Strontium-Doped Bioactive Glass on Preventing Formation of Demineralized Lesion

This study investigated the effect of strontium-doped bioactive glass (SBAG) on the formation of dental demineralized lesions. Materials and methods: The study materials were 48 sound human tooth specimens with both dentine and enamel, divided equally into four groups: Group 1 (SBAG), Group 2 (SBAG+Fluoride), Group 3 (Fluoride), and Group 4 (Water as control). After 14 days of pH cycling, the surface morphology of the specimens was observed by scanning electron microscopy. Crystal characteristics of the precipitates were assessed by X-ray diffraction (XRD). Micro-CT was used to measure the mineral loss and the depths of the demineralized lesions formed. Results: Exposure of collagen in inter-tubular areas in dentine was seen in the control group (Group 4) but not in Groups 1 to 3. In Group 2, there were obvious granular particles on the surface of the dentine. XRD revealed precipitation of apatites on the surface of the tooth specimens in Groups 1 to 3. The mean lesion depths in dentine were 81.80 μm, 30.68 μm, 39.04 μm, and 146.36 μm in Groups 1 to 4, respectively (p < 0.001). Lesions in enamel were only found in the control group. The mean mineral loss values in the dentine lesions were 1.25 g/cm3, 0.88 g/cm3, 0.87 g/cm3, and 1.65 g/cm3, in Groups 1 to 4, respectively (p < 0.001). Conclusion: Strontium-doped bioactive glass has a preventive effect on the formation of demineralized lesions in enamel and dentine.

High Resolution STEM Images of the Human Tooth Enamel Crystals

High-resolution scanning transmission electron microscopy (STEM) images of human tooth enamel crystals, mainly in the high-angle annular dark-field (STEM-HAADF) mode, are presented in this work along the [1000], [10-11]. and [1-210] directions. These images allow knowing some structural details at the nanometric level of the human tooth enamel crystals and of the central dark line (CDL) observed at their centers. The transmission electron microscopy (TEM) and high-resolution TEM (HRTEM) images of the CDL showed the Fresnel contrast. In the STEM bright-field (STEM-BF) and annular-dark-field (STEM-ADF) images, the CDL was observed as an unstrain hydroxyapatite (HAP)-like zone but surrounded by a strained zone. In the STEM-HAADF images, the CDL appeared with a weak contrast, and its contrasts’ thickness was registered between 3 and 8 Å. The arrangement obtained in the STEM-HAADF images by identifying the bright points with the Ca atoms produced the superposition of the HAP atomic sites, mainly along the [0001] direction. The findings provide further information on the structure details at the center of enamel crystals, which favors the anisotropic carious dissolution at the CDL.