- Klíčová slova
- pulpální diagnostika,
- MeSH
- biologické markery MeSH
- diagnóza stomatologická metody MeSH
- dospělí MeSH
- lidé MeSH
- odontoblasty imunologie patologie MeSH
- pulpotomie metody MeSH
- zubní dřeň patologie MeSH
- zubní kaz * diagnóza imunologie prevence a kontrola MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Considerable amount of research has been focused on dentin mineralization, odontoblast differentiation, and their application in dental tissue engineering. However, very little is known about the differential role of functionally and spatially distinct types of dental epithelium during odontoblast development. Here we show morphological and functional differences in dentin located in the crown and roots of mouse molar and analogous parts of continuously growing incisors. Using a reporter (DSPP-cerulean/DMP1-cherry) mouse strain and mice with ectopic enamel (Spry2+/- ;Spry4-/- ), we show that the different microstructure of dentin is initiated in the very beginning of dentin matrix production and is maintained throughout the whole duration of dentin growth. This phenomenon is regulated by the different inductive role of the adjacent epithelium. Thus, based on the type of interacting epithelium, we introduce more generalized terms for two distinct types of dentins: cementum versus enamel-facing dentin. In the odontoblasts, which produce enamel-facing dentin, we identified uniquely expressed genes (Dkk1, Wisp1, and Sall1) that were either absent or downregulated in odontoblasts, which form cementum-facing dentin. This suggests the potential role of Wnt signalling on the dentin structure patterning. Finally, we show the distribution of calcium and magnesium composition in the two developmentally different types of dentins by utilizing spatial element composition analysis (LIBS). Therefore, variations in dentin inner structure and element composition are the outcome of different developmental history initiated from the very beginning of tooth development. Taken together, our results elucidate the different effects of dental epithelium, during crown and root formation on adjacent odontoblasts and the possible role of Wnt signalling which together results in formation of dentin of different quality. © 2021 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).
- MeSH
- buněčná diferenciace MeSH
- dentin * MeSH
- epitel MeSH
- extracelulární matrix - proteiny genetika MeSH
- myši MeSH
- odontoblasty * MeSH
- odontogeneze MeSH
- řezáky MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Understanding cell types and mechanisms of dental growth is essential for reconstruction and engineering of teeth. Therefore, we investigated cellular composition of growing and non-growing mouse and human teeth. As a result, we report an unappreciated cellular complexity of the continuously-growing mouse incisor, which suggests a coherent model of cell dynamics enabling unarrested growth. This model relies on spatially-restricted stem, progenitor and differentiated populations in the epithelial and mesenchymal compartments underlying the coordinated expansion of two major branches of pulpal cells and diverse epithelial subtypes. Further comparisons of human and mouse teeth yield both parallelisms and differences in tissue heterogeneity and highlight the specifics behind growing and non-growing modes. Despite being similar at a coarse level, mouse and human teeth reveal molecular differences and species-specific cell subtypes suggesting possible evolutionary divergence. Overall, here we provide an atlas of human and mouse teeth with a focus on growth and differentiation.
- MeSH
- buněčná diferenciace * genetika MeSH
- dospělí MeSH
- epitelové buňky MeSH
- genetická heterogenita MeSH
- kmenové buňky cytologie MeSH
- lidé MeSH
- mezoderm cytologie růst a vývoj metabolismus MeSH
- mladiství MeSH
- mladý dospělý MeSH
- modely u zvířat MeSH
- moláry cytologie růst a vývoj MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- odontoblasty MeSH
- řezáky cytologie růst a vývoj MeSH
- vývojová regulace genové exprese MeSH
- zuby cytologie růst a vývoj MeSH
- zvířata MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
To gain a better understanding of the progression of progenitor cells in the odontoblast lineage, we have examined and characterized the expression of a series of GFP reporters during odontoblast differentiation. However, previously reported GFP reporters (pOBCol2.3-GFP, pOBCol3.6-GFP, and DMP1-GFP), similar to the endogenous proteins, are also expressed by bone-forming cells, which made it difficult to delineate the two cell types in various in vivo and in vitro studies. To overcome these difficulties we generated DSPP-Cerulean/DMP1-Cherry transgenic mice using a bacterial recombination strategy with the mouse BAC clone RP24-258g7. We have analyzed the temporal and spatial expression of both transgenes in tooth and bone in vivo and in vitro. This transgenic animal enabled us to visualize the interactions between odontoblasts and surrounding tissues including dental pulp, ameloblasts and cementoblasts. Our studies showed that DMP1-Cherry, similar to Dmp1, was expressed in functional and fully differentiated odontoblasts as well as osteoblasts, osteocytes and cementoblasts. Expression of DSPP-Cerulean transgene was limited to functional and fully differentiated odontoblasts and correlated with the expression of Dspp. This transgenic animal can help in the identification and isolation of odontoblasts at later stages of differentiation and help in better understanding of developmental disorders in dentin and odontoblasts.
- MeSH
- buněčná diferenciace MeSH
- extracelulární matrix - proteiny genetika MeSH
- fluorescenční barviva MeSH
- fosfoproteiny genetika MeSH
- myši transgenní MeSH
- myši MeSH
- odontoblasty cytologie MeSH
- reportérové geny * MeSH
- sialoglykoproteiny genetika MeSH
- transgeny MeSH
- zelené fluorescenční proteiny genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, N.I.H., Extramural MeSH
In organized tissues, the precise geometry and the overall shape are critical for the specialized functions that the cells carry out. Odontoblasts are major matrix-producing cells of the tooth and have also been suggested to participate in sensory transmission. However, refined morphologic data on these important cells are limited, which hampers the analysis and understanding of their cellular functions. We took advantage of fluorescent color-coding genetic tracing to visualize and reconstruct in 3 dimensions single odontoblasts, pulp cells, and their assemblages. Our results show distinct structural features and compartments of odontoblasts at different stages of maturation, with regard to overall cellular shape, formation of the main process, orientation, and matrix deposition. We demonstrate previously unanticipated contacts between the processes of pulp cells and odontoblasts. All reported data are related to mouse incisor tooth. We also show that odontoblasts express TRPM5 and Piezo2 ion channels. Piezo2 is expressed ubiquitously, while TRPM5 is asymmetrically distributed with distinct localization to regions proximal to and within odontoblast processes.
- MeSH
- ameloblasty cytologie ultrastruktura MeSH
- buněčné jádro ultrastruktura MeSH
- buněčné výběžky ultrastruktura MeSH
- dentin ultrastruktura MeSH
- extracelulární matrix ultrastruktura MeSH
- fluorescenční protilátková technika MeSH
- iontové kanály ultrastruktura MeSH
- kationtové kanály TRPM ultrastruktura MeSH
- kompartmentace buňky MeSH
- mezenchymální kmenové buňky cytologie ultrastruktura MeSH
- mikroskopie elektronová rastrovací metody MeSH
- myši transgenní MeSH
- myši MeSH
- odontoblasty cytologie ultrastruktura MeSH
- řezáky cytologie ultrastruktura MeSH
- tvar buňky MeSH
- zobrazování trojrozměrné metody MeSH
- zubní dřeň cytologie ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Apoptosis during tooth development appears dependent on the apoptotic executioner caspase-3, but not caspase-7. Instead, activated caspase-7 has been found in differentiated odontoblasts and ameloblasts, where it does not correlate with apoptosis. To further investigate these findings, the mouse incisor was used as a model. Analysis of caspase-7-deficient mice revealed a significant thinner layer of hard tissue in the adult incisor. Micro computed tomography scan confirmed this decrease in mineralized tissues. These data strongly suggest that caspase-7 might be directly involved in functional cell differentiation and regulation of the mineralization of dental matrices.
- MeSH
- ameloblasty cytologie enzymologie metabolismus MeSH
- buněčná diferenciace * MeSH
- časové faktory MeSH
- imunohistochemie MeSH
- kaspasa 7 genetika metabolismus MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- odontoblasty cytologie enzymologie metabolismus MeSH
- odontogeneze MeSH
- proliferace buněk MeSH
- rentgenová mikrotomografie MeSH
- řezáky embryologie růst a vývoj metabolismus MeSH
- zubní sklovina embryologie růst a vývoj metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
OBJECTIVES: The primary enamel knot (PEK) is a population of cells that shows spatio-temporal restricted apoptosis during tooth development. It has been shown that caspase-9 and Apaf-1 are essential for apoptosis in the PEK as well as the central caspase-3. Caspase-7, as another executioner member in the caspase machinery, is considered to have caspase-3 like properties. DESIGN: The aim of this study was to detect caspase-7 activation during molar tooth development with a special focus on the cells of the PEK and to correlate the expression with the pattern of apoptosis and caspase-3 activation. Apoptosis in the PEK was investigated in caspase-7 deficient mice to examine the functional consequence of loss of this specific caspase. In addition, odontoblasts and ameloblasts, which are known to undergo cell death during their secretory and maturation stages, were investigated. RESULTS: Cleaved caspase-7 was found in the apoptotic region of the PEK, however, caspase-7-deficient mice still possessed apoptotic cells in the PEK in a similar distribution to the wild type. Caspase-7 is therefore not essential for apoptosis in the PEK. Notably, cleaved caspase-7-positive cells were found at later stages in odontoblasts and ameloblasts, but expression did not correlate with apoptosis in these tissues. CONCLUSIONS: The results indicate a non-essential apoptotic role of caspase-7 in the PEK apoptosis but suggest also possible non-apoptotic functions for caspase-7 in tooth development.
- MeSH
- ameloblasty cytologie MeSH
- apoptóza fyziologie MeSH
- kaspasa 7 nedostatek genetika metabolismus MeSH
- koncové značení zlomů DNA in situ MeSH
- moláry metabolismus MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- odontoblasty cytologie MeSH
- odontogeneze genetika fyziologie MeSH
- počítačová rentgenová tomografie MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The transcription factor c-Myb is involved in the control of cell proliferation, survival and differentiation. As these processes accompany the morphogenesis of developing teeth, this work investigates the possible role of c-Myb during odontogenesis. Analysis of the expression of c-Myb in the monophyodont mouse was followed by similar analysis in a diphyodont species, the pig, which has a dentition more closely resembling that of the human. The distribution of c-Myb was correlated with the pattern of proliferation and apoptosis and the tooth phenotype of c-Myb mutant mice was also assessed. In the mouse, c-Myb expression was detected throughout prenatal development of the first molar tooth. Negative temporospatial correlation was found between c-Myb expression and apoptosis, while c-Myb expression positively correlated with proliferation. c-Myb-positive cells, however, were more abundant than the proliferating cell nuclear antigen positive cells, suggesting other roles of c-Myb in odontogenesis. In the minipig, in contrast to the mouse, there was an asymmetrical arrangement of c-Myb positive cells, with a higher presence on the labial side of the tooth germ and dental lamina. A cluster of negative cells was situated in the mesenchyme close to the tooth bud. At later stages, the number of positive cells decreased and these cells were situated in the upper part of the dental papilla in the areas of future cusp formation. The expression of c-Myb in both species was strong in the odontoblasts and ameloblasts at the stage of dentin and enamel production suggesting a possible novel role of c-Myb during tooth mineralization.
- MeSH
- alely MeSH
- ameloblasty cytologie metabolismus MeSH
- apoptóza MeSH
- dentice MeSH
- druhová specificita MeSH
- embryo savčí cytologie embryologie metabolismus MeSH
- imunohistochemie MeSH
- klonování DNA MeSH
- koncové značení zlomů DNA in situ MeSH
- miniaturní prasata MeSH
- myši MeSH
- odontoblasty cytologie metabolismus MeSH
- odontogeneze MeSH
- osteoklasty cytologie metabolismus MeSH
- prasata MeSH
- proliferace buněk MeSH
- proliferační antigen buněčného jádra metabolismus MeSH
- proteiny buněčného cyklu genetika metabolismus MeSH
- protoonkogenní proteiny c-myb genetika metabolismus MeSH
- trans-aktivátory genetika metabolismus MeSH
- vývojová regulace genové exprese MeSH
- zuby cytologie embryologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The minipig provides an excellent experimental model for tooth morphogenesis because its diphyodont and heterodont dentition resemble that of humans. However, little information is available on the processes of tooth development in the pig. The purpose of this study was to classify the early stages of odontogenesis in minipigs from the initiation of deciduous dentition to the late bell stage when the successional dental lamina begins to develop. To analyze the initiation of teeth anlagens and the structural changes of dental lamina, a three-dimensional (3D) analysis was performed. At the earliest stage, 3D reconstruction revealed a continuous dental lamina along the length of the jaw. Later, the dental lamina exhibited remarkable differences in depth, and the interdental lamina was shorter. The dental lamina grew into the mesenchyme in the lingual direction, and its inclined growth was underlined by asymmetrical cell proliferation. After the primary tooth germ reached the late bell stage, the dental lamina began to disintegrate and fragmentize. Some cells disappeared during the process of lamina degradation, while others remained in small islands known as epithelial pearls. The minipig can therefore, inter alia, be used as a model organism to study the fate of epithelial pearls from their initiation to their contribution to pathological structures, primarily because of the clinical significance of these epithelial rests.
- MeSH
- bazální membrána embryologie MeSH
- buněčná diferenciace fyziologie MeSH
- dentin embryologie MeSH
- epitel embryologie MeSH
- mezoderm embryologie MeSH
- miniaturní prasata MeSH
- modely u zvířat MeSH
- morfogeneze fyziologie MeSH
- odontoblasty cytologie MeSH
- odontogeneze fyziologie MeSH
- orgán skloviny embryologie MeSH
- počítačové zpracování obrazu metody MeSH
- prasata MeSH
- premolár embryologie MeSH
- proliferace buněk MeSH
- proliferační antigen buněčného jádra analýza MeSH
- řezáky embryologie MeSH
- špičák embryologie MeSH
- zobrazování trojrozměrné metody MeSH
- zubní zárodek embryologie MeSH
- zuby mléčné embryologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
64 l. : il., tab. ; 30 cm
Stanovení fyzikálních charakteristik Nd:YAG laseru pro modifikaci povrchové morfologie exponovaného dentinu lidských extrahovaných molárů ve smyslu tubulové okluze. Vyhodnocení povrchové morfologie exponovaného dentinu ozářeného a neozářeného Nd:YAG laserem pomocí SEM (Scanning Electron Microscopy) a AFM (Atomic Force Microscopy). Zhodnocení přínosu AFM pro vyšetření morfologie exponovaného dentinu lidských extrahovaných zubů.; Determination of characteristics of Nd:YAG laser irradiation for the occluding of exposed dentinal tubules of extracted human molars. Characterisation of surface morphology of exposed dentin both irradiated and not irradiated by Nd:YAG laser by ScanningElectron Microscopy (SEM) and Atomic Force Microscopy (AFM). Determination of the usefulness of Atomic-force microscopy to the characterization of morphology of exposed dentin of extracted human teeth.
