mTORC1 signaling pathway regulates tooth repair / 国际口腔科学杂志·英文版

Tooth germ injury can lead to abnormal tooth development and even tooth loss, affecting various aspects of the stomatognathic system including form, function, and appearance. However, the research about tooth germ injury model on cellular and molecule mechanism of tooth germ repair is still very limited. Therefore, it is of great importance for the prevention and treatment of tooth germ injury to study the important mechanism of tooth germ repair by a tooth germ injury model. Here, we constructed a Tg(dlx2b:Dendra2-NTR) transgenic line that labeled tooth germ specifically. Taking advantage of the NTR/Mtz system, the dlx2b+ tooth germ cells were depleted by Mtz effectively. The process of tooth germ repair was evaluated by antibody staining, in situ hybridization, EdU staining and alizarin red staining. The severely injured tooth germ was repaired in several days after Mtz treatment was stopped. In the early stage of tooth germ repair, the expression of phosphorylated 4E-BP1 was increased, indicating that mTORC1 is activated. Inhibition of mTORC1 signaling in vitro or knockdown of mTORC1 signaling in vivo could inhibit the repair of injured tooth germ. Normally, mouse incisors were repaired after damage, but inhibition/promotion of mTORC1 signaling inhibited/promoted this repair progress. Overall, we are the first to construct a stable and repeatable repair model of severe tooth germ injury, and our results reveal that mTORC1 signaling plays a crucial role during tooth germ repair, providing a potential target for clinical treatment of tooth germ injury.

Tooth number abnormality: from bench to bedside / 国际口腔科学杂志·英文版

Tooth number abnormality is one of the most common dental developmental diseases, which includes both tooth agenesis and supernumerary teeth. Tooth development is regulated by numerous developmental signals, such as the well-known Wnt, BMP, FGF, Shh and Eda pathways, which mediate the ongoing complex interactions between epithelium and mesenchyme. Abnormal expression of these crutial signalling during this process may eventually lead to the development of anomalies in tooth number; however, the underlying mechanisms remain elusive. In this review, we summarized the major process of tooth development, the latest progress of mechanism studies and newly reported clinical investigations of tooth number abnormality. In addition, potential treatment approaches for tooth number abnormality based on developmental biology are also discussed. This review not only provides a reference for the diagnosis and treatment of tooth number abnormality in clinical practice but also facilitates the translation of basic research to the clinical application.

Micrometric measurement of the density of stained odontoblast processes

The embryological, structural and functional unit of the dentine-pulp complex shares the odontoblast, located in the border of the dentine pulp, with basal nuclei and organelles. The odontoblast process emerges from its apical pole. It is formed by microtubules, microfilaments and vesicles covered by membranes penetrating the dentinal tubules, isolated from the inter-tubular matrix, along the extent of the dentine. The objective of this study was to evaluate the efficacy of three staining techniques: hematoxylin-eosin, periodic acid-Schiff and Schmorl, by staining the process, from beginning to end, and compare the results with the erosion technique. Thirty human teeth were employed in the trial; after their extraction the pulp was fixated, the pieces demineralized in nitric acid at 8%, the collagen filaments eliminated with Type II Collage-nase, the tissue was stained, and the measurements were made. The portions with no pulp were prepared with the erosion technique. Results: Comparing the best results obtained by staining with the values obtained with the erosion technique, the former showed lower values. Conclusion: Staining techniques show lower density of the staining processes compared with the dentinal tubules in the erosion technique.

Dentin permeability of the apical third in different groups of teeth

This ex vivo study evaluated dentin permeability of the root canal in the apical third of different human groups of teeth. Eighty teeth were used, 8 from each dental group: maxillary and mandibular central incisors, lateral incisors and canines, maxillary first premolars (buccal and palatal roots), mandibular first premolars, and maxillary and mandibular second premolars, totalizing 88 roots that were distributed in 11 groups. The root canals were instrumented, irrigated with 1 percent NaOCl and 15 percent EDTA. Roots were immersed in 10 percent copper sulfate for 30 min and then in 1 percent rubeanic acid alcohol solution for the same period; this chemical reaction reveals dentin permeability by the formation of copper rubeanate, which is a dark-colored compound. Semi-serial 100-µm-thick cross-sections were obtained from the apical third of the roots. Five sections of each apical third were washed, dehydrated, cleared and mounted on glass slides for examination under optical microscopy. The percentage of copper ion infiltration and the amount of tubular dentin were quantified by morphometric analysis. The penetration of copper ions in the apical third ranged from 4.60 to 16.66 percent. The mandibular central and lateral incisors presented the highest dentin permeability (16.66 percent), while the maxillary canines and mandibular second and first premolars presented the lowest dentin permeability (4.60 percent, 4.80 percent and 5.71 percent, respectively; p<0.001). The other teeth presented intermediate permeability. In conclusion, dye penetration into dentin tubules at the apical region is strongly dependent on the group of teeth evaluated.

Este estudo ex vivo avaliou a permeabilidade da dentina do canal radicular do terço apical de diferentes grupos de dentes humanos. Foram utilizados 80 dentes, sendo 8 de cada grupo dental superior e inferior: incisivos centrais, incisivos laterais, caninos, primeiros pré-molares superiores (raízes vestibulares e palatinas), primeiros pré-molares inferiores, segundos pré-molares superiores e inferiores, totalizando 88 raízes, as quais foram distribuídas em 11 grupos. Os canais foram instrumentados, irrigados com NaOCl a 1 por cento e EDTA a 15 por cento. As raízes foram imersas em sulfato de cobre a 10 por cento por 30 min e acido rubeânico a 1 por cento pelo mesmo período. Esta reação química revela a permeabilidade da dentina por meio da formação de um complexo escurecido denominado rubeanato de cobre. Hemi-secções de 100 µm de espessura foram obtidas do terço apical da raiz. Cinco secções do terço apical foram lavadas, desidratadas, diafanizadas e montadas em lâminas para análise em microscopia óptica. A porcentagem de infiltração de íons cobre e a quantidade de dentina tubular foram quantificadas por meio de análise morfométrica. A penetração de íons cobre no terço apical da raiz variou de 4,60 por cento a 16,66 por cento. Os incisivos centrais e laterais apresentaram a maior permeabilidade dentinária (16,66 por cento), e os caninos superiores e segundos e primeiros pré-molares inferiores as menores (4,60 por cento, 4,80 por cento e 5,71 por cento, respectivamente; p<0,001). Os outros dentes apresentaram permeabilidade intermediaria. Conclui-se que a penetração de corante nos túbulos dentinários da região apical é extremamente dependente do grupo de dentes avaliado.

Homologous amelogenin gene test of archaeological samples / 法医学杂志

OBJECTIVE@#Based on the sequence differences of Amelogenin homologous gene in the X and Y chromosomes, a pair of specific primers was designed to identify the sex of archaeological samples.@*METHODS@#Ancient DNA fragments were extracted from the bones and teeth of sacrificial slaves with an improved method that combines phenol-chloroform extraction, silicon dioxide adsorption with ultrafiltration concentration. The polyacrylamide gel electrophoresis (PAGE) was used to detect PCR products.@*RESULTS@#Seven in sixteen samples from eight graves showed positive results and the targeted segments were visible: a male with two bands of 106bp (Amel-X) and 112 bp (Amel-Y), while a female with only one band of 106 bp (Amel-X). Ancient DNA analyzing results from tooth samples are more marked than that from bones.@*CONCLUSION@#The improved extraction method is more effective for ancient DNA extraction, which reduced the PCR inhibitors and lowered experimental costs. The sex determination technology based on Amelogenin homologous gene is an important and feasible method in the molecular archaeological research.

Estudos macro e microscópico da morfologia do terço dentário apical na hipercementose: implicaçöes clínicas e etiopatogênicas / Macro and microscopical studies of apical dental portion morphology in the hypercementosis: clinical and etiopathogenic implications

Cento e sessenta dentes com e sem hipercementose foram analisados macroscopicamente, ao estereomicroscópio sem e com diafanizaçäo e na microscopia óptica de luz, com a finalidade de estudar a anatomia externa e interna do terço apical. Verificou-se que: 1) A hipercementose aumenta consideravelmente o número de canais secundários, canais acessórios e deltas apicais. 2) As modificaçöes na morfologia interna do terço apical dos dentes comprometidos näo säo passíveis de visualizaçäo e identificaçäo radiográfica. 3) O canal principal na grande maioria dos dentes com hipercementose apresenta-se constricto no terço apical associado à mudança da trajetória original ou a canais secundários, acessórios e deltas apicais. 4) O tipo morfológico mais comum de hipercementose dá à raiz a forma de clava. A forma localizada de hipercementose apresenta-se com considerável menor frequência. A forma de boca de manga de camisa encontra-se apenas eventualmente. 5) O padräo morfológico e aposicional do cemento constituinte da hipercementose assemelha-se ao cemento em dentes normais quando observado na microscopia óptica de luz, assim como sua superfície externa. Após estas verificaçöes, concluiu-se que na hipercementose o clínico de várias especialidades, como o cirurgiäo, o ortodontista e especialmente o endodontista, deve considerar a modificaçäo anatômica externa e interna a que foi submetido o dente portador, pois sua forma radicular tende a ser retentiva, o corpo radicular mais espesso e o terço apical mais rico em canalículos e deltas apicais associado à constricçäo e mudança direcional do canal principal