Molecular mechanisms of tooth, maxilla and mandible development / 口腔医学

@#Understanding the pattern and molecular mechanisms of tooth, maxilla and mandible development is the prerequisite for studying their regeneration. Tooth development can be divided into three stages: bud-bell stage, tooth crown development stage and tooth root development stage. During these processes, key genes show spatial and temporal expression pattern. Tooth development is a complex process involving interactions between dental epithelium and mesenchyme, precise regulations of enamel knots in cusp patterning, as well as successful eruption into the oral cavity under proper biomechanical stress and signaling transductions. The development of tooth, maxilla and mandible, all of which originate from the first branchial arch, is independent and regulates each other to form a whole during development. Any developmental defects of them will ultimately cause defects to the others. In this paper, we briefly reviewed the development of tooth, maxilla and mandible, proposed that the homeostasis of microenvironment is critical for their development. Moreover, we reviewed the role of Meckel’s cartilage, a special structure and signaling mechanism during mandible development. At last, we proposed an integrated development model of tooth, maxilla and mandible. We also hope that the regeneration of fully functional tooth, maxilla and mandible in human can be achieved based on fundamental knowledge we have gained so far.

Validity of Nolla’s Method for Age Estimation for Children in Kurdish Iraqi Population - A Retrospective Study

Context: Radiographs have an essential role in Chronological Age (CA) estimation and are being used for dental age (DA) determination. Aims: Detecting the validity of Nolla’s method (NM) for the age assessment of Kurdish Iraqi children (KIC). Methods and Material: A retrospective study was performed using orthopantomographs (OPGs) of 354 subjects aged from 4 to 13 years (178 boys and 176 girls) and their recording files. Subjects were divided into nine study groups: 4, 5, 6, 7, 8, 9, 10, 11, 12, and 13 years old. The chronological age (CA) was subtracted from the DA to find the validity of NM; the positive results indicated the overestimation of age, whereas the negative results indicated for underestimation. The data were recorded through a digitalised system using Microsoft Excel worksheet and analysed by Statistical Package for the Social Sciences (SPSS, version 25) programme using the dependent T?test and graphical analysis. The level of P value used in this study was set at < 0.05. Results: The DA is underestimated in ages 9 to 13 in boys and girls. The highest difference in DA–CA was at the age of 9 years (?0.146 ± 0.162). Conclusions: NM for age estimation was slightly overestimated in age groups of 4, 5, 6, 7, and 8 years in boys and girls without statistically significant differences. However, this method underestimated the ages of KIC ranging from 9 to 13 years significantly.

Research progress on the role of transforming growth factor-β in tooth and craniofacial bone development and diseases / 口腔疾病防治

@#Dental and craniofacial bone development is a highly coordinated process that is tightly controlled by genetics and influenced by complex environments. The abnormal regulation of many development-related signaling molecules may lead to abnormal tooth development, severe craniofacial bone formation disorders, and developmental deformities. Transforming growth factor-β (TGF-β) is widely expressed in vivo and participates in many cellular biological processes, showing complex regulatory roles in mammalian craniofacial bone growth and tooth development. In tooth development, abnormal TGF-β signaling can lead to the failure of tooth germ formation, and its deletion mutation can directly affect odontoblast differentiation and enamel formation defects. However, the current research on TGF-β mainly focuses on the early stage of tooth development, and a comprehensive and systematic study of TGF-β-related tooth development is lacking. TGF-β signal transduction mainly controls the development of teeth and craniofacial bone by regulating the expression of development-related molecules via the classical Smad-dependent signaling pathway. In addition, the nonclassical mitogen-activated protein kinase (MAPK) pathway also participates in this process. Abnormal TGF-β signaling may cause jaw development disorders, temporomandibular joint dysplasia and inflammation, and cleft palate. Because the specific regulatory mechanism of TGF-β in craniofacial bone development has not been fully elucidated, its specific application in the treatment of related diseases is also greatly limited. This paper describes the new research progress of TGF-β in the development of teeth, jaws, temporomandibular joints and palate as well as related diseases.

Temporo-spatial distribution of stem cell markers CD146 and p75NTR during odontogenesis in mice

Abstract Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development. Objective Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis. Methodology The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry. Results Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR. Conclusions These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.

Transcriptome sequencing technology and its application in oral diseases, dental implants and regeneration / 中国组织工程研究

BACKGROUND: Since the application of transcriptome sequencing technology has made remarkable achievements in the research of melanoma and breast cancer, transcriptome sequencing technology has become a hot research method in scientific research and widely used in the field of stomatology. OBJECTIVE: To review the development of transcriptome sequencing technology and its application in various disciplines of stomatology through searching, screening and reading literature. METHODS: A search of CNKI, CBM and PubMed was performed for relevant literature published from 2015 through 2020. The search terms were “transcriptome, sequencing technology, RNA-seq, microarray, oral cancers, OSCC, periodontal, caries, pulp disease, tooth development, DPSCs, PDLSCs, orthodontics, implant” in English and Chinese, respectively. According to the inclusion and exclusion criteria, 72 literatures were included to review the development of transcriptome sequencing technology and its application in the field of stomatology. RESULTS AND CONCLUSION: In the development of transcriptome sequencing technology, RNA-seq technology has the greatest advantage in scientific research, because of its high accuracy, high throughput and low price. In the field of stomatology, this technology has been used in the research of oral squamous cell carcinoma, periodontitis, dental pulp disease, tooth regeneration, orthodontics and dental implantation, and has achieved some achievements. However, the current research on the same kind of disease does not reflect the relationship between the various studies, and the research content is limited. It is believed that more discoveries can be yielded in stomatology by exploring the relationship between different studies and expanding the research content.

Sp7/osterix positively regulates dlx2b and bglap to affect tooth development and bone mineralization in zebrafish larvae

Osterix (or Sp7) is an important transcription factor that promotes osteoblast differentiation by modulating the expression ofa range of target genes. Although many studies have focused on Osterix/Sp7 regulatory mechanisms, the detailed functionshave not been fully elucidated. Toward this end, in this study, we used CRISPR/Cas9 technology to knock out the zebrafishsp7 gene, and then analyzed its phenotype and biological function. Two knockout sp7 mutant lines were successfullyobtained. The bone mineralization level was significantly reduced in the zebrafish sp7-/- homozygote, resulting inabnormal tooth development in the larvae. Quantitative real-time polymerase chain reaction showed that loss of sp7 led todown-regulated expression of the dlx2b and bglap genes related to tooth development and bone mineralization, respectively. Moreover, cell transfection experiments demonstrated that Sp7 directly regulates the expression of dlx2b and bglapthrough Sp7-binding sites on the promoter regions of these two genes. Overall, this study provides new insight into the roleof Sp7 in bone mineralization and tooth development.

Development of Mandibular Third Molars in 18 Year Old North Indian Population

The aim of this study was to investigate development of third molars in 18 year old North Indian male population by means of CT scans and 3D software imaging. Methods: A retrospective analysis of CT scans of 50 patients aged 18 years at the time of their scan was conducted, and the developmental stages of the left third molars were evaluated using Demirjian's classification. Demirjian's classification system distinguished eight stages of crown and root development (Stages A-H). Results: The percentile distributions were recorded for each stage of development and variations for different stages were noted. The developmental and physiological changes of the tooth can be correlated to chronological age. Conclusion: The present investigation could provide reference data for third molar development in the North Indian population and maybe used in wider studies of tooth development and age prediction using dental radiographs for forensic and medico- legal purposes.

Expression patterns of ectodysplasin and ectodysplasin receptor during early dental development in zebrafish / 华西口腔医学杂志

OBJECTIVE@#This study aims to study the expression patterns of ectodysplasin (EDA) and ectodysplasin receptor (EDAR) during the early development of zebrafish and provide a foundation for further research of the Eda signaling pathway in tooth development.@*METHODS@#Total RNA was extracted from zebrafish embryos at 48 hours postfertilization (hpf) and then reverse transcribed for cDNA library generation. The corresponding RNA polymerase was selected for the synthesis of the digoxin-labeled antisense mRNA probe of zebrafish pharyngeal tooth specific marker dlx2b and Eda signaling-associated genes eda and edar in vitro. The three sequences were ligated into a pGEMT vector with a TA cloning kit, and polymerase chain reaction (PCR) was applied to linearize the plasmid. The resultant PCR sequences were used as templates for synthesizing Dig-labeled mRNA probe dlx2b, eda, and edar. Zebrafish embryos were collected at 36, 48, 56, 60, 72, and 84 hpf, then whole mount in situ hybridization was performed for the detection of eda and edar expression patterns. Then, their expression patterns at 72 hpf were compared with the expression pattern of dlx2b.@*RESULTS@#The mRNA antisense probes of dlx2b, eda, and edar were successfully obtained. The positive signals of eda and edar were observed in zebrafish pharyngeal tooth region at 48-72 hpf and thus conform to the signals of dlx2b in the positive regions.@*CONCLUSIONS@#The ligand eda and edar, which are associated with the Eda signaling pathway, are strongly expressed only at the pharyngeal tooth region in zebrafish from tooth initiation to the morphogenesis stage. Thus, the Eda signaling pathway may be involved in the regulation of the early development of zebrafish pharyngeal teeth.

Expression and Localization of Keap1 During Amelogenesis in the Developing Molar Germ of Rats

The objective of this study was to examine the expression pattern of Kelch-like ECH-associated protein 1 (Keap1) in the maxillary 2(nd) molar germs of rats. We used the maxillary 2(nd) molar germs in rats' pup at postnatal day 3 (bell stage), 6 (crown formation stage) and 9 (root formation stage). The investigation on mRNA and protein levels were done using reverse transcription - polymerase chain reaction and western blot. Localization of Keap 1 in the maxillary 2(nd) molar germs were revealed through immunofluorescence staining. Keap1 from the maxillary 2nd molar germs were mostly manifested on postnatal day 3 and dramatically decreased on postnatal day 6 and 9 at mRNA and protein levels, while amelogenin and ameloblastin increased during the development of maxillary 2nd molar germs. During immunofluorescence analysis, the strong immunoreactivity against Keap1 was detected in the apical side of ameloblasts at the presecretory and secretory stages. However, Keap1 expression was hardly observed in the ameloblasts at the maturation stage. These results shows that Keap1 is strongly expressed in the presecretory and secretory ameloblasts of amelogenesis, and suggest that Keap1 may be a crucial molecule for the regulatory mechanisms tasked with the formation of enamel layer.

Advances in the p75 neurotrophic factor receptor in stomatology / 中华口腔医学杂志

The p75 neurotrophic factor receptor is a low affinity receptor for neurotrophic factors and plays an important role in nerve growth, development and function integrity. It is closely related to dental development, oral and maxillofacial tumor, nerve repair and tissue engineering. It shows good prospect for application. In this paper, the research progress of p75 neurotrophic factor receptor in Stomatology is reviewed.

Tooth Agenesis and Delay in Patients with Agenesis of Mandibular Second Premolars / 대한소아치과학회지

Tooth agenesis is a common human dental anomaly and the agenesis of mandibular second premolars has been proven to be the most frequently observed. The aim of this study is to investigate tooth agenesis and delayed tooth development in patients with agenesis of mandibular second premolars.This study reviewed 9 to 15 year-old patients with agenesis of mandibular second premolars who visited the department of pediatric dentistry of Yonsei University Dental Hospital and took panoramic radiographs from January 2014 to December 2016. On panoramic radiographs, agenesis of teeth was observed and developmental delay of teeth was evaluated by the Nolla method. Among 125 patients with agenesis of mandibular second premolars, 58 patients (46.4%) showed agenesis of other teeth and 38 patients (30.4%) showed delayed tooth development. In this study, patients with agenesis of mandibular second premolars were more likely to have tooth agenesis or delayed eruption of other teeth.

Expression characteristics of Wnt5a in different periods of cementum development and its effect on differentiation of cementoblasts of mice / 吉林大学学报(医学版)

Objective: To study the expression characteristics of Wnt5a in cementum of the mice after birth and the effect of Wnt5a on the differentiation of cementoblasts in vitro, and to illuminate the mechanism of Wnt noncanonical signaling in regulating the tooth development. Methods: The postnatal Kunming mice at the days 0. 5, 4. 5, 12. 5, 18. 5, 24. 5 and 30. 5 after birth were selected and divided into various groups by different time points (n=3). The mice were sacrificed and the lower first molar and peridentium were prepared. Immunohistochemical staining was used to detect the expression characteristics of Wnt5a in tooth and peridentium of the mice. The OCCM30 cells were divided into control group and experiment group. 200 μg · L-1 recombinant Wnt5a protein were added in experiment group, and nothing was added in control group; the total RNA in the cells was exrracted. The relative expression levels of Ocn, Opn, Alp, Bsp, Wnt5a, and Runx-2 genes in the OCCM30 cells in two groups were detected by RT-PCR method. Results: At the day 0. 5 after birth, the expression of Wnt5a in the lower first molar tooth germ was negative. At the day 4. 5 after birth, the expression of Wnt5a in ameloblasts and odontoblasts were positive. At the day 16. 5 after birth, the expression of Wnt5a in cementoblasts was positive. At the days 20.5 - 30.5 after birth, the expressions of Wnt5a in odontoblasts and periodontal ligament cells were positive. Compared with control group, the relative expression levels of Ocn and Alp gene in the OCCM30 cells in experiment group were decreased (P<0. 05). Conclusion: The expression of Wnt5a is regular at different tooth developmental stages after birth. Wnt5a can reduce the expression of differentiated genes and play an important role in the growth of cementum.

State of the art of bulk-fill resin-based composites: a review / Revisión del estado actual de resinas compuestas bulk-fill

ABSTRACT. Introduction: ameloblasts are cells responsible for the production and mineralization of the organic matrix of enamel through several stages: pre-secretory, secretory, transition, and maturation. The organic matrix components are produced in the secretory phase. In the maturation phase, the organic component is removed and the mineralization process starts. This process requires the involvement of matrix metalloproteinase 20 (MMP-20), also called enamelysin. Several studies have shown the presence of MMP-20 in tooth development and its relationship to alterations in enamel formation. The objective was: to classify the different studies and laboratory techniques used to demonstrate the involvement of enamelysin in tooth development and its relation to pathologies during enamel formation. Methods: a systematic review was conducted with the following bibliographic databases: PubMed, Science-Direct, Hinari, and SciELO, in order to classify the different studies related to the involvement of MMP-20 in tooth development and the methods to detect its expression, between the years of 2009 and 2014. Results and conclusions: 11 in vitro models show that MMP-20 has specific cleavage sites for enamel matrix proteins. This process is altered by chemical composition, ions, and the presence of hydroxyapatite. Enamel morphology is altered in the knockout models. In human studies, MMP-20 has been associated with increased susceptibility to dental caries, enamel thickness, and dental agenesis.

RESUMEN. Introducción: el ameloblasto es la célula encargada de la producción y mineralización de la matriz orgánica del esmalte. Atraviesa varias etapas: la fase pre-secretora, secretora, de transición y maduración. En la fase secretora se producen los componentes de la matriz orgánica. En la fase de maduración se elimina el componente orgánico y se inicia el proceso de mineralización. Este proceso requiere de la participación de la metaloproteinasa de matriz 20 (MMP-20) o también llamada enamelisina. Diversos estudios demuestran la presencia de MMP-20 en el desarrollo dentario y su relación con alteraciones en la formación del esmalte. El objeto fue clasificar los diferentes estudios y técnicas de laboratorio empleadas que demuestren la participación de enamelisina en el desarrollo dentario y su relación con patologías en la formación del esmalte. Métodos: se realizó una revisión sistemática de la literatura con las siguientes bases bibliográficas: PubMed, Science-Direct, Hinari y SciELO, con el fin de clasificar los diferentes estudios relacionados con la participación de MMP-20 en el desarrollo dental y los métodos utilizados para detectar su expresión, entre los años de 2009 a 2014. Resultados y conclusiones: los modelos in vitro evidencian que MMP-20 tiene sitios específicos de escisión para las proteínas de matriz de esmalte. Este proceso se ve alterado por la composición química, iones, y la presencia de hidroxiapatita. En los modelos knockout la morfología del esmalte está alterada. En los estudios en humanos, se ha relacionado la MMP-20 con una mayor susceptibilidad de caries dental, el grosor completo de esmalte y agenesias dentales.

The role of enamelysin (MMP-20) in tooth development: systematic review / El papel de la enamelisina (MMP-20) en el desarrollo dentario: revisión sistemática

ABSTRACT. Introduction: ameloblasts are cells responsible for the production and mineralization of the organic matrix of enamel through several stages: pre-secretory, secretory, transition, and maturation. The organic matrix components are produced in the secretory phase. In the maturation phase, the organic component is removed and the mineralization process starts. This process requires the involvement of matrix metalloproteinase 20 (MMP-20), also called enamelysin. Several studies have shown the presence of MMP-20 in tooth development and its relationship to alterations in enamel formation. The objective was: to classify the different studies and laboratory techniques used to demonstrate the involvement of enamelysin in tooth development and its relation to pathologies during enamel formation. Methods: a systematic review was conducted with the following bibliographic databases: PubMed, Science-Direct, Hinari, and SciELO, in order to classify the different studies related to the involvement of MMP-20 in tooth development and the methods to detect its expression, between the years of 2009 and 2014. Results and conclusions: 11 in vitro models show that MMP-20 has specific cleavage sites for enamel matrix proteins. This process is altered by chemical composition, ions, and the presence of hydroxyapatite. Enamel morphology is altered in the knockout models. In human studies, MMP-20 has been associated with increased susceptibility to dental caries, enamel thickness, and dental agenesis.

RESUMEN. Introducción: el ameloblasto es la célula encargada de la producción y mineralización de la matriz orgánica del esmalte. Atraviesa varias etapas: la fase pre-secretora, secretora, de transición y maduración. En la fase secretora se producen los componentes de la matriz orgánica. En la fase de maduración se elimina el componente orgánico y se inicia el proceso de mineralización. Este proceso requiere de la participación de la metaloproteinasa de matriz 20 (MMP-20) o también llamada enamelisina. Diversos estudios demuestran la presencia de MMP-20 en el desarrollo dentario y su relación con alteraciones en la formación del esmalte. El objeto fue clasificar los diferentes estudios y técnicas de laboratorio empleadas que demuestren la participación de enamelisina en el desarrollo dentario y su relación con patologías en la formación del esmalte. Métodos: se realizó una revisión sistemática de la literatura con las siguientes bases bibliográficas: PubMed, Science-Direct, Hinari y SciELO, con el fin de clasificar los diferentes estudios relacionados con la participación de MMP-20 en el desarrollo dental y los métodos utilizados para detectar su expresión, entre los años de 2009 a 2014. Resultados y conclusiones: los modelos in vitro evidencian que MMP-20 tiene sitios específicos de escisión para las proteínas de matriz de esmalte. Este proceso se ve alterado por la composición química, iones, y la presencia de hidroxiapatita. En los modelos knockout la morfología del esmalte está alterada. En los estudios en humanos, se ha relacionado la MMP-20 con una mayor susceptibilidad de caries dental, el grosor completo de esmalte y agenesias dentales.

Osteocalcin Expression and Mineralization in Developing Tooth of Xenopus laevis

Osteocalcin (OC) is the most abundant noncollagenous protein of extracellular matrix in the bone. In an OC deficient mouse, bone formation rates are increased in cancellous and cortical bones. OC is known as a negative regulator of mineral apposition. OC is also expressed in the tooth of the rat, bovine, and human. However, little is known about OC during tooth development in Xenopus. The purpose of this study is to compare the expression of OC with mineralization in the developing tooth of Xenopus, by using von Kossa staining and in situ hybridization. At stage 56, the developmental stage of tooth germ corresponds to the cap stage, and an acellular zone was apparent between the dental papilla and the enamel organ. From stage 57, calcium deposition was revealed by von Kossa staining prior to OC expression, and the differentiated odontoblasts forming predentin were located at adjoining predentin. At stage 58, OC transcripts were detected in the differentiated odontoblasts. At stage 66, OC mRNA was expressed in the odontoblasts, which was aligned in a single layer at the periphery of the pulp. These findings suggest that OC may play a role in mineralization and odontogenesis of tooth development in Xenopus.

Smad4 Mediated TGF-beta/BMP Signaling in Tooth Formation Using Smad4 Conditional Knockout Mouse

Expression Patterns of Ki-67, Cyclin A, and Cyclin D1 during Tooth Development / 체질인류학회지

Dental epithelial and mesenchymal cells that form the teeth undergo dynamic changes in cell cycle during tooth development and morphogenesis. Although proliferation has been known as a key event during odontogenesis, the cell cycle phases and their relations with the complicated molecular mechanisms of tooth development are not fully understood yet. This study comparatively examined the expression patterns of Ki-67, cyclin A, and cyclin D1 during tooth development in the mouse incisor and molar in order to identify the cell-cycle characteristics during odontogenesis. We found that Ki-67 and cyclin A were expressed in the proliferating cells in the dental epithelial and mesenchymal tissues at the bud, cap and bell stages. Cycln D1 showed distinct expression in the incisor odontoblast region and the enamel knot, in which Ki-67 nor cyclin A was expressed. Our results provide specific information on the cell cycle phases during tooth development that may provide clues to relate them with the complex odontogenic mechanisms. Furthermore, we suggest that our findings enlightened the previous studies on the incisor odontoblasts and the enamel knot during tooth development.

Immunolocalization of FGF-10 in murine incisors and molar germs / Imunolocalização de FGF-10 em germes de incisivos e molares murinos

Objective: The aim of this study was to identify the presence of FGF-10 in mouse dental germs by means of the immunohistochemical technique, fromthe initial development phase through to the more advanced phases. Methods: Fetuses of five mice, on days 15.5, 16.5, 17.5, 18.5 and 19.5 of pregnancy, respectively, were collected. At time intervals of 0.5, 1.5, 2.5 and 3.5 days after birth, the mouse offspring were sacrificed. The heads of all the specimens were fixed and submitted to histotechnique and3?m thick sections were obtained. The presence of FGF-10 was detected by means of the avidin-biotin-peroxidase immunohistochemistry technique.Results: Immunostaining was detected in both epithelium and ectomesenchyme with intensity and spatial-temporal differences. A reduction in the presence of FGF-10 was observed in the cervical loop area, on the lingual side of the incisor crowns, and both sides of the molar crowns. With the increase in enamel matrix deposition, immunostaining on the secretory pole of ameloblasts also increased. Conclusion: FGF-10 immunostaining could be related to cell proliferation in epithelium and cell differentiation in epithelium and ectomesenchyme. This could be related to morphological determination in intercuspal areas of molar germs and to continuous growth of the incisor crown. Decrease in FGF-10 in the cervical loop could be related to the termination of crown formation. Increase in FGF-10 expression in ameloblasts suggests a relationship with active enamel production. The results suggest the inclusion of the pattern of the presence of FGF-10 in future investigations into the cause of morphological anomalies, such as palato-gingival groove.

Objetivo: Identificar a presença de FGF-10 em germes dentários de rato pela técnica de imunohistoquímica. Métodos: Os fetos de cinco ratos, nos dias 15,5; 16,5; 17,5; 18,5; 19,5, respectivamente, de gravidez, foram coletados. Em intervalos de tempo de 0,5; 1,5; 2,5 e 3,5 dias após o nascimento, as proles de ratas foram sacrificadas. As cabeças de todos os exemplares foram fixadas e submetidas à histotécnica e cortes com 3?m de espessura foram obtidos. A presença de FGF-10 foi detectada pela técnica de imunohistoquímica da avidinabiotina-peroxidase. Resultados: A imunomarcação foi detectada no epitélio e no ectomesênquima com intensidade e diferenças espaço-temporais. Uma redução da presença de FGF-10 foi observada na área da alça cervical, no lado lingual de coroas de incisivo e em ambos os lados das coroas de molares. Com o aumento da deposição de matriz de esmalte, a imunomarcação no pólo secretor de ameloblastos também aumentou.Conclusão: a presença de FGF-10 parece estar relacionada com a proliferação de células no epitélio e diferenciação de células no epitélio e ectomesênquima. Isso pode estar relacionado à determinação morfológica nas áreas intercuspídeas de germes dos molares e ao crescimento contínuo da coroa nos incisivos. A diminuição do FGF-10 em áreas alça cervical parece estar relacionada com o término de formação da coroa. O aumento de FGF-10 em ameloblastos parece estar relacionada com a produção ativa de esmalte.

Differential Expression of Osteonectin in the Rat Developing Molars

Tooth development involves bud, cap, bell and hard tissue formation stages, each of which is tightly controlled by regulatory molecules. The aim of this study was to identify genes that are differentially expressed during dental hard tissue differentiation. Sprague-Dawley rats at postnatal days 3, 6 and 9 were used in the analysis. Differential display RT-PCR (DD-PCR) was used to screen differentially expressed genes between the 2nd (root formation stage, during mineralization) and 3rd (cap stage, before mineralization) molar germs at postnatal day 9. The DNA detected in the 2nd molar germs showed homology to osteonectin only (GenBank accession no. NM_012656.1). The level of osteonectin mRNA expression was much higher in the 2nd molar germs than in the 3rd molar germs and was found to increase in a time-dependent manner from the early bell stage to the root formation stage in the 2nd molar germs. The pattern of osteonectin protein expression was consistent with these RT-PCR results. Osteonectin protein was found by immunofluorescent analysis to localize in odontoblasts and preodontoblasts rather than the dentin matrix itself. Further studies are needed to validate the involvement of osteonectin in mineralization and root formation.

Expression of Tissue Inhibitors of Metalloproteinases in Developing Rat Tooth Germs / 체질인류학회지

Tissue inhibitors of metalloproteinases (TIMPs) are a family of secreted molecules that were identified as natural inhibitors of matrix metalloproteinases (MMPs). Tooth histomorphogenesis and cytodifferentiation are accompanied by rapid changes in cellular organization and remodeling of the extracellular matrix, in which MMPs and TIMPs might be expected to play significant roles. This study examined the expression and localization of TIMP-1 and TIMP-2 during the molar development of rats. The expression patterns of TIMPs were determined from Sprague-Dawley rat pups including the developing molars using RT-PCR, western blot and immunofluorescent staining. Gene and protein quantification analyses showed that both TIMPs increased from the cap stage to the root stage tooth germs. In contrast, the immunofluorescent data showed that they were expressed slight differentially. TIMP-1 was strongly expressed in secretory ameloblasts and moderate immunoreactivity was observed along the basement membrane. TIMP-2 expression was also detected in the basement membrane. Although strong immunoreactivity was observed in the secretory ameloblasts and enamel matrix itself, differentiated odontoblasts showed weak reactivity. However, little reactivity for both TIMPs were detected in the cap stage tooth germs and surrounding tissues. These distinct temporospatial expression patterns of TIMPs suggest that the TIMPs may play a variety of roles including dental hard tissue formation during molar tooth development.