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Radiological examination of the third molar is done in living individuals for estimation of chronological age, especially in the late adolescence. The aim of this study was to assess the application of Cameriere's third molar maturity index (I 3M ) to determine whether an individual is 18 years or older (adult) or younger than 18 years (minor) in a sample of Dutch individuals. The sample consisted of panoramic images of 360 individuals aged between 14 and 22 years old. Three observers performed the measurements. Gender was not statistically significant in discriminating adults and minors. The highest value of the Youden index of the receiver operating curve analysis was for the value of I 3M <0.08 in discriminating individuals as minor or adult. The specificity (Sp) and sensitivity (Se) results for females were 96.3% and 72.7% respectively. The Sp and Se for males were 95.0% and 84.0% respectively. The probabilities of correctly classified individuals were 83.3% and 88.9%, and Bayes post-test probability was 96.3% and 95.7% in females and males respectively. Obtained results showed that the specific cut-off point of I 3M <0.08 may be a useful and reliable method for adult age assessment in a Dutch population., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objectives: The aim of this prospective cohort study was to determine the effects of rapid maxillary expansion (RME) on the first molar roots of cleft lip and palate subjects along different root development stages., Materials and Methods: Thirty participants with unilateral cleft lip and palate were divided into three groups (n = 10), according to the type of expander used: Hyrax, iMini and Fan-type. A cone beam CT scan was performed before (T1) and 3 months after stabilization of the appliance (T2). Measurements of root lengths of the first permanent maxillary molars were taken to evaluate root development and external apical root resorption (EARR)., Results: Roots presenting open apexes before treatment exhibited a statistically significant increase in root length after treatment (P < .001). Conversely, there were no significant changes in the root length of roots that had closed apexes at the beginning of treatment (P > .05). Furthermore, there were no differences among groups, or correlation between cleft side and gender was found vis-à-vis to changes in the root length (p > .05)., Conclusions: Orthopaedic forces of RME were neither able to interrupt the root development process nor to cause EARR in cleft subjects., Clinical Relevance: As the RME is the most common orthodontic procedure in patients with cleft lip and palate, understanding its impact on dental structures is of fundamental importance.

This study provides regression equations for estimation of age of infants from the dimensions of their developing deciduous teeth. The sample comprises 97 individuals of known sex and age (62 boys, 35 girls), aged between 2 days and 1,081 days. The age-estimation equations were obtained for the sexes combined, as well as for each sex separately, thus including "sex" as an independent variable. The values of the correlations and determination coefficients obtained for each regression equation indicate good fits for most of the equations obtained. The "sex" factor was statistically significant when included as an independent variable in seven of the regression equations. However, the "sex" factor provided an advantage for age estimation in only three of the equations, compared to those that did not include "sex" as a factor. These data suggest that the ages of infants can be accurately estimated from measurements of their developing deciduous teeth., (© 2017 American Academy of Forensic Sciences.)

During tooth root development, stem cells from apical papillae (SCAPs) are indispensable, and their abilities of proliferation, migration and odontoblast differentiation are linked to root formation. Leucine-rich repeat-containing GPCR 4 (LGR4) modulates the biological processes of proliferation and differentiation in multiple stem cells. In this study, we showed that LGR4 is expressed in all odontoblast cell lineage cells and Hertwig's epithelial root sheath (HERS) during the mouse root formation in vivo. In vitro we determined that LGR4 is involved in the Wnt/β-catenin signaling pathway regulating proliferation and odonto/osteogenic differentiation of SCAPs. Quantitative reverse-transcription PCR (qRT-PCR) confirmed that LGR4 is expressed during odontogenic differentiation of SCAPs. CCK8 assays and in vitro scratch tests, together with cell cycle flow cytometric analysis, demonstrated that downregulation of LGR4 inhibited SCAPs proliferation, delayed migration and arrested cell cycle progression at the S and G2/M phases. ALP staining revealed that blockade of LGR4 decreased ALP activity. QRT-PCR and Western blot analysis demonstrated that LGR4 silencing reduced the expression of odonto/osteogenic markers (RUNX2, OSX, OPN, OCN and DSPP). Further Western blot and immunofluorescence studies clarified that inhibition of LGR4 disrupted β-catenin stabilization. Taken together, downregulation of LGR4 gene expression inhibited SCAPs proliferation, migration and odonto/osteogenic differentiation by blocking the Wnt/β-catenin signaling pathway. These results indicate that LGR4 might play a vital role in SCAPs proliferation and odontoblastic differentiation.

The way teeth grow is recorded in dental enamel as incremental marks. Detailed analysis of tooth growth is known to provide valuable insights into the growth and the pace of life of vertebrates. Here, we study the growth pattern of the first lower molar in several extant and extinct species of Equus and explore its relationship with life history events. Our histological analysis shows that enamel extends beyond the molar's cervix in these mammals. We identified three different crown developmental stages (CDS) in the first lower molars of equids characterised by different growth rates and likely to be related to structural and ontogenetic modifications of the tooth. Enamel extension rate, which ranges from ≈400 μm/d at the beginning of crown development to rates of ≈30 μm/d near the root, and daily secretion rate (≈17 μm/d) have been shown to be very conservative within the genus. From our results, we also inferred data of molar wear rate for these equids that suggest higher wear rates at early ontogenetic stages (13 mm/y) than commonly assumed. The results obtained here provide a basis for future studies of equid dentition in different scientific areas, involving isotope, demographic and dietary studies.

Objectives: To determine the pulp vitality after traumatic injury, dentists often use pulp sensitivity tests, which can be ambiguous in young permanent teeth with incomplete root formation. Transmitted-light plethysmography (TLP) is a non-invasive objective method that uses a 525-nm LED to detect blood volume change in the pulp. The present study aimed (1) to investigate pulpal blood flow with TLP and optical characteristics in healthy permanent maxillary incisors in different root formation stages, and (2) to assess the influences of body growth of the children and tooth color on the TLP amplitude., Design: Seventy-eight fully erupted maxillary central incisors were divided into four groups, according to the root formation stages. Group 1: root with wide-open apex, Group 2: root completed in length with open apex, Group 3: root with half-closed apex, Group 4: root with complete formation. The TLP amplitude, optical density, electric pulp testing, and cervical tooth color measurements of each group were compared using a one-way analysis of variance followed by the Bonferroni method. The correlation between the weights/heights of children and TLP amplitudes was analyzed using Pearson coefficient., Results: The TLP amplitude was significantly higher in Group 3 than in the other groups. The amplitude was correlated with the weights/heights of children, but not with the tooth color. Optical density and electric sensitivity increased with tooth maturation., Conclusion: The amplitude of TLP and optical density may be affected by growth and development in children and indicate changes in the vascular dynamics of the pulp and hard tissue maturation during root formation stages., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Aim: To investigate the response of immature permanent teeth with nonvital pulps after revitalization procedures using cone beam computed tomography imaging and conventional radiograph., Methodology: Clinical records of patients who had undergone revitalization between 1997 and 2014 were collected. Seventeen patients attended the follow-up examination. Clinical examination and radiographic examination by means of CBCT imaging and conventional radiographs were conducted to evaluate treatment outcomes, especially the dimensional changes of the root and the pattern of root maturation. Statistical analysis was conducted using Fisher's exact test and Monte Carlo test to investigate the effect of potential outcome predictors., Results: For the follow-up period as far as 96 months, thirteen teeth (76%) were classified as a 'success'. Conventional radiographs showed various degrees of dimensional changes in the immature roots ranging from 0% to 73% changes. The mean percentage changes of root length and root wall thickness were 14.38 ± 20.92% and 13.82 ± 11.17%, respectively. CBCT images illustrated various patterns of root maturation including incomplete root development with wide-open apices and fully formed roots with various apical shapes. Root canal contents varied radiographically from empty canals to many degrees of unpredictable patterning of radiopaque deposits., Conclusion: Revitalization in immature teeth with nonvital pulps resulted in unpredictable responses in terms of continued root development. Various types of root maturation and root canal contents were observed radiographically, which were different from typical root development., (© 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Signaling pathways are used reiteratively in different developmental processes yet produce distinct cell fates through specific downstream transcription factors. In this study, we used tooth root development as a model with which to investigate how the BMP signaling pathway regulates transcriptional complexes to direct the fate determination of multipotent mesenchymal stem cells (MSCs). We first identified the MSC population supporting mouse molar root growth as Gli1 + cells. Using a Gli1-driven Cre-mediated recombination system, our results provide the first in vivo evidence that BMP signaling activity is required for the odontogenic differentiation of MSCs. Specifically, we identified the transcription factors Pax9, Klf4, Satb2 and Lhx8 as being downstream of BMP signaling and expressed in a spatially restricted pattern that is potentially involved in determining distinct cellular identities within the dental mesenchyme. Finally, we found that overactivation of one key transcription factor, Klf4, which is associated with the odontogenic region, promotes odontogenic differentiation of MSCs. Collectively, our results demonstrate the functional significance of BMP signaling in regulating MSC fate during root development and shed light on how BMP signaling can achieve functional specificity in regulating diverse organ development., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2017. Published by The Company of Biologists Ltd.)

Objective: The results in human sex chromosome aneuploidies had shown that the extra Y chromosome increases permanent and deciduous tooth crown sizes in the mesiodistal and labiolingual directions. The hypothesis of the study was that the additional Y chromosome increases the permanent tooth crown growth in a vertical dimension. We also aimed to observe possible sex difference in the permanent tooth crown height., Material and Methods: Data on 15 47,XYY males or males with an extra Y chromosome, nine male relatives (five brothers and four fathers) and 45 male and 48 female population controls had been gathered previously for Professor Lassi Alvesalo's KVANTTI Research Project. The measurements from panoramic radiographs were performed of all the applicable teeth, except the third molars on both sides of the jaws with a sliding digital calliper., Results: All the mean tooth crown heights in the 47,XYY males were larger than in the male population controls and the differences were statistically significant in six teeth in the maxilla and 10 teeth in the mandible. Apart from few teeth, the crown heights in the 47,XYY males were larger than in their male relatives, but the difference between these groups was significant only in one tooth. The differences between sexes were statistically significant in eight teeth in the maxilla., Conclusions: Based on previous investigations and this work, it is evident that the impact of the extra Y chromosome during tooth crown development is holistic, increasing permanent tooth sizes in three dimensions in a balanced manner.

Objective: F-spondin is an extracellular matrix (ECM) protein that belongs to the thrombospondin type I repeat superfamily and is a negative regulator of bone mass. We have previously shown that f-spondin is specifically expressed in the dental follicle (DF), which gives rise to the periodontal ligament (PDL) during the tooth root formation stage. To investigate the molecular mechanism of PDL formation, we investigated the function of f-spondin in DF differentiation., Design: The expression patterning of f-spondin in the developing tooth germ was compared with that of periodontal ligament-related genes, including runx2, type I collagen and periostin, by in situ hybridization analysis. To investigate the function of f-spondin during periodontal ligament formation, an f-spondin adenovirus was infected into the bell stage of the developing tooth germ, and the effect on dental differentiation was analyzed., Results: F-spondin was specifically expressed in the DF of the developing tooth germ; by contrast, type I collagen, runx2 and periostin were expressed in the DF and in the alveolar bone. F-spondin-overexpresssing tooth germ exhibited a reduction in gene expression of periostin and type I collagen in the DF. By contrast, the knockdown of f-spondin in primary DF cells increased the expression of these genes. Treatment with recombinant f-spondin protein functionally inhibited periostin expression induced by transforming growth factor-β (TGF-β)., Conclusion: Our data indicated that f-spondin inhibits the differentiation of DF cells into periodontal ligament cells by inhibiting TGF-β. These data suggested that f-spondin negatively regulates PDL differentiation which may play an important role in the immature phenotype of DF., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Hypofunctional occlusion is known to lead to changes in the length of roots over time. The mechanisms that drive such changes, however, are poorly understood, with most studies concentrating on juvenile rats prior to the arrest of root development. In this article, we investigated the response of the upper and lower first molar roots to lack of occlusion concentrating on time-points after the development of the roots has ceased using the mouse as a model. Mouse molar roots finish development at weaning, much earlier than rat molars, and display a similar pattern of roots in the lower and upper jaw to humans., Methods: Hypofunctional occlusion was achieved in adult mice at 5 and 9 weeks of age by flattening the cusps of the upper first molar. Mice were then sacrificed after 6 and 2 weeks, respectively, along with control littermates. microCT was used to measure root length, alveolar bone height, and the amount of tooth eruption, followed by sectioning to understand the mechanisms behind the changes at the histological level., Results: In the lower first molar, the response to hypofunctional occlusion was characterized by elongation of both the mesial root and its surrounding alveolar bone, while the distal root was unaffected. In contrast, the response of the upper first molar was characterized by over-eruption of the mesial side of the tooth without any significant change in the alveolar bone or root length. From histologic sections, it was clear that increased deposition of cellular cementum played an important role in the changes that occurred in the lower mesial root., Conclusions: In a mouse model, upper and lower molars responded differently to hypofunctional occlusion, with adult mice showing a different response to that previously reported for juvenile rats, highlighting the importance of considering age and tooth position in cases of hypofunction., (© The Author 2016. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com)

Dental age estimation was recognized as an imperative issue in clinical and medico-legal practice. However, very few studies on dental age estimation in children have been published in Colombia. This study evaluated the accuracy of Cameriere's method of measurement of open apices on tooth roots in a sample of 526 digital panoramic radiographs (OPTs) of children (274 boys and 252 girls), aged between 6 and 14 years, from Bogotá, Valle del Cauca, Buga and Villavicencio, in Southwest Colombia. Only first seven permanent lower teeth, except third molar, from the left side of mandible were studied. Difference between dental age and chronological age was evaluated for boys and girls across nine age classes. Intra-class correlation coefficient and Kappa score was used to test intra- and inter-observer agreement error rate. Dental age was overestimated by 0.08 years and standard deviation (±SD) of 0.68 years in boys which was not statistically significant (p = 0.06), while in girls dental age underestimated by -0.25 ± 0.65 years which was statistically significant difference (p < 0.001). The absolute difference between DA and CA or mean prediction error (ME) was 0.57 ± 0.38 years in boys and 0.57 ± 0.41 in girls (p = 0.966) which implies that Cameriere's European formula is similarly accurate in both sexes in this sample of Colombian children., (Copyright © 2017 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.)

Background: Structural anomalies of teeth are observed at high rates in childhood cancer survivors (CCS). Several therapeutic exposures have been shown to be associated with dental developmental disturbances. This study was conducted to analyze the risk factors for dental developmental abnormality (DDA) and investigate the association between age at the time of cancer treatment and DDA in CCS., Patients and Methods: Fifty-six CCS were enrolled. Orthopantomography and dental examination were performed in all the patients. We evaluated the prevalence of DDA and analyzed the risk factors for each type of DDA., Results: DDAs were observed in 46.4% of CCS, including hypodontia in 9 (16.1%), abnormal roots in nine (16.1%), enamel defects/hypoplasia in 6 (10.7%), and microdontia in 12 (21.4%) patients. The number of patients with abnormal roots was significantly higher in the group treated with stem cell transplantation or at an age older than 4 years. We observed that the formation period of abnormal teeth coincided with the treatment period in the majority of CCS with DDA., Conclusions: Particularly regarding the root abnormality, treatment at elder age may be a risk factor for root developmental disturbances. Risk evaluation, appropriate follow-up, and early detection of dental issues are required for all CCS.

Dental Mesenchymal Cells (DMCs) are known to play a role in tooth development as well as in the repair and regeneration of dental tissue. A large number of signaling molecules regulate the proliferation and differentiation of DMC, though the underlying mechanisms are still not fully understood. Sirtuin-6 (SIRT6), a key regulator of aging, can exert an impact on embryonic stem cell (ESC) differentiation. The experimental deletion of Sirt6 in mouse bone marrow cells has been found to have an inhibiting impact on the bone mineral density and the osteogenic differentiation of these cells. The possible role of Sirt6 in tooth development, however, has at present remained largely unexplored. In the present study, we found that SIRT6 had no effect on tooth development before birth. However, Sirt6 gene deletion in knockout mice did have two post-natal impacts: a delay in tooth eruption and sluggishness in the development of dental roots. We propose an explanation of the possible molecular basis of the changes observed in Sirt6-/- mice. SIRT6 is expressed in mouse odontoblasts. Sirt6 deletion enhanced the proliferation of DMCs, as well as their capacity for adipogenic differentiation. On the other hand, it inhibited their capacity for in vitro osteogenic/chondrogenic differentiation. Further studies suggested that other factors may mediate the role of Sirt6 in odontogenesis. These include the nuclear factor kappa B (NF-κB), p38 mitogen-activated protein kinase (p38-MAPK), extracellular regulated MAP kinase (ERK) pathways and the mitochondrial energy. We demonstrated that Sirt6 plays a role in tooth root formation and confirmed that SIRT6 is necessary for DMC differentiation as well as for the development of the tooth root and for eventual tooth eruption. These results establish a new link between SIRT6 and tooth development.

The final stage of third molar development is used to assign a subject to above the 18-year threshold. Some subjects exhibiting this final stage are less than 18 years. Radiographs from 1000 females and 1000 males age 16.00-25.99 years were examined. Each half-year age band comprised 50 females and 50 males. Three categories of root canal widths (RCW) of the LL6, LL7, and LL8 [FDI 36, 37, and 38] were defined. Reproducibility was achieved by re-assessing the same subjects 12 months apart. For females, the minimum value for RCW-A was 16.33 years, RCW-B 17.23 years, and RCW-C 18.45 years. For males, the minimum values were RCW-A 17.16 years, RCW-B 18.29 years, and RCW-C 18.16 years. The presence of RCW-C in a female, and the presence of RCW-B or RCW-C in a male is compelling evidence that the subject is above the 18-year threshold., (© 2016 American Academy of Forensic Sciences.)

Introduction: This case report presents the treatment of a 16-year-old boy with a maxillary lateral incisor (tooth #10) presenting with Oehlers type II dens invaginatus and diagnosed with previously initiated therapy and asymptomatic apical periodontitis., Methods: A regenerative endodontic procedure (REP) was performed for the tooth but complicated by apically displaced mineral trioxide aggregate (MTA). Clinical and radiographic examination was undertaken yearly, and a cone-beam computed tomography scan was taken to investigate further the formation of hard tissues within the root canal. Subsequently, tooth #10 was re-accessed and then root-filled with MTA., Results: There was complete periapical healing, thickening of the dentinal root walls, and completed apex formation 3 years after REP. Hard tissue formation was noted within the root canal, on the root canal wall, and the root apex through clinical and radiographic examination. Less hard tissue formation was noted on the labial root canal wall where the displaced MTA was located, which was identified on the cone-beam computed tomography scan., Conclusions: This report demonstrates that REP can potentially provide excellent treatment outcomes for structurally compromised teeth. REP should be considered as a first-line treatment before proceeding with a root filling when root development is incomplete, but attention to technical detail is essential., (Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

The tooth root is an integral, functionally important part of our dentition. The formation of a functional root depends on epithelial-mesenchymal interactions and integration of the root with the jaw bone, blood supply and nerve innervations. The root development process therefore offers an attractive model for investigating organogenesis. Understanding how roots develop and how they can be bioengineered is also of great interest in the field of regenerative medicine. Here, we discuss recent advances in understanding the cellular and molecular mechanisms underlying tooth root formation. We review the function of cellular structure and components such as Hertwig's epithelial root sheath, cranial neural crest cells and stem cells residing in developing and adult teeth. We also highlight how complex signaling networks together with multiple transcription factors mediate tissue-tissue interactions that guide root development. Finally, we discuss the possible role of stem cells in establishing the crown-to-root transition, and provide an overview of root malformations and diseases in humans., (© 2017. Published by The Company of Biologists Ltd.)

Age estimation is an important procedure in forensic medicine and is carried out for a number of reasons. For living persons, age estimation is performed in order to assess whether a child has attained the age of criminal responsibility, in scenarios involving rape, kidnapping or marriage, in premature births, adoption procedures, illegal immigration, pediatric endocrine diseases and orthodontic malocclusion, as well as in circumstances in which the birth certificate is not available or the records are suspect. According to data from the UNHCR (United Nations High Commissioner for Refugees), the number of people seeking refugee status continued to increase in the last years, driven by the wars in Syria and Iraq, as well as by conflict and instability in Afghanistan, Eritrea and elsewhere. The objective of this study is to compare the accuracy of estimating dental age versus chronological age using the Nolla and Demirjian methods in a Spanish population. A final sample of 2641 panoramic X-rays corresponding to Spanish patients (1322 males and 1319 females) between 7-21 years of age was analyzed. Dental age was assessed using the Nolla and Demirjian methods, establishing comparisons with mean chronological age based on the Student t-test for paired samples, followed by the generation of a linear regression model. Both methods showed slight discrepancy between dental and chronological age. On examining the reproducibility of the Nolla and Demirjian methods, technical errors of 0.84% and 0.62%, respectively, were observed. On average, the Nolla method was found to estimate an age 0.213years younger than the chronological age, while the Demirjian method estimated an age 0.853years older than the chronological age. Linear combination of the mean Nolla and Demirjian estimates increased the predictive capacity to 99.2%. In conclusion the Nolla and Demirjian methods were found to be accurate in estimating chronological age from dental age in a Spanish population. The error was found to be greater in males than in females, and involved an over-estimation of age with the Demirjian method and under-estimation of age with the Nolla method. Combination of the Nolla and Demirjian methods for estimating chronological age from dental age affords a predictive capacity of over 99%, and is fast and easy to perform, and inexpensive., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Introduction: Mutations in the proteinase bone morphogenetic protein-1 (BMP1) were recently identified in patients with osteogenesis imperfecta, which can be associated with type 1 dentinogenesis imperfecta. BMP1 is co-expressed in various tissues and has overlapping activities with the closely related proteinase mammalian tolloid-like 1 (TLL1). In this study we investigated whether removing the overlapping activities of BMP1 and TLL1 affects the mineralization of tooth root dentin., Methods: Floxed alleles of the BMP1 and TLL1 genes were excised via ubiquitously expressed Cre induced by tamoxifen treatment beginning at 3 days of age (harvested at 3 weeks of age) or beginning at 4 weeks of age (harvested at 8 weeks of age). Multiple techniques, including x-ray analysis, double-labeling with calcein and alizarin red stains for measurement of dentin formation rate, and histologic and immunostaining assays, were used to analyze the dentin phenotype., Results: BMP1/TLL1 double knockout mice displayed short and thin root dentin, defects in dentin mineralization, and delayed tooth eruption. Molecular mechanism studies revealed accumulation of collagens in dentin and a sharp reduction in non-collagenous proteins such as dentin matrix protein 1 and dentin sialophosphoprotein. Furthermore, we found a strong reduction in tartrate-resistant acid phosphatase, which is likely caused by defects in bone cells., Conclusions: BMP1/TLL1 appear to play crucial roles in maintaining extracellular matrix homeostasis essential to root formation and dentin mineralization., Competing Interests: The authors deny any conflicts of interest related to this study. This research is original and free of conflict of interest., (Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

The purpose of this paper was to explore the potential application of Root Pulp Visibility (RPV) to determine whether or not a subject of unknown date of birth is under or over the 18year threshold., Patients, Materials and Methods: Two thousand Dental Panoramic Radiographs comprising equal numbers of females and males evenly distributed between the ages of 16 and 26 years were examined. All third molars were scored using the Anglo-Canadian 8 Stage Tooth Development system [1]. The Lower Left Third Molar was censored to avoid redundant data being included in the data array for the Age at Attainment of Stage H, the final stage of tooth development (Roberts et al., 2015 [2]). A filter cascade was applied to the data to select the data array for 1. Teeth in the final stage of development, 2. Female or male gender, 3. The definitive stage of RPV: RPV-A, RPV-B, RPV-C, or RPV-D., Results: The summary data for each of these stages was derived and the minimum value for each was used to determine the relationship of the minimum value of A, B, C or D to the 18 Year threshold. It was found that for both females and males the minimum value for RPV-A and RPV-B indicated that subjects displaying this could be under 18 years old. For RPV-C and RPV-D, the minimum values show unequivocally that for both females and males the subject was over 18 years old., Conclusion: RPV has an important role in unambiguously determining whether a subject is below or above the 18year threshold., (Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.)

Congenital diseases of tooth roots, in terms of developmental abnormalities of short and thin root phenotypes, can lead to loss of teeth. A more complete understanding of the genetic molecular pathways and biological processes controlling tooth root formation is required. Recent studies have revealed that Osterix (Osx), a key mesenchymal transcriptional factor participating in both the processes of osteogenesis and odontogenesis, plays a vital role underlying the mechanisms of developmental differences between root and crown. During tooth development, Osx expression has been identified from late embryonic to postnatal stages when the tooth root develops, particularly in odontoblasts and cementoblasts to promote their differentiation and mineralization. Furthermore, the site-specific function of Osx in tooth root formation has been confirmed, because odontoblastic Osx-conditional knockout mice demonstrate primarily short and thin root phenotypes with no apparent abnormalities in the crown (Journal of Bone and Mineral Research 30, 2014 and 742, Journal of Dental Research 94, 2015 and 430). These findings suggest that Osx functions to promote odontoblast and cementoblast differentiation and root elongation only in root, but not in crown formation. Mechanistic research shows regulatory networks of Osx expression, which can be controlled through manipulating the epithelial BMP signalling, mesenchymal Runx2 expression and cellular phosphorylation levels, indicating feasible routes of promoting Osx expression postnatally (Journal of Cellular Biochemistry 114, 2013 and 975). In this regard, a promising approach might be available to regenerate the congenitally diseased root and that regenerative therapy would be the best choice for patients with developmental tooth diseases., Competing Interests: The authors have stated explicitly that there are no conflicts of interest in connection with this article., (© 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

The muscle segment homeogenes Msx1 and Msx2 play a major role in tooth and bone formation. Periodontal osteoclast impairment also occurs in Msx2 null mutant mice, which is restored by overexpression of the receptor activator of NF-κB targeted in osteoclast lineage. Here, we investigated the role of Msx2 in dentinogenesis. Experiments were performed on Msx2(-/-) mice and the MDPC-23 odontoblastic cell line. After Msx2 gene silencing, real-time quantitative RT-PCR data showed significant overexpression of Runx2, Bglap, Dspp, and Alpl. Of three inhibitors of Wnt/β-catenin signaling (Dkk1, SostDc1, and Sost/Sclerostin), only Sost was expressed in postnatal teeth and overexpressed in Msx2(-/-) tooth samples. Initial crown dentin formation-primary dentinogenesis-occurred fairly normally in Msx2(-/-) teeth, albeit with distorted cusp patterns. Later stages of tooth development were characterized by a deviation from secondary toward tertiary dentinogenesis with osteodentin formation and impaired dentin deposition leading to limited root elongation. In Msx2(-/-)/receptor activator of NF-κB-transgenic double mutants, the dentin phenotype, notably in the roots, was rescued and sclerostin levels were normalized. These data suggest that Msx2 may act indirectly on dentinogenesis by controlling osteoclast activity and the signaling network related to eruption, supporting and further extending the concept that Msx2 controls formation of mineralized tissues by inhibition of the Wnt/β-catenin pathway; Sost in dentin and Dkk1 in bone, as previously demonstrated., (Copyright © 2016 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)

Objective: Ghrelin, an appetite-stimulating hormone, plays diverse regulatory functions in cell growth, proliferation, differentiation and apoptosis during mammalian development. There is limited information currently available regarding Ghrelin expression during mammalian tooth development, thus we aimed to establish the spatiotemporal expression of Ghrelin during murine molar odontogenesis., Design: Immunohistochemistry was performed to detect the expression pattern of Ghrelin in mandible molar from E15.5 to PN7 during murine tooth development., Results: The results showed that Ghrelin initially expressed in the inner enamel epithelium and the adjacent mesenchymal cells below, further with persistent expression in the ameloblasts and odontoblasts throughout the following developmental stages. In addition, Ghrelin was also present in Hertwig's epithelial root sheath at the beginning of tooth root formation., Conclusions: These results suggest that Ghrelin was present in tooth organs throughout the stages of tooth development, especially in ameloblasts and odontoblasts with little spatiotemporal expression differences. However, the potential regulatory roles of this hormone in tooth development still need to be validated by functional studies., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This article deals with the unusual course of failed revascularization/revitalization cases. Up to this date the evidence of success and failure rates of revascularization/revitalization treatment is scarce. These case reports present "unsuccessful" revascularization/revitalization treatment of permanent immature teeth with apical periodontitis. Although the teeth were treated by protocol suggested by the American Association of Endodontists and the symptoms disappeared, maturation of teeth continued, and periapical lesion was reduced, the teeth went symptomatic during the follow-up. Subsequently, regular root canal treatment was performed. Despite chronic infection that was probably left inside the root canal after a disinfection protocol, the secondary aims of the treatment were achieved even when the primary ones were not. The possible causes of failures of revascularization/revitalization treatment and their prevention are discussed., (Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Membrane-type matrix metalloproteinase 1 (MT1-MMP) is a transmembrane zinc-endopeptidase that breaks down extracellular matrix components, including several collagens, during tissue development and physiological remodeling. MT1-MMP-deficient mice (MT1-MMP(-/-)) feature severe defects in connective tissues, such as impaired growth, osteopenia, fibrosis, and conspicuous loss of molar tooth eruption and root formation. In order to define the functions of MT1-MMP during root formation and tooth eruption, we analyzed the development of teeth and surrounding tissues in the absence of MT1-MMP. In situ hybridization showed that MT1-MMP was widely expressed in cells associated with teeth and surrounding connective tissues during development. Multiple defects in dentoalveolar tissues were associated with loss of MT1-MMP. Root formation was inhibited by defective structure and function of Hertwig's epithelial root sheath (HERS). However, no defect was found in creation of the eruption pathway, suggesting that tooth eruption was hampered by lack of alveolar bone modeling/remodeling coincident with reduced periodontal ligament (PDL) formation and integration with the alveolar bone. Additionally, we identified a significant defect in dentin formation and mineralization associated with the loss of MT1-MMP. To segregate these multiple defects and trace their cellular origin, conditional ablation of MT1-MMP was performed in epithelia and mesenchyme. Mice featuring selective loss of MT1-MMP activity in the epithelium were indistinguishable from wild type mice, and importantly, featured a normal HERS structure and molar eruption. In contrast, selective knock-out of MT1-MMP in Osterix-expressing mesenchymal cells, including osteoblasts and odontoblasts, recapitulated major defects from the global knock-out including altered HERS structure, short roots, defective dentin formation and mineralization, and reduced alveolar bone formation, although molars were able to erupt. These data indicate that MT1-MMP activity in the dental mesenchyme, and not in epithelial-derived HERS, is essential for proper tooth root formation and eruption. In summary, our studies point to an indispensable role for MT1-MMP-mediated matrix remodeling in tooth eruption through effects on bone formation, soft tissue remodeling and organization of the follicle/PDL region., (Copyright © 2016 International Society of Matrix Biology. All rights reserved.)

Dental root formation is a dynamic process in which mesenchymal cells migrate toward the site of the future root, differentiate and secrete dentin and cementum. However, the identities of dental mesenchymal progenitors are largely unknown. Here we show that cells expressing osterix are mesenchymal progenitors contributing to all relevant cell types during morphogenesis. The majority of cells expressing parathyroid hormone-related peptide (PTHrP) are in the dental follicle and on the root surface, and deletion of its receptor (PPR) in these progenitors leads to failure of eruption and significantly truncated roots lacking periodontal ligaments. The PPR-deficient progenitors exhibit accelerated cementoblast differentiation with upregulation of nuclear factor I/C (Nfic). Deletion of histone deacetylase-4 (HDAC4) partially recapitulates the PPR deletion root phenotype. These findings indicate that PPR signalling in dental mesenchymal progenitors is essential for tooth root formation, underscoring importance of the PTHrP-PPR system during root morphogenesis and tooth eruption.

Introduction: Our aim in this study was to investigate the association between dental mineralization stages and the periods of the pubertal growth spurt (PGS)., Methods: The sample included panoramic and hand-wrist radiographs from 491 subjects (222 boys, 269 girls) aged 7 to 17 years. Dental development was rated, and skeletal maturation was evaluated. The relevant associations were investigated by analysis of ordinal multinomial logistic regression., Results: The second molar (odds ratio [OR] = 4.34) and the first premolar (OR = 2.45) were the best growth predictors for girls. For boys, the second molar (OR = 6.80), second premolar (OR = 2.41), and canine (OR = 3.21) proved to be the best predictors. Stages D and E of the second molar for girls, and stages E and F for boys, corresponded to the onset of the accelerated growth spurt. Stage F of the second molar for girls and stage G for boys corresponded to the peak of the PGS. At the end of the PGS, most teeth had already attained apical closure. In girls, however, most second molars were found at stage G., Conclusions: An association exists between the dental mineralization stages and the periods of the PGS, especially for second molars. Panoramic radiographs can be used as the first diagnostic tool to estimate the pubertal growth period., (Copyright © 2016 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: The purpose of this study was to evaluate the effectiveness of autologous platelet concentrates (APCs) in promoting pulp and dentin regeneration in animal models., Methods: An electronic search was performed on MEDLINE, Embase, Scopus, SciELO, LILACS, and CENTRAL. Animal studies using APC as a root filling material after pulpectomy in mature or immature teeth were included. Articles underwent risk of bias assessment. Histologic evaluation of intracanal neoformed tissue was the primary outcome; root development, root wall thickening, apical closure, and periapical healing in apical periodontitis were the secondary outcomes., Results: Seven articles were included. Platelet-rich plasma (PRP) was used as root filling material during regenerative procedures in the experimental group in either mature or immature teeth. After revascularization with PRP alone or in conjunction with stem cells of a different source, the histologic analyses revealed that, in addition to an odontoblastic cell layer or dentinlike structure, the neoformed intracanal tissues were mainly cementumlike, bonelike, and connective tissues., Conclusions: True regeneration of necrotic pulp may not be achieved with current techniques using PRP, all of which stimulated tissue repair. Benefits of PRP adjunct for pulp tissue regeneration in preclinical studies remain unclear. Further studies with standardized protocols are necessary to assess the actual contribution of PRP in endodontic regenerative therapies., (Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Regenerative endodontic treatment has provided a treatment option that aims to allow root maturation. The present report describes the regenerative endodontic treatment of a necrotic, immature molar by using recombinant human platelet-derived growth factor (rhPDGF-BB) and shows the continued root maturation in the tooth with arrested root development. A regenerative endodontic procedure that used a growth factor was performed for a necrotic molar with arrested root formation in a 20-year-old patient. Thorough disinfection by using mechanical instrumentation and copious irrigation of antimicrobial agents as well as intracanal medication with calcium hydroxide was performed throughout the first 2 appointments. At the third appointment, the root canals were irrigated with an antimicrobial solution and 17% EDTA, and bleeding was evoked by passing sterile paper points beyond the apex in each canal. Small pieces of a collagen membrane saturated with rhPDGF-BB solution from GEM 21S were packed into each canal. Mineral trioxide aggregate was placed, and Cavit and composite resin were used to restore the tooth. Complete root maturation and resolution of a periapical radiolucency were observed at the 15-month follow-up. The present report presents a regenerative endodontic procedure that uses rhPDGF-BB for a necrotic molar with arrested root development. The finding of continued root development in the present case suggests that regenerative endodontic treatment may be able to resume the root maturation process in teeth with arrested root formation. Further clinical studies are required to investigate the efficacy of rhPDGF-BB in regenerative endodontic treatment., (Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

c-Fos homozygous mice lack osteoclasts with a failure of the teeth to erupt and with an arrest of root development. Here, we characterize the defects associated with the failure in root development and the loss of the tooth-bone interface, and we investigate the underlying causes. We show that, while homozygous c-Fos mice have no multinucleated osteoclasts, heterozygous mice have a reduction in the number of osteoclasts with a reduction in the tooth-bone interface during development and subtle skeletal defects postnatally. In the homozygous mutants bone is found to penetrate the tooth, particularly at the apical end, physically disrupting the root forming HERS (Hertwig's epithelial root sheath) cells. The cells of the HERS continue to proliferate but cannot extend downward due to the presence of bone, leading to a loss of root formation. Tooth germ culture showed that the developing tooth invaded the static bone in mutant tissue, rather than the bone encroaching on the tooth. Although c-Fos has been shown to be expressed in developing teeth, the defect in maintenance of the tooth-bone interface appears to be driven solely by the lack of osteoclasts, as this defect can be rescued in the presence of donor osteoclasts. The rescue suggests that signals from the tooth recruit osteoclasts to clear the bone from around the tooth, allowing the tooth to grow, form roots, and later erupt., (© International & American Associations for Dental Research 2015.)

Introduction: Autotransplantation has become a major therapeutic option for replacing missing teeth in adult orthodontic patients. However, little systematic information is available about the long-term stability of autotransplanted teeth with complete root formation after the application of an orthodontic force. The objective of this study was to investigate the outcomes of autotransplanted teeth with complete root formation that underwent orthodontic treatment., Methods: One hundred teeth, autotransplanted in 89 patients, were examined over a mean observation period of 5.8 years. Orthodontic force was applied with nickel-titanium wires 4 to 8 weeks after autotransplantation. Root resorption, ankylosis, mobility, pocket depth, and inflammation at the recipient site were investigated clinically and with radiographs., Results: The survival rate of the autotransplanted teeth was 93.0%. Abnormal findings were found in 29 teeth, including 7 lost teeth, for a success rate of 71.0%. Donor tooth type and occlusal condition of the donor tooth before transplantation were associated with abnormal findings., Conclusions: The early application of orthodontic force may increase the success rate of autotransplanted teeth, and the type and presurgical occlusal condition of donor teeth affect the success rate., (Copyright © 2015 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Background: Apexogenesis after traumatic exposure in vital young permanent teeth can be accomplished by implementing the appropriate vital pulp therapy such as pulp capping (direct or indirect) or pulpotomy (partial or complete) depending on the time between the trauma and treatment of the patient, degree of root development, and size of the pulp exposure., Case Report: Two children with respectively 2 and 1 complicated enamel dentine fractures in immature permanent incisors were treated with new tricalcium silicate cement (Biodentine™). The treatment plan in these cases was to maintain pulp vitality aiming for apexogenesis which allows continued root development along the entire root length. Endodontic management included partial pulpotomy or pulpotomy using Biodentine™. Clinical and radiographical evaluation (up to 48 months) showed continual apexogenesis with no periodontal or periapical pathology. The appropriate restorations were functionally acceptable and aesthetically satisfying. The three traumatised teeth showed complete success both clinically (vitality and aesthetic outcome) as well as radiographically (apexogenesis and absence of pathological findings) after up to 48 months follow-up., Conclusion: Biodentine™ is a suitable alternative to MTA for vital pulpotomy in traumatised permanent incisors. It is also beneficial as a temporary filling without any risk of discolouration.

Hutchinson-Guilford progeria syndrome is an extremely rare condition classified as one of the premature ageing syndromes. This case presents a 16-year-old Israeli female patient, suffering from a variant of Hutchinson-Guilford progeria with a history of treatment with oral biphosphnates. The patient presented with typical cranial and facial features of the syndrome including delayed teeth eruption and root development probably due to insufficient jaw growth and severs retrognatic position of the maxilla and mandible. Orthodontic treatment considerations are described along with those required in light of the previous treatment by oral biphosphonates.All primary teeth were extracted in three appointments while creating as minimal trauma as possible to the surrounding tissue and alveolar bone. For now, the patient refuses to begin the orthodontic treatment course. There are no limitations to conduct any dental procedures in progeria patients, however, extreme caution must be exercised during oral surgery due to the inelasticity of tissues and dermal atrophy. Orthodontic procedure commencement should be early enough to manage the delayed development and eruption of teeth. Patients taking oral biphosphonates should be advised of this potential complication. If orthodontic treatment is considered appropriate, plans should be assessed and modified to include compromises.

Autotransplants are often applied in the Scandinavian countries. The indication for treatment concerns especially young patients for whom teeth with open apices are moved to areas with ageneses. The great advantage is the vital periodontal ligament, by means of which these teeth are able to grow along. In Rotterdam, during the past 11 years, transplants of teeth with fully developed apices have been carried out in cases with comparable indications. Over short and long periods of time, the results have been the same as those in the published findings in the literature. The fully developed apices appear to be just as successful as the open apices. The difference with the results in the research literature concerns the indications for transplantation in particular. These are not limited to the molar region. Within the same indication, the results are better by comparison with the implants, in both the transplants with open and with closed apices. On the basis of these results, autotransplants in dentistry have become an indication for every age group and when possible, a better treatment option than implants.

Coordination between the Hertwig's epithelial root sheath (HERS) and apical papilla (AP) is crucial for proper tooth root development. The hedgehog (Hh) signaling pathway and Nfic are both involved in tooth root development; however, their relationship has yet to be elucidated. Here, we establish a timecourse of mouse molar root development by histological staining of sections, and we demonstrate that Hh signaling is active before and during root development in the AP and HERS using Gli1 reporter mice. The proper pattern of Hh signaling activity in the AP is crucial for the proliferation of dental mesenchymal cells, because either inhibition with Hh inhibitors or constitutive activation of Hh signaling activity in transgenic mice leads to decreased proliferation in the AP and shorter roots. Moreover, Hh activity is elevated in Nfic(-/-) mice, a root defect model, whereas RNA sequencing and in situ hybridization show that the Hh attenuator Hhip is downregulated. ChIP and RNAscope analyses suggest that Nfic binds to the promoter region of Hhip. Treatment of Nfic(-/-) mice with Hh inhibitor partially restores cell proliferation, AP growth and root development. Taken together, our results demonstrate that an Nfic-Hhip-Hh signaling pathway is crucial for apical papilla growth and proper root formation. This discovery provides insight into the molecular mechanisms regulating tooth root development., (© 2015. Published by The Company of Biologists Ltd.)

Objective: The aim of this study was to evaluate 1. the differences among the cleft sides of unilateral cleft lip and palate (UCLP) patients, non-cleft sides of the same UCLP patients and well matched control patients in the root development and position of permanent upper central incisors and 2. possible gender differences., Materials and Methods: The study sample consisted of 40 patient (20 males and 20 females; mean age: 14.84±3.2 years) with non-syndromic UCLP patients, and 40 control patients (20 males and 20 females; mean age: 13.38±1.6 years). Cone-beam computed tomography (CBCT) images were taken off from all patients. All tomographs were obtained in supine position by using CBCT (NewTom 5G, QR, Verona, Italy)., Results: Significant smaller root volume of central incisor was observed on the cleft side than on the non-cleft side of UCLP patients (P < 0.05). No significant difference in the root development and position of the central incisors was found between the non-cleft side of the UCLP and the control group (P > 0.05). There were no statistically significant gender differences in any linear, angular, or volumetric measurements (P > 0.05). Only the root volume of the cleft side was smaller in females than in males (P < 0.05)., Conclusions: In general, root volumes of central incisors on the cleft side were 12.15 per cent smaller than non-cleft side. Root development of the central incisor was much more influenced by the cleft in females than in males., (© The Author 2014. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Introduction: In this study, we evaluated root and alveolar bone development in unilateral osseous impacted immature maxillary central incisors by cone-beam computed tomography before and after closed-eruption treatment, in comparison with naturally erupted contralateral immature maxillary central incisors., Methods and Results: The study included 30 patients, 20 boys and 10 girls, with a mean age of 8.44 ± 1.20 years (range, 6.5-11.2 years). After treatment, the root lengths of both the impacted maxillary central incisors (10.66 ± 2.10 mm) and the contralateral maxillary central incisors (11.04 ± 1.76 mm) were significantly greater than their pretreatment values (6.67 ± 1.94 and 9.02 ± 2.13 mm, respectively). The root canal widths of the incisors decreased significantly after treatment. From the posttreatment cone-beam computed tomography images, the ratio of exposed root length to total root length and the thickness of the alveolar bone at 1 mm under the alveolar crest and at the apex were calculated to evaluate alveolar bone development. Impacted immature maxillary central incisors differed significantly from contralateral immature maxillary central incisors in labial exposed root length, labial ratio to total root length, and lingual alveolar crest. Clinical crown height was higher (statistically but not clinically) for the impacted incisors (9.87 mm) than for the contralateral incisors (9.37 mm)., Conclusions: Impacted immature incisors grew to the same stage as did erupted contralateral incisors after closed-eruption treatment. Both incisor types had some alveolar bone loss, and thin alveolar bone surrounded the roots., (Copyright © 2015 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Background: Survivors of childhood cancer are at risk of late dental development. Cyclophosphamide is one of the most commonly used chemotherapeutic agents against cancer in children. The aim of this study was to investigate the effects of cyclophosphamide on root formation in the molars of growing mice and to assess the morphological changes in these roots using three-dimensional structural images., Methods: We treated 16 12-day-old ICR mice with cyclophosphamide (100 mg/kg, i.p.) and 16 control mice with saline. At 16, 20, 24, and 27 days of age, the mandibular left first molars were scanned using soft micro-computed tomography. After scanning, the structural indices were calculated using a three-dimensional image analysis system, and the images were subjected to three-dimensional reconstruction. The length and apical foramen area of all distal roots were assessed. Histological changes in the apical region were then assessed via hematoxylin and eosin staining., Results: The mandibular molars of all experimental mice showed evidence of cytotoxic injury, which appeared in the form of anomalous root shapes. Although all roots developed further after cyclophosphamide injection, the three-dimensional structural images showed that the roots in the experimental group tended to develop more slowly and were shorter than those in the control group. At 27 days of age, the mean root length was shorter in the experimental group than in the control group. Conversely, the apical foramen of the roots in the experimental group tended to close faster than that of roots in the control group. In addition, hematoxylin and eosin staining of the distal roots in the experimental group showed increased dentin thickness in the apical region., Conclusion: Our results suggest that cyclophosphamide can result in short root lengths and early apical foramen closure, eventually leading to V-shaped or thin roots.

Despite increasing knowledge of the basic molecular aspects of bone formation and its regulation, the mechanisms of bone morphogenesis leading to a topologically specific shape remain unknown. The formation of the alveolar bone, which houses the dental primordia and later, the dental roots, may serve as a model to understand the formation of bone form in general. Thirty-eight heads of mice (C57 Bl/6J) ranging from stages E13-P20 were used to prepare histological serial sections. For each stage, virtual 3D-reconstructions were made in order to study the morphogenesis of the mandibular molar primordia concomitantly with their surrounding bone. Special focus was given to recording the remodeling pattern. It has been shown that, in early stages (E13, E14), bone formation is characterized by apposition only. In stage E15, the bony crypt around the dental primordia is remodeled mostly by resorption of bone. In stage E18, the bone remodeling pattern shows resorption all along the bony gutter, which houses the molar primordia. The medial and lateral margins are characterized by apposition. At birth (stage P0), a bony septum has begun to form between the primordium m1 and of m2, arising from both sides and characterized by pure apposition of bone. In stage P4, the crypts of m1 and m2, and also that of m3, show bone resorption inside, while the medial and lateral bony margins show apposition of bone throughout. Generally, during development, the bone gradually encapsulates the dental primordia, in such a way that the bone reaches over the dental primordia and leaves only a continuous longish opening of about 200μm width. The opening at the occlusal surface of m1, at the time of eruption, starting at stage P14, appears to have increased in size again. The distance between bone and dental primordium undergoes change during development. In erupted molars, it is around 100μm, during early developmental stages, it may be as less as 20μm. These data show the inevitability of bone remodeling., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Root development of permanent teeth is disturbed in survivors of childhood cancer. Cyclophosphamide (CY) is a cytostatic drug commonly used for chemotherapy in children with cancer. This study aimed to evaluate the effects of CY on the development of molar teeth until the completion of occlusion in young mice, focusing on Hertwig's epithelial root sheath (HERS). We treated thirty-two 12-day-old ICR mice with CY (100 mg/kg; 100-CY group), and 36 control mice with saline. At 12, 14, 16, 20, 24, 27, 39, 60, and 76 days of age, the mandibular molars were removed. Soft X-ray radiographs were obtained in lateral projection. The root/crown length (R/C) ratio of the first molar was calculated. Serial sagittal sections were prepared and histomorphological hematoxylin and eosin (HE) staining and immunohistochemical (cytokeratin) studies were performed. The R/C ratio of the 100-CY group (0.78) was smaller than that of the control group (1.23) at 76 days (p < 0.05, t test). While all roots developed further after injection, microscopic examination showed that the roots of the first molars that developed in the 100-CY group were shorter than those in the control group. In addition, experimental mice showed apical closure of the roots. By 20 days after injection, the HERS had disappeared from the root surface in the 100-CY group. In conclusion, this study indicates that CY can induce a defect in HERS and cause early loss of HERS. Disruption of the epithelial sheath inhibits normal root formation, and it could cause irreversible short-root development.

Traumatic injuries and dental caries can be a big challenge to immature teeth. In these cases, the main purpose of treatment is to maintain the pulp vitality. The purpose of this study was to investigate the effect of low-level laser therapy on accelerating the rate of dentinogenesis in pulpotomy of immature permanent teeth (apexogenesis). Three dogs, 4-6 months old, were used in this study. One jaw in each dog was randomly assigned to laser irradiation group. All selected teeth were pulpotomized with mineral trioxide aggregate (MTA) and restored with amalgam. In the laser group, the Ga-Al-As laser (810 nm, 0.3 W, 4 J/cm(2), 9 s) was used on buccal and lingual gingiva of each tooth in 48 h intervals for 2 weeks. In order to observe the newly formed dentine, tetracycline was injected on the 1st, 3rd, 7th, and 14th day after the operation. Then, ground sections of teeth were observed under a fluorescence microscope. The data was analyzed with Generalized Estimating Equations (GEE) test. The mean distance between the lines of tetracycline formed on the 1st and 14th day was significantly higher in the laser group (P = 0.005). Within the limitation of this study, irradiation of Ga-Al-As laser (810 nm) can accelerate the rate of dentinogenesis in apexogenesis of immature permanent teeth with MTA in dogs.

Shark and ray (elasmobranch) dentitions are well known for their multiple generations of teeth, with isolated teeth being common in the fossil record. However, how the diverse dentitions characteristic of elasmobranchs form is still poorly understood. Data on the development and maintenance of the dental patterning in this major vertebrate group will allow comparisons to other morphologically diverse taxa, including the bony fishes, in order to identify shared pattern characters for the vertebrate dentition as a whole. Data is especially lacking from the Batoidea (skates and rays), hence our objective is to compile data on embryonic and adult batoid tooth development contributing to ordering of the dentition, from cleared and stained specimens and micro-CT scans, with 3D rendered models. We selected species (adult and embryonic) spanning phylogenetically significant batoid clades, such that our observations may raise questions about relationships within the batoids, particularly with respect to current molecular-based analyses. We include developmental data from embryos of recent model organisms Leucoraja erinacea and Raja clavata to evaluate the earliest establishment of the dentition. Characters of the batoid dentition investigated include alternate addition of teeth as offset successional tooth rows (versus single separate files), presence of a symphyseal initiator region (symphyseal tooth present, or absent, but with two parasymphyseal teeth) and a restriction to tooth addition along each jaw reducing the number of tooth families, relative to addition of successor teeth within each family. Our ultimate aim is to understand the shared characters of the batoids, and whether or not these dental characters are shared more broadly within elasmobranchs, by comparing these to dentitions in shark outgroups. These developmental morphological analyses will provide a solid basis to better understand dental evolution in these important vertebrate groups as well as the general plesiomorphic vertebrate dental condition.

The topography of the auriculotemporal nerve (ATN) root system is the main criterion of this nerve classification. Previous publications indicate that ATN may have between one and five roots. Most common is a one- or two-root variant of the nerve structure. The problem of many publications is the inconsistency of nomenclature which concerns the terms "roots", "connecting branches", or "branches" that are used to identify the same structures. This study was performed on 80 specimens (40 adults and 40 fetuses) to propose a classification based on: (i) the number of roots, (ii) way of root division, and (iii) configuration of interradicular fibers that form the ATN trunk. This new classification is a remedy for inconsistency of nomenclature of ATN in the infratemporal fossa. This classification system has proven beneficial when organizing all ATN variants described in previous studies and could become a helpful tool for surgeons and dentists. Examination of ATN from the infratemporal fossa of fetuses (the youngest was at 18 weeks gestational age) showed that, at that stage, the nerve is fully developed.

Tooth is made of crown and root. It is widely believed that dentin formation in crown and root uses the same regulatory mechanism. However, identification of nuclear factor 1 C (NFIC)'s unique function in determining root but not crown dentin formation challenges the old thinking. In searching for the target molecules downstream of NFIC, we unexpectedly found a sharp reduction of osterix (OSX), the key transcription factor in skeleton formation, in the Nfic knockout (Nfic-KO) tooth root. We then demonstrated a dose-dependent increase of Osx in the odontoblast cell line due to a transient transfection of Nfic expression plasmid. Studies of global and conditional Osx-KO mice revealed no apparent changes in the crown dentin tubules and dentin matrix. However, the OSX conditional KO (cKO) mice (crossed to the 2.3-kb collagen type 1 [Col1]-Cre) displayed an increase in cell proliferation but great decreases in expressions of root dentin matrix proteins (dentin matrix protein 1 [DMP1] and dentin sialophosphoprotein [DSPP]), leading to an inhibition in odontoblast differentiation, and short, thin root dentin with few dentin tubules. Compared to the Nfic-KO tooth, which contains essentially no dentin tubules and remains in a "root-less" status at adult stages, the Osx-cKO root phenotype had partially improved at the late stage, indicating that other factors can compensate for OSX function. Thus, we conclude that OSX, one of the key downstream molecules of NFIC, plays a critical role in root, but not crown, formation., (© 2014 American Society for Bone and Mineral Research.)

The main objective of treatment of an undeveloped tooth (immature) is to provide vital pulp therapy to allow continued development of root dentin. A case report is presented that demonstrates the use of calcium hydroxide (CaOH₂) as an indirect pulp-copping material for the purpose of continued maturogenesis of an early-erupted permanent tooth with severe mobility and almost-begun root formation. Seven-year radiographic and clinical follow-up demonstrated a vital pulp and physiologic root development without any endodontic failure clinically or radiographicaly.

Wnt signaling plays an essential role in the dental epithelium and mesenchyme during tooth morphogenesis. However, it remains unclear if Wnt ligands, produced from dental mesenchyme, are necessary for odontoblast differentiation and dentin formation. Here, we show that odontoblast-specific disruption of Wntless (Wls), a chaperon protein that regulates Wnt sorting and secretion, leads to severe defects in dentin formation and root elongation. Dentin thickness decreased remarkably and pulp chambers enlarged in the mandibular molars of OC-Cre;Wls(CO/CO) mice. Although the initial odontoblast differentiation was normal in the mutant crown, odontoblasts became cuboidal and dentin thickness was reduced. In immunohistochemistry, Wnt10a, β-catenin, type I collagen, and dentin sialoprotein were significantly down-regulated in the odontoblasts of mutant crown. In addition, roots were short and root canals were widened. Cell proliferation was reduced in the developing root apex of mutant molars. Furthermore, Wnt10a and Axin2 expression was remarkably decreased in the odontoblasts of mutant roots. Deletion of the Wls gene in odontoblasts appears to reduce canonical Wnt activity, leading to inhibition of odontoblast maturation and root elongation., (© International & American Associations for Dental Research 2015.)

Bone and dentin share similar biochemical compositions and physiological properties. Dentin, a major tooth component, is formed by odontoblasts; in contrast, bone is produced by osteoblasts. Osterix (Osx), a zinc finger-containing transcription factor, has been identified as an essential regulator of osteoblast differentiation and bone formation. However, it has been difficult to establish whether Osx functions in odontoblast differentiation and dentin formation. To understand the role of Osx in dentin formation, we analyzed mice in which Osx was subjected to tissue-specific ablation under the control of either the Col1a1 or the OC promoter. Two independent Osx conditional knockout mice exhibited similar molar abnormalities. Although no phenotype was found in the crowns of these teeth, both mutant lines exhibited short molar roots due to impaired root elongation. Furthermore, the interradicular dentin in these mice showed severe hypoplastic features, which were likely caused by disruptions in odontoblast differentiation and dentin formation. These phenotypes were closely related to the temporospatial expression pattern of Osx during tooth development. These findings indicate that Osx is required for root formation by regulating odontoblast differentiation, maturation, and root elongation. Cumulatively, our data strongly indicate that Osx is a site-specific regulator in tooth root formation., (© International & American Associations for Dental Research 2015.)