Relación entre el diagnóstico histopatológico de sacos foliculares de terceros molares y la medida radiográfca estandarizada en radiografía panorámica digital / Relationship between histopathologic diagnosis of third molar follicular sacs and standardized radiographic measurement on digital panoramic radiography

Objetivo: establecer la relación entre el diagnóstico histopatológico de sacos foliculares de terceros molares y la medida radiográfca estandarizada en radiografía panorámica digital. Métodos: se llevó a cabo un estudio descriptivo en el que se incluyeron 28 sacos foliculares de terceros molares. Dos observadores midieron la radiolucidez pericoronal en radiografías panorámicas digitales usando un método estandarizado y se calculó el índice de correlación intraclase. Se estableció un diagnóstico radiográfco según la medida del saco, con <2.5 mm como el límite para sacos foliculares normales. Dicho diagnóstico fue comparado con el respectivo diagnóstico histopatológico. Se calculó sensibilidad y especifcidad; se aplicó la prueba de chi-cuadrado, exacta de Fisher y, fnalmente, el índice Kappa. Resultados: se obtuvo un alto grado de acuerdo entre los observadores. La prueba radiográfca tuvo una baja sensibilidad (0.27) y especifcidad (0.6) y no se encontró diferencia estadísticamente signifcativa entre estos. Conclusiones: la ausencia de hallazgos radiográfcos no implica ausencia de enfermedad. Además, no se puede establecer relación entre la presencia de quistes dentígeros y radiolucidez ≥ 2.5 mm en radiografía panorámica digital.

Objective: To establish the relationship between the histopathological diagnosis of follicular sacs of third molars and the standardized radiographic measurement in digital panoramic radiography. Methods: This was a descriptive study in which 28 follicular sacs of third molars were included. In digital panoramic radiographs two observers measured the pericoronal radiolucency using a standardized method and the intraclass correlation index was calculated. A radiographic diagnosis was established according to the size of the sac, with <2.5mm being the limit for normal follicular sacs. This diagnosis was compared with the respective histopathological diagnosis. Sensitivity and specifcity were calculated; the chi-square test, Fisher's exact test and fnally the Kappa index were applied. Results: A high degree of agreement was obtained among the observers. The radiographic test had a low sensitivity (0.27) and specifcity (0.6) and no statistically signifcant diference was found between these. Conclusions: The absence of radiographic fndings does not imply absence of disease, furthermore, no relationship can be established between the presence of dentigerous cysts and radiolucency ≥ 2.5 mm in digital panoramic radiography.

Revisiting the human dental follicle: From tooth development to its association with unerupted or impacted teeth and pathological changes.

Dental follicles are involved in odontogenesis, periodontogenesis, and tooth eruption. Dental follicles are unique structures, considering that their remnants can persist within the jawbones after odontogenesis throughout life if the tooth does not erupt. Pathological changes may occur in these tissues as individuals age. The changes range from benign to life threatening. Thus, the assessment of age-related changes in dental follicles associated with unerupted teeth is of paramount importance. In this review, we summarize the physiological roles and changes in dental follicles in odontogenesis, tooth eruption, and aging, in addition to the pathological changes associated with these structures. We encourage investigators to consider this peculiar tissue as a unique model and explore its potential to clarify its importance from the viewpoints of developmental biology, tissue physiology, and pathology.

Age-Related Metabolic Pathways Changes in Dental Follicles: A Pilot Study.

Aging is not a matter of choice; it is our fate. The "time-dependent functional decline that affects most living organisms" is coupled with several alterations in cellular processes, such as cell senescence, epigenetic alterations, genomic instability, stem cell exhaustion, among others. Age-related morphological changes in dental follicles have been investigated for decades, mainly motivated by the fact that cysts and tumors may arise in association with unerupted and/or impacted teeth. The more we understand the physiology of dental follicles, the more we are able to contextualize biological events that can be associated with the occurrence of odontogenic lesions, whose incidence increases with age. Thus, our objective was to assess age-related changes in metabolic pathways of dental follicles associated with unerupted/impacted mandibular third molars from young and adult individuals. For this purpose, a convenience sample of formalin-fixed paraffin-embedded (FFPE) dental follicles from young (<16 y.o., n = 13) and adult (>26 y.o., n = 7) individuals was selected. Samples were analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS)-based untargeted metabolomics. Multivariate and univariate analyses were conducted, and the prediction of altered pathways was performed by mummichog and Gene Set Enrichment Analysis (GSEA) approaches. Dental follicles from young and older individuals showed differences in pathways related to C21-steroid hormone biosynthesis, bile acid biosynthesis, galactose metabolism, androgen and estrogen biosynthesis, starch and sucrose metabolism, and lipoate metabolism. We conclude that metabolic pathways differences related to aging were observed between dental follicles from young and adult individuals. Our findings support that similar to other human tissues, dental follicles associated with unerupted tooth show alterations at a metabolic level with aging, which can pave the way for further studies on oral pathology, oral biology, and physiology.

Effects of bleomycin on tooth eruption: a novel potential application.

There are many kinds of potentially undesirable teeth. At present, surgical extraction is the most efficient way to eliminate these teeth, but it's very complex and invasive. In this study, we investigated the effects of bleomycin (BLM) on dental follicle and tooth eruption as a potential conservative therapy for undesirable teeth. Our data showed that local injection of 0.2 U/kg BLM had no significant effects on tooth eruption compared to the control group in Wistar rats. With higher dose of BLM (0.5 or 2 U/kg), the eruption of treated teeth was interrupted and their root formation failed until 4 weeks postnatal without significant systemic toxicity. Additionally, those effects were not depending on the toxicity of overdose evidenced by TUNEL assay. In summary, injecting BLM into dental follicle at an early stage could interrupt tooth development and eruption, and may prevent the potentially clinical problems resulting from undesirable teeth instead of surgical removal.

Hyperplastic dental follicle: case report / Folículo pericoronário hiperplásico: relato de caso

ABSTRACT The dental follicle is characterized as a radiolucent area around an impacted tooth. It is believed that these follicles can originate cysts and tumors. The aim of this study was to report a clinical case of a 15-year-old female, normosystemic patient, with an increased dental follicle involving the crown of the tooth 48, which was removed and referred for histopathological examination. The diagnosis hypothesis was dentigerous cyst, but the histopathological report describes a definitive diagnosis of dental follicle. After six months of postoperative follow-up the patient presented no signs of relapse or complaints in the region.

RESUMEN El folículo pericoronario se caracteriza por un área radiolúcida alrededor de un diente no erupcionado. Se cree que eses folículos puedan desarrollar quistes y tumores. El objetivo de este trabajo es reportar el caso clínico de un paciente de género femenino, de 15 años de edad, buen estado general, que presentaba un folículo dilatado, cubriendo la corona del diente 48, lo cual ha sido removido y enviado para estudio histopatológico. La hipótesis diagnóstica fue de quiste dentígero, pero el informe histopatológico logró establecer como diagnóstico definitivo folículo pericoronario. En los primeros seis meses del período postoperatorio, no hay señales de recidiva o quejas en la región.

RESUMO O folículo pericoronário caracteriza-se como uma área radiolúcida em volta de um dente incluso. Acredita-se que esses folículos podem originar cistos e tumores. O objetivo deste trabalho foi relatar o caso clínico de um paciente do gênero feminino, 15 anos de idade, normossistêmica, que apresentava folículo pericoronário aumentado, envolvendo a coroa do dente 48, o qual foi removido e encaminhado para exame histopatológico. A hipótese diagnóstica foi cisto dentígero, mas o laudo histopatológico teve como diagnóstico definitivo folículo pericoronário. A paciente encontra-se com seis meses de pós-operatório sem sinais de recidiva ou queixas na região.

Low-intensity pulsed ultrasound promotes tissue regeneration in rat dental follicle cells in a porous ceramic scaffold

Abstract The aim of this study was to investigate the effects of low-intensity pulsed ultrasound (LIPUS) on the osteogenic differentiation of dental follicle cells (DFCs) in vitro and on the regenerative effects of DFC-OsteoBoneTM complexes in vivo. DFCs were isolated and characterized. In the in vitro study, DFCs were cultured in an osteogenic medium in the presence or absence of LIPUS. The expression levels of ALP, Runx2, OSX, and COL-I mRNA were analyzed using real-time polymerase chain reaction (RT-PCR) on day 7. Alizarin red staining was performed on day 21. The state of the growth of the DFCs that were seeded on the scaffold at 3, 5, 7, and 9 days was detected by using a scanning electron microscope. In our in vivo study, 9 healthy nude mice randomly underwent subcutaneous transplantation surgery in one of three groups: group A, empty scaffold; group B, DFCs + scaffold; and group C, DFCs + scaffold + LIPUS. After 8 weeks of implantation, a histological analysis was performed by HE and Mason staining. Our results indicate that LIPUS promotes the osteogenic differentiation of DFCs by increasing the expression of the ALP, Runx2, OSX, and COL-I genes and the formation of mineralized nodules. The cells can adhere and grow on the scaffolds and grow best at 9 days. The HE and Mason staining results showed that more cells, fibrous tissue and blood vessels could be observed in the DFCs + scaffold + LIPUS group than in the other groups. LIPUS could promote the osteogenic differentiation of DFCs in vitro and promote tissue regeneration in a DFCs-scaffold complex in vivo. Further studies should be conducted to explore the underlying mechanisms of LIPUS.

Study of Pathological Changes in the Dental Follicle of Disease-Free Impacted Third Molars.

BACKGROUND: The prophylactic extraction of third molars is a common practice in dental offices, but divergent opinions are found in the literature regarding the indication of this procedure. The aim of the present study was to determine the prevalence of pathological changes associated with the pericoronal tissue of asymptomatic impacted third molars that could justify prophylactic extraction. MATERIALS AND METHODS: A cross-sectional observational study was conducted in which 109 pericoronal tissues with no radiographic evidence of pathology were histopathologically analyzed. The specimens were fixed in 10% formalin, embedded in paraffin, stained with hematoxylin and eosin and analyzed individually by two pathologists. RESULT: The frequency of inflammatory infiltrate in the dental follicle of patients older than 20 years of age was significantly higher than that of younger patients (p = 0.004), demonstrating an association between inflammation in the dental follicle and patient age. The occurrence of squamous metaplasia was also greater in patients older than 20 years (p = 0.042), demonstrating that the prevalence of squamous metaplasia increases with age. A significant association was also found between inflammation and squamous metaplasia (p < 0.001). CONCLUSION: Pathological changes may be present in the dental follicle of impacted third molars even in the absence of clinical or radiographic signs of disease.

Aspectos radiográficos de imágenes periapicales asociadas a incisivos primarios traumatizados / Aspectos radiográficos de imagens periapicais associadas a incisivos decíduos traumatizados / Radiographic aspects of periapical images associated with traumatized primary incisors

Las imágenes radiolúcidas periapicales que involucran dientes primarios traumatizados se pueden confundir entre sí y dar lugar a diagnósticos erróneos y tratamiento. Por lo tanto, es importante identi-ficar las características radiográficas de las imágenes radiolúcidas periapicales en los incisivos prima-rios traumatizados, principalmente debido al hecho de que la superposición de imágenes se produce en esta región. Además, es frecuente observar la expansión del folículo haciendo que el diagnóstico radiográfico sea aún más difícil. El objetivo de este estudio fue describir, a través de una revisión de la literatura, las características radiográficas de las imágenes periapicales asociadas con los incisivos primarios traumatizados.

Periapical radiolucencies involving traumatized primary teeth can be confused with each other and lead to misdiagnosis and treatment. Therefore, it is important to identify radiographic characteristics of periapical radiolucent images in traumatized primary incisors, mainly due to the fact that overla-pping of images occurs in this region. Besides, it is frequent to observe follicle expansion making the radiographic diagnosis even more difficult. The objective of this study was to describe, through a literature review, the radiographic characteristics of periapical images associated with traumatized primary incisors.

As radioluscências periapicais envolvendo dentes decíduos traumatizados podem ser confundidas entre si e levar a um erro de diagnóstico e tratamento. Por isso, é importante identificar características radiográficas de imagens radiolúcidas periapicais em incisivos decíduos traumatizados, principalmente pelo fato que nesta região ocorre sobreposição de imagens e, é frequente observar expansão do folículo, dificultando ainda mais o diagnóstico radiográfico. O objetivo deste trabalho foi descrever através de uma revisão de literatura as características radiográficas de imagens periapicais associadas a incisivos decíduos traumatizados.

SCAPs Regulate Differentiation of DFSCs During Tooth Root Development in Swine.

The tooth root transmits and balances occlusal forces through the periodontium to the alveolar bone. The periodontium, including the gingiva, the periodontal ligament, the cementum and the partial alveolar bone, derives from the dental follicle (DF), except for the gingiva. In the early developmental stages, the DF surrounds the tooth germ as a sphere and functions to promote tooth eruption. However, the morphological dynamics and factors regulating the differentiation of the DF during root elongation remain largely unknown. Miniature pigs are regarded as a useful experimental animal for modeling in craniofacial research because they are similar to humans with respect to dentition and mandible anatomy. In the present study, we used the third deciduous incisor of miniature pig as the model to investigate the factors influencing DF differentiation during root development. We found that the DF was shaped like a crescent and was located between the root apical and the alveolar bone. The expression levels of WNT5a, ß-Catenin, and COL-I gradually increased from the center of the DF (beneath the apical foramen) to the lateral coronal corner, where the DF differentiates into the periodontium. To determine the potential regulatory role of the apical papilla on DF cell differentiation, we co-cultured dental follicle stem cells (DFSCs) with stem cells of the apical papilla (SCAPs). The osteogenesis and fibrogenesis abilities of DFSCs were inhibited when being co-cultured with SCAPs, suggesting that the fate of the DF can be regulated by signals from the apical papilla. The apical papilla may sustain the undifferentiated status of DFSCs before root development finishes. These data yield insight into the interaction between the root apex and surrounding DF tissues in root and periodontium development and shed light on the future study of root regeneration in large mammals.

Impactação de dente decíduo ­ relato de caso / Deciduous tooth impaction ­ case report

Introdução: A impactação dentária raramente ocorre na dentição decídua, e quando ocorre, pode ocasionar alterações funcionais e riscos de lesões patológicas. Objetivo: Relatar um caso raro de impactação dentária na dentição decídua e a conduta terapêutica realizada. Relato de caso: Paciente, sexo feminino, 9 anos de idade, procurou atendimento devido à ausência de dente na região anterior. No exame clínico e radiográfico foi constatado agenesia do dente 12 e impactação do elemento 52 associado à imagem sugestiva de lesão envolvendo o elemento dentário. A paciente foi submetida à remoção cirúrgica do elemento e da lesão. O material coletado na cirurgia foi encaminhado para exame histopatológico, sendo diagnóstico como Capuz Hiperplásico. Em seguida, a paciente foi encaminhada para o tratamento reabilitador. Conclusão: A remoção cirúrgica do dente impactado e da lesão, associada ao diagnóstico radiográfico e histopatológico apropriados, foram eficazes para o caso clinico apresentado (AU).

Introduction: Primary tooth impaction is a rare phenomenon in deciduous dentition and when it occurs, it may lead to functional alterations and risks of pathological lesions. Objective: To report a rare case of dental impaction in deciduous dentition and the therapeutic approach taken. Case report: Patient, female, 9 years old sought dental assistance due to the absence of teeth in the anterior maxillary region. The clinical and radiographic examinations identified absence of tooth 12 and impaction of tooth 52 associated to image suggesting lesion involving the teeth. A surgical intervention under local anesthesia was done for the removal of the tooth and the lesion. The material collected in the surgery was sent for histopathological examination and diagnosed as dental follicles. Then the patient was referred for rehabilitation treatment. Conclusion: The surgical removal of impacted tooth and of lesion associated with appropriate radiographic and histopathology diagnosis were effective in the clinical case presented (AU).

Dental Follicle Cells Participate in Tooth Eruption via the RUNX2-MiR-31-SATB2 Loop.

Cleidocranial dysplasia (CCD) is characterized by the runt-related transcription factor 2 (RUNX2) mutation, which results in delayed tooth eruption due to disturbed functions of dental follicle. Accumulating evidence has revealed a key regulatory circuit, including RUNX2, miR-31, and special AT-rich binding protein 2 (SATB2) acting in concert in mesenchymal stem cell homeostasis and functions. However, whether such a regulatory loop works in dental follicle cells (DFCs) remains unknown. Herein, we investigated the roles of RUNX2-miR-31-SATB2 in DFCs from patients with CCD (DFCs-CCD) to advance our understanding regarding physical tooth eruption. We identified a novel mutation on exon 5 (c.634T>G, p.T212P) in RUNX2 via exome sequencing in the CCD patient with typical clinical presentations. Compared with DFCs from healthy donors, DFCs-CCD displayed significantly lower osteogenic, osteoclast-inductive, and matrix-degrading capacities and had lower RUNX2 (a transcriptional inhibitor of miR-31), higher miR-31, and downregulated SATB2. Lower ratios of RANKL/OPG and RANKL/RANK, as well as decreased expression of matrix metalloproteinase 9 (MMP9) and matrix metalloproteinase 2 (MMP2), would lead to inactivation of osteoclasts and suppression of bone matrix remodeling in DFCs-CCD. Furthermore, the roles of the RUNX2-miR-31-SATB2 loop in DFCs-CCD were revealed by endogenous miR-31 knockdown, which resulted in increased SATB2 and RUNX2, as well as osteoclast-inductive and matrix degradation capacities. Conversely, SATB2, RUNX2, MMP9, MMP2, and osteoclast-inductive factors expression declined upon ectopic miR-31 overexpression in normal DFCs. Importantly, neonatal mice with in vivo siRUNX2 delivery exhibited less activated osteoclasts around dental follicles and delayed tooth eruption. Together, these results suggest that RUNX2 mutation/haploinsufficiency disturbs osteoclast-inductive signaling in DFCs, which may be responsible for delayed tooth eruption in CCD patients. Manipulation of the RUNX2-miR-31-SATB2 loop may be a potential way to facilitate tooth eruption in CCD patients.

Incidência de anormalidades histológicas em tecido correspondente ao espaço pericoronário de terceiros molares inclusos e semi-inclusos / Incidence of the soft tissue changes around unerupted or impacted third molars

Introdução: O Folículo Pericoronário envolve a coroa do germe dental durante seu desenvolvimento. Quando o dente permanece incluso, alterações do folículo podem originar doenças, como cistos e tumores odontogênicos. Objetivo: Analisar as alterações histológicas no tecido mole circundante a terceiros molares inclusos e semi-inclusos, independentemente de alterações patológicas aparentes em suas radiografias correspondentes, além de relacionar o diagnóstico histológico com o diagnóstico radiográfico dos casos. Material e Método: A partir de terceiros molares extraídos de 26 pacientes, foram analisados espécimes histológicos de folículos pericoronários por dois examinadores calibrados. O diagnóstico histopatológico obtido foi relacionado ao radiográfico, sendo este realizado por meio de radiografias periapicais e, quando necessário, complementado por radiografias panorâmicas. Resultado: Dos 37 folículos pericoronários avaliados, 30% mostraram alterações histológicas compatíveis com cistos dentígeros; 51% eram folículos normais, e 19% continham apenas fragmentos de mucosa. Radiograficamente, 100% dos casos demonstraram características de folículos sem alterações. Conclusão: Esses dados exemplificam que anormalidades podem estar presentes nos tecidos pericoronários sem que haja evidências clínicas e radiográficas. O exame histopatológico provê um diagnóstico mais preciso e deve ser considerado para a construção do diagnóstico definitivo. .

Introduction: Dental follicles remain adjacent to the crown of an unerupted or impacted tooth. Changes in dental follicles can occur and form a solitary cyst around the crown of the tooth, deemed dentigerous cysts. Objective: The aim of this study was to determine the incidence of soft tissue changes around unerupted or impacted third molars. Material and Method: Pericoronal tissue of patients who were referred to our clinic for removal of third molars for a variety of reasons was examined histopathologically. Follicular spaces were measured from periapical and panoramic radiographs by each author independently. Result: Of the 37 specimens submitted for histopathologic examination, 19 normal follicular tissues (51%), 11 arrangements typical of a cyst (30%) and 7 normal tissues of oral mucous (11%) were found. Conclusion: Cystic changes may be encountered in the histopathologic examination of radiographically normal unerupted or impacted third molar follicles. .

Bone morphogenetic protein 9-induced osteogenic differentiation of dental follicle cells in vitro / 中国组织工程研究

BACKGROUND:Bone morphogenetic protein 9 is proved to promote the osteogenic differentiation of various kinds of stem cel s, but whether it can induce the osteogenic differentiation of dental fol icle cel s in vitro is yet unclear. OBJECTIVE:To investigate whether bone morphogenetic protein 9 can induce the osteogenic differentiation of rat dental fol icle cel s in vitro. METHODS:Purified rat dental fol icle cel s at passage 3 were transfected with bone morphogenetic protein 9 adenovirus. Then, alkaline phosphatase activity, calcium deposition and expression of osteogenesis-related factors at mRNA and protein levels were detected in the dental fol icle cel s. RESULTS AND CONCLUSION:After transfection with bone morphogenetic protein 9, the dental fol icle cel s showed continuously enhanced alkaline phosphatase activities and obviously enhanced calcium deposition. Real-time PCR results demonstrated that the mRNA expressions of alkaline phosphatase, osteocalcin, bone sialoprotein, osteopontin and core binding factor were increased significantly. The western blot assay showed that the expression of osteopontin enhanced in the dental fol icle cel s after transfection with bone morphogenetic protein 9. In summary, bone morphogenetic protein 9 can induce the osteogenic differentiation of dental fol icle cel s.

Expression of osteoblast-specific genes in immortalized rat dental follicle cells / 中国组织工程研究

BACKGROUND:During in vitro culture, dental folicle cels are easy to loss self-renewal capacity and become aging, and they are also very difficult to be purified and amplified in quantity, which limits the application of dental folicle cels in periodontal tissue engineering. OBJECTIVE:To study the osteogenic effect of immortalized rat dental folicle cels. METHODS:pSSR69-pAmpho plasmids containing SV40T-Ag were used to transfect 293 cels. Rat dental folicle cels were transfected with virus supernatant and screened by hygromycin to establish immortalized rat dental folicle cels (experimental group). Untransfected cels served as controls. RT-PCR was used to detect the osteogenic related factors (alkaline phosphatase, osteocalcin, bone morphogenetic protein 2, and Runx2) in the experimental group and control group. RESULTS AND CONCLUSION:The results showed no statistical differences between the experimental group and the control group in the expressions of alkaline phosphatase, bone morphogenetic protein 2 and Runx2 (P > 0.05). The expression of osteocalcin was significantly higher in the experimental group than the control group (P < 0.05). These findings suggest that in the late osteogenesis differentiation, immortalized rat dental folicle cels may promote the secretion of osteocalcin and then make osteoblasts early entry into the bone calcification stage.

Do dental stem cells depict distinct characteristics? - Establishing their "phenotypic fingerprint".

Dental tissues provide an alternate source of stem cells compared with bone marrow and have a similar potency as that of bone marrow derived mesenchymal stem cells. It has been established there are six types of dental stem cells: Dental pulp stem cells, stem cells from human exfoliated deciduous teeth, stem cells from apical papilla, periodontal ligament stem cells, dental follicle progenitor cells, oral periosteum stem cells and recently gingival connective tissue stem cells. Most of the dental tissues have a common developmental pathway; thus, it is relevant to understand whether stem cells derived from these closely related tissues are programmed differently. The present review analyzes whether stem cells form dental tissues depict distinct characteristics by gaining insight into differences in their immunophenotype. In addition, to explore the possibility of establishing a unique phenotypic fingerprint of these stem cells by identifying the unique markers that can be used to isolate these stem cells. This, in future will help in developing better techniques and markers for identification and utilization of these stem cells for regenerative therapy.

Estudo das células primárias dentais, viabilidade celular e bioatividade de materiais á base de silicato de cálcio / Cell viability and bioactivity of Portland cement-based materials in human dental follicle and pulp stem cells, fibroblasts and osteoblasts

Os objetivos deste estudo foram o isolamento e diferenciação de células da polpa (hDPCs) e do folículo (hDFCs) dentário de terceiros molares humanos e utilização para avaliação da citotoxicidade e bioatividade de novos cimentos endodônticos. (Capítulo 1) - Após a obtenção das culturas primárias foram realizados os ensaios vermelho de Alizarina, atividade da enzima fosfatase alcalina (ALP). A expressão gênica de proteína morfogenética óssea (bmp7), sialofosfoproteína dentinária (dspp) e alp foi avaliada. Os ensaios de bioatividade demonstraram maior atividade para hDPCs, além de maior expressão de dspp e alp; a expressão de bmp7 foi semelhante nas duas linhagens. Conclui-se que hDPCs demonstram maior capacidade de diferenciação em células com potencial de mineralização, quando comparadas com hDFCs. (Capítulo 2) – Foram utilizadas linhagens primárias (hDPCs e hDFCs), em comparação com linhagens imortalizadas (células osteoblásticas humanas Saos-2, e do ligamento periodontal de ratos mPDL) para análise dos materiais: 1) Cimento Portland branco (CP); 2) MTA; 3) CP/ óxido de nióbio microparticulado (Nb2O5?); 4) CP/ óxido de nióbio nanoparticulado (Nb2O5n). A citotoxicidade foi avaliada pelos ensaios MTT e Azul de Trypan e a bioatividade pela atividade da ALP. Viabilidade celular foi observada em todas as linhagens; e a atividade de ALP apresentou maior resposta na linhagem Saos-2. Conclui-se que as diferentes linhagens celulares são similares para avaliação da citotoxicidade; entretanto, para avaliação da bioatividade o uso de uma linhagem celular osteoblástica é mais indicado. Além disso, observa-se que materiais à base de Nb2O5? e Nb2O5n mostram potencial como radiopacificador alternativo no MTA

The aims of this study were to isolate and differentiate pulp (hDPCs) and follicle (hDFCs) cells obtained from human third molars and to use them in cytotoxicity and bioactivity assays of new dental materials. (Chapter 1) - After obtaining the stem cultures, Alizarin Red, alkaline phosphatase (ALP) activity and gene expression by Real Time PCR of bone morphogenetic protein (BMP7), dentin sialophosphoprotein (DSPP) and ALP were performed. The bioactivity asssays showed greater activity for hDPCs, and increased expression of dspp and alp; the expression of bmp7 was similar in both cell lines. We conclude that hDPCs demonstrate greater ability to differentiate into cells with mineralization potential when compared with hDFCs. (Chapter 2) - stem lineages (hDPCs and hDFCs) were used, compared to immortalized lineages (human osteoblastic cells Saos-2, and periodontal ligament of mice cells mPDL) for analysis of the following materials: 1) white Portland cement (PC); 2) MTA; 3) C/ microparticulated of niobium oxide (Nb2O5µ); 4) CP/nanoparticulated niobium oxide (Nb2O5n). Cytotoxicity was performed by MTT and Trypan Blue assays and bioactivity by ALP activity assay. Cell viability was observed in all cell lines, and ALP activity showed greater response in Saos-2. We conclude that the different cell lines are similar in cytotoxicity, however, to evaluate the bioactivity, using an osteoblastic cell line is more appropriate. Moreover, it is observed that materials based on Nb2O5µ and Nb2O5n show potential for alternative use to MTA

PrPc is temporospatially expressed in molar development of rats.

Odontogenesis, tooth development, is derived from two tissue components: ectoderm and neural crest-derived mesenchyme. Cyto-differentiation of odontogenic cells during development involves time-dependent and sequential regulation of genetic programs. This study was conducted to detect molecules implicated in cyto-differentiation of developing molar germs of rats. Differential display-PCR revealed that PrP(c) was differentially expressed between cap/early bell-staged germs (maxillary 3rd molar germs) and root formation-staged germs (maxillary 2nd molar germs) at postnatal day 9. Both levels of PrP(c) mRNA and protein expression were higher in the root formation stage than the cap/early bell stage and increased in a time-dependent manner. Immunofluorescence revealed for the first time that PrP(c) was not localized in the enamel organ, but localized in dental follicular cells for the development of the periodontal ligament and cementum as well as odontoblasts, both of which are of neural crest origin. These results suggest that the physiological functions of the PrP(c) in tooth development may be implicated in the differentiation of neural crest-derived mesenchyme including the periodontal tissues for root formation rather than epithelial tissue.

Immunohistochemical Analysis of Proliferating Cell Nuclear Antigen (PCNA) in Dental Follicles of Impacted Third Molars / Análisis Imunohistoquímico de Antígeno de Proliferación Celular Nuclear (PCNA) en Folículos Dentales de los Terceros Molares Retenidos

This study investigated the immunodetection of PCNA in epithelial components of dental follicles associated with impacted third molars without radiographical and morphological signs of pathosis. A total of 105 specimens of dental follicles associated with impacted third molars with incomplete rhizogenesis (between Nolla's stage 6 and 9) were surgically removed from 56 patients. Epithelial cell proliferating was determined by using immunohistochemical labeling. Statistical analysis was performed using the Fisher exact test. Of the 105 dental follicles collected, 6 were PCNA-positive ( 6 percent). The specimens with squamous metaplasia and epithelial hyperplasia had higher rates of positivity for PCNA, as well as those with proliferative remnants of odontogenic epithelium. In conclusion, this study shows that dental follicles at this stage of development have low proliferative potential, but suggests that squamous metaplasia, hyperplasia of the epithelial lining and presence of proliferative odontogenic epithelial rests in the connective tissue may be early signs of developing lesions of odontogenic origin.

Se investigó la inmunodetección de PCNA en los componentes epiteliales de los folículos dentales asociados a terceros molares retenidos sin signos radiográficos y morfológicos de la patología. Fueron extraídos quirúrgicamente, de 56 pacientes, 105 muestras de folículos dentales asociados a terceros molares retenidos con rizogénesis incompleta (entre los estadíos de Nolla 6 y 9) La proliferación de células epiteliales se deteminó mediante inmunohistoquímica. El análisis estadístico se realizó mediante la prueba exacta de Fisher. De los 105 folículos dentales recogidos, 6 fueron PCNA-positivos ( 6 por ciento). Las muestras con metaplasia escamosa e hiperplasia epitelial tuvieron mayores tasas de positividad para PCNA, así como aquellos con restos de proliferación del epitelio odontogénico. En conclusión, este estudio mostró que los folículos dentales en esta etapa del desarrollo tienen un potencial proliferativo bajo, pero sugiere que la metaplasia escamosa, la hiperplasia del epitelio y la presencia de restos epiteliales odontogénicos, en proliferación en el tejido conectivo, pueden ser signos tempranos de lesiones en el desarrollo de origen odontogénico.

Expressão da podoplanina em ameloblastomas e folículos pericoronários de dentes não-irrompidos: estudo comparativo com a expressão do Ki-67 / Podoplanin expression in ameloblastomas and dental follicles of unerupted teeth: comparative estudy with Ki-67 expression

A relação da podoplanina com a proliferação celular tem sido demonstrada, porém ainda é tema de controvérsia. O objetivo deste estudo foi investigar a expressão da podoplanina em ameloblastomas e remanescentes de epitélio odontogênico (REO) de folículos pericoronários de dentes não-irrompidos e verificar a relação da imunomarcação da podoplanina com a proliferação celular. A amostra incluiu 33 pacientes submetidos à biópsia incisional ou à exérese de ameloblastomas e 32 folículos de pacientes submetidos à extração de dentes não-irrompidos. Todos os espécimes selecionados foram corados pela técnica imuno-histoquímica com os anticorpos anti-podoplanina e anti-Ki-67. A expressão da podoplanina foi avaliada por um método semi-quantitativo de escores. Já o índice de proliferação celular foi obtido pela porcentagem de células positivas (no mínimo 500 células/ameloblastoma e todas as células do REO foram avaliadas). Todos os ameloblastomas apresentaram imunomarcação pela podoplanina nas células periféricas. Nas células centrais, a expressão foi ausente ou fraca. Nos REO, a podoplanina apresentou expressão predominantemente nas células em contato com o tecido conjuntivo. A expressão membranosa da podoplanina foi superior nos ameloblastomas em relação aos REO (!=0,001). Também houve diferença significante entre a imunomarcação citoplasmática e membranosa da podoplanina nos REO (!=0,001). A expressão do Ki-67 foi mais acentuada nos ameloblastomas do que nos REO (!<0,001). A correlação entre a expressão citoplasmática (r= -0,15, != 0, 396) e membranosa (r= 0,00, != 0, 989) da podoplanina e do Ki-67 nos ameloblastomas não foi estatisticamente significante, tampouco entre a imunomarcação citoplasmática (r= 0,15, != 0, 421) e membranosa (r= -0, 09, ! = 0, 629) da podoplanina e do Ki-67 nos REO. Os resultados sugerem que não há associação da podoplanina com a proliferação celular e reforçam a necessidade da elucidação do papel desta proteína em tumores odontogênicos benignos.

The association between podoplanin and cellular proliferative activity has been demonstrated but is still source of debate. The aim of this study was to investigate the expression of podoplanin in ameloblastomas and remnants of odontogenic epithelium (ROE) from dental follicles of unerupted teeth and verify the relation between the podoplanin expression and proliferative activity of the odontogenic cells. Thirty-three patients submitted to the incisional biopsy or resection of ameloblastoma and 32 patients submitted to the extraction of unerupted tooth were selected for analysis. Archived paraffin-embedded ameloblastomas and dental follicles specimens were sectioned and stained with anti-human podoplanin and anti-Ki-67 antibodies. The podoplanin expression by odontogenic epithelial cells was evaluated using a score method and the Ki-67 labelling index was obtained by the percentage of positive odontogenic cells (at least 500 cells/ameloblastoma and all ROE cells). All ameloblastomas showed podoplanin expression in ameloblast-like cells of the epithelial islands. Weak or absent immunostaining was observed in the central cells resembling stellate reticulum. Podoplanin expression in ROE was mainly found in the cells in contact with the connective tissue. Membranous expression of podoplanin in ameloblastomas was stronger than its expression in ROE (!=0.001). Statistically significant difference between cytoplasmic and membranous expression of podoplanin in ROE was observed (!=0.001). The index of cellular odontogenic proliferative activity, verified by Ki-67 expression, was highest in ameloblastomas when compared with ROES (!<0.001). No statistically significant correlation between podoplanin and cellular odontogenic proliferative activity in ameloblastomas and dental follicles was found (!>0.05). These results support the evidence that there is no connection between podoplanin expression and odontogenic cellular proliferative activity in ameloblastomas and...

Correlation of the radiographic and morphological features of the dental follicle of third molars with incomplete root formation.

The objective of this study was to determine the correlation of the radiographic and morphological features of the dental follicle of unerupted third molars with incomplete root formation. A cross-sectional study was carried out with 56 patients (105 teeth) aged 13 to 24 years. Panoramic radiography was used to determine the stage of root formation to locate and measure pericoronal radiolucency. The width of the dental follicle ranged from 0.0 to 4.0 mm, the distal face being the one most frequently involved, and stage 7 of root formation showing the highest incidence. An inactive enamel reduced epithelium and inactive epithelium remnant also showed a high incidence. Dense connective tissue showed a high incidence, chronic inflammation was infrequent and calcification was a common finding. There was a significant association between the progression of the rhizogenesis and the transformation of the enamel reduced epithelium into a stratified squamous epithelium. No significant association was found between rhizogenesis and the other morphological findings or between the latter and the width of the pericoronal space. It was concluded that there was no clinically significant correlation between the radiographic and morphological features. Every asymptomatic unerupted third molar should be followed up and the follicular tissue analyzed.