RESUMEN
Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is limited in hypoxia, and HIF-1α is key to the response to hypoxia. However, its mechanisms remain largely unknown. This study discovered an osteogenesis-related gene sensitive to hypoxia in PDLSCs, and investigated the molecular mechanisms between HIF-1α and the gene. NOG, a gene that negatively regulates osteogenesis, was discovered by RNA-seq. Under normoxic conditions, HIF-1α overexpression led to enhanced expression of NOG/Noggin and inhibited the expression of osteogenesis-related genes, while inhibition of HIF-1α reversed this effect. The expression of HIF-1α, NOG/Noggin and the osteogenesis-related genes were detected by qRT-PCR or Western blot. Mechanistically, we verified that HIF-1α binds to the hypoxia response element (-1505 to -1502) in the promotor of NOG to enhance secretion of Noggin by chromatin immunoprecipitation and a dual-luciferase reporter assay. IHC staining findings in an animal model verified that Noggin-associated osteogenic differentiation was inhibited in hypoxia. NOG displayed a concordant relationship with HIF-1α, and secreted more with increasing of HIF-1α. Hypoxia stabilized HIF-1α, which bound to the HRE (-1505 to -1502) of the NOG promotor to enhance NOG transcription resulted in inhibiting osteogenic differentiation of PDLSCs. This study offers a promising therapy for periodontitis.
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Osteogénesis , Ligamento Periodontal , Animales , Diferenciación Celular/genética , Células Cultivadas , Hipoxia/metabolismo , Osteogénesis/genética , Ligamento Periodontal/metabolismo , Células MadreRESUMEN
Objective @# To investigate the application of digital immediate implant and angle screw channel abutment in the aesthetic area and the related influencing factors by reviewing the data of one case of immediate implant repair of the upper anterior teeth and related literature. @*Methods@#One case of refractory chronic apicitis of the upper anterior teeth involved immediate implantation after extraction. The digital information of the patient was obtained by CBCT and intraoral scanning. According to the information from the patients, a preoperative evaluation was performed; a treatment scheme was formulated; a minimally invasive extraction was performed; implants were placed under a digital guide plate; and temporary restoration was immediately performed. Six months after the operation, the patients underwent individualized mold removal, and angle screw channel fixation was completed. We observed the cosmetic effects and soft and hard tissue and gingival contour maintenance effects after restoration and reexamined the patients 6 months after restoration. In addition, the relevant literature was reviewed. @*Results @#The height of the gingival margin and gingival papilla and gingival contour of this patient were well maintained. The red and white aesthetic effect was good. There was no redness or swelling of the gingiva nor obvious changes in the soft and hard tissues around the implant 6 months after restoration, and the patient was satisfied. The results in the literature review show that a preoperative design based on CBCT and intraoral scanning data combined with digital software and a whole digital guide plate make the procedure more accurate and safer. These factors can not only avoid important anatomical structures and serious surgical complications but can also result in implantation in the best three-dimensional position. In addition, the application of digital impression technology and CAD/CAM increases the efficiency, speed, accuracy, simplicity, and comfort of oral impressions and the construction of temporary and final prostheses more precise and faster, greatly improving clinical efficiency. @*Conclusion@#Digital immediate implant and angle screw channel abutment is a good method to restore the aesthetics and function of missing teeth and to avoid the complications caused by adhesive residue.
RESUMEN
Periodontitis is characterized by the loss of periodontal tissues, especially alveolar bone. Common therapies cannot satisfactorily recover lost alveolar bone. Periodontal ligament stem cells (PDLSCs) possess the capacity of self-renewal and multilineage differentiation and are likely to recover lost alveolar bone. In addition, periodontitis is accompanied by hypoxia, and hypoxia-inducible factor-1α (HIF-1α) is a master transcription factor in the response to hypoxia. Thus, we aimed to ascertain how hypoxia affects runt-related transcription factor 2 (RUNX2), a key osteogenic marker, in the osteogenesis of PDLSCs. In this study, we found that hypoxia enhanced the protein expression of HIF-1α, vascular endothelial growth factor (VEGF), and RUNX2 ex vivo and in situ. VEGF is a target gene of HIF-1α, and the increased expression of VEGF and RUNX2 proteins was enhanced by cobalt chloride (CoCl2, 100 µmol/L), an agonist of HIF-1α, and suppressed by 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1, 10 µmol/L), an antagonist of HIF-1α. In addition, VEGF could regulate the expression of RUNX2, as RUNX2 expression was enhanced by human VEGF (hVEGF165) and suppressed by VEGF siRNA. In addition, knocking down VEGF could decrease the expression of osteogenesis-related genes, i.e., RUNX2, alkaline phosphatase (ALP), and type I collagen (COL1), and hypoxia could enhance the expression of ALP, COL1, and osteocalcin (OCN) in the early stage of osteogenesis of PDLSCs. Taken together, our results showed that hypoxia could mediate the expression of RUNX2 in PDLSCs via HIF-1α-induced VEGF and play a positive role in the early stage of osteogenesis of PDLSCs.
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Diferenciación Celular/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Regulación de la Expresión Génica , Ligamento Periodontal/citología , Células Madre/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Fosfatasa Alcalina/genética , Fosfatasa Alcalina/metabolismo , Hipoxia de la Célula , Células Cultivadas , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogénesis/genética , Interferencia de ARN , Células Madre/citología , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Periodontal ligament stem cells (PDLSCs) exhibit potential for osteogenesis in vitro and in vivo and are a candidate cell type for periodontal regeneration for the treatment of periodontitis. However, periodontitis is accompanied by hypoxia, and it is not clear how hypoxia affects the osteogenesis of PDLSCs. In this study, we found that the expression of hypoxia-inducible factor-1α (HIF-1α) and transforming growth factor-ß1 (TGF-ß1) is enhanced in the osteogenesis of PDLSCs under nonhypoxic conditions. TGF-ß1 can induce the stabilization of HIF-1α through the phosphorylation of mothers against decapentaplegic homolog 3 (Smad3) in PDLSCs, and in turn, HIF-1α inhibits the mRNA and protein expression of TGF-ß1 and inhibits the phosphorylation of Smad3 in PDLSCs. In addition, both HIF-1α and TGF-ß1 reduce the expression of crucial osteogenic gene runt-related transcription factor 2 (RUNX2) and the mineralization of PDLSCs in normoxia. In conclusion, our results showed that TGF-ß1 can induce the stabilization of HIF-1α in PDLSCs under nonhypoxic conditions, that HIF-1α can negatively regulate the TGF-ß1/Smad3 signal pathway in PDLSCs, and that TGF-ß1 and HIF-1α can synergistically inhibit the osteogenesis of PDLSCs.
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Subunidad alfa del Factor 1 Inducible por Hipoxia/fisiología , Osteogénesis , Ligamento Periodontal/citología , Células Madre/fisiología , Factor de Crecimiento Transformador beta1/fisiología , Adolescente , Diferenciación Celular , Hipoxia de la Célula , Células Cultivadas , Niño , HumanosRESUMEN
Globally, oral cancer is the most common type of head and neck cancers. Melatonin elicits inhibitory effects on oral cancer; however, the biological function of melatonin and underlying mechanisms remain largely unknown. In this study, we found that melatonin impaired the proliferation and apoptosis resistance of oral cancer cells by inactivating ROS-dependent Akt signaling, involving in downregulation of cyclin D1, PCNA, and Bcl-2 and upregulation of Bax. Melatonin inhibited the migration and invasion of oral cancer cells by repressing ROS-activated Akt signaling, implicating with the reduction of Snail and Vimentin and the enhancement of E-cadherin. Moreover, melatonin hampered vasculogenic mimicry of oral cancer cells through blockage of ROS-activated extracellular-regulated protein kinases (ERKs) and Akt pathways involving the hypoxia-inducible factor 1α. Consistently, melatonin retarded tumorigenesis of oral cancer in vivo. Overall, these findings indicated that melatonin exerts antisurvival, antimotility, and antiangiogenesis effects on oral cancer partly by suppressing ROS-reliant Akt or ERK signaling.
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Melatonina/uso terapéutico , Neoplasias de la Boca/tratamiento farmacológico , Especies Reactivas de Oxígeno/antagonistas & inhibidores , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Transición Epitelial-Mesenquimal , Humanos , Melatonina/farmacología , Neoplasias de la Boca/patología , Transducción de SeñalRESUMEN
OBJECTIVES: This study evaluated the accuracy of cone-beam computed tomography (CBCT) in detecting the root canal morphology of mandibular first premolars using micro-computed tomography (micro-CT) as a reference standard. MATERIALS AND METHODS: In total, 143 extracted human mandibular first premolars were selected and scanned using micro-CT and CBCT. The acquired images were used to evaluate the root canal morphology in each tooth, and evaluations were repeated after 2 weeks. The root canal configurations observed on the three-dimensional images were recorded, and the findings from both modalities were compared using chi-square tests. The actual agreement between the two modalities was assessed using kappa statistics. RESULTS: In total, the root morphologies in 136 mandibular first premolars were consistently identified by both CBCT and micro-CT: type I in 104, type III in five, type V in 20, and type IX in seven. Of the remaining seven teeth, the morphology in two, one, and four teeth was identified as type I, type VII, and type IX (type 1-3 in two and type 1-2-3 in two), respectively, by micro-CT and misdiagnosed as type III, type V, and type V, respectively, by CBCT. There were no significant differences between the two modalities with regard to the accurate detection of root canal configurations, with a kappa value of 0.886 for the actual agreement. CONCLUSIONS: Although CBCT may be accurate in detecting the root canal configuration in mandibular first premolars, it produces poorer image details compared with micro-CT. CLINICAL RELEVANCE: CBCT is a reliable radiological technique, but its accuracy in detecting details of the root canal morphology in mandibular first premolars, especially in some complex root canal configurations, needs to be improved.
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Diente Premolar/anatomía & histología , Diente Premolar/diagnóstico por imagen , Tomografía Computarizada de Haz Cónico , Cavidad Pulpar/anatomía & histología , Cavidad Pulpar/diagnóstico por imagen , Microtomografía por Rayos X , Humanos , Técnicas In Vitro , Mandíbula/anatomía & histología , Mandíbula/diagnóstico por imagenRESUMEN
Accumulating evidences have demonstrated that lipopolysaccharide (LPS) represents the important etiologic factor for sepsis. Some previous studies have reported the relationship between common polymorphisms rs4986790 and rs4986791 in the coding gene for this receptor and the susceptibility to sepsis, but there were distinct divergences between those findings. We therefore designed this meta-analysis incorporated 28 published articles containing 6,537 sepsis patients and 8,832 controls for a more comprehensive conclusion on this matter. Odds ratios (ORs) and 95% confidence interval (95% CIs) were calculated to evaluate the association of toll like receptor 4 gene polymorphisms rs4986790 and rs4986791 with sepsis risk. Heterogeneity between included studies was inspected using Q test, and sensitivity analysis was implemented via sequential deletion of each included study to investigate the stability of overall estimates. Funnel plot and Egger's test were adopted to examine publication bias across selected studies. We found no significant association for either the polymorphism rs4986790 or rs4986791 with sepsis susceptibility in total analysis under any genetic models. Neither did we after combining these two polymorphisms. The results of this meta-analysis suggest that the rs4986790 and rs4986791 polymorphisms in toll like receptor 4 gene may have no statistically significant influence on sepsis susceptibility.
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Predisposición Genética a la Enfermedad , Polimorfismo Genético , Sepsis/genética , Receptor Toll-Like 4/genética , HumanosRESUMEN
Cranial neural crest-derived cells (CNCCs) play important role in epithelial-mesenchymal interactions during tooth morphogenesis. However, the heterogeneity of CNCCs and their tendency to spontaneously differentiate along smooth muscle or osteoblast lineages in vitro limit further understanding of their biological properties. We studied the differentiation properties of isolated rat embryonic postmigratory CNCCs, expressing p75 neurotrophin receptor (p75NTR). These p75NTR positive (p75(+)) CNCCs, isolated using fluorescence activated cell sorter, exhibited fibroblast-like morphology and characteristics of mesenchymal stem cells. Incubation of p75(+) CNCCs in dental follicle cell conditioned medium (DFCCM) combined with dentin non-collagenous proteins (dNCPs), altered their morphological features to cementoblast-like appearance. These cells also showed low proliferative activity, high ALP activity and significantly increased calcified nodule formation. Markers related to mineralization or specific to cementoblast lineage were highly expressed in dNCPs/DFCCM-treated p75(+) cells, suggesting their differentiation along cementoblast-like lineage. p75(+) stem cells selected from postmigratory CNCCs represent a pure stem cell population and could be used as a stem cell model for in vitro studies due to their intrinsic ability to differentiate to neuronal cells and transform from neuroectoderm to ectomesenchyme. They can provide a potential stem cell resource for tooth engineering studies and help to further investigate mechanisms of epithelial-mesenchymal interactions in tooth morphogenesis.
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Diferenciación Celular , Células-Madre Neurales/fisiología , Fosfatasa Alcalina/metabolismo , Animales , Calcificación Fisiológica , Movimiento Celular , Forma de la Célula , Células Cultivadas , Medios de Cultivo Condicionados , Cemento Dental/citología , Saco Dental/citología , Cresta Neural/citología , Ratas Sprague-DawleyRESUMEN
OBJECTIVES: This study aims to explore the correlation between radicular grooves and root canal types by quantitatively detecting the radicular groove of mandibular first premolars using micro-computed tomography. MATERIALS AND METHODS: A total of 127 mandibular first premolars were scanned by micro-computed tomography, and 52 teeth with radicular grooves were identified. Details of root canal type and groove length, depth, and location were analyzed from three-dimensional images. RESULTS: A total of 40.9 % (52/127) of teeth had radicular grooves. Most of the grooves (69.5 %) were located on the mesial surface of the root. The prevalence of radicular grooves in single canals (17.4 %; 15/86) was lower than that in multiple and complex canals (90.2 %; 37/41); this difference was statistically significant (P < 0.001). The mean length and depth of radicular groove in type V (7.7 ± 2.16 and 0.87 ± 0.39 mm, respectively) and other types of canals (6.91 ± 2.67 and 0.63 ± 0.27 mm, respectively) were significantly longer and deeper than type I canals (6.06 ± 2.12 and 0.43 ± 0.14 mm, respectively). CONCLUSIONS: Multiple and complex canals had a higher incidence of radicular grooves and more complicated root morphology than single and simple canals. Therefore, the anatomy of radicular grooves may influence root canal morphology. CLINICAL RELEVANCE: The existence of a radicular groove is closely related to root anatomy and root canal morphology. Anatomical complexity increases the difficulty of root canal treatment and periodontal therapy; therefore, the current data may provide clinicians with a more thorough understanding of the relationship between radicular grooves and root canal morphology.
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Cavidad Pulpar/anatomía & histología , Mandíbula/anatomía & histología , Microtomografía por Rayos X/métodos , HumanosRESUMEN
PURPOSE: The authors conducted a meta-analysis to determine the effect of a history of periodontitis on the long-term survival of dental implants. MATERIALS AND METHODS: An electronic search of PubMed and a supplemental manual search were conducted. Studies published in English through March 2013 were included in the meta-analysis. Survival rates, success rates, periodontal status, types of periodontitis, most recent follow-up time, and other information were extracted and analyzed. RESULTS: Thirteen studies involving 2,011 patients and 6,802 implants were included in the meta-analysis. The results revealed that a history of periodontitis, especially aggressive periodontitis, is associated with significantly higher risks of long-term implant failure versus a healthy periodontium (risk ratio [RR] = 1.03, 95% confidence interval [CI] = 1.02 to 1.04). Based on the limited number of included articles, a subgroup analysis showed that a history of periodontitis had no statistically significant effect on implant survival up to 100 months of follow-up (RR = 1.03, 95% CI = 0.99 to 1.06); however, it did significantly affect implant survival within a period of 101 to 200 months (RR = 1.03, 95% CI = 1.02 to 1.04). Some implant systems also significantly influenced the correlation between a history of periodontitis and implant survival. CONCLUSION: Within the limitations of this meta-analysis, a history of periodontitis is estimated to be a statistical risk factor for the long-term survival of dental implants. This negative effect would be most evident in patients with aggressive periodontitis, severe periodontitis, or after a longer follow-up.
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Implantes Dentales , Fracaso de la Restauración Dental , Periodontitis/complicaciones , Periodontitis Agresiva/complicaciones , Implantes Dentales/efectos adversos , Estudios de Seguimiento , Humanos , Índice Periodontal , Periodontitis/clasificación , Factores de Riesgo , Análisis de SupervivenciaRESUMEN
The formation and attachment of new cementum is crucial for periodontium regeneration. Tissue engineering is currently explored to achieve complete, reliable and reproducible regeneration of the periodontium. The capacity of multipotency and self-renewal makes adipose tissue-deprived stem cells (ADSCs) an excellent cell source for tissue regeneration and repair. After rat ADSCs were cultured in dental follicle cell-conditioned medium (DFC-CM) supplemented with DKK-1, an inhibitor of the Wnt pathway, followed by 7 days of induction, they exhibited several phenotypic characteristics of cementoblast lineages, as indicated by upregulated expression levels of CAP, ALP, BSP and OPN mRNA, and accelerated expression of BSP and CAP proteins. The Wnt/ß-catenin signaling pathway controls differentiation of stem cells by regulating the expression of target genes. Cementoblasts share phenotypical features with osteoblasts. In this study, we demonstrated that culturing ADSCs in DFC-CM supplemented with DKK-1 results in inhibition of ß-catenin nuclear translocation and down-regulates TCF-4 and LEF-1 mRNA expression levels. We also found that DKK-1 could promote cementogenic differentiation of ADSCs, which was evident by the up-regulation of CAP, ALP, BSP and OPN gene expressions. On the other hand, culturing ADSCs in DFC-CM supplemented with 100 ng/mL Wnt3a, which activates the Wnt/ß-catenin pathway, abrogated this effect. Taken together, our study indicates that the Wnt/ß-catenin signaling pathway plays an important role in regulating cementogenic differentiation of ADSCs cultured in DFC-CM. These results raise the possibility of using ADSCs for periodontal regeneration by modifying the Wnt/ß-catenin pathway.
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Tejido Adiposo/citología , Medios de Cultivo Condicionados/farmacología , Saco Dental/citología , Células Madre/citología , Animales , Western Blotting , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Cementogénesis/efectos de los fármacos , Cementogénesis/fisiología , Saco Dental/metabolismo , Inmunohistoquímica , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Células Madre/efectos de los fármacos , Vía de Señalización Wnt/efectos de los fármacosRESUMEN
Despite efforts in peripheral nerve injury and regeneration, it is difficult to achieve a functional recovery following extended peripheral nerve lesions. Even if artificial nerve conduit, cell components and growth factors can enhance nerve regeneration, integration in peripheral nerve repair and regeneration remains yet to be explored. For this study, we used chitosan/gelatin nerve graft constructed with collagenous matrices as a vehicle for Schwann cells and transforming growth factor-ß1 to bridge a 10-mm gap of the sciatic nerve and explored the feasibility of improving regeneration and reinnervation in rats. The nerve regeneration was assessed with functional recovery, electrophysiological test, retrograde labeling, and immunohistochemistry analysis during the post-operative period of 16 weeks. The results showed that the internal sides of the conduits were compact enough to prevent the connective tissues from ingrowth. Nerve conduction velocity, average regenerated myelin area, and myelinated axon count were similar to those treated with autograft (p > 0.05) but significantly higher than those bridged with chitosan/gelatin nerve graft alone (p < 0.05). Evidences from retrograde labeling and immunohistochemistry analysis are further provided in support of improving axonal regeneration and remyelination. A designed graft incorporating all of the tissue-engineering strategies for peripheral nerve regeneration may provide great progress in tissue engineering for nerve repair.
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Quitosano/química , Gelatina/química , Proteínas de la Mielina/metabolismo , Regeneración Nerviosa , Células de Schwann/trasplante , Ingeniería de Tejidos/métodos , Factor de Crecimiento Transformador beta1/uso terapéutico , Animales , Axones/fisiología , Proteína GAP-43/metabolismo , Ratas , Ratas Sprague-Dawley , Células de Schwann/citología , Nervio Ciático/lesiones , Nervio Ciático/metabolismo , Nervio Ciático/patología , Trasplante AutólogoRESUMEN
Apert syndrome is a common craniosynostosis caused by gain-of-function missense mutations of fibroblast growth factor receptor 2 (FGFR2). Mice with the FGFR2 S252W mutation can elucidate the mechanism by which the human Apert syndrome phenotypes arise. However, many studies have focused on mutant skull and long bone malformation, only few studies have focused on mandible changes. Bone formation and micro-architecture between 28- and 56-day-old mutant mice and controls were compared to investigate the changes in the mandibular micro-architecture caused by the Fgfr2(S252W/+) mutation to provide a basis for exploring the pathogenesis and therapeutic measures of human Apert syndrome. Fgfr2(S252W/+) mutant mice were established, and their general characteristics, including weight, naso-anal length, and calcium and phosphate content in serum and bone were tested. Calcein labeling, tartrate-resistant acid phosphatase staining and toluidine blue staining were used to detect osteoblast and osteoclast activities. H&E staining and micro-CT detection were used to test micro-architecture changes. The changes in mineral apposition rate and micro-architecture of the Fgfr2(S252W/+) mice were statistically significant; however, the magnitude of the micro-architecture became less with age. The Fgfr2(S252W/+) mutation may retard mandibular bone formation, decreased bone volume, and compromised skeletal architecture by regulating both osteoblastogenesis and osteoclastogenesis.
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Acrocefalosindactilia/genética , Densidad Ósea/genética , Huesos/metabolismo , Calcio/sangre , Mandíbula/patología , Osteogénesis/genética , Fosfatos/sangre , Receptor Tipo 2 de Factor de Crecimiento de Fibroblastos/genética , Animales , Huesos/patología , Modelos Animales de Enfermedad , Humanos , Mandíbula/metabolismo , Ratones , MutaciónRESUMEN
OBJECTIVES: This study aimed to investigate the root canal morphology of mandibular first premolar teeth in a population from southwestern China by micro-computed tomography (micro-CT). MATERIALS AND METHODS: Human mandibular first premolars (115) were selected and prepared for micro-CT analysis with a slice thickness of 30 µm. Details of root canal orifices, canals, accessory canals, apical foramina-apical delta intercanal communication, loops and isthmuses, and mesial invagination were analyzed from reconstructed three-dimensional (3D) images. RESULTS: Canal patterns categorized according to the classification defined by Vertucci (Endod Top 10:3-29, 2005) as types I (65.2%), III (2.6%), V (22.6%), and VII were identified (0.9%). Accessory canals were present in 35.7% of the samples and were predominantly located in the apical third of the root. A single apical foramen was observed in 50.4% of the samples and two or three foramina in 28.7% and 14.8%, respectively. Apical delta was identified in 6.1% of the samples and the prevalence of intercanal communication and loops was 3.5% and 7%, respectively. Mesial invagination of the root was identified in 27.8% of the samples, the majority of which contained multiple canals. CONCLUSIONS: The data obtained in this study revealed complex root morphology with high prevalence of multiple canals, more than half of which exhibited type I canal patterns. CLINICAL RELEVANCE: Micro-CT was used as a noninvasive technique for 3D investigation of root canal morphology in the mandibular first premolars of a population from southwestern China. Furthermore, data obtained revealed complex anatomy of various types.
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Diente Premolar/anatomía & histología , Cavidad Pulpar/anatomía & histología , Cavidad Pulpar/diagnóstico por imagen , Microtomografía por Rayos X , Diente Premolar/diagnóstico por imagen , China , Femenino , Humanos , Masculino , MandíbulaRESUMEN
Several populations of stem cells, including those from the dental pulp and periodontal ligament, have been isolated from different parts of the tooth and periodontium. The characteristics of such stem cells have been reported as well. However, as a common progenitor of these cells, ectomesenchymal stem cells (EMSCs), derived from the cranial neural crest have yet to be fully characterized. The aim of this study was to better understand the characteristics of EMSCs isolated from rat embryonic facial processes. Immunohistochemical staining showed that EMSCs had migrated to rat facial processes at E11.5, while the absence of epithelial invagination or tooth-like epithelium suggested that any epithelial-mesenchymal interactions were limited at this stage. The p75 neurotrophin receptor (p75NTR), a typical neural crest marker, was used to select p75NTR-positive EMSCs (p75(+) EMSCs), which were found to show a homogeneous fibroblast-like morphology and little change in the growth curve, proliferation capacity, and cell phenotype during cell passage. They also displayed the capacity to differentiate into diverse cell types under chemically defined conditions in vitro. p75(+) EMSCs proved to be homogeneous, stable in vitro and potentially capable of multiple lineages, suggesting their potential for application in dental or orofacial tissue engineering.
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Células Madre Embrionarias/fisiología , Células Madre Mesenquimatosas/fisiología , Células Madre Multipotentes/fisiología , Cresta Neural/citología , Receptores de Factor de Crecimiento Nervioso/análisis , Animales , Biomarcadores/análisis , Diferenciación Celular , Movimiento Celular , Separación Celular , Células Cultivadas , Embrión de Mamíferos/citología , Células Madre Embrionarias/citología , Células Madre Mesenquimatosas/citología , Células Madre Multipotentes/citología , Proteínas del Tejido Nervioso , Ratas , Ratas Sprague-Dawley , Receptores de Factores de Crecimiento , Ingeniería de TejidosRESUMEN
OBJECTIVE: To observe the changes of surface morphology and temperature of dental pulp cavity in vitro after irradiated by Er:YAG laser with different energy and irradiation time. METHODS: All of the 96 samples from 24 teeth in vitro were collected from dental clinical departments then divided into two groups (group A and group B) randomly. We chose the energy of 20 Hz, and 1, 2, 3, 4, 5, 6 W to treat the samples in group A and group B and the irradiation time was 10s or 20s. We recorded the temperature changes of dental pulp cavity by digital thermometer and observe the morphology of tooth enamel by scanning electron microscope (SEM). RESULTS: With the extension of irradiation time and increasing of energy, the temperatures of dental pulp cavity were significantly increased after the treatment of Er: YAG laser. The two groups of tooth enamel surface morphology were changed after irradiated by Er: YAG laser with different energy and irradiation time. However, there was no melting and carbonation on the surface of tooth enamel after the treatment of Er:YAG laser in two groups. CONCLUSION: The temperatures of dental pulp cavity were increased after irradiated by increasing laser energy density fom 1 W to 6 W. No melting or carbonized phenomenon was found in enamel within the energy of 1 W to 6 W. All the data would provide evidences for clinical treatment of cavity.
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Cavidad Pulpar , Láseres de Estado Sólido , Caries Dental , Esmalte Dental , Humanos , Técnicas In Vitro , Rayos Láser , Microscopía Electrónica de Rastreo , TemperaturaRESUMEN
OBJECTIVE: To compare the consistency of root canal configuration types of mandibular first premolar by using micro-CT and radio visio graphy (RVG). METHODS: One hundred extracted mandibular first premolars with complete dental root and apex which received no endodontic treatment were randomly selected. Each tooth was radiographed with RVG through a buccolingual and mesiodistal direction, and then scanned with micro-CT and reconstructed. The classifications of the root canal types according to Vertucci's type with the two methods were compared. RESULTS: The canal patterns were classified as type I (67%), type III (3%), type V (18%), type VII (2%), additional type (10%) with micro-CT and canal patterns as type I (71%), type III (2%), type V (23%), type VII (1%), additional type (3%) with RVG. 63% of teeth showed one canal in both micro-CT and RVG. Only 25% of teeth were diagnosed as complex canal by the same canal type in both micro-CT and RVG. The Kappa value between micro-CT and RVG was 0.541 which suggested that the two kinds of methods had intermediate consistency. 82.8% of the premolars with root groove had two or more than two canals. CONCLUSION: Although RVG can basically reflect the root canal system type of the mandibular first premolars in vitro, it offers poor accuracy images to complex root canals. Micro-CT three-dimensional images could clearly and precisely display the root canal system morphology of the mandibular first pre-molars in vitro.
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Diente Premolar , Microtomografía por Rayos X , Cavidad Pulpar , Humanos , Mandíbula , Diente Molar , Tratamiento del Conducto Radicular , Raíz del DienteRESUMEN
INTRODUCTION: Little is known about the lingual canal in the Vertucci type V mandibular first premolar. This study investigated the location of the lingual canal orifice and the curvature of the lingual canal by using micro-computed tomography. METHODS: One hundred fifteen mandibular first premolars were scanned by micro-computed tomography, reconstructed 3-dimensionally by using Mimics 10.01 software, and displayed in parallel projection mode. Twenty-six teeth with Vertucci type V canal were selected for further study. The lingual canal orifice was located by measurements made in both lingual and proximal views. The angle alpha (α) between the start of the lingual canal and the main canal and the angle beta (ß) of the curvature of lingual canal were also measured. RESULTS: In proximal view, 69% of lingual canals were located in the middle third of the tooth and the remainder in the apical third. In lingual view, 73% were located in the middle third of the root and the remainder in the coronal third. Mean angle α and angle ß were 33.54° and 26.66°, respectively, in proximal view and 8.31° and 11.31°, respectively, in lingual view (P < .05). The highest values of angles α and ß were observed in proximal view (65.24°and 43.39°, respectively). In most cases, angles α and ß were severely curved in proximal view and straight or only slightly curved in lingual view. CONCLUSIONS: Detailed information on the lingual canal is essential for successful endodontic treatment in patients with mandibular first premolar. The view used for imaging influences the information obtained.
Asunto(s)
Diente Premolar/diagnóstico por imagen , Cavidad Pulpar/diagnóstico por imagen , Mandíbula/diagnóstico por imagen , Microtomografía por Rayos X/métodos , Diente Premolar/anomalías , Cavidad Pulpar/anomalías , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Imagenología Tridimensional/métodos , Odontometría , Ápice del Diente/diagnóstico por imagen , Microtomografía por Rayos X/instrumentaciónRESUMEN
Periodontal ligament stem cells (PDLSCs), a new population of mesenchymal stem cells (MSCs), have been isolated from the periodontal ligament (PDL). The capacity of multipotency and self-renewal makes them an excellent cell source for bone regeneration and repair. However, their bone-regeneration ability could be awakened in inflammatory microenvironments, which may be the result of changes in their differentiation potential. Recently, genetic evidences has shown that the Wnt pathway plays an important role in bone homeostasis. In this study we have determined the specific role of ß-catenin in osteogenic differentiation of PDLSCs obtained from inflammatory microenvironments (P-PDLSCs). The inflammatory microenvironment, while inhibiting osteogenic differentiation potential, promotes proliferation of MSCs. A higher the level of ß-catenin in P-PDLSCs than in H-PDLSCs (PDLSCs obtained from a healthy microenvironment) resulted in the same disparity in canonical Wnt signaling pathway activation between each cell type. Here we show that activation of ß-catenin suppresses the noncanonical Wnt/Ca(2+) pathway, leading to increased proliferation but reduced osteogenic differentiation of P-PDLSCs. Downregulation of the levels of ß-catenin by treatment with dickkopf-1 (DKK-1) leads to activation of the noncanonical Wnt/Ca(2+) pathway, which, in turn, results in the promotion of osteogenic differentiation in P-PDLSCs. Interestingly, ß-catenin can affect both the canonical Wnt/ß-catenin pathway and the noncanonical Wnt/Ca(2+) pathway. Our data indicate that ß-catenin plays a central role in regulating osteogenic differentiation of MSCs in inflammatory microenvironments. Given the important role of Wnt signaling in osteogenic differentiation, it is possible that agents that can modify this pathway may be of value in bone regeneration by MSCs in chronic inflammatory microenvironments.
Asunto(s)
Diferenciación Celular , Microambiente Celular , Inflamación/patología , Osteogénesis , Células Madre/patología , Vía de Señalización Wnt , beta Catenina/metabolismo , Adulto , Calcio/metabolismo , Ciclo Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Separación Celular , Microambiente Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Citometría de Flujo , Humanos , Péptidos y Proteínas de Señalización Intercelular/farmacología , Osteogénesis/efectos de los fármacos , Ligamento Periodontal/patología , Células Madre/efectos de los fármacos , Células Madre/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Vía de Señalización Wnt/efectos de los fármacos , Proteína Wnt3A/farmacologíaRESUMEN
Adipose tissue-derived stem cells (ADSCs), which are easily harvested and show excellent pluripotency potential, have generated considerable interest in regenerative medicine. In this study, the differentiation of ADSCs was assessed after treatment with dental follicle cell conditioned medium (DFCCM) containing dentin non-collagenous proteins (dNCPs). ADSCs exhibited a fibroblast-like morphology and high proliferative capacity. However, after treatment with dNCPs/DFCCM, ADSCs changed from a fibroblast-like to cementoblast-like morphology and significantly lost their proliferative capacity. Alkaline phosphatase activity and in vitro mineralization behaviour of ADSCs were significantly enhanced. Mineralization-related markers including cementum attachment protein, bone sialoprotein, osteocalcin, osteopontin and osteonectin were detected at mRNA or protein levels, whereas dentin sialophosphoprotein and dentin sialoprotein were not detected, implying a cementoblast-like phenotype. These results demonstrate that ADSCs acquired cementoblast features in vitro with dNCPs/DFCCM treatment and could be a potential source of cementogenic cells for periodontal regeneration.