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Despite the understanding of the coronavirus disease-19 (COVID-19), the role of salivary extracellular vesicles (sEVs) in COVID-19 remains unclear. Exploring the proteomic cargo of sEVs could prove valuable for diagnostic and prognostic purposes in assessing COVID-19. The proteomic cargo of sEVs from COVID-19(+) subjects and their healthy close contacts (HCC) was explored. sEVs were isolated by ultracentrifugation from unstimulated saliva samples, and subsequently characterized through nanoparticle tracking, transmission electron microscopy, and Western blot analyses. The proteomic cargo of sEVs was processed by LC-MS/MS. sEVs were morphologically compatible with EVs, with the presence of Syntenin-1 and CD81 EV markers. The sEV pellet showed 1417 proteins: 1288 in COVID-19(+) cases and 1382 in HCC. In total, 124 proteins were differentially expressed in sEVs from COVID-19(+) subjects. "Coronavirus-disease response", "complement and coagulation cascades", and "PMN extracellular trap formation" were the most enriched KEGG pathways in COVID-19(+) cases. The most represented biological processes were "Hemoglobin and haptoglobin binding" and "oxygen carrier activity", and the best-denoted molecular functions were "regulated exocytosis and secretion" and "leucocyte and PMN mediated immunity". sEV proteomic cargo in COVID-19(+) suggests activity related to immune response processes, oxygen transport, and antioxidant mechanisms. In contrast, in HCC, sEV signature profiles are mainly associated with epithelial homeostasis.
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COVID-19 , Vesículas Extracelulares , Humanos , Cromatografía Liquida , Proteómica , Espectrometría de Masas en Tándem , OxígenoRESUMEN
INTRODUCTION: Modern tissue engineering strategies have elucidated the potential of regenerative endodontic treatment (RET) as an alternative for treating mature teeth. METHODS: Here, we report two cases in which cell-based RET (CB-RET) using encapsulated allogeneic umbilical cord mesenchymal stem cells (UC-MSCs) in a platelet-poor plasma (PPP)-based scaffold was used in two mature teeth with pulp necrosis and apical periodontitis. RESULTS: After 5 years of follow-up, the healing response was satisfactory in both cases, with evidence of pulp revitalization. CONCLUSIONS: This is the first study to report the success of an extended, 5-year follow-up for allogeneic CB-RET. This report presents an innovative and sustainable solution to challenging endodontic scenarios.
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Necrosis de la Pulpa Dental , Periodontitis Periapical , Endodoncia Regenerativa , Humanos , Necrosis de la Pulpa Dental/terapia , Trasplante de Células Madre Mesenquimatosas/métodos , Periodontitis Periapical/terapia , Endodoncia Regenerativa/métodos , Tratamiento del Conducto Radicular/métodos , Ingeniería de Tejidos/métodos , Andamios del Tejido , Ápice del DienteRESUMEN
The aim of this report is to present the results of a personalised endodontic treatment for an immature molar, using regenerative endodontic procedure (REP) combined with non-surgical root canal treatment (NSRCT), with 7 years of follow-up. The tooth#3 presented combined endodontic pathologies in each root, with different pulpal and periapical status. A REP in the palatal (P) canal and a NSCRT in the mesio-buccal (MB) and disto-buccal (DB) canals were performed. Absence of clinical signs/symptoms and continuous palatal root development with apical closure were observed over 4-years. After 5-years an apical lesion in the MB root was observed. Both MB canals were selectively retreated by completely filling them with TotalFill BC RRM Fast Set putty. After 7-years, the tooth remained functional and the apical lesion was resolving. The palatal root was completely mature. This case report reveals the potential for use of combined treatment approaches for immature multirooted teeth.
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Cavidad Pulpar , Endodoncia Regenerativa , Humanos , Cavidad Pulpar/diagnóstico por imagen , Endodoncia Regenerativa/métodos , Estudios de Seguimiento , Necrosis de la Pulpa Dental/terapia , Tratamiento del Conducto Radicular/métodos , Diente Molar/cirugíaRESUMEN
Regenerative endodontic procedures rely on the delivery of mesenchymal stem cells into the root canal and on the effect of local growth factors from the dentin and blood clot. The aim of this study was to assess the effect of dentin conditioning with ethylenediamine tetraacetic acid (EDTA) and diode lasers with different wavelengths (808 nm and 980 nm) on the expression of odontoblast-like cell markers. Forty dentin cylinders were divided into four groups according to the irrigation protocol: EDTA, EDTA + 808 nm diode laser, EDTA + 980 nm diode laser, and phosphate-buffered saline as the control group. Dental pulp stem cells were seeded into the previously conditioned cylinders and incubated for 14 days. The quantitative real-time polymerase chain reaction was used to evaluate the expression of dentin sialophosphoprotein (DSPP), dentin morphoprotein-1 (DMP-1), and transforming growth factor-beta 1 (TGF-ß1). Data analysis was performed using the Kruskal-Wallis test. The activation of EDTA with 980 nm and 808 nm diode lasers resulted in lower DSPP and DMP-1 expression than that for EDTA alone (p < 0.05 and p < 0.01, respectively). The expression of TGF was similar among all groups. The highest level of expression of odontoblast-like differentiation markers was observed with EDTA alone. However, the use of an 808 nm diode laser during EDTA irrigation reduced the expression of odontoblastic differentiation markers.
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The main goal of regenerative endodontics procedures (REPs) is to revitalize teeth by the regeneration of healthy dental pulp. In this study, we evaluated the potential of combining a natural and accessible biomaterial based on Platelet Poor Plasma (PPP) as a support for dental pulp stem cells (DPSC) and umbilical cord mesenchymal stem cells (UC-MSC). A comparison study between the two cell sources revealed compatibility with the PPP based scaffold with differences noted in the proliferation and angiogenic properties in vitro. Additionally, the release of growth factors including VEGF, HGF and DMP-1, was detected in the media of cultured PPP and was enhanced by the presence of the encapsulated MSCs. Dentin-Discs from human molars were filled with PPP alone or with MSCs and implanted subcutaneously for 4 weeks in mice. Histological analysis of the MSC-PPP implants revealed a newly formed dentin-like structure evidenced by the expression of Dentin sialophosphoprotein (DSPP). Finally, DPSC induced more vessel formation around the dental discs. This study provides evidence of a cost-effective, xenofree scaffold that is compatible with either autologous or allogenic strategy for dental pulp regeneration. This attempt if successfully implemented, could make REPs treatment widely accessible, contributing in improving global health conditions.
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Pulpa Dental/fisiología , Regeneración , Andamios del Tejido , Animales , Pulpa Dental/citología , Femenino , Humanos , Recién Nacido , Masculino , Células Madre Mesenquimatosas/fisiología , Ratones , Microscopía Electrónica de Rastreo , Neovascularización Fisiológica , Plasma , Cordón Umbilical/citología , Adulto JovenRESUMEN
This report describes a regenerative endodontic procedure of an immature permanent incisor with internal root resorption (IRR) and 4-years follow-up. A healthy 8-year-old man was referred for treatment of tooth #9 after a traumatic intrusion. The periapical radiograph showed an IRR and an open apex with periradicular lesion. A diagnosis of pulp necrosis and chronic apical abscess was achieved. In the first appointment, under local anesthesia and rubber dam isolation, an access cavity was designed and the root canal was chemically cleaned under irrigation with 10 mL 1.5% sodium hypochlorite (NaOCl). The root canal was then dried and calcium hydroxide paste was placed. During the second appointment, the root canal was irrigated with 5 mL of 17% ethylenediaminetetraacetic acid (EDTA) for 5 minutes and dried. The blood clot was established in a time of 3 minutes after the bleeding from the periapical tissue was trigged. White mineral trioxide aggregate (MTA) was placed up to the amelocemental junction and the final restoration of the access cavity was carried out. During periodic clinical and radiographic follow-up, the patient remained symptom free, the periapical region was completely healed, inhibition of the root resorption process achieved, and formation of the new periodontal ligament as well as tooth widening development observed, meeting functional expectations after 48 months. The regenerative endodontic procedures are an available option to treat IRR in severely immature teeth. The available literature on the regenerative endodontic procedures applied to IRR treatment is limited, and more research is needed in this field.
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INTRODUCTION: Dental pulp stem cells (DPSC) are very attractive in regenerative medicine. In this study, we focused on the characterization of the functional properties of mesenchymal stem cells derived from DPSCs. Currently, it is unknown whether inflammatory conditions present in an inflamed dental pulp tissue could alter the immunomodulatory properties of DPSCs. This study aimed to evaluate the immunomodulatory capacity in vitro of DPSCs derived from healthy and inflamed dental pulp. METHODS: DPSCs from 10 healthy and inflamed dental pulps (irreversible pulpitis) were characterized according to the minimal criteria of the International Society for Cell Therapy, proliferation, differential potential, and colony-forming units. Furthermore, the immunomodulatory capacity of DPSCs was tested on the proliferation of T lymphocytes by flow cytometry and the in vitro enzyme activity of indoleamine 2, 3-dioxygenase. RESULTS: There were no significant differences in the DPSC characteristics and properties such as immunophenotype, tridifferentiation, colony-forming units, and proliferation of the DPSCs derived from normal and inflamed pulp tissue. Furthermore, there were significant differences in the immunomodulatory capacity of DPSCs obtained from human healthy dental pulp and with the diagnosis of irreversible pulpitis. CONCLUSIONS: Our results showed that DPSCs isolated from inflamed dental pulp showed typical characteristics of MSCs and diminished immunosuppressive capacity in vitro in comparison with MSCs derived from healthy dental pulp. Further investigation in vivo is needed to clarify the mechanism of this diminished immunosuppressive capacity.
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Células Madre Mesenquimatosas , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Pulpa Dental , HumanosRESUMEN
INTRODUCTION: Regenerative endodontic procedures have emerged as a new treatment. The aim of this case report was to describe a regenerative autologous cellular therapy using mesenchymal stem cells from inflamed dental pulp and leukocyte platelet-rich fibrin (L-PRF) in a mature tooth. METHODS: A healthy 50-year-old man consulting for spontaneous dental pain was referred for endodontic treatment in tooth #28, which was diagnosed with symptomatic irreversible pulpitis. Inflamed dental pulp was extracted and transported to a good manufacturing practice laboratory for the isolation and culture of dental pulp stem cells (DPSCs). L-PRF was obtained from the patient's blood and was introduced into the instrumented and disinfected root canal, and expanded DPSCs were inoculated into the clot. The cervical part of the root canal was sealed with Biodentine (Septodont, Saint-Maur-des-Fosses, France) and a composite resin. RESULTS: Follow-up examinations were performed 6 months and 3 years later. The examinations included periapical radiographs (to measure the periapical index [PAI]), cone-beam computed tomographic (CBCT) imaging, sensitivity, and vitality tests. Clinical evaluations revealed normal responses to percussion and palpation tests. The tooth had a delayed response to cold, and the electric pulp test was responsive. The PAI and CBCT imaging revealed that the periapical area remained normal with a PAI score of 1 and a CBCT PAI score of 0. The vitality test performed indicated low blood perfusion units. CONCLUSIONS: This case study reveals the potential use of a patient's own DPSCs and L-PRF as an alternative procedure for the treatment of pulpitis in mature permanent teeth. It also paves the way for the design of personalized cell-based clinical trials in regenerative endodontics.
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Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Pulpa Dental/citología , Leucocitos , Fibrina Rica en Plaquetas , Medicina de Precisión/métodos , Pulpitis/terapia , Endodoncia Regenerativa/métodos , Trasplante de Células Madre/métodos , Células Madre , Células Cultivadas , Humanos , Masculino , Persona de Mediana Edad , Ingeniería de Tejidos/métodos , Trasplante Autólogo , Resultado del TratamientoRESUMEN
High donor variation makes comparison studies between different dental sources dubious. Dental tissues offer a rare opportunity for comparing the biological characteristics of haploidentical mesenchymal stem cells (MSCs) isolated from the same donor. The objective was to identify the optimal dental source of MSCs through a biological and functional comparison of haploidentical MSCs from gingival (GMSCs) and dental pulp stem cells (DPSCs) focusing mainly on their angiogenic potential. The comparison study included (1) surface markers expression, (2) mesodermal differentiation capacity (chondrogenic, adipogenic, and osteogenic), (3) proliferation, (4) migration potential, (5) ability to form colony units, and (6) angiogenic potential in vitro and in vivo. Comparative analysis showed no difference in the immunophenotypic profile nor for the trilineage differentiation potential. Proliferation of GMSCs was higher than DPSCs at day 6 (2.6-fold higher, P < 0.05). GMSCs showed superior migratory capacity compared to DPSCs at 4, 8, and 12 h (2.1-, 1.5-, and 1.2-fold higher, respectively, P < 0.05). Furthermore, GMSCs formed a higher number of colony units for both cell concentrations (1.7- and 1.4-fold higher for 150 and 250 starting cells, respectively, P < 0.05). GMSCs showed an improved angiogenic capacity compared to DPSCs (total tube lengths 1.17-fold higher and 1.5-fold total loops, P < 0.05). This was correlated with an enhanced release of vascular growth factor under hypoxic conditions. Finally, in the plug transplantation assay evaluating the angiogenesis in vivo, the DPSC and GMSC hemoglobin content was 3.9- and 4-fold higher, respectively, when compared to the control (Matrigel alone). GMSCs were superior to their haploidentical DPSCs in proliferation, migration, and angiogenic potentials. This study positions GMSCs in the forefront of dental cell sources for applications in regenerative medicine.
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Movimiento Celular , Proliferación Celular , Pulpa Dental/citología , Encía/citología , Células Madre Mesenquimatosas/citología , Neovascularización Fisiológica , Adolescente , Adulto , Separación Celular , Células Cultivadas , Femenino , Humanos , Masculino , Adulto JovenRESUMEN
OBJECTIVE: To describe mandibular premolar root and canal morphology and its variability in Chilean and Belgian samples using cone-beam computed tomography. METHODS: A cross-sectional and descriptive study was conducted. A total of 402 mandibular premolars were examined using cone-beam computed tomography images of Chilean and Belgian patients. Premolars that met the inclusion criteria were studied in relation to the number of roots, number of canals, root canal configuration, presence of C-shaped configuration, tooth length, and root length. RESULTS: The mandibular first premolar frequently presented with one root (94% Chilean; 100% Belgian), as did the mandibular second premolar (99% Chilean; 98% Belgian). One canal was present in 69% of Chilean, and 83% of Belgian mandibular first premolars, and in 95% of Chilean and 91% of Belgian second premolars. A type I root canal configuration was found in 69% of Chilean and 83% of Belgian first premolars and in 95% of Chilean and 92% of Belgian second premolars. CONCLUSIONS: The anatomical parameters analyzed in the Chilean and Belgian samples are similar. However, it should be noted that there are anatomical variations in mandibular premolars.
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Diente Premolar/diagnóstico por imagen , Tomografía Computarizada de Haz Cónico/métodos , Raíz del Diente/diagnóstico por imagen , Adulto , Bélgica , Chile , Estudios Transversales , Femenino , Humanos , Masculino , Interpretación de Imagen Radiográfica Asistida por ComputadorRESUMEN
Regenerative endodontic procedures (REPs) associated with apical surgery could represent an alternative treatment strategy for patients whose teeth present incomplete root formation and extensive apical lesions. Leukocyte platelet-rich fibrin (L-PRF) has potential benefits in REPs; it could promote apical root formation and optimal bone healing. The aim of this case report was to describe innovative regenerative endodontic therapy using L-PRF in the root canal and an extensive apical lesion in an immature tooth with dens invaginatus and asymptomatic apical periodontitis. A healthy 20-year-old woman was referred to the dental clinic of the Universidad de Los Andes, Santiago, Chile, for endodontic treatment in tooth # 22 with incomplete root development and an extensive apical lesion. The diagnosis was asymptomatic apical periodontitis associated with dens invaginatus type II. The patient was treated with an innovative approach using L-PRF in REPs associated with apical surgery. Follow-ups were performed at 6 months and 1 year later. They included periapical radiographs, cone-beam computed tomographic imaging, sensitivity, and vitality tests. The clinical evaluations performed at 6 months and 1 year revealed an absence of symptoms. The radiographic evaluations showed that the apical lesion was resolved. The cone-beam images indicated that the root length increased and the walls had thickened. The sensitivity tests were positive, and the laser Doppler flowmetry showed positive blood flow after 1 year. The success of the results in this case report indicate that L-PRF can be used as a complement in apical surgery and REPs and could provide an innovative alternative treatment strategy for complex clinical cases like these.
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Regeneración Tisular Dirigida/métodos , Leucocitos/fisiología , Fibrina Rica en Plaquetas/fisiología , Ápice del Diente/fisiología , Terapia Combinada , Tomografía Computarizada de Haz Cónico , Femenino , Humanos , Periodontitis Periapical/diagnóstico por imagen , Periodontitis Periapical/cirugía , Radiografía Dental , Regeneración , Ápice del Diente/diagnóstico por imagen , Ápice del Diente/cirugía , Adulto JovenRESUMEN
BACKGROUND: The correct estimation of the post mortem interval (PMI) can be crucial on the success of a forensic investigation. Diverse methods have been used to estimate PMI, considering physical changes that occur after death, such as mortis algor, livor mortis, among others. Degradation after death of dental pulp is a complex process that has not yet been studied thoroughly. It has been described that pulp RNA degradation could be an indicator of PMI, however this study is limited to 6 days. The tooth is the hardest organ of the human body, and within is confined dental pulp. The pulp morphology is defined as a lax conjunctive tissue with great sensory innervation, abundant microcirculation and great presence of groups of cell types. AIM: The aim of this study is to describe the potential use of pulp post mortem alterations to estimate PMI, using a new methodology that will allow obtainment of pulp tissue to be used for histomorphological analysis. The current study will identify potential histological indicators in dental pulp tissue to estimate PMI in time intervals of 24h, 1 month, 3 months and 6 months. MATERIALS AND METHOD: This study used 26 teeth from individuals with known PMI of 24h, 1 month, 3 months or 6 months. All samples were manipulated with the new methodology (Carrasco, P. and Inostroza C. inventors; Universidad de los Andes, assignee. Forensic identification, post mortem interval estimation and cause of death determination by recovery of dental tissue. United State patent US 61/826,558 23.05.2013) to extract pulp tissue without the destruction of the tooth. The dental pulp tissues obtained were fixed in formalin for the subsequent generation of histological sections, stained with Hematoxylin Eosin and Masson's Trichrome. All sections were observed under an optical microscope using magnifications of 10× and 40×. RESULTS: The microscopic analysis of the samples showed a progressive transformation of the cellular components and fibers of dental pulp along PMI. These results allowed creating a chart of qualitative and quantitative parameters to be used on the estimation on PMI based on microscopic degradation of dental pulp. CONCLUSIONS: The histological transformations of dental pulp as a function of time can be used as PMI indicators.
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Pulpa Dental/patología , Cambios Post Mortem , Adulto , Femenino , Patologia Forense , Humanos , Masculino , Microscopía , Proyectos Piloto , Adulto JovenRESUMEN
INTRODUCTION: Direct pulp capping treatment is intended to preserve pulp vitality, to avoid or retard root canal treatment, and, in cases with an open apex, to allow continued root development. Historically, calcium hydroxide (CH) was the gold standard material, but nowadays calcium silicate materials (CSMs) are displacing CH because of their high bioactivity, biocompatibility, sealing ability, and mechanical properties. However, more randomized clinical trials are needed to confirm the appropriateness of CSMs as replacement materials for CH in direct pulp capping procedures. METHODS: A randomized clinical trial was conducted that included 169 patients (mean age, 11.3 years) from the Maipo district (Chile). The inclusion criterion was patients with 1 carious permanent tooth with pulpal exposure, a candidate for a direct pulp capping procedure. The patients were randomly allocated to one of the experimental groups (CH, Biodentine, or mineral trioxide aggregate [MTA]). Clinical follow-up examinations were performed at 1 week, 3 months, 6 months, and 1 year. The Fisher exact test was performed. RESULTS: At the follow-up examination at 1 week, the patients showed 100% clinical success. At 3 months, there was 1 failure in the CH group. At 6 months, there were 4 new failures (1 in the CH group and 3 in the MTA group). At 1 year, there was another failure in the CH group. There were no statistically significant differences among the experimental groups. CONCLUSIONS: CSMs appear to be suitable materials to replace CH. Although no significant differences were found among the materials studied, Biodentine and MTA offered some advantages over CH.
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Compuestos de Aluminio/uso terapéutico , Compuestos de Calcio/uso terapéutico , Hidróxido de Calcio/uso terapéutico , Caries Dental/cirugía , Recubrimiento de la Pulpa Dental/métodos , Óxidos/uso terapéutico , Materiales de Recubrimiento Pulpar y Pulpectomía/uso terapéutico , Silicatos/uso terapéutico , Niño , Dentición Permanente , Combinación de Medicamentos , Femenino , Humanos , Masculino , Diente Molar/cirugíaRESUMEN
Objectives: To implement a dentin slice model of mesenchymal stem cells derived from dental tissues in a fibrin-agarose construct for dental pulp regeneration. Material and Methods: MSCs derived from different oral cavity tissues were combined with a fibrin-agarose construct at standard culture conditions. Cell viability and proliferation tests were assayed using a fluorescent cell dye Calcein/Am and WST-1 kit. The proliferation assay was evaluated at 24, 48, 72, and 96 hours. Also, we assessed the dental pulp stem cells (DPSCs) cell morphology inside the construct with histological stains such as Hematoxylin and Eosin, Masson's trichrome, and Periodic acidSchiff. In addition, we elaborated a tooth dentin slice model using a culture of DPSC in the fibrinagarose constructs co-adhered to dentin walls. Results: The fibrin-agarose construct was a biocompatible material for MSCs derived from dental tissues. It provided good conditions for MSCs' viability and proliferation. DPSCs proliferated better than the other MSCs, but the data did not show significant differences. The morphology of DPSCs inside the construct was like free cells. The dentin slice model was suitable for DPSCs in the fibrin-agarose construct. Conclusion: Our findings support the dentin slice model for future biological use of fibrin-agarose matrix in combination with DPSCs and their potential use in dental regeneration. The multipotency, high proliferation rates, and easy obtaining of the DPSCs make them an attractive source of MSCs for tissue regeneration.
Objetivos: Implementar un modelo de dentina con células madre mesenquimales derivadas de tejidos dentales en una constructo de fibrina-agarosa para la regeneración de la pulpa dental. Material y Métodos: Las MSC derivadas de diferentes tejidos de la cavidad oral se combinaron con una construcción de fibrina-agarosa en condiciones de cultivo estándar. Las pruebas de viabilidad y proliferación celular se ensayaron utilizando un kit de colorante celular fluorescente Calcein/Am y WST-1. El ensayo de proliferación se evaluó a las 24, 48, 72 y 96 horas. Además, evaluamos la morfología celular de las células madre de la pulpa dental (DPSC) dentro de la construcción con tinciones histológicas como hematoxilina y eosina, tricrómico de Masson y ácido peryódico de Schiff. Además, elaboramos un modelo de rebanadas de dentina dental utilizando un cultivo de DPSC en las construcciones de fibrina-agarosa coadheridas a las paredes de la dentina. Resultados: La construcción de fibrina-agarosa fue un material biocompatible para las MSC derivadas de tejidos dentales. Proporcionó buenas condiciones para la viabilidad y proliferación de las MSC. Las DPSC proliferaron mejor que las otras MSC, pero los datos no mostraron diferencias significativas. La morfología de las DPSC dentro de la construcción era como la de las células libres. El modelo de corte de dentina fue adecuado para DPSC en la construcción de fibrina-agarosa.Conclusión: Nuestros hallazgos respaldan el modelo de corte de dentina para el futuro uso biológico de la matriz de fibrina-agarosa en combinación con DPSC y su uso potencial en la regeneración dental. El multipotencial, las altas tasas de proliferación y la fácil obtención de las DPSC las convierten en una fuente atractiva de MSC para la regeneración de tejidos.
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Humanos , Sefarosa/química , Células Madre/química , Materiales BiocompatiblesRESUMEN
PURPOSE: This study used cone-beam computed tomography (CBCT) to characterize mandibular molar root and canal morphology and its variability in Belgian and Chilean population samples. MATERIALS AND METHODS: We analyzed the CBCT images of 515 mandibular molars (257 from Belgium and 258 from Chile). Molars meeting the inclusion criteria were analyzed to determine (1) the number of roots; (2) the root canal configuration; (3) the presence of a curved canal in the cross-sectional image of the distal root in the mandibular first molar and (4) the presence of a C-shaped canal in the second mandibular molar. A descriptive analysis was performed. The association between national origin and the presence of a curved or C-shaped canal was evaluated using the chi-squared test. RESULTS: The most common configurations in the mesial root of both molars were type V and type III. In the distal root, type I canal configuration was the most common. Curvature in the cross-sectional image was found in 25% of the distal canals of the mandibular first molars in the Belgian population, compared to 11% in the Chilean population. The prevalence of C-shaped canals was 10% or less in both populations. CONCLUSION: In cases of unclear or complex root and canal morphology in the mandibular molars, CBCT imaging might assist endodontic specialists in making an accurate diagnosis and in treatment planning.
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MatrigelBD is a hydrogel scaffold with three-dimensional intercrossed networks of hydrophilic polymers with high water content. Human gingival tissue might represent a better source of MSCs, allowing these cells to be easily obtained in a relatively non-invasive way. The objective of this study was to evaluate the biocompatibility of MatrigelBD with GMSCs in vitro. Gingival connective tissue samples were obtained from healthy donors. Fresh tissue was minced and cultured during two weeks, after which cells at passage fourth were analyzed for their immune phenotype by flow cytometry. Differentiation into osteogenic, chondrogenic, and adipogenic lineages was induced and evaluated by culture staining. The "construct" was made of MatrigelBD with GMSC. To assess the biocompatibility, an MTT cellular proliferation assay was performed. The differentiation potential of the cells toward the osteogenic, adipogenic, and chondrogenic lineages was analyzed after 21 days of growth in MatrigelBD with induction differentiation media. The MTT analysis showed that MatrigelBD stimulated cell proliferation; the GMSCs maintained the expression of MSC markers. Importantly, the growth of GMSCs within the MatrigelBD did not interfere with the cell differentiation potential. These findings indicate that MatrigelBD is biocompatible with GMSCs, and this matrix improves cell proliferation in vitro.
MatrigelBD es un andamiaje de hidrogel con redes tridimensionales entrecruzadas de polímeros hidrófilos con un alto contenido de agua. El tejido gingival humano podría representar una mejor fuente de MSCs, estas células pueden obtenerse fácilmente de una manera relativamente no invasiva. El objetivo de este estudio fue evaluar la biocompatibilidad de MatrigelBD con GMSCs in vitro. Muestras gingivales de tejido conectivo se obtuvieron de donantes sanos. El tejido se trituró y se cultivó durante dos semanas, y cuando las células se encontraban en el cuarto pasaje se les analizó su fenotipo inmunológico utilizando citometría de flujo. Se indujo la diferenciación hacia los linajes osteogénico, condrogénico y adipogénico, evaluandose con tinciones. El "constructo" se hizo de MatrigelBD con GMSC. Para evaluar la biocompatibilidad, se realizó un ensayo de proliferación celular: MTT. Se analizó el potencial de diferenciación de las células hacia los linajes osteogénico, adipogénico y condrogénico después de 21 días de cultivo en MatrigelBD con medio de diferenciación de inducción. El análisis de MTT mostró que MatrigelBD estimula la proliferación celular; GMSCs mantiene la expresión de marcadores de MSC. Es importante destacar que el crecimiento de GMSCs en MatrigelBD no interfirió con el potencial de diferenciación celular. Estos hallazgos indican que MatrigelBD es biocompatible con GMSCs, y esta matriz mejora la proliferación celular in vitro.