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Purpose: This study aimed to evaluate the cytotoxicity of a biocompatible 3D-printed resin material for occlusal devices after post-processing with two different high-intensity UV-polymerization devices and two rinsing solvents, in the presence of human gingival fibroblasts (HGFs)., Material and Methods: Sample discs from the 3D-printed resin material were printed (2 mm in height and 6 mm in diameter [N = 40]) and divided into 4 groups (n = 10) based on post-processing methods: a high-intensity UV polymerization device with isopropyl alcohol, a high-intensity UV polymerization device with a modified glycol solvent, a UV cleaning and curing unit with isopropyl alcohol, a UV cleaning and curing unit with a modified glycol solvent, and a control group cultured in DMEM medium. Different tests were performed to evaluate their cytocompatibility on HGFs: MTT assay, cell migration assay, cell cytoskeleton staining, scanning electron microscopy (SEM), and cell apoptosis and generation of intracellular reactive oxygen species (ROS). Statistical analyses were performed using one-way ANOVA and the Tukey post hoc test (α = 0.05)., Results: Cytocompatibility, MTT assay, cell migration assay, cell cytoskeleton staining, and SEM images were similar, regardless of the post-processing protocol, compared with the control group., Conclusions: No differences were found in the cytotoxicity of the 3D-printed resin material for occlusal devices after the following post-processing methods: two different UV-polymerization devices and two rinsing solvents (isopropyl alcohol and a modified glycol solvent)., (© 2024 The Author(s). Journal of Prosthodontics published by Wiley Periodicals LLC on behalf of American College of Prosthodontists.)

Introduction: The aim of this study was to compare the biological effects of four chemical caries removal materials and to assess their cytotoxicity using human dental pulp stem cells (hDPSCs)., Methods: The products evaluated are: 1 - papain-based product (BRIX 3000®); 2 - papain/chloramine based products (NATURAL-CARE and Papacárie Duo®); and 3 - chloramine based product (Cariesolut). The following in vitro experiments were carried out: IC50 measurement, cell metabolic activity (MTT) assay, cell migration, immunofluorescence experiment, cell apoptosis analysis, and reactive oxygen species (ROS) production analysis. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p < 0.05)., Results: The IC50 values were: Brix 3000: 0.596%; Papacárie Duo: 0.052%; NATURAL CARE: 1.034%; and Cariesolut: 0.020%. The MTT assays showed non-adequate cell viability of all chemical-mechanical caries removal tested at 2% at 24, 48, and 72 h (p < 0.001). The same behaviour was observed at 0.1% in the Papacárie Duo and Cariesolut groups. In contrast, 0.1% of Brix 3000 at all times and NATURAL CARE at 24 h treated cells showed cell viability rates similar to the control group. At 0.01% only Brix 3000 did not show statistically significant differences at any time. Delayed cell migration was observed in all hDPSCs treated with Papacárie Duo and Cariesolut (p < 0.01 and p < 0.001). Phalloidin staining images showed a high confluence of cells in the presence of NATURAL CARE, similar to the control group. On the contrary, no cells were observed in Brix 3000 and Cariesolut at 2% and 0.1% concentrations. Papacárie Duo showed cells at all concentrations, but hDPSCs treated at 0.01% concentration exhibited better proliferation and spreading than those in the control group. Apoptosis essay showed that Brix 3000 at both 0.1% and 0.01% had a percentage of live cells higher than 99%, with 68.4% live cells at 2%, 3.69% early apoptotic cells, and 27.9% late apoptotic cells. Conversely, the rest of the materials showed an abundance of apoptotic cells, even at low concentrations. 0.1% and 0.01% of BRIX 3000 did not affect the ROS production levels, while 2% of BRIX 3000 counterpart very significantly increased the percentage of CM-H2DCFDA positive cells. Again, all concentrations of Cariesolut showed significantly higher levels of ROS production than those observed in control cells., Conclusion: Our results suggest that Brix 3000 would be the most suitable material for chemical caries removal, with Papacárie Duo and NATURAL CARE also being good options, and discourage the use of Cariesolut due to its low cytocompatibility on dental pulp stem cells., (© 2024 S. Karger AG, Basel.)

To evaluate the effects of premixed calcium silicate based ceramic sealers on the viability and osteogenic/cementogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The materials evaluated were TotalFill BC Sealer (TFbc), AH Plus Bioceramic Sealer (AHPbc), and Neosealer Flo (Neo). Standardized discs and 1:1, 1:2, and 1:4 eluates of the tested materials were prepared. The following in vitro experiments were carried out: ion release, cell metabolic activity 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell migration, immunofluorescence experiment, cell attachment, gene expression, and mineralization assay. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p < .05). Increased Ca 2+ release was detected in TFbc compared to AHPbc and Neo (*p < .05). Biological assays showed a discrete cell metabolic activity and cell migration in Neo-treated cell, whereas scanning electronic microscopy assay exhibited that TFbc group had a better cell adhesion process of substrate attachment, spreading, and cytoskeleton development on the niche-like structures of the cement than AHPbc and Neo. The sealers tested were able to induce overexpression of the CEMP-1, ALP, and COL1A1 genes in the first days of exposure, particularly in the case of TFbc (***p < .001). All materials tested significantly increased the mineralization of hPDLSCs when compared to the negative control, although more pronounced calcium deposition was observed in the TFbc-treated cells (***p < .001). Our results suggested that TFbc promotes cell differentiation, both by increasing the expression of key osteo/odontogenic genes and by promoting mineralization of the extracellular matrix, whereas this phenomenon was less evident in Neo and AHPbc. RESEARCH HIGHLIGHTS: TFbc group had a better cell adhesion process of substrate attachment, spreading, and cytoskeleton development on the niche-like structures of the cement than AHPbc and Neo. The sealers tested were able to induce overexpression of the CEMP-1, ALP, and COL1A1 genes in the first days of exposure, particularly in the case of TFbc. All materials tested significantly increased the mineralization of hPDLSCs when compared to the negative control, although more pronounced calcium deposition was observed in the TFbc-treated cells., (© 2024 The Authors. Microscopy Research and Technique published by Wiley Periodicals LLC.)

Introduction: BioRoot Flow (BRF) is a novel premixed bioceramic sealer indicated for endodontic treatments, but the biological and immunomodulatory effects of this endodontic sealer on human periodontal ligament stem cells (hPDLSCs) have not been elucidated., Methods: To ascertain the biological impact of BRF, TotalFill BC Sealer (TFbc), and AH Plus (AHP) on human Periodontal Ligament Stem Cells (hPDLSCs), assessments were conducted to evaluate the cytocompatibility, cellular proliferation, migratory capacity, osteo/cementogenic differentiation potential, the ability to form mineralized nodules, and the immunomodulatory characteristics of hPDLSCs following treatment with these endodontic sealers., Results: Biological assays showed adequate cell metabolic activity and cell migration in BRF, while SEM assay evidenced that TFbc and BRF groups demonstrated a superior cell adhesion process, including substrate adhesion, cytoskeleton development, and spreading on the niche-like structures of the cement as compared to the AHP group. TFbc and BRF-treated groups exhibited a significantly lower IL6 and IL8 production than AHP (* p <.05). The bioceramic sealers stimulated heightened expression of BSP, CEMP-1, and CAP genes within a 7-14 day period. Notably, BRF and TFbc demonstrated a significant enhancement in the mineralization of hPDLSCs when compared to the negative control. Among these, cells treated with BRF showed a more substantial accumulation of calcium (*** p < .001)., Conclusions: Taken together, these findings indicate that BRF can potentially enhance cell differentiation by promoting the expression of essential genes related to bone and cement formation. In addition, BRF and TFbc displayed anti-inflammatory effects., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Introduction: nanoparticles are tiny-sized materials whose characteristics and properties mean that their association with dental materials is being investigated to ascertain their effects and possible benefits on tooth structures. This systematic review aimed to qualitatively collect in vitro studies that address the potential application of different nanoparticles in dental regeneration. Following an exhaustive search and article selection process, 16 in vitro studies that met our eligibility criteria were included. BG-NPs were analyzed across five studies, with three demonstrating their impact on the growth and differentiation of human hDPSCs. CS-NPs were examined in three studies, with findings from two indicating a significant effect on the differentiation of SCAPs. Nanoparticles' therapeutic potential and their stimulatory effect on promoting the regeneration of cells of the dentin-pulp complex have been proven. Their effect is altered according to the type of nanoparticle, concentration, and substances associated with them and, depending on these variables, they will affect the pulp, dentine, and dental cementum differently., Competing Interests: The authors declare no conflicts of interest related to this article.

The aim of this study was to assess the influence of eucalyptol and menthol on the cell viability, migration, and reactive oxygen species production of human gingival fibroblasts (GFs) in vitro. Three different concentrations of eucalyptol and menthol were prepared following ISO 10993-5 guidelines (1, 5, and 10 mM). GFs were isolated from extracted teeth from healthy donors. The following parameters were assessed: cell viability via MTT, Annexin-V-FITC and 7-AAD staining, and IC 50 assays; cell migration via horizontal scratch wound assay; and cell oxidative stress via reactive oxygen species assay. Data were analyzed using one-way ANOVA and Tukey's post hoc test. Statistical significance was established at p < 0.05. Eucalyptol and Menthol exhibited high cytotoxicity on gingival fibroblasts, as evidenced by cytotoxicity assays. Eucalyptol showed lower levels of cytotoxicity than menthol, compared to the control group. The cytotoxicity of the tested substances increased in a concentration-dependent manner. The same occurred in a time-dependent manner, although even 10 min of exposure to the tested substances showed a high cytotoxicity to the GFs. Commercially available products for oral application with these substances in their composition should be tested for cytotoxicity before their use.

Objectives: The current in vitro study aims to evaluate silk fibroin with and without the addition of graphene as a potential scaffold material for regenerative endodontics., Material and Methods: Silk fibroin (SF), Silk fibroin/graphene oxide (SF/GO) and silk fibroin coated with reduced graphene oxide (SF/rGO) scaffolds were prepared (n = 30). The microarchitectures and mechanical properties of scaffolds were evaluated using field emission scanning electron microscopy (FESEM), pore size and water uptake, attenuated total reflectance fourier transformed infrared spectroscopy (ATR-FTIR), Raman spectroscopy and mechanical compression tests. Next, the study analyzed the influence of these scaffolds on human dental pulp stem cell (hDPSC) viability, apoptosis or necrosis, cell adhesion, odontogenic differentiation marker expression and mineralized matrix deposition. The data were analyzed with ANOVA complemented with the Tukey post-hoc test (p < 0.005)., Results: SEM analysis revealed abundant pores with a size greater than 50 nm on the surface of tested scaffolds, primarily between 50 nm and 600 µm. The average value of water uptake obtained in pure fibroin scaffolds was statistically higher than that of those containing GO or rGO (p < 0.05). ATR-FTIR evidenced that the secondary structures did not present differences between pure fibroin and fibroin coated with graphene oxide, with a similar infrared spectrum in all tested scaffolds. Raman spectroscopy showed a greater number of defects in the links in SF/rGO scaffolds due to the reduction of graphene. In addition, adequate mechanical properties were exhibited by the tested scaffolds. Regarding biological properties, hDPSCs attached to scaffolds were capable of proliferating at a rate similar to the control, without affecting their viability over time. A significant upregulation of ALP, ON and DSPP markers was observed with SF/rGO and SF/GO groups. Finally, SF/GO and SF/rGO promoted a significantly higher mineralization than the control at 21 days., Significance: Data obtained suggested that SF/GO and SF/rGO scaffolds promote hDPSC differentiation at a genetic level, increasing the expression of key osteo/odontogenic markers, and supports the mineralization of the extracellular matrix. However, results from this study are to be interpreted with caution, requiring further in vivo studies to confirm the potential of these scaffolds., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Objectives: Silver fluoride (SF) is a preventive and therapeutic option for dental pathological processes involving structural alterations of the hard tissues, either during their formation or those caused by caries or other pathological reasons. This study aimed to compare the biological properties of two commercial SF products, one of them with ammonium (Riva Star; SDF) and the other ammonium-free (Riva Star Aqua; AgF), both with or without potassium iodide (KI), by the assessment of the cytotoxicity of the materials' eluates., Methods: Human dental pulp stem cells (hDPSCs) were obtained from healthy 18-23-year-old donors. Three dilutions were prepared for the tested materials (0.005%, 0.0005%, and 0.0001%). The following groups were assessed: (AgF, AgF+KI, SDF, SDF+KI, KI, negative control). A series of cytocompatibility assays were performed: MTT assay, IC50 assay, wound healing (migration) assay, cell cytoskeleton staining, analysis of cell apoptosis and necrosis, and reactive oxygen species production. The normality in the distribution of the data was previously confirmed via a Q-Q plot. Statistical significance was tested using one way ANOVA and Tukey's post hoc test., Results: The incorporation of KI improved the cytocompatibility of both SF products in terms of viability, migration, morphology, apoptosis, and reactive oxygen species production. This difference was higher in the AgF group. The lowest dilutions of SF+KI and AgF+KI showed a similar cytocompatibility to that of the control group (MTT assay (p > 0.05 after 24, 48, and 72 h of culture); migration assay (p > 0.05 after 24, 48, and 72 h of culture); reactive oxygen species production (p > 0.05 after 72 h of culture)., Significance: Riva Star Aqua shows lower cytotoxicity than Riva Star on hDPSCs. It can be considered as a good alternative in the conservative treatment of dental caries and in the preservation and remineralisation of viable dentine tissue., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: The period of pregnancy is characterized by a multitude of diverse changes that exert a notable impact on the oral cavity of women. During this gestational phase, patients necessitate tailored oral care and specific recommendations to preempt and address potential oral diseases. This systematic review aimed to perform a detailed analysis of the research studies that focused on the oral manifestations observed in pregnant women. Methods: A meticulous search was conducted in the databases Medline, Scopus, and Scielo by employing the following search terms: ((pregnant OR pregnancy)) AND (("oral manifestation*") OR ("oral health")). Articles that were published between 2013 and 2023 and written in English or Spanish and studies that scrutinized oral manifestations in pregnant women and included a diagnosis conducted by a qualified dentist were selected; we excluded articles published before 2013, articles that could not be accessed in full text, studies whose patients were not pregnant women at the time of the study, studies where patients were selected because they had a specific disease, studies where the clinical examination was not performed by a dentist, and articles written in languages other than English or Spanish. Subsequently, the risk of bias in the chosen articles was assessed in accordance with the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) scale. Results: A total of 20 studies were included in the analysis, following the exclusion and inclusion criteria. These studies were categorized as cross-sectional, cohort, longitudinal, or case-control. Various oral manifestations in pregnant women were examined, with five studies comparing these manifestations with those observed in nonpregnant women. Conclusions: The most prevalent oral manifestations associated with pregnancy encompass dental caries, periodontitis, gingivitis, pyogenic granuloma, and candidiasis. Nonetheless, less common lesions may also emerge during the course of pregnancy.

Crohn's disease (CD) is a chronic inflammatory intestinal condition that can affect the entire gastrointestinal tract. It is characterized by its clinical heterogeneity and irregularities in its course. The etiology and pathogenesis are not well established, so it is difficult to establish an early diagnosis and an effective treatment plan. The objective of this systematic review was to present a qualitative synthesis of the studies referring to the oral manifestations of CD. This systematic review was carried out following the PRISMA guide. Research was conducted in the Pubmed, Web of Science, Scopus, Scielo, and Cocrahne Library databases on 23 February 2023, and updated on 1 September 2023. Articles published between 2012 and 2023 were selected. Articles that analyzed the oral manifestation of CD patients and met the established search terms. In addition, the quality of all the selected studies was analyzed following the CARE guidelines for case reports and the STROBE scale for observational studies. A total of 19 articles were included in this review that met the inclusion criteria. Regarding the oral manifestation of CD, oral ulcers, angular cheilitis, and gingivitis stand out. Periodontitis and vegetative pyostomatitis were the least representative manifestations. The most prevalent locations were lips, mucosa, and gingivae. Ulcers, gingivitis, and angular cheilitis are the most frequent oral manifestations in patients with CD. Their early identification and possible relationship with the disease are important for an early diagnosis and an adequate treatment plan.

Objectives: To assess the cytocompatibility and bioactive potential of the new calcium silicate-based cement Ceraputty on human periodontal ligament stem cells (hPDLSCs) compared to Biodentine and Endosequence BC root repair material (ERRM)., Materials and Methods: hPDLSCs were isolated from extracted third molars from healthy donors. Standardized sample discs and 1:1, 1:2, and 1:4 eluates of the tested materials were prepared. The following assays were performed: surface element distribution via SEM-EDX, cell attachment and morphology via SEM, cell viability via a MTT assay, osteo/cemento/odontogenic marker expression via RT-qPCR, and cell calcified nodule formation via Alizarin Red S staining. hPDLSCs cultured in unconditioned or osteogenic media were used as negative and positive control groups, respectively. Statistical analysis was performed using one-way ANOVA or two-way ANOVA and Tukey's post hoc test. Statistical significance was established at p < 0.05., Results: The highest Ca 2+ peak was detected from Biodentine samples, followed by ERRM and Ceraputty. hPDLSC viability was significantly reduced in Ceraputty samples (p < 0.001), while 1:2 and 1:4 Biodentine and ERRM samples similar results to that of the negative control (p > 0.05). Biodentine and ERRM exhibited an upregulation of at least one cemento/odonto/osteogenic marker compared to the negative and positive control groups. Cells cultured with Biodentine produced a significantly higher calcified nodule formation than ERRM and Ceraputty (p < 0.001), which were also higher than the control groups (p < 0.001)., Conclusion: Ceraputty evidenced a reduced cytocompatibility towards hPDLSCs on its lowest dilutions compared to the other tested cements and the control group. Biodentine and ERRM promoted a significantly higher mineralization and osteo/cementogenic marker expression on hPDLSCs compared with Ceraputty. Further studies are necessary to verify the biological properties of this new material and its adequacy as a retrograde filling material., Clinical Relevance: This is the first study to elucidate the adequate biological properties of Ceraputty for its use as a retrograde filling material., (© 2023. The Author(s).)

In the last few decades, biomimetic concepts have been widely adopted in various biomedical fields, including clinical dentistry. Endodontics is an important sub-branch of dentistry which deals with the different conditions of pulp to prevent tooth loss. Traditionally, common procedures, namely pulp capping, root canal treatment, apexification, and apexigonesis, have been considered for the treatment of different pulp conditions using selected materials. However, clinically to regenerate dental pulp, tissue engineering has been advocated as a feasible approach. Currently, new trends are emerging in terms of regenerative endodontics which have led to the replacement of diseased and non-vital teeth into the functional and healthy dentine-pulp complex. Root- canal therapy is the standard management option when dental pulp is damaged irreversibly. This treatment modality involves soft-tissue removal and then filling that gap through the obturation technique with a synthetic material. The formation of tubular dentine and pulp-like tissue formation occurs when stem cells are transplanted into the root canal with an appropriate scaffold material. To sum up tissue engineering approach includes three components: (1) scaffold, (2) differentiation, growth, and factors, and (3) the recruitment of stem cells within the pulp or from the periapical region. The aim of this paper is to thoroughly review and discuss various pulp-regenerative approaches and materials used in regenerative endodontics which may highlight the current trends and future research prospects in this particular area., Competing Interests: The authors declare no conflict of interest.

The aim of this study was to assess the influence of eucalyptol, chloroform, and Endosolv on the proliferative capability, cell viability, and migration rates of human periodontal ligament stem cells (hPDLSCs) in vitro. Solvent eluates were formulated following ISO 10993-5 guidelines, and 1%, 0.25%, and 0.1% dilutions were prepared. The HPDLSCs were isolated from the extracted third molars of healthy donors. The following parameters were assessed: cell viability via trypan blue and IC50 assays, cell migration via horizontal wound healing assay, cell morphology via cell cytoskeleton staining (phalloidin labeling), and cell oxidative stress via reactive oxygen species assay. The data were analyzed using one-way ANOVA and Tukey’s posthoc tests, and their significance was established at p < 0.05. Chloroform and eucalyptol exhibited significantly higher cytotoxicity on the hPDLSCs in vitro compared to the control group, as shown by the cell viability, migration, morphology, and reactive oxygen species release assays. Alternatively, Endosolv showed adequate cytotoxicity levels comparable to those of the control group. The cytotoxicity of the tested endodontic solvents increased in a dose-dependent manner. The results from the present study highlight the cytotoxicity of chloroform and eucalyptol. Thus, their limited and cautious use is recommended, avoiding solvent extrusion.

The main objective of this paper is to perform an updated literature review of guided endodontics based on the available up-to-date scientific literature to identify and describe the technique, its benefits, and its limitations. Four electronic databases (PubMed, Scopus, Science Direct, and Web of Science) were used to perform a literature search from 1 January 2017 to 13 May 2022. After discarding duplicates, out of 1047 results, a total of 29 articles were eligible for review. Guided endodontics is a novel technique that is currently evolving. It is applied in multiple treatments, especially in accessing and locating root canals in teeth with pulp canal obliteration, microsurgical endodontics, and removing glass fiber posts in endodontic retreatments. In addition, it is independent of an operator's experience, requires less treatment time for the patient, and is more accurate and safer than conventional endodontics.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that is characterized by clinical heterogeneity and irregularities in its course. The etiology and pathogenesis of this pathology are not well-understood, so there is difficulty in establishing a diagnosis and treatment plan with certainty. The aim of this systematic review is to present a qualitative synthesis of studies referring to the oral manifestations of systemic lupus erythematosus (SLE). This systematic review was performed following the PRISMA guideline. On this basis, a search for articles was performed in the PubMed, Web of Science, and Scopus databases on 19 November 2021 and updated on 15 February 2022. We chose articles published between 2012 and 2022 that analyzed the oral manifestations of SLE patients. The quality of all these studies was analyzed following the STROBE scale. A total of 15 articles were included in this study after selection. The selected articles were cross-sectional, case-control, and cohort studies. The most frequently associated oral manifestations with SLE were oral ulcers, hyposalivation, pigmentations, glossodynia, cleft tongue, cheilitis, arthritis, and secondary Sjögren's syndrome. However, despite the importance of the perception of these oral manifestations in the early diagnosis of SLE, there are still not enough studies about them.

Background: Bleeding complications in patients undergoing antiplatelet and/or anticoagulant therapy have been one of the main concerns in dental practice. Upon the introduction of new antiplatelet and anticoagulant drugs, there is a search for new protocols that respond to a secure treatment. The aim of the present study was to evaluate bleeding complications in anticoagulated and antiplatelet-treated patients after performing simple dental extractions, in a period of 4 years., Material and Methods: 147 clinical records of anticoagulated and/or antiplatelet-treated patients undergoing a simple dental extraction over a period of 4 years (October 2015 to September 2019) were studied. Within the sample, 63 patients were antiplatelet-treated, 83 were anticoagulated, and 1 patient was under both therapies. Within the anticoagulated patients, 70 took classic anticoagulants and 14 new oral anticoagulants (NOACs). Quantitative data were studied with arithmetic mean and standard deviation (SD). The chi-square test was used for the qualitative variables. ANOVA tests were used to compare age and anticoagulated or antiplatelet-treated patients. Statistical significance was determined when p < 0.05., Results: From the 418 dental extractions performed, five severe bleeding complications took place in three patients (2.11%). From the five events, four were in patients treated with NOACs (1.68%) and one occurred in a patient anticoagulated with acenocoumarol (0.42%; p = 0.003)., Conclusions: Considering the results of this retrospective clinical study, we can conclude that bleeding complications in anticoagulated and/or antiplatelet-treated patients after tooth extractions were low, with a higher incidence recorded in patients treated with NOACs, followed by classic anticoagulants, and there were no complications in antiplatelet-treated patients.

The purpose is to analyze the medical characteristics of children with special health care needs (CSHCN) recommended for dental treatment under general anesthesia (GA), postoperative complications, and dental treatment outcomes under the regulation of the Spanish Dental Care Program (PADI). 111 clinical records were selected. The study population was divided into three age groups. The quantitative data was specified as the mean ± SD. For the qualitative variables, the Chi-Square test was used. One-way ANOVA and Bonferroni tests were used to examine the effect of the "age group" and the number of treatment procedures. A total of 1473 treatment procedures were performed, of which 110 (7.5%) were cleanings, 898 (61%) were restorative procedures, 332 (21.7%) were extractions, 22 (1.6%) were endodontic treatments, 62 (4.2%) were pulpotomies, and 59 (4%) were stainless steel crowns. Regarding the mean number of incisor root canal treatments (RCT), age group 3 received a significantly higher mean number of incisor RCTs than age group 1 ( p = 0.02). Age group 1 received a higher average of pulpotomies and stainless-steel crowns ( p = 0.00) compared to groups 2 and 3. GA is a safe procedure for the dental treatment of CSHCN, with minimal postoperative complications, which should be included among dental public programs.

Objectives: To evaluate the possible synergic effect of cisplatin and low molecular weight heparin (LMWH) on oral squamous cell carcinoma (OSCC)., Materials and Methods: Cisplatin and enoxaparin sodium, alone or in combination, were administered at doses of 1, 2, 4, 8 and 10 µM and 0.1, 0.5, 1, 5, 10, 50, and 100 µg/ml, respectively, to the H357 human OSCC line. The effects on cell viability and apoptosis were evaluated after 24, 48, and 72 h and on cell migration after 18 and 24 h., Results: 10 µM concentration of cisplatin produced the greatest decrease in cell viability, with significant differences at 24 ( p =0.009), 48 ( p =0.001) and 72 h ( p = 0.003); the 100 µg/ml dose of enoxaparin produced the greatest decrease in cell viability but without significant differences ( p >0.05). When different concentrations of cisplatin and enoxaparin were combined, it was found that 100 µg/ml enoxaparin sodium produced the greatest synergic effect on cell viability reduction. In analyses of apoptosis and cell migration, it was found that the combination of cisplatin at 8 or 10 μM and 100 μg/ml enoxaparin produced a higher rate of apoptosis at 24, 48, and 72 h and a greater reduction in cell migration at 18 and 24 h., Conclusions: A combination of cisplatin and enoxaparin sodium shows a synergic effect that reduces cell viability and cell migration capacity and increases the apoptosis of human OSCC cells., Clinical Relevance: Enoxaparin may be beneficial in chemotherapy for patients with OSCC; this finding requires further clinical and laboratory investigation., (Copyright © 2020 Camacho-Alonso, Gómez-Albentosa, Oñate-Sánchez, Tudela-Mulero, Sánchez-Siles, Gómez-García and Guerrero-Sánchez.)

This study aimed to analyze the biological effects of three new bioactive materials on cell survival, migration, morphology, and attachment in vitro. ACTIVA Kids BioACTIVE Restorative (Pulpdent, Watertown, MA, USA) (Activa), Ionolux (Voco, Cuxhaven, Germany), and Riva Light Cure UV (SDI, Bayswater, Australia) (Riva) were handled and conditioned with a serum-free culture medium. Stem cells from human dental pulp (hDPSCs) were exposed to material extracts, and metabolic activity, cell migration, and cell morphology were evaluated. Cell adhesion to the different materials was analyzed by scanning electron microscopy (SEM). The chemical composition of the materials was evaluated by energy-dispersive X-ray (EDX). One-way analysis of variance followed by a Tukey test was performed ( p < 0.05). Ionolux promoted a drastic reduction in metabolic activity and wound closure compared to the control ( p < 0.05), whereas Activa induced adequate metabolic activity and cell migration. Moreover, SEM and immunofluorescence analysis showed abundant cells exposed to Activa. The materials showed different surface morphologies, and EDX spectra exhibited different peaks of C, O, Si, S, Ca, and F ions in glass ionomer cements. The results showed that Activa induced cell migration, cell attachment, and cell viability to a greater extent than Riva and Ionolux.

Objective: The aim of this study was to analyze the biological effects of MTA Repair HP and ProRoot MTA on human periodontal ligament stem cells (hPDLSCs) after exposure to acidic and neutral environments., Materials and Methods: Discs of each material (n = 30) were exposed to phosphate buffered saline (pH = 7.4) or butyric acid (pH = 5.2) for 7 days, and biological testing was carried out in vitro on hPDLSCs. Cell viability and apoptosis assays were performed using eluates of each root-end filling material. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. The chemical composition of the root-end filling materials was determined by energy-dispersive x-ray and eluates were analyzed by inductively coupled plasma-mass spectrometry. Statistical differences were assessed by ANOVA and Tukey test (p < 0.05)., Results: Under an acidic environment, both materials displayed similar ion release abilities, with the increased release of Si and Ca ions. Substantial changes in microstructure were observed for both materials after exposure to acidic pH. In addition, material exposure to an acidic environment showed a similar degree of cell adherence, and, surprisingly, MTA Repair HP exhibited higher cell viability rates at pH 5.2 than ProRoot MTA., Conclusions: Exposure to an acidic environment promoted Si and Ca ion release from ProRoot MTA and MTA Repair HP. Moreover, we observed optimal biological properties of ProRoot MTA and MTA Repair HP in terms of cell viability, cell death, and cell attachment in both environments., Clinical Relevance: These results may suggest that MTA Repair HP and ProRoot exhibited optimal biological properties in terms of cell viability, cell death and cell attachment in acidic environment, being considered as materials for root-end filling and perforations.

Aim: To analyse in vitro changes in ion release and biological properties of Endocem-MTA (Maruchi, Wonju, Korea) and NeoMTA-Plus (Avalon Biomed Inc, Bradenton, FL, USA) exposed to acidic or neutral environment on human dental periodontal ligament stem cells (hPDLSCs)., Methodology: Cell viability and wound healing assays were performed using eluates of each material. Cell death and changes in phenotype induced by the set endodontic sealer eluates were evaluated through flow cytometry. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy. The chemical composition of the materials was determined by energy-dispersive X-ray (EDX), and ion release was evaluated by inductively coupled plasma-mass spectrometry. Statistical analysis was performed with analysis of variance and a Bonferroni or Tukey post-test (α < 0.05)., Results: The MTT assay revealed non-cytotoxic effects of NeoMTA-Plus and Endocem-MTA at pH 5.2 and 7.4. However, there were minor differences compared with the control, especially at pH 5.2, where both materials were associated with significantly greater cell viability (P < 0.05). In both environments, the materials stimulated hPDLSCs to migrate. hPDLSCs were attached to the bioactive cements, with multiple prolongations proliferated on the surface of the samples. Moreover, there were no changes to cell phenotype or apoptosis/necrosis rates, indicating that the acidic environment did not induce cell death. Prismatic crystalline structures were seen on the surface of the cements exposed to butyric acid and EDX analysis identified a marked peak of Ca 2+ from NeoMTA-Plus and Endocem-MTA in acidic and physiological environments., Conclusions: An acidic environment favoured the release of Ca 2+ ions from both bioactive cements, and the cytotoxicity of these bioactive cements was low in both environments studied., (© 2019 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Multiple side effects related to bleaching were found to occur in the dental pulp tissue, including decreased cell metabolism and viability. In this work we evaluated the in vitro diffusion capacity, cytotoxicity and biocompatibility of four commercial bleaching products on stem cells from human dental pulp (hDPSCs). Two commercial bleaching gels hydrogen peroxide-based (HP), Norblanc Office 37.5% (Nor-HP) and Opalescence Boost 40% (Opal-HP) were applied for 30 min to enamel/dentine discs. Another two gels from the same manufacturers, 16% carbamide peroxide-based (CP), Norblanc Home (Nor-CP) and Opalescence CP 16% (Opal-CP), were applied for 90 min. The diffusion of HP was analysed by fluorometry. Cytotoxicity was determined using the MTT assays, the determination of apoptosis, immunofluorescence assays and intracellular reactive oxygen species (ROS) level. Tissue inflammatory reactions were evaluated histopathologically in rats. Statistical differences were performed by one-way ANOVA and Bonferroni post-test (α < 0.05). Normon products showed lower cytotoxicity and diffusion capacity than the Ultradent products. A high intracellular ROS level was measured in hDPSCs after exposure to Opal-HP. Finally, a severe necrosis of both coronal and radicular pulp was observed with Opal-HP. Similar concentrations of hydrogen peroxide and carbamide peroxide in a variety of bleaching products exhibited different responses in cells and dental pulp tissue, suggesting that bleaching products contain unknown agents that could influence their toxicity.

The purpose of this work was to evaluate the physicochemical properties, the cytotoxicity and in vivo biocompatibility of MTA Repair HP (MTA HP) and White MTA (WMTA). The setting time, flow, radiopacity and water solubility were assessed. To the cytotoxicity assay, primary human osteoblast cells were exposed to several dilutions of both materials eluates. MTT assay, apoptosis assay and cell adhesion assay were performed. The in vivo biocompatibility was evaluated through histological analysis using different staining techniques. No differences were observed between MTA HP and WMTA for setting time, radiopacity, solubility and water absorption (P > 0.05). However, MTA HP showed a significantly higher flow when compared to WMTA (P < 0.05). Cell viability results revealed that the extracts of WMTA and MTA HP promoted the viability of osteoblasts. After incubation of cells with the endodontic cement extracts, the percentage of apoptotic or necrotic cells was very low (<3%). Furthermore, SEM results showed a high degree of cell proliferation and adhesion on both groups. MTA HP showed similar in vivo biocompatibility to the WMTA and the control group in all time-points. The MTA HP presented adequate physicochemical and biological properties with improved flow ability when compared to WMTA. Such improved flow ability may be a result of the addition of a plasticizing agent and should be related to an improvement in the handling of MTA HP.

Objectives: To evaluate in vitro the cementogenic potential and the biological effects of GuttaFlow Bioseal, GuttaFlow 2, MTA Fillapex and AH Plus on human periodontal ligament stem cells (hPDLSCs)., Methods: Cell viability, cell migration and cell morphology assays were performed using eluates of each material. To evaluate cell attachment, hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy (SEM). The effects of endodontic sealers on cementum protein 1 (CEMP1), cementum-derived attachment protein (CAP), bone sialoprotein (BSP), ameloblastin (AMBN), amelogenin (AMELX) and alkaline phosphatase (ALP) gene expression on hPDLSCs were investigated by qPCR and immunofluorescence (IF). Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α<0.05)., Results: More than 90% of viable cells were obtained using extracts of GuttaFlow Bioseal and GuttaFlow2 after 72h of culture. By contrast, AH Plus and MTA Fillapex induced significantly lower levels of cell viability. GuttaFlow2 and GuttaFlow Bioseal promoted wound closure in a concentration-dependent manner, comparable to that observed with control extracts (*p<0.05). However, with AH Plus and MTA Fillapex, cell migration was significantly lower than in the control (***p<0.0001). SEM analysis pointed to an organized stress fiber assembly and high degree of cell adhesion on GuttaFlow Bioseal disks but low rates on GuttaFlow2, MTA Fillapex and AH Plus. When hPDLSCs were cultured with GuttaFlow Bioseal-conditioned media, qPCR assays and IF showed a higher level of AMELX, AMBN, CEMP1 and CAP expression than the control (*p<0.05)), whereas no such expression was observed in the other sealers., Significance: Our results showed that GuttaFlow sealers were more cytocompatible than AH Plus and MTA Fillapex, while GuttaFlow Bioseal favored cementoblast differentiation of hPDLSCs in the absence of any growth factors., (Copyright © 2018 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

Background : Cell-Based Therapies (CBT) constitute a valid procedure for increasing the quantity and quality of bone in areas with an inadequate bone volume. However, safety and efficacy should be investigated prior to clinical application. The objective of this study was to evaluate the biodistribution, safety and osteogenic capacity of bone marrow-derived human mesenchymal stem cells ( h BMMSCs) pre-seeded into β-tricalcium phosphate (TCP) and implanted into NOD/SCID mice at subcutaneous and intramuscular sites. Methods : h BMMSCs were isolated, characterized and then cultured in vitro on a porous β-TCP scaffold. Cell viability and attachment were analyzed and then h BMMSCs seeded constructs were surgically placed at subcutaneous and intramuscular dorsal sites into NOD/SCID mice. Acute and subchronic toxicity, cell biodistribution and efficacy were investigated. Results : There were no deaths or adverse events in treated mice during the 48-hour observation period, and no toxic response was observed in mice. In the 12-week subchronic toxicity study, no mortalities, abnormal behavioral symptoms or clinical signs were observed in the saline control mice or the h BMMSCs/β-TCP groups. Finally, our results showed the bone-forming capacity of h BMMSCs/β-TCP since immunohistochemical expression of human osteocalcin was detected from week 7. Conclusions : These results show that transplantation of h BMMSCs/β-TCP in NOD/SCID mice are safe and effective, and might be applied to human bone diseases in future clinical trials.

The purpose of this study was to evaluate the diffusion capacity and the biological effects of different bleaching products on human dental pulp stem cells ( h DPSCs). The bleaching gel was applied for 90, 30 or 15 min to enamel/dentine discs that adapted in an artificial chamber. The diffusion of hydrogen peroxide (HP) was analysed by fluorometry and the diffusion products were applied to h DPSCs. Cell viability, cell migration and cell morphology assays were performed using the eluates of diffusion products. Finally, cell apoptosis and the expression of mesenchymal stem cell markers were analysed by flow cytometry. Statistical analysis was performed using analysis of variance and Kruskal⁻Wallis or Mann⁻Whitney tests (&alpha; < 0.05). Significant reductions of approximately 95% in cell viability were observed for the 3 &times; 15 min groups ( p < 0.001), while 1 &times; 30 min of PerfectBleach and 1 &times; 90 min of PolaNight resulted in reductions of 50% and 60% in cell viability, respectively ( p < 0.001). Similar results were obtained in the migration assay. Moreover, the 3 &times; 15 min group was associated with cell morphology alterations and reductions of >70% in cell live. Finally, h DPSCs maintained their mesenchymal phenotype in all conditions. Similar concentrations of carbamide peroxide (CP) and HP in different commercial products exhibited different biological effects on hDPSCs.

Objective: To evaluate the in vitro cytotoxicity of Equia Forte (GC, Tokyo, Japan) and Ionostar Molar (Voco, Cuxhaven, Germany) on human dental pulp stem cells (hDPSCs)., Methods: hDPSCs isolated from third molars were exposed to several dilutions of Equia Forte and Ionostar Molar eluates (1/1, 1/2 and 1/4). These eluates were obtained by storing material samples in respective cell culture medium for 24h (n=40). hDPSCs in basal growth culture medium were the control. Cell viability and cell migration assays were performed using the MTT and wound-healing assays, respectively. Also, induction of apoptosis and changes in cell phenotype were evaluated by flow cytometry. Changes in cell morphology were analysed by immunocytofluorescence staining. To evaluate cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy (SEM). The chemical composition of the materials was determined by energy dispersive X-ray (EDX) and eluates were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS). Statistical analysis was performed with analysis of variance (ANOVA) and Student's t-test (α<0.05)., Results: Undiluted Equia Forte extracts led to a similar cell proliferation rates than the control group from 72h onwards. There were no significance differences between Equia Forte and Ionostar Molar in terms of cell apoptosis and phenotype. However, in presence of Equia extracts the migration capacity of hDPSCs was higher than in presence of Ionostar Molar (p<0.05). Also, SEM studies showed a higher degree of cell attachment when Equia Forte extracts were used. Finally, EDX analysis pointed to different weight percentages of C, O and Ca ions in glass ionomer cements, while other elements such as La, Al, Si, W, Mo and F were also detected., Significance: In summary, Equia Forte promoted better biological responses in hDPSCs than Ionostar Molar., (Copyright © 2018 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

The aim is to investigate in vitro biological effects of silk fibroin 3D scaffolds on stem cells from human exfoliated deciduous teeth (SHEDs) in terms of proliferation, morphological appearance, cell viability, and expression of mesenchymal stem cell markers. Silk fibroin 3D scaffolding materials may represent promising suitable scaffolds for their application in regenerative endodontic therapy approaches. SHEDs were cultured in silk fibroin 3D scaffolds. Then, cell numbers were counted and the Alamar blue colorimetric assay was used to analyse cell proliferation after 24, 48, 72, and 168 h of culture. The morphological features of SHEDs cultured on silk fibroin scaffolds were evaluated by scanning electron microscopy (SEM). Finally, cell viability and the expression of mesenchymal stem cell markers were analysed by flow cytometry. One-way analysis of variance (ANOVA) followed by a Bonferroni post-test was performed (P < 0.05). At 24 and 48 h of culture, SHED proliferation on scaffolds was modest compared to the control although still significant (p < 0.05). However, cell proliferation progressively increased from 72 to 168 h compared with the control (p < 0.001; p < 0.01). In addition, flow cytometry analysis showed that the culture of SHEDs on silk fibroin scaffolds did not significantly alter the level of expression of the mesenchymal markers CD73, CD90, or CD105 up to 168 h; in addition, cell viability in silk fibroin was similar to than obtained in plastic. Moreover, SEM studies revealed a suitable degree of proliferation, cell spreading, and attachment, especially after 168 h of culture. The findings from the current study suggest that silk fibroin 3D scaffolds had a favourable effect on the biological responses of SHEDs. Further in vivo investigations are required to confirm these results.

Introduction: The aim of the present study was to evaluate the in vitro cytotoxicity of MTA Repair HP, NeoMTA Plus, and Biodentine, new bioactive materials used for dental pulp capping, on human dental pulp stem cells (hDPSCs)., Methods: Biological testing was carried out in vitro on hDPSCs. Cell viability and cell migration assays were performed using eluates of each capping material. To evaluate cell morphology and cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. The chemical composition of the pulp-capping materials was determined by energy-dispersive X-ray and eluates were analyzed by inductively coupled plasma-mass spectrometry. Statistical differences were assessed by analysis of variance and Tukey test (P < .05)., Results: Cell viability was moderate after 24 and 48 hours in the presence of MTA Repair HP and NeoMTA Plus, whereas at 48 and 72 hours, Biodentine showed higher rates of cell viability than MTA Repair HP and NeoMTA Plus (P < .001). A cell migration assay revealed adequate cell migration rates for MTA Repair HP and NeoMTA Plus, both similar to the control group rates, meanwhile the highest cell migration rate was observed in the presence of Biodentine (P < .001). Scanning electron microscope studies showed a high degree of cell proliferation and adhesion on Biodentine disks but moderate rates on MTA Repair HP and NeoMTA Plus disks. Energy-dispersive X-ray pointed to similar weight percentages of C, O, and Ca in all 3 materials, whereas other elements such as Al, Si, and S were also found., Conclusions: The new pulp-capping materials MTA Repair HP, NeoMTA Plus, and Biodentine showed a suitable degree of cytocompatibility with hDPSCs, and good cell migration rates, although Biodentine showed higher rates of proliferation time-dependent., (Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Aim: To evaluate the biological effects in vitro of MTA-Angelus (MTA-Ang; Angelus, Londrina, PR, Brazil), MTA Repair HP (MTA-HP; Angelus) and NeoMTA Plus (NeoMTA-P; Avalon Biomed Inc, Bradenton, FL, USA) on human dental pulp stem cells (hDPSCs)., Methodology: Cell viability and cell migration assays were performed using eluates of each material. To evaluate cell morphology and cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analysed by immunocytofluorescence and scanning electron microscopy, respectively. The chemical composition of the materials was determined by energy-dispersive X-ray (EDX), and eluates were analysed by inductively coupled plasma-mass spectrometry (ICP-MS). Statistical analysis was performed with the analysis of variance and Bonferroni or Tukey post-test (α < 0.05)., Results: Undiluted MTA-Ang, MTA-HP and NeoMTA-P displayed a significant increase in cell viability greater than that obtained using complete medium alone (control) (*P < 0.05; **P < 0.01; ***P < 0.001). Moreover, a cell migration assay revealed cell migration rates after incubation with extracts of MTA-Ang, MTA-HP and NeoMTA-P that were similar to levels obtained in the control group. In addition, stretched cytoskeletal F-actin fibres were detected in the cells treated with the three material extracts. SEM studies revealed a high degree of cell proliferation and attachment on all three materials. EDX analysis demonstrated similar weight percentages of C, O and Ca in all three materials, whilst other elements such as Al, Si and S were also found., Conclusions: MTA-Ang, MTA-HP and NeoMTA-P were associated with biological effects on hDPSCs in terms of cell proliferation, morphology, migration and attachment., (© 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Aims: To investigate the cytotoxicity and bioactivity of several pulpotomy materials: Biodentine (Septodont, Saint-Maur-des-Fosses, France) MTA (Angelus, Londrina, PR, Brazil), Theracal LC (Bisco Inc., Schamburg, IL, USA) and IRM (Dentsply DeTrey GmbH, Konstanz, Germany), after contact with stem cells isolated from human exfoliated primary teeth (SHEDs)., Methodology: SHEDs were cultured in the presence of the eluates of various pulpotomy materials for 24, 48 and 72 h. Cell viability was determined by mitochondrial dehydrogenase enzymatic (MTT) assay. Apoptosis and changes in cell phenotype were evaluated by flow cytometry. Also, an in vitro scratch wound-healing assay was used to determine their effects on cell migration. To assess cell morphology and attachment to the different pulpotomy materials, SHEDs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). Finally, the deposition of a calcified matrix in presence of these materials was verified by Alizarin Red staining. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α = 0.05)., Results: Cell viability in the presence of Biodentine eluates was significantly higher to that obtained using complete medium alone (control; P < 0.01) and was also significantly higher than using MTA Angelus from 48 h of incubation (P < 0.01). However, Theracal LC and IRM were associated with low rates of cell viability (P < 0.001). Similar results were obtained in an apoptosis assay. In addition, SHEDs maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of Biodentine. SEM studies revealed a suitable proliferation rate, cell spreading and attachment, especially when using Biodentine and MTA Angelus discs. Finally, Biodentine eluates significantly induced calcified matrix deposition from 7 days of culture (P < 0.01)., Conclusions: Biodentine exhibited better cytocompatibility and bioactivity than MTA Angelus, Theracal LC and IRM., (© 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Aim: To evaluate the biocompatibility of three calcium silicate-based endodontic sealers, Bioroot BC Sealer (Septodont, Saint-Maur-des-Fosses, France), Endoseal MTA (EndoSeal, Maruchi, Seoul, Korea) and Nano-ceramic Sealer (B&L Biotech, Fairfax, VA, USA) (NCS), on human periodontal ligament stem cells (hPDLSCs)., Methodology: Human periodontal ligament stem cells were cultured in the presence of various endodontic sealer eluates for 24 h. Cell viability was determined using the MTT assay. Cell death and changes in phenotype induced by the set endodontic sealer eluates were evaluated through flow cytometry. Also, an in vitro scratch wound-healing model was used to determine their effects in cell migration. Finally, to assess cell morphology and attachment to the different sealers, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). One-way analysis of variance (anova) followed by a Bonferroni post-test was performed (P < 0.05)., Results: At 24 h, cell spreading was evident in the presence of Bioroot BC Sealer (BR) and Nano-ceramic Sealer (NCS), but not Endoseal MTA (ES). At 72 h, BR and NCS exhibited high and moderate cell proliferation, respectively, whereas ES revealed low rates of cell proliferation (P < 0.05). Similar results were obtained in a cell death assay. In addition, hPDLSCs maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of BR. Finally, SEM studies revealed a good degree of proliferation, cell spreading and attachment, especially when using BR and NCS discs., Conclusions: BR and NCS were associated with better cytocompatibility than ES. Further in vitro and in vivo investigations are required to confirm the suitability of these calcium silicate-based endodontic sealers for clinical application., (© 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Objective: To evaluate the cytoprotective effects of melatonin (MLT) on zoledronic acid (ZA)-treated human osteoblasts., Methods: Human osteoblasts were exposed to ZA (1, 5, 10, 50, 100 and 300 μM) and MLT (1, 10, 50, 100 y 200 μM) for 24, 48 and 72 h of incubation, to evaluate their effects on cell viability., Results: As ZA concentration increased, greater reductions in cell viability of human osteoblasts were induced whether at 24, 48 or 72 h incubation. At 24 h incubation with MLT, greatest cell viability was obtained when low dose of MLT was applied (without significant differences); 48 and 72 h incubation presented the greatest cell viability with the highest MLT concentrations (100 and 200 μM). MLT at concentrations of 100 and 200 μM would appear to have a certain cytoprotective effect on ZA-treated human osteoblasts with low concentrations of ZA (1 y 5 μM), whether at 24, 48 or 72 h; however, at ZA concentrations ≥10 μM the possible cytoprotective effects of MLT were low at 24 h incubation., Conclusions: MLT has a cytoprotective effect on ZA-treated human osteoblasts and could represent a promising preventative alternative for patients at risk of bisphosphonate-related osteonecrosis of the jaw., (Copyright © 2017 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.)

Background: Leukaemia represents 30-40% of all paediatric malignant tumours and is the main cause of death in patients aged <15 years. One of the main complications in these patients is infection, which may often occur in the oral cavity. Chemotherapy-related oral health problems may be reduced by oral healthcare strategies based on the International Caries Detection and Assessment System (ICDAS) and Caries Management by Risk Assessment (CABRA)., Case Report: A case is reported of a 14-year-old girl treated for leukaemia who presented with established dental caries lesions which were classified and treated according to ICDAS and CABRA protocols. After three, no new caries was observed., Follow-Up and Conclusion: ICDAS and CAMBRA provide useful and effective guidance for the avoidance of dental and systemic problems. Their introduction into standard practice could reduce the legal difficulties derived from dental treatment in these patients.

Introduction: The aim of the present study was to evaluate the in vitro cytotoxicity of endodontic sealers (GuttaFlow Bioseal, GuttaFlow2, and MTA Fillapex) on human periodontal ligament stem cells (hPDLSCs). As a reference, AH Plus was compared with the more recent endodontic sealers regarding cell viability and cell attachment., Methods: Biological testing was carried out in vitro on hPDLSCs. Cell viability assay was performed by using eluates from each endodontic sealer. To assess cell morphology and attachment to the different sealers, the hPDLSCs were directly seeded onto the material surfaces and analyzed by scanning electron microscopy. Chemical composition of the sealers was determined by energy-dispersive x-ray, and eluates were analyzed by inductively coupled plasma mass spectrometry. Statistical differences were assessed by analysis of variance and Tukey test (P < .05)., Results: Cell viability was evident after 24 hours in the presence of GuttaFlow Bioseal and GuttaFlow 2 but not in the case of AH Plus or MTA Fillapex. At 168 hours, GuttaFlow Bioseal and GuttaFlow 2 exhibited high and moderate cell viability, respectively, whereas AH Plus and MTA Fillapex revealed low rates of cell cell viability (P < .001). Finally, scanning electron microscopy studies revealed a high degree of proliferation, cell spreading, and attachment, especially when using GuttaFlow Bioseal disks., Conclusions: GuttaFlow Bioseal and GuttaFlow2 showed lower cytotoxicity than MTA Fillapex and AH plus. Further in vitro and in vivo investigations are required to confirm the suitability of GuttaFlow Bioseal for clinical application., (Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Background: Recent times have witnessed a significant increase in the number of patients affected by problems related to oncological treatment Aims of this study is to evaluate dental affectation among patients awaiting hematopoietic progenitor cell transplant (HPCT), and they showed high caries risk, so it should establish a protocol prior to transplantation., Material and Methods: The study included 72 patients due for HPCT. Clinical and radiological explorations were performed and oral photos taken. The amount of caries, missing teeth and fillings were registered for each patient. CAO, DMFS and Restoration Indices were calculated., Results: 83% of patients presented caries. 48 patients (67%) had lost at least one tooth. Only 32 patients (44%) had received some sort of conservative treatment. The average CAO index value obtained was 10.37. The DMFS index showed an average of 27.06 affected surfaces. Of the 72 patients studied, 40 (56%) showed a restoration index value of zero., Conclusions: These patients presented a high number of carious teeth and a low restoration index. The presence of so many possible septic foci in an individual, who will later become susceptible to infection, highlights the importance of preventative treatment and bucco-dental restoration within this patient population. These patients with a high caries risk can be treated with CAMBRA system. Key words: Hematopoietic progenitor cell transplantation, high caries risk, state of oral health, haematological disease, CAMBRA system., Competing Interests: Conflict of interest statement:To ensure balance, independence, objectivity and scientific rigor each authors disclose any conflict of interest that may have a direct bearing on the subject matter of the paper.

Alpha (α)-thalassemias are the most common genetic disorder of hemoglobin (Hb) synthesis, affecting up to 5% of the world's population. These congenital hemolytic anemias induce extramedullary hematopoiesis, including the liver, spleen, sinuses, and the diploic spaces of the skull. Oral health problems in patients with thalassemias are mostly related to a varied degree of facial deformities, malocclusions, and/or dental arch dimensions. We present a case with a 49-year-old man, diagnosed with homozygous α thalassemia that came to the Faculty of Dentistry at the University of Murcia for a dental treatment. It was observed that the patient had an unusual mandibular manifestation of hematopoiesis., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Aim: To investigate in vitro the cytocompatibility of the calcium silicate-containing endodontic sealers MTA Fillapex and TotalFill BC Sealer on human periodontal ligament stem cells (hPDLSCs) by assaying their biological responses and compare them with that observed when using an epoxy resin-based sealer (AH Plus)., Methodology: Specimens from the three different endodontic sealers were eluated with culture medium for 24 h. The cytotoxicity of these eluates was evaluated using the MTT assay. In addition, an in vitro scratch wound healing model was used to determine their effects on cell migration. Cell adhesion to collagen type I after treatment with the different sealer eluates was also measured, whereas cytotoxicity was determined using the DNA-specific fluorochrome Hoechst 33342. Finally, to assess cell morphology and attachment to the different sealers, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). One-way analysis of variance (anova) followed by a Bonferroni post-test were performed (P < 0.05)., Results: hPDLSCs exposed to different dilutions of TotalFill BC Sealer eluates had significantly higher cell proliferation compared with that observed when cells were treated with AH Plus and MTA Fillapex eluates (P < 0.001). In addition, TotalFill eluates were associated with significantly increased cell adhesion to collagen type I and migration of hPDLSCs in a concentration-dependent manner than displayed after treatment with MTA Fillapex or AH Plus eluates (P < 0.001). Moreover, TotalFill BC Sealer-induced cytotoxicity was significantly lower than observed using AH Plus and MTA Fillapex eluates (P < 0.001). Finally, SEM studies revealed suitable proliferation, cell spreading and attachment, especially when using TotalFill BC Sealer discs., Conclusion: TotalFill BC Sealer exhibited a higher cytocompatibility than AH Plus and MTA Fillapex. Further investigations using in vivo animal models are required to validate the potential biological responses of TotalFill BC Sealer on hPDLSCs., (© 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Graphene represents one of the most interesting additions to the tissue engineering toolbox. Novel graphene-based composites are required to improve the beneficial graphene properties in terms of tridimensional polymeric structure, conferring a higher mechanical strength and favoring the differentiation of human mesenchymal stem cells. Here, we have demonstrated in a wide range of composite combinations, the successful use of graphene and silk-fibroin constructs for future bioengineering applications in the field of clinical regenerative dentistry using human periodontal ligament stem cells. Our results provide exciting new data for the development of suitable scaffolds that allow good cell engrafting, preservation of cell viability and proliferation, promotion of spontaneous osteoblastic differentiation, and importantly, stimulation of a higher cementum physiological synthesis than using other different available biomaterials.

Background: The clinical management of medication-related osteonecrosis of the jaw (MRONJ) in patients treated with bisphosphonates and other antiresorptive agents is subject to controversy. The American Association of Oral and Maxillofacial Surgeons (AAOMS) has developed guidelines for the correct management of the disorder which are revised and updated by a panel of experts., Material and Methods: The present systematic review analyzes the different treatments currently used to treat this clinical condition, based on the PRISMA® (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) statement published in 2009. An electronic Medline search was made of the PubMed database, covering the period 2006-2014. The last search date was 31 December 2014., Results: A total of 29 articles were selected from the initial search according to the different drugs implicated in the appearance of osteonecrosis; the treatment modality used according to the stage of the disease; and the recorded success rate., Conclusions: It is currently still recommended that the management of MRONJ should be decided according to the stage of the disease - conservative treatment being preferred in early stages without symptoms, while surgical management is preferred in the case of bone exposure with symptoms., Competing Interests: The authors have declared that no conflict of interest exist.

Objective: Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a common clinical complication in patients receiving bisphosphonate therapy. Furthermore, melatonin has been proposed as a therapeutic drug for the oral cavity due to its antioxidant properties. This study aimed to evaluate the cytoprotective effects of melatonin on zoledronic acid (ZA)-treated human mesenchymal stem cells from periodontal ligament (PDLSCs) and bone marrow (BMMSCs)., Methods: PDLSCs and BMMSCs were exposed to ZA, melatonin or ZA + melatonin for 72 h. Cell proliferation was measured by a colorimetric assay, whereas their mesenchymal phenotype was analyzed by flow cytometry., Results: Proliferation assays showed that BMMSCs presented higher ZA resistance than PDLSCs, as well as a difference in response to the simultaneous treatment of ZA + melatonin. Using PDLSCs, high doses of melatonin significantly increased their proliferation, whereas lower concentrations were enough to enhance ZA-treated BMMSC proliferation. Moreover, PDLSCs displayed a CD90/CD105 downregulation and CD73 upregulation in response to ZA, which was more pronounced in response to melatonin. Furthermore, ZA or ZA + low doses of melatonin induced a decrease of expression of CD90/CD105/CD73 on BMMSCs, while a higher concentration recovered CD73 levels., Conclusion: These results suggest that melatonin has a cytoprotective effect on ZA-treated PDLSCs and BMMSCs. Thus, it could be used for BRONJ prevention., (Copyright © 2015 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.)

Aims: To establish whether or not the state of patient oral health can influence the occurrence and/or severity of oral mucositis during hematopoietic progenitor cell transplantation (HPCT)., Materials and Methods: The study included 72 patients awaiting HPCT. Prior to transplantation, clinical exploration and radiology were carried out and oral photographs were taken. This evaluated the extent of caries present, the number of missing teeth and the number of dental fillings in each patient; CAO (Caries and Obturations Index) DMFS (Decayed, Missing, and Filled Surfaces) and Restoration Indices were calculated. Gingival pathology was also examined by means of the Ainamo and Bay Gingival Bleeding Index. O'Leary's Plaque Index was used to evaluate the level of patient oral hygiene. This data was analyzed to see if it exercised any influence on the mucositis grade suffered during HPCT., Results: 96,87% of patients suffered some degree of mucositis during their treatment by the Transplant Unit. The grade of mucositis was seen to be influenced by the number of missing teeth (ANOVA p<0.016) and by the DMFS Index (ANOVA p< 0.038). Although this was not one of the aims of this study, patient age and the administration of colony-stimulating factors were also seen to influence these clinical manifestations., Conclusions: The state of prior oral health can influence decisively the mucositis suffered during transplantation.

Cerebrovascular accident (CVA) is the third cause of death in industrialized countries, following cardiovascular disease and cancer. It is therefore a significant public health issue, not only due to its high incidence, but also to the high costs involved in the physical and psychological rehabilitation of these patients. Dental Practitioners, as health care providers, ought to play their part in this issue and contribute, within their means, to the early detection of patients at risk of having a CVA. Since the eighties, different authors have described the possibility of detecting calcified atheroma plaques located at carotid artery bifurcation through panoramic radiograph. In this way, the Dental Practitioner s possibilities in this field have been extended. However, this new use of panoramic radiograph must overcome certain obstacles before it is implemented as a new screening method for patients at risk of having a CVA. Amongst these, we would have, on the one hand, the assessment of the real clinical significance, as regards prognosis, of atheroma plaque calcification as well as its usefulness as a factor for predicting the appearance of CVA symptoms and, on the other hand, the possibility of making a correct differential diagnosis regarding other calcified structures that may appear on panoramic radiograph.

Oral mucositis is the inflammation that takes place in the oral epithelium, as a result of antineoplastic treatments such as radiotherapy, chemotherapy or bone marrow transplant, being very frequent in these treatments for oncohematologic disease. The consequences of this inflammation, not only affect the quality of life of the patient, but can also suppose a limitation in the application of the treatment, as well as an increase in the hospital stay and therapeutic costs. A main obstacle for the study of the mucositis, has been the lack of a system adapted for its valuation by means of the oral examination. Methods developed to measure and quantify the changes produced in oral epithelium as a result of treatment of cancer can be very varied from more simple methods, such as general scales with four or five degrees of severity that link the mucositis to the state of oral health, to specific scales of treatment. In this last type of scale the type of antineoplastic treatment that gave rise to the mucositis is identified giving a global severity score for the mucositis. The establishment of a common scale for the evaluation of mucositis is important, not only for clinical purposes but also for the investigation of the degree of toxicity of the different therapeutic regimes that give rise to the mucositis.

Childhood neoplasias have become increasingly important in recent years in the ambit of paediatric medicine. This phenomenon has been accompanied by a spectacular improvement in the treatment of childhood cancer, long-term survival rates reaching 90% in the case of some tumours. A corollary of this success is the obligation to provide new and improved medical assistance both as regards the possible prevention of any alterations and, if possible, the avoidance of complications derived from the neoplasm itself and its treatment. Among possible secondary effects are oral manifestations of a chronic or acute nature, which may cause great discomfort, act as foci of systemic infections or have long-term after effects, all of which will depend on the exact moment of the child s development that treatment is undertaken. The incidence and severity of most oral complications is associated with pre-existent factors, such as caries, gingivitis or generally poor hygiene, which strongly affect the beginning, increase and persistence of the same. It is to be decried that a problem in the buccal cavity is allowed to develop, which a simple preventative measure, simple hygiene or dental conservation treatment could prevent or reduce.