Adhesive Performance of Pit and Fissure Sealants on Deproteinized Enamel with Different Proteolytic Agents: In Vitro Study.

The objective of this work was to assess the efficacy of different proteolytic agents on the bond strength of pit and fissure sealants to bovine enamel. Eighty-four bovine enamel specimens were randomly assigned in groups according to the pit and fissure sealant applied (HelioSeal F or Dyad Flow). Then, the specimens were subdivided according to the proteolytic agent used (n = 7): Group 1, distilled water (control); Group 2, 10 wt.% Tergazyme®; Group 3, 10 wt.% ZYME®; Group 4, 10% papain gel; Group 5, 10% bromelain gel; and Group 6, 5.25 wt.% sodium hypochlorite. The cell viability of the proteolytic solutions was assessed through the MTT assay. The proteolytic agents were applied on the enamel surface prior to the acid-etching procedure; then, the pit and fissure sealants were placed. The micro-shear bond strength was evaluated after 24 h or 6 months of water storing at 37 °C. Representative SEM images were taken for each experimental group. The bond strength data were statistically analyzed by a three-way ANOVA test using a significance level of α = 0.05. Bromelain and papain proteolytic solutions did not exert any cytotoxic effect on the human dental pulp cells. After 24 h and 6 months of aging, for both pit and fissure sealants, sodium hypochlorite, papain, bromelain, and Tergazyme® achieved statistically significant higher bond strength values (p < 0.05). Irrespective of the deproteinizing agent used, Dyad Flow resulted in a better bond strength after 6 months of aging. The type 1 etching pattern was identified for sodium hypochlorite, papain, and bromelain. Tergazyme®, papain, and bromelain demonstrated efficacy in deproteinizing enamel surfaces prior to acid etching, leading to the improved bond strength of pit and fissure sealants. Clinically, this suggests that these proteolytic agents can be considered viable alternatives to traditional methods for enhancing sealant retention and longevity. Utilizing these agents in dental practice could potentially reduce sealant failures.

Gelatin-Chitosan Hydrogel Biological, Antimicrobial and Mechanical Properties for Dental Applications.

Chitosan, a natural polysaccharide sourced from crustaceans and insects, is often used with hydrogels in wound care. Evaluating its cytotoxicity and antimicrobial properties is crucial for its potential use in dentistry. OBJECTIVE: To investigate the mechanical properties of gelatin hydrogels based on decaethylated chitosan and antimicrobial activity against Streptococcus mutans and their biological effects with stem cells from apical papilla (SCAPs). MATERIAL AND METHODS: Gelatin-chitosan hydrogels were synthesized at concentrations of 0%, 0.2% and 0.5%. Enzymatic and hydrolytic degradation, along with swelling capacity, was assessed. Fourier transform infrared spectroscopy (FTIR) analysis was employed to characterize the hydrogels. The interaction between hydrogels and SCAPs was examined through initial adhesion and cell proliferation at 24 and 48 h, using the Thiazolyl Blue Tetrazolium Bromide (MTT assay). The antimicrobial effect was evaluated using agar diffusion and a microdilution test against S. mutans. Uniaxial tensile strength (UTS) was also measured to assess the mechanical properties of the hydrogels. RESULTS: The hydrogels underwent hydrolytic and enzymatic degradation at 30, 220, 300 min and 15, 25, 30 min, respectively. Significantly, (p < 0.01) swelling capacity occurred at 20, 40, 30 min, respectively. Gelatin-chitosan hydrogels' functional groups were confirmed using vibrational pattern analysis. SCAPs proliferation corresponded to 24 h = 73 ± 2%, 82 ± 2%, 61 ± 6% and 48 h = 83 ± 11%, 86 ± 2%, 44 ± 2%, respectively. The bacterial survival of hydrogel interaction was found to be 96 ± 1%, 17 ± 1.5% (p < 0.01) and 1 ± 0.5% (p < 0.01), respectively. UTS showed enhanced (p < 0.05) mechanical properties with chitosan presence. CONCLUSION: Gelatin-chitosan hydrogels displayed favorable degradation, swelling capacity, mild dose-dependent cytotoxicity, significant proliferation with stem cells from apical papilla (SCAPs), substantial antimicrobial effects against S. mutans and enhanced mechanical properties. These findings highlight their potential applications as postoperative care dressings.

Cytotoxicity, Differentiation, and Biocompatibility of Root-End Filling: A Comprehensive Study.

Assessing the biocompatibility of endodontic root-end filling materials through cell line responses is both essential and of utmost importance. This study aimed to the cytotoxicity of the type of cell death through apoptosis and autophagy, and odontoblast cell-like differentiation effects of MTA, zinc oxide-eugenol, and two experimental Portland cements modified with bismuth (Portland Bi) and barium (Portland Ba) on primary cell cultures. Material and methods: The cells corresponded to human periodontal ligament and gingival fibroblasts (HPLF, HGF), human pulp cells (HPC), and human squamous carcinoma cells from three different patients (HSC-2, -3, -4). The cements were inoculcated in different concentrations for cytotoxicity evaluation, DNA fragmentation in electrophoresis, apoptosis caspase activation, and autophagy antigen reaction, odontoblast-like cells were differentiated and tested for mineral deposition. The data were subject to a non-parametric test. Results: All cements caused a dose-dependent reduction in cell viability. Contact with zinc oxide-eugenol induced neither DNA fragmentation nor apoptotic caspase-3 activation and autophagy inhibitors (3-methyladenine, bafilomycin). Portland Bi accelerated significantly (p < 0.05) the differentiation of odontoblast-like cells. Within the limitation of this study, it was concluded that Portland cement with bismuth exhibits cytocompatibility and promotes odontoblast-like cell differentiation. This research contributes valuable insights into biocompatibility, suggesting its potential use in endodontic repair and biomimetic remineralization.

Decellularized Scaffolds of Nopal (Opuntia Ficus-indica) for Bioengineering in Regenerative Dentistry.

Opuntia Ficus-indica, or nopal, is traditionally used for its medicinal properties in Mexico. This study aims to decellularize and characterize nopal (Opuntia Ficus-indica) scaffolds, assess their degradation and the proliferation of hDPSC, and determine potential pro-inflammatory effects by assessing the expression of cyclooxygenase 1 and 2 (COX-1 and 2). The scaffolds were decellularized using a 0.5% sodium dodecyl sulfate (SDS) solution and confirmed by color, optical microscopy, and SEM. The degradation rates and mechanical properties of the scaffolds were determined by weight and solution absorbances using trypsin and PBS and tensile strength testing. Human dental pulp stem cells (hDPSCs) primary cells were used for scaffold-cell interaction and proliferation assays, as well as an MTT assay to determine proliferation. Proinflammatory protein expression of COX-I and -II was discovered by Western blot assay, and the cultures were induced into a pro-inflammatory state with interleukin 1-ß. The nopal scaffolds exhibited a porous structure with an average pore size of 252 ± 77 µm. The decellularized scaffolds showed a 57% reduction in weight loss during hydrolytic degradation and a 70% reduction during enzymatic degradation. There was no difference in tensile strengths between native and decellularized scaffolds (12.5 ± 1 and 11.8 ± 0.5 MPa). Furthermore, hDPSCs showed a significant increase in cell viability of 95% and 106% at 168 h for native and decellularized scaffolds, respectively. The combination of the scaffold and hDPSCs did not cause an increase in the expression of COX-1 and COX-2 proteins. However, when the combination was exposed to IL-1ß, there was an increase in the expression of COX-2. This study demonstrates the potential application of nopal scaffolds in tissue engineering and regenerative medicine or dentistry, owing to their structural characteristics, degradation properties, mechanical properties, ability to induce cell proliferation, and lack of enhancement of pro-inflammatory cytokines.

Natural Bioactive Epigallocatechin-Gallate Promote Bond Strength and Differentiation of Odontoblast-like Cells.

The (-)-Epigallocatechin-gallate (EGCG) metabolite is a natural polyphenol derived from green tea and is associated with antioxidant, biocompatible, and anti-inflammatory effects. OBJECTIVE: To evaluate the effects of EGCG to promote the odontoblast-like cells differentiated from human dental pulp stem cells (hDPSCs); the antimicrobial effects on Escherichia coli, Streptococcus mutans, and Staphylococcus aureus; and improve the adhesion on enamel and dentin by shear bond strength (SBS) and the adhesive remnant index (ARI). MATERIAL AND METHODS: hDSPCs were isolated from pulp tissue and immunologically characterized. EEGC dose-response viability was calculated by MTT assay. Odontoblast-like cells were differentiated from hDPSCs and tested for mineral deposition activity by alizarin red, Von Kossa, and collagen/vimentin staining. Antimicrobial assays were performed in the microdilution test. Demineralization of enamel and dentin in teeth was performed, and the adhesion was conducted by incorporating EGCG in an adhesive system and testing with SBS-ARI. The data were analyzed with normalized Shapiro-Wilks test and ANOVA post hoc Tukey test. RESULTS: The hDPSCs were positive to CD105, CD90, and vimentin and negative to CD34. EGCG (3.12 µg/mL) accelerated the differentiation of odontoblast-like cells. Streptococcus mutans exhibited the highest susceptibility < Staphylococcus aureus < Escherichia coli. EGCG increased (p < 0.05) the dentin adhesion, and cohesive failure was the most frequent. CONCLUSION: (-)-Epigallocatechin-gallate is nontoxic, promotes differentiation into odontoblast-like cells, possesses an antibacterial effect, and increases dentin adhesion.

Fracture Resistance of Endodontically Restored Teeth with Two System Root Post

ABSTRACT: The objective of this study was to determinate the fracture resistance of endodontically treated teeth and restored with two root post systems: i) resin post, ii) fiber post. A total of 60 teeth were freshly extracted, endodontic ally treated and randomly divided in two groups (n= 30/each group) for standardized restoration; Group 1 (Group R): Resin post and resin restoration, Group 2 (Group FP): Fiberglass post and resin restoration. Both groups' samples were mounted in a metallic base at 135º to allow them to be stabilized and held in the universal testing machine by applying a vertical force at cross speed of 1mm/min. Data were recorded in Newtons (N) Previous to test the fracture resistance; all samples were stored in distilled water at 37 ºC for 24 hours. Data were subject to the Saphiro-Wilk test for normality distribution and Student's t test. Significance was considered at 0.05 values. The values of fiber post group showed normal distribution compared to the resin group, demonstrating less variability among the values. The group FP displayed higher fracture resistance (299.77±100 N) than group R (205.57±86.40 N), with significant differences (p= 0.00002). The greatest fracture resistance was recorded for the group having fiber post reinforced and composite cores. It is suggested that fiberglass post restoration is the first option when endodontic treatment requires core restoration.

RESUMEN: El objetivo del estudio fue determinar la resistencia a la fractura de dientes tratados endodónticamente y restaurados con dos sistemas de endopostes radiculares: I) poste de resina, II) poste de fibra. Un total de 60 dientes recién extraídos fueron tratados endodónticamente y divididos al azar en dos grupos (n= 30/ cada grupo) para la restauración estandarizada; Grupo 1 (Grupo R): Pilares de resina y restauración, Grupo 2 (Grupo FP): Pilares de fibra de vidrio y restauración de resina. Las muestras de ambos grupos se montaron en una base metálica a 135º para permitir su estabilización y sujeción en la máquina universal de ensayos aplicando una fuerza vertical a velocidad transversal de 1 mm/min. Los datos se registraron en Newtons (N), para probar la resistencia a la fractura; todas las muestras se almacenaron en agua destilada a 37 ºC durante 24 horas. Los datos se sometieron a la prueba de normalidad de Saphiro-Wilk y la pruebas t de Student. La significancia se consideró con un valor de 0,05. Los valores del grupo de postes de fibra mostraron una distribución normal en comparación con el grupo de resinas, demostrando menor variabilidad entre los valores. El grupo FP mostró mayor resistencia a la fractura (299,77±100 N) que el grupo R (205,57±86,40 N) que el grupo con diferencias significativas (p= 0,00002). La mayor resistencia a la fractura se registró para el grupo que tenía núcleos compuestos y reforzados con postes de fibra. Se sugiere que la restauración posterior de fibra de vidrio es la primera opción cuando el tratamiento de endodoncia requiere una restauración del núcleo.

CRISPR/Cas gene-editing technology and its advances in dentistry.

A recent discovery of revolutionary Clustered regularly interspaced palindromic repeats (CRISPR) is a gene-editing tool that provides a type of adaptive immunity in prokaryotic organisms, which is currently used as a revolutionizing tool in biomedical research. It has a mechanism of correcting genome errors, turning on/off genes in cells and organisms. Most importantly playing a crucial function in bacterial defence by identifying and destroying Deoxyribonucleic acid (DNA) segments during bacteriophage invasions since the CRISPR-associated protein 9 (Cas9) enzyme recognizes and cleaves invasive DNA sequences complementary to CRISPR. Therefore, researchers employ this biological device to manipulate the genes to develop new therapies to combat systemic diseases. Currently, the most significant advance at the laboratory level is the generation of cell and animal models, functional genomic screens, live images of the cell genome, and defective DNA repairs to find the cure for genetic disorders. Even though this technology has enormous biomedical applications in various sectors, this review will summarize CRISPR/Cas emphasizing both the therapeutic and diagnostic mechanisms developed in the field of dentistry and the promising attempts to transfer this technology to clinical application. Finally, future developments are also described, which proposes to use CRISPR/Cas systems for prospective clinical dentistry applications.

[Cancer stem cells in oral squamous cell carcinoma. Literature review]. / Células troncales cancerígenas en carcinoma oral de células escamosas. Revisión de la literatura.

Objective: To perform a literature review on oral squamous cell carcinoma, the presence of cancer stem cells; their association with the course of the disease and therapeutic applications. Methods: : A search was performed in the PubMed database by entering the following algorithm: ((((neoplastic stem cells [MeSH Terms ]) OR (Cancer stem cells [Text Word ])) AND (Squamous Cell Carcinoma of Head and Neck [MeSH Terms])) AND (Oral squamous cell carcinoma [Text Word ]), to find articles in english published between 2012 and 2022. The PRISMA diagram was used to identify and select the articles. Results: A result of 49 articles was obtained; of which 27 were chosen according to the title and abstract in their association with the topic. In addition, 8 additional articles suggested by their relationship with the information previously searched were included. In total, 35 articles were evaluated. There has been found that tumoral cells in squamous oral carcinoma are heterogeneous since they include cancer stem cells wich possess characteristics of stem and neoplasic cells; which possess characteristics of stem cells as well as neoplastic cells; they have been associated with disease progression, recurrence, and metastasis and have been considered to be a key mechanism of therapy failure. Conclusions: The expression of stem cell markers in oral squamous cell carcinomas has been demonstrated and has contributed to their identification in oral squamous cell carcinomas and has been implicated in the behavior of cancer cells. New therapeutic measures aimed at eliminating cancer stem cells have been proposed and developed.

2D Nanosheets-A New Class of Therapeutic Formulations against Cancer.

Researchers in cancer nanomedicine are exploring a revolutionary multifaceted carrier for treatment and diagnosis, resulting in the proposal of various drug cargos or "magic bullets" in this past decade. Even though different nano-based complexes are registered for clinical trials, very few products enter the final stages each year because of various issues. This prevents the formulations from entering the market and being accessible to patients. In the search for novel materials, the exploitation of 2D nanosheets, including but not limited to the highly acclaimed graphene, has created extensive interest for biomedical applications. A unique set of properties often characterize 2D materials, including semiconductivity, high surface area, and their chemical nature, which allow simple decoration and functionalization procedures, structures with high stability and targeting properties, vectors for controlled and sustained release of drugs, and materials for thermal-based therapies. This review discusses the challenges and opportunities of recently discovered 2D nanosheets for cancer therapeutics, with special attention paid to the most promising design technologies and their potential for clinical translation in the future.

Cytotoxic and anti-inflammatory effects of chitosan and hemostatic gelatin in oral cell culture.

Chitosan is a biopolymer with bactericidal/bacteriostatic effect, biocompatible and biodegradable. It has been used in tissue engineering to replace tissues partially or completely by releasing bioactive materials or influencing cell growth, usually in regenerative medicine and dentistry. The aim of this study was to evaluate the cytotoxic and anti-inflammatory effect of chitosan alone or with hemostatic gelatin (Spongostand®) in cultures of human pulp cells (HPC), human gingival fibroblasts (HGF) and mouse pre-osteoblasts (MC3T3-E1, ATCC). HPC and HGF were isolated from patients. Cells were subcultured in DMEM. Chitosan was inoculated at different concentrations (0-0.5%) and hemostatic gelatins impregnated with chitosan (0.19%) were placed directly in the presence of cells and incubated for 24 hours. Cell viability was determined by MTT method and mean cytotoxic concentration (CC50) was calculated from the dose-response curve. Anti-inflammatory effect was calculated from the in vitro gingivitis model induced with interleukin 1beta (IL-1ß) in HGF and protein detection. The data were subjected to Shapiro-Wilk, Kruskal-Wallis and Mann-Whitney tests. Experiments were performed in triplicate of three independent assays. Cell viability of HPC, HGF and MC3T3-E1 in contact with chitosan decreased significantly (p<0.05). The HPC were the most sensitive (CC50= 0.18%), followed by HGF (CC50= 0.18%) and MC3T3-E1 (CC50= 0.19%). The cytotoxicity of gelatins impregnated with chitosan decreased cell viability of HGF and HPC by 11% and 5%, respectively. The proinflammatory effect was reduced significantly in the gingivitis model. To conclude, chitosan induces moderate cytotoxic effects alone or with hemostatic gelatin at 0.19%, in dose-dependent manner, with anti-inflammatory effects on human gingival fibroblasts. The use of chitosan as a biomaterial can be an excellent choice for use in regenerative dentistry.

El quitosano es un biopolímero con efecto bactericida/bacteriostático, biocompatible y biodegradable. Se ha utilizado en ingeniería de tejidos con el fin de reemplazar parcial o completamente los tejidos como material bioactivo o influyendo en el crecimiento celular, comúnmente, para medicina y odontología regenerativa. Evaluar el efecto citotóxico y antiinflamatorio del quitosano solo o con gelatina hemostática (Spongostand®) en cultivos con células pulpares humanas (HPC), fibroblastos gingivales humanos (HGF) y preosteoblastos de ratón (MC3T3-E1, ATCC). HPC, HGF se aislaron de pacientes. Las células se subcultivaron en DMEM. Se inoculó quitosano a diferentes concentraciones (0-0,5%) y se colocaron gelatinas hemostáticas impregnadas con quitosano (0,19%) directamente en presencia de células y se incubaron durante 24 horas. La viabilidad celular se determinó mediante el método MTT y se calculó la concentración citotóxica media (CC50) a partir de la curva dosis-respuesta. El efecto antiinflamatorio se calculó a partir del modelo de gingivitis in vitro inducido con interleucina 1ß (IL-1ß) en HGF. Los datos se sometieron a las pruebas de Shapiro-Wilk, Kruskal-Wallis y Mann-Whitney. Los experimentos se realizaron por triplicado de tres ensayos independientes. La viabilidad celular de HPC, HGF y MC3T3-E1 en contacto con el quitosano disminuyó significativamente la viabilidad celular (p<0.05). Las HPC fueron las más sensibles (CC50= 0,18%) seguido de HGF (CC50= 0,18%) y MC3T3-E1 (CC50= 0,19%). Las gelatinas impregnadas con quitosano mostraron una disminución en la viabilidad celular para HGF, HPC de 11% y 5% respectivamente y se redujo significativamente el efecto pro-inflamatorio en el modelo de gingivitis humano. El quitosano induce efectos citotóxicos moderados solo o con gelatina hemostática a 0,19% de forma dosis-dependiente con efectos antiinflamatorios en fibroblastos gingivales humanos. El uso de quitosano como biomaterial puede ser una excelente opción para su uso en odontología regenerativa.

Cytotoxic and Pro-Inflammatory Effects of Miniscrews Subject to Corrosion in Oral Cell Cultures.

Cytotoxic and pro-inflammatory effects of mini-screws subject to corrosion in oral cells culture. To analysis the products of corrosion of three different commercial orthodontic mini-screws and evaluate the cytotoxicity and pro-inflammatory effects in culture with human gingival fibroblast cells (HGFs) and human osteoblast-like bone surface cells (HBCs). An experimental in vitro study was carried out with 3 different type of mini-screws: Vector-Tas®, Forestadent ORTHOEasy®, Bio-Ray® (n = 30/gp). The samples were subjected to accelerated corrosion for 24, 48, 72 and 96 hours, which were observed with a stereomicroscope and scanning electron microscope. The corrosion products were analyzed by inductively coupled plasma with mass spectrometry. The direct and indirect cytotoxicity was tested in culture with HGFs and HBCs, cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The pro-inflammatory effect was determined by the expression of prostaglandin E2 (PGE2) with the ELISA test. The data were subjected to Shapiro-Wilks normality tests, paired t-tests and Tukey's post hoc ANOVA. The mini-screw topography showed significant morphological changes after corrosion. The main ions after corrosion were Al, Ti, and Fe. Corrosion products by direct and indirect contact with cells slightly reduced (P < 0.05) cell viability, considered non-cytotoxic. The expression of PGE2 was not increased by the presence of the corrosion products even in a previous pro-inflammatory state. The corrosion products were not cytotoxic and did not induce a pro-inflammatory state in culture with HGFs and HBCs.

Silver Nanoparticles from Annona muricata Peel and Leaf Extracts as a Potential Potent, Biocompatible and Low Cost Antitumor Tool.

Cancer is one of the most prevalent diseases in the world and requires new therapies for its treatment. In this context, the biosynthesis of silver nanoparticles (AgNPs) has been developed to treat different types of tumors. The Annona muricata plant is known for having anticancer activity. Its main compounds present in the leaves, stems and skin, allowing for its use as reducing agents. In this manuscript, AgNPs with leaf extract (AgNPs-LE) and fruit peel extract (AgNPs-PE) of A. muricata were biosynthesized obtaining an average nanoparticle diameter sizes smaller than 50 nm, being 19.63 ± 3.7 nm and 16.56 ± 4.1 nm, and with a surface plasmonic resonance (SPR) at 447 and 448 nm, respectively. The lactone functional group present in the LE and PE extracts was identified by the FTIR technique. The behavior and antiproliferation activity of AgNPs-LE and AgNPs-PE were evaluated in breast, colon and melanoma cancer cell lines. Our results showed that Annona muricata fruit peel, which is a waste product, has an antitumor effect more potent than leaf extract. This difference is maintained with AgNPs where the destruction of cancer cells was, for the first time, achieved using concentrations that do not exceed 3 µg/mL with a better therapeutic index in the different tumor strains. In conclusion, we present a low-cost one-step experimental setup to generate AgNPs-PE whose in-vitro biocompatibility and powerful therapeutic effect make it a very attractive tool worth exploiting.

Cytotoxic and anti-inflammatory effects of chitosan and hemostatic gelatin in oral cell culture / Efectos citotóxicos y antiinflamatorios del quitosano y la gelatina hemostática en cultivo celular oral

ABSTRACT Chitosan is a biopolymer with bactericidal/bacteriostatic effect, biocompatible and biodegradable. It has been used in tissue engineering to replace tissues partially or completely by releasing bioactive materials or influencing cell growth, usually in regenerative medicine and dentistry. The aim of this study was to evaluate the cytotoxic and anti-inflammatory effect of chitosan alone or with hemostatic gelatin (Spongostand®) in cultures of human pulp cells (HPC), human gingival fibroblasts (HGF) and mouse pre-osteoblasts (MC3T3-E1, ATCC). HPC and HGF were isolated from patients. Cells were subcultured in DMEM. Chitosan was inoculated at different concentrations (0-0.5%) and hemostatic gelatins impregnated with chitosan (0.19%) were placed directly in the presence of cells and incubated for 24 hours. Cell viability was determined by MTT method and mean cytotoxic concentration (CC50) was calculated from the dose-response curve. Anti-inflammatory effect was calculated from the in vitro gingivitis model induced with interleukin 1beta (IL-1β) in HGF and protein detection. The data were subjected to Shapiro-Wilk, Kruskal-Wallis and Mann-Whitney tests. Experiments were performed in triplicate of three independent assays. Cell viability of HPC, HGF and MC3T3-E1 in contact with chitosan decreased significantly (p<0.05). The HPC were the most sensitive (CC50= 0.18%), followed by HGF (CC50= 0.18%) and MC3T3-E1 (CC50= 0.19%). The cytotoxicity of gelatins impregnated with chitosan decreased cell viability of HGF and HPC by 11% and 5%, respectively. The proinflammatory effect was reduced significantly in the gingivitis model. To conclude, chitosan induces moderate cytotoxic effects alone or with hemostatic gelatin at 0.19%, in dose-dependent manner, with anti-inflammatory effects on human gingival fibroblasts. The use of chitosan as a biomaterial can be an excellent choice for use in regenerative dentistry.

RESUMEN El quitosano es un biopolímero con efecto bactericida/bacteriostático, biocompatible y biodegradable. Se ha utilizado en ingeniería de tejidos con el fin de reemplazar parcial o completamente los tejidos como material bioactivo o influyendo en el crecimiento celular, comúnmente, para medicina y odontología regenerativa. Evaluar el efecto citotóxico y antiinflamatorio del quitosano solo o con gelatina hemostática (Spongostand®) en cultivos con células pulpares humanas (HPC), fibroblastos gingivales humanos (HGF) y preosteoblastos de ratón (MC3T3-E1, ATCC). HPC, HGF se aislaron de pacientes. Las células se subcultivaron en DMEM. Se inoculó quitosano a diferentes concentraciones (0-0,5%) y se colocaron gelatinas hemostáticas impregnadas con quitosano (0,19%) directamente en presencia de células y se incubaron durante 24 horas. La viabilidad celular se determinó mediante el método MTT y se calculó la concentración citotóxica media (CC50) a partir de la curva dosis-respuesta. El efecto antiinflamatorio se calculó a partir del modelo de gingivitis in vitro inducido con interleucina 1β (IL-1β) en HGF. Los datos se sometieron a las pruebas de Shapiro-Wilk, Kruskal-Wallis y Mann-Whitney. Los experimentos se realizaron por triplicado de tres ensayos independientes. La viabilidad celular de HPC, HGF y MC3T3-E1 en contacto con el quitosano disminuyó significativamente la viabilidad celular (p <0.05). Las HPC fueron las más sensibles (CC50= 0,18%) seguido de HGF (CC50= 0,18%) y MC3T3-E1 (CC50= 0,19%). Las gelatinas impregnadas con quitosano mostraron una disminución en la viabilidad celular para HGF, HPC de 11% y 5% respectivamente y se redujo significativamente el efecto pro-inflamatorio en el modelo de gingivitis humano. El quitosano induce efectos citotóxicos moderados solo o con gelatina hemostática a 0,19% de forma dosis-dependiente con efectos antiinflamatorios en fibroblastos gingivales humanos. El uso de quitosano como biomaterial puede ser una excelente opción para su uso en odontología regenerativa.

Biological and physico-mechanical properties of poly (methyl methacrylate) enriched with graphene oxide as a potential biomaterial / Propiedades físicas-mecánicas-biológicas del poli (metilmetacrilato) enriquecido con óxido de grafeno como potencial biomaterial

Objective: To determine the cytotoxicity and effects of graphene oxide (GO) on cellular proliferation of gingival-fibroblasts, pulp-dental cells and human osteoblasts in culture, and to determine the physical, mechanical and biological properties of poly (methyl methacrylate) (PMMA) enriched with GO. Material and Methods: The GO was characterized with SEM. Cytotoxicity and cell proliferation were determined by the MTT bioassay. The physical mechanical tests (flexural strength and elastic modulus) were carried out with a universal testing machine. Sorption and solubility were determined by weighing before and after drying and immersion in water. Porosity was evaluated by visual inspection. Data were analyzed with Student's t-test and Tukey's posthoc ANOVA. Results: The GO has a heterogeneous morphology and a particle size of 66.67±64.76 µm. GO has a slight to no-cytotoxicity (>50-75% viability) at 1-30 days, and at 24 hours incubation of PMMA with GO significantly stimulates osteoblasts (45±8%, p<0.01). The physical and mechanical properties of PMMA with GO increase considerably without altering sorption, solubility and porosity. Conclusion: GO alone or with PMMA has an acceptable biocompatibility, could contribute to cell proliferation, cell regeneration and improve the physical mechanical properties of PMMA.

Objective: To determine the cytotoxicity and effects of graphene oxide (GO) on cellular proliferation of gingival-fibroblasts, pulpdental cells and human osteoblasts in culture, and to determine the physical, mechanical and biological properties of poly (methyl methacrylate) (PMMA) enriched with GO. Material and Methods: T he G O w as c haracterized with SEM. Cytotoxicity and cell proliferation were determined by the MTT bioassay. The physical-mechanical tests (flexural strength and elastic modulus) were carried out with a universal testing machine. Sorption and solubility were determined by weighing before and after drying and immersion in water. Porosity was evaluated by visual inspection. Data were analyzed with Student's t-test and Tukey's post-hoc ANOVA. Results: The GO has a heterogeneous morphology and a particle size of 66.67±64.76 ?m. GO has a slight to no-cytotoxicity (>50-75% viability) at 1-30 days, and at 24 hours incubation of PMMA with GO significantly stimulates osteoblasts (45±8%, p<0.01). The physical and mechanical properties of PMMA with GO increase considerably without altering sorption, solubility and porosity. Conclusion: GO alone or with PMMA has an acceptable biocompatibility, could contribute to cell proliferation, cell regeneration and improve the physical-mechanical properties of PMMA.

Antifungal biomaterial for reducing infections caused by Candida albicans in edentulous patients.

INTRODUCTION: Polymethylmethacrylate (PMMA) acrylic resins are used to make dentures for edentulous patients. OBJECTIVE: To find out the prevalence of Candida species in patients with and without removable prostheses from a dental clinic in León, Guanajuato, as well as to assess the antifungal effect and biological behavior of an experimental PMMA with silver nanoparticles for its possible application in prostheses. METHOD: To identify Candida species, smear samples were obtained from the palatal mucosa of 140 patients aged ≥ 60 years. The experimental PMMA with silver nnoparticles was placed in Candida albicans cultures, which were stained with the Live/Dead® kit for analysis under confocal microscopy; subsequently, it was implanted in Wistar rats in order to know its behavior in the surrounding tissues. RESULTS: Candida albicans was the most prevalent species in the evaluated patients, followed by Candida tropicalis and Candida krusei. The acrylic resin with silver nanoparticles significantly decreased the presence of Candida albicans. In the animal model, a discrete and controlled inflammatory reaction was found, which indicated biocompatibility of the acrylic resin that was used. CONCLUSIONS: It is possible for the nanostructured material with antifungal effect to be used in order to promote the reduction of oral Candida infections in edentulous patients.

INTRODUCCIÓN: Las resinas acrílicas de polimetilmetacrilato (PMMA) son utilizadas para elaborar dentaduras para pacientes edéntulos. OBJETIVO: Conocer la prevalencia de las especies de Candida en pacientes con y sin prótesis removibles de una clínica de odontología en León, Guanajuato; así como valorar el efecto antifúngico y el comportamiento biológico de un PMMA experimental con nanopartículas de plata para su posible aplicación en prótesis. MÉTODO: Para identificar las especies de Candida se obtuvieron muestras para frotis de la mucosa palatina de 140 pacientes con edad ≥ 60 años. El PMMA experimental con nanopartículas de plata fue colocado en cultivos de Candida albicans, los cuales fueron teñidos con el kit Live/Dead® para su análisis bajo microscopia confocal; posteriormente, se implantó en ratas Wistar para conocer su comportamiento en los tejidos circundantes. RESULTADOS: Candida albicans fue la especie más prevalente en los pacientes valorados, seguida de Candida tropicalis y Candida krusei. La resina acrílica con nanopartículas de plata disminuyó significativamente la presencia de Candida albicans. En el modelo animal se encontró reacción inflamatoria discreta y controlada, lo cual indicó la biocompatibilidad de la resina acrílica utilizada. CONCLUSIONES: Es posible utilizar el material nanoestructurado con efecto antifúngico para promover la reducción de infecciones orales por Candida en pacientes edéntulos.

Copper nanoparticles as nanofillers in an adhesive resin system: An in vitro study.

BACKGROUND: The incorporation of an antibacterial agent into an adhesive could improve its clinical performance. Some nanoparticles (NPs), including copper nanoparticles (Cu NPs), display an antibacterial effect. Therefore, Cu NPs could act as a nanofiller when added to an adhesive. OBJECTIVES: The aim of this study was to evaluate the antibacterial activity, cytotoxicity and shear bond strength (SBS) of an experimental dental adhesive with Cu NPs. MATERIAL AND METHODS: Different concentrations (0.0050 wt%, 0.0075 wt% and 0.0100 wt%) of Cu NPs were added to the adhesive. The distribution of Cu NPs in the polymer matrix was observed based on transmission electron microscope (TEM) images. The antimicrobial activity of the adhesive + Cu NPs was evaluated with the agar disk diffusion test against Staphylococcus aureus (S. aureus), Escherichia coli (E. coli) and Streptococcus mutans (S. mutans). The cytotoxicity assay was performed by means of the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method with human pulp cells (HPC). Additionally, the SBS tests were carried out (n = 31) and the modes of fracture were registered. The vestibular and lingual surfaces of each tooth were randomly assigned to the study groups (group I - control adhesive; group II - adhesive + 0.0100 wt% Cu NPs). The samples were statistically analyzed (p ≤ 0.05). RESULTS: The adhesive + 0.0100 wt% Cu NPs showed inhibition zones against the strains under study that were similar to, or slightly smaller than, the halos produced by chlorhexidine (CHX) and specific drugs for each strain (30 µg of cefotaxime against S. mutans and S. aureus, and 1.25/3.75 µg of sulfamethoxazole/ trimethoprim against E. coli). The control adhesive was moderately cytotoxic (relative cell viability of 36.7 ±0.8%), being more cytotoxic than Cu NPs themselves (58.3 ±0.1%). A significantly higher SBS was obtained for the adhesive + 0.0100 wt% Cu NPs (6.038 ±2.95 MPa) than for the control group (3.278 ±1.75 MPa). The modes of fracture in group I were almost equally distributed between adhesive and cohesive failures whereas in group II, the failure was mainly cohesive. CONCLUSIONS: The results of this study suggest that incorporating Cu NPs into an adhesive improves its SBS and provides it with antibacterial properties, without increasing its inherent cytotoxicity - 2 desirable characteristics for the dental adhesives of composites.

Antifungal and Cytotoxic Evaluation of Photochemically Synthesized Heparin-Coated Gold and Silver Nanoparticles.

Heparin-based silver nanoparticles (AgHep-NPs) and gold nanoparticles (AuHep-NPs) were produced by a photochemical method using silver nitrate and chloroauric acid as metal precursors and UV light at 254 nm. UV-Vis spectroscopy graphs showed absorption for AgHep-NPs and AuHep-NPs at 420 nm and 530 nm, respectively. TEM revealed a pseudospherical morphology and a small size, corresponding to 10-25 nm for AgHep-NPs and 1.5-7.5 nm for AuHep-NPs. Their antifungal activity against Candida albicans, Issatchenkia orientalis (Candida krusei), and Candida parapsilosis was assessed by the microdilution method. We show that AgHep-NPs were effective in decreasing fungus density, whereas AuHep-NPs were not. Additionally, the viability of human gingival fibroblasts was preserved by both nanoparticle types at a level above 80%, indicating a slight cytotoxicity. These results are potentially useful for applications of the described NPs mainly in dentistry and, to a lesser extent, in other biomedical areas.

Alargamiento de corona estético previo a rehabilitación protésica. Informe de caso / Aesthetic crown lengthening prior to prosthetic rehabilitation. Case report

RESUMEN: Introducción: Uno de los procedimientos realizados con mayor frecuencia para el restablecimiento de la estética en la zona anterosuperior es el alargamiento de corona, el cual está indicado ante la presencia de una longitud insuficiente de la corona clínica de un diente, cuando existe caries o fracturas subgingivales y para la mejora de la estética en pacientes con margen gingival desigual. Objetivo: El objetivo de este artículo es presentar el alargamiento de corona estético realizado en una paciente que presentaba márgenes discrepantes y pérdida de las papilas en la zona anterosuperior, la cual fue remitida del área de prótesis para poder realizar posterior rehabilitación protésica. Resultados: Mediante el procedimiento quirúrgico se logró la corrección y restablecimiento de los márgenes gingivales. Conclusiones: La zona anterosuperior es el área estéticamente más comprometida, por lo que es indispensable la planeación interdisciplinaria del tratamiento para asegurar el éxito del mismo.

ABSTRACT: Introduction: One of the most frequently performed procedures for aesthetics restoration in the maxillary anterior area is the crown lengthening, which is indicated in the presence of an insufficient length of the clinical crown of a tooth, of cavities or subgingival fractures and for the improvement of aesthetics in patients with uneven gingival margin. Objective: The aim of this article is to present the aesthetic crown lengthening performed in a patient with discrepant margins and loss of the papillae in the maxillary anterior area, referred from the prosthesis department for subsequent prosthetic rehabilitation. Results: Through the surgical procedure, the correction and restoration of the gingival margins were achieved. Conclusions: The maxillary anterior area is the most aesthetically compromised area, which is why the interdisciplinary planning of the treatment is essential to ensure its success.

Cytotoxicity of solutions recommended for the storage of avulsed teeth in cultures with periodontal ligament cells. / Citotoxicidad de soluciones recomendadas para el almacenamiento de dientes avulsionados en cultivo con células del ligamento periodontal.

Introduction: The medium for avulsed teeth storage until their reimplantation is key to the preservation of human periodontal ligament fibroblasts (HPLF). Objective: Our purpose was to compare the cytotoxic effect of milk and isotonic solution, used for the storage of avulsed teeth, on the preservation of HPLF. Method: A subculture of periodontal ligament fibroblasts was carried out with a density of 1:2 (3 ×105 cells/mL) and was incubated for 48 hours. The cells were divided in two groups, which were placed either in milk or isotonic solution for 24 hours at 5% CO2, 37 ºC and 95% humidity. The number of viable cells was determined with a colorimetric fast assay by the reduction of MTT and mitochondrial activity. Data were processed with the Shapiro-Wilk normality test, Student's t-test and paired Student's t-test (with significance set at 0.05). Results: The cells exposed to milk for 24 hours showed statistically significant cytotoxicity at concentrations of 0.09, 0.39, 0.78, 1.56, 3.125, 6.25 and 50%. HPLFs exposed to isotonic solution showed no significant reduction in the number of cells at concentrations of 25 and 50%. Conclusion: Isotonic solution appears to be better for HPLF 24-hour storage in comparison with whole milk.

Introducción: El medio de almacenamiento de los dientes avulsionados hasta su reimplante es vital para conservar los fibroblastos del ligamento periodontal humano (HPLF). Objetivo: Comparar el efecto citotóxico para conservar los HPLF de la leche y la solución isotónica para almacenamiento de dientes avulsionados. Método: Se realizó subcultivo de fibroblastos del ligamento periodontal con una densidad de 1:2 (3 × 105 células/mL), que fueron incubados por 48 horas. Se integraron dos grupos de células, que se colocaron en leche y solución isotónica durante 24 horas a 5 % de CO2, a 37°C y 95 % de humedad. El número de células viables fue determinado por colorimetría rápida por reducción de MTT y actividad mitocondrial. Los datos fueron sometidos a pruebas de normalidad de Shapiro-Wilk, t de Student y t de Student pareada (significación de 0.05). Resultados: Las células expuestas a la leche por 24 horas mostraron citotoxicidad estadísticamente significativa a concentraciones de 0.09, 0.39, 0.78, 1.56, 3.125, 6.25 y 50 %. Los HPLF expuestos a solución isotónica no mostraron reducción significativa del número de células a concentraciones de 25 y 50 %. Conclusión: La solución isotónica parece mejor para el almacenamiento de HPLF en 24 horas, comparada con la leche entera.

Tratamiento de la osteítis condensante en molar: informe de caso / Molar condensing osteitis treatment: Case report

En la actualidad, la principal causa por la que acuden los pacientes al odontólogo es el dolor dental, endonde la mayoría presenta un padecimiento pulpar o periapical irreversibles, que pueden estar asociados a factores traumáticos e irritativos. Sin embargo, pocosde ellos son asintomáticos, como la osteítis condensante que es escasamente mencionada en elámbito de la Endodoncia; por lo tanto, el objetivo de este caso clínico es el de proporcionar información acerca de la osteítis condensante siguiendo los lineamientos internacionales de Case Report (CARE). La osteítis condensante tiene una incidencia muy baja en pacientes y se debe diagnosticar correctamente al momento de tratar este tipo de lesiones con las diferentes herramientas de diagnóstico que se conocen. En este caso, se presenta un paciente del sexo femenino de 58 años de edad con un estado prediabético, que refiere un fractura del segundo molar inferior derecho, al cual radiográficamente se le encontróuna lesión periapical radiopaca en la raíz distal. Se muestra la secuencia del tratamiento, el manejo clínico y la rehabilitación.

At present, the main reason for patients to visit adentist is dental pain, where most of them presenta pulp or periapical irreversible condition, whichmay be associated with traumatic and irritative factors. However, few of them are asymptomatic as osteitiscondensing that is barely mentioned in thefield of endodontics. The aim of this case report isto provide information about the condensing osteitisfollowing international Case Report (CARE)guidelines. Condensing osteitis has a very low incidence in patients and should be correctly diagnosed with the different available diagnostic tools. In thiscase a 58-years-old female patient, with prediabeticstate, referred of a right lower second molar fracturewhich radiographically showed a radiopaque periapicallesion in the distal root of the molar. The sequence of treatment, clinical management and rehabilitation is presented.