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1.
Arch Oral Biol ; 131: 105273, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34600332

ABSTRACT

OBJECTIVES: The present study aimed to assess the oxidative stress and the viability of dental pulp cells stimulated by lipopolysaccharide (LPS) and submitted to photobiomodulation (PBM) with infrared light-emitting diode (LED, 850 nm). DESIGN: Three healthy primary teeth (n = 3) were collected and seeded in 24-well plates with 10 µg/mL of LPS to induce inflammatory mediator formation. The cells were irradiated (850 nm, 40 mW/cm2 and 80 mW/cm2) at the proposed radiant exposures of 0 (control), 4, 15, and 30 J/cm2 shortly after LPS supplementation. The tests were performed 24 h after irradiation to assess mitochondrial activity (MTT assay), the number of viable cells (Trypan Blue), cell morphology (Scanning Electron Microscopy - SEM), and the quantification of Nitric Oxide (NO) and Reactive Oxygen Species (ROS). The data were analyzed using Kruskal-Wallis and Dunn's tests (p < 0.05). RESULTS: The irradiated groups showed larger viable cells number than the non-irradiated group with LPS (p < 0.0001). All irradiation parameters decreased ROS concentrations after LPS application compared to the non-irradiated group (p < 0.05). All irradiation parameters enhanced the NO values compared to those of the control group (p < 0.05). The SEM images showed cells with regular morphology that adhered to the substrate. CONCLUSIONS: According to the parameters used in this study, the radiant exposure of 15 J/cm2 and irradiance of 40 mW/cm2 were the most effective irradiation parameters to stimulate and modulate oxidative stress in the primary teeth-derived dental pulp cells.


Subject(s)
Dental Pulp , Infrared Rays , Cell Survival , Oxidative Stress , Reactive Oxygen Species
2.
Clin Oral Investig ; 25(8): 4749-4779, 2021 Aug.
Article in English | MEDLINE | ID: mdl-34181097

ABSTRACT

OBJECTIVES: The aim of this review is to highlight recent progress in the field of biomaterials-mediated dental pulp tissue engineering. Specifically, we aim to underscore the critical design criteria of biomaterial platforms that are advantageous for pulp tissue engineering, discuss models for preclinical evaluation, and present new and innovative multifunctional strategies that hold promise for clinical translation. MATERIALS AND METHODS: The current article is a comprehensive overview of recent progress over the last 5 years. In detail, we surveyed the literature in regenerative pulp biology, including novel biologic and biomaterials approaches, and those that combined multiple strategies, towards more clinically relevant models. PubMed searches were performed using the keywords: "regenerative dentistry," "dental pulp regeneration," "regenerative endodontics," and "dental pulp therapy." RESULTS: Significant contributions to the field of regenerative dentistry have been made in the last 5 years, as evidenced by a significant body of publications. We chose exemplary studies that we believe are progressive towards clinically translatable solutions. We close this review with an outlook towards the future of pulp regeneration strategies and their clinical translation. CONCLUSIONS: Current clinical treatments lack functional and predictable pulp regeneration and are more focused on the treatment of the consequences of pulp exposure, rather than the restoration of healthy dental pulp. CLINICAL RELEVANCE: Clinically, there is great demand for bioinspired biomaterial strategies that are safe, efficacious, and easy to use, and clinicians are eager for their clinical translation. In particular, we place emphasis on strategies that combine favorable angiogenesis, mineralization, and functional tissue formation, while limiting immune reaction, risk of microbial infection, and pulp necrosis.


Subject(s)
Endodontics , Regenerative Endodontics , Biocompatible Materials , Dental Pulp , Humans , Lab-On-A-Chip Devices , Regeneration , Tissue Engineering
3.
J Prosthet Dent ; 124(1): 122.e1-122.e9, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32220399

ABSTRACT

STATEMENT OF PROBLEM: If the components in the acrylic resins used to fabricate interim crows are cytotoxic, they can interfere with the integrity of the adjacent periodontal tissue and the dentin-pulp complex. PURPOSE: The purpose of this in vitro study was to assess the cytotoxicity of resin-based materials used to prepare interim crowns. MATERIAL AND METHODS: The following materials were used in this study: CAR, conventional acrylic resin powder and liquid; BR, bis-acrylic resin; and PAR, pressed acrylic resin of the CAD-CAM type. Glass disks were used as the control (Co). Oral epithelial cells (NOK) were seeded on glass disks and standardized disks prepared with the resins under study. After incubation for 24 hours, the cells were analyzed for viability (Alamar Blue and Live or Dead), adhesion, and morphology (SEM and fluorescence), as well as epidermal growth factor synthesis (EGF-ELISA). The surface roughness (Ra) of test specimens was evaluated under a confocal microscope. The data were submitted to ANOVA and the Tukey HSD statistical tests (α=.05). RESULTS: The highest Ra value was observed in BR in comparison with CAR, PAR, and Co (P<.05). The highest viability, adhesion, and EGF synthesis values were determined for the cells in contact with PAR (P<.001). CONCLUSIONS: The computer-aided design and computer-aided manufacturing (CAD-CAM)-type resin favored adhesion, metabolism, and epithelial cell proliferation, and it was therefore considered cytocompatible.


Subject(s)
Acrylic Resins , Crowns , Composite Resins , Computer-Aided Design , Dental Materials , Materials Testing , Surface Properties
4.
J Endod ; 44(6): 971-976.e1, 2018 Jun.
Article in English | MEDLINE | ID: mdl-29703618

ABSTRACT

INTRODUCTION: The improvement of biomaterials capable of driving the regeneration of the pulp-dentin complex mediated by resident cells is the goal of regenerative dentistry. In the present investigation, a chitosan scaffold (CHSC) that released bioactive concentrations of simvastatin (SIM) was tested, aimed at the development of a cell-free tissue engineering system. METHODS: First, we performed a dose-response assay to select the bioactive dose of SIM capable of inducing an odontoblastic phenotype in dental pulp cells (DPCs); after which we evaluated the synergistic effect of this dosage with the CHSC/DPC construct. SIM at 1.0 µmol/L (CHSC-SIM1.0) and 0.5 µmol/L were incorporated into the CHSC, and cell viability, adhesion, and calcium deposition were evaluated. Finally, we assessed the biomaterials in an artificial pulp chamber/3-dimensional culture model to simulate the cell-free approach in vitro. RESULTS: SIM at 0.1 µmol/L was selected as the bioactive dose. This drug was capable of strongly inducing an odontoblastic phenotype on the DPC/CHSC construct. The incorporation of SIM into CHSC had no deleterious effect on cell viability and adhesion to the scaffold structure. CHSC-SIM1.0 led to significantly higher calcium-rich matrix deposition on scaffold/dentin disc assay compared with the control (CHSC). This biomaterial induced the migration of DPCs from a 3-dimensional culture to its surface as well as stimulated significantly higher expressions of alkaline phosphatase, collagen type 1 alpha 1, dentin matrix acidic phosphoprotein 1, and dentin sialophosphoprotein on 3-dimensional-cultured DPCs than on those in contact with CHSC. CONCLUSIONS: CHSC-SIM1.0 scaffold was capable of increasing the chemotaxis and regenerative potential of DPCs.


Subject(s)
Cell-Free System/drug effects , Chitosan/therapeutic use , Dental Pulp/physiology , Dentin/physiology , Regeneration/drug effects , Simvastatin/therapeutic use , Tissue Engineering/methods , Tissue Scaffolds , Cell-Free System/physiology , Cells, Cultured , Dose-Response Relationship, Drug , Humans , Male , Regenerative Endodontics/methods , Simvastatin/administration & dosage , Young Adult
5.
Acta Biomater ; 68: 190-203, 2018 03 01.
Article in English | MEDLINE | ID: mdl-29294374

ABSTRACT

In this study, we investigated the anti-inflammatory, odontogenic and pro-angiogenic effects of integrating simvastatin and nanofibrous poly(l-lactic acid) (NF-PLLA) scaffolds on dental pulp cells (DPCs). Highly porous NF-PLLA scaffolds that mimic the nanofibrous architecture of extracellular matrix were first fabricated, then seeded with human DPCs and cultured with 0.1 µM simvastatin and/or 10 µg/mL pro-inflammatory stimulator lipopolysaccharide (LPS). The gene expression of pro-inflammatory mediators (TNF-α, IL-1ß and MMP-9 mRNA) and odontoblastic markers (ALP activity, calcium content, DSPP, DMP-1 and BMP-2 mRNA) were quantified after long-term culture in vitro. In addition, we evaluated the scaffold's pro-angiogenic potential after 24 h of in vitro co-culture with endothelial cells. Finally, we assessed the combined effects of simvastatin and NF-PLLA scaffolds in vivo using a subcutaneous implantation mouse model. The in vitro studies demonstrated that, compared with the DPC/NF-PLLA scaffold constructs cultured only with pro-inflammatory stimulator LPS, adding simvastatin significantly repress the expression of pro-inflammatory mediators. Treating LPS+ DPC/NF-PLLA constructs with simvastatin also reverted the negative effects of LPS on expression of odontoblastic markers in vitro and in vivo. Western blot analysis demonstrated that these effects were related to a reduction in NFkBp65 phosphorylation and up-regulation of PPARγ expression, as well as to increased phosphorylation of pERK1/2 and pSmad1, mediated by simvastatin on LPS-stimulated DPCs. The DPC/NF-PLLA constructs treated with LPS/simvastatin also led to an increase in vessel-like structures, correlated with increased VEGF expression in both DPSCs and endothelial cells. Therefore, the combination of low dosage simvastatin and NF-PLLA scaffolds appears to be a promising strategy for dentin regeneration with inflamed dental pulp tissue, by minimizing the inflammatory reaction and increasing the regenerative potential of resident stem cells. STATEMENT OF SIGNIFICANCE: The regeneration potential of stem cells is dependent on their microenvironment. In this study, we investigated the effect of the microenvironment of dental pulp stem cells (DPSCs), including 3D structure of a macroporous and nanofibrous scaffold, the inflammatory stimulus lipopolysaccharide (LPS) and a biological molecule simvastatin, on their regenerative potential of mineralized dentin tissue. The results demonstrated that LPS upregulated inflammatory mediators and suppressed the odontogenic potential of DPSCs. Known as a lipid-lowing agent, simvastatin was excitingly found to repress the expression of pro-inflammatory mediators, up-regulate odontoblastic markers, and exert a pro-angiogenic effect on endothelial cells, resulting in enhanced vascularization and mineralized dentin tissue regeneration in a biomimetic 3D tissue engineering scaffold. This novel finding is significant for the fields of stem cells, inflammation and dental tissue regeneration.


Subject(s)
Dental Pulp/cytology , Inflammation/pathology , Nanofibers/chemistry , Odontogenesis/drug effects , Polyesters/chemistry , Simvastatin/pharmacology , Tissue Scaffolds/chemistry , Alkaline Phosphatase/metabolism , Biomarkers/metabolism , Cell Differentiation/drug effects , Cytoprotection/drug effects , Gene Expression Regulation/drug effects , Human Umbilical Vein Endothelial Cells/drug effects , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Lipopolysaccharides , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/metabolism , Neovascularization, Physiologic/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Signal Transduction/drug effects , Young Adult
6.
J Contemp Dent Pract ; 17(10): 861-866, 2016 Oct 01.
Article in English | MEDLINE | ID: mdl-27794160

ABSTRACT

AIM: This study evaluated the influence of acid-etching time on collagen exposure in adhesive interfaces established on primary and permanent dentin. MATERIALS AND METHODS: Flat dentin surfaces were produced on sound primary molars and premolars (n = 8). The surfaces were divided into mesial and distal halves, and each half was etched with phosphoric acid for 5 or 15 seconds. The teeth were randomly allocated into two groups according to the adhesive system applied: Prime & Bond NT or Prime & Bond 2.1. After the adhesive application, the specimens were processed for Goldner's trichrome staining. The thickness of the uninfiltrated collagen zone (UCZ) in the hybrid layer was measured under optical microscopy. Data were analyzed by analysis of variance and Tukey tests (α = 0.05). RESULTS: The thickness of UCZ was adhesive dependent. Within the same substrate, the specimens treated with Prime & Bond 2.1 presented thicker UCZ when etched for 15 seconds. Collagen exposure was significantly higher for the primary teeth etched for 5 seconds and treated with Prime & Bond 2.1. CONCLUSION: The thickness of UCZ in hybrid layers is directly affected by acid-etching time and by the adhesive system applied. Primary dentin seems to be more susceptible to collagen exposure than is permanent dentin. CLINICAL SIGNIFICANCE: Both acid-etching time and adhesive system can influence the amount of exposed collagen interfering on resin-dentin bond quality, especially on primary dentin.


Subject(s)
Acid Etching, Dental/methods , Collagen/ultrastructure , Dentition, Permanent , Tooth, Deciduous , Bicuspid/ultrastructure , Dental Bonding , Humans , Microscopy , Molar/ultrastructure , Phosphoric Acids , Time Factors
7.
Arch Oral Biol ; 60(9): 1428-36, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26163982

ABSTRACT

OBJECTIVE: To evaluate the effect of a 17.5% H2O2 gel on the odontoblastic differentiation capability of human dental pulp cells (HDPCs). DESIGN: The bleaching gel was applied for 45, 15 or 5min to enamel/dentine discs adapted to transwells, positioned over previously cultured HDPCs. In the control group, no treatment was performed on the discs. Immediately after samples were bleached, the cell viability (MTT assay) and death (Live/Dead assay) as well as the mRNA gene expression of inflammatory mediators (TNFα, IL-1ß, IL-6, and COX-2; real-time PCR) were evaluated. The mRNA gene expression of odontoblastic markers (DMP-1, DSPP, and ALP) and mineralized nodule deposition (alizarin red) were assessed at 7, 14 and 21 days post-bleaching. The amount of H2O2 in contact with cells was quantified. Data were evaluated by Kruskal-Wallis and Mann-Whitney tests (α=5%). RESULTS: Significant cell viability reduction and cell death were observed for bleached groups relative to control in a time-dependent fashion. Also, significant overexpression of all inflammatory mediators tested occurred in the 45- and 15-min groups. In the bleached groups, the expression of ALP, DMP-1, and DSPP and the deposition of mineralized nodules were reduced in comparison with those in the control group, at the initial periods (7 and 14 days). However, the 15- and 5-min groups reached values similar to those in the control group at the 21-day period. CONCLUSIONS: The 17.5% H2O2 gel was cytotoxic to pulp cells; however, cells subjected to short-term bleaching are capable of expressing the odontoblastic phenotype over time.


Subject(s)
Dental Enamel/drug effects , Dental Pulp/cytology , Hydrogen Peroxide/pharmacology , Odontoblasts/drug effects , Tooth Bleaching , Animals , Cattle , Cell Culture Techniques , Cell Differentiation/drug effects , Cell Survival/drug effects , Cells, Cultured , Gels , Gene Expression , Humans , In Vitro Techniques , Inflammation Mediators/metabolism , Microscopy, Fluorescence , Phenotype , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction
8.
J Dent ; 42(3): 351-8, 2014 Mar.
Article in English | MEDLINE | ID: mdl-24056046

ABSTRACT

OBJECTIVES: To evaluate the effects of experimental protocols on bleaching effectiveness and hydrogen peroxide (HP) diffusion through enamel and dentine. METHODS: Enamel/dentine discs were subjected to six bleaching sessions, consisting of 1 or 3 applications of 17.5% or 35%-HP gel for 5/15min, or 37% carbamide peroxide (CP) gel for 10/20min. Discs undergoing the regular protocol (35%-HP; 3×15min) constituted the positive control group. Colour change (ΔE) was assessed (CIE L*a*b* system) after each session. HP diffusion was quantified (sessions 1, 3, and 6) in enamel/dentine discs adapted to artificial pulp chambers. Data were analysed by Pillai's Trace and Bonferroni test, or by one-way ANOVA and SNK/Tamhane's test (α=5%). RESULTS: All tooth-bleaching protocols significantly increased the ΔE values. A reduction in HP diffusion and no significant difference in ΔE compared with the positive control were observed for the following bleaching protocols: 17.5%-HP 3×15min, at the 4th session; and 35%-HP 1×15 and 3×5min, at the 5th session. HP diffusion in the 37%-CP 3×20min bleaching protocol was statistically similar to that in the positive control. The other experimental bleaching protocols significantly decreased HP diffusion through enamel/dentine discs, but the ΔE values were statistically lower than those observed in the positive control, in all sessions. CONCLUSION: Shortening the contact time of a 35%-HP gel or reducing its concentration produces gradual tooth colour change and reduced HP diffusion through enamel and dentine. CLINICAL SIGNIFICANCE: A reduction in HP concentration, from 35% to 17.5%, in a bleaching gel or shortening its application time on enamel provides a significant tooth-bleaching improvement associated with decreased HP diffusion across hard dental tissues. Therefore, these protocols may be an interesting alternative to be tested in the clinical situation.


Subject(s)
Dental Enamel/metabolism , Dentin/metabolism , Hydrogen Peroxide/pharmacokinetics , Tooth Bleaching Agents/pharmacokinetics , Tooth Bleaching/methods , Animals , Carbamide Peroxide , Cattle , Color , Dental Enamel/drug effects , Dental Pulp Cavity/metabolism , Dentin/drug effects , Diffusion , Gels , Hydrogen Peroxide/administration & dosage , Materials Testing , Peroxides/administration & dosage , Peroxides/pharmacokinetics , Spectrophotometry/methods , Time Factors , Tooth Bleaching Agents/administration & dosage , Tooth Discoloration/drug therapy , Tooth Discoloration/metabolism , Urea/administration & dosage , Urea/analogs & derivatives , Urea/pharmacokinetics
9.
Photochem Photobiol ; 90(1): 201-6, 2014 01.
Article in English | MEDLINE | ID: mdl-23931553

ABSTRACT

The aim of this study was to evaluate the effect of low-level laser therapy (LLLT) on odontoblast-like cells exposed to a bleaching agent. Mouse dental papilla cell-23 cells were seeded in wells of 24-well plates. Eight groups were established according to the exposure to the bleaching agent and LLLT (0, 4, 10 and 15 J cm(-2) ). Enamel-dentin disks were adapted to artificial pulp chambers, which were individually placed in wells containing Dulbecco's modified Eagle's medium (DMEM). A bleaching agent (35% hydrogen peroxide [BA35%HP]) was applied on enamel (15 min) to obtain the extracts (DMEM + BA35%HP components diffused through enamel/dentin disks). The extracts were applied (1 h) to the cells, and then subjected to LLLT. Cell viability (Methyl tetrazolium assay), alkaline phosphatase (ALP) activity, as well as gene expression of ALP, fibronectin (FN) and type I collagen, were evaluated. The bleaching procedures reduced the cell viability, ALP activity and gene expression of dentin proteins. Laser irradiation did not modulate the cell response; except for FN, as LLLT decreased the gene expression of this protein by the cells exposed to the BA35%HP. It can be concluded that BA35%HP decreased the activities of odontoblasts that were not recovered by the irradiation of the damaged cells with low-level laser parameters tested.


Subject(s)
Bleaching Agents/pharmacology , Dental Pulp/drug effects , Lasers , Animals , Cell Line , Cell Survival/drug effects , Cell Survival/radiation effects , Mice , Real-Time Polymerase Chain Reaction , Tetrazolium Salts/pharmacology
10.
Gerontology ; 59(6): 534-41, 2013.
Article in English | MEDLINE | ID: mdl-23867757

ABSTRACT

BACKGROUND: Bisphosphonates are potent inhibitors of bone resorption. These kinds of drugs, which are used for the treatment of osteolytic diseases, have been associated with the occurrence of oral osteonecrosis, especially in patients over 60 years old. Current studies have demonstrated that the cytotoxic effects of bisphosphonates on osteoblasts play an important role in oral osteonecrosis development. OBJECTIVE: The aim of this study was to evaluate the effect of long-term application of a highly potent bisphosphonate - zoledronic acid (ZA) - on human osteoblasts in vitro. METHODS: Human osteoblasts (MG63 cell line) were seeded for 72 h in wells of 24-well plates. The Dulbecco's modified Eagle's medium (DMEM) was then replaced by culture medium without fetal bovine serum (FBS), and the cells were incubated for an additional 24 h, after which ZA was added to the DMEM without FBS and incubated in contact with osteoblasts for 7, 14 or 21 days. Cell viability (CV), total protein production (TPP), alkaline phosphatase (ALP) activity, mineral nodule formation (MNF), and gene expression of ALP and osteocalcin (OCN), as well as cell morphology by scanning electronic microscopy, were evaluated. Data were statistically analyzed by Kruskal-Wallis and Mann-Whitney tests, with a significance level of 5%. RESULTS: The cytotoxic effects of ZA on osteoblasts were characterized by reduction of CV, TPP, ALP and MNF production. In addition, ZA MNF caused a decrease in gene expression of ALP and OCN, as well as intense cell morphology alterations. All these negative effects of ZA were concentration and period dependent. CONCLUSION: Both concentrations of ZA (1 and 5 µM) caused cytotoxic effects to osteoblasts which reduced the production and expression of proteins that play an important role in bone matrix synthesis and mineralization.


Subject(s)
Bone Density Conservation Agents/toxicity , Diphosphonates/toxicity , Imidazoles/toxicity , Osteoblasts/drug effects , Alkaline Phosphatase/genetics , Alkaline Phosphatase/metabolism , Animals , Bisphosphonate-Associated Osteonecrosis of the Jaw/etiology , Bisphosphonate-Associated Osteonecrosis of the Jaw/genetics , Bisphosphonate-Associated Osteonecrosis of the Jaw/metabolism , Bone Matrix/drug effects , Bone Matrix/metabolism , Calcification, Physiologic/drug effects , Cattle , Cell Line , Cell Survival/drug effects , Culture Media, Serum-Free , Gene Expression/drug effects , Humans , Osteoblasts/metabolism , Osteoblasts/pathology , Osteocalcin/genetics , Osteocalcin/metabolism , Zoledronic Acid
11.
Int J Dent ; 2012: 719452, 2012.
Article in English | MEDLINE | ID: mdl-22844284

ABSTRACT

The aim of this study was to determine adequate energy doses using specific parameters of LLLT to produce biostimulatory effects on human gingival fibroblast culture. Cells (3 × 10(4) cells/cm(2)) were seeded on 24-well acrylic plates using plain DMEM supplemented with 10% fetal bovine serum. After 48-hour incubation with 5% CO(2) at 37°C, cells were irradiated with a InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm; 40 mW) with energy doses of 0.5, 1.5, 3, 5, and 7 J/cm(2). Cells were irradiated every 24 h totalizing 3 applications. Twenty-four hours after the last irradiation, cell metabolism was evaluated by the MTT assay and the two most effective doses (0.5 and 3 J/cm(2)) were selected to evaluate the cell number (trypan blue assay) and the cell migration capacity (wound healing assay; transwell migration assay). Data were analyzed by the Kruskal-Wallis and Mann-Whitney nonparametric tests with statistical significance of 5%. Irradiation of the fibroblasts with 0.5 and 3 J/cm(2) resulted in significant increase in cell metabolism compared with the nonrradiated group (P < 0.05). Both energy doses promoted significant increase in the cell number as well as in cell migration (P < 0.05). These results demonstrate that, under the tested conditions, LLLT promoted biostimulation of fibroblasts in vitro.

12.
Arch Oral Biol ; 57(2): 131-8, 2012 Feb.
Article in English | MEDLINE | ID: mdl-21920494

ABSTRACT

OBJECTIVES: To characterize the mechanical and biological properties of a resin-modified glass ionomer cement (RMGIC) containing doxycycline hyclate. METHODS: The antibacterial effect of RMGIC containing 1.5, 3.0 and 4.5% doxycycline hyclate was assessed using two experiments - agar diffusion test for 24h and biofilm assay for 24h and 7 days - against some cariogenic bacteria. Briefly, base layers of BHI agar and 300µL of each inoculum were prepared in Petri dishes with 6 wells that were completely filled with materials. After 24h incubation, zones of bacterial growth inhibition were measured using a digital caliper. Biofilm assays were conducted using RMGIC specimens immersed in 24-well plates containing the inoculum in BHI broth. After 24h and 7 days, each specimen were removed, vortexed and the suspension diluted and inoculated in BHI plates for subsequent bacterial counting. Cytotoxicity tests used 50 specimens made in sterilized metal molds, including Vitrebond as positive control. Extracts from every specimen were applied on the MDPC-23 odontoblast-like cells for 24h. The MTT assay and SEM evaluation determined cell metabolism and morphology, respectively. 80 cylindrical specimens were made from the previously cited groups, and were submitted to testing with a universal testing machine (Instron 4411) using a crosshead speed of 1.0mm/min for compressive strength and 0.5mm/min for diametral tensile strength, respectively. Data from antibacterial and cytotoxic effects, and mechanical properties were submitted to appropriated statistical tests. RESULTS: All tested groups showed growth inhibition of all tested strains (p<0.05) in 24h for both microbiological tests, but only 4.5% doxycycline have antibacterial effect after 7 days. None of doxycycline concentrations caused toxic effect to the MDPC-23 cells or presenting alterations to mechanical properties. CONCLUSION: The incorporation of up to 4.5% doxycycline hyclate into RMGIC inhibits important oral microorganisms, without modifying biological and mechanical characteristics of the dental material, suggesting a new alternative for the treatment of dental caries.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Dental Caries/drug therapy , Doxycycline/analogs & derivatives , Glass Ionomer Cements/chemistry , Resin Cements/chemistry , Anti-Bacterial Agents/pharmacology , Biological Assay , Cell Line , Colony Count, Microbial , Doxycycline/pharmacology , Doxycycline/therapeutic use , Microbial Sensitivity Tests
13.
Braz Dent J ; 22(6): 502-10, 2011.
Article in English | MEDLINE | ID: mdl-22189647

ABSTRACT

The aim of this study was to evaluate the effect of specific parameters of low-level laser therapy (LLLT) on biofilms formed by Streptococcus mutans, Candida albicans or an association of both species. Single and dual-species biofilms--SSB and DSB--were exposed to laser doses of 5, 10 or 20 J/cm(2) from a near infrared InGaAsP diode laser prototype (LASERTable; 780 ± 3 nm, 0.04 W). After irradiation, the analysis of biobilm viability (MTT assay), biofilm growth (cfu/mL) and cell morphology (SEM) showed that LLLT reduced cell viability as well as the growth of biofilms. The response of S. mutans (SSB) to irradiation was similar for all laser doses and the biofilm growth was dose dependent. However, when associated with C. albicans (DSB), S. mutans was resistant to LLLT. For C. albicans, the association with S. mutans (DSB) caused a significant decrease in biofilm growth in a dose-dependent fashion. The morphology of the microorganisms in the SSB was not altered by LLLT, while the association of microbial species (DSB) promoted a reduction in the formation of C. albicans hyphae. LLLT had an inhibitory effect on the microorganisms, and this capacity can be altered according to the interactions between different microbial species.


Subject(s)
Biofilms/radiation effects , Candida albicans/radiation effects , Lasers, Semiconductor , Low-Level Light Therapy/instrumentation , Mouth/microbiology , Streptococcus mutans/radiation effects , Bacteriological Techniques , Biofilms/growth & development , Candida albicans/growth & development , Candida albicans/ultrastructure , Coloring Agents , Dose-Response Relationship, Radiation , Humans , Hyphae/radiation effects , Materials Testing , Microbial Interactions/radiation effects , Microbial Viability/radiation effects , Microscopy, Electron, Scanning , Mycology/methods , Radiation Dosage , Streptococcus mutans/growth & development , Streptococcus mutans/ultrastructure , Succinate Dehydrogenase/analysis , Temperature , Tetrazolium Salts , Thiazoles , Time Factors
14.
Eur J Oral Sci ; 118(4): 411-6, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20662916

ABSTRACT

The aim of this study was to evaluate the mechanical stability of resin-dentin bonds produced in vivo in the presence of chlorhexidine. Children presenting at least one pair of contralateral primary molars with occlusal carious lesions were enrolled in the study. After cavity preparation and phosphoric acid etching, dentin was treated with 5 microl of either 2% chlorhexidine (experimental group) or deionized water (control group) [corrected].Thirteen pairs of teeth were restored and were collected after physiological exfoliation. The periods in oral function after restoration were divided as follows: up to 30 d; and 1-5, 10-12, and 18-20 months. Beam-shaped specimens (0.81 mm(2)) were obtained and subjected to microtensile bond-strength testing. A significant decrease of the bond strength was observed in the control group starting at the 1-5 month period (30.6%), while in the experimental group this decrease started only after 10-12 months of function (26.3%). The use of chlorhexidine as an adjuvant to the adhesion to dentin did not produce any detrimental effect to the immediate bond strength and was capable of reducing the rate of resin-dentin bond degradation within the first few months after restoration.


Subject(s)
Chlorhexidine/chemistry , Dental Bonding , Dentin-Bonding Agents/chemistry , Dentin/ultrastructure , Enzyme Inhibitors/chemistry , Resin Cements/chemistry , Acid Etching, Dental , Child , Composite Resins/chemistry , Dental Caries/therapy , Dental Cavity Preparation/methods , Dental Materials/chemistry , Dental Restoration, Permanent , Dental Stress Analysis/instrumentation , Follow-Up Studies , Humans , Materials Testing , Matrix Metalloproteinase Inhibitors , Molar/pathology , Organophosphonates/chemistry , Polymethacrylic Acids/chemistry , Stress, Mechanical , Tensile Strength , Time Factors , Tooth, Deciduous/pathology
15.
Dent Mater ; 22(10): 954-62, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16387355

ABSTRACT

OBJECTIVE: The aim of this study was to evaluate the pulp response following cementation of inlays using two different resin cements. METHODS: Deep Class V cavities were prepared on the buccal surface of 34 sound human premolars. Impressions were taken and inlays were prepared which were cemented with the following luting materials-Group 1: Rely X Unicem (3M ESPE); Group 2: Variolink II (Ivoclar Vivadent). In Group 3 (control), after lining the cavity floor with Dycal (Dentsply Caulk) the inlays were cemented with Rely X Unicem. Four additional teeth were used as an intact control group. For Variolink II, the adhesive system Excite was used as part of the cementation procedure. After 7 or 60 days, the teeth were extracted and processed for histological assessment. RESULTS: At 7 days, Rely X Unicem and Variolink II system triggered in two samples a mild and moderate inflammatory response, respectively. At 60 days, the pulpal response decreased for both groups. A discrete persistent inflammatory response occurred in Group 2 in which displacement of resin components across the dentin tubules was observed. In the control group, normal histological characteristics were observed. The inflammatory response and tissue disorganization were related to the remaining dentin thickness between the cavity floor and the pulp tissue. SIGNIFICANCE: Techniques for inlay cementation using distinct luting cements may cause specific pulpal damage. Variolink II associated with the adhesive system Excite cause more aggressive effects to the pulp-dentin complex than Rely X Unicem cement when both are used to cement inlay restorations.


Subject(s)
Dental Pulp/drug effects , Methacrylates/toxicity , Resin Cements/toxicity , Adolescent , Cementation/adverse effects , Child , Dentin/anatomy & histology , Humans , Inlays
16.
Dent Mater ; 22(9): 864-9, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16388848

ABSTRACT

OBJECTIVE: The aim of this in vitro study was to evaluate the cytotoxicity of resin-modified glass-ionomer lining cements submitted to different curing regimes and applied to an immortalized odontoblast-cell line (MDPC-23). METHODS: Forty round-shaped specimens of each experimental material (Fuji Lining LC and Vitrebond) were prepared. They were light-cured for the manufacturers' recommended time (MRT = 30 s), under-cured (0.5 MRT = 15 s), over-cured (1.5 MRT = 45 s) or allowed to dark cure (0 MRT). Sterilized filter papers soaked with either 5 microL of PBS or HEMA were used as negative and positive control, respectively. After placing the specimens individually in wells of 24-well dishes, odontoblast-like cells MDPC-23 (30,000 cells/cm2) were plated in each well and incubated for 72 h in a humidified incubator at 37 degrees C with 5% CO2 and 95% air. The cytotoxicity was evaluated by the cell metabolism (MTT assay) and cell morphology (SEM). RESULTS: Fuji Lining LC was less cytotoxic than Vitrebond (p < 0.05) in all the experimental conditions. However, the cytotoxicity of Fuji Lining LC was noticeably increased in the absence of light-curing while the same was not observed for Vitrebond. The length of light-curing (15, 30 or 45 s) did not influence the toxicity of both lining materials when they were applied on the odontoblast-cell line MDPC-23. SIGNIFICANCE: The light-activation plays an important role in reducing the cytotoxicity of Fuji Lining LC. Following the manufacturer' recommendation regarding the light-curing regime may prevent toxic effect to the pulp cells.


Subject(s)
Dentin-Bonding Agents/toxicity , Glass Ionomer Cements/toxicity , Odontoblasts/drug effects , Resins, Synthetic/toxicity , Animals , Cell Line, Transformed , Cell Shape/drug effects , Coloring Agents/metabolism , Dental Cavity Lining , Dentin-Bonding Agents/radiation effects , Glass Ionomer Cements/chemistry , Glass Ionomer Cements/radiation effects , Light , Methacrylates/toxicity , Mice , Microscopy, Electron, Scanning , Phase Transition , Resins, Synthetic/radiation effects , Tetrazolium Salts/metabolism
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