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Cavity disinfection becomes an important step before a dental restorative procedure. The disinfection can be obtained cleaning the dental cavity with antimicrobial agents before the use of adhesive systems. The aim of this study was to conduct a systematic review on the effect of different cavity disinfectants on restorations’ adhesion and clinical success. A search was carried out through the Cochrane Library, PubMed, and Web of Science. In vitro and in situ studies reporting results on dentin bond strength tests, and clinical studies published until August 2020, in English, Spanish and Portuguese were included. The methodological quality assessment of the clinical studies was carried out using the Revised Cochrane risk-of-bias tool. Chlorhexidine could preserve adhesion to dentin. EDTA and ethanol had positive results that should be further confirmed. Given the significant lack of scientific evidence, the use of lasers, fluoridated agents, sodium hypochlorite, or other products as cavity disinfectants should be avoided. Chlorhexidine is a safe option for cavity disinfection with adequate preservation of adhesion to dentin. Moreover, future researches should be focused on the efficacy of these disinfectants against cariogenic bacteria and their best application methods.

The present study aimed to investigate the correlation between chronological age and pulp–tooth volume ratio in anterior teeth using cone beam computed tomographic (CBCT) images and provide equations for age estimation based on pulp–tooth volume ratio. In this cross-sectional study, CBCT images of 312 anterior teeth of the maxilla and mandible were examineαd. The chronological age and the sex of the patients were recorded. Tooth volume and pulp volume of the anterior teeth were measured using Mimics software, and then tooth–pulp volume ratio was calculated. Statistical analysis of data was performed using chi-square test, independent T-test, Pearson’s correlation, and linear regression (α = 0.05). Finally, equations were made based on the results of the regression analysis for age estimation in general and for males and females. The age of the subjects ranged between 16 and 69 years (mean 40.6 ± 12.74). The pulp–tooth volume ratio of the maxillary teeth was generally higher than the mandibular teeth. A significant inverse relationship between age and pulp–tooth volume ratio was observed for all anterior teeth. (p &lt, 0.05). The strongest correlation between age and pulp–tooth volume ratio was reported for mandibular central incisor (r = −0.58, p &lt, 0.001) and the weakest was for mandibular lateral incisor (r = −0.36, p = 0.012). Age estimation was most accurate among females using the maxillary central incisors (p &lt, 0.001) and among males using the mandibular central incisors and maxillary canines (p = 0.003). Pulp–tooth volume ratio of mandibular central incisors, maxillary lateral incisors, and maxillary canines in males and maxillary and mandibular central incisors, maxillary lateral incisors, and maxillary and mandibular canines in females can be used for age estimation.

The implementation of hydraulic calcium silicate-based endodontic cements (HCSCs) in biologically based endodontic procedures for the primary dentition has been recently investigated, focusing on the biological response of stem cells from human exfoliated deciduous teeth (SHEDs) towards them. The present systematic review aimed to present a qualitative synthesis of the available literature consisting of in vitro assays, which assessed the cytocompatibility and bioactive properties of HCSCs in direct contact with SHEDs. Following the PRISMA statement, an electronic database search was carried out in Medline, Scopus, Embase, Web of Science, and SciELO on March 31st and updated on November 16th, 2020. In vitro studies evaluating the biological response of SHEDs to the treatment with HCSCs were eligible. Within the term biological response, assays assessing the cytocompatibility (i.e., cell viability, migration, proliferation), cell plasticity or differentiation (i.e., osteo/odontogenic marker expression), and bioactivity or biomineralization (i.e., mineralized nodule formation) were included. A total of seven studies were included after the selection process. The study sample comprised an extensive range of cell viability, migration, proliferation, adhesion, and bioactivity assays regarding the biological response of SHEDs towards five different commercially available HCSCs (MTA, ProRoot MTA, Biodentine, iRoot BP Plus, and Theracal LC). Biodentine, MTA, and iRoot BP Plus showed significant positive results in cytocompatibility and bioactivity assays when cultured with SHEDs. The results from in vitro assays assessing the cytocompatibility and bioactivity of the HCSCs MTA, Biodentine, and iRoot BP Plus towards SHEDs support their use in vital pulp treatment for the primary dentition.

Researchers have always been interested in finding new and effective materials for protection against radiation. This experimental study aimed to design and fabricate new types of nano-material and micro-material based shields against the ionizing effect of cone beam computed tomography (CBCT) X-rays. To fabricate a flexible prototype, we added dioctyl phthalate (DOP) oil to emulsion polyvinyl chloride (PVC) powder. The paste was mixed and dispersed. Then, nano- and micro-powders of WO3 and Bi2O3 were added to the paste, with the weight ratio of 20% PVC, 20% DOP, and 60% nano- and micro-metals. Using an ultrasonic mixer, the polymer matrix and metals were mixed and a paste with a thick texture was developed. The resultant paste was poured into glass molds and the molds were then heated in an oven. After cooling, the resultant sheets were selected for further experiments. A CBCT unit and dosimeter were used to evaluate the characterization and X-ray shielding properties of the fabricated prototypes. The half-value layers (HVL) for nano-WO3, micro-WO3, nano-Bi2O3, and micro-Bi2O3 were 0.0390, 0.0524, 0.0351, and 0.0374 cm, respectively. In addition, the linear attenuation coefficient (&micro, ) for these materials were 17.77, 13.20, 19.71, and 18.5 cm&minus, 1, respectively. The findings indicate that nano-structured samples are more effective in the attenuation of X-ray energy. The nano-structured WO3 prototype was nearly 34% more efficient in attenuating radiation compared to the micro-structured WO3 prototype. This difference in nano- and micro-structured Bi2O3 prototypes was 6.5%.

Biocompatibility is an essential property for any vital pulp material that may interact with the dental pulp tissues. Accordingly, this study aimed to compare the chemical composition and ultrastructural morphology of Biodentine (Septodont, Saint Maur-des-Fosses, France), ProRoot MTA (Dentsply Tulsa Dental Specialties, Johnson City, TN, USA), and Bio-C Repair (Angelus, Londrina, PR, Brazil), as well as their biological effects on human dental pulp cells. Chemical element characterization of the materials was undertaken using scanning electron microscopy and energy dispersive X-ray analysis (SEM-EDX). The cytotoxicity was assessed by analyzing the cell viability (MTT assay), cell morphology (immunofluorescence assay), and cell attachment (flow cytometry assay). The results were statistically analyzed using ANOVA and Tukey&rsquo, s test (p &lt, 0.05). EDX revealed that ProRoot MTA and Biodentine were mostly composed of calcium, carbon, and oxygen (among others), whereas Bio-C Repair evidenced a low concentration of calcium and the highest concentration of zirconium. SEM showed adequate attachment of human dental pulp cells (hDPCS) to vital pulp materials and cytoskeletal alterations were not observed in the presence of material eluates. Remarkably, the undiluted Biodentine group showed higher viability than the control group cells (without eluates) at 24 h, 48 h, and 72 h (p &lt, 0.001). Based on the evidence derived from an in vitro cellular study, it was concluded that Bio-C Repair showed excellent cytocompatibility that was similar to Biodentine and ProRoot MTA.

Aim: The purpose of the study was the evaluation of the esthetic and physical changes produced on colored titanium Grade 5 (Ti6Al4V) laser treated surfaces to be used in implant dentistry for esthetic success. Materials and methods: Colored titanium surfaces were obtained with laser treatment. The physical and topographic properties were evaluated by stereo, light, and electron microscopy and profilometric analyses. L*a*b* colorimetric coordinates were measured by spectrometry, and the superficial chemical characteristics were evaluated by energy dispersive X-ray analysis. Results: Within the complete palette of titanium colors, pinks (P1-P2), incarnadine (I), and white (W) obtained by laser were selected. The topography, texture, hues, saturation, roughness, and porosity of the samples were compared with those of machined (M) and sand-blasted and etched (SBAE) control surfaces. P1, P2, and I, similar in hue and roughness (Ra @ 0.5 &mu, m), had a microgroove spacing of 56 &mu, m and a decreasing porosity. The W sample with a &ldquo, checkerboard&rdquo, texture and a light color (L* 96.31) was similar to the M samples (Ra = 0.32 &mu, m), but different from SBAE (Ra = 1.41 &mu, m, L* 65.47). Discussion: The aspects of hard and soft tissue could result in an esthetic failure of the dental implant by showing the dark color of the fixture or abutment. The two different pinks and incarnadine surfaces showed favorable esthetic and physical features to promote dental implant success even in the maxillary anterior area with gingival recession, asymmetry, and deficiency. Conclusion: Titanium colored laser surfaces represent a valid alternative to those currently traditionally obtained and interesting and potential perspectives in the management of dental implants&rsquo, esthetic failure.

New bioactive materials have been developed for retrograde root filling. These materials come into contact with vital tissues and facilitate biomineralization and apical repair. The objective of this study was to evaluate the cytocompatibility and bioactivity of two bioactive cements, Bio-C Repair (Angelus, Londrina, Pr, Brazil) and TotalFill BC RRM putty (FGK, Dentaire SA, La-Chaux-de-fonds, Switzerland). The biological properties in human periodontal ligament stem cells (hPDLSCs) that were exposed to Bio-C Repair and TotalFill BC RRM putty were studied. Cell viability, migration, and cell adhesion were analyzed. Moreover, qPCR and mineralization assay were performed to evaluate the bioactivity potential of these cements. The results were statistically analyzed using ANOVA and the Tukey test (p &lt, 0.05). It was observed that cell viability and cell migration in Bio-C Repair and TotalFill BC RRM putty were similar to the control without statistically significant differences, except at 72 h when TotalFill BC RRM putty was slightly lower (p &lt, 0.05). Excellent cell adhesion and morphology were observed with both Bio-C Repair and TotalFill BC RRM putty. Both cements promoted the osteo- and cementogenic differentiation of hPDLSCs. These results suggest that Bio-C Repair and TotalFill BC RRM putty are biologically appropriate materials to be used as retrograde obturation material.

Background and Objective: Metabolic syndrome (MetS) is associated with an increased risk of periodontitis even if the mechanism is unknown. Since both MetS and periodontitis are characterized by an alteration of inflammation status, the aim of this pilot study was to determine if differences in ROS metabolism of phagocytes isolated from (A) patients with MetS, (B) patients with both MetS and mild periodontitis, (C) healthy subjects and (D) normal weight subjects with mild periodontitis, were present. Methods: ROS metabolism was studied by a Chemiluminescence (CL) technique: the system was made up of luminol (100 nmol/L) and cells (1 × 105) in the presence or absence of stimulus constituted by opsonized zymosan (0.5 mg). The final volume (1.0 mL) was obtained using modified KRP buffer. ROS production was measured at 25°C for 2 h, using an LB 953 luminometer (Berthold, EG & G Co, Germany). All the experiments were performed in triplicate. Statistical Analysis: All results are mean ± standard deviation (SD). The group of means was compared by the analysis of variance "(ANOVA)". A value of p < 0.05 was considered significant. Results: Results showed that basal ROS production (both from PMNs and from PBMs) of groups A, B and D was increased with respect to that obtained from group C (p Conclusion: These results are congruent with literature data, although the actual clinical relevance of the phenomenon remains to be evaluated.

The aim of this study was to investigate the effect of orthodontic resins on cell survival and to evaluate monomer leaching both before and after resin polymerization. Materials and methods: 3T3 mouse fibroblasts were exposed to three cured and uncured orthodontic resins. Cellular viability was assessed by Alamar Blue assay after 24, 48 and 72 h. High Performance Liquid Chromatography was used to measure the amount of monomers released by the tested samples. Data were analyzed by means of ANOVA and Tukey’s test (p

The bone-implant interface influences peri-implant bone healing and osseointegration. Among various nano-engineering techniques used for titanium surface modification, anodization is a simple, high-throughput and low-cost process, resulting in a nanoporous oxide coating which can promote osseointegration and impart antimicrobial and immunomodulatory properties. We anodized rounded tip dental implants of commercial grade titanium in aqueous phosphoric acid modified with calcium and potassium acetate, and characterized the resulting surface morphology and composition with scanning electron microscopy and energy dispersive spectrometry. The appearance of nanopores on these implants confirmed successful nanoscale morphology modification. Additionally, the metal cations of the used salts were incorporated into the porous coating together with phosphate, which can be convenient for osseointegration. The proposed method for surface nanostructuring of titanium alloy could allow for fabrication of dental implants with improved biocompatibility in the next stage of research.

Human periapical cyst mesenchymal stem cells (hPCy-MSCs) are a newly discovered cell population innovatively collected from inflammatory periapical cysts. The use of this biological waste guarantees a source of stem cells without any impact on the surrounding healthy tissues, presenting a valuable potential in tissue engineering and regenerative medicine applications. In the present study, hPCy-MSCs were collected, isolated, and seeded on three experimental mineral-doped porous scaffolds produced by the thermally-induced phase-separation (TIPS) technique. Mineral-doped scaffolds, composed of polylactic acid (PLA), dicalcium phosphate dihydrate (DCPD), and/or hydraulic calcium silicate (CaSi), were produced by TIPS (PLA-10CaSi, PLA-5CaSi-5DCPD, PLA-10CaSi-10DCPD). Micro-CT analysis evaluated scaffolds micromorphology. Collected hPCy-MSCs, characterized by cytofluorimetry, were seeded on the scaffolds and tested for cell proliferation, cells viability, and gene expression for osteogenic and odontogenic differentiation (DMP-1, OSC, RUNX-2, HPRT). Micro-CT revealed an interconnected highly porous structure for all the scaffolds, similar total porosity with 99% open pores. Pore wall thickness increased with the percentage of CaSi and DCPD. Cells seeded on mineral-doped scaffolds showed a superior proliferation compared to pure PLA scaffolds (control), particularly on PLA-10CaSi-10DCPD at day 12. A higher number of non-viable (red stained) cells was observable on PLA scaffolds at days 14 and 21. DMP-1 expression increased in hPCy-MSCs cultured on all mineral-doped scaffolds, in particular on PLA-5CaSi-5DCPD and PLA-10CaSi-10DCPD. In conclusion, the innovative combination of experimental scaffolds colonized with autologous stem cells from periapical cyst represent a promising strategy for regenerative healing of periapical and alveolar bone. info:eu-repo/semantics/publishedVersion

The aim of the study was to investigate the effects of several decalcifying agents used as irrigant solutions in endodontic treatment on collagen and mineral components of dentin. Coronal dentin discs from five caries-free human third molars with a smear layer were treated for one minute with a chelating solution (1% Ethylenediaminetetraacetic acid (EDTA), 10% EDTA, 17% EDTA, 10% citric acid). Mineralization degree (Ca/N and P/N atomic ratios, IR Iapatite/Iamide II and I1410(carbonate)/I554(phosphate) spectroscopic ratios) and possible collagen rearrangements (collagen infrared (IR) amide II e III shifts) were evaluated by environmental scanning electron microscopy (ESEM)/energy dispersive X-ray spectroscopy (EDX) and IR spectroscopy before and after treatment (T0) and after ageing (T24h and T2m) in simulated body fluid (SBF). At T0, analysis showed that the highest demineralizing effect was achieved using a 10% citric acid solution and 10% EDTA, while the smallest effect was observed when using 17% EDTA. No significant collagen modifications were detected upon treatment with 1% EDTA, while subtle changes were observed after the other treatments. At T24h or T2m, analyses showed the highest remineralization values for 1% EDTA and the lowest for 10% citric acid, mainly at T2m. The samples treated with 17% EDTA showed slight collagen rearrangements upon remineralization. In conclusion, the highest demineralizing effect was observed for 10% EDTA and 10% citric acid. Collagen rearrangement was found for all the treatments except for 1% EDTA. The highest remineralization capability in SBF values was recorded for 1% EDTA and the lowest for 10% citric acid. A slight collagen rearrangement upon remineralization was still present in 17% EDTA-treated samples. Clinical use as a chelating agent in the endodontic therapy of citric acid and concentrated EDTA solutions should be reconsidered.

Dentinal tubule penetration influences root canal treatment sealing. The aim of this study was to compare dentinal penetration of two clinical presentations of silicate-based sealers using confocal laser. Sixty single-rooted human teeth from 50–70 year-old patients extracted for orthodontic/periodontal reasons were used. Canals were prepared using Mtwo system up to 35/0.04, with 5.25% NaOCl irrigation and final irrigation using 17% EDTA. Teeth were randomly assigned into study groups (ni = 20): EndoSequence BC sealer (ES, group 1), BioRoot RCS (BR, group 2); and a control group (nc = 20) with AH Plus (AHP). Root canals were obturated with 35/0.04 gutta-percha (single-cone technique). The samples were obtained from apical, middle, and coronal thirds. Dentinal tubule penetration depth and percentage of penetration around the canal perimeter were measured. The statistical analysis was performed using Mann Whitney U test and Wilcoxon t-test (95% confidence interval). ES exhibited a significantly higher penetration than AHP in apical and middle thirds (p < 0.05), and in middle and coronal thirds relative to BR (p < 0.05). The percentage of penetration around the canal perimeter was significantly higher for ES compared to BR in all thirds, but only in the apical third for AHP (p < 0.05). The pre-mixed silicate-based sealer exhibited better penetration than the powder/liquid one.

L-Carnitine is an amino acid derivative that plays a key role in the metabolism of fatty acids, including the shuttling of long-chain fatty acyl CoA to fuel mitochondrial β-oxidation. In addition, L-carnitine reduces oxidative damage and plays an essential role in the maintenance of cellular energy homeostasis. L-carnitine also plays an essential role in the control of cerebral functions, and the aberrant regulation of genes involved in carnitine biosynthesis and mitochondrial carnitine transport in Drosophila models has been linked to neurodegeneration. Drosophila models of neurodegenerative diseases provide a powerful platform to both unravel the molecular pathways that contribute to neurodegeneration and identify potential therapeutic targets. Drosophila can biosynthesize L-carnitine, and its carnitine transport system is similar to the human transport system; moreover, evidence from a defective Drosophila mutant for one of the carnitine shuttle genes supports the hypothesis of the occurrence of β-oxidation in glial cells. Hence, Drosophila models could advance the understanding of the links between L-carnitine and the development of neurodegenerative disorders. This review summarizes the current knowledge on L-carnitine in Drosophila and discusses the role of the L-carnitine pathway in fly models of neurodegeneration.

A reverse phase high performance liquid chromatographic (RP-HPLC) method was developed for identification and estimation of 18-β-glycyrrhetinic acid (GA) in HepG2 cell line. The analysis was carried out using a JASCO HPLC system with a C-18 (3 μm) Supelco reversed phase column (150 x 4.7 mm) using a mobile phase of 80% CH3OH and 20% of CH3CN: tetrahydrofuran: water (10:80:10, v/v/v). The method was linear in the concentration range of 1.5–120 μg /mL (n = 5). The LOD and LOQ were determined based on standard deviation of the y-intercept and the slope of the calibration curve. The LOD and LOQ values were found to be 11.46 μg/mL and 34.72 μg/mL, respectively. The mean percentage recovery by standard addition experiments of GA is 92.4 % ± 5.2%. The intracellular GA concentration value, obtained as mean of five different determinations, was 45.8 ± 7.45 μg/mL. We have developed a HPLC-UV method for quantitative determination of GA inside cells, with advantages in the cost reduction and economy of the analytical process.

Over the last three decades, it has been frequently reported that the properties of dental restorative composites cured with argon laser are similar or superior to those achieved with conventional halogen and light emitting diode (LED) curing units. Whereas laser curing is not dependent on the distance between the curing unit and the material, such distance represents a drawback for conventional curing units. However, a widespread clinical application of this kind of laser remains difficult due to cost, heavy weight, and bulky size. Recently, with regard to the radiation in the blue region of the spectrum, powerful solid-state lasers have been commercialized. In the current research, CAD (computer-aided design)/CAM (computer-aided manufacturing) assisted solid-state lasers were employed for curing of different dental restorative composites consisting of micro- and nanoparticle-reinforced materials based on acrylic resins. Commercial LED curing units were used as a control. Temperature rise during the photopolymerisation process and bending properties were measured. By providing similar light energy dose, no significant difference in temperature rise was observed when the two light sources provided similar intensity. In addition, after 7 days since curing, bending properties of composites cured with laser and LED were similar. The results suggested that this kind of laser would be suitable for curing dental composites, and the curing process does not suffer from the tip-to-tooth distance.

Purpose The aim of this study was to evaluate the effects of various prophylactic treatments of titanium implants on bacterial biofilm formation, correlating surface modifications with the biofilms produced by Pseudomonas aeruginosa PAO1, Staphylococcus aureus, and bacteria isolated from saliva. Methods Pure titanium disks were treated with various prophylactic procedures, and atomic force microscopy (AFM) was used to determine the degree to which surface roughness was modified. To evaluate antibiofilm activity, we used P. aeruginosa PAO1, S. aureus, and saliva-isolated Streptococcus spp., Bacteroides fragilis, and Staphylococcus epidermidis. Results AFM showed that the surface roughness increased after using the air-polishing device and ultrasonic scaler, while a significant reduction was observed after using a curette or polishing with Detartrine ZTM (DZ) abrasive paste. In addition, we only observed a significant (P, Graphical Abstract

Introduction The literature reveals controversies regarding the formation of para-chloroaniline (PCA) when chlorhexidine (CHX) is mixed with sodium hypochlorite (NaOCl). This study aimed to investigate the stability of PCA in the presence of NaOCl and to examine the in vitro cytotoxic effects of CHX/NaOCl reaction mixtures. Methods Different volumes of NaOCl were added to CHX (mix 1) or PCA (mix 2). Upon centrifugation, the supernatant and precipitate fractions collected from samples were analyzed using high-performance liquid chromatography. The cytotoxic effects of both fractions were examined on human periodontal ligament and 3T3 fibroblast cell lines. Results High-performance liquid chromatographic analysis showed no PCA signal when NaOCl was mixed with CHX (mix 1). In mix 2, the intensity of PCA was decreased when NaOCl was added to PCA, and chromatographic signals, similar to that of CHX/NaOCl, were also observed. The mortality of precipitates exerted on both cell lines was lower compared with that of supernatants. Conclusions The discrepancy in the data from the literature could be caused by the instability of the PCA in the presence of NaOCl. The CHX/NaOCl reaction mixture exhibits a wide range of cytotoxic effects.

Background: Titanium dental implants are today widely used with osseointegration mainly dependently on the implant surface properties. Different processing routes lead to different surface characteristics resulting, of course, in different in situ behaviors of the implants. Materials: The effect of different treatments, whether mechanical or chemical, on the surface morphology of titanium implants were investigated. To this aim, various experimental methods, including roughness analysis as well scanning electron microscope (SEM) observations, were applied. Results: The results showed that, in contrast to the mechanical treatments, the chemical ones gave rise to a more irregular surface. SEM observations suggested that where commercial pure titanium was used, the chemical treatments provided implant surfaces without contaminations. In contrast, sandblasted implants could cause potential risks of surface contamination because of the presence of blasting particles remnants. Conclusions: The examined implant surfaces showed different roughness levels in relation to the superficial treatment applied. The acid-etched surfaces were characterized by the presence of deeper valleys and higher peaks than the sandblasted surfaces. For this reason, acid-etched surfaces can be more easily damaged by the stress produced by the peri-implant bone during surgical implant placement.

Dental composite resins are biomaterials commonly used to aesthetically restore the structure and function of teeth impaired by caries, erosion, or fracture. Residual monomers released from resin restorations as a result of incomplete polymerization processes interact with living oral tissues. Monomers like triethylene glycol dimethacrylate (TEGDMA) or 2-hydroxylethyl methacrylate (HEMA) are cytotoxic via apoptosis, induce genotoxic effects, and delay the cell cycle. Monomers also influence the response of cells of the innate immune system, inhibit specific odontoblast cell functions, or delay the odontogenic differentiation and mineralization processes in pulp-derived cells including stem cells. These observations indicate that resin monomers act as environmental stressors which inevitably disturb regulatory cellular networks through interference with signal transduction pathways. We hypothesize that an understanding of the cellular mechanisms underlying these phenomena will provide a better estimation of the consequences associated with dental therapy using composite materials, and lead to innovative therapeutic strategies and improved materials being used at tissue interfaces within the oral cavity. Current findings strongly suggest that monomers enhance the formation of reactive oxygen species (ROS), which is most likely the cause of biological reactions activated by dental composites and resin monomers. The aim of the present review manuscript is to discuss adaptive cell responses to oxidative stress caused by monomers. The particular significance of a tightly controlled network of non-enzymatic as well as enzymatic antioxidants for the regulation of cellular redox homeostasis and antioxidant defense in monomer-exposed cells will be addressed. The expression of ROS-metabolizing antioxidant enzymes like superoxide dismutase (SOD1), glutathione peroxidase (GPx1/2), and catalase in cells exposed to monomers will be discussed with particular emphasis on the role of glutathione (GSH), which is the major non-enzymatic antioxidant. The causal relationship between vital cell functions like the regulation of cell survival or cell death in monomer-treated cell cultures and the availability of GSH will be highlighted. We will also consider the influence of monomer-induced oxidative stress on central signal transduction pathways including mitogen-activated protein kinases (MAPK) ERK1/2, p38, and JNK as well as the stress-activated transcription factors downstream Elk-1, ATF-2, ATF-3, and cJun. Finally, we address signaling pathways originating from monomer-induced DNA damage including the activation of ATM (ataxia-telangiectasia mutated), Chk2, p53, p21, and H2AX. The understanding of the mechanisms underlying adaptive cell responses will stimulate a constructive debate on the development of smart dental restorative materials which come into contact with oral tissues and effective strategies in dental therapy.

Background: Auxiliary power sources (LED and laser) are used in in-office teeth bleaching techniques to accelerate the redox reaction of the whitening gel to increase ease of use, to improve comfort and safety, and to decrease the procedure time. Objective: The aim this study is to evaluate the efficiency of the teeth whitening procedures performed with hydrogen peroxide and carbamide peroxide, LED or Laser activated. Method: 18 patients, affected by exogenous dyschromia, were treated with a bleaching agent composed by 35% hydrogen peroxide and 10% carbamide peroxide. They were divided into two groups: in the first group the bleaching agent was activated by a LED lamp; in the second group it was activated by a Laser diode lamp. Both groups were subjected to 3 bleaching cycle of 15’ each. The chromatic evaluations were performed before and after one week from the treatment, using a chromatic scale and a spectrophotometer. The mean value of pre, post bleaching and follow-up were analyzed using a T-test, with results statistically significant for P Results: Results showed that the variations in brightness, chroma and hue are significantly influenced by the interaction between the whitening agent and the original colour of the teeth. Laser-activation has marginally improved the bleaching effectiveness. All patients treated with laser activation complained an increase in dental sensitivity. Conclusion: The use of laser-activating systems did not improve the efficacy of bleaching.

Resin monomers like 2-hydroxyethyl methacrylate (HEMA) disturb cell functions including responses of the innate immune system, mineralization and differentiation, or induce cell death via apoptosis. These phenomena are associated with oxidative stress and a reduction in the concentration of the antioxidant glutathione (GSH), resulting in imbalanced redox homeostasis. Thus far, the precise mechanism of how resin monomers interfere with cellular redox regulation is unknown. The present study provides insight into the induction of apoptosis and the differential expression of antioxidant enzymes depending on the availability of GSH. Buthionine sulfoximine (BSO) was used to inhibit GSH synthesis, while 2-oxothiazolidine-4-carboxylate (OTC), and N-acetylcysteine (NAC) as prodrugs supported GSH synthesis in RAW264.7 mouse macrophages exposed to HEMA (0-8 mm) for 24 h. The level of GSH was significantly decreased after cells were preincubated with BSO, and the formation of reactive oxygen species (ROS) increased in cultures subsequently exposed to HEMA. Apoptosis was drastically increased by BSO in HEMA-exposed cell cultures as well, but OTC and NAC retracted HEMA-induced cell death. These results show that dental monomer-induced apoptosis is causally related to the availability of GSH. The hydrogen peroxide decomposing enzymes glutathione peroxidase (GPx1/2) and catalase were differentially regulated in HEMA-exposed cultures. Expression of GPx1/2 was inhibited by HEMA and further reduced in the presence of BSO. SOD1 (superoxide dismutase) expression was inhibited in the presence of HEMA, and was decreased to an even greater extent by BSO, possibly due to H(2)O(2)-feedback inhibition. The expression of catalase was considerably up-regulated in HEMA-exposed cultures, implying that H(2)O(2) is the type of ROS that is significantly increased in monomer-exposed cells. OTC and NAC counteracted the effect of HEMA on GPx1/2, SOD1, and catalase expression. HO-1 (heme oxygenase) expression was strongly enhanced by HEMA, suggesting the need for further antioxidants like bilirubin to support enzyme activities that directly regulate H(2)O(2) equilibrium. Expression of the oxidoreductase thioredoxin (TRX1), the second major thiol-dependent antioxidant system in eukaryotic cells, was slightly reduced, while the oxygen-sensing protein HIF-1α was downregulated in HEMA-exposed cell cultures. These results indicate that cells and tissues actively respond to monomer-induced oxidative stress by the differential expression of enzymatic antioxidants.

Several in vitro studies have reported contrasting values for triethylene glycol-dimethacrylate (TEGDMA) concentrations shown to induce cytotoxic effects. The aim of this study was to evaluate the effective concentrations of TEGDMA reached under the routine experimental conditions used in biocompatibility in vitro tests and determines changes in cytotoxicity and the associated production of reactive oxygen species (ROS) based on different TEGDMA solutions. TEGDMA was added to cell culture medium either directly or previously dissolved in dimethyl sulfoxide (DMSO) or ethanol (EtOH), both in the presence and absence of cells. Intracellular and extracellular TEGDMA concentrations were determined by high performance liquid chromatography (HPLC). The cytotoxicity effects of TEGDMA preparations were determined in 3T3-fibroblasts by 3-(4,5 dimethyiazol-2-1)-2-5-diphenyl tetrazolium bromide assay. The production of ROS was measured by flow cytometry. In the absence of cells the effective final TEGDMA concentrations obtained in Dulbecco's Modified Eagle Medium were significantly lower than the nominal one. When 2 mmol/L TEGDMA was first solubilized in DMSO or EtOH, a significant decrease in cell viability, and an increase in ROS productioncompared to pure TEGDMAwas observed. After 2 h of incubation, TEGDMA previously dissolved in DMSO or ETOH was reduced by 15% and 20%, respectively, whereas otherwise it remained unaffected. Our results demonstrate that the effective concentration of TEGDMA dissolved in culture medium (in the presence or absence of solvents) does not concur with the nominal one. Therefore, the presence of the utilized solvents does not substantially alter the monomer solubility but eases its entrance into the cells thus improving its cytotoxic potency. (c) 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 100B: 15001506, 2012.

The production of PEEA microspheres with potential as carriers for protein oral delivery is described. PEEAs with different hydrophilicity were synthesized and characterized. Experiments showed that an increase in copolymer hydrophilicity gave particles less prone to cell interaction. BSA release profiles from PEEA microspheres demonstrated that an increase in polymer hydrophilicity was useful in limiting protein burst and modulating drug delivery rate by increasing PEEA degradability. These results show that fine-tuning of the hydrophilic/hydrophobic properties of PCL is essential for the formulation protein-loaded microspheres with specific properties.

Objectives Dental resin monomers like triethylene glycol dimethacrylate (TEGDMA) and 2-hydroxyethyl methacrylate (HEMA) are able to cause an imbalance of the redox state in mammalian cells. The resulting oxidative stress originating from reactive oxygen species (ROS) has been associated with cytotoxicity. We hypothesized that ROS might contribute to the generation of genotoxicity by TEGDMA and HEMA as well. Therefore, we examined the formation of micronuclei in V79 cells by both resin monomers in the presence of the antioxidant N-acetylcysteine (NAC), which scavenges ROS. In addition, we analyzed the effects of TEGDMA and HEMA on the normal cell cycle in the presence of NAC. Methods V79 fibroblasts were exposed to increasing concentrations of TEGDMA and HEMA in the presence and absence of NAC for 24 h. Genotoxicity was indicated by the formation of micronuclei. The modification of the normal cell cycle was analyzed by flow cytometry (FACS). Results A dose-related increase in the number of micronuclei in V79 cells-induced by TEGDMA and HEMA indicated genotoxicity of both chemicals. However, the formation of micronuclei was reduced in the presence of 10 mmol/L NAC, indicating its protective role. A cell cycle delay in G2 phase caused by TEGDMA was absent when cells were co-treated with NAC. Similarly, the presence of NAC led to a reversion of the cell cycle delay in HEMA-treated cell cultures. Significance Our results suggest that genotoxic effects and the modification of the cell cycle caused by TEGDMA and HEMA are mediated, at least in part, by oxidative stress.

Monomers are released from dental resin materials, and thus cause adverse biological effects in mammalian cells. Cytotoxicity and genotoxicity of some of these methacrylates have been identified in a vast number of investigations during the last decade. It has been well-established that the co-monomer triethylene glycol dimethacrylate (TEGDMA) causes gene mutations in vitro. The formation of micronuclei is indicative of chromosomal damage and the induction of DNA strand breaks detected with monomers like TEGDMA and 2-hydroxyethyl methacrylate (HEMA). As a consequence of DNA damage, the mammalian cell cycle was delayed in both G1 and G2/M phases, depending on the concentrations of the monomers. Yet, the mechanisms underlying the genetic and cellular toxicology of resin monomers have remained obscure until recently. New findings indicate that increased oxidative stress results in an impairment of the cellular pro- and anti-oxidant redox balance caused by monomers. It has been demonstrated that monomers reduced the levels of the natural radical scavenger glutathione (GSH), which protects cell structures from damage caused by reactive oxygen species (ROS). Depletion of the intracellular GSH pool may then significantly contribute to cytotoxicity, because a related increase in ROS levels can activate pathways leading to apoptosis. Complementary, cytotoxic, and genotoxic effects of TEGDMA and HEMA are inhibited in the presence of ROS scavengers like N-acetylcysteine (NAC), ascorbate, and Trolox (vitamin E). Elevated intracellular levels of ROS can also activate a complex network of redox-responsive macromolecules, including redox-sensitive transcription factors like nuclear factor kappaB (NF-κB). It has been shown that NF-κB is activated probably to counteract HEMA-induced apoptosis. The induction of apoptosis by TEGDMA in human pulp cells has been associated with an inhibition of the phosphatidylinositol 3-kinase (PI3-K) cell-survival signaling pathway. Although the details of the mechanisms leading to cell death, genotoxicity, and cell-cycle delay are not completely understood, resin monomers may be able to alter the functions of the cells of the oral cavity. Pathways regulating cellular homeostasis, dentinogenesis, or tissue repair may be modified by monomers at concentrations well below those which cause acute cytotoxicity.

Several studies have already demonstrated that the incomplete polymerization of resin-based dental materials causes the release of monomers which might affect cell metabolism. The aim of this study was to investigate the effects of triethylene glycol dimethacrylate, 1,4-butanediol dimethacrylate, urethane dimethacrylate, and 2-hydroxyethyl methacrylate on (1) cellular energy metabolism, evaluating oxygen consumption rate, glucose consumption, glucose 6-phosphate dehydrogenase activity, and lactate production, and (2) cellular redox status, through the evaluation of glutathione concentration and of the activities of enzymes regulating glutathione metabolism.Methods. Human pulp cells were used and oxygen consumption was measured by means of a Clark electrode. Moreover, reactive oxygen species production was quantified. Enzymatic activity and glucose and lactate concentrations were determined through a specific kit.Results. Triethylene glycol dimethacrylate, 1,4-butanediol dimethacrylate, and 2-hydroxyethyl methacrylate induced a decrease in oxygen consumption rate, an enhancement of glucose consumption, and lactate production, whilst glucose 6-phosphate dehydrogenase and glutathione reductase activity were not significantly modified. Moreover, the monomers induced an increase of reactive oxygen species production with a consequent increase of superoxide dismutase and catalase enzymatic activities. A depletion of both reduced and total glutathione was also observed.Conclusion. The obtained results indicate that dental monomers might alter energy metabolism and glutathione redox balance in human pulp cells.

Objective: To evaluate the effects of intraoral aging on surface properties of esthetic and conventional nickel-titanium (NiTi) archwires. Materials and Methods: Five NiTi wires were considered for this study (Sentalloy, Sentalloy High Aesthetic, Superelastic Titanium Memory Wire, Esthetic Superelastic Titanium Memory Wire, and EverWhite). For each type of wire, four samples were analyzed as received and after 1 month of clinical use by an atomic force microscope (AFM) and a scanning electronic microscope (SEM). To evaluate sliding resistance, two stainless steel plates with three metallic or three monocrystalline brackets, bonded in passive configuration, were manufactured; four as-received and retrieved samples for every wire were pulled five times at 5 mm/min for 1 minute by means of an Instron 5566, recording the greatest friction value (N). Data were analyzed by one-way analysis of variance and by Student's t-test. Results: After clinical use, surface roughness increased considerably. The SEM images showed homogeneity for the as-received control wires; however, after clinical use esthetic wires exhibited a heterogeneous surface with craters and bumps. The lowest levels of friction were observed with the as-received Superelastic Titanium Memory Wire on metallic brackets. When tested on ceramic brackets, all the wires exhibited an increase in friction (t-test; P < .05). Furthermore, all the wires, except Sentalloy, showed a statistically significant increase in friction between the as-received and retrieved groups (t-test; P < .05). Conclusion: Clinical use of the orthodontic wires increases their surface roughness and the level of friction.

Objective: To compare the surface roughness of different orthodontic archwires. Materials and Methods: Four nickel-titanium wires (Sentalloy®, Sentalloy® High Aesthetic, Titanium Memory ThermaTi Lite®, and Titanium Memory Esthetic®), three β-titanium wires (TMA®, Colored TMA®, and Beta Titanium®), and one stainless-steel wire (Stainless Steel®) were considered for this study. Three samples for each wire were analyzed by atomic force microscopy (AFM). Three-dimensional images were processed using Gwiddion software, and the roughness average (Ra), the root mean square (Rms), and the maximum height (Mh) values of the scanned surface profile were recorded. Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test (P < .05). Results: The Ra, Rms, and Mh values were expressed as the mean ± standard deviation. Among as-received archwires, the Stainless Steel (Ra = 36.6 ± 5.8; Rms = 48 ± 7.7; Mh = 328.1 ± 64) archwire was less rough than the others (ANOVA, P < .05). The Sentalloy High Aesthetic was the roughest (Ra = 133.5 ± 10.8; Rms = 165.8 ± 9.8; Mh = 949.6 ± 192.1) of the archwires. Conclusions: The surface quality of the wires investigated differed significantly. Ion implantation effectively reduced the roughness of TMA. Moreover, Teflon®-coated Titanium Memory Esthetic was less rough than was ion-implanted Sentalloy High Aesthetic.

Aim To evaluate the effects of repeated autoclave sterilization cycles on surface topography of conventional nickel–titanium ( NiTi ) and titanium nitride ( TiN )-coated rotary instruments. Methodology A total of 60 NiTi rotary instruments, 30 ProTaper (Dentsply Maillefer) and 30 TiN -coated AlphaKite (Komet/Gebr. Brasseler), were analysed. Instruments were evaluated in the as-received condition and after 1, 5 and 10 sterilization cycles. After sterilization, the samples were observed using scanning electron microscope (SEM), and surface chemical analysis was performed on each instrument with energy dispersive X-ray spectroscopy (EDS). Moreover, the samples were analysed by atomic force microscopy (AFM), and roughness average (Ra) and the root mean square value (RMS) of the scanned surface profiles were recorded. Data were analysed by means of anova followed by Tukey's test. Result Scanning electron microscope observations revealed the presence of pitting and deep milling marks in all instruments. EDS analysis confirmed that both types of instruments were composed mainly of nickel and titanium, whilst AlphaKite had additional nitride. After multiple autoclave sterilization cycles, SEM examinations revealed an increase in surface alterations, and EDS values indicated changes in chemical surface composition in all instruments. Ra and RMS values of ProTaper significantly increased after 5 (P = 0.006) and 10 cycles (P = 0.002) with respect to the as-received instruments, whilst AlphaKite showed significant differences compared with the controls after 10 cycles (P = 0.03). Conclusions Multiple autoclave sterilization cycles modified the surface topography and chemical composition of conventional and TiN -coated NiTi rotary instruments.

Objective: Metal alloys used in dentistry and in other biomedical fields may release nickel ions in the oral environment. The release of nickel might influence the normal biological and physiological processes, including tissue wound healing, cell growth and proliferation. The aim of this study was to evaluate in vitro the effects of nickel ions on cell cycle, viability and proliferation. Materials and Methods: Human osteosarcoma cells (U2OS) and human keratinocytes (HaCat) were exposed to different nickel chloride (NiCl2) concentrations (0 - 5mM) for various periods exposure. The viability of cultured cells was estimated by flow cytometry using Annexin V-FITC and Propidium Iodide (PI). Cell proliferation was evaluated by using carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and flow cytometry. Finally, the effects of NiCl2 on cell cycle were assessed and quantified by flow cytometry. Statistical analysis was performed by means of ANOVA followed by Tukey’s test. Results: NiCl2 induced a dose and time dependent decrease in cell viability. After 24h, 1mM NiCl2 caused a similar and significant reduction of viability in U2OS and HaCat cells, while higher NiCl2 concentrations and longer exposure times showed a reduced cytotoxic effect in HaCat as compared to U2OS cells. Exposure to NiCl2 caused a dose- and time-dependent inhibition of cell proliferation in both cell lines tested, with a prominent effect on U2OS cells. Furthermore, both cell lines exposed to NiCl2 exhibited significant changes in cell cycle distribution after 24h exposure 2mM NiCl2, as compared to untreated cells (p Conclusion: Our results indicate that release of nickel ions may affect cell proliferation. The inhibition of cell growth by NiCl2 is mediated by both cell cycle arrest and by induction of cell death.

Introduction Mineral trioxide aggregate (MTA) is known to stimulate the hard tissue repair process. The purpose of this study was to evaluate the ability of MTA to support the adhesion, proliferation, and migration of human bone marrow–derived mesenchymal stem cells (hMSCs). Methods White ProRoot MTA and white Portland cement were mixed and left to set 24 hours. MSCs were cultured on the samples and observed after 24 hours by confocal laser scanning microscopy (CLSM) by using the cytoskeleton marker CellTracker. Cell proliferation was evaluated by means of alamar blue assay in the presence and absence of differentiation medium during a period of 28 days, and cells seeded on polystyrene culture wells were the control. To assess the effect on migratory ability of hMSCs, a transwell migration assay was performed for 18 hours, positioning MTA and Portland cement in 6-well plates and the cells in 8-μm pore inserts. Results hMSCs observed under CLSM showed attachment and spread activity on the upper surface of the MTA. Cell proliferation was significantly higher on MTA than on Portland cement. A rate proliferation increase of the MTA group compared with the control was observed after 14 days in presence of basic medium, whereas the same effect was reached after 21 days in presence of differentiation medium. Moreover, MTA was able to enhance cell migration significantly more than Portland cement. Conclusions Our findings suggest that MTA was able to assist hMSC adhesion, growth, and migration.