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Abstract Several glycoconjugate-based vaccines against bacterial infections have been developed and licensed for human use. Polysaccharide (PS) analysis and characterization is therefore critical to profile the composition of polysaccharide-based vaccines. For PS content quantification, the majority of Ultra High Performance Liquid Chromatography (UHPLC) methods rely on the detection of selective monosaccharides constituting the PS repeating unit, therefore requiring chemical cleavage and tailored development: only a few methods directly quantify the intact PS. The introduction of charged aerosol detector (CAD) technology has improved the response of polysaccharide analytes, offering greater sensitivity than other detector sources (e.g., ELSD). Herein, we report the development of a universal UHPLC-CAD method (UniQS) for the quantification and quality evaluation of polysaccharide antigens (e.g., Streptococcus Pneumoniae, Neisseria meningitidis and Staphylococcus aureus). This work laid the foundation for a universal UHPLC-CAD format that could play an important role in future vaccine research and development helping to reduce time, efforts, and costs.

The Italian earthquake waveform data are collected here in a dataset suited for machine learning analysis (ML) applications. The dataset consists of nearly 1.2 million three-component (3C) waveform traces from about 50 000 earthquakes and more than 130 000 noise 3C waveform traces, for a total of about 43 000 h of data and an average of 21 3C traces provided per event. The earthquake list is based on the Italian Seismic Bulletin (http://terremoti.ingv.it/bsi, last access: 15 February 2020​​​​​​​) of the Istituto Nazionale di Geofisica e Vulcanologia between January 2005 and January 2020, and it includes events in the magnitude range between 0.0 and 6.5. The waveform data have been recorded primarily by the Italian National Seismic Network (network code IV) and include both weak- (HH, EH channels) and strong-motion (HN channels) recordings. All the waveform traces have a length of 120 s, are sampled at 100 Hz, and are provided both in counts and ground motion physical units after deconvolution of the instrument transfer functions. The waveform dataset is accompanied by metadata consisting of more than 100 parameters providing comprehensive information on the earthquake source, the recording stations, the trace features, and other derived quantities. This rich set of metadata allows the users to target the data selection for their own purposes. Much of these metadata can be used as labels in ML analysis or for other studies. The dataset, assembled in HDF5 format, is available at http://doi.org/10.13127/instance (Michelini et al., 2021).

Statement of Problem: Three-dimensional (3D) additive manufacturing (AM) is an evolving technology in dentistry, proposed as an alternative to subtractive milling manufacture (MM) or conventional processing. However, a systematic review of the use of AM technology instead of milling or conventional processing is lacking., Purpose: The purpose of this systematic review and meta-analysis was to evaluate the mechanical properties of 3D printed prosthetic materials compared with MM and conventional techniques., Material and Methods: An electronic search of the literature was conducted on the MEDLINE (via PubMed), Scopus, and Web of Science databases. The inclusion criteria were in vitro studies published in the last 5 years, in English or Italian, and with 3D AM printed dental prosthetic materials. Data extraction was focused on dental prosthetic materials (ceramics, polymers, and metals) and their mechanical properties: flexural strength, fracture load, hardness, roughness, removable partial denture (RPD) fit accuracy, trueness, marginal discrepancy, and internal fit. Data considered homogenous were subjected to meta-analysis using the Stata17 statistical software program (95% confidence interval [CI]; α=.05). Since all variables were continuous, the Hedge g measure was calculated. A fixed-effects model was used for I 2 =0%, while the statistical analysis was conducted using a random-effects model with I 2 >0%., Results: From a total of 3624 articles, 2855 studies were selected, and 76 studies included after full-text reading. The roughness of AM-printed ceramics generally increased compared with that of conventional processing while the marginal discrepancy was comparable both for ceramics and polymers. The flexural strength, hardness, and fracture load of AM-printed polymers were statistically lower than those of the conventional group (P<.05). No significant difference was detected in terms of hardness, roughness, marginal discrepancy, fracture load, trueness, or internal fit between the AM and MM techniques (P>.05). Milling techniques showed significantly higher values of flexural strength (Hedge g=-3.88; 95% CI, -7.20 to -0.58; P=.02), also after aging (Hedge g=-3.29; 95% CI, -6.41 to -0.17; P=.04), compared with AM printing., Conclusions: AM is comparable with MM in terms of mechanical properties, in particular with polymeric materials. The flexural strength of AM-printed prostheses is lower than with conventional and MM techniques, as are the parameters of hardness and fracture load, while the marginal discrepancy is similar to that of MM and conventional techniques. AM prostheses are commonly used for interim crowns and fixed partial dentures, as their rigidity and fracture resistance cannot support mastication forces for extended periods. More comparative studies are needed., (Copyright © 2022 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Outer membrane vesicles (OMVs) have been widely explored to develop vaccine candidates for bacterial pathogens due to their ability to combine adjuvant properties with immunogenic activity. OMV expresses a variety of proteins and carbohydrate antigens on their surfaces. For this reason, there is an analytical need to thoroughly characterize the species expressed at their surface: we here present a simple and accurate reversed-phase ultrahigh-performance liquid chromatography (RP-UPLC) method developed according to quality by design principles. This work provides an analytical alternative to the classical sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) characterization. The higher selectivity and sensitivity of the RP-UHPLC assay allow for the identification of additional protein species with respect to SDS-PAGE and facilitate its precise relative abundance quantification. According to validation results, the assay showed high accuracy, linearity, precision, repeatability, and a limit of quantification of 1% for less abundant proteins. This performance paves the way for improved production campaign consistency while also being analytically simple (no sample pretreatment required), making it suitable for routine quality control testing. In addition, the applicability of the assay to a wider range of vesicle classes (GMMA) was demonstrated., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Aim: Public and private health services, which provide both preventive and health promotion interventions, were forced to suddenly stop their activities to limit the risk of infections during the pandemic emergency. Oral health administration, including that of children, was affected by these planned medical service closures, from both therapeutic and preventive perspectives. This study aims to analyse the consequences, at the oral cavity level, of failures to treat patients of childhood age, considering the impact of carious pathology on quality of life and incorrect eating and oral hygiene habits, which may occur in this age group., Methods: This is a cross-sectional, single-center, observational study. One hundred patients from the Odontostomatological University Center (C.O.U.) of Perugia were randomly enrolled., Conclusion: Oral health status of the examined sample is satisfactory overall, considering the clinic's interruption of treatments with the resulting long period of no follow-up and the emotional and economic stress generated by the pandemic condition for both the young patients and their caregivers.

Aim: Parents and caregivers, particularly in Italy, often have limited knowledge about their children's oral health. The primary objective of the study is to evaluate the educational effectiveness of a book on nutrition and prevention of oral diseases entitled "Oral health of mother and child in the first 1000 days of life"., Materials: The sample for this study was composed of 103 adult Italian women who were potential caregivers of one or more children (e.g., mothers, grandmothers, babysitters, and educators). The enrolled women completed a preliminary online survey which included questions about their socio-demographic characteristics and their knowledge on oral health in the first 1000 days of life of newborns (30 questions). Following the survey, they received the educational book. After reading it, the participants completed a second online survey with the same 30 questions, to measure any improvement in their knowledge., Conclusion: It appears that our educational book about nutrition and prevention of oral diseases was effective in enhancing knowledge among the participants in our study. These findings suggest that this educational resource has the potential to be a valuable tool in preventing oral diseases in paediatric populations. However, further confirmation of these results should be obtained through randomised controlled trials.

Objective: To investigate the biological effects of electronic cigarette (e-cigarette) and heated tobacco product extracts respect to tobacco smoke extract on human gingival fibroblasts and human oral keratinocytes analysing cell viability, morphology, migration, apoptosis, cell cycle and epithelial-mesenchymal transition (EMT)., Design: Human gingival fibroblasts and human oral keratinocytes viability was analysed by MTT assay, cell morphology using scanning electron microscope (SEM) and cell migration by Scratch assay, a method that mimics the cell migration during wound healing in vivo. Apoptosis and cell cycle were analysed with flow cytometry and the related-gene expression of TP53, BCL2, CDKN2A and CDKN1A was indagated using real-time polymerase chain reaction. EMT process was analysed through expression of specific markers: CDH1, SNAI2, TWIST1, MMP2, FN1 and VIM. All investigations were evaluated after 24 h an in vitro exposure., Results: Undiluted tobacco smoke extract induced significant inhibition of cell viability and cell migration, caused morphological alterations and induced an increase in cell death. No alterations or damage were observed after treatment with e-cigarette extracts. Heated tobacco product extract induced proliferation as highlighted by an increase of cell viability, cell migration and alterations of cycle analysis., Conclusions: Comparing the different cigarette extracts, tobacco smoke turns out to be the most harmful, e-cigarette did not determine morphological and functional alterations and heated tobacco product must be carefully investigated for its possible clinical effects on oral cell populations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

GlaxoSmithKline (GSK) is currently developing a fully liquid presentation to ease the administration of the licensed quadrivalent conjugate vaccine (Menveo) against meningococcal serogroup A, C, W, and Y (MenACWY) infections. Herein, we report a new method for determining the free saccharide (FS) content of CRM 197 -MenACWY conjugated antigens, with the aim of improving accuracy and reproducibility. Mathematical models have been used to support technical knowledge in reducing the need for experimental development. This results in an improved, faster, and platform-based technique for FS separation with one single pretreatment applicable to all antigens of the multivalent meningococcal vaccine., Competing Interests: The authors declare the following competing financial interest(s): All the authors are employees of the GSK group of companies. Francesco Berti is listed as an inventor on patents owned by the GSK group of companies., (© 2022 The Authors. Published by American Chemical Society.)

GSK is currently working to improve the commercial presentation of the licensed quadrivalent conjugate vaccine (Menveo) for use against meningococcal serogroup A, C, W, Y (MenACWY) infections. Menveo consists of a primary, lyophilized vial, containing the serogroup A antigen that is reconstituted with the content of a second, liquid, vial that contains the serogroup C, W, Y antigens, to give the final liquid MenACWY product. Since the MenA structure is prone to hydrolytic degradation in liquid formulations, we used mathematical models to rationally design a clinical Phase 2 development plan and provide end of shelf-life (EoSL) and release specification setting for the MenACWY liquid product. By using development and clinical stability data, statistical models were built and used to predict both the MenA free saccharide (FS) and O-Acetyl (OAc) content during long-term storage conditions at 5 °C and stressed (accelerated) stability studies at 15 °C, 22.5 °C, 25 °C, 37 °C and 50 °C. This approach allowed us to define an aging plan for the clinical material to reach at least the required levels of MenA FS and OAc levels at product EoSL. The clinical material was then exposed to a temperature of 22.5 ± 2.5 °C for 59 days to generate FS OAc content of about 35% and 40%, respectively, which was then delivered to the patients in the clinical trial. To the best of our knowledge, this work represents the first example in the field of vaccine research where statistical models have been used to rationally design tailored lots, with the goal of setting EoSL and release specification limits based on data collected on artificially aged clinical material, in which the FS and OAc levels tested were intended to support a product shelf-life of at least 24 months., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Several glycoconjugate vaccines have been licensed or are currently in clinical development to prevent bacterial infections. Here we report the development of a single analytical assay to quantify the conjugated saccharide content, as alternative to two separated total and free (unconjugated) saccharide assays used so far, for a quadrivalent conjugate vaccine containing meningococcal serogroup A polysaccharide (α-1,6-linked N-acetylmannosamine phosphate repeating unit partly O-acetylated at position C 3 or C 4 ) coupled with CRM 197 protein. The results confirm a high linear correlation among the two approaches (conjugated saccharide content vs. difference of total saccharide and free saccharide). Conjugated saccharide content estimation is therefore demonstrated to be a suitable method to monitor the product quality of vaccines containing meningococcal serogroup A conjugate antigen, in the final filled presentation as demonstrated here and potentially on the bulk conjugate before formulation., (Copyright © 2022. Published by Elsevier Ltd.)

Aim: When treating patients with Autism Spectrum Disorder (ASD) the doctor-patient relationship can be very challenging. The dentist is often forced to work under general anaesthesia or conscious sedation. Children with ASD are patients with an increased risk of caries due to poor oral hygiene, a cariogenic diet and the use of xerostomal drugs. In this work therapeutic strategies used to treat this kind of patients are evaluated and a new method to treat children with ASD is presented in order to increase awareness about this condition in the dental field., Methods: The Atlas Center (a non-profit organisation in Peurgia, Italy) has developed a software, called paINTeraction, that allows these special children to immerse themselves in a virtual reality with the help of an operator. Through this system the child can explore the dental office (and all its sounds and noises) before the real dental visit, thus connecting to the dental professional, achieving greater compliance and reducing anxiety., Conclusion: paINTeraction, with the use of digital technology tools, may be particularly well suited to introduce patients to the therapeutic environment, particularly in the dental setting.

Aim: Malocclusions are widespread all over the world with high prevalence values and represent a social vulnerability and health problem because of the important burden of orthodontic treatment for both families and the public health service. The Italian Ministry of Health identified the principles for priority setting in dental care, i.e. patients with IOTN (Index of Orthodontic Treatment Need) 4-5 and those in particular conditions of health or social vulnerability., Methods: A retrospective study was carried out to assess the IOTN and malocclusion in a population attending the public Orthodontic Health Service of Perugia (Italy) from September 2018 to February 2020. Medical charts, social income information, study models, photos and lateral cephalograms were examined to assess the orthodontic treatment need., Conclusion: Orthodontic health service in Italy tends to have many patients on the waiting list, while the 72% of subject under orthodontic treatment are not part of the orthodontic priority (IOTN grade 4-5). This study suggests the heath system to guarantee a priority of need for orthodontic treatment for patients with health vulnerabilities.

Dental caries and periodontal disease represent a health problem and a social cost for the entire population, and in particular for socio-economically disadvantaged individuals who are less resistant to disease. The aim of this review is to estimate the prevalence and severity of the two dental pathologies, caries and periodontal disease, in the different classes of socio-economically disadvantaged subjects and to understand which of them are most affected. A systematic search of the literature was performed in MEDLINE (via PubMed), EMBASE and Web of Science after establishing a suitable search strategy for each database, using keywords related to socio-economically vulnerable classes and health outcomes. Socio-economically disadvantaged individuals are more susceptible to tooth decay and periodontal disease (with relative tooth loss) than non-vulnerable people. Additionally, when multiple vulnerabilities are combined in the same subject, these oral diseases worsen. There is no type of vulnerability more affected by caries and periodontitis than others, since overall they all have severe disease indices. The data from this systematic literature review might be useful for health policy makers looking to allocate more resources and services to socially disadvantaged individuals, resulting in making them more resilient to oral disease due to their social marginalization.

Objectives: The aim of this work is to investigate the biological effects of IQOS smoking on human gingival fibroblasts and human keratinocytes analysing cell viability, morphology, migration, apoptosis and cell cycle., Background: Electronic cigarettes and tobacco heating systems have been marketed to reduce smoking damages caused by combustion., Methods: Human gingival fibroblasts and human keratinocytes viability was determined by a colorimetric assay measuring mitochondrial dehydrogenase activity (MTT assay); after an in vitro exposure of 24 h, cell morphology was analysed with scanning electron microscope and cell migration was tested by Scratch assay, a method to mimic the migration of the cells during wound healing in vivo. Apoptosis and cell cycle were analysed with flow cytometry, and the expression of related genes (p53, Bcl2, p16 and p21) was indagated using real-time polymerase chain reaction., Results: IQOS extracts increased both cell viability (23%-41% with fibroblasts and 30%-79% with keratinocytes) and migration. No morphological alterations were observed. IQOS extracts did not induced an increase in cell death, but rose the number of S- and G2/M-phase cells. IQOS extracts also significantly increased p53 expression by fibroblasts (undiluted and 6.25% dilution, 2- and 3.6-fold higher, respectively) and reduced both Bcl2 (about two- and fivefold, respectively) and p21 expressions (about twofold with both extracts), while on keratinocytes both undiluted and 6.25% dilution extracts increased Bcl2 expression (about four- and threefold higher, respectively) and reduced p53 expression (about two- and fivefold, respectively)., Conclusion: IQOS smoke seemed to induce proliferation as highlighted by a viability assay, and migration and cell cycle analysis. The increased cell proliferation induced by IQOS devices must be carefully investigated for its possible clinical effects on oral cell populations., (© 2021 The Authors. Journal of Periodontal Research published by John Wiley & Sons Ltd.)

The aim of this study is to investigate the Erbium:Yttrio-Aluminum-Granate (Er:YAG) laser photothermal and mechanical effects on cariogenic species concentration and on the microbial load composition of therapeutic cavities, in order to evaluate the possible micro-organisms reduction and make a comparison with manual and rotating conventional therapy (CT). A clinical trial was designed, including adults with active deep carious lesions on permanent teeth who were divided into two groups, i.e., control group and intervention group treated with CT and Er:YAG therapy, respectively. Before and after any conservative treatment, two oral samples were collected using a small sterile microbrush scrubbed within the base of the dentinal cavity tissue. The percentage of reduction and the colony-forming units (CFUs) count after Er:YAG and conventional treatments were compared for total microorganisms, including Candida spp., Streptococcus spp., and Lactobacillus spp. The microbial reduction varied from 90.2% to 100% and was significantly observed for total microorganisms and Streptococcus spp. ( p < 0.05). The Er:YAG laser shows the potential for clinical applications, especially with paediatric and complicated patients, thanks to its minimally invasive properties and its effect on the reduction of microbial load.

To analyze the effects of four universal adhesives (Optibond Solo Plus-OB, Universal Bond-UB, Prime&Bond Active-PBA, FuturaBond M + -FB) on human gingival fibroblasts in terms of cytotoxicity, morphology and function. After in vitro exposure for up to 48 h, fibroblast viability was determined by the MTT assay determined, morphology by phase-contrast microscopy and migration by the scratch wound assay. Expression levels of IL1β, IL6, IL8, IL10, TNFα and VEGF genes were assessed by RT-PCR and their protein production by Western blot analysis. Apoptosis and cell cycle were analyzed by flow cytometry. OB and UB induced early morphological changes on fibroblasts (3 h) with extended cell death at 24 h/48 h. Gene expression of collagen type I and fibronectin increased fivefold compared with controls, elastin disappeared and elastase increased threefold, indicating gingival tissue tended to become fibrotic. Only UB and OB increased gene expression of inflammatory markers: IL1β at 3 and 48 h (up to about three times), IL6 and IL8 at 3 h (up to almost four times) which corresponded to the increase of the activated form NF-kB. All adhesives showed an effect on the functionality of fibroblasts with cytotoxic effect time and concentration dependent. Among all the OB and UB adhesives, they showed the greatest cell damage. The in-depth analysis of the effects of universal adhesives and possible functional effects represents an important information for the clinician towards choosing the most suitable adhesive system.

Objective: To compare the mechanical and biological features of a polymethylmethacrylate (PMMA) disc for CAD/CAM prostheses (test samples, TG) with a traditional resin (control samples, CG)., Methods: Mechanical analysis was performed using Dynamic Mechanical Analysis (DMA) and Brillouin's micro-spectroscopy. Human keratinocyte morphology and adhesion were analyzed by scanning electronic microscopy (SEM), cytotoxicity by the MTT assay, apoptosis by flow cytometry and p53, p21 and bcl2 gene expression by real time PCR., Results: TG exhibited a higher elastic modulus than CG (range 5100-5500 ± 114.3 MPa vs 3000-3300 ± 99.97 MPa). The Brillouin frequency was found at ω B = (15.50 ± 0.05) GHz for TG and at ω B&#95;1 = (15.50 ± 0.05) GHz and ω B&#95;2 = (15.0 ± 0.1) GHz for CG where two peaks were always present independently of the sample point. SEM analysis revealed that keratinocytes on TG disks appeared to be flattened with lamellipodia. Keratinocytes on CG disks rose above the substrate with cytoplasmatic filaments. MTT viability data at 3 h and 24 h showed TG was significantly less cytotoxic than CG (p < 0.001). No significant differences emerged in apoptosis on CG and TG. Real-time PCR showed p53 expression increased after 3 h by about 9-fold in keratinocytes on TG (p < 0.001) and about 5-fold in those on CG (p < 0.001). High p53 expression persisted after 24 h on both disks. No significant variations were observed in p21 and bcl2 expression at any time-point., Significance: PMMA resins, as used in CAD/CAM technology, displayed suitable biocompatible and mechanical properties for removable prostheses., (Copyright © 2020 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

A systematic review was performed to evaluate the biological effects of Cannabidiol (CBD), one of the major components of Cannabis Sativa, on normal human healthy cell populations in terms of cell viability, proliferation, migration, apoptosis and inflammation. Inclusion criteria were: studies on cell lines and primary cell culture from healthy donors, CBD exposure as variable, no CBD exposure as control and published in English language. Quality assessment was based on ToxR tool, with a score of reliability ranging from 15 to 18.Following the PRISMA statement, three independent reviewers performed both a manual and an electronic search using MEDLINE via PubMed, Scopus, Web of Science and Cochrane. From a total of 9437eligible articles, 29 studies have been selected. The average quality assessment score was 16.48.Theresults showed heterogeneous CBD concentration exposure (0.01-50 μM or 0.1 nmol/mL-15 mg/mL). The definition of a threshold limit would allow the identification of specific effects on expected outcomes. From the data obtained CBD resulted to inhibit cell viability in a dose-dependent manner above 2 μM, while in oral cell populations the inhibitory concentration is higher than 10 μM. Moreover, it was observed a significantly inhibition of cell migration and proliferation. On the contrary, it was highlighted a stimulation of apoptosis only at high doses (from 10 μM).Finally, CBD produced an anti-inflammatory effect, with a reduction of the pro-inflammatory cytokine gene expression and secretion. CBD down-regulated ROS production, although at high concentrations (16 μM) increased ROS-related genes expression. The diffusion of CBD for therapeutic and recreational uses require a precise definition of its potential biological effects. A thorough knowledge of these aspects would allow a safe use of this substance without any possible side effects., (Copyright © 2020. Published by Elsevier Masson SAS.)

Objective: To assess the effectiveness of lasers (at sub-ablative parameters) in reducing caries incidence compared with traditional prophylactic interventions (TPIs) when used alone or together with other TPIs such as pits and fissures sealant or fluoride gels or varnishes., Design: A systematic review. Data sources include Medline (via PubMed), Embase, Web of Science and the Cochrane Library (December 2019)., Eligibility Criteria: Only randomised trials (RCTs) and controlled clinical trials (CCTs) dealing with prophylactic lasers use (vs TPI or untreated teeth) were considered as eligible. We excluded in vitro and ex vivo studies., Data Extraction: Eligible studies were selected and data extracted independently by two reviewers. Risk of bias was assessed adopting the Cochrane Risk of Bias tool. Data on caries incidence, sealant retention, fluoride uptake, adverse events, treatment duration, patients' discomfort and cost-effectiveness ratio was extracted., Data Analysis: Extracted data were presented narratively due to the heterogeneity of included studies., Results: Seven RCTs and two CCTs, all with an evident risk of bias, met inclusion criteria, pooling together 269 individuals and 1628 teeth. CO 2 , neodymium-doped yttrium aluminium garnet, erbium-doped yttrium aluminum garnet (Er:YAG), erbium, chromium: yttrium scandium gallium garnet (Er,Cr:YSGG) and Argon lasers were used. In the permanent dentition, lasers only when used in combination with TPIs were effective in reducing caries when compared with untreated teeth (risk ratio (RR)=0.44 (0.20-0.97); Er:YAG laser) or with TPIs used alone (RR=0.39 (0.22-0.71); CO 2 laser). Moreover, Argon laser significantly improved the fluoride uptake into the enamel surfaces (ANalysis Of VAriance (ANOVA) tests: 95%, p<0.0001). Likewise, sealant retention improved when acid etching was performed on previously irradiated enamel fissures by CO 2 laser (RR=0.63 (0.38-1.04)) or Er:YAG laser (RR=0.54 (95% CI: 0.34 to 0.87)). In addition, laser resulted safe and well tolerated by patients., Conclusion: Despite some positive indications, an inadequate level of evidence was found in the included studies concerning the lasers' effectiveness in preventing caries. Further studies with a higher methodological quality level are required., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Aim: The aim of this review is to quantify the prevalence and type of malocclusion among children and adolescents during the different stages of dentition worldwide., Materials and Methods: Recent studies (from 2009 to 2019), published in Medline, Web of Science and Embase and orthodontic text-books have been comprehensively reviewed herein. The methodological quality of the included studies was assessed using STROBE criteria., Results: After screening 450 records and analysing 284 relevant full-text publications, 77 studies were included in this review. A good degree of evidence was obtained due to the medium-high methodological quality level of included studies. The worldwide prevalence of malocclusion was 56% (95% CI: 11-99), without differences in gender. The highest prevalence was in Africa (81%) and Europe (72%), followed by America (53%) and Asia (48%). The malocclusion prevalence score did not change from primary to permanent dentition with a common score of 54%. Malocclusion traits such as Angle's classes, overjet, overbite, and asymmetrical midline shift essentially did not change their prevalence during different dentitions. Conversely, traits such as cross-bite and diastema reduced their prevalence during permanent dentition, while scissor-bite and dental crowding increased their scores., Conclusion: The worldwide high prevalence of malocclusion and its early onset during childhood should induce policymakers as well as paediatric physicians and dentists to devise policies and adopt clinical strategies for preventing malocclusion since younger children's ages.

Pending the availability of vaccines to contain the SARS-CoV-2 pandemic, the current solution is "social distancing" with a reduction of dental treatments to those assessed as urgent and emergency cases. These treatments also involve Early Childhood Caries (ECC) due to the fact that this disease affects preschool children (a vulnerable population) and, in addition, shows a propensity to evolve into more serious complications (dental pain, infections). A narrative review was carried out to support a protocol for treating ECC with efficacious and safe (in terms of SARS-CoV-2 transmission) procedures. Protocol involves criteria for patients' selection remotely (telemedicine), and well-detailed criteria/equipment and hygiene procedures to combat against SARS-CoV-2 transmission. Moreover, the protocol proposes innovative caries treatments, named Minimally Invasive Treatments (MITs), well known in pedodontics for their high level of children's acceptance during dental care. MITs allow for caries removal (particularly in primary teeth) without any high-speed rotating instrument cooled with nebulized air-water spray (with high risk of virus environmental diffusion), usually adopted during traditional treatments. For evaluating MITs effectiveness in caries management, only Systematic Review and Randomized Controlled Trials (RCTs) were included in our study, without any risk of bias assessment. The indications proposed in this protocol could support clinicians for the temporary management of ECC until the SARS-CoV-2 pandemic ends.

Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [All the authors are employees of the GSK companies. F.B. is owner of patents on related topics].

Purpose: Critical care units, such as heart failure units, house inpatients with a compromised general health status that requires rigorous prevention of further complications. Oral health infections that gain access through the bloodstream or airway might represent such potential complications (eg, endocarditis pneumonia). Avoiding these critical occurrences requires that adequate oral health care be provided by nursing personnel. Here we assessed the knowledge of oral health care practices by nurses working in three Italian heart failure units in Umbria, Italy., Design: This was a cross-sectional study., Methods: Forty-four nurses were interviewed using a six-item modified Adams' questionnaire on the topic of oral health care. A multidisciplinary panel of experts established the criteria for answer correctness based on the most relevant dentistry literature evidence and judged each reply. The expected percentage of correctly replying nurses was 75%, and significant differences from this expected probability were calculated with one-sided binomial probability tests. Cronbach's α method was used to establish the questionnaire's internal consistency (reliability)., Results: For five out of six questionnaire items, the percentage of nurses who correctly answered was significantly lower than the expected value of probability. Lack of knowledge was found for usefulness of checking the patients' mouths (p=0.003), the most relevant lesions affecting the mouth (p=0.0001), the tools/solutions for cleaning the mouth and dentures (p= 0.0416), and drugs that affect the mouth and their side effects (p<0.0001)., Conclusion: In this study, few nurses working in heart failure units showed both an adequate willingness to check inpatients and a good knowledge of oral health care (significantly lower than the expected 75%). Further studies that use validated questionnaires and include more participants should be conducted to confirm and elaborate on our preliminary data., Competing Interests: The authors indicate no conflicts of interest in this scientific work., (© 2020 Cianetti et al.)

Aim: Caries lesions begin with enamel acid demineralisation mediated by microorganisms. Lasers with sub-ablative energy might act as a prophylactic intervention to reinforce enamel against lesions caused by acid., Materials and Methods: A systematic review of the literature was performed evaluating only in vitro studies published from 2010 to 2018. The research was performed using the following databases: Medline, Embase and the Web Of Science. A further search was performed consulting the list of references of the included studies as well as book chapters which dealt with this topic., Results: A total of 347 records were retrieved and, after their evaluation, 36 studies were included. CO2 lasers were the most described and effective device in preventing acid demineralisation. This type of laser was unique in improving the already positive results obtained with fluoride-based interventions. Er,Cr:YSGG (with fluencies > 8.5J/cm2), diode and argon lasers also improved enamel acid resistance (p-values ranging from 0.05 to 0.001) producing similar effects with fluoride-based interventions. Regarding the sealant retention outcome, the Er:YAG laser was able to perform an enamel etching which was as effective as the traditional acid etching with the advantage of being easier and usually well accepted by low-compliant patients (i.e. younger children). Nd:YAG presented the worst results. The most common structural changes after the laser irradiation were water and carbonate reduction in the enamel combined with a phosphate and calcium enamel content increase. Moreover, the calcium/phosphate ratio was found to reach the 1.67 ideal ratio., Conclusion: The in vitro studies that examined the prophylactic use of lasers for increasing enamel acid resistance presented interesting results that are enough to support a further in vivo experiment. This would entail the use of a clinical laser as an alternative or in combination with fluoride-based interventions.

Universal adhesives are the most important innovation in restorative dentistry. They are composed of different monomers, solvents and fillers. The potential cytotoxic effect of these materials is an important scientific aspect in recent literature. The aim of this study was to determine, using different in vitro techniques, the cytotoxicity evaluation of seven universal enamel-dental adhesives on human gingival fibroblasts. For this purpose, seven universal dental enamel adhesives have been evaluated by in vitro cytotoxicity tests using direct contact tests (an unpolymerized and a polymerized method) and an indirect contact test: preparation of extracts. The polymerized method showed a cytotoxicity range from 36% (G-PremioBond, GPB) to 79% (FuturaBond M+, FB). With the unpolymerized direct methods the range was from 4% (Prime&Bond Active, PBA) to 40% (Ibond Universal, IB) for undiluted adhesives; generally passing to the major dilutions the test showed a strong inhibitory activity by all the adhesives. Whereas with the indirect method by diluting the extracts of all dental adhesives the cell viability increased. The data obtained from the work has shown a lower cytotoxic effect of Optibond Solo Plus (OB) and Adhesive Universal (AU) with more reliable results with the extracts technique. The choice of reliable in vitro cytotoxic technique could represent, in dental practice, an important aid for clinical procedures in the use of adhesive systems., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Aim: Recently, the application of restorative materials containing metacrilate monomers in the conservative and paediatric dentistry has focused on the possible negative effects due to the use of these composites. In particular the release of monomers from reconstructions as a result of an insufficient polymerisation, can spread along the mucosal and dental tissues with potential immunological ed cytotoxic effects. Regarding to the importance of this issue, the aim of this study is to provide a descriptive review of the literature on potential local and systemic interactions of metacrylic and acrylic monomers with the immune system, both in vitro and in vivo., Results: The most highly used monomers in composite materials applied in conservative dentistry include: 2-hydroessietil- methacrylate (HEMA), triethylene glycol-dimethacrylate (TEGDMA), bisphenol A glycidyl-methacrylate (BisGMA) and urethane- dimethacrylate (UDMA). Different investigations have been performed for better understanding of the potential side effects of metacrylic monomers on immune system cells. Different factors such as cell population, exposure time and parameters more strictly connected to these materials, such as molecular weight, chemical composition and mechanical characteristics, seem to be directly involved in these reactions.

The use of multivalent glycoconjugate vaccines has dramatically contributed to reduce the incidence of meningococcal infectious disease. The advanced structural characterization of polysaccharide conjugates leads to enhancements in the quality and control of the products. Here, we report a novel nuclear magnetic resonance (NMR) method to confirm the identity and structural conformity (e.g., O -acetyl content) of saccharide antigens that comprise a licensed tetravalent meningococcal serogroups A, C, W, and Y vaccine. For the first time, the NMR methodology is applied on a formulation (licensed vaccine) containing a large excess of excipient (i.e., sucrose) without analytical sample pretreatment. This work confirms the applicability of a rapid and easy NMR assay on a multivalent conjugate vaccine, which might be extended to other combination vaccines that are already licensed or in clinical development., Competing Interests: The authors declare the following competing financial interest(s): All the authors are employees of the GSK companies. Francesco Berti (corresponding author) is owner of patents on related topics. This study was sponsored by GSK Biologicals SA.

The quality of dental arch impression has a substantial role in the precision of the intervention. It is traditionally acquired with resins that solidify when in contact with the air. Compared to that method, digital impression gives great advantages and, together with three-dimensional (3D) digitization devices, allows a simplification of the digital impression process. The growing adoption of such systems by a large number of dental clinics determines the need for an in-depth evaluation of the accuracy and the precision of the different systems. The aim of this work is to define a methodology for the evaluation of the accuracy and precision of 3D intraoral and desktop scanning systems, by using volumetric and linear methods. The replica of a tooth was realized with zirconium; afterward, high-accuracy point clouds of the master model were acquired by a coordinate measurement machine (CMM). In this way, the dimensions of the replica were accurately known. An intraoral scanner (I) and three desktops (D1, D2, D3) were then used to scan the replica. The geometry resulting from the CMM was compared with the ones derived from the scanners, using two different commercial programs (Geomagic and 3-Matic) and a custom-developed algorithm (MATLAB). Geomagic showed the mean values to be in a range from 0.0286 mm (D1) to 0.1654 mm (I), while 3-Matic showed mean values from -0.0396 mm (D1) to 0.1303 mm (I). MATLAB results ranged from 0.00014 mm (D1) to 0.00049 mm (D2). The probability distributions of the volumetric error of the measurements obtained with the different scanners allow a direct comparison of their performances. For the results given by our study, the volumetric approach that we adopted appears to be an excellent system of analysis.

This clinical report describes the use of a 3-dimensional (3D) printer to create an individual mold for delivering high-dose-rate interventional radiotherapy for hard palate cancer. The maxillary teeth and palate were scanned with an intraoral scanner (3Shape TRIOS 3). The scan was transformed into a mesh using the standard tessellation language (STL) format and aligned with Digital Imaging and Communications in Medicine (DICOM) computed tomography (CT) images using free Blue Sky Plan 4 planning software. A mold was generated by tracing a guideline around the gingival margins of the maxillary teeth and palate on the scan mesh in accordance with established parameters. All data were imported into computer-aided design (CAD) software. For this patient, 3 parallel 2.2-mm-diameter ducts were placed 10 mm from each other in the mold mesh. A CT scan of the patient's mouth with the mold in place was used for treatment planning. Treatment was delivered by means of microSelectron digital afterloading., (Copyright © 2018 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

The study evaluated the effects of 4 wt% nanohydroxyapatite (HA), 6 wt% zinc l-carnosine (MDA) and 1.5 wt% Ciprofloxacin (AB) on the mechanical, thermal and biological properties of glass ionomer cements (GIC). Filler and additive concentrations were selected after a previous study had tested single components and different percentages. Specimens included five silicon molds of each GIC cement for all tests. They were stored at room temperature for 24 h from specimen collection to analysis. Mechanical tests, calorimetric analysis, morphological investigation, antibacterial and cell viability assays were conducted. One-way analysis of variance (ANOVA) was used for data analysis with significance set at p < 0.05. Adding HA, MDA and AB to GICs modified their thermal, mechanical and microbiological properties. Polymerization increased. A slight decrease in the compressive strength of modified GICs was observed in dry condition (p < 0.05). Cement extracts affected cell viability in relation to extract dilution. Mechanical behavior improved in modified glass ionomer cements, especially with the powder formulated antibiotic. Overall cytotoxicity was reduced. Therefore adding nanohydroxyapatite, antibiotic and a mucosal defensive agent to conventional glass ionomer cement in special need patients could improve the clinical, preventive and therapeutic performance of the cements, without altering their mechanical properties., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Adjuvants are necessary to enable vaccine development against a significant number of challenging pathogens for which effective vaccines are not available. We engineered a novel small-molecule immune potentiator, a benzonaphthyridine agonist targeting toll-like receptor 7 (TLR7), as a vaccine adjuvant. TLR7 agonist (TLR7a) was engineered to be adsorbed onto aluminum hydroxide (AlOH), and the resulting AlOH/TLR7a was evaluated as a vaccine adjuvant. AlOH/TLR7a exploits the flexibility of AlOH formulations, has an application in many vaccine candidates, and induced good efficacy and safety profiles against all tested antigens (bacterial- and viral-derived protein antigens, toxoids, glycoconjugates, and so forth) in many animal models, including nonhuman primates. In this article, we describe the outcome of the physicochemical characterization of AlOH/TLR7a. Reverse-phase ultra performance liquid chromatography, confocal microscopy, flow cytometry, zeta potential, and phosphophilicity assays were used as tools to demonstrate the association of TLR7a to AlOH and to characterize this novel formulation. Raman spectroscopy, nuclear magnetic resonance, and mass spectroscopy were also used to investigate the interaction between TLR7a and AlOH (data not shown). This pivotal work paved the way for AlOH/TLR7a to progress into the clinic for evaluation as an adjuvant platform for vaccines against challenging preventable diseases., (Copyright © 2018 American Pharmacists Association®. All rights reserved.)

Objectives: To evaluate the effectiveness and degree of acceptance by children and adolescents of the use of oscillating tips compared with rotating drills., Design: Systematic review., Data Sources: PubMed, the Cochrane Central Register of Controlled Trials (CENTRAL), the Cochrane Library and Web of Science (October 2017)., Eligibility Criteria: Controlled randomised or non-randomised trials that evaluated sonic and ultrasonic oscillating devices versus rotating drill., Data Extraction: Eligible studies were selected and data extracted independently by two reviewers. Risk of bias was assessed using the Cochrane Method., Results: Two controlled clinical trials comprising 123 children aged 2-12 years old were identified. Both trials were at high risk of selection bias and unclear risk of detection bias. In one trial, pain due to the use of oscillating drill resulted lower than employing rotating drill (Verbal Hochman Scale: RR 0.64 (95% CI 0.41 to 1.00); Visual Facial Expression Scale: RR 0.64 (95% CI 0.44 to 0.94)). In another study, compared with traditional drill ultrasonic tip was associated with a lower level of patient's discomfort (RR 0.40 (95% CI 0.20 to 0.79)) but not with dental anxiety (RR 1.29 (95% CI 0.97 to 1.71)). The effectiveness of the removal of caries as well as fillings durability were only considered in one study, but no statistically significant differences were found between the two interventions., Conclusions: The evidence based on two low-quality studies was insufficient to conclude that the use of oscillating tips for the management of pain and dental fear in children or adolescents compared with rotating drills was more effective., Competing Interests: Competing interests: None declared., (© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

Aim: Children's dental fear and anxiety (DFA) causes significant problems in clinical practice. The 15-item Children's Fear Survey Schedule - Dental Subscale (CFSS-DS) and the 8-item Modified Child Dental Anxiety Scale (MCDAS) are the most widely used measures of dental fear in children. The aim of this study is to examine the reliability and validity of the Italian versions of the CFSS-DS and MCDAS, also in comparison with a simple visual analogue scale (VAS)., Materials and Methods: The CFSS-DS and MCDAS were translated into Italian by a consensus panel of experts and administered to 210 dental patients aged 4-11 years from three Italian Institutions. Internal reliability was assessed using the Cronbach's alpha correlation. A sub-sample of 60 children was selected for test-retest analysis. CFSS-DS and MCDAS, plus a VAS scale, rated both by children and parents, were validated using as gold standard the 4-item Frankl scale for behaviours assessed by dentists., Results: Mean CFSS-DS score was 30.8 (SD: 11.1) and mean MCDAS score was 17.9 (SD: 7.2), significantly higher among children aged 4-7 years and among children at their first dental visit. The alpha value for internal reliability was 0.90 (95%, CI= 0.88-0.92) for CFSS-DS and 0.87 (95% CI=0.85-0.90) for MCDAS. Both CFSS-DS and MCDAS showed good test-retest reliability (rsp= 0.80; p<0.001 for both scales). CFSS-DS and MCDAS predicted a Frankl score &#8804;2 (i.e., indicating children with an uncooperative behaviour) with a fair accuracy (AUC=0.69 and AUC=0.68, respectively). The VAS scale was more effective in predicting a negative behaviour (AUC=0.78). The scales self-reported by children were only slightly more accurate than those reported by parents., Conclusion: The Italian versions of the CFSS-DS and MCDAS are valid and reliable tools for the assessment of dental fear in Italian children aged 4-11 years. A simple, one-item VAS, and dental fear and anxiety evaluation by parents may be valid and quick alternatives to multi-item indices to predict an uncooperative children behaviour.