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Abstract Novel technologies such as complex magnetic fields—CMFs represent an eco-sustainable proposal to counteract the infection associated to resistant microorganisms. The aim of this study was to evaluate the effect of two CMF programs (STRESS, ANTIBACTERIAL) against clinical antifungal resistant C. albicans also evaluating their uneffectiveness on gingival fibroblasts (hGFs). The STRESS program was more efficacious on C. albicans biofilm with up to 64.37% ± 10.80 of biomass and up to 99.19% ± 0.06 CFU/ml reductions in respect to the control also inducing an alteration of lipidic structure of the membrane. The MTT assay showed no CMFs negative effects on the viability of hGFs with a major ROS production with the ANTIBACTERIAL program at 3 and 24 h. For the wound healing assay, STRESS program showed the best effect in terms of the rate migration at 24 h, showing statistical significance of p

Objectives: This systematic review aimed to analyse the published evidence for the use of non-invasive methods for the early detection of oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMDs). Methods: The literature was systematically searched through several databases: PubMed, Cochrane Library, and Web of Science. Additional exploration was performed through cross-checks on the bibliographies of selected reviews. The inclusion criteria involved studies assessing the application of non-invasive tests on humans in the screening, diagnosis, or surveillance of OSCC or OPMDs and reporting sensitivity (SE) and specificity (SP). The Newcastle–Ottawa scale (NOS) was applied to assess the quality of the studies included. Results: The search strategy resulted in 8012 preliminary records. After a duplicate check, 116 titles remained. After abstract analysis, 70 papers remained. After full text analysis, only 54 of the 70 papers fit the inclusion criteria (28 were original articles and 26 were reviews). Those 26 reviews were used to manually search for further original articles. From this last search, 33 original articles were found. Thus, a total of 61 original studies were included and investigated. Findings from this systematic review indicate useful information, such as a description of the mechanisms, ease of use, limitations, and SE and SP values, to drive the choice of the optimal minimally invasive method to be utilized as an adjunctive tool to examine the suspicious lesions. Conclusions: Each of the analysed tools can be improved or implemented, considering their high SE and low SP. Despite advancements, incisional biopsy continues to be the gold standard for the definitive diagnosis of oral cancer and precancerous lesions. Further research and development are essential to improving the sensitivity, specificity, and reliability of non-invasive tools for widespread clinical application.

Abstract Background Spread worldwide through droplets, the Virus Sars-Cov-19 has caused a global health emergency alarm. In order to limit its spread, the use of masks has become part of the daily life of the entire population, however, little is known about its constant use and the changes generated in the oral cavity. This work aims to investigate correlations between the continuous use of masks covering the nose and mouth for 3 h and changes in the ecological factors of the oral cavity. Methods 34 volunteers were divided into 2 groups: wear only the filtering facepiece code 2 (FFP2) mask (Group A) and wear the FFP2 mask covered by a surgical mask (Group B). Measurement of Volatile Organic Compounds (VOCs), saliva rehydration and consistency test, collection of basal saliva and saliva stimulated with paraffin gum and mucosal swab were collected and analyzed at two times: before using the mask(s) (T0) and 3 h after continuous use of the mask(s) (T1). Results The results indicated a significant difference between the groups, in which the basal saliva volume and pH and the peaks of VOCs increased for group B between T0 and T1. The rehydration time decreased and the volume and pH of the stimulated saliva increased, but with no significant difference between the groups. Furthermore, group B showed a significant decrease in Candida albicans Colony Forming Units (CFUs) and Total Bacterial Count (TBC) between T0 and T1. Conclusion It is concluded that the prolonged use of the FFP2 mask covered by a surgical mask can generate oral alterations in the user.

The interaction between oral bacteria and dental implant surfaces is a critical factor in the success and longevity of dental implants. With advancements in additive manufacturing technologies, selective laser melting (SLM) has emerged as a prominent method for producing titanium implants with highly controlled microstructures and porosities. These 3D printed titanium surfaces offer significant benefits, such as enhanced osseointegration and improved mechanical properties. However, the same surface features that promote bone cell attachment and proliferation may also provide favorable conditions for bacterial adhesion and biofilm formation. Understanding the dynamics of these interactions is essential for developing implant surfaces that can effectively resist bacterial colonization while promoting tissue integration. This narrative review explores the complex interplay between oral bacteria and SLM-produced titanium porous surfaces, examining current research findings and potential strategies for optimizing implant design to mitigate the risks of infection and ensure successful clinical outcomes.

Oxidative stress and high levels of reactive oxygen species (ROS) are linked to various age-related diseases and chronic conditions, including damage to oral tissues. Dexamethasone (DEX), a widely used glucocorticoid in dentistry, can have side effects like increased ROS production and delayed wound healing. Resveratrol (RSV) is known for its antioxidant properties, but its limited bioavailability hinders its clinical use. This study investigated the potential of two RSV derivatives (1d and 1h) to address these limitations. The antioxidant abilities of 1d and 1h (5 μM) against DEX-induced oxidative stress (200 μM) were evaluated in human gingival fibroblasts (hGFs) and osteoblasts (hOBs). The effects of these compounds on cell viability, morphology, ROS levels, SOD activity, gene expression, and collagen production were evaluated. RSV derivatives, under DEX-induced oxidative stress condition, improved cell growth at 72 h (191.70 ± 10.92% for 1d+DEX and 184.80 ± 13.87% for 1h+DEX), morphology, and SOD activity (77.33 ± 3.35 OD for 1d+DEX; 76.87 ± 3.59 OD for 1h+DEX at 1 h), while reducing ROS levels (2417.33 ± 345.49 RFU for 1d+DEX and 1843.00 ± 98.53 RFU at 4 h), especially in hOBs. The co-treatment of RSV or derivatives with DEX restored the expression of genes that were downregulated by DEX, such as HO-1 (1.76 ± 0.05 for 1d+DEX and 1.79 ± 0.01 for 1h+DEX), CAT (0.97 ± 0.06 for 1d+DEX and 0.99 ± 0.03 for 1h+DEX), NRF2 (1.62 ± 0.04 for 1d+DEX and 1.91 ± 0.05 for 1h+DEX), SOD1 (1.63 ± 0.15 for 1d+DEX and 1.69 ± 0.04 for 1h+DEX). In addition, 1d and 1h preserved collagen production (111.79 ± 1.56 for 1d+DEX and 122.27 ± 1.56 for 1h+DEX). In conclusion, this study suggests that the RSV derivatives 1d and 1h hold promise as potential antioxidant agents to counteract DEX-induced oxidative stress. These findings contribute to the development of novel therapeutic strategies for managing oxidative stress-related oral conditions.

The imbalance in skin microbiota is characterized by an increased number of pathogens in respect to commensal microorganisms. Starting from a skin microbiota collection, the aim of this work was to evaluate the possible role of Pomegranate (Punica granatum L.) Peel Extract (PPE) in restoring the skin microbiota balance acting on Staphylococcus spp. PPE was extracted following green methodology by using n-butane and the Dimethyl Ether (DME) solvents and analyzed for phytochemical composition and antimicrobial activity. The PPE antimicrobial action was evaluated against Gram +, Gram - bacteria and yeast reference strains and the most effective extract was tested against the main skin microbiota isolated strains. PPE extracted with DME showed the best antimicrobial action with MICs ranging from 1 to 128 mg/mL; the main active compounds were Catechin, Quercetin, Vanillic acid and Gallic acid. The PPE in DME anti-adhesive effect was examined against S. epidermidis and S. aureus mono and dual-species biofilm formation by biomass quantification and CFU/mL determination. The extract toxicity was evaluated by using Galleria mellonella larvae in vivo model. The extract displayed a significant anti-adhesive activity with a remarkable species-specific action at 4 and 8 mg/mL against S. epidermidis and S. aureus mono and dual-species biofilms. PPE in DME could represent an eco-sustainable non-toxic strategy to affect the Staphylococcal skin colonization in a species-specific way. The innovation of this work is represented by the reuse of food waste to balance skin microbiota.

This study aimed to evaluate the ability of photodynamic therapy, based on the use of a gel containing 5% delta aminolaevulinic acid (ALAD) for 45′ followed by irradiation with 630 nm LED (PDT) for 7′, to eradicate Candida albicans strains without damaging the gingiva. C. albicans oral strains and gingival fibroblasts (hGFs) were used to achieve these goals. The potential antifungal effects on a clinical resistant C. albicans S5 strain were evaluated in terms of biofilm biomass, colony forming units (CFU/mL) count, cell viability by live/dead analysis, and fluidity membrane changes. Concerning the hGFs, viability assays, morphological analysis (optical, scanning electronic (SEM), and confocal laser scanning (CLSM) microscopes), and assays for reactive oxygen species (ROS) and collagen production were performed. ALAD-mediated aPDT (ALAD-aPDT) treatment showed significant anti-biofilm activity against C. albicans S5, as confirmed by a reduction in both the biofilm biomass and CFUs/mL. The cell viability was strongly affected by the treatment, while on the contrary, the fluidity of the membrane remained unchanged. The results for the hGFs showed an absence of cytotoxicity and no morphological differences in cells subjected to ALAD-aPDT expected for CLSM results that exhibited an increase in the thickening of actin filaments. ROS production was augmented only at 0 h and 3 h, while the collagen appeared enhanced 7 days after the treatment.

Microbial adhesion on dental restorative materials may jeopardize the restorative treatment long-term outcome. The goal of this in vitro study was to assess Candida albicans capability to adhere and form a biofilm on the surface of heat-cured dental composites having different formulations but subjected to identical surface treatments and polymerization protocols. Three commercially available composites were evaluated: GrandioSO (GR), Venus Diamond (VD) and Enamel Plus HRi Biofunction (BF). Cylindrical specimens were prepared for quantitative determination of C. albicans S5 planktonic CFU count, sessile cells CFU count and biomass optical density (OD570 nm). Qualitative Concanavalin-A assays (for extracellular polymeric substances of a biofilm matrix) and Scanning Electron Microscope (SEM) analyses (for the morphology of sessile colonies) were also performed. Focusing on planktonic CFU count, a slight but not significant reduction was observed with VD as compared to GR. Regarding sessile cells CFU count and biomass OD570 nm, a significant increase was observed for VD compared to GR and BF. Concanavalin-A assays and SEM analyses confirmed the quantitative results. Different formulations of commercially available resin composites may differently interact with C. albicans. The present results showed a relatively more pronounced antiadhesive effect for BF and GR, with a reduction in sessile cells CFU count and biomass quantification.

This work aimed to evaluate and compare the antimicrobial actions and effects over time of eight types of mouthwash, based on the impact of chlorhexidine on the main microorganisms that are responsible for oral diseases: Enterococcus faecalis, Pseudomonas aeruginosa, and Candida albicans. The mouthwashes’ antimicrobial action was determined in terms of their minimum inhibitory concentration (MIC), minimum bactericidal/fungicidal concentration (MBC/MFC), and time-kill curves at different contact times (10 s, 30 s, 60 s, 5 min, 15 min, 30 min, and 60 min), against selected oral microorganisms. All the mouthwashes showed a notable effect against C. albicans (MICs ranging from 0.02% to 0.09%), and higher MIC values were recorded with P. aeruginosa (1.56% to >50%). In general, the mouthwashes showed similar antimicrobial effects at reduced contact times (10, 30, and 60 s) against all the tested microorganisms, except with P. aeruginosa, for which the most significant effect was observed with a long time (15, 30, and 60 min). The results demonstrate significant differences in the antimicrobial actions of the tested mouthwashes, although all contained chlorhexidine and most of them also contained cetylpyridinium chloride. The relevant antimicrobial effects of all the tested mouthwashes, and those with the best higher antimicrobial action, were recorded by A—GUM® PAROEX®A and B—GUM® PAROEX®, considering their effects against the resistant microorganisms and their MIC values.

Helicobacter pylori colonizes human gastric mucosa, overcoming stressful conditions and entering in a dormant state. This study evaluated: (i) H. pylori’s physiological changes from active to viable-but-non-culturable (VBNC) and persister (AP) states, establishing times/conditions; (ii) the ability of vitamin C to interfere with dormancy generation/resuscitation. A dormant state was induced in clinical MDR H. pylori 10A/13 by: nutrient starvation (for VBNC generation), incubating in an unenriched medium (Brucella broth) or saline solution (SS), and (for AP generation) treatment with 10xMIC amoxicillin (AMX). The samples were monitored after 24, 48, and 72 h, 8–14 days by OD600, CFUs/mL, Live/Dead staining, and an MTT viability test. Afterwards, vitamin C was added to the H. pylori suspension before/after the generation of dormant states, and monitoring took place at 24, 48, and 72 h. The VBNC state was generated after 8 days in SS, and the AP state in AMX for 48 h. Vitamin C reduced its entry into a VBNC state. In AP cells, Vitamin C delayed entry, decreasing viable coccal cells and increasing bacillary/U-shaped bacteria. Vitamin C increased resuscitation (60%) in the VBNC state and reduced the aggregates of the AP state. Vitamin C reduced the incidence of dormant states, promoting the resuscitation rate. Pretreatment with Vitamin C could favor the selection of microbial vegetative forms that are more susceptible to H. pylori therapeutical schemes.

Guided bone regeneration (GBR) comprehends the application of membranes to drive bone healing and to exclude non-osteogenic tissues from interfering with bone regeneration. However, the membranes may be exposed to bacterial attack, with the risk of failure of the GBR. Recently, an antibacterial photodynamic protocol (ALAD-PDT) based on a gel with 5% 5-aminolevulinic acid incubated for 45 min and irradiated for 7 min by a LED light at 630 nm, also showed a pro-proliferative effect on human fibroblasts and osteoblasts. The present study hypothesized that the functionalization of a porcine cortical membrane (soft-curved lamina, OsteoBiol) with ALAD-PDT might promote its osteoconductive properties. TEST 1 aimed to verify the response of osteoblasts seeded on lamina with respect to the plate surface (CTRL). TEST 2 aimed to investigate the effects of ALAD-PDT on the osteoblasts cultured on the lamina. SEM analyses were performed to study the topographical characteristics of the membrane surface, the adhesion, and the morphology of cells at 3 days. The viability was assessed at 3 days, the ALP activity at 7 days, and calcium deposition at 14 days. Results showed the porous surface of the lamina and the increase in cell attachment of osteoblasts with respect to controls. The proliferation, the ALP, and bone mineralization activity of osteoblasts seeded on lamina resulted in being significantly higher (p < 0.0001) than controls. Results also showed an additional significative enhancement (p < 0.0001) in the proliferative rate in ALP and calcium deposition after applying ALAD-PDT. In conclusion, the functionalization of the cortical membranes cultured with osteoblasts with the ALAD-PDT improved their osteoconductive properties.

Background: In this ex vivo study, the aim was to evaluate the effects of ALAD and red light on Enterococcus faecalis in infected root canals using a special intracanal fiber. Methods: A total of 70 extracted, single-rooted teeth were used. The teeth were decoronated at the length of the roots to approximately 15 mm and then instrumented. The apical foramen was sealed by composite resin, and the root canals were infected with a pure culture of E. faecalis ATCC 29212 for eight days at 37 °C. Following the contamination period, the roots were divided into seven groups, including the positive and negative control groups, and treated as follows: ALAD 45 min; red light activation 7 min; ALAD 45 min and red-light activation 7 min; sodium hypochlorite 2.5% 15 min; sodium hypochlorite 1% 15 min. The samples were taken by three sterile paper points, transferred to tubes containing 1 mL of PBS, and immediately processed for the number of colony-forming units and the cell viability by using live/dead. Results: The best treatment is obtained with 2.5% NaOCl. Except for ALAD + red light vs. 1% NaOCl, a statistically significant difference is recorded for all treatments. The combination of 2.5% NaOCl and ALAD + 7 min irradiation produces an evident killing effect on the E. faecalis cells. On the other hand, 1% NaOCl is ineffective for the viability action, with 25% of dead cells stained in red. Conclusions: This ex vivo study shows that ALAD gel with light irradiation is an efficacious protocol that exerts a potent antibacterial activity against E. faecalis in infected root canals.

Recently, there has been an increasing interest in finding new approaches to manage oral wound healing. Although resveratrol (RSV) exhibited many biological properties, such as antioxidant and anti-inflammatory activities, its use as a drug is limited by unfavorable bioavailability. This study aimed to investigate a series of RSV derivatives (1a–j) with better pharmacokinetic profiles. At first, their cytocompatibility at different concentrations was tested on gingival fibroblasts (HGFs). Among them, derivatives 1d and 1h significantly increased cell viability compared to the reference compound RSV. Thus, 1d and 1h were investigated for cytotoxicity, proliferation, and gene expression in HGFs, endothelial cells (HUVECs), and oral osteoblasts (HOBs), which are the main cells involved in oral wound healing. For HUVECs and HGFs, the morphology was also evaluated, while for HOBs ALP and mineralization were observed. The results showed that both 1d and 1h did not exert negative effects on cell viability, and at a lower concentration (5 µM) both even significantly enhanced the proliferative rate, compared to RSV. The morphology observations pointed out that the density of HUVECs and HGFs was promoted by 1d and 1h (5 µM) and mineralization was promoted in HOBs. Moreover, 1d and 1h (5 µM) induced a higher eNOS mRNA level in HUVECs, higher COL1 mRNA in HGFs, and higher OCN in HOBs, compared to RSV. The appreciable physicochemical properties and good enzymatic and chemical stability of 1d and 1h, along with their promising biological properties, provide the scientific basis for further studies leading to the development of RSV-based agents useful in oral tissue repair.

This in vitro study aims to evaluate the bacterial microleakage of three conometric connections. Sixty dental implants (3P implafavourite Scalenghe) were divided in groups (n = 20): Cone–Morse with passing screw (Group 1); Cone–Morse with solid abutment (Group 2); and Conometric connection with esthetic abutment (Group 3). The implants were fixed in resin bases. Then, 1.0 µL of Streptococcus oralis (SO) was inoculated in the internal platform in 10 fixtures for each group, and another 10 were inoculated with Pseudomonas aeruginosa (PA). The abutments were then screwed, and five implants from each subgroup were randomly selected for SEM inspection to ensure that the abutments were installed correctly. Data were submitted to statistical analysis, ANOVA and Fisher’s Least Significant Difference (p ≤ 0.05). The turbidity of the broth was monitored for 14 days of follow-up in order to determine the penetration of the bacterial suspension into the surrounding solution, but the observation of the samples lasted until the 90th day, in which there was no difference between the two. Microbial contamination was found in 30%, 20%, and 50% of Group 1, Group 2, and Group 3, respectively, but there were no statistically significant differences between the groups, and PA showed greater infiltration than SO. Although no statistically significant differences were found, cone morse connections showed lower infiltration percentages, respective to the conometric connection with 18° angle.

Resistant wound microorganisms are becoming an extremely serious challenge in the process of treating infected chronic wounds, leading to impaired healing. Thus, additional approaches should be taken into consideration to improve the healing process. The use of natural extracts can represent a valid alternative to treat/control the microbial infections in wounds. This study investigates the antimicrobial/antivirulence effects of Capparis spinose aqueous extract against the main chronic wound pathogens: Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. The extract shows phenolic characterization with rutin (1.8 ± 0.14 μg/mg) as the major compound and antibacterial effect against bacteria (S. aureus PECHA 10 MIC 6.25%; P. aeruginosa PECHA 4 MIC 12.50%) without action against C. albicans (MIC and MFC ≥ 50%). Capparis spinose also shows a significant antivirulence effect in terms of antimotility/antibiofilm actions. In particular, the extract acts (i) on P. aeruginosa both increasing its swimming and swarming motility favoring the planktonic phenotype and reducing its adhesive capability, (ii) on S. aureus and P. aeruginosa biofilm formation reducing both the biomass and CFU/ml. Furthermore, the extract significantly displays the reduction of a dual-species S. aureus and P. aeruginosa Lubbock chronic wound biofilm, a complex model that mimics the realistic in vivo microbial spatial distribution in wounds. The results suggest that C. spinose aqueous extract could represent an innovative eco-friendly strategy to prevent/control the wound microbial infection.

Abstract The aim of this study was to evaluate the interaction between Streptococcus oralis and Polyetheretherketone (PEEK), a novel material recently introduced in implantology. The topographical characterization and the Streptococcus oralis adhesion on this material were compared with other titanium surfaces, currently used for the production of dental implants: machined and double etched (DAE). The superficial micro-roughness of the PEEK discs was analyzed by scanning electron microscopy (SEM) and, the Energy Dispersive Spectrometer (EDS) analyzed their chemical composition. Atomic Force Microscopy (AFM) was used to characterize the micro-topography and the sessile method to evaluate the wettability of the samples. Microbiological analysis measured the colony forming units (CFUs), the biomass (OD570 detection) and the cell viability after 24 and 48 h after Streptococcus oralis cultivation on the different discs, that were previously incubated with saliva. Results showed that PEEK was characterized by a micro-roughness that was similar to machined titanium but at nano-level the nano-roughness was significantly higher in respect to the other samples. The EDS showed that PEEK superficial composition was characterized mainly by Carbonium and Oxygen. The hydrophilicity and wetting properties of PEEK were similar to machined titanium; on the contrary, double etched discs (DAE) samples were characterized by significantly higher levels (p

Chronic wounds have harmful effects on both patients and healthcare systems. Wound chronicity is attributed to an impaired healing process due to several host and local factors that affect healing pathways. The resulting ulcers contain a wide variety of microorganisms that are mostly resistant to antimicrobials and possess the ability to form mono/poly-microbial biofilms. The search for new, effective and safe compounds to handle chronic wounds has come a long way throughout the history of medicine, which has included several studies and trials of conventional treatments. Treatments focus on fighting the microbial colonization that develops in the wound by multidrug resistant pathogens. The development of molecular medicine, especially in antibacterial agents, needs an in vitro model similar to the in vivo chronic wound environment to evaluate the efficacy of antimicrobial agents. The Lubbock chronic wound biofilm (LCWB) model is an in vitro model developed to mimic the pathogen colonization and the biofilm formation of a real chronic wound, and it is suitable to screen the antibacterial activity of innovative compounds. In this review, we focused on the characteristics of chronic wound biofilms and the contribution of the LCWB model both to the study of wound poly-microbial biofilms and as a model for novel treatment strategies.

The present study aims to discuss the main factors involving the use of 5-aminolevulinic acid together with red LED light and its application in endodontic treatment through a narrative review and a case report. Persistence of microorganisms remaining on chemical-mechanical preparation or intracanal dressing is reported as the leading cause of failure in endodontics. Photodynamic therapy has become a promising antimicrobial strategy as an aid to endodontic treatment. Being easy and quick to apply, it can be used both in a single session and in several sessions, as well as not allowing forms of microbial resistance. 5-aminolevulinic acid in combination with red LED light has recently been studied in many branches of medicine, with good results against numerous types of bacteria including Enterococuss faecalis. The case report showed how bacterial count of CFU decreased by half (210 CFU/mL), after 45 min of irrigation with a gel containing 5% of 5-aminolevulinic acid compared to the sample before irrigation (420 CFU/mL). The subsequent irradiation of red LED light for 7 min, the bacterial count was equal to 0. Thus, it is concluded that the use of 5-aminolevulinic acid together with red LED light is effective in endodontic treatment.

The objective of this study is to evaluate, in vitro, the microleakage of bacteria of 3 different implant connections for a period of 14 days. 60 dental implants (AoN) (n = 20) were distinguished into three groups, accordingly to the type of connection: External Hexagon (EH), Internal Hexagon (IH), and Cone Morse (CM) connection. All implants were inserted and fixed on sterile special vinyl support. Ten fixtures for each group were inoculated in the internal platform with 1.0 μL of Streptococcus oralis (SO) and the other ten with the same amount of Pseudomonas aeruginosa (PA). The penetration of bacterial suspension into the surrounding solution was determined by the observation of the turbidity of the broth. Five implants for each sub-group were randomly observed at SEM, to verify the correct fitting of the abutments. Considering the total of the samples analyzed, CM showed significantly lower bacterial contamination, with respect to IH. In particular, bacterial contamination was found in 45%, 55%, and 20% of EH, IH, and CM, respectively. Analyzing results for the type of inoculated bacteria, P. aeruginosa showed a higher ability to contaminate all the connections, with respect to S. oralis.

Innovative non-antibiotic compounds such as graphene oxide (GO) and light-emitting diodes (LEDs) may represent a valid strategy for managing chronic wound infections related to resistant pathogens. This study aimed to evaluate 630 nm LED and 880 nm LED ability to enhance the GO antimicrobial activity against Staphylococcus aureus- and Pseudomonas aeruginosa-resistant strains in a dual-species biofilm in the Lubbock chronic wound biofilm (LCWB) model. The effect of a 630 nm LED, alone or plus 5-aminolevulinic acid (ALAD)-mediated photodynamic therapy (PDT) (ALAD-PDT), or an 880 nm LED on the GO (50 mg/l) action was evaluated by determining the CFU/mg reductions, live/dead analysis, scanning electron microscope observation, and reactive oxygen species assay. Among the LCWBs, the best effect was obtained with GO irradiated with ALAD-PDT, with percentages of CFU/mg reduction up to 78.96% ± 0.21 and 95.17% ± 2.56 for S. aureus and P. aeruginosa, respectively. The microscope images showed a reduction in the cell number and viability when treated with GO + ALAD-PDT. In addition, increased ROS production was detected. No differences were recorded when GO was irradiated with an 880 nm LED versus GO alone. The obtained results suggest that treatment with GO irradiated with ALAD-PDT represents a valid, sustainable strategy to counteract the polymicrobial colonization of chronic wounds.

(1) Background: Recently, novel dental implants that are characterized by different levels of surface roughness in the distinct parts of the fixture’s body have been introduced in the market. These surface characteristics could affect the primary stability of the implants. The aim of this in vitro study was to compare the primary stability of short and long implants, characterized by multiscale surface roughness, inserted on polyurethane blocks. The secondary aim was to understand if the implant length could be a crucial factor in the decision-making in immediate or rather than delayed loading protocol in the different bone densities. (2) Methods: A total of 20 cylindrical dental implants with a diameter of 5.0 mm were tested for the lengths 6.0 mm (short implants) versus 13.0 mm (long implants) on two different solid rigid polyurethane blocks (20 and 30 PCF). The primary stability was evaluated by measuring the insertion torque value (ITV), the removal torque (RTV), and the resonance frequency analysis RFA. (3) Results: The values of ITV, RTV, and RFA showed the same trend in all measurements. Long implants showed a significantly higher primary stability on 30 PCF blocks that present mechanical properties similar to high-density bone. On the contrary, no relevant differences were found on 20 PCF blocks, which mimic trabecular bone density. (4) Conclusions: The impact of fixture length on the primary stability of implants with multiscale surface roughness is significant in 30 PCF polyurethane corresponding to higher bone density, but not in lower ones.

Antibacterial and antibiofilm properties of restorative dental materials may improve restorative treatment outcomes. The aim of this in vitro study was to evaluate Streptococcus mutans capability to adhere and form biofilm on the surface of three commercially available composite resins (CRs) with different chemical compositions: GrandioSO (VOCO), Venus Diamond (VD), and Clearfil Majesty (ES-2). Disk-shaped specimens were manufactured by light-curing the CRs through two glass slides to maintain a perfectly standardized surface topography. Specimens were subjected to Planktonic OD600nm, Planktonic CFU count, Planktonic MTT, Planktonic live/dead, Adherent Bacteria CFU count, Biomass Quantification OD570nm, Adherent Bacteria MTT, Concanavalin A, and Scanning Electron Microscope analysis. In presence of VOCO, VD, and ES2, both Planktonic CFU count and Planktonic OD600nm were significantly reduced compared to that of control. The amount of Adherent CFUs, biofilm Biomass, metabolic activity, and extracellular polymeric substances were significantly reduced in VOCO, compared to those of ES2 and VD. Results demonstrated that in presence of the same surface properties, chemical composition might significantly influence the in vitro bacterial adhesion/proliferation on resin composites. Additional studies seem necessary to confirm the present results.

The use of a new gel containing aminolevulinic acid and red light (ALAD–PDI) was tested in order to counteract bacterial biofilm growth on different titanium implant surfaces. The varying antibacterial efficacy of ALAD–PDI against biofilm growth on several titanium surfaces was also evaluated. A total of 60 titanium discs (30 machined and 30 double-acid etched, DAE) were pre-incubated with saliva and then incubated for 24 h with Streptococcus oralis to form bacterial biofilm. Four different groups were distinguished: two exposed groups (MACHINED and DAE discs), covered with S. oralis biofilm and subjected to ALAD + PDI, and two unexposed groups, with the same surfaces and bacteria, but without the ALAD + PDI (positive controls). Negative controls were non-inoculated discs alone and combined with the gel (ALAD) without the broth cultures. After a further 24 h of anaerobic incubation, all groups were evaluated for colony-forming units (CFUs) and biofilm biomass, imaged via scanning electron microscope, and tested for cell viability via LIVE/DEAD analysis. CFUs and biofilm biomass had significantly higher presence on unexposed samples. ALAD–PDI significantly decreased the number of bacterial CFUs on both exposed surfaces, but without any statistically significant differences among them. Live/dead staining showed the presence of 100% red dead cells on both exposed samples, unlike in unexposed groups. Treatment with ALAD + red light is an effective protocol to counteract the S. oralis biofilm deposited on titanium surfaces with different tomography.

The topography of implant surfaces influences the interaction relationship between material and bacteria. The aim of this work was to characterize a novel 3D titanium surface, produced using Selective Laser Sintering (SLS), and to compare the bacterial interaction with machined and double acid etching (DAE) discs. The surface was characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), and Energy Dispersive X-ray Spectrometry (EDX). The wettability was measured using the sessile method. The microbiological investigation consisted in the cultivation of a bacterial pioneer, Streptococcus oralis, on titanium surfaces, previously covered by human saliva in order to form the acquired pellicle. Then, colony forming units (CFUs), biofilm biomass quantification, analyses of viable and dead cells, and SEM observation were determined after 24 h of S. oralis biofilm formation on the different discs. A significantly higher nano-roughness with respect to the other two groups characterized the novel 3D surface, but the wettability was similar to that of machined samples. The microbiological assays demonstrated that the 3D discs reported significantly lower values of CFUs and biofilm biomass with respect to machined surfaces; however, no significant differences were found with the DAE surfaces. The live/dead staining confirmed the lower percentage of living cells on DAE and 3D surfaces compared with the machined. This novel 3D surface produced by SLS presented a high antiadhesive and antibiofilm activity.

The "Quality of Life" project arises from the need to observe and define the existing interconnection between a "healthy" lifestyle and the quality of life perceived by people. Starting from the W.H.O. (World Health Organization) definitions of quality of life and health, and analyzing the studies that describe the benefits of both movement physical and mental, a protocol three months has been set for the pursuit of a diet and personalized training. For this purpose, 20 healthy subjects from the 1st year of the Physiotherapy degree, who had never practiced sports at an agonistic level, have been recruited. All individuals were subject to initial evaluations to define psycho-physical wellbeing, then they have been divided into two groups: the first group, defined as experimental, followed the protocol for the duration of the study, while the second, was only subject to evaluation. At the end of the three months period, the subjects of both groups were evaluated twice, the first immediately after the last training session, the second was evaluated one month from the latter. The final evaluation aimed at verifying a possible conservative effect on modification of experimental group parameters, even when physical activity and nutrition were not supervised by us. The ultimate goal of the project was to specifically analyze, after three months, the changes in cognitive memory capacity and concentration, the stress experienced in work or study, and psycho-physical wellbeing perceived by the concerned subjects. Data collected during the three evaluations showed that, in the experimental group, as our scientific protocol has altered all these parameters in terms of quality of life in positive terms, modifications are susceptible to changes in the event that physical activity and proper nutrition are not followed.

This study evaluates the effects of different programs of complex electromagnetic fields (C.M.F.s) on Candida albicans, in planktonic and sessile phase and on human gingival fibroblasts (HGF cells). In vitro cultures of C. albicans ATCC 10231 and HGF cells were exposed to different cycles of C.M.F.s defined as: oxidative stress, oxidative stress/antibacterial, antibacterial, antibacterial/oxidative stress. Colony forming units (CFUs), metabolic activity, cells viability (live/dead), cell morphology, filamentation analysis, and cytotoxicity assay were performed. The broth cultures, exposed to the different C.M.F.s, were grown on titanium discs for 48 h. The quantity comparisons of adhered C. albicans on surfaces were determined by CFUs and scanning electron microscopy. The C. albicans growth could be readily controlled with C.M.F.s reducing the number of cultivable planktonic cells vs. controls, independently by the treatment applied. In particular, the antibacterial program was associated with lower levels of CFUs. The quantification of the metabolic activity was significantly lower by using the oxidative stress program. Live/dead images showed that C.M.F.s significantly decreased the viability of C. albicans. C.M.F.s inhibited C. albicans virulence traits reducing hyphal morphogenesis, adhesion, and biofilm formation on titanium discs. The MTS assay showed no negative effects on the viability of HGF. Independent of the adopted protocol, C.M.F.s exert antifungal and anti-virulence action against C. albicans, no cytotoxicity effects on HGF and can be useful in the prevention and treatment of yeast biofilm infections.

Chronic wound infections represent an important health problem due to the reduced response to antimicrobial treatment of the pathogens organized in structured biofilms. This study investigated the effects of the previously described antifungal peptide L18R against three representative wound pathogens: Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. The antimicrobial activity of L18R was evaluated (i) against single planktonic microbial populations; (ii) on single, dual, and triadic species of biofilms in both the early stage and mature stage; and (iii) in the polymicrobial Lubbock chronic wound biofilm (LCWB) model, mimicking spatial microbial colonization. This study used the evaluation of CFUs, biofilm biomass detection, and confocal and scanning electron microscopy analysis. L18R showed a significant antimicrobial activity against planktonic microorganisms and was able to differentially reduce the biomass of monomicrobial biofilms. No reduction of biomass was observed against the polymicrobial biofilm. In mature LCWB, L18R caused a moderate reduction in total CFU number, with a variable effect on the different microorganisms. Microscopy images confirmed a predominant presence of P.aeruginosa and a lower percentage of C. albicans cells. These findings suggest a modulating action of L18R and recommend further studies on its potential role in chronic wound management in association with conventional antibiotics or alternative treatments.

(1) Background: The objectives of this study were to evaluate the effect of several sessions of the antibacterial protocol of complex electromagnetic fields (CMFs) on planktonic Candida albicans and fungal ability, after treatment with CMFs, to adhere and proliferate on acrylic resin materials. (2) Methods: Planktonic overnight cultures of Candida albicans were subjected to different entities of CMFs treatments. Four test groups were compared: “p1”: treated only with the first program of the antibacterial protocol; “p1–p5” subjected to the first five programs; “1 antibacterial” received one complete session of the protocol and “2 antibacterial” received two complete sessions. After the treatments, the number of colony forming units (CFUs) were recorded. Then, C. albicans broth cultures were cultivated on polyacrylic resin discs and evaluated for CFUs and subjected to scanning electron microscope (SEM) analysis. (3) Results: Microbiological analysis showed that CMFs promoted a significant reduction of C. albicans CFUs when the protocol “p1–p5” was applied. No statistically significant differences between test groups were observed if the time of exposure to CMFs was increased. SEM observations and CFUs showed that CMFs treatments have the ability to reduce C. albicans adherence and proliferation on discs. (4) Conclusions: The CMFs showed an antifungal effect as well as a decrease in C. albicans adhesion on polyacrylic resin.

Athletes’ oral health appears to be poor in numerous sport activities and different diseases can limit athletic skills, both during training and during competitions. Sport activities can be considered a risk factor, among athletes from different sports, for the onset of oral diseases, such as caries with an incidence between 15% and 70%, dental trauma 14–70%, dental erosion 36%, pericoronitis 5–39% and periodontal disease up to 15%. The numerous diseases are related to the variations that involve the ecological factors of the oral cavity such as salivary pH, flow rate, buffering capability, total bacterial count, cariogenic bacterial load and values of secretory Immunoglobulin A. The decrease in the production of S-IgA and the association with an important intraoral growth of pathogenic bacteria leads us to consider the training an “open window” for exposure to oral cavity diseases. Sports dentistry focuses attention on the prevention and treatment of oral pathologies and injuries. Oral health promotion strategies are needed in the sports environment. To prevent the onset of oral diseases, the sports dentist can recommend the use of a custom-made mouthguard, an oral device with a triple function that improves the health and performance of athletes. During training, the sports dentist must monitor the athletes and the sports examination protocol must be implemented with the inclusion of the clinical examination, quantitative and qualitative analysis of saliva and instructions on the use, cleansing and storage of the mouthguard.

The drug-resistance phenomenon in Helicobacter pylori underlines the need of novel strategies to improve the eradication rate including alternative treatments combining antibiotic and non-antibiotic compounds with synergistic action. In this study, the antibacterial (MIC/MBC) and anti-virulence effects (biofilm reduction and swarming motility inhibition) of resveratrol-RSV and new synthetized RSV-phenol derivatives, with a higher bioavailability, alone and combined with levofloxacin-LVX were evaluated against resistant H. pylori clinical strains. The experiments were confirmed in vivo using the Galleria mellonella model. Among the studied RSV derivatives, RSV-3 and RSV-4 possessed higher antibacterial activity with respect to RSV (MICs from 6.25 to 200 µg/mL and from 3.12 to 200 µg/mL, respectively). RSV, RSV-3, and RSV-4 were able to synergize with LVX restoring its effect in two out of seven clinical resistant strains tested for the study. RSV, RSV-3, and RSV-4, alone and with LVX at sub-MIC and sub-synergistic concentrations, significantly reduced the biofilm formation. Moreover, RSV-3 and RSV-4 reduced the H. pylori swarming motility on soft agar. RSV, RSV-3, and RSV-4 were non-toxic for G. mellonella larvae and displayed a protective effect against H. pylori infection. Overall, RSV–phenol derivatives should be considered interesting candidates for innovative therapeutic schemes to tackle the H. pylori antibiotic resistance.

Background: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria can be efficiently tackled by using prebiotics. The aim of this work was to identify novel prebiotic combinations by exerting species-specific action between S. aureus and S. epidermidis strains. Methods: First, the antimicrobial/antibiofilm effect of Xylitol-XYL and Galacto-OligoSaccharides–GOS combined with each other at different concentrations (1, 2.5, 5%) against S. aureus and S. epidermidis clinical strains was evaluated in time. Second, the most species-specific concentration was used to combine XYL with Fructo-OligoSaccharides–FOS, IsoMalto-Oligosaccharides–IMO, ArabinoGaLactan–LAG, inulin, dextran. Experiments were performed by OD600 detection, biomass quantification and LIVE/DEAD staining. Results: 1% XYL + 1% GOS showed the best species-specific action with an immediate antibacterial/antibiofilm action against S. aureus strains (up to 34.54% ± 5.35/64.68% ± 4.77) without a relevant effect on S. epidermidis. Among the other prebiotic formulations, 1% XYL plus 1% FOS (up to 49.17% ± 21.46/37.59% ± 6.34) or 1% IMO (up to 41.28% ± 4.88/36.70% ± 10.03) or 1% LAG (up to 38.21% ± 5.31/83.06% ± 5.11) showed antimicrobial/antibiofilm effects similar to 1% XYL+1% GOS. For all tested formulations, a prevalent bacteriostatic effect in the planktonic phase and a general reduction of S. aureus biofilm formation without loss of viability were recorded. Conclusion: The combinations of 1% XYL with 1% GOS or 1% FOS or 1% IMO or 1% LAG may help to control the balance of skin microbiota, representing good candidates for topic formulations.

This work aimed to compare the capability of Streptococcus oralis to adhere to a novel surface, double-etched titanium (DAE), in respect to machined and single-etched titanium. The secondary outcome was to establish which topographical features could affect the interaction between the implant surface and bacteria. The samples’ superficial features were characterized using scanning electron microscopy (SEM) and energy dispersive x-ray spectrometry (EDS), and the wetting properties were tested through sessile methods. The novel surface, the double-etched titanium (DAE), was also analyzed with atomic force microscopy (AFM). S. oralis was inoculated on discs previously incubated in saliva, and then the colony-forming units (CFUs), biomass, and cellular viability were measured at 24 and 48h. SEM observation showed that DAE was characterized by higher porosity and Oxygen (%) in the superficial layer and the measurement of the wetting properties showed higher hydrophilicity. AFM confirmed the presence of a higher superficial nano-roughness. Microbiological analysis showed that DAE discs, coated by pellicle’s proteins, were characterized by significantly lower CFUs at 24 and 48 h with respect to the other two groups. In particular, a significant inverse relationship was shown between the CFUs at 48 h and the values of the wetted area and a direct correlation with the water contact angle. The biomass at 24 h was slightly lower on DAE, but results were not significant concerning the other groups, both at 24 and 48 h. The DAE treatment not only modifies the superficial topography and increased hydrophilicity, but it also increases the Oxygen percentage in the superficial layer, which could contribute to the inhibition of S. oralis adhesion. DAE can be considered a promising treatment for titanium implants to counteract a colonization pioneer microorganism, such as S. oralis.

ABSTRACT Young swimmers are particularly susceptible to the onset of oral diseases. Objective To evaluate the oral health status in young competitive and non-competitive swimmers, involving an assessment of salivary cariogenic bacteria and secretory IgA (S-IgA) concentration. Material and Methods Before training sessions (T1), 54 competitive and 69 non-competitive swimmers had the following parameters assessed: decayed, missing, and filled teeth (DMFT), Plaque Index (PlI), and Gingival Index (GI). At T1 and after training sessions (T2), stimulated saliva was collected and microbiological and immunological analyses were performed. Results Competitive swimmers trained 2.02±0.09 hours 5 times a week, while non-competitive swimmers trained 2.03±0.18 hours a week. A total of 14.7% of competitive swimmers suffered dental trauma related to sports. Only 11.76% of the competitive swimmers took a daily dose of fluoride, against 32.65% of non-competitive swimmers (p=0.029). Neither group followed an established diet or presented statistically significant differences in terms of nutritional supplement drink and chocolate intake. There were statistically significant differences in terms of oral hygiene. No significant difference in clinical indexes (DMFT, PlI, and GI) was present. S. mutans was harbored by 18.6% of competitive and the 32.2% of non-competitive swimmers. S. sobrinus was detected in 22.03% of competitive and 91.6% of non-competitive swimmers (p

Background/purpose: A bacterial adhesion to implant surfaces is a first stage of peri-implant mucositis and peri-implantitis. The aim of this longitudinal study was to examine the quantitative and qualitative biofilm formation on healing screws and the presence of periodontal pathogens in peri-implant crevicular fluid (PCF). In each of the 30 participants, one healing screw was adapted to every single implant. Materials and methods: Thirty patients, 18 females and 12 males, were selected. Thirty healing abutments were left in situ for 20, 30 and 90 days. At regular times, the presence/absence of bleeding on probing (BOP) was determined. The specific periodontal pathogens were determined in PCF, by polymerase chain reaction (PCR). After 20, 30 and 90 days, the healing screws were removed and analyzed to establish the total bacterial count by a culture method. Results: BOP+ increased significantly after 90 days. Biofilm was detected on all healing abutments and the number of cultivable oral flora showed a significant increase from 20 days to 30 days and to 90 days. At 90 days, the anaerobic counts constituted the bulk of plaque examined and Veillonella spp. were present in higher percentages levels (19.82%) among the Gram-negative bacteria. No significant differences among the experimental groups were detected in the frequencies of detection of each monitored bacteria in PCF. Conclusion: The healing screws left in situ for a period of 90 days caused a peri-implant inflammation and the presence of periodontal pathogenic bacteria in the peri-implant sulcus, due to the plaque accumulation on screw surfaces.

Aim Bacterial adherence to implants is considered to be an important event in the pathogenesis of bacterial infections. In fact, this infection process is a first stage of peri-implant mucositis and peri-implantitis, and a positive correlation has been found between oral hygiene and marginal bone loss around implants in the edentulous mandible. Surface properties of transgingival implant components are important determinants in bacterial adhesion. The purpose of this study was to characterize the biofilm formation, in vivo, on healing screws made of titanium alloy or coated with a combination of anatase and Bactercline® product. Materials and methods Twenty-five patients, between 21- 37 years, in excellent systemic health, participated in this study. In each of the 25 participants, one anatase-Bactercline® coated healing screw (Test) and one titanium alloy (TI6Al4V) healing screw (Control) were adapted to two different implants. Quantitative and qualitative biofilm formation on healing abutments was analyzed by culture method.Results Bacterial adherence to the two different healing screws used in this study were compared. Statistically significant differences were found between the Control and the Test group for both aerobic and anaerobic bacterial counts (p