Background: antimicrobial resistance represents a critical issue leading to delayed wound healing; hence, it is necessary to develop novel strategies to address this phenomenon. Objectives: this study aimed to explore the antimicrobial/anti-virulence action of Methylglyoxal-MGO alone or combined with novel technologies such as Light-Emitting Diodes-LED and Complex Magnetic Fields-CMFs against resistant clinical strains isolated from chronic wounds. Methods: characterized planktonic Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans isolates were used. Antimicrobial activity was evaluated by measuring optical density, Colony Forming Units-CFU, and synergy between MGO/LED or CMFs. Cellular membrane permeability by propidium iodide fluorescence and fluidity by Laurdan generalized polarization measurements were performed. P. aeruginosa motility was tested using the soft agar method. A docking study was performed to evaluate the possible interaction between MGO and urease in P. aeruginosa. Results: single/combined treatments showed significant antimicrobial activity. Major CFU reduction was detected after CMFs/MGO+CMFs application on C. albicans. Treatments exhibited significant changes in membrane permeability and fluidity. The treatments decreased P. aeruginosa motility with a major reduction after LED application. Docking analysis showed that MGO could bind with P. aeruginosa urease leading to defective folding and functional alterations. Conclusions: the results suggest that these treatments could represent promising and green therapeutic solutions against resistant isolates from chronic wounds.

The increase in multi-drug-resistant strains represents a global challenge that strongly underlines the importance of the search for new eco-sustainable strategies. The aim of this work was to suggest a non-antibiotic approach to counteract resistant Staphylococcus pseudintermedius and Pseudomonas aeruginosa strains in planktonic and sessile phases. The proposed strategy includes the combination of natural spliced variant of hepatocyte growth factor NK1, a protein produced by recombinant DNA technology in Pichia pastoris expression system and Light-Emitting Diodes (LEDs). The antimicrobial action was determined by Minimum Inhibitory Concentration, Minimum Bactericidal Concentration, and CFU/ml evaluations. The anti-virulence action was performed by measuring P. aeruginosa motility and twitching and anti-S. pseudintermedius and -P. aeruginosa biofilms. Recombinant NK1 and LEDs alone and combined with each other showed relevant antimicrobial and anti-virulence effects. In particular, when NK1 was combined with LEDs, significant CFU/ml reduction (up to 85.98% reduction with respect to the control) after 24 h of contact, P. aeruginosa swimming/swarming/twitching halos reduction (up to 60% reduction with respect to the control) and bacterial anti-biofilm formation action was obtained. The obtained results demonstrated the innovative and interesting non-antibiotic strategy formed by recombinant NK1 protein and LEDs to affect the S. pseudintermedius and P. aeruginosa growth.

Chronic wound infections associated with resistant polymicrobial biofilms are often refractory to conventional therapies with sustained healing time. This study evaluated the efficacy of non-antibiotic treatments including Methylglyoxal-MGO-Light-Emitting Diode-LED-and Complex Magnetic Fields-CMFs-alone/combined against the biofilms of two polymicrobial mixes (MIX 1, MIX 2) containing S. aureus, P. aeruginosa and C. albicans using the Lubbock chronic wound biofilm model. At 24 h after treatment, the effects were evaluated by (i) CFU/mg reduction, (ii) Confocal Laser Scanning Microscopy-CLSM-and (iii) Scanning Electron Microscopy-SEM. All treatments significantly reduced biofilms in terms of CFU/mg in both mixes versus the controls, 24 h after treatment. MGO showed remarkable activity, especially against P. aeruginosa. In MIX 1, LED/MGO + LED was highly effective against C. albicans. The combinations MGO + LED/MGO + CMFs enhanced the antibiofilm activity compared to each single treatment against S. aureus and P. aeruginosa, in both MIX1/MIX2. CLSM and SEM analysis showed biofilm disaggregation and reduction in cell viability with combined treatments, and Candida hyphal inhibition after CMFs application. In conclusion, the results demonstrate that MGO, alone or combined with LED or CMFs, shows high efficacy against resistant biofilms in the LCWB model 24 h after treatment, and encourage further studies on potential non-antibiotic and eco-friendly strategies as future alternative therapeutic approaches for chronic wound infections.

AIM:Bacterial biofilms can form on surfaces in hospitals, clinics, farms, and food processing plants, representing a possible source of infections and cross-contamination. This study investigates the effectiveness of new commercial wipes against Staphylococcus aureus and Pseudomonas aeruginosa biofilms (early attachment and formed biofilms), assessing LH SALVIETTE wipes (Lombarda H S.r.l.) potential for controlling biofilm formation. METHODS AND RESULTS:The wipes efficacy was studied against the early attachment phase and formed biofilm of S. aureus ATCC 6538 and P. aeruginosa ATCC 15442 on a polyvinyl chloride (PVC) surface, following a modified standard test EN 16615:2015, measuring Log10 reduction and cell viability using live/dead staining. It was also evaluated the wipes anti-adhesive activity over time (3 h, 2 4h), calculating CFU.mL-1 reduction. Data were analyzed using t-student test. The wipes significantly reduced both early phase and formed S. aureus biofilm, preventing dispersion on PVC surfaces. Live/dead imaging showed bacterial cluster disaggregation and killing action. The bacterial adhesive capability decreased after short-time treatment (3 h) with the wipes compared to 24 h. CONCLUSIONS:Results demonstrated decreased bacterial count on PVC surface both for early attachment phase and formed biofilms, also preventing the bacterial biofilm dispersion.

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 < 0.0001. The toluidine-blue staining observations showed the typical morphology of cells and the presence of elongated and spindle-shaped with cytoplasmic extensions and lamellipodia was observed by SEM. The ANTIBACTERIAL program statistically increased the production of collagen with respect to control and STRESS program (p < 0.0001). CMFs showed a relevant anti-virulence action against C. albicans , no cytotoxicity effects and a high hGFs migration rate. The results of this study suggest that CMFs could represent a novel eco-sustainable strategy to counteract the resistant yeast biofilm infections.

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.

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) (T 0 ) and 3 h after continuous use of the mask(s) (T 1 ). 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 T 0 and T 1 . 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 T 0 and T 1 . Conclusion It is concluded that the prolonged use of the FFP2 mask covered by a surgical mask can generate oral alterations in the user.

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.

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.

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.

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.

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.

The aim of this work was to characterize at nano-, micro-, and chemical level the surface topography of two new surfaces produced in Ti6Al4V by selective laser melting (SLM) and then subjected to the electrochemical polishing (EL) and organic acids-etching (OAE). Moreover, the capability of Streptococcus oralis to form biofilm on these surfaces was also studied. In addition, the microbiological results were compared with the topographical characteristics of the surfaces. The nano-pattering of the samples was investigated with the Atomic Force Microscopy (AFM); the micro-features, before and after the bacterial growth, were evaluated by the SEM observation and the Energy Dispersive X-ray Spectrometry (EDS) analyzed the chemical composition of the upper layers. The wetting properties of the discs were also evaluated. The colony-forming units (CFUs), the bacterial biomass, and the percentage of viable and dead cells were measured after 24 h of S. oralis biofilm formation on the different discs. Organic acids-etching samples were characterized by a ripple texture, higher nano-roughness, and wettability, respecting the machined and EL ones. The SEM observations showed that EL and OAE samples had an irregular surface with respect to the machined ones. The EDS found no significant differences for the Oxygen content of the upper layers of the samples. The microbiological analysis showed a significantly lower bacterial count and biofilm biomass formation on OAE in respect to the EL and the machined samples. In particular, the S. oralis biofilm formation was inversely correlated with the nano-roughness of the samples. The process of production of the discs, turning or SLM, significantly affected the micro-roughness of the samples. On the contrary, the treatment with OAE and EL modified the nano-topography of the samples and influencing the bacterial biofilm growth.

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.

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.

Multidrug-resistant organisms (MDRO) are bacteria that exhibit acquired resistance to multiple antibiotics, reducing the efficacy of antimicrobial therapies [...].

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 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.

PURPOSE:The aim of this study was to compare the Streptococcus oralis biofilm formation on titanium machined turned surfaces and sandblasted surfaces that were previously characterized for their superficial topographies.MATERIALS AND METHODS:Two commercially pure titanium surfaces were analyzed and compared: machined (turned surfaces subjected to a process of decontamination that also included a double acid attack) and sandblasted (sandblasted surfaces, cleaned with purified water, enzymatic detergent, acetone, and alcohol). The characterization of the samples at the nanolevel was performed using atomic force microscopy, which permitted calculation of the superficial nanoroughness (Ra). The sessile drop method was used to measure the water contact angle in both groups and allowed information to be gained about their wetting properties. Scanning electron microscope and energy-dispersive x-ray spectroscopy analysis allowed comparison of the microtopographic geometry and the chemical composition of the samples. Then, the disks were pre-incubated with saliva in order to form an acquired pellicle. Streptococcus oralis was put on the disks, and both groups were tested at 24 and 48 hours for biofilm biomass evaluation, colony-forming units (CFUs), and live/dead staining for cell viability.RESULTS:The sandblasted samples were characterized by a significantly higher level of superficial oxides, superficial roughness, and hydrophilicity, compared with the machined turned samples. Although there were topographic differences, the Streptococcus oralis biofilm formation, quantified in CFUs, and biomass formation at 24 and 48 hours were similar in both groups. With the live/dead staining, the sandblasted disks were characterized by an increased percentage of dead cells compared with the machined disks.CONCLUSION:Although significant topographic differences were present between machined and sandblasted disks, the Streptococcus oralis biofilm formation seems to not be significantly affected.

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.