The race for the optimal antimicrobial surface: perspectives and challenges related to plasma electrolytic oxidation coating for titanium-based implants.

Plasma electrolytic oxidation (PEO) is a low-cost, structurally reliable, and environmentally friendly surface modification method for orthopedic and dental implants. This technique is successful for the formation of porous, corrosion-resistant, and bioactive coatings, besides introducing antimicrobial compounds easily. Given the increase in implant-related infections, antimicrobial PEO-treated surfaces have been widely proposed to surmount this public health concern. This review comprehensively discusses antimicrobial implant surfaces currently produced by PEO in terms of their in vitro and in vivo microbiological and biological properties. We present a critical [part I] and evidence-based [part II] review about the plethora of antimicrobial PEO-treated surfaces. The mechanism of microbial accumulation on implanted devices and the principles of PEO technology to ensure antimicrobial functionalization by one- or multi-step processes are outlined. Our systematic literature search showed that particular focus has been placed on the metallic and semi-metallic elements incorporated into PEO surfaces to facilitate antimicrobial properties, which are often dose-dependent, without leading to cytotoxicity in vitro. Meanwhile, there are concerns over the biocompatibility of PEO and its long-term antimicrobial effects in animal models. We clearly highlight the importance of using clinically relevant infection models and in vivo long-term assessments to guarantee the rational design of antimicrobial PEO-treated surfaces to identify the 'finish line' in the race for antimicrobial implant surfaces.

Tailoring Cu2+-loaded electrospun membranes with antibacterial ability for guided bone regeneration.

Copper (Cu)-loaded electrospun membranes were tailored for guided bone regeneration (GBR), targeting the stimulation of innate cells active in bone growth and the prevention of bacterial infections. Functional GBR membranes were produced via an electrospinning set-up using a silk-based solution associated with polyethylene oxide (Silk/PEO - control). Experimental groups were loaded with copper oxide using varying weight percentages (0.05 % to 1 % of CuO). The morphological, structural, chemical, and mechanical properties of membranes were evaluated. Direct and indirect in vitro cytocompatibility experiments were performed with primary human bone mesenchymal stem cells and primary human umbilical vein endothelial cells. The antibacterial potential of membranes was tested with Staphylococcus aureus and Fusobacterium nucleatum biofilm. CuO was successfully incorporated into membranes as clusters without compromising their mechanical properties for clinical applicability. Increased Cu concentrations generated membranes with thinner nanofibers, greater pore areas, and stronger antimicrobial effect (p < 0.01). Cu2+ ion was released from the nanofiber membranes during 1 week, showing higher release in acidic conditions. CuO 0.1 % and CuO 0.05 % membranes were able to support and stimulate cell adhesion and proliferation (p < 0.05), and favor angiogenic responses of vascular cells. In addition, detailed quantitative and qualitative analysis determined that amount of the attached biofilm was reduced on the tailored functional Cu2+-loaded GBR membrane. Importantly, these qualities represent a valuable strategy to improve the bone regeneration process and diminish the risk of bacterial infections.

Copper source determines chemistry and topography of implant coatings to optimally couple cellular responses and antibacterial activity.

Implant-related infections at the early healing period are considered one of the main risk factors in implant failure. Designing coatings that control bacterial adhesion and have cell stimulatory behavior remains a challenging strategy for dental implants. Here, we used plasma electrolytic oxidation (PEO) to produce antimicrobial coatings on commercially pure titanium (cpTi) using bioactive elements (calcium and phosphorus) and different copper (Cu) sources: copper acetate (CuAc), copper sulfate (CuS), and copper oxide (CuO); coatings containing only Ca and P (CaP) served as controls. Cu sources drove differential physical and chemical surface features of PEO coatings, resulting in tailorable release kinetics with a sustained Cu ion release over 10 weeks. The antibacterial effects of Cu-containing coatings were roughness-dependent. CuAc coating exhibited optimal properties in terms of its hydrophilicity, pores density, and limited surface roughness, which provided the most robust antibacterial activity combined with appropriate responses of human primary stem cells and angiogenic cells. Our data indicate that Cu source selection largely determines the functionality of Cu-containing PEO coatings regarding their antibacterial efficacy and cytocompatibility.

Surface Engineering for Dental Implantology: Favoring Tissue Responses Along the Implant.

Dental implants represent an illustrative example of successful medical devices used in increasing numbers to aid (partly) edentulous patients. Particularly in spite of the percutaneous nature of dental implant systems, their clinical success is remarkable. This clinical success is at least partly related to the effective surface treatment of the artificial dental root, providing appropriate physicochemical properties to achieve osseointegration. The demographic changes in the world, however, with a rapidly increasing life expectancy and an increase in patients suffering from comorbidities that affect wound healing and bone metabolism, make that the performance of dental implants requires continuous improvement. An additional factor endangering the clinical success of dental implants is peri-implantitis, which affects both the soft and hard tissue interactions with dental implants. In this study, we shed light on the optimization of dental implant surfaces through surface engineering. Depending on the region along the artificial dental root, different properties of the surface are required to optimize prevailing tissue response to facilitate osseointegration, improve soft tissue attachment, and exert antibacterial efficacy. As such, surface engineering represents an important tool for assuring the continued future success of dental implants. Impact Statement Dental implants represent a common treatment modality nowadays for the replacement of lost teeth or fixation of prosthetic devices. This review provides a detailed overview of the role of surface engineering for dental implants and their components to optimize tissue responses at the different regions along the artificial dental root. The surface properties steering immunomodulatory processes, facilitating osseointegration, and rendering antibacterial efficacy (at both artificial root and abutment region) are described. The review finally concludes that surface engineering provides a tool to warrant that dental implants will remain future proof in more challenging applications, including an aging patient population and comorbidities that affect bone metabolism and wound healing.

Insight Into Corrosion of Dental Implants: From Biochemical Mechanisms to Designing Corrosion-Resistant Materials.

Purpose of Review: Despite advanced technologies to avoid corrosion of dental implants, the mechanisms toward the release of metals and their role in the onset of peri-implant diseases are still under-investigated. Effective knowledge on the etiopathogenesis of corrosive products and preventive strategies mitigating the risks for surface degradation are thus in dire need. This review aimed to summarize evidence toward biocorrosion in the oral environment and discuss the current strategies targeting the improvement of dental implants and focusing on the methodological and electrochemical aspects of surface treatments and titanium-based alloys. Recent Findings: Recent studies suggest the existence of wear/corrosion products may correlate with peri-implantitis progress by triggering microbial dysbiosis, the release of pro-inflammatory cytokines, and animal bone resorption. Furthermore, current clinical evidence demonstrating the presence of metal-like particles in diseased tissues supports their possible role as a risk factor for peri-implantitis. For instance, to overcome the drawback of titanium corrosion, researchers are primarily focusing on developing corrosion-resistant alloys and coatings for dental implants by changing their physicochemical features. Summary: The current state-of-art discussed in this review found corrosion products effective in affecting biofilm virulence and inflammatory factors in vitro. Controversial and unstandardized data are limitations, making the premise of corrosion products being essential for peri-implantitis onset. On the other hand, when it comes to the strategies toward reducing implant corrosion rate, it is evident that the chemical and physical properties are crucial for the in vitro electrochemical behavior of the implant material. For instance, it is foreseeable that the formation of films/coatings and the incorporation of some functional compounds into the substrate may enhance the material's corrosion resistance and biological response. Nevertheless, the utmost challenge of research in this field is to achieve adequate stimulation of the biological tissues without weakening its protective behavior against corrosion. In addition, the translatability from in vitro findings to clinical studies is still in its infancy. Therefore, further accumulation of high-level evidence on the role of corrosion products on peri-implant tissues is expected to confirm the findings of the present review besides the development of better methods to improve the corrosion resistance of dental implants. Furthermore, such knowledge could further develop safe and long-term implant rehabilitation therapy.

Optimizing citric acid protocol to control implant-related infections: An in vitro and in situ study.

OBJECTIVE: The present study aimed to establish an optimized protocol for biofilm removal from titanium (Ti) surfaces using citric acid (CA) solutions. BACKGROUND: Biofilm accumulation is the main factor to trigger peri-implant infections and to increase the risk of treatment failures. Although CA has been suggested as the anti-infective agent with highest potential for biofilm removal on Ti, there is no consensus that CA could improve the anti-infective treatment and its effect. METHODS: Physical and chemical alterations, electrochemical behavior, cytotoxicity, and antimicrobial effect of CA on Ti discs were evaluated using four concentrations (1, 10, 20, and 40%) and two application methods (immersion and rubbing). Negative control using 0.9% NaCl was used in all experiments. To evaluate whether different application times can have similar response, polymicrobial biofilm (microcosm model) was formed on Ti and treated with CA for 1, 2, 4, and 8 min. An in situ study was conducted to verify whether the established protocol is equally effective in biofilms formed on machined and sandblasted, large-grit, and acid-etched (SLA) Ti surfaces. RESULTS: CA 40% induced significantly higher surface alterations observed by confocal images and profilometry. In general, rubbing protocol decreased the surface roughness and increased the wettability (p < 0.05), exhibiting better surface cleaning by biofilm removal. CA 10% presented no indirect cytotoxicity and, when applied by rubbing for 8 min, presented proper in vitro antibacterial action and potential corrosion inhibition. When CA 10% was rubbed on Ti surfaces for 4 min, it displayed optimum cleaning ability as 8 min, working equally to remove in situ biofilm on machined and SLA surfaces. CONCLUSIONS: The application of CA 10% by rubbing for at least 4 min demonstrated to be a promising protocol to eliminate biofilms formed in smooth and rougher surfaces, which could improve implant-related infection therapies.

Synthesis of bioactive glass-based coating by plasma electrolytic oxidation: Untangling a new deposition pathway toward titanium implant surfaces.

HYPOTHESIS: Although bioactive glass (BG) particle coatings were previously developed by different methods, poor particle adhesion to surfaces and reduced biological effects because of glass crystallization have limited their biomedical applications. To overcome this problem, we have untangled, for the first time, plasma electrolytic oxidation (PEO) as a new pathway for the synthesis of bioactive glass-based coating (PEO-BG) on titanium (Ti) materials. EXPERIMENTS: Electrolyte solution with bioactive elements (Na2SiO3-5H2O, C4H6O4Ca, NaNO3, and C3H7Na2O6P) was used as a precursor source to obtain a 45S5 bioglass-like composition on a Ti surface by PEO. Subsequently, the PEO-BG coating was investigated with respect to its surface, mechanical, tribological, electrochemical, microbiological, and biological properties, compared with those of machined and sandblasted/acid-etched control surfaces. FINDINGS: PEO treatment produced a coating with complex surface topography, Ti crystalline phases, superhydrophilic status, chemical composition, and oxide layer similar to that of 45S5-BG (~45.0Si, 24.5 Ca, 24.5Na, 6.0P w/v%). PEO-BG enhanced Ti mechanical and tribological properties with higher corrosion resistance. Furthermore, PEO-BG had a positive influence in polymicrobial biofilms, by reducing pathogenic bacterial associated with biofilm-related infections. PEO-BG also showed higher adsorption of blood plasma proteins without cytotoxic effects on human cells, and thus may be considered a promising biocompatible approach for biomedical implants.

UV-photofunctionalization of a biomimetic coating for dental implants application.

Photofunctionalization mediated by ultraviolet (UV) rays changes the physico-chemical characteristics of titanium (Ti) and improves the biological activity of dental implants. However, the role of UV-mediated photofunctionalization of biofunctional Ti surfaces on the antimicrobial and photocatalytic activity remains unknown and was investigated in this study. Commercially pure titanium (cpTi) discs were divided into four groups: (1) machined samples without UV light application [cpTi UV-]; (2) plasma electrolytic oxidation (PEO) treated samples without UV light application [PEO UV-]; (3) machined samples with UV light application [cpTi UV+]; and (4) PEO-treated samples with UV light application [PEO UV+]. The surfaces were characterized according to their morphology, roughness, crystalline phase, chemical composition and wettability. The photocatalytic activity and proteins adsorption were measured. For the microbiological assay, Streptococcus sanguinis was grown on the disc surfaces for 1 h and 6 h, and the colony forming units and bacterial organization were evaluated. In addition, to confirm the non-cytotoxic effect of PEO UV +, human gingival fibroblast (HGF) cells were cultured in a monolayer onto each material surface and the cells viability and proliferation evaluated by a fluorescent cell staining method. PEO treatment increased the Ti surface roughness and wettability (p < 0.05). Photofunctionalization reduced the hydrocarbon concentration and enhanced human blood plasma proteins and albumin adsorption mainly for the PEO-treated surface (p < 0.05). PEO UV+ also maintained higher wettability values for a longer period and provided microbial reduction at 1 h of bacterial adhesion (p = 0.012 vs. PEO UV-). Photofunctionalization did not increase the photocatalytic activity of Ti (p > 0.05). Confocal microscopy analyses demonstrated that PEO UV+ had no cell damage effect on HGF cells growth even after 24 h of incubation. The photofunctionalization of a biofunctional PEO coating seems to be a promising alternative for dental implants as it increases blood plasma proteins adsorption, reduces initial bacterial adhesion and presents no cytotoxicity effect.

Targeting Pathogenic Biofilms: Newly Developed Superhydrophobic Coating Favors a Host-Compatible Microbial Profile on the Titanium Surface.

Polymicrobial infections are one of the most common reasons for inflammation of surrounding tissues and failure of implanted biomaterials. Because microorganism adhesion is the first step for biofilm formation, physical-chemical modifications of biomaterials have been proposed to reduce the initial microbial attachment. Thus, the use of superhydrophobic coatings has emerged because of their anti-biofilm properties. However, these coatings on the titanium (Ti) surface have been developed mainly by dual-step surface modification techniques and have not been tested using polymicrobial biofilms. Therefore, we developed a one-step superhydrophobic coating on the Ti surface by using a low-pressure plasma technology to create a biocompatible coating that reduces polymicrobial biofilm adhesion and formation. The superhydrophobic coating on Ti was created by the glow discharge plasma using Ar, O2, and hexamethyldisiloxane gases, and after full physical, chemical, and biological characterizations, we evaluated its properties regarding oral biofilm inhibition. The newly developed coating presented an increased surface roughness and, consequently, superhydrophobicity (contact angle over 150°) and enhanced corrosion resistance (p < 0.05) of the Ti surface. Furthermore, proteomic analysis showed a unique pattern of protein adsorption on the superhydrophobic coating without drastically changing the biologic processes mediated by proteins. Additionally, superhydrophobic treatment did not present a cytotoxic effect on fibroblasts or reduction of proliferation; however, it significantly reduced (≈8-fold change) polymicrobial adhesion (bacterial and fungal) and biofilm formation in vitro. Interestingly, superhydrophobic coating shifted the microbiological profile of biofilms formed in situ in the oral cavity, reducing by up to ≈7 fold pathogens associated with the peri-implant disease. Thus, this new superhydrophobic coating developed by a one-step glow discharge plasma technique is a promising biocompatible strategy to drastically reduce microbial adhesion and biofilm formation on Ti-based biomedical implants.

Visible-Light-Induced Photocatalytic and Antibacterial Activity of TiO2 Codoped with Nitrogen and Bismuth: New Perspectives to Control Implant-Biofilm-Related Diseases.

Biofilm-associated diseases are one of the main causes of implant failure. Currently, the development of implant surface treatment goes beyond the osseointegration process and focuses on the creation of surfaces with antimicrobial action and with the possibility to be re-activated (i.e., light source activation). Titanium dioxide (TiO2), an excellent photocatalyst used for photocatalytic antibacterial applications, could be a great alternative, but its efficiency is limited to the ultraviolet (UV) range of the electromagnetic spectrum. Since UV radiation has carcinogenic potential, we created a functional TiO2 coating codoped with nitrogen and bismuth via the plasma electrolytic oxidation (PEO) of titanium to achieve an antibacterial effect under visible light with re-activation potential. A complex surface topography was demonstrated by scanning electron microscopy and three-dimensional confocal laser scanning microscopy. Additionally, PEO-treated surfaces showed greater hydrophilicity and albumin adsorption compared to control, untreated titanium. Bismuth incorporation shifted the band gap of TiO2 to the visible region and facilitated higher degradation of methyl orange (MO) in the dark, with a greater reduction in the concentration of MO after visible-light irradiation even after 72 h of aging. These results were consistent with the in vitro antibacterial effect, where samples with nitrogen and bismuth in their composition showed the greatest bacterial reduction after 24 h of dual-species biofilm formation ( Streptococcus sanguinis and Actinomyces naeslundii) in darkness with a superior effect at 30 min of visible-light irradiation. In addition, such a coating presents reusable photocatalytic potential and good biocompatibility by presenting a noncytotoxicity effect on human gingival fibroblast cells. Therefore, nitrogen and bismuth incorporation into TiO2 via PEO can be considered a promising alternative for dental implant application with antibacterial properties in darkness, with a stronger effect after visible-light application.

Citric acid reduces oral biofilm and influences the electrochemical behavior of titanium: An in situ and in vitro study.

BACKGROUND: Citric acid (CA) has been suggested as an effective antimicrobial agent against biofilms formed on titanium. However, the antimicrobial effect of CA in biofilms formed in the oral environment and its effects on the physical properties of titanium surface remain unknown. Therefore, this study evaluated the antimicrobial effect of CA on in situ biofilm, whether this treatment favors the bacteria recolonization and its effect on the electrochemical properties of titanium. METHODS: In the in situ test, volunteers wore a palatal appliance containing titanium discs. After 7 days, the discs (N = 21) were exposed in vitro to the following treatments: immersion in 0.9% sodium chloride (control); 40% CA immersion; and 40% CA rubbing. Afterwards, these discs (N = 18) were exposed in vitro to new bacterial adhesion with Streptococcus sanguinis. New discs (N = 18) without biofilm were exposed to the treatments and subjected to electrochemical tests and surface characterization. Data were analyzed by one-way ANOVA followed by Tukey's HSD test. RESULTS: The CA groups showed a significant reduction (≈ 5-log reduction) in the biofilm formed in situ compared with the control group (p < 0.05), but no difference was found between CA application methods (p = 0.680). The acid treatment did not favor the recolonization of bacteria (p = 0.629). CA treatment did not influence the polarization resistance and capacitance of the oxide film, but statistically enhanced the electrochemical stability of titanium. CONCLUSION: Citric acid appears to be an effective clinical alternative for treatment of the main etiologic factor in dental implant failure, biofilm formation, enhancing electrochemical behavior of titanium.

Characterization of chemically treated Ti-Zr system alloys for dental implant application.

Materials and surfaces developed for dental implants need to withstand degradation processes that take place in the oral cavity. Therefore, the aim of the study was to develop and evaluate the topographical, mechanical, chemical, electrochemical and biological properties of Ti-xZr alloys (x = 5, 10, and 15 wt%) with two surface features (machined and double acid etched). Commercially pure titanium (cpTi) and Ti-6Al-4V alloy were used as controls. Surface characterization was performed using dispersive energy spectroscopy, X-ray diffraction, scanning electron microscopy, atomic force microscopy, profilometry and surface energy. The mechanical properties were assessed using Vickers microhardness, elastic modulus and stiffness. The electrochemical behavior analysis was conducted in a body fluid solution (pH 7.4). In addition, MC3T3-E1 cells were used to determine the impact of material and surface treatment on cell morphology by SEM analysis. Data were analyzed by two-way ANOVA and Bonferroni test (α = 0.05). Ti-Zr alloys showed lower surface roughness, elastic modulus and stiffness, as well as higher hardness and surface energy when compared to cpTi. Ti-Zr system increased the polarization resistance values and significantly decreased the capacitance, corrosion current density (icorr), and passivation current density (ipass) values. The acid treatment increased the resistance and corrosion potential of the oxide layer. SEM data analysis demonstrated that Ti-Zr alloys displayed normal cell attachment/spreading and slightly changed cell morphology in the double etched surface. In conclusion, Zr addition and surface treatment altered surface, mechanical, biological and electrochemical properties of Ti material.

Development of binary and ternary titanium alloys for dental implants.

OBJECTIVE: The aim of this study was to develop binary and ternary titanium (Ti) alloys containing zirconium (Zr) and niobium (Nb) and to characterize them in terms of microstructural, mechanical, chemical, electrochemical, and biological properties. METHODS: The experimental alloys - (in wt%) Ti-5Zr, Ti-10Zr, Ti-35Nb-5Zr, and Ti-35Nb-10Zr - were fabricated from pure metals. Commercially pure titanium (cpTi) and Ti-6Al-4V were used as controls. Microstructural analysis was performed by means of X-ray diffraction and scanning electron microscopy. Vickers microhardness, elastic modulus, dispersive energy spectroscopy, X-ray excited photoelectron spectroscopy, atomic force microscopy, surface roughness, and surface free energy were evaluated. The electrochemical behavior analysis was conducted in a body fluid solution (pH 7.4). The albumin adsorption was measured by the bicinchoninic acid method. Data were evaluated through one-way ANOVA and the Tukey test (α=0.05). RESULTS: The alloying elements proved to modify the alloy microstructure and to enhance the mechanical properties, improving the hardness and decreasing the elastic modulus of the binary and ternary alloys, respectively. Ti-Zr alloys displayed greater electrochemical stability relative to that of controls, presenting higher polarization resistance and lower capacitance. The experimental alloys were not detrimental to albumin adsorption. SIGNIFICANCE: The experimental alloys are suitable options for dental implant manufacturing, particularly the binary system, which showed a better combination of mechanical and electrochemical properties without the presence of toxic elements.

Is there scientific evidence favoring the substitution of commercially pure titanium with titanium alloys for the manufacture of dental implants?

The development of Ti alloys to manufacture dental implants has emerged in recent years due to the increased failure of commercially pure titanium (cpTi) implants. Thus, this study reviews existing information about the mechanical, chemical, electrochemical, and biological properties of the main Ti alloys developed over the past few years to provide scientific evidence in favor of using Ti-based alloys as alternative to cpTi. Ti alloys may be considered viable substitutes in the fabrication of dental implants. Such evidence is given by the enhanced properties of alloys, such as a low elastic modulus, high tensile strength, satisfactory biocompatibility, and good corrosion and wear resistances. In addition, Ti alloys may be modified at the structural, chemical, and thermomechanical levels, which allows the development of materials in accordance with the demands of several situations encountered in clinical practice. Although several in vitro studies have established the superiority of Ti alloys over cpTi, mainly in terms of their mechanical properties, there is no scientific evidence that supports the total replacement of this material in vivo. This review demonstrates the superiority of ß-type alloys. However, it is evident that in vivo studies are encouraged to test new alloys to consolidate their use as substitutes for cpTi.