Nanostructured Surfaces to Promote Osteoblast Proliferation and Minimize Bacterial Adhesion on Titanium

The objective of this study was to investigate the potential of titanium nanotubes to promote the proliferation of human osteoblasts and to reduce monomicrobial biofilm adhesion. A secondary objective was to determine the effect of silicon carbide (SiC) on these nanostructured surfaces. Anodized titanium sheets with 100–150 nm nanotubes were either coated or not coated with SiC. After 24 h of osteoblast cultivation on the samples, cells were observed on all titanium sheets by SEM. In addition, the cytotoxicity was evaluated by CellTiter-BlueCell assay after 1, 3, and 7 days. The samples were also cultivated in culture medium with microorganisms incubated anaerobically with respective predominant periodontal bacteria viz. Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia as monoinfection at 37 °C for 30 days. The biofilm adhesion and coverage were evaluated through surface observation using Scanning Electron Microscopy (SEM). The results demonstrate that Ti nanostructured surfaces induced more cell proliferation after seven days. All groups presented no cytotoxic effects on human osteoblasts. In addition, SEM images illustrate that Ti nanostructured surfaces exhibited lower biofilm coverage compared to the reference samples. These results indicate that Ti nanotubes promoted osteoblasts proliferation and induced cell proliferation on the surface, compared with the controls. Ti nanotubes also reduced biofilm adhesion on titanium implant surfaces.     

Integrin alpha2beta1 plays a critical role in osteoblast response to micron-scale surface structure and surface energy of titanium substrates

Efforts to improve bone response to biomaterials have focused on ligands that bind α5β1 integrins. However, antibodies to α5β1 reduce osteoblast proliferation but do not affect differentiation when cells are grown on titanium (Ti). β1-silencing blocks the differentiation stimulus of Ti microtopography, suggesting that other β1 partners are important. Stably α2-silenced MG63 human osteoblast-like cells were used to test whether α2β1 specifically mediates osteoblast response to Ti surface micron-scale structure and energy. WT and α2-silenced MG63 cells were cultured on tissue culture polystyrene (TCPS) and Ti disks with different surface microtopographies: machined pretreatment (PT) surfaces [mean peak to valley roughness (R_a) < 0.02 μm], PT surfaces that were grit-blasted and acid-etched (SLA; R_a = 4 μm), and SLA with high surface energy (modSLA). Alkaline phosphatase (ALP), α2 and β1 mRNA, but not α5, αv, β3, type-I collagen, or osteocalcin, increased on SLA and modSLA at 6 days. α2 increased at 8 days on TCPS and PT, but remained unchanged on SLA and modSLA. α2-protein was reduced 70% in α2-siRNA cells, whereas α5-mRNA and protein were unaffected. α2-knockdown blocked surface-dependent increases in β1 and osteocalcin and decreases in cell number and increases in ALP and local factors typical of MG63 cells grown on SLA and modSLA [e.g., prostaglandin E₂, osteoprotegerin, latent and active TGF-β1, and stimulatory effects of 1α,25(OH)₂D₃ on these parameters]. This finding indicates that α2β1 signaling is required for osteoblastic differentiation caused by Ti microstructure and surface energy, suggesting that conclusions based on cell behavior on TCPS are not predictive of behavior on other substrates or the mechanisms involved.     

Effect of a Nanostructured Titanium Surface on Gingival Cell Adhesion,Viability and Properties against P. gingivalis

Objectives: The transgingival part of titanium implants is either machined or polished. Cell-surface interactions as a result of nano-modified surfaces could help gingival fibroblast adhesion and support antibacterial properties by means of the physico-mechanical aspects of the surfaces. The aim of the present study was to determine how a nanocavity titanium surface affects the viability and adhesion of human gingival fibroblasts (HGF-1). Additionally, its properties against Porphyromonas gingivalis were tested. Material and Methods: Two different specimens were evaluated: commercially available machined titanium discs (MD) and nanostructured discs (ND). To obtain ND, machined titanium discs with a diameter of 15 mm were etched with a 1:1 mixture of 98% H₂SO₄ and 30% H₂O₂ (piranha etching) for 5 h at room temperature. Surface topography characterization was performed via scanning electron microscopy (SEM) and atomic force microscopy (AFM). Samples were exposed to HGF-1 to assess the effect on cell viability and adhesion, which were compared between the two groups by means of MTT assay, immunofluorescence and flow cytometry. After incubation with P. gingivalis, antibacterial properties of MD and ND were determined by conventional culturing, live/dead staining and SEM. Results: The present study successfully created a nanostructured surface on commercially available machined titanium discs. The etching process created cavities with a 10–20 nm edge-to-edge diameter. MD and ND show similar adhesion forces equal to about 10–30 nN. The achieved nanostructuration reduced the cell alignment along machining structures and did not negatively affect the proliferation of gingival fibroblasts when compared to MD. No differences in the expression levels of both actin and vinculin proteins, after incubation on MD or ND, were observed. However, the novel ND surface failed to show antibacterial effects against P. gingivalis. Conclusion: Antibacterial effects against P. gingivalis cannot be achieved with nanocavities within a range of 10–20 nm and based on the piranha etching procedure. The proliferation of HGF-1 and the expression levels and localization of the structural proteins actin and vinculin were not influenced by the surface nanostructuration. Further studies on the strength of the gingival cell adhesion should be performed in the future. Clinical relevance: Since osseointegration is well investigated, mucointegration is an important part of future research and developments. Little is known about how nanostructures on the machined transgingival part of an implant could possibly influence the surrounding tissue. Targeting titanium surfaces with improved antimicrobial properties requires extensive preclinical basic research to gain clinical relevance.     

Disinfection and Biocompatibility of Titanium Surfaces Treated with Glycine Powder Airflow and Triple Antibiotic Mixture: An In Vitro Study

The aim of this in vitro study was to compare three disinfection protocols of biofilm-coated machined (MAC) and acid etched (SLA) commercial pure Grade 4 Titanium disks. Samples were infected with a vial of polymicrobial biofilm to simulate peri-implantitis in vitro. Seventeen MAC and twenty SLA titanium disks were randomly assigned to: (1) glycine powder air-flow (GYPAP) for 1 min; (2) a local delivered triple paste antibiotic composed by a gel mixture with ciprofloxacin, metronidazole, and clarithromycin (3MIX) for 1 h; and (3) a combination of both (GYPAP + 3MIX). Biocompatibility of the titanium disks after each treatment protocol was assessed by measurement of adhesion and growth of adipose-derived mesenchymal stem cells (ASCs) after 24 and 72 h. A confocal laser scanning microscope (CLSM) assessed the antibacterial effect of each treatment. Data of the antibacterial efficacy and cell viability were presented as mean with standard deviation and calculated by one-way ANOVA with multiple comparisons via Bonferroni tests. Results were considered significant with p < 0.05. The higher cell viability was achieved by the 3MIX and GYPAP combination on the SLA surfaces after 72 h. CLSM analysis showed a mean ratio of dead bacteria statistically higher in the 3MIX + GYPAP group compared with the GYPAP and 3MIX subgroups (p < 0.05). In conclusion, data showed that the combination of GYPAP and 3MIX could be preferred to the other protocols, especially in presence of SLA titanium surface.     

Mapping Bone Marrow Cell Response from Senile Female Rats on Ca-P-Doped Titanium Coating

Chemical and topographical surface modifications on dental implants aim to increase the bone surface contact area of the implant and improve osseointegration. This study analyzed the cellular response of undifferentiated mesenchymal stem cells (MSC), derived from senile rats’ femoral bone marrow, when cultured on a bioactive coating (by plasma electrolytic oxidation, PEO, with Ca²⁺ and P⁵⁺ ions), a sandblasting followed by acid-etching (SLA) surface, and a machined surface (MSU). A total of 102 Ti-6Al-4V discs were divided into three groups (n = 34). The surface chemistry was analyzed by energy dispersive spectroscopy (EDS). Cell viability assay, gene expression of osteoblastic markers, and mineralized matrix formation were investigated. The cell growth and viability results were higher for PEO vs. MSU surface (p = 0.001). An increase in cell proliferation from 3 to 7 days (p < 0.05) and from 7 to 10 days (p < 0.05) was noted for PEO and SLA surfaces. Gene expression for OSX, ALP, BSP, and OPN showed a statistical significance (p = 0.001) among groups. In addition, the PEO surface showed a higher mineralized matrix bone formation (p = 0.003). In conclusion, MSC from senile female rats cultured on SLA and PEO surfaces showed similar cellular responses and should be considered for future clinical investigations.     

Role of Titanium Surface Topography and Surface Wettability on Focal Adhesion Kinase Mediated Signaling in Fibroblasts

Changes of titanium surface roughness and surface free energy may influence protein absorption that increases cell differentiation through activation of focal adhesion kinase related pathways. However, the influence of titanium surface roughness and hydrophilicity on fibroblast behavior is not well understood. The aim of this study was to investigate the influence of topography and hydrophilicity on fibroblast attachment, spreading, morphology, intracellular signaling, proliferation, and collagen I mRNA levels. Using a cellular FAK knockout (FAK^−/−) model and wild-type (WT) controls, we also investigated the contribution of adhesion in fibroblasts cultured on smooth (PT), sand-blasted, large grit, acid-etched (SLA) and hydrophilic SLA topographies. Loss of FAK did not significantly affect fibroblast attachment to any surface, but SLA and hydrophilic SLA surface attenuated spreading of WT cells significantly more than FAK^−/− fibroblasts. Both FAK^−/− and WT cells formed numerous focal adhesions on PT surfaces, but significantly less on SLA and hydrophilic SLA surfaces. In WT cells, phosphorylation levels of FAK were lower on SLA and hydrophilic SLA in comparison with PT 24 h post seeding. Labeling of cells with antibodies to cortactin showed that FAK^−/− cells contained significantly more cortactin-rich focal adhesion in comparison with WT cells on PT surfaces, but not on SLA or hydrophilic SLA. ERK 1/2 phosphorylation was highest in WT cells on all surfaces which correlated with collagen I expression levels. We conclude that fibroblasts are sensitive to changes in surface roughness and hydrophilicity, with adhesive interactions mediated through FAK, an important modulator of fibroblast response.     

Structurally diverse natural products that cause potassium leakage trigger multicellularity in Bacillus subtilis

We report a previously undescribed quorum-sensing mechanism for triggering multicellularity in Bacillus subtilis. B. subtilis forms communities of cells known as biofilms in response to an unknown signal. We discovered that biofilm formation is stimulated by a variety of small molecules produced by bacteria—including the B. subtilis nonribosomal peptide surfactin—that share the ability to induce potassium leakage. Natural products that do not cause potassium leakage failed to induce multicellularity. Small-molecule-induced multicellularity was prevented by the addition of potassium, but not sodium or lithium. Evidence is presented that potassium leakage stimulates the activity of a membrane protein kinase, KinC, which governs the expression of genes involved in biofilm formation. We propose that KinC responds to lowered intracellular potassium concentration and that this is a quorum-sensing mechanism that enables B. subtilis to respond to related and unrelated bacteria.     

Evaluation of antimicrobial peptide LL-37 for treatment of Staphylococcus aureus biofilm on titanium plate

The antimicrobial peptide LL-37 belongs to the cathelicidin family and is one of the few human bactericidal peptides with potent antistaphylococcal activity. Staphylococcus aureus is one of the main infection bacteria in orthopedic implant therapy. Biofilm formation after bacterial infection brings more and more severe test for clinical antiinfection treatment.However, there are few studies on LL-37 in S. aureus infection of prosthesis. In this work, addition to research the antibacterial activity and the inhibitory effect on bacterial adhesion of LL-37, an in vitro model of S. aureus biofilm formation on titanium alloy surface was established to observe the inhibitory effect of LL-37.The results showed that LL-37 has a strong antibacterial effect on S. aureus in vitro, and the minimum inhibitory concentration (MIC) is about 0.62 μΜ. Moreover, LL-37 has significant impact on the adhesion of S. aureus when the concentration ≥0.16 μM and significant anti-staphylococcal biofilm effects on static biofilm models at the concentration of 0.31 to 10 μM. Additionally, LL-37 at 5 μM had a significant destructive effect on S. aureus biofilm (P < .05) that formed on the titanium alloy surface.This study further confirmed the role of LL-37 in the process of S. aureus infection, including antimicrobial activities, inhibition of bacterial adhesion, and inhibition of mature biofilm. LL-37 can significantly destroy the stable biofilm structure on the titanium alloy surface in vitro, which may provide a new way for refractory infection caused by S. aureus in titanium alloy prosthesis infection.     

Corrosion Resistance and Surface Bioactivity of Ti6Al4V Alloy after Finish Turning under Ecological Cutting Conditions

The influence of cooling conditions and surface topography after finish turning of Ti6Al4V titanium alloy on corrosion resistance and surface bioactivity was analyzed. The samples were machined under dry and minimum quantity lubrication (MQL) conditions to obtain different surface roughness. The surface topographies of the processed samples were assessed and measured using an optical profilometer. The produced samples were subjected to electrochemical impedance spectroscopy (EIS) and corrosion potential tests (E_corr) in the presence of simulated body fluid (SBF). The surface bioactivity of the samples was assessed on the basis of images from scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS) analysis. The inspection of the surfaces of samples after turning under dry and MQL conditions revealed unevenly distributed precipitation of hydroxyapatite compounds (Ca/P) with a molar ratio in the range of 1.73–1.97. Regardless of the cutting conditions and surface roughness, the highest values of E_corr ~0 mV were recorded on day 7 of immersion in the SBF solution. The impedance characteristics showed that, compared to the MQL conditions, surfaces machined under dry conditions were characterized by greater resistance and the presence of a passive layer on the processed surface. The main novelty of the paper is the study of the effect of ecological machining conditions, namely, dry and MQL cutting on the corrosion resistance and surface bioactivity of Ti6Al4V titanium alloy after finish turning. The obtained research results have practical significance. They can be used by engineers during the development of technological processes for medical devices made of Ti6Al4V alloy to obtain favorable functional properties of these devices.     

Characterization of Hydroxyapatite Film Obtained by Er:YAG Pulsed Laser Deposition on Sandblasted Titanium: An In Vitro Study

The surface of titanium (Ti) dental implants must be modified to improve their applicability, owing to the biological inertness of Ti. This study aims to use sandblasting as a pretreatment method and prepare a hydroxyapatite (HA) coating on Ti to improve its biocompatibility and induce bone bonding and osteogenesis. In this paper, sandblasted Ti discs were coated with α-tricalcium phosphate (α-TCP) via Er:YAG pulsed laser deposition (Er:YAG-PLD). An HA coating was then obtained via the hydrothermal treatment of the discs at 90 °C for 10 h. The surface characteristics of the samples were evaluated by SEM, SPM, XPS, XRD, FTIR, and tensile tests. Rat bone marrow mesenchymal stem cells were seeded on the HA-coated discs to determine cellular responses in vitro. The surface characterization results indicated the successful transformation of the HA coating with a nanorod-like morphology, and its surface roughness increased. In vitro experiments revealed increased cell attachment on the HA-coated discs, as did the cell morphology of fluorescence staining and SEM analysis; in contrast, there was no increase in cell proliferation. This study confirms that Er:YAG-PLD could be used as an implant surface-modification technique to prepare HA coatings with a nanorod-like morphology on Ti discs.     

Morphogenesis and cell ordering in confined bacterial biofilms

Biofilms are aggregates of bacterial cells surrounded by an extracellular matrix. Much progress has been made in studying biofilm growth on solid substrates; however, little is known about the biophysical mechanisms underlying biofilm development in three-dimensional confined environments in which the biofilm-dwelling cells must push against and even damage the surrounding environment to proliferate. Here, combining single-cell imaging, mutagenesis, and rheological measurement, we reveal the key morphogenesis steps of Vibrio cholerae biofilms embedded in hydrogels as they grow by four orders of magnitude from their initial size. We show that the morphodynamics and cell ordering in embedded biofilms are fundamentally different from those of biofilms on flat surfaces. Treating embedded biofilms as inclusions growing in an elastic medium, we quantitatively show that the stiffness contrast between the biofilm and its environment determines biofilm morphology and internal architecture, selecting between spherical biofilms with no cell ordering and oblate ellipsoidal biofilms with high cell ordering. When embedded in stiff gels, cells self-organize into a bipolar structure that resembles the molecular ordering in nematic liquid crystal droplets. In vitro biomechanical analysis shows that cell ordering arises from stress transmission across the biofilm–environment interface, mediated by specific matrix components. Our imaging technique and theoretical approach are generalizable to other biofilm-forming species and potentially to biofilms embedded in mucus or host tissues as during infection. Our results open an avenue to understand how confined cell communities grow by means of a compromise between their inherent developmental program and the mechanical constraints imposed by the environment.

The growth of living organisms is critically influenced by the external environment. One form of such environmental influence is the transmission of mechanical stress, which can instruct morphogenesis in systems ranging from stem cells to tissues to the entire organisms (1, 2). In the prokaryotic domain, bacteria commonly live in complex communities encased by an extracellular matrix (3) known as biofilms (4). Biofilm formation is a morphogenetic process whereby a single founder cell develops into a three-dimensional (3D) aggregate in which bacterial cells interact with each other and with the environment (4–8). Recent work has revealed biophysical mechanisms underlying biofilm morphogenesis on solid substrates, which is controlled by cell–substrate adhesion and the resulting shear stress (9–15). In addition to those living on surfaces, bacterial communities are also commonly found inside soft, structured environments, such as hydrogels. Examples include biofilms growing in mucus layers and host tissues during an infection or food contamination (16). Indeed, many common biofilm formers including Pseudomonas aeruginosa and Vibrio cholerae encounter biological hydrogels as their niche during infection (17, 18). Under these conditions, embedded biofilms must grow against 3D confinement and potentially damage the surrounding environment—a process that is fundamentally different from how surface-attached biofilms expand against friction with the surface (10, 13, 15, 19). However, little is known about how biofilms develop under such 3D mechanical constraints, including how cells collectively organize in response to the confinement and how the confining environment, in turn, is modified by cell proliferation. This is in stark contrast to the accumulating knowledge on the growth dynamics of mammalian cell aggregates and tumors under confinement (20, 21).In this study, we integrate single-cell live imaging, mutagenesis, in vitro mechanical testing, and numerical modeling to investigate how the 3D confinement determines the morphodynamics and cell ordering of an embedded biofilm. A model system is established by embedding V. cholerae, the causal agent of the cholera pandemic and a model biofilm former (22, 23), inside agarose gels (24). By using 3D visualization techniques with high spatiotemporal resolution, we reveal that embedded biofilms undergo a shape transition and a series of self-organization events that are distinct from those in surface-attached biofilms. We first show that the stiffness contrast between the biofilm and the confining hydrogels controls a transition between spherical and ellipsoidal biofilms. Furthermore, we discover that embedded biofilms display a core-shell structure with intricate ordering similar to nematic liquid crystal (LC) droplets (25). Finally, we demonstrate that Vibrio polysaccharide (VPS) and cell-to-surface adhesion proteins effectively transmit stress between the environment and the biofilm, giving rise to distinct cell ordering patterns in embedded biofilms.     

Assessment of Disinfection Potential of Q-Switch Nd: YAG Laser on Contaminated Titanium Implant Surfaces

Peri-implantitis (PI) is a relatively frequent pathology that compromises the overall survival of the dental implant. Adjunctive approaches for the conventional mechanical debridement are being suggested to optimize the treatment of PI. The goal of the study was the assessment of the disinfection potential of the Q-Switch Nd: YAG laser on contaminated titanium implant surfaces. A total of 72 sterile titanium discs were used and divided into three groups: 24 contaminated titanium discs treated with the laser (study Group L), 24 contaminated titanium discs with no treatment (control 1—Group C), and 24 sterile titanium discs with no treatment (control 2—Group S). Multi-species biofilm was used: Porphyromonas gingivalis, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Streptococcus mutans, Streptococcus sobrinus, and Prevotella intermedia. Commensal bacteria were included also: Actinomyces naeslundii, Actinomyces viscosus, Streptococcus cristatus, Streptococcus gordonii, Streptococcus mitis, Streptococcus oralis, Streptococcus sanguinis, Streptococcus parasanguinis, and Veillonella parvula. Parameters delivered per pulse on the targeted surfaces of the titanium discs were an energy density of 0.597 J/cm² each pulse, a pulse power of 270 mW, a laser beam spot of 2.4 mm in diameter, and a rate of repetition of 10 Hertz (Hz) for a pulse duration of 6 nanoseconds (ns). The mode was no contact, and a distance of 500 micrometers was used with a total time of irradiation equal to 2 s (s). The collection of microbiological samples was made for all groups; colony-forming units (CFU) were identified by two different practitioners, and the average of their examinations was considered for each sample. The average of the TBC (CFU/mL) was calculated for each group. Values were 0.000 CFU/mL, 4767 CFU/mL, and 0.000 CFU/mL for Group L, Group C, and Group S, respectively. Therefore, the suggested treatment protocol was able to provoke a total disinfection of the contaminated titanium surfaces. A statistical difference was only found between Group L vs. Group C and between Group S vs. Group C. The difference was not significant between Group S and Group L. In conclusion, the present study confirmed that the Q-Switch Nd: YAG laser under our specific conditions can provide a total disinfection of the contaminated titanium surfaces.     

DNA Polyelectrolyte Multilayer Coatings Are Antifouling and Promote Mammalian Cell Adhesion

The ability of bacteria to adhere to and form biofilms on implant surfaces is the primary cause of implant failure. Implant-associated infections are difficult to treat, as the biofilm mode of growth protects microorganisms from the host’s immune response and antibiotics. Therefore, modifications of implant surfaces that can prevent or delay bacterial adhesion and biofilm formation are highly desired. In addition, the attachment and spreading of bone cells are required for successful tissue integration in orthopedic and dental applications. We propose that polyanionic DNA with a negatively charged phosphate backbone could provide a dual function to repel bacterial adhesion and support host tissue cell attachment. To this end, we developed polyelectrolyte multilayer coatings using chitosan (CS) and DNA on biomaterial surfaces via a layer-by-layer technique. The assembly of these coatings was characterized. Further, we evaluated staphylococcal adhesion and biofilm growth on the coatings as well as cytotoxicity for osteoblast-like cells (SaOS-2 cells), and we correlated these to the layer structure. The CS-DNA multilayer coatings impaired the biofilm formation of Staphylococcus by ~90% on both PMMA and titanium surfaces. The presence of cationic CS as the top layer did not hinder the bacteria-repelling property of the DNA in the coating. The CS-DNA multilayer coatings demonstrated no cytotoxic effect on SaOS-2 cells. Thus, DNA polyelectrolyte multilayer coatings could reduce infection risk while promoting host tissue cell attachment on medical implants.     

Human Gingival Fibroblast and Osteoblast Behavior on Groove-Milled Zirconia Implant Surfaces

Two type of cells representing periodontal hard tissues (osteoblasts) and soft tissues (fibroblasts) were evaluated in response to microgroove-milled zirconia surfaces. A total of 90 zirconia discs were randomly assigned to four width-standardized milling microgroove-textured groups and a control group without grooves (UT). The sandblast and acid-etch protocol were applied to all samples. Both cell lines were cultured on zirconia discs from 1 day up to 14 days. Cell morphology and adhesion were evaluated after 1 day of culturing. Cell viability and proliferation of the cells were measured. Alkaline phosphatase activity, collagen I, osteopontin, interleukin 1β and interleukin 8 secretions were assessed at predefined times. The results obtained were presented in the form of bar graphs as means and standard deviations. Multi comparisons between groups were evaluated using two-away ANOVA or Mann–Whitney tests, and a p-value < 0.05 was established. Group comparisons with regard to cell viability, proliferation and secretion of collagen I, interleukin-1β and interleukin 8 revealed no statistically significant differences. The alkaline phosphatase activity and osteopontin secretion were significantly higher in the group with a large groove compared to the small one and the control group. Nevertheless, the viability of gingival and bone cells did not appear to be affected by the milled microgroove texture compared to the conventional sandblasted and acid-etched texture, but they seem to influence osteoblasts’ cellular differentiation.     

Effects of Gamma Radiation-Induced Crosslinking of Collagen Type I Coated Dental Titanium Implants on Osseointegration and Bone Regeneration

This study aimed to compare two methods of crosslinking collagen type I on implanted titanium surfaces, that is, using glutaraldehyde (GA) or gamma-rays (GRs), in a beagle dog model. For in vivo experiments, implants were allocated to three groups and applied to mandibular bone defects in beagle dogs; Group SLA; non-treated Sandblasted, large grit, acid-etched (SLA) implants, Group GA; SLA implants coated with GA crosslinked collagen type I, Group GR; SLA surface implants coated with collagen type I and crosslinked using 25 kGy of ⁶⁰Co gamma radiation. New bone μCT volumes were obtained, and histologic and histometric analyses were performed in regions of interest. The GR group had significantly better new bone areas (NBAs) and bone to implant contact (BIC) results than the SLA group (p < 0.05), but the GA and GR groups were similar in this respect. New bone volumes and inter-thread bone densities (ITBD) were non-significantly different in the three groups (p > 0.05). Within the limits of this study, gamma-ray collagen crosslinking on titanium implants can be considered a substitute for glutaraldehyde crosslinking.     

Greater Osseointegration Potential with Nanostructured Surfaces on TiZr: Accelerated vs. Real-Time Ageing

Surface chemistry and nanotopography of dental implants can have a substantial impact on osseointegration. The aim of this investigation was to evaluate the effects of surface chemistry and nanotopography on the osseointegration of titanium-zirconium (TiZr; Roxolid^®) discs, using a biomechanical pull-out model in rabbits. Two discs each were placed in both the right and left tibiae of 16 rabbits. Five groups of sandblasted acid etched (SLA) discs were tested: (1) hydrophobic without nanostructures (dry/micro) (n = 13); (2) hydrophobic with nanostructures, accelerated aged (dry/nano/AA) (n = 12); (3) hydrophilic without nanostructures (wet/micro) (n = 13); (4) hydrophilic with nanostructures, accelerated aged (wet/nano/AA; SLActive^®) (n = 13); (5) hydrophilic with nanostructures, real-time aged (wet/nano/RTA). The animals were sacrificed after four weeks and the biomechanical pull-out force required to remove the discs was evaluated. Adjusted mean pull-out force was greatest for group wet/nano/RTA (64.5 ± 17.7 N) and lowest for group dry/micro (33.8 ± 10.7 N). Multivariate mixed model analysis showed that the pull-out force was significantly greater for all other disc types compared to the dry/micro group. Surface chemistry and topography both had a significant effect on pull-out force (p < 0.0001 for both), but the effect of the interaction between chemistry and topography was not significant (p = 0.1056). The introduction of nanostructures on the TiZr surface significantly increases osseointegration. The introduction of hydrophilicity to the TiZr implant surface significantly increases the capacity for osseointegration, irrespective of the presence or absence of nanotopography.     

Efficacy of Surgical/Wound Washes against Bacteria: Effect of Different In Vitro Models

Topical antiseptics are often used to treat chronic wounds with biofilm infections and during salvage of biofilm contaminated implants, but their antibacterial efficacy is frequently only tested against non-aggregated planktonic or free-swimming organisms. This study evaluated the antibacterial and antibiofilm efficacy of four commercial surgical washes Bactisure, TorrenTX, minimally invasive lavage (MIS), and Betadine against six bacterial species: Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pyogenes, Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli, which are commonly isolated from surgical site infections and chronic wound infections using different in vitro models. We determined minimum planktonic inhibitory and eradication concentration and minimum 1-day-old biofilm inhibition and eradication concentration of antiseptics in 96-well plates format with 24 h contact time. We also tested the efficacy of antiseptics at in-use concentration and contact time in the presence of biological soil against 3-day-old biofilm grown on coupons with shear in a bioreactor, such that the results are more applicable to the clinical biofilm situations. In the 96-well plate model, the minimum concentration required to inhibit or kill planktonic and biofilm bacteria was lower for Bactisure and TorrenTX than for MIS and Betadine. However, Betadine and Bactisure showed better antibiofilm efficacy than TorrenTX and MIS in the 3-day-old biofilm bioreactor model at in-use concentration. The minimal concentration of surgical washes required to inhibit or kill planktonic bacterial cells and biofilms varies, suggesting the need for the development and use of biofilm-based assays to assess antimicrobial therapies, such as topical antiseptics and their effective concentrations. The antibiofilm efficacy of surgical washes against different bacterial species also varies, highlighting the importance of testing against various bacterial species to achieve a thorough understanding of their efficacy.     

Novel Coatings to Minimize Corrosion of Titanium in Oral Biofilm

The aim of this work is to investigate the effects produced by polymicrobial biofilm (Porphyromonas gingivalis, Streptococcus mutans, Streptococcus sanguinis, and Streptococcus salivarius) on the corrosion behavior of titanium dental implants. Pure titanium disks were polished and coated with titanium nitride (TiN) and silicon carbide (SiC) along with their quarternized versions. Next, the disks were cultivated in culture medium (BHI) with P. gingivalis, S. mutans, S. sanguinis, and S. salivarius and incubated anaerobically at 37 °C for 30 days. Titanium corrosion was evaluated through surface observation using Scanning Electron Microscope (SEM) and Atomic Force Microscopy (AFM). Furthermore, the Ti release in the medium was evaluated by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). SEM images showed that coated Ti disks exhibited lower corrosion compared to non-coated disks, except for the quartenized TiN. This was confirmed by AFM, where the roughness was higher in non-coated Ti disks. ICP showed that Ti levels were low in all coating disks. These results indicate that these SiC and TiN-based coatings could be a useful tool to reduce surface corrosion on titanium implant surfaces.