Increased use of titanium alloys as biomaterials is occurring due to their lower modulus, superior biocompatibility and enhanced corrosion resistance when compared to more conventional stainless steels and cobalt-based alloys. These attractive properties were a driving force for the early introduction of alpha (cpTi) and alpha + beta (Ti-6A1-4V) alloys as well as for the more recent development of new Ti-alloy compositions and orthopaedic metastable beta titanium alloys. The later possess enhanced biocompatibility, reduced elastic modulus, and superior strain-controlled and notch fatigue resistance. However, the poor shear strength and wear resistance of titanium alloys have nevertheless limited their biomedical use. Although the wear resistance of beta-Ti alloys has shown some improvement when compared to alpha + beta alloys, the ultimate utility of orthopaedic titanium alloys as wear components will require a more complete fundamental understanding of the wear mechanisms involved. This review examines current information on the physical and mechanical characteristics of titanium alloys used in artifical joint replacement prostheses, with a special focus on those issues associated with the long-term prosthetic requirements, e.g., fatigue and wear.

Cobalt (Co) is an essential element with ubiquitous dietary exposure and possible incremental exposure due to dietary supplements, occupation and medical devices. Adverse health effects, such as cardiomyopathy and vision or hearing impairment, were reported at peak blood Co concentrations typically over 700 µg/L (8-40 weeks), while reversible hypothyroidism and polycythemia were reported in humans at ~300 µg/L and higher (≥2 weeks). Lung cancer risks associated with certain inhalation exposures have not been observed following Co ingestion and Co alloy implants. The mode of action for systemic toxicity relates directly to free Co(II) ion interactions with various receptors, ion channels and biomolecules resulting in generally reversible effects. Certain dose-response anomalies for Co toxicity likely relate to rare disease states known to reduce systemic Co(II)-ion binding to blood proteins. Based on the available information, most people with clearly elevated serum Co, like supplement users and hip implant patients, have >90% of Co as albumin-bound, with considerable excess binding capacity to sequester Co(II) ions. This paper reviews the scientific literature regarding the chemistry, pharmacokinetics and systemic toxicology of Co, and the likely role of free Co(II) ions to explain dose-response relationships. Based on currently available data, it might be useful to monitor implant patients for signs of hypothyroidism and polycythemia starting at blood or serum Co concentrations above 100 µg/L. This concentration is derived by applying an uncertainty factor of 3 to the 300 µg/L point of departure and this should adequately account for the fact that persons in the various studies were exposed for less than one year. A higher uncertainty factor could be warranted but Co has a relatively fast elimination, and many of the populations studied were of children and those with kidney problems. Closer follow-up of patients who also exhibit chronic disease states leading to clinically important hypoalbuminemia and/or severe ischemia modified albumin (IMA) elevations should be considered.

Biomedical titanium alloys with Young's moduli close to that of cortical bone, i.e., low Young's modulus titanium alloys, are receiving extensive attentions because of their potential in preventing stress shielding, which usually leads to bone resorption and poor bone remodeling, when implants made of their alloys are used. They are generally β-type titanium alloys composed of non-toxic and allergy-free elements such as Ti-29Nb-13Ta-4.6Zr referred to as TNTZ, which is highly expected to be used as a biomaterial for implants replacing failed hard tissue. Furthermore, to satisfy the demands from both patients and surgeons, i.e., a low Young's modulus of the whole implant and a high Young's modulus of the deformed part of implant, titanium alloys with changeable Young's modulus, which are also β-type titanium alloys, for instance Ti-12Cr, have been developed. In this review article, by focusing on TNTZ and Ti-12Cr, the biological and mechanical properties of the titanium alloys with low Young's modulus and changeable Young's modulus are described. In addition, the titanium alloys with shape memory and superelastic properties were briefly addressed. Surface modifications for tailoring the biological and anti-wear/corrosion performances of the alloys have also been briefly introduced.

Two-piece implant systems are mainly used in oral implantology involving an osseointegrated implant connected to an abutment, which supports prosthetic structures. It is well documented that the presence of microgaps, biofilms and oral fluids at the implant-abutment connection can cause mechanical and biological complications. The aim of this review paper was to report the degradation at the implant-abutment connection by wear and corrosion processes taking place in the oral cavity. Most of the retrieved studies evaluated the wear and corrosion (tribocorrosion) of titanium-based materials used for implants and abutments in artificial saliva. Electrochemical and wear tests together with microscopic techniques were applied to validate the tribocorrosion behavior of the surfaces. A few studies inspected the wear on the inner surfaces of the implant connection as a result of fatigue or removal of abutments. The studies reported increased microgaps after fatigue tests. In addition, data suggest that micromovements occurring at the contacting surfaces can increase the wear of the inner surfaces of the connection. Biofilms and/or glycoproteins act as lubricants, although they can also amplify the corrosion of the surfaces. Consequently, loosening of the implant-abutment connection can take place during mastication. In addition, wear and corrosion debris such as ions and micro- and nanoparticles released into the surrounding tissues can stimulate peri-implant inflammation that can lead to pathologic bone resorption.© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Inflammation is an important step in the early phase of tissue regeneration around an implanted metallic orthopaedic device. However, prolonged inflammation, which can be induced by metallic corrosion products, can lead to aseptic loosening and implant failure. Cells in peri-implant tissue as well as metal corrosion can induce reactive oxygen species (ROS) formation, thus contributing to an oxidative microenvironment around an implant. Understanding cellular reactions to implant-induced oxidative stress and inflammatory activation is important to help prevent an adverse response to metallic materials. In an earlier study we have shown that endothelial cells grown on Ti6Al4V alloy are subjected to oxidative stress. Since endothelial cells play a critical role in inflammation, in this study we examined the role of oxidative stress in their response to pro-inflammatory activation. Therefore, we stimulated endothelial cells in contact with Ti6Al4V with tumour necrosis factor-α (TNF-α) and monitored the expression of inflammation-associated molecules, such as E-selectin, intercellular adhesion molecule-1 (ICAM-1) and interleukin-8 (IL-8). The induction of these proteins was lower in endothelial cells on Ti6Al4V compared to control tissue culture conditions. There was, however, a discrepancy in pro-inflammatory activation at protein compared to mRNA level in the cells on Ti6Al4V. To examine the role of oxidative stress in this response we utilized different ROS scavengers and showed that ROS depletion improved cellular response to TNF-α on Ti6Al4V. These results could contribute to developing strategies to improve tissue response to metal implants. © 2013 Elsevier Ltd. All rights reserved.

The fact that aluminium (Al) and lead (Pb) are both toxic metals to living organisms, including human beings, was discovered a long time ago. Even when Al and Pb can reach and accumulate in almost every organ in the human body, the central nervous system is a particular target of the deleterious effects of both metals. Select human population can be at risk of Al neurotoxicity, and Al is proposed to be involved in the etiology of neurodegenerative diseases. Pb is a widespread environmental hazard, and the neurotoxic effects of Pb are a major public health concern. In spite of the numerous efforts and the accumulating evidence in this area of research, the mechanisms of Al and Pb neurotoxicity are still not completely elucidated. This review will particularly address the involvement of oxidative stress, membrane biophysics alterations, deregulation of cell signaling, and the impairment of neurotransmission as key aspects involved Al and Pb neurotoxicity.

Implanted biomaterials play a key role in the current success of orthopedic and dental procedures. Pure titanium and its alloys are the most commonly used materials for permanent implants in contact with bone. However, implant-related infections remain among the leading reasons for failure. The most critical pathogenic event in the development of infection on biomaterials is biofilm formation, which starts immediately after bacterial adhesion. In the last decade, numerous studies reported the ability of titanium surface modifications and coatings to minimize bacterial adhesion, inhibit biofilm formation and provide effective bacterial killing to protect implanted biomaterials. In the present review, the different strategies to prevent infection onto titanium surfaces are reported: surface modification and coatings by antibiotics, antimicrobial peptides, inorganic antibacterial metal elements and antibacterial polymers. STATEMENT OF SIGNIFICANCE: Implanted biomaterials play a key role in the current success of orthopedic and dental procedures. Pure titanium and its alloys are the most commonly used materials for permanent implants in contact with bone. Microbial infection is one of the main causes of implant failure. Currently, the global infection risk is 2-5% in orthopedic surgery. Numerous solutions exist to render titanium surfaces antibacterial. The LBPS team is an expert on the functionalization of titanium surfaces by using bioactive polymers to improve the biologiocal response. In this review, the different strategies to prevent infection are reported onto titanium and titanium alloy surfaces such as surface modification by antibiotics, antimicrobial peptides, inorganic antibacterial metal elements and antibacterial polymers.Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

In this study, the microstructure, mechanical properties, castability, electrochemical behaviors, cytotoxicity and hemocompatibility of Ti-Bi alloys with pure Ti as control were systematically investigated to assess their potential applications in the dental field. The experimental results showed that, except for the Ti-20Bi alloy, the microstructure of all other Ti-Bi alloys exhibit single α-Ti phase, while Ti-20Bi alloy is consisted of mainly α-Ti phase and a small amount of BiTi2 and BiTi3 phases. The tensile strength, hardness and wear resistance of Ti-Bi alloys were demonstrated to be improved monotonically with the increase of Bi content. The castability test showed that Ti-2Bi alloy increased the castability of pure Ti by 11.7%. The studied Ti-Bi alloys showed better corrosion resistance than pure Ti in both AS (artificial saliva) and ASFL (AS containing 0.2% NaF and 0.3% lactic acid) solutions. The concentrations of both Ti ion and Bi ion released from Ti-Bi alloys are extremely low in AS, ASF (AS containing 0.2% NaF) and ASL (AS containing 0.3% lactic acid) solutions. However, in ASFL solution, a large number of Ti and Bi ions are released. In addition, Ti-Bi alloys produced no significant deleterious effect to L929 cells and MG63 cells, similar to pure Ti, indicating a good in vitro biocompatibility. Besides, both L929 and MG63 cells perform excellent cell adhesion ability on Ti-Bi alloys. The hemolysis test exhibited that Ti-Bi alloys have an ultra-low hemolysis percentage below 1% and are considered nonhemolytic. To sum up, the Ti-2Bi alloy exhibits the optimal comprehensive performance and has great potential for dental applications. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Low modulus β-titanium alloys with non-toxic alloying elements are envisaged to provide good biocompatibility and alleviate the undesired stress shielding effect. The objective of this study is to fundamentally elucidate the biological response of novel high strength-low elastic modulus Ti2448 alloy through the study of bioactivity and osteoblast cell functions.Characterization techniques such as SEM, EDX, XRD, and fluorescence microscopy were utilized to analyze the microstructure, morphology, chemical composition, and cell adhesion. The cellular activity was explored in terms of cell-to-cell communication involving proliferation, spreading, synthesis of extracellular and intracellular proteins, differentiation, and mineralization.The formation of fine apatite-like crystals on the surface during immersion test in simulated body fluid confirmed the bioactivity of the surface, which provided the favorable osteogenic microenvironment for cell-material interaction. The proliferation and differentiation of pre-osteoblasts and their ability to form a well mineralized bone-like extracellular matrix (ECM) by secreting bone markers (ALP, calcium, etc.) over the surface point toward the determining role of unique surface chemistry and surface properties of the Ti-24Nb-4Zr-8Sn (Ti2448) alloy in modulating osteoblasts functions.These results demonstrated that the low modulus (∼49GPa) Ti2448 alloy with non-toxic alloying elements can be used as a potential dental or orthopedic load-bearing implant material.Copyright © 2016 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

A new kind of biomedical shape memory TiNiAg alloy with antibacterial function was successfully developed in the present study by the introduction of pure Ag precipitates into the TiNi matrix phase. The microstructure, mechanical property, corrosion resistance, ion release behavior in simulated body fluid, cytotoxicity and antibacterial properties were systematically investigated. The typical microstructural feature of TiNiAg alloy at room temperature was tiny pure Ag particles (at submicrometer or micrometer scales with irregular shape) randomly distributed in the TiNi matrix phase. The presence of Ag precipitates was found to result in a slightly higher tensile strength and larger elongation of TiNiAg alloy in comparison with that of TiNi binary alloy. Furthermore, a maximum shape recovery strain of ∼6.4% was obtained with a total prestrain of 7% in the TiNiAg alloy. In electrochemical and immersion tests, TiNiAg alloy presented good corrosion resistance in simulated body fluid, comparable with that of CP Ti and TiNi alloy. The cytotoxicity evaluation revealed that TiNiAg alloy extract induced slight toxicity to cells, but the viability of experimental cells was similar to or higher than that of TiNi alloy extract. In vitro bacterial adhesion study indicated a significantly reduced number of bacteria (S. aureus, S. epidermidis and P. gingivalis) on the TiNiAg alloy plate surface when compared with that on TiNi alloy plate surface, and the corresponding antibacterial mechanism for the TiNiAg alloy is discussed.Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Sandblasting is a procedure of increasing surface roughness. This treatment is common in the orthopedic field. An increased roughness may affect the endurance limit of the material. This study investigates the effect on the endurance limit of the Ti6Al4V due to two different sandblasting treatments: fine sandblasting and coarse sandblasting. Twenty hip stems, 10 finely sandblasted and 10 coarsely sandblasted, were tested under sinusoidal fluctuating bending. The staircase method was used to estimate the endurance limit of the material. The results show an important reduction in the endurance properties up to 40% for the coarsely sandblasted specimens. The failures of the sandblasted specimens were not due to material defects. Rather, the decreased endurance strength of the sandblasted stems was caused by surface defects, which act as crack initiators. By modulating the roughness with an appropriate sandblasting treatment, it is possible to limit the reduction in the endurance limit of the alloy.

Often hard implants undergo detachment from the host tissue due to inadequate biocompatibility and poor osteointegration. Changing surface chemistry and physical topography of the surface influences biocompatibility. At present, the understanding of biocompatibility of both virgin and modified surfaces of bioimplant materials is limited and a great deal of research is being dedicated to this aspect. In view of this, the current review casts new light on research related to the surface modification of biomaterials, especially materials for prosthetic applications. A brief overview of the major surface modification techniques has been presented, followed by an in-depth discussion on laser surface modifications that have been explored so far along with those that hold tremendous potential for bioimplant applications.

Recently, the production of well-defined patterned surfaces with random or regular micro and nano-features has brought new opportunities for research and development in the field of tissue engineering and regenerative medicine. Among advanced micro and nano processing technologies, laser surface texturing (LST) stands out due to its simplicity, flexibility, precision, reproducibility and relatively low cost. This work studies the development of patterned surfaces controlled by of LST into biomedical grade V titanium, Ti-6Al-4V-alloy. We present different cross-hatched micropatterns followed by the characterisation of surface morphology and topography. Structural integrity of the produced patterns is evaluated by friction tests against bone, mimicking the insertion of an implant. Wettability is studied as it is crucial for protein adsorption and cell adhesion. The results show that the surface topography obtained using different patterning plans influences the wetting behaviour and the coefficient of friction against bone.Copyright © 2019 Elsevier Ltd. All rights reserved.

A systematic analysis of results available from in vitro, in vivo and clinical trials on the effects of biocompatible calcium phosphate (CaP) coatings is presented. An overview of the most frequently used methods to prepare CaP-based coatings was conducted. Dense, homogeneous, highly adherent and biocompatible CaP or hybrid organic/inorganic CaP coatings with tailored properties can be deposited. It has been demonstrated that CaP coatings have a significant effect on the bone regeneration process. In vitro experiments using different cells (e.g. SaOS-2, human mesenchymal stem cells and osteoblast-like cells) have revealed that CaP coatings enhance cellular adhesion, proliferation and differentiation to promote bone regeneration. However, in vivo, the exact mechanism of osteogenesis in response to CaP coatings is unclear; indeed, there are conflicting reports of the effectiveness of CaP coatings, with results ranging from highly effective to no significant or even negative effects. This review therefore highlights progress in CaP coatings for orthopaedic implants and discusses the future research and use of these devices. Currently, an exciting area of research is in bioactive hybrid composite CaP-based coatings containing both inorganic (CaP coating) and organic (collagen, bone morphogenetic proteins, arginylglycylaspartic acid etc.) components with the aim of promoting tissue ingrowth and vascularization. Further investigations are necessary to reveal the relative influences of implant design, surgical procedure, and coating characteristics (thickness, structure, topography, porosity, wettability etc.) on the long-term clinical effects of hybrid CaP coatings. In addition to commercially available plasma spraying, other effective routes for the fabrication of hybrid CaP coatings for clinical use still need to be determined and current progress is discussed. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Osteoblasts grown on microstructured Ti surfaces enhance osteointegration by producing local factors that regulate bone formation as well as bone remodeling, including the RANK ligand decoy receptor osteoprotegerin (OPG). The objective of this study was to explore the mechanism by which surface microstructure and surface energy mediate their stimulatory effects on OPG expression. Titanium disks were manufactured to present different surface morphologies: a smooth pretreatment surface (PT, Ra<0.2microm), microstructured sandblasted/acid etched surface (SLA, Ra=3-4microm), and a microstructured Ti plasma-sprayed surface (TPS, Ra=4microm). Human osteoblast-like MG63 cells were cultured on these substrates and the regulation of OPG production by TGF-beta1, PKC, and alpha2beta1 integrin signaling determined. Osteoblasts produced increased amounts of OPG as well as active and latent TGF-beta1 and had increased PKC activity when grown on SLA and TPS. Exogenous TGF-beta1 increased OPG production in a dose-dependent manner on all surfaces, and this was prevented by adding blocking antibody to the TGF-beta type II receptor or by reducing TGF-beta1 binding to the receptor by adding exogenous soluble type II receptor. The PKC inhibitor chelerythrine inhibited the production of OPG in a dose-dependent manner, but only in cultures on SLA and TPS. shRNA knockdown of alpha2 or a double knockdown of alpha2beta1 also reduced OPG, as well as production of TGF-beta1. These results indicate that substrate-dependent OPG production is regulated by TGF-beta1, PKC, and alpha2beta1 and suggest a mechanism by which alpha2beta1 signaling increases PKC, resulting in TGF-beta1 production and TGF-beta1 then acts on its receptor to increase transcription of OPG.

To evaluate first white titanium surfaces developed for improvement of existing clinically used titanium-based implants Ticer.The anodic plasma-electrochemical oxidation in aqueous solutions of sodium hydroxide and calcium dihydrogen phospate was used to prepare three novel anodic conversion layers with white titanium oxide surfaces. The surfaces have been characterized by the means of scanning electron microscopy, surface microanalysis and X-ray diffraction. In vitro studies were conducted on primary human osteoblast cells using novel surfaces (M1-M3) as well as commercially pure titanium (Ti cp), Ticer and SS (subtracted surface). An indirect toxicity test using MTT and SRB assays has been carried out. Furthermore, immunohistochemical analysis of cell proliferation, morphology, and expression of non-collagenous bone matrix proteins (sialoprotein, BSP, and osteocalcin, OC) were performed.The basic morphology of the surfaces shows clusters in a size of 100 μm of knob-like structures. The coatings are composed of rutile and monoclinic sodium titanates. Novel white surfaces (M1-M3) induced proliferation rates, morphological changes and influenced the expression of OC and BSP similarly to Ticer. On the other hand, Ti cp and SS exhibited different in vitro behavior.The novel surfaces expressed similar in vitro behavior as Ticer, successfully used in clinical practice. Furthermore, due to their white color they are also promising from the esthetic point of view. The results described herein open the door toward a new generation of white titanium dental implants.Copyright © 2014 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Dickkopf-related protein 1 (DKK1) is essential to maintain skeletal homeostasis as an inhibitor of Wnt signaling and osteogenic differentiation. The purpose of this study was to investigate the molecular mechanisms underlying the developmental stage-specific regulation of the DKK1 protein level. We performed a series of studies including luciferase reporter assays, micro-RNA microarray, site-specific mutations, and gain- and loss-of-function analyses. We found that the DKK1 protein level was regulated via DKK1 3' UTR by miRNA control, which was restricted to osteoblast-lineage cells. As a result of decreased DKK1 protein level by miR-335-5p, Wnt signaling was enhanced, as indicated by elevated GSK-3β phosphorylation and increased β-catenin transcriptional activity. The effects of miR-335-5p were reversed by anti-miR-335-5p treatment, which downregulated endogenous miR-335-5p. In vivo studies showed high expression levels of miR-335-5p in osteoblasts and hypertrophic chondrocytes of mouse embryos, indicating a pivotal role of miR-335-5p in regulating bone development. In conclusion, miR-335-5p activates Wnt signaling and promotes osteogenic differentiation by downregulating DKK1. This cell- and development-specific regulation is essential and mandatory for the initiation and progression of osteogenic differentiation. miR-335-5p proves to be a potential and useful targeting molecule for promoting bone formation and regeneration.Copyright © 2011 American Society for Bone and Mineral Research.

Implanted biomaterials are coated immediately with host plasma constituents, including extracellular matrix (ECM); this reaction may be undesirable in some cases. Poly(L-lysine)-grafted-poly(ethylene glycol) (PLL-g-PEG) has been shown to spontaneously adsorb from aqueous solution onto metal oxide surfaces, effectively reducing the degree of non-specific adsorption of blood and ECM proteins, and decreasing the adhesion of fibroblastic and osteoblastic cells to the coated surfaces. Cell adhesion through specific peptide-integrin receptors could be restored on surfaces coated with PLL-g-PEG functionalized with peptides of the RGD (Arg-Asp-Gly) type. To date, no study has examined the effect of surface modifications by PLL-g-PEG-based polymers on bacterial adhesion. The ability of Staphylococcus aureus to adhere to the ECM and plasma proteins deposited on biomaterials is a significant factor in the pathogenesis of medical-device-related infections. This study describes methods for visualizing and quantifying the adhesion of S. aureus to smooth and rough (chemically etched) titanium surfaces without and with monomolecular coatings of PLL-g-PEG, PLL-g-PEG/PEG-RGD and PLL-g-PEG/PEG-RDG. The different surfaces were exposed to S. aureus cultures for 1-24h and bacteria surface density was evaluated using scanning electron microscopy and fluorescence microscopy. Coating titanium surfaces with any of the three types of copolymers significantly decreased the adhesion of S. aureus to the surfaces by 89-93% for PLL-g-PEG, and 69% for PLL-g-PEG/PEG-RGD. Therefore, surfaces coated with PLL-g-PEG/PEG-RGD have the ability to attach cells such as fibroblasts and osteoblasts while showing reduced S. aureus adhesion, resulting in a selective biointeraction pattern that may be useful for applications in the area of osteosynthesis, orthopaedic and dental implantology.

Advances in tissue engineering require biofunctional scaffolds that can not only provide cells with structural support, but also interact with cells in a biological manner. To achieve this goal, a frequently used cell adhesion peptide Arg-Gly-Asp (RGD) was covalently incorporated into poly(ethylene glycol) diacrylate (PEODA) hydrogel and its dosage effect (0.025, 1.25 and 2.5 mm) on osteogenesis of marrow stromal cells in a three-dimensional environment was examined. Expression of bone-related markers, osteocalcin (OCN) and Alkaline phosphatase (ALP), increased significantly as the RGD concentration increased. Compared with no RGD, 2.5 mm RGD group showed a 1344% increase in ALP production and a 277% increase in OCN accumulation in the medium. RGD helped MSCs maintain cbfa-1 expression when shifted from a two-dimensional environment to a three-dimensional environment. Soluble RGD was found to completely block the mineralization of marrow stromal cells, as manifested by quantitative calcium assay, phosphorus elemental analysis and Von Kossa staining. In conclusion, we have demonstrated that RGD-conjugated PEODA hydrogel promotes the osteogenesis of MSCs in a dosage-dependent manner, with 2.5 mm being optimal concentration.

Intravascular catheter-associated infections (CAIs), which are normally induced by microbial adhesion and subsequent biofilm formation, are a major cause of morbidity and mortality. Therefore, strategies to prevent CAIs are in great demand. In this study, a series of diblock copolymers of PEG and cationic polycarbonate with various compositions were synthesized by metal-free organocatalytic ring-opening polymerization, and coated onto silicone rubber (a commonly used catheter material) at different concentrations via a reactive polydopamine coating. Static contact angle and X-ray photoelectron spectroscopy measurements proved the successful coating, and quartz crystal microbalance results showed that the coating thickness increased as polymer concentration increased. Methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) isolates - leading causes of intravascular CAIs - were employed to evaluate the antibacterial and antifouling activities of the polymer coatings. Polymer coatings with a hydrophobic component effectively killed planktonic MSSA and MRSA in solution and prevented their fouling on silicone rubber surface. Live/dead cell staining experiments revealed that polymer coatings with the optimal polymer composition possessed significantly higher antifouling activity than PEG coating. In addition, scanning electron microscopic studies showed that the polymer coating inhibited S. aureus biofilm formation over a period of 7 days. Furthermore, the polymer coating caused no significant hemolysis, and there was no blood protein adsorption or platelet adhesion observed. Therefore, PEG-b-cationic polycarbonates with optimal compositions are effective antifouling and antibacterial coatings for the prevention of intravascular CAIs.Copyright © 2012 Elsevier Ltd. All rights reserved.

Black silicon is a synthetic nanomaterial that contains high aspect ratio nanoprotrusions on its surface, produced through a simple reactive-ion etching technique for use in photovoltaic applications. Surfaces with high aspect-ratio nanofeatures are also common in the natural world, for example, the wings of the dragonfly Diplacodes bipunctata. Here we show that the nanoprotrusions on the surfaces of both black silicon and D. bipunctata wings form hierarchical structures through the formation of clusters of adjacent nanoprotrusions. These structures generate a mechanical bactericidal effect, independent of chemical composition. Both surfaces are highly bactericidal against all tested Gram-negative and Gram-positive bacteria, and endospores, and exhibit estimated average killing rates of up to similar to 450,000 cells min(-1) cm(-2). This represents the first reported physical bactericidal activity of black silicon or indeed for any hydrophilic surface. This biomimetic analogue represents an excellent prospect for the development of a new generation of mechano-responsive, antibacterial nanomaterials.

While insect wings are widely recognised as multi-functional, recent work showed that this extends to extensive bactericidal activity brought about by cell deformation and lysis on the wing nanotopology. We now quantitatively show that subtle changes to this topography result in substantial changes in bactericidal activity that are able to span an order of magnitude. Notably, the chemical composition of the lipid nanopillars was seen by XPS and synchrotron FTIR microspectroscopy to be similar across these activity differences. Modelling the interaction between bacterial cells and the wing surface lipids of 3 species of dragonflies, that inhabit similar environments, but with distinctly different behavioural repertoires, provided the relationship between surface structure and antibacterial functionality. In doing so, these principal behavioural patterns correlated with the demands for antimicrobial efficiency dictated by differences in their foraging strategies. This work now reveals a new feature in the design elegance of natural multi-functional surfaces as well providing insights into the bactericidal mechanism underlying inherently antimicrobial materials, while suggesting that nanotopology is related to the evolutionary development of a species through the demands of its behavioural repertoire. The underlying relationship between the processes of wetting, adhesion and capillarity of the lipid nanopillars and bactericidal efficiency suggests new prospects for purely mechano-responsive antibacterial surfaces.

This work reports on the synthesis of titanium bone implants functionalized with nanoparticles (NPs) containing Arg-Gly-Asp-Cys peptide (RGDC) and shows the adhesion behavior of cells seeded on these materials. RGDC peptides were first conjugated to a norbornenyl-poly(ethylene oxide) macromonomer (Nb-PEO). Then, functional NPs with a size of ∼300 nm and constituted of polynorbornene core surrounded by poly(ethylene oxide) shell were prepared by ring-opening metathesis polymerization in dispersed medium. The grafting density of these NPs on the titanium surface is up to 2 NPs·μm(-2) (80 pmol of RGDC per cm(-2) of NP surface). Cell adhesion was evaluated using preosteoblast cells (MC3T3-E1). Results of cells cultured for 24 h showed that materials grafted with NPs functionalized with RGDC peptides enhance specific cell adhesion and can create filopodia-like structures among NP sites by stressing the cells.

Despite many years of in vitro research confirming the effectiveness of RGD in promoting cell attachment to a wide variety of biomaterials, animal studies evaluating tissue responses to implanted RGD-functionalized substrates have yielded more variable results. The goals of this report are to present some of the reasons why cell culture studies may not always reliably predict in vivo responses, and more importantly, to highlight potential applications that may benefit from the use of RGD peptides.Copyright © 2011 Elsevier Ltd. All rights reserved.

Medical device-associated infections account for a large proportion of hospital-acquired infections. A variety of opportunistic pathogens can cause implant infections, depending on the type of the implant and on the anatomical site of implantation. The success of these versatile pathogens depends on rapid adhesion to virtually all biomaterial surfaces and survival in the hostile host environment. Biofilm formation on implant surfaces shelters the bacteria and encourages persistence of infection. Furthermore, implant-infecting bacteria can elude innate and adaptive host defences as well as biocides and antibiotic chemotherapies. In this Review, we explore the fundamental pathogenic mechanisms underlying implant infections, highlighting orthopaedic implants and Staphylococcus aureus as a prime example, and discuss innovative targets for preventive and therapeutic strategies.

Quorum sensing is the regulation of gene expression in response to fluctuations in cell-population density. Quorum sensing bacteria produce and release chemical signal molecules called autoinducers that increase in concentration as a function of cell density. The detection of a minimal threshold stimulatory concentration of an autoinducer leads to an alteration in gene expression. Gram-positive and Gram-negative bacteria use quorum sensing communication circuits to regulate a diverse array of physiological activities. These processes include symbiosis, virulence, competence, conjugation, antibiotic production, motility, sporulation, and biofilm formation. In general, Gram-negative bacteria use acylated homoserine lactones as autoinducers, and Gram-positive bacteria use processed oligo-peptides to communicate. Recent advances in the field indicate that cell-cell communication via autoinducers occurs both within and between bacterial species. Furthermore, there is mounting data suggesting that bacterial autoinducers elicit specific responses from host organisms. Although the nature of the chemical signals, the signal relay mechanisms, and the target genes controlled by bacterial quorum sensing systems differ, in every case the ability to communicate with one another allows bacteria to coordinate the gene expression, and therefore the behavior, of the entire community. Presumably, this process bestows upon bacteria some of the qualities of higher organisms. The evolution of quorum sensing systems in bacteria could, therefore, have been one of the early steps in the development of multicellularity.

This manuscript describes the fabrication of arrays of spatially confined chambers embossed in a layer of poly(ethylene glycol) diacrylate (PEGDA) and their application to studying quorum sensing between communities of Pseudomonas aeruginosa. We hypothesized that biofilms may produce stable chemical signaling gradients in close proximity to surfaces, which influence the growth and development of nearby microcolonies into biofilms. To test this hypothesis, we embossed a layer of PEGDA with 1.5-mm wide chambers in which P. aeruginosa biofilms grew, secreted homoserine lactones (HSLs, small molecule regulators of quorum sensing), and formed spatial and temporal gradients of these compounds. In static growth conditions (i.e., no flow), nascent biofilms secreted N-(3-oxododecanoyl) HSL that formed a gradient in the hydrogel and was detected by P. aeruginosa cells that were ≤8 mm away. Diffusing HSLs increased the growth rate of cells in communities that were <3 mm away from the biofilm, where the concentration of HSL was >1 μM, and had little effect on communities farther away. The HSL gradient had no observable influence on biofilm structure. Surprisingly, 0.1-10 μM of N-(3-oxododecanoyl) HSL had no effect on cell growth in liquid culture. The results suggest that the secretion of HSLs from a biofilm enhances the growth of neighboring cells in contact with surfaces into communities and may influence their composition, organization, and diversity.

The modulation of bacterial communication to potentiate the effect of existing antimicrobial drugs is a promising alternative to the development of novel antibiotics. In the present study, we synthesized 58 analogues of hamamelitannin (HAM), a quorum sensing inhibitor and antimicrobial potentiator. These efforts resulted in the identification of an analogue that increases the susceptibility of Staphylococcus aureus towards antibiotics in vitro, in Caenorhabditis elegans, and in a mouse mammary gland infection model, without showing cytotoxicity.© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The saeRS two-component regulatory system regulates transcription of multiple virulence factors in Staphylococcus aureus. In the present study, we demonstrated that the saePQRS region in Staphylococcus epidermidis is transcriptionally regulated in a temporal manner and is arranged in a manner similar to that previously described for S. aureus. Studies using a mouse foreign body infection model demonstrated that the virulence of strain 1457 and the virulence of a mutant, strain 1457 saeR, were statistically equivalent. However, histological analyses suggested that the polymorphonuclear neutrophil response at 2 days postinfection was significantly greater in 1457-infected mice than in 1457 saeR-infected mice, demonstrating that SaeR influences the early, acute phases of infection. Microarray analysis demonstrated that a saeR mutation affected the transcription of 65 genes (37 genes were upregulated and 28 genes were downregulated); in particular, 8 genes that facilitate growth under anaerobic conditions were downregulated in 1457 saeR. Analysis of growth under anaerobic conditions demonstrated that 1457 saeR had a decreased growth rate compared to 1457. Further metabolic experiments demonstrated that 1457 saeR had a reduced capacity to utilize nitrate as a terminal electron acceptor and exhibited increased production of lactic acid in comparison to 1457. These data suggest that in S. epidermidis SaeR functions to regulate the transition between aerobic growth and anaerobic growth. In addition, when grown anaerobically, 1457 saeR appeared to compensate for the redox imbalance created by the lack of electron transport-mediated oxidation of NADH to NAD+ by increasing lactate dehydrogenase activity and the subsequent oxidation of NADH.

Widespread antibiotic resistance among important bacterial pathogens such as Staphylococcus aureus calls for alternative routes of drug development. Interfering with crucial virulence determinants is considered a promising new approach to control bacterial infection. Phenol-soluble modulins (PSMs) are peptide toxins with multiple key roles in pathogenesis and have a major impact on the ability of highly virulent S. aureus to cause disease. However, targeting PSMs for therapeutic intervention is hampered by their multitude and diversity. Here we report that an ATP-binding cassette transporter with previously unknown function is responsible for the export of all PSMs, thus representing a single target for complete obstruction of PSM production. The transporter had a strong effect on virulence phenotypes, such as neutrophil lysis, and the extent of its effect on the development of S. aureus infection was similar to that of the sum of all PSMs. Notably, the transporter was essential for bacterial growth. Furthermore, it contributed to producer immunity toward secreted PSMs and defense against PSM-mediated bacterial interference. Our study reveals a noncanonical, dedicated secretion mechanism for an important class of toxins and identifies this mechanism as a comprehensive potential target for the development of drugs to efficiently inhibit the growth and virulence of pathogenic staphylococci.

Polycationic nanoparticles show biocompatible, broad-spectrum bactericidal properties in vitro and in vivo when incorporated in denture lining material post-maxillectomy in head and neck cancer patients. In the present study, the synthesized Crosslinked quaternary ammonium polyethylenimine nanoparticles were found to have a strong bactericidal activity against a wide variety of microorganisms rapidly killing bacterial cells when incorporated at small concentrations into soft lining materials without compromising mechanical and biocompatibility properties. This appears advantageous over conventional released antimicrobials with regard to in vivo efficacy and safety, and may provide a convenient platform for the development of non-released antimicrobials. This is a crucial issue when it comes to giving an answer to the serious and life-threatening problems of contaminations in immunocompromised patients such as orofacial cancer patient.Copyright © 2014 Elsevier Ltd. All rights reserved.

Antimicrobial peptides are an abundant and diverse group of molecules that are produced by many tissues and cell types in a variety of invertebrate, plant and animal species. Their amino acid composition, amphipathicity, cationic charge and size allow them to attach to and insert into membrane bilayers to form pores by 'barrel-stave', 'carpet' or 'toroidal-pore' mechanisms. Although these models are helpful for defining mechanisms of antimicrobial peptide activity, their relevance to how peptides damage and kill microorganisms still need to be clarified. Recently, there has been speculation that transmembrane pore formation is not the only mechanism of microbial killing. In fact several observations suggest that translocated peptides can alter cytoplasmic membrane septum formation, inhibit cell-wall synthesis, inhibit nucleic-acid synthesis, inhibit protein synthesis or inhibit enzymatic activity. In this review the different models of antimicrobial-peptide-induced pore formation and cell killing are presented.

Anti-infective biomaterials need to be tailored according to the specific clinical application. All their properties have to be tuned to achieve the best anti-infective performance together with safe biocompatibility and appropriate tissue interactions. Innovative technologies are developing new biomaterials and surfaces endowed with anti-infective properties, relying either on antifouling, or bactericidal, or antibiofilm activities. This review aims at thoroughly surveying the numerous classes of antibacterial biomaterials and the underlying strategies behind them. Bacteria repelling and antiadhesive surfaces, materials with intrinsic antibacterial properties, antibacterial coatings, nanostructured materials, and molecules interfering with bacterial biofilm are considered. Among the new strategies, the use of phages or of antisense peptide nucleic acids are discussed, as well as the possibility to modulate the local immune response by active cytokines. Overall, there is a wealth of technical solutions to contrast the establishment of an implant infection. Many of them exhibit a great potential in preclinical models. The lack of well-structured prospective multicenter clinical trials hinders the achievement of conclusive data on the efficacy and comparative performance of anti-infective biomaterials. © 2013 Elsevier Ltd. All rights reserved.

Micro-porous TiO2 coatings co-doped with Zn2+ and Ag nanoparticles were fabricated on Ti by micro-arc oxidation (MAO) for 0.5, 1.5, 2 and 4?min, respectively. The evolutions of morphology and phase component of the coating as a function of processing time were investigated. The microstructure of the 2?min treated coating was further observed by transmission electron microscopy to explore the coating formation mechanism. The amounts of Ag and Zn released from the 2?min treated coating were measured and the antibacterial properties of the coatings against Staphylococcus aureus (S.?aureus) were also investigated. The obtained results showed that with prolonged MAO time, the contents of Ag and Zn on the coating surfaces increased. All the coatings were micro-porous with pore diameters of 1-4?μm; however, some pores were blocked by deposits on the 4?min treated coating. The 2?min treated coating was composed of amorphous TiO2, anatase, rutile, ZnO, Zn2TiO4 and homogenously distributed Ag nanoparticles. After immersion, Zn2+, Ag+, Ti2+ and Ca2+ were released from the coating and with the immersion time prolonged, the accumulated concentrations of these ions increased. After immersion for 36 weeks, the accumulated Zn2+ and Ag+ concentrations were 6.88 and 0.684?ppm, respectively, which are higher than the minimal inhibitory concentration but much lower than the cytotoxic concentration. Compared with polished Ti control, the coatings co-doped with Zn2+ and Ag nanoparticles significantly inhibited the adhesions of S.?aureus and reduced the amounts of planktonic bacteria in culture medium, indicating that the Zn and Ag co-doped TiO2 could be a bio-adaptable coating for long-lasting anti-microbial performance.

The current research is devoted to the study of the modification of the titanium implants by the micro-arc oxidation with bioactive calcium phosphate coatings containing Ag or Sr and Si elements. The coatings' microstructure, phase composition, morphology, physicochemical and biological properties were examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDX) and X-ray diffraction (XRD). Ag-containing and Sr-Si-incorporated coatings were formed in alkaline and acid electrolytes, respectively. The formation of the coatings occurred at different ranges of the applied voltages, which led to the significant difference in the coatings properties. The trace elements Ag, Sr and Si participated intensively in the plasma-chemical reactions of the micro-arc coatings formation. Ag-containing coatings demonstrated strong antibacterial effect against AТСС 6538-P. MTT test with 3T3-L1 fibroblasts showed no cytotoxicity appearance on Sr-Si-incorporated coatings.

Silver is widely used as a biocidal agent in ointments and wound dressings. However, it has also been associated with tissue toxicity and impaired healing. In vitro characterization has also revealed that typical loadings of silver employed in ointments and dressings (approximately 100 microg/cm(2)) lead to cytotoxicity. In this paper, we report the results of an initial study that sought to determine if localization of carefully controlled loadings of silver nanoparticles within molecularly thin films immobilized on surfaces can lead to antimicrobial activity without inducing cytotoxicity. Polymeric thin films of poly(allylamine hydrochloride) (PAH) and poly(acrylic acid) (PAA) were prepared by layer-by-layer deposition and loaded with approximately 0.4 microg/cm(2) to approximately 23.6 microg/cm(2) of silver nanoparticles. Bacterial killing efficiencies of the silver-loaded films were investigated against Staphylococcus epidermidis, a gram-positive bacterium, and it was determined that as little as approximately 0.4 microg/cm(2) of silver in the polymeric films caused a reduction of 6log(10)CFU/mL (99.9999%) bacteria in suspensions incubated in contact with the films (water-borne assays). Significantly, whereas the antibacterial films containing high loadings of silver were found to be toxic to a murine fibroblast cell line (NIH-3T3), the polymeric films containing approximately 0.4 microg/cm(2) of silver were not toxic and allowed attachment, and growth of the mammalian cells. Thus, the results of this study go beyond prior reports by identifying silver-impregnated, polymeric thin films that are compatible with in vitro mammalian cell culture yet exhibit antibacterial activity. These results support the hypothesis that localization of carefully controlled loadings of silver nanoparticles within molecularly thin polymeric films can lead to antimicrobial activity without cytotoxicity. More broadly, this strategy of modifying surfaces with minimal loadings of bioactive molecules indicates the basis of approaches that may permit management of microbial burden in wound beds without impairment of wound healing.

Two frequent problems are associated with the titanium surfaces of bone/dental implants: lack of native tissue integration and associated infection. These problems have prompted a significant body of research regarding the modification of these surfaces. The present study describes a hydrothermal treatment for the fabrication of strontium (Sr) and silver (Ag) loaded nanotubular structures with different tube diameters on titanium surfaces. The Sr loading from a Sr(OH)2 solution was regulated by the size of the inner diameter of the titanium nanotubes (NT) (30nm or 80nm, formed at 10V or 40V, respectively). The quantity of Ag was adjusted by immersing the samples in 1.5 or 2.0M AgNO3 solutions. Sr and Ag were released in a controllable and prolonged matter from the NT-Ag.Sr samples, with negligible cytotoxicity. Prominent antibacterial activity was observed due to the release of Ag. Sr incorporation enhanced the initial cell adhesion, migration, and proliferation of preosteoblast MC3T3-E1 cells. Sr release also up-regulated the expression of osteogenic genes and induced mineralization, as suggested by the presence of more mineralized calcium nodules in cells cultured on NT-Ag.Sr surfaces. In vivo experiments showed that the Sr-loaded samples accelerated the formation of new bone in both osteoporosis and bone defect models, as confirmed by X-ray, Micro-CT evaluation, and histomorphometric analysis of rats implanted with NT-Ag.Sr samples. The antibacterial activity and outstanding osteogenic properties of NT-Ag.Sr samples highlight their excellent potential for use in clinical applications.Two frequent problems associated with Ti surfaces, widely used in orthopedic and dental arenas, are their lack of native tissue integration and risk of infection. We describe a novel approach for the fabrication of strontium (Sr) and silver (Ag) loaded nanotubular structures on titanium surfaces. A relevant aspect of this work is the demonstration of long-lasting and controllable Ag release, leading to excellent antibacterial and anti-adherent properties against methicillin-resistant Staphylococcus aureus (MRSA), and Gram-negative bacteria such as Escherichia coli. The extended release of Sr accelerates the filling of bone defects by improving the repair of damaged cortical bone and increasing trabecular bone microarchitecture. Our results highlight the potential of Sr and Ag loaded nanotubular structures for use in clinical applications.Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Antibiotic-resistant bacteria are frequently involved in implant-associated infections (IAIs), making the treatment of these infections even more challenging. Therefore, multifunctional implant surfaces that simultaneously possess antibacterial activity and induce osseointegration are highly desired in order to prevent IAIs. The incorporation of multiple inorganic antibacterial agents onto the implant surface may aid in generating synergistic antibacterial behavior against a wide microbial spectrum while reducing the occurrence of bacterial resistance. In this study, porous titanium implants synthesized by selective laser melting (SLM) were biofunctionalized with plasma electrolytic oxidation (PEO) using electrolytes based on Ca/P species as well as silver and zinc nanoparticles in ratios from 0 to 100% that were tightly embedded into the growing titanium oxide layer. After the surface bio-functionalization process, silver and zinc ions were released from the implant surfaces for at least 28 days resulting in antibacterial leaching activity against methicillin-resistant Staphylococcus aureus (MRSA). Furthermore, the biofunctionalized implants generated reactive oxygen species, thereby contributing to antibacterial contact-killing. While implant surfaces containing up to 75% silver and 25% zinc nanoparticles fully eradicated both adherent and planktonic bacteria in vitro as well as in an ex vivo experiment performed using murine femora, solely zinc-bearing surfaces did not. The minimum inhibitory and bactericidal concentrations determined for different combinations of both types of ions confirmed the presence of a strong synergistic antibacterial behavior, which could be exploited to reduce the amount of required silver ions by two orders of magnitude (i.e., 120 folds). At the same time, the zinc bearing surfaces enhanced the metabolic activity of pre-osteoblasts after 3, 7, and 11 days. Altogether, implant biofunctionalization by PEO with silver and zinc nanoparticles is a fruitful strategy for the synthesis of multifunctional surfaces on orthopedic implants and the prevention of IAIs caused by antibiotic-resistant bacteria. STATEMENT OF SIGNIFICANCE: Implant-associated infections are becoming increasingly challenging to treat due to growing antibiotic resistance against antibiotics. Here, we propose an alternative approach where silver and zinc nanoparticles are simultaneously used for the biofunctionalization of rationally designed additively manufactured porous titanium. This combination of porous design and tailored surface treatment allows us to reduce the amount of required silver nanoparticles by two orders of magnitude, fully eradicate antibiotic-resistant bacteria, and enhance the osteogenic behavior of pre-osteoblasts. We demonstrate that the resulting implants display antibacterial activity in vitro and ex vivo against methicillin-resistant Staphylococcus aureus.Copyright © 2020 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

As a mussel-inspired material, polydopamine (PDA), possesses many properties, such as a simple preparation process, good biocompatibility, strong adhesive property, easy functionalization, outstanding photothermal conversion efficiency, and strong quenching effect. PDA has attracted increasingly considerable attention because it provides a simple and versatile approach to functionalize material surfaces for obtaining a variety of multifunctional nanomaterials. In this review, recent significant research developments of PDA including its synthesis and polymerization mechanism, physicochemical properties, different nano/microstructures, and diverse applications are summarized and discussed. For the sections of its applications in surface modification and biomedicine, we mainly highlight the achievements in the past few years (2016-2019). The remaining challenges and future perspectives of PDA-based nanoplatforms are discussed rationally at the end. This timely and overall review should be desirable for a wide range of scientists and facilitate further development of surface coating methods and the production of PDA-based materials.