F. Javier Gil

Corrosion behaviour of commercially pure titanium shot blasted with different materials and sizes of shot particles for dental implant applications

It is well known that the osseointegration of the commercially pure titanium (c.p. Ti) dental implant is improved when the metal is shot blasted in order to increase its surface roughness. This roughness is colonised by bone, which improves implant fixation. However, shot blasting also changes the chemical composition of the implant surface because some shot particles remain adhered on the metal. The c.p. Ti surfaces shot blasted with different materials and sizes of shot particles were tested in order to determine their topographical features (surface roughness, real surface area and the percentage of surface covered by the adhered shot particles) and electrochemical behaviour (open circuit potential, electrochemical impedance spectroscopy and cyclic polarisation). The results demonstrate that the increased surface area of the material because of the increasing surface roughness is not the only cause for differences found in the electrochemical behaviour and corrosion resistance of the blasted c.p. Ti. Among other possible causes, those differences may be attributed to the compressive residual surface stresses induced by shot blasting. All the materials tested have an adequate corrosion and electrochemical behaviour in terms of its possible use as dental implant material.

Novel peptide-based platform for the dual presentation of biologically active peptide motifs on biomaterials

Biofunctionalization of metallic materials with cell adhesive molecules derived from the extracellular matrix is a feasible approach to improve cell-material interactions and enhance the biointegration of implant materials (e.g., osseointegration of bone implants). However, classical biomimetic strategies may prove insufficient to elicit complex and multiple biological signals required in the processes of tissue regeneration. Thus, newer strategies are focusing on installing multifunctionality on biomaterials. In this work, we introduce a novel peptide-based divalent platform with the capacity to simultaneously present distinct bioactive peptide motifs in a chemically controlled fashion. As a proof of concept, the integrin-binding sequences RGD and PHSRN were selected and introduced in the platform. The biofunctionalization of titanium with this platform showed a positive trend towards increased numbers of cell attachment, and statistically higher values of spreading and proliferation of osteoblast-like cells compared to control noncoated samples. Moreover, it displayed statistically comparable or improved cell responses compared to samples coated with the single peptides or with an equimolar mixture of the two motifs. Osteoblast-like cells produced higher levels of alkaline phosphatase on surfaces functionalized with the platform than on control titanium; however, these values were not statistically significant. This study demonstrates that these peptidic structures are versatile tools to convey multiple biofunctionality to biomaterials in a chemically defined manner.

A bioactive elastin-like recombinamer reduces unspecific protein adsorption and enhances cell response on titanium surfaces.

We present the immobilization on synthetic substrates of elastin-like recombinamers (ELR) that combine a bioactive motif for cell adhesion with protein antifouling properties. Physical adsorption of the recombinamers and covalent-grafting through organosilane chemistry were investigated. The biochemically-modified surfaces were thoroughly characterized and tested for protein absorption in serum by fluorescence-labelling, XPS, Ellipsometry, and OWLS. The ELR were successfully grafted and stable, even upon mechanical stresses; being the covalent bonding favourable over physical adsorption. The coated metal surfaces exhibited excellent reduction of serum protein adsorption (9 ng/cm(2)) compared to the bare metal surface (310 ng/cm(2)). Non-specific protein adsorption may mask the introduced bioactive motifs; therefore, the bioactivated surfaces should display serum-protein antifouling properties. Finally, improved hMSCs response was assessed on the bioactivated substrates. In summary, the coatings simultaneously displayed anti-fouling and bioactive properties. These studies investigated key factors to enhance tissue material interactions fundamental for the design of bioactive devices and future biomedical applications.

Biofunctionalization strategies on tantalum-based materials for osseointegrative applications

The use of tantalum as biomaterial for orthopedic applications is gaining considerable attention in the clinical practice because it presents an excellent chemical stability, body fluid resistance, biocompatibility, and it is more osteoconductive than titanium or cobalt-chromium alloys. Nonetheless, metallic biomaterials are commonly bioinert and may not provide fast and long-lasting interactions with surrounding tissues. The use of short cell adhesive peptides derived from the extracellular matrix has shown to improve cell adhesion and accelerate the implant’s biointegration in vivo. However, this strategy has been rarely applied to tantalum materials. In this work, we have studied two immobilization strategies (physical adsorption and covalent binding via silanization) to functionalize tantalum surfaces with a cell adhesive RGD peptide. Surfaces were used untreated or activated with either HNO3 or UV/ozone treatments. The process of biofunctionalization was characterized by means of physicochemical and biological methods. Physisorption of the RGD peptide on control and HNO3-treated tantalum surfaces significantly enhanced the attachment and spreading of osteoblast-like cells; however, no effect on cell adhesion was observed in ozone-treated samples. This effect was attributed to the inefficient binding of the peptide on these highly hydrophilic surfaces, as evidenced by contact angle measurements and X-ray photoelectron spectroscopy. In contrast, activation of tantalum with UV/ozone proved to be the most efficient method to support silanization and subsequent peptide attachment, displaying the highest values of cell adhesion. This study demonstrates that both physical adsorption and silanization are feasible methods to immobilize peptides onto tantalum-based materials, providing them with superior bioactivity.

Silver deposition on titanium surface by electrochemical anodizing process reduces bacterial adhesion of Streptococcus sanguinis and Lactobacillus salivarius

OBJECTIVES The aim of this study was to determine the antibacterial properties of silver-doped titanium surfaces prepared with a novel electrochemical anodizing process. MATERIAL AND METHODS Titanium samples were anodized with a pulsed process in a solution of silver nitrate and sodium thiosulphate at room temperature with stirring. Samples were processed with different electrolyte concentrations and treatment cycles to improve silver deposition. Physicochemical properties were determined by X-ray photoelectron spectroscopy, contact angle measurements, white-light interferometry, and scanning electron microscopy. Cellular cytotoxicity in human fibroblasts was studied with lactate dehydrogenase assays. The in vitro effect of treated surfaces on two oral bacteria strains (Streptococcus sanguinis and Lactobacillus salivarius) was studied with viable bacterial adhesion measurements and growth curve assays. Nonparametric statistical Kruskal-Wallis and Mann-Whitney U-tests were used for multiple and paired comparisons, respectively. Post hoc Spearmans correlation tests were calculated to check the dependence between bacteria adhesion and surface properties. RESULTS X-ray photoelectron spectroscopy results confirmed the presence of silver on treated samples and showed that treatments with higher silver nitrate concentration and more cycles increased the silver deposition on titanium surface. No negative effects in fibroblast cell viability were detected and a significant reduction on bacterial adhesion in vitro was achieved in silver-treated samples compared with control titanium. CONCLUSIONS Silver deposition on titanium with a novel electrochemical anodizing process produced surfaces with significant antibacterial properties in vitro without negative effects on cell viability.

Biofunctional polyethylene glycol coatings on titanium: an in vitro-based comparison of functionalization methods

Three methods for the production of Polyethylene glycol (PEG) coatings on titanium are compared, i.e. plasma polymerization, electrodeposition and silanization. The compared deposition methods presented similar wettability (hydrophilic coatings), chemical composition assessed by XPS and thickness around 1nm. The coatings lowered albumin adsorption and presented a decreased fibroblast, Streptococcus sanguinis and Lactobacillus salivarius adhesion. Immobilization of a cell adhesion peptide (RGD) presented a higher fibroblast adhesion and no alteration of the bacterial adhesion, giving three methods for the biofunctionalization of titanium for dental implants. The feasibility of each methodology is compared in terms of the process parameters in order to provide a guide for the election of the methodology.

Assessment and comparison of surface chemical composition and oxide layer modification upon two different activation methods on a cocrmo alloy

This study investigated the effect of two different activation methods on the surface chemical composition of a CoCrMo-alloy. The activation was performed with oxygen plasma (OP) or nitric acid (NA). The surface physical–chemical properties were thoroughly characterized by means of several analytical techniques: X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectrometry (ToF-SIMS), zinc-complex substitution technique, contact angle, and interferometry. The surface modification was evaluated by assessing contamination removal, the “active” hydroxyl groups (OH-act) present at the surface, the metal oxide ratio (CoyOx−/CryOx−) and changes in the chemical composition and topography of the oxide layer. XPS experimental data showed for both methods (OP and NA) a significant decrease of the carbon contents (C 1s) associated with contaminants and at the same time changes in the atomic composition of the oxide layer (O 1s). In addition, the O 1s XPS spectra showed differences between the percentage of OH− before and after OP or NA treatment, leading to the conclusion that both methods are effective for surface “cleaning” and activation. These results were further investigated and corroborated by ToF-SIMS analysis and zinc complex substitution technique. The general conclusion was that NA is more efficient in terms of contaminants removal and generation of accessible OH-act present at the surface and without altering the native metal oxide ratio (CoyOx−/CryOx−) considered to be essential for biocompatibility.

Regenerating bone via multifunctional coatings: the blending of cell integration and bacterial inhibition properties on the surface of biomaterials

In dentistry and orthopedics, it is well accepted that implant fixation is a major goal. However, an emerging concern is bacterial infection. Infection of metallic implants can be catastrophic and significantly reduce patient quality of life. Accordingly, in this work, we focus on multifunctional coatings to simultaneously address and mitigate both these problems. We have developed a tailor-made peptide-based chemical platform that integrates the well-known RGD cell adhesive sequence and the lactoferrin-derived LF1-11 antimicrobial peptide. The platform was covalently grafted on titanium via silanization and the functionalization process characterized by contact angle, XPS, and QCM-D. The presence of the platform statistically improved the adhesion, proliferation and mineralization of osteoblast-like cells compared to control surfaces. At the same time, colonization by representative bacterial strains was significantly reduced on the surfaces. Furthermore, the biological potency of the multifunctional platform was verified in a co-culture in vitro model. Our findings demonstrate that this multifunctional approach can be useful to functionalize biomaterials to both improve cell integration and reduce the risk of bacterial infection.

Mechanical properties of a new thermoplastic polymer orthodontic archwire.

A new thermoplastic polymer for orthodontic applications was obtained and extruded into wires with round and rectangular cross sections. We evaluated the potential of new aesthetic archwire: tensile, three point bending, friction and stress relaxation behaviour, and formability characteristics were assessed. Stresses delivered were generally slightly lower than typical beta-titanium and nickel-titanium archwires. The polymer wire has good instantaneous mechanical properties; tensile stress decayed about 2% over 2h depending on the initial stress relaxation for up to 120h. High formability allowed shape bending similar to that associated with stainless steel wires. The friction coefficients were lower than the metallic conventional archwires improving the slipping with the brackets. This new polymer could be a good candidate for aesthetic orthodontic archwires.

Surface immobilization and bioactivity of TGF-β1 inhibitor peptides for bone implant applications

TGF-β1 is the most related cytokine with the production of fibrotic tissue. It plays an important role on the production of collagen by fibroblasts and other types of cells. The inhibition of this cytokine reduces fibrosis in various types of tissue. Biofunctionalization of dental and orthopedic implants with biomolecules enables modification of the physical, chemical and biochemical properties of their surfaces to improve its biological and clinical performance. Our objective was to develop a reliable method to immobilize oligopeptides on Ti surfaces to obtain a surface with TGF-β1 inhibitory activity that will potentially minimize fibrotic encapsulation of implants during the process of osseointegration. We covalently immobilized TGF-β1 inhibitor P17-peptides on Ti surfaces and assessed by characterizing each step of the process that we successfully biofunctionalized the implant surfaces. High amounts of peptides were anchored and homogeneously distributed on the surfaces with mechanical and thermochemical stability after in vitro simulated challenges. Notably, the immobilized peptides retained their TGF-β1 inhibitory activity in vitro. Thus, these biofunctional coatings are potential candidates for inducing a fast and reliable osseointegration in vivo.