Diego Mantovani

Small-diameter vascular tissue engineering

Vascular occlusion remains the leading cause of death in Western countries, despite advances made in balloon angioplasty and conventional surgical intervention. Vascular surgery, such as CABG surgery, arteriovenous shunts, and the treatment of congenital anomalies of the coronary artery and pulmonary tracts, requires biologically responsive vascular substitutes. Autografts, particularly saphenous vein and internal mammary artery, are the gold-standard grafts used to treat vascular occlusions. Prosthetic grafts have been developed as alternatives to autografts, but their low patency owing to short-term and intermediate-term thrombosis still limits their clinical application. Advances in vascular tissue engineering technology—such as self-assembling cell sheets, as well as scaffold-guided and decellularized-matrix approaches—promise to produce responsive, living conduits with properties similar to those of native tissue. Over the past decade, vascular tissue engineering has become one of the fastest-growing areas of research, and is now showing some success in the clinic.

Antibacterial Coatings: Challenges, Perspectives, and Opportunities

Antibacterial coatings are rapidly emerging as a primary component of the global mitigation strategy of bacterial pathogens. Thanks to recent concurrent advances in materials science and biotechnology methodologies, and a growing understanding of environmental microbiology, an extensive variety of options are now available to design surfaces with antibacterial properties. However, progress towards a more widespread use in clinical settings crucially depends on addressing the key outstanding issues. We review release-based antibacterial coatings and focus on the challenges and opportunities presented by the latest generation of these materials. In particular, we highlight recent approaches aimed at controlling the release of antibacterial agents, imparting multi-functionality, and enhancing long-term stability.

Long-term clinical study and multiscale analysis of in vivo biodegradation mechanism of Mg alloy

Significance In the past decade, countless studies have been performed to control the mechanical and corrosion property of magnesium-based alloy, which degrades in the physiological environment, to overcome the flaws of the inert implant materials and shift the paradigm of conventional bone fixation devices. Controlled degradation of Mg-5wt%Ca-1wt%Zn alloy results in the formation of biomimicking calcification matrix at the degrading interface to initiate the bone formation process. This process facilitates early bone healing and allows the complete replacement of biodegradable Mg implant by the new bone within 1 y of implantation, as demonstrated in 53 cases of successful long-term clinical study. There has been a tremendous amount of research in the past decade to optimize the mechanical properties and degradation behavior of the biodegradable Mg alloy for orthopedic implant. Despite the feasibility of degrading implant, the lack of fundamental understanding about biocompatibility and underlying bone formation mechanism is currently limiting the use in clinical applications. Herein, we report the result of long-term clinical study and systematic investigation of bone formation mechanism of the biodegradable Mg-5wt%Ca-1wt%Zn alloy implant through simultaneous observation of changes in element composition and crystallinity within degrading interface at hierarchical levels. Controlled degradation of Mg-5wt%Ca-1wt%Zn alloy results in the formation of biomimicking calcification matrix at the degrading interface to initiate the bone formation process. This process facilitates early bone healing and allows the complete replacement of biodegradable Mg implant by the new bone within 1 y of implantation, as demonstrated in 53 cases of successful long-term clinical study.

Novel Zn-based alloys for biodegradable stent applications: Design, development and in vitro degradation

The search for a degradable metal simultaneously showing mechanical properties equal or higher to that of stainless steel and uniform degradation is still an open challenge. Several magnesium-based alloys have been studied, but their degradation rate has proved to be too fast and rarely homogeneous. Fe-based alloys show appropriate mechanical properties but very low degradation rate. In the present work, four novel Zn-Mg and two Zn-Al binary alloys were investigated as potential biodegradable materials for stent applications. The alloys were developed by casting process and homogenized at 350°C for 48h followed by hot extrusion at 250°C. Tube extrusion was performed at 300°C to produce tubes with outer/inner diameter of 4/1.5mm as precursors for biodegradable stents. Corrosion tests were performed using Hanks׳ modified solution. Extruded alloys exhibited slightly superior corrosion resistance and slower degradation rate than those of their cast counterparts, but all had corrosion rates roughly half that of a standard purity Mg control. Hot extrusion of Zn-Mg alloys shifted the corrosion regime from localized pitting to more uniform erosion, mainly due to the refinement of second phase particles. Zn-0.5Mg is the most promising material for stent applications with a good combination of strength, ductility, strain hardening exponent and an appropriate rate of loss of mechanical integrity during degradation. An EBSD analysis in the vicinity of the laser cut Zn-0.5Mg tube found no grain coarsening or texture modification confirming that, after laser cutting, the grain size and texture orientation of the final stent remains unchanged. This work shows the potential for Zn alloys to be considered for stent applications.

Polydopamine as an intermediate layer for silver and hydroxyapatite immobilisation on metallic biomaterials surface

Hydroxyapatite (HA) coated implant is more susceptible to bacterial infection as the micro-structure surface which is beneficial for osseointegration, could also become a reservoir for bacterial colonisation. The aim of this study was to introduce the antibacterial effect of silver (Ag) to the biomineralised HA by utilising a polydopamine film as an intermediate layer for Ag and HA immobilisation. Sufficient catechol groups in polydopamine were required to bind chemically stainless steel 316 L, Ag and HA elements. Different amounts of Ag nanoparticles were metallised on the polydopamine grafted stainless steel by varying the immersion time in silver nitrate solution from 12 to 24 h. Another polydopamine layer was then formed on the metallised film, followed by surface biomineralisation in 1.5 Simulated Body Fluid (SBF) solution for 3 days. Several characterisation techniques including X-Ray Photoelectron Spectroscopy, Atomic Force Microscopy, Scanning Electron Microscopy and Contact Angle showed that Ag nanoparticles and HA agglomerations were successfully immobilised on the polydopamine film through an element reduction process. The Ag metallisation at 24 h has killed the viable bacteria with 97.88% of bactericidal ratio. The Ag was ionised up to 7 days which is crucial to prevent bacterial infection during the first stage of implant restoration. The aged functionalised films were considered stable due to less alteration of its chemical composition, surface roughness and wettability properties. The ability of the functionalised film to coat complex and micro scale metal make it suitable for dental and orthopaedic implants application.

Process of prototyping coronary stents from biodegradable fe-mn alloys

Biodegradable stents are considered to be a recent innovation, and their feasibility and applicability have been proven in recent years. Research in this area has focused on materials development and biological studies, rather than on how to transform the developed biodegradable materials into the stent itself. Currently available stent technology, the laser cutting-based process, might be adapted to fabricate biodegradable stents. In this work, the fabrication, characterization and testing of biodegradable Fe-Mn stents are described. A standard process for fabricating and testing stainless steel 316L stents was referred to. The influence of process parameters on the physical, metallurgical and mechanical properties of the stents, and the quality of the produced stents, were investigated. It was found that some steps of the standard process such as laser cutting can be directly applied, but changes to parameters are needed for annealing, and alternatives are needed to replace electropolishing.

Fucoidan in a 3D scaffold interacts with vascular endothelial growth factor and promotes neovascularization in mice

The aim of this study was to functionalize 3D porous cross-linked scaffolds with natural non-animal sulfated polysaccharide fucoidans in order to allow a delivery of vascular endothelial growth factor (VEGF) and potentiate its angiogenic activity. Microporous (20xa0μm) and macroporous (200xa0μm) scaffolds were functionalized with low, medium, or high molecular weight fucoidans (named LMWF, MMWF, and HMWF, respectively). In vitro, addition of fucoidans promoted endothelial progenitor cells proliferation in both micro- and macroporous scaffolds. While control scaffolds without fucoidans loaded with VEGF165 (100xa0ng) showed a fast burst release in PBS during the first 24xa0h, MMWF significantly reduced the VEGF165 release (pu2009<u20090.001). Surface plasmon resonance experiments confirmed a direct interaction between MMWF and VEGF165, characterized by an affinity KD (Kd/Ka) of 1u2009×u200910−9xa0M. In a subcutaneous angiogenesis model in mice, fucoidan functionalized scaffolds showed a more intense vascularization response than control groups. Expression of isolectin-B4 and α-smooth muscle actin, as well as confinement of erythrocytes, demonstrated the neoformed blood vessels functionality. There was a significant difference in neovessel area and neovessel density between MMWF scaffolds or VEGF165 scaffolds and MMWF+VEGF165 scaffolds (pu2009<u20090.001 for all cases). Here, we demonstrate that fucoidan sequesters VEGF165 and delivers biological cues promoting angiogenesis. In conclusion, this study shows that hydrogels functionalized with fucoidan can direct the formation of mature vasculature through a local release of VEGF165 and can be a useful tool in ischemic tissues to guide therapeutic angiogenesis.

Influence of cross-rolling on the micro-texture and biodegradation of pure iron as biodegradable material for medical implants.

Iron-based biodegradable metals have been shown to present high potential in cardiac, vascular, orthopaedic and dental in adults, as well as paediatric, applications. These require suitable mechanical properties, adequate biocompatibility while guaranteeing a low toxicity of degradation products. For example, in cardiac applications, stents need to be made by homogeneous and isotropic materials in order to prevent sudden failures which would impair the deployment site. Besides, the presence of precipitates and pores, chemical inhomogeneity or other anisotropic microstructural defects may trigger stress concentration phenomena responsible for the early collapse of the device. Metal manufacturing processes play a fundamental role towards the final microstructure and mechanical properties of the materials. The present work assesses the effect of mode of rolling on the micro-texture evolution, mechanical properties and biodegradation behaviour of polycrystalline pure iron. Results indicated that cross-rolled samples recrystallized with lower rates than the straight-rolled ones due to a reduction in dislocation density content and an increase in intensity of {100} crystallographic plane which stores less energy of deformation responsible for primary recrystallization. The degradation resulted to be more uniform for cross-rolled samples, while the corrosion rates of cross-rolled and straight-rolled samples did not show relevant differences in simulated body solution. Finally, this work shows that an adequate compromise between biodegradation rate, strength and ductility could be achieved by modulating the deformation mode during cold rolling.

Blood protein adsorption on sulfonated chitosan and κ-carrageenan films.

Many strategies have been reported to improve compatibility of biopolymers using chemical and physical modifications. One possibility is the introduction of sulfonate groups (R-SO3(-)) in the chitosan chain. Another biopolymer with similar characteristics to those of heparin is κ-carrageenan. This study proposed to investigate the application of these two polymers, based on their potential for globular protein adsorption (BSA and fibrinogen). Polymeric films of chitosan and κ-carrageenan were prepared; all films were characterized by elemental analyses, FTIR, XPS and SEM. Characterization techniques showed that the chitosan chain was modified and confirmed the existence of sulfonate groups, as well as in the κ-carrageenan chain, indicating surfaces with similar chemical properties to those of heparin. The effect of charge density was observed for each adsorption condition (BSA at pH 5.0 and 7.4). A more pronounced adsorption rate was observed at pH 5.0 than at pH 7.4 and equilibrium adsorption was achieved, in both cases, after approximately 20 min. The equilibrium data indicate a lower adsorption rate for the sulfonated chitosan film, in comparison to the other films. These results confirm the potential of modified chitosan for use in applications in which globular protein adsorption should be avoided.

Effect of grain sizes on mechanical properties and biodegradation behavior of pure iron for cardiovascular stent application

ABSTRACT Pure iron has been demonstrated as a potential candidate for biodegradable metal stents due to its appropriate biocompatibility, suitable mechanical properties and uniform biodegradation behavior. The competing parameters that control the safety and the performance of BMS include proper strength-ductility combination, biocompatibility along with matching rate of corrosion with healing rate of arteries. Being a micrometre-scale biomedical device, the mentioned variables have been found to be governed by the average grain size of the bulk material. Thermo-mechanical processing techniques of the cold rolling and annealing were used to grain-refine the pure iron. Pure Fe samples were unidirectionally cold rolled and then isochronally annealed at different temperatures with the intention of inducing different ranges of grain size. The effect of thermo-mechanical treatment on mechanical properties and corrosion rates of the samples were investigated, correspondingly. Mechanical properties of pure Fe samples improved significantly with decrease in grain size while the corrosion rate decreased marginally with decrease in the average grain sizes. These findings could lead to the optimization of the properties to attain an adequate biodegradation-strength-ductility balance.