Most common reasons for revision surgery of implants are loosening (65%), dislocation (9%) and infection (7%). The revision rate of hip replacement surgery is between 10-20%. Rates of implant loss for dental implants is 15% over a 5-year period. Biofilm formation is the major pathogenic factor. The project, driven by 7 dedicated complementary Hi-Tech SMEs, aims at gaining the scientific and processing knowledge to assist the SMEs in the development of the next generation multi-functional coatings for orthopaedic and dental implants and fixation devices for adequate implant fixation, bioresorbability, bioactivity, biocompatibility as well as biofilm formation inhibiting functionality, that should drastically reduce the currently experienced need for implant revisions due to loosening and infections. The envisaged radical innovations and major breakthroughs are: (a) development of new substrates and coatings with enhanced biocompatibility, (b) development of radically new or improvement of existing coating techniques for processing bioresorbable and biocompatible coatings with a graded interface and tailored porosity (c), in-depth understanding of implant substrate/coating/bone interfacial structure, design and engineering of optimal implant fixation, (d) knowledge on biofilm formation and inhibition (e) formulation and evaluation of biofilm inhibitors incorporated into the coating. The project addresses the integration of nanotechnologies, materials science and advanced technologies to improve health and quality of life of European citizens and creating wealth through novel knowledge-based and sustainable products (biomaterials) and processes (coatings), by means of fostering breakthrough applications through the integration of multi-disciplinary research developments. The project will contribute to a dynamic and competitive knowledge-based economy, sustainable development, and serves the needs of an SME-intensive sector.

Methicillin-resistant Staphylococcus aureus (MRSA), a virulent organism resistant to many drugs, is responsible for most nosocomial and community-acquired infections. It can cause life-threatening disease, and treatment options are limited. Effective and rapid diagnostics is a strategic key element in the campaign against the spread of MRSA, allowing infection surveillance and control measures as well as more efficient patient treatment, and overall decreased health cost. The MagRSA project aims at the development of a new diagnostics platform that will provide a fast, simple and accurate identification of MRSA from clinical samples. The diagnostic protocol relies on a new and clinically validated procedure that consists in a direct one-step enrichment of MRSA present in either nasal, inguinal or wound swabs, followed by DNA extraction of immunocaptured bacteria and their identification by multiplex sequences amplification using quantitative PCR. This protocol will be implemented with a simple 'hand-off¿ system based on: (1) advanced microfluidic magnetic nanoparticles manipulation technology allowing efficient capture and extraction of target bacteria and nucleic acids, and (2) novel strategies for the integration of full operations required for the entire nucleic acid analysis chain in a microfluidic platform. Steps as sample preparation signal amplification by multiplex PCR and detection of multiple genes simultaneously will be integrated and automated in single fluidic chip. MagRSA project will bring together high-tech SMEs and advanced hospital/academic laboratories that are mutually independent leaders in their fields with complementary competencies. The ultimate objective is to reach a pre-clinical validation of the proposed diagnostic tool that not only addresses the unmet need in MRSA diagnostics but also opens new potential for a broad range of applications in the emerging molecular diagnostics market.

The objective of FLUOROMAG will be to (a) produce noble metal nanoclusters or nanodots, (NDs), and core-shell (CSs) nanoparticles by a new method using controlled electrochemical techniques that ensure very uniform size distributions and transfer of this technology to a dedicated SME who will scale up the synthesis of these nanoparticles for commercial production as well as supply the consortium with NPs for characterization of their extinction, fluorescent and magnetic properties and the further development of diagnostic tests. (b) devise conjugation strategies to couple biomolecules to noble metal NDs and commercially available quantum dots, (QDs) to produce probes that can specifically target macromolecules such as proteins and DNA/RNA in vitro and in cells and tissues. We will take advantage of ND electrochemical synthesis to introduce specific molecules in the shells that permit efficient derivatization and coupling to biomolecules. (c) develop multiparametric diagnostic assays using combinations of bioconjugated QDs and noble metal NDs as novel, fluorescent probes, and bioconjugated noble metal nanoparticles as extinction probes. The goal is to achieve high sensitivity (down to single virus detection) in molecular and cellular recognition. New Hepatitis C, Dengue Fever and breast tumor assays are proposed that will monitor several antigens in multiplexed kinetic and end-point determinations. (d) develop a commercial, low-cost programmable array microscope (PAM) module for wide field microscopes with SME partner which utilizes a spatial light modulator to achieve high-speed sectioning and simultaneous measurement of multiple fluorescence modalities as a detection system for single and multiplexed diagnostic assays using nanoparticles developed in a-c for the health-care market.

WHO reports that tuberculosis results to millions of deaths or disabilities each year, especially in poorer areas of the planet. The problem is exacerbated by the AIDS epidemic that increases disease incidence in developed countries too. However in addition to tuberculosis, exposure to mycobacteria has also been linked to the pathogenesis of sarcoidosis and Crohn's disease that affect millions of people in Europe only. Diagnostic investigation of mycobacterial infections is hampered by the difficulty to detect in a specific manner low populations of mycobacteria or the immunology markers associated with the infections they cause. The NANOMYC project aims to develop a highly sensitive and specific, quantifiable detection system for molecular and immunology diagnostic markers associated with infection caused by M. tuberculosis complex (human and animal tuberculosis, implicated in sarcoidosis) and M. paratuberculosis (animal paratuberculosis, implicated in Crohn's disease). To this goal the consortium will combine nanotechnology and molecular biology incorporating the recent advances on the sequencing of mycobacterial genomes to routine diagnostics. Experts on mycobacteria genetics and nano-technology will produce a number of biomolecules that will be conjugated to a set of functionalized quantum dots with different oproelectrochemical characteristics. The NANOMYC assay will be developed in order to be applied for: a) in-field diagnostics using portable devices for evaluation of liquid samples, and b) manual and automated evaluation of solid samples using fluorescence resonance energy transfer. The final product will be a world innovation and it will target a market of several billion euro/year. Marketing of the NANOMYC assay will be used to sustain the project's administrative and financial platform and continue research activities in order to further the proposed technology for in-vivo diagnostic and therapeutic applications.

We will develop a new platform and assay for the detection of HIV p24 antigen in serum or blood. The advantages of a p24 test are that it can detect HIV in an early stage of the infection, before antibodies develop, and that it is quantitative. The DETECTHIV project aims at developing an extremely simple viral load test with only one reactant (grafted colloids). In a first phase, a magnetic nanoparticles assay will be developed for use in a microtiter plate with as goal a detection limit of 1 ng/ml. In a second phase, the use of nanoparticles on a microfluidic chip will permit to detect p24 to levels as low as 0.1 picog/ml, one to two orders of magnitude more sensitive than classical Enzyme Linked Immuno-Sorbent Assay (ELISA) p24 tests. Validation of our platform will be both done with recombinant p24 samples and patient samples. The principle of our test consists in optically detecting the formation of a colloidal gel of magnetic nanoparticles (`agglutination test'). The gel forms in a magnetic field under the presence of antigens that are capable of linking irreversibly two colloidal particles. Therefore the latter are grafted with antibodies that are specific to the p24 antigen. An agglutination test requires only one step with only one reactant and the detection is achieved through simple optical absorbance measurements, owing to the strong optical scattering modification when passing from nanometric colloids to the gelled state. In the microfluidic chip test, a sample solution of serum or blood is transported through a suspension of magnetic nanoparticles that are magnetically suspended within a microfluidic channel. When brought into a magnetic field, the particles will be able to approach each other, form chains and will be irreversibly linked if the p24 antigen is present. Subsequently, on-chip light scattering techniques will be used to quantify the concentration of permanent chains or clustered beads, which is proportional to the antigen concentration.'

The MuNanoVac STREP project will assess a new vaccine strategy to prevent HIV-1 infection based on a primo-vaccination using a biodegradable synthetic colloidal carrier made of poly-lactic acid (PLA) nanoparticles covered with adsorbed antigens. The aim is to demonstrate that PLA nanoparticles are a perfect mucosal vaccine vehicle, immunogenic for both arms of immunity, adaptable to many types of antigens, easy and simple to produce. Such nanoparticle-based vaccine carriers will allow targeting dentritic cells or transporting the vaccine through skin or mucosal epithelial barriers. To amplify the mucosal immune response, the project will investigate the potential use of immuno-modulator molecules associated with different immunization routes and schedules. Six main technical activities will be conducted in the project: 1) Elaborating a standardized formulation process of PLA nanoparticles with selected HIV-1 antigens, gag and trimeric gp140 2) Identifying the best adjuvant among four candidates to potentiate the mucosal immune responses induced by PLA formulations 3) Optimizing the uptake of nanoparticles by mucosa or skin and the orientation of the immune response 4) Defining the best immunization routes and schedules of nanoparticle formulation in presence or absence of immuno-modulators for immune responses 5) Testing safety and mucosal immunity of the two best combinations in rabbits and 6) Evaluating the vaccine strategy efficacy studies with rectal and vaginal challenge routes in non human primates. Additional activities will include dissemination and exploitation of results, IPR and societal issues. The MuNanoVac project will contribute to advancing a promising vaccine approach for HIV, that could prove versatile enough for application to other poverty-related diseases e.g. Tuberculosis. With the proposed vaccine candidate, the project gives Europe a tremendous opportunity to gain leadership in the use of biodegradable nanoparticles for vaccine carriers.

Outstanding progress has been made in recent years in developing novel structures and applications for direct fabrication of 3D nanosurfaces. However, exploitation is limited by lack of suitable manufacturing technologies. In this project we will develop innovative in-line high throughput technologies based on atmospheric pressure surface and plasma technologies. The two identified approaches to direct 3D nanostructuring are etching for manufacturing of nanostructures tailored for specific applications, and coating. Major impact areas were selected, demonstrating different application fields. Impact Area 1 focuses on structures for solar cell surfaces. Nanostructured surfaces have the potential to improve efficiencies of cells by up to 25% (rel), having dramatic impact on commercial viability. Impact Area 2 focuses on biocidal surface structures. Increasing concerns about infections leading to the conclusion, that only multi-action approaches for control of infection transfer can be effective. We plan to combine such surfaces with 3D nanostructures, which will both immobilise and deactivate pathogenic organisms on surfaces. Impact Area 3 is the direct growth of aligned carbon nanotubes on electrode surfaces. The material is under investigation for use in high load capacitors which are seen as key components for energy storage systems, e.g. for Hybrid Electric Vehicle. Impact Area 4 focuses on tailored interfaces to achieve durable adhesion on polymer surfaces by 3D nanostructuring and coating. Target is to reduce energy consumption by introducing lightweight materials. The N2P partners have been chosen to ensure a strong capability to exploit and disseminate the outcomes. Involved end-user industries represent high market value segments: photovoltaics, aeronautics, automotive, steel. The consortium includes 7 technology leading SMEs and 4 multi-national industries, cooperating with 9 institutes for industrial research and a public body from 8 European countries.

Congenital and acquired diseases of the heart are the leading causes of morbidity and mortality in the world today; 7.2 million people die each year due to coronary heart disease, being the first cause of mortality in population above 60 years old, and the second cause after HIV in world wide young population. There is an urgent demand for new methods to repair and replace damaged cardiovascular tissues. One of the most promising ways to achieve this goal is the development of regenerative therapies aided with novel intelligent nanobiomaterials such as bioactive scaffolds. The overall objective of this project is the development of innovative bioactive polymeric scaffolds able to guide tissue formation from dissociated stem cells, for engineering autologous cardiovascular replacements, namely vascular tissues, heart valves and cardiac muscle. Two different strategies will be followed to approach creating new engineered tissue: 1.In vitro tissue engineering: according to the most frequent tissue engineering paradigm, cells will be seeded on a scaffold composed of synthetic polymer or natural material and the tissue will be matured in vitro in a bioreactor, in order to obtain a construct that can be implanted in the appropriate anatomic location as a prosthesis; 2.In vivo tissue engineering: unseeded scaffolds that attract endogenous cells and control cell proliferation and differentiation will be implanted to repopulate and remodel an altered cardiovascular tissue. The strong innovative content of the project is in the realisation of multifunctional scaffolds which can guide complex cellular processes such as adhesion, proliferation and differentiation, processes fundamental for tissue regeneration. It is therefore necessary to design integrated material scaffolds and culture environments, which can appropriately confer biochemical, morphological, electrical and mechanical stimuli to a developing tissue.

The DIVINOCELL project will identify novel Gram-negative targets by exploiting the components of the divisome, their activities and interactions. It will also design selective assays for screening and will obtain a new class of antimicrobials: compounds to block bacterial division. New medicines to attack Gram-negative pathogens will decrease the burden of infectious disease and have a highly beneficial social and economic impact in Europe and beyond. Cell division is an essential and still underexploited process with excellent properties to yield new inhibitors to attack infection by blocking the proliferation of pathogens. Inhibitors directed against bacterial division targets, that are not present in eukaryotic cells, will be both effective and innocuous to humans and animals. In addition, as many of their structures will be based on interaction domains and synthetic scaffolds, they will generate resistance at levels lower than the present antibiotics. DIVINOCELL will apply existing and new knowledge on the molecular biology of Gram-negative cell division as well as novel analytical (nanodiscs), bioinformatic (molecular dynamics), structural (membrane protein crystals) and imaging (lanthanide staining) tools to exploit in the test tube the structures and interactions of targets in the divisome and the septum. DIVINOCELL will develop potent systematic screening assays and will use them to select compounds specifically tailored to inhibit the division of Gram-negatives (not precluding broad spectrum ones). Secondary activity and cell assays, based on the properties of bacterial division, will be generated to validate hits and advance them to leads. The medicinal properties of selected leads will be improved. The translational steps of the project will be developed by 4 SMEs in close collaboration with the 8 academic partners having well-proven expertise in molecular microbiology, protein chemistry, structural biology, biophysics, imaging and bioinformatics.

We have recently developed a bionanotechnology approach to vaccination (Reddy et al., Nature Biotechnology, 25, 1159-1164, 2007): degradable polymeric nanoparticles are designed that: (i) are so small that they can enter the lymphatic circulation by biophysical means; (ii) are efficiently taken up by a large fraction of dendritic cells (DCs) that are resident in the lymph node that drains the injection site; (iii) activate the complement cascade and provide a potent, yet safe, activation signal to those DCs; and (iv) thereby induce a potent, Th1 adaptive immune response to antigen bound to the nanoparticles, with the generation of both antibodies and cytotoxic T lymphocytes. In the present project, we focus on next-generation bionanotechnology vaccine platforms for vaccination. We propose three technological advances, and we propose to demonstrate those three advances in definitive models in the mouse. Specifically, we propose to (Specific Aim 1) evaluate the current approach of complement-mediated DC activation in breaking tolerance to a chronic viral infection (hepatitis B virus, HBV, targeting hepatitis B virus surface antigen, HBsAg) and to combine complement as a danger signal with other nanoparticle-borne danger signals to develop an effective bionanotechnological platform for therapeutic antiviral vaccination; (Specific Aim 2) to develop a new, ultrasmall nanoparticle implementation suitable for delivery of DNA to lymph node-resident DCs, also activating them, to enable more efficient DNA vaccination; and (Specific Aim 3) to develop an ultrasmall nanoparticle implementation suitable for delivery of DNA to DCs resident within the sublingual mucosa, also activating them, to enable efficient DNA mucosal vaccination. The Specific Aim addressing the oral mucosa will begin with HBsAg, to allow comparison to other routes of administration, and will then proceed to antigens from influenza A.