D2.2 Final report on Safety by Process Design implementation for the coating processes


The analysis of risk, detailed in the previous deliverable D2.1, allowed to identify the critical steps of the deposition process. In particular, the hot-spots, corresponding to the highest probability to release nanoparticles resulted: the ultrasonication step of sonochemical process and the atomization through ultrasound nozzles, during the spray-deposition process. Thus, we focused on these steps of the process and performed measurement campaigns on lab-scale plants and pilot lines, determining quantitatively the emission of particles, in order to estimate the exposure of workers, through inhalation
Many data on inhalation exposure expressed as particle number concentration, particle size distribution, lung deposited surface area (LDSA), were experimentally. Data processing, providing quantitative information for the risk estimation, allowed to compare, in some cases, the exposure level with human health guidance values, such as recommended exposure limit (REL) and to advance hypothesis in terms of design of a safer manufacturing process.

D3.4 Final report on the smart metabolic sensing for functional lifetime monitoring of the end products 


This final report corresponds to Task 3.5, which entailed the development of a smart bacterio-sensing textile with capacity to detect living bacteria on hospital fabrics. This task was split in three consecutives sub-tasks for simplification of the development process:

  • Sub-Task 3.5.1 Selection of the Sensing Probe: A number of electrochromic metabolic indicators with capacity to change of colour in presence of living bacteria were evaluated in solution. Prussian Blue was deeply studied and finally selected for presenting an intense change of colour (from deep blue to unicolor), low toxicity and being susceptible of implementation in textiles.
  • Sub-Task 3.5.2 Implementation in Textiles: Sonochemical coating was chosen initially, in agreement with the objectives of the project, providing coloured and quite homogenous coatings in both raw and antibacterial cotton-polyester samples (through previous coating of CuO-nanoparticles). Additionally, cyanotyping was evaluated for its simplicity, producing even more intensely-coloured, homogeneous and stable coatings.
  • Sub-Task 3.5.3 Evaluation of Smart Textile Function: Escherichia coli and Staphylococcus aureus were used as model gram-negative and gram-positive bacteria. In both cases, colour changes due to bacterial metabolism could be detected with the bare eye after 5-6 hours, while textiles incubated in buffer samples or samples with dead cells did not change of colour. Additionally, bacterial death resulted in the recovery of the initial colour of the textile through oxygen-mediated re-oxidation of Prussian Blue.

This smart bacterio-sensing technology, compatible with the antibacterial coatings proposed in PROTECT, was sensitive enough to indicate the end of the lifetime of bactericidal textiles, thus acting as an early warning system for prevention of nosocomial infections.

D3.5 Report and demonstration of the automation system for solution delivery and adjustment of pH, concentration and fabric speed of the three processes


The purpose of this document is to define the set of functional tests needed to validate the R2R and Batch machine. The tests
have been performed in the first step at Icone, and at end user manufacture KLOPMAN (R2R) and DEGANIA (BATCH).
All tests were done with water instead of the reagent solution. The functional tests are set on three parts: the components (sensors and motors) were tested individually; the subcomponents were tested individually and in the combined configuration. All tests were performed on the open and close loop configuration.

D3.6 Report on the up-scaled pre-commercial processes, including the monitoring and automation system processes


The implementation and full integration of spectroscopy-based technology for in situ monitoring of NPs concentration, size, and colloidal stability into the up-scaled pre-commercial processes is described. In order to increase the robustness and repeatability of the coating processes, spectrometers and pushbroom imaging (PBI) technology with spatial resolution of the spectral information are introduced into the three pre-commercial manufacturing lines with the aim of providing fundamental information via in situ measurements that serve as quality control and can equally be used for optimisation of the processes and end-products. The current deliverable lists the monitoring requirements of each pre-commercial manufacturing line, describes into detail the spectroscopy-based solution designed by INDA, shows the introduction and integration of the developed spectroscopic equipment in the process of the three plants and illustrates its functionality through data.

D4.2 Report on human nanotoxicity of the functional products according to the NanoSafety Cluster and OECD guidelines 


WP4, and in particular of the present deliverable, is to investigate the hazards deriving from the production and use of functional products achieved by the coating with different nanomaterials (NMs), like metal oxides (ZnO, CuO, Zn-doped CuO), TiO2/SiO2 nanocomposites and organic polymers (polypirrole, PPy).

After having identified the most representative exposure scenarios and the hazard determinants toward environmental and human health (D4.1), the deliverable 4.2 is mainly related to report on the results obtained from the nanotoxicity studies performed on the nanoparticles (NPs) and the coated nano-enabled products (NEPs), consisting in antimicrobial coated textiles, water depuration membranes and catheters, produced in the frameworks of PROTECT.  Considering that the main exposures to antimicrobials NPs have been predicted to be inhalation and skin contact, in the present document presents the results obtained from i) inhalation toxicity and exposure and ii) skin toxicity. Moreover, since potential NPs release may occur after leaching from antimicrobial-coated water depuration membranes, the impact on freshwater living organisms (i.e. zebrafish) has also been evaluated. The setup of the in-silico methodology to estimate the nano risk and the future suitable management strategies are also reported. The methods to evaluate the NPs potential toxicity have been set up, mainly following the OECD and NanoSafety Cluster (NSC) documents, reporting the test guidelines to investigate NPs toxicity after skin contact (OECD TG. 431 and 439), inhalation exposure and effects of NPs in workplaces (OECD guideline ENV/JM/MONO(2015); no standard in vitro test available for inhalation toxicology, NSC-suggested testing strategies) and in the aquatic environment respectively (OECD TG. 236). Since the NMs developed in the project are represented by almost completely new nanoformulations, especially when considering sonochemically synthesized MeOs, hybrid TiO2/SiO2 and PPy, and no data are available in literature for their toxicity, dedicated in vitro toxicity assays have been performed. The biological investigations involved in vitro tests on 2D monoculture of human lung cell lines (A549 in particular) and murine fibroblasts, as well as on 3D biological models, like human reconstructed epidermis and air-blood barrier (ABB). The (Fish Embryo acute Toxicity test (FET), using zebrafish, was also adopted.

By using the above described methods, it has been possible to define the hazard of antimicrobial NPs and NEPs toward humans and to generate useful data for the risk assessment frameworks

D5.2 Report on the anti-biofilm efficiency of the water treatment membranes used in water scarce area – Catania and Gela, in Sicily


Research on polymeric-based water treatment membranes was done at the beginning of the project, in order to choose the most suitable material and format for a further nanoparticle (NP)-based functional treatment. FN has chosen Polyamide membranes as chemically and physically stable, they do not hydrolyze in water and tolerate pH values included in a Range 3 – 11. They have excellent properties for water treatment and filtration but are very susceptible to particulate fouling and biofouling. These weaknesses provide a good opportunity for the NP-based sonochemical treatment to show its potential. First coating trials performed by BIU and further antimicrobial tests on treated membranes performed by CDB show that the NP-based coating treatment has an anti-proliferative characteristic of the bacterial colonies, which is evident at 37°C. The mechanical and chemical main issues that could negatively interact with the work of the treated membranes during their life cycle were assessed, too, by FN, KLO, CDB, UNIMIB and BIU. Main mechanical issues were related to potential breaking damage during the coating, and traction forces could damage the porosities making the filters ineffective. Main chemical issues were related to potential interactions between acid/basic media (used to maintain high filter performance) and the NP-based coating, that could be damaged or removed from the surface of the polymeric membrane. Treatment of the membranes with HCl did not negatively influence the antimicrobial and antiproliferative effect, but even in some cases shows a synergistic effect.

D7.4 Plan for using and disseminating the Know-how


The aim of this deliverable is to present the mid version of the dissemination and exploitation plan of the PROTECT project. It presents the actions that will be taken to disseminate the PROTECT project highlights. Different dissemination avenues are described in relation to the stakeholders to be reached out. Dissemination methods range from the Internet website and social media to journals and conferences. The project website is recognised as a key dissemination medium which brings together all other types of dissemination. Exploitation plan starts with brief explanation of legal boundaries. We propose a set of exploitation tools and identify potential customers of PROTECT solutions.

D2.1 Report on Safety by Process Design implementation for the coating processes


One of the objectives of PROTECT is to scale up the production of antibacterial textiles, obtained from the functionalization of fibers with bactericidal nanoparticles, in three pilots: 1) roll-to-roll continuous sonochemical coating; 2) roll-to-roll continuous spray coating; and 3) batch sonochemical functionalization. For each pilot, this deliverable 1) establishes criteria for estimating hazard and exposure for each nanoparticle and for each pilot; 2) defines bands (i.e.,categories) of risk that cover the full range of values given by hazard and exposure; and 3) proposes actions that minimize the risk specific to each band. By dissecting the production of antibacterial textile in each pilot, this deliverable identifies the crucial steps that must be considered in the design of future safe manufacturing. A conclusion common to all the production methods analyzed is that manual handling of chemicals and nanoparticles and open air processes lead to the highest risks for the health of workers: automation of these steps would reduce greatly the risks.

D7.9 Report on training courses for employees about the use of the control and monitoring systems


For a future intelligent manufacturing four toolboxes are needed to implement smart sensors into the production line of surface nanostructured antimicrobial and anti-biofilm textiles: toolbox 1: spectroscopy to obtain information on a molecular level,  toolbox 2: design of experiments to get independent information from each experiment, toolbox 3: multivariate data analysis to exploit the fundamental information of the huge data sets and toolbox 4: a newly developed pushbroom imaging system to get spatially resolved information about the morphology and chemistry of adhered nanoparticles. A concept has been developed to train personal in research institutions and staff in industry at the regular 6 months meetings as well as with special courses and practical onsite trainings during the development of the production line.

D7.16 Forth business plans for the new products (four business cases)


The fourth Business plan of the four products reports update data in relation to the social need the project aims to respond, market, analysis, state of the art of competing products, data on financial plan and products pricing policies. The Business plan realization is based also on a new data collection approach represented by direct market survey. This approach’s goal is twofold: on the one side it is to collect direct opinion and purchase attitude of the final costumers; on the other side id to introduce the first direct awareness action on the products development. In addition, Business Model Canvas for each of the four solution are included in the business plan with the aim of defining all aspects for each solution, analysing deeply and highlighting in details the key partners, potential customers, key resources, possible different value propositions for each sector, channels, costumer relations and revenue flows.

D2.3 Report on the three processes up-scaled for production of antimicrobial/anti-biofilm, and biocompatible products


This deliverable describes the progress towards the scale-up of the three processes developed in Protect: i) the continuous roll-to-roll (R2R) sonochemical coating, ii) the R2R spray coating using ultrasound nozzles, and iii) the batch sonochemical coating. Partner KLO carried out experiments on the existing R2R pilot to define and optimize the parameters required for a homogenous coating that will be further used in the new up-scaled machine. Partner CENTI optimized the spray R2R coating of cotton fabrics with photocatalytic and antimicrobial silica/titanium dioxide nanocomposites. The main parameters related with the spray deposition process were optimized following specific design-of-experiments trials, with two kinds of spray nozzles: pressurized and ultrasonic. Partners UPC and BIU optimized the batch coating processes for medical devices using metal oxide nanoparticles, combination of metaloxides and enzymes, or functional biomolecules.

D2.4 Report on the assembled pre-commercial coating lines at the industrial premises of the partners


This deliverable describes the progress in the construction and assembly of the three pilots developed in Protect: i) the continuous roll-to-roll (R2R) sonochemical machine, ii) the R2R spray coating line using ultrasound nozzles, and iii) the batch sonochemical machine. The building of the machines was done based on the designs developed by partners CTEC, CENTI, KLO, DGA and MAR. Partner CTEC designed and constructed two of the pilots: the batch and the R2R sono-machines, while the spray coating machine was constructed by partner MAR with the assistance of CENTI and a subcontractor. The end users KLO, DGA and MAR prepared the facilities and integrated the new machines in their industrial premises. The automation systems on the batch and R2R sono-machines were developed and installed by ICONE. The machines, constructed by CTEC, after coupling with the automation system of ICONE, were checked for their performance prior the delivery to KLO and DGA. In the current stage, all 3 pilots are installed at the industrial premises of the end users.

D3.1 Report and demonstration of real time monitoring by the PBI of the NPs size, concentration and coating uniformity of the end products


PROTECT is building a versatile platform of three pre-commercial manufacturing lines (Technology Readiness Level (TRL) 7) offering antimicrobial, anti-biofilm and biocompatible coating solutions specifically suited to each application area, manufacturer and end user represented in the consortium.
Three existing pilots (TRL 4-5) developed under the FP7 projects SONO and NOVO and with private investment form the basis of PROTECT technological platform.
PROTECT aims to implement methods and/or instrumentation with real time characterisation for measurement, analysis and monitoring at the nanoscale to characterise relevant materials, process properties and product features for ‘real-time nanoscale characterisation’ to ensure ‘reproducibility’ and ‘quality’ of the nano-coated products.


D3.3 Report on the smart metabolic sensing for functional life time monitoring of the end products


This report details the progress in the development of the smart metabolic sensing molecules and their integration in the textiles employed in the PROTECT project. The report is divided into three main section: (i) selection of the sensing probe, (ii) methodologies for implementation in the textiles and (iii) evaluation of smart textile function, i.e. antibacterial capacity and bacterial sensing. In the first selection, electrochromic molecules were chosen for changing color in contact with living bacteria. A number of electrochromic molecules have been tested in solution to evaluate their metabolic sensing capacity and chromatic change. Those presenting most intense colour change in the presence of living bacteria where selected for implementation, i.e. Presto Blue and Prussian Blue. In the implementation in the textile, two strategies have been studied, concretely sonication and cyanotyping, this second only in the case of Prussian Blue. Cotton and cotton-polyester  with and without antibacterial nanoparticles have been used. Initial validation assays demonstrated that: (i) all samples presented sensitivity to the presence of live bacteria, although required long incubation times (≥20 hours for bacterial concentrations above 106 CFU/mL) and (ii) the presence of the sensing probe did not influence in the bactericidal capacity of the textiles also incorporating antibacterial nanoparticles.


D4.1 Report on the hazard determinants for the environment safety profile of the new processes and end products 


This report is mainly related to the establishment of the most representative exposure scenarios and the potential hazards coming from the nanoparticles (NPs) potentially released into the environment during the production and use of the developed materials. The exposure to NPs eventually released by antibacterial coated textiles and water membranes is estimated to occur mainly via inhalation in workers, while for the end-users of textiles the main exposure route is represented by skin contact. Especially in indoor environments, human exposure to the antibacterial NPs may also occur through inhalation after textile abrasion and wearing. Regarding the water depuration membranes, NPs (or other toxic by-products) may be released after leaching, finally impacting on freshwater living organisms. The methods to characterize the NPs environmental release have been set up, mainly following the OECD documents, reporting the test guidelines to measure NPs in workplaces and in the aquatic environment respectively. The bio-assays to screen the toxicity and the hazard determinants for the several new nanomaterials (NMs)proposed as efficient nano-antibacterials have been also set up. The NMs under scrutiny are sonochemically synthesized metal oxides, hybrids, polymers and colloidal solutions of polymers-metal. Such NMs represent almost completely new formulations and no data are available on their toxicity. For these reasons, different toxicity assays representative of the different exposure scenario has been performed, starting with standard in vitro tests on human lung cell lines, and 3D biological models, like human reconstructed epidermis, to go on with freshwater organisms (e.g. zebrafish embryos).

By the described methods, main determinants that may contribute to the hazard of antibacterial coating materials and technologies toward human and environmental organisms were defined.

D6.1 Report on the LCC and LCA of the three coating processes and recommendation for improvements based on inputs from the existing pilots


The deliverable 6.1 is the report accounting for the work done in the first eighteen months of the PROTECT project. The task is specifically referred to the sustainability issues of the identified process technologies for the PROTECT targeted applications and products. The activities planned in Task 6.1 were primarily concentrated to set-up the sustainability assessment model and the computational framework by specifying: parameters, criteria, scope and goal of the analysis. This is the basis of the PROTECT LCA-LCC studies that are being carried out throughout the whole project development: from RTD pilots scaling-up activities with the consequential LCA, to final the scaled-up pilots assessment with attributional LCA. The first impact studies were carried out and accomplished within the framework of the consequential LCA by analyzing possible processing scale-up scenarios. This provides preliminary indications towards optimization and sustainable development of the final pilot units and processes. Specific focus was given to the roll to roll sonochemical coating and to the spray coating preliminary impact assessments related to the fabric antibacterial treatment for both medical and upholstery applications. First results indicate that the processing speed is a key factor to reduce impacts and the limitation or avoidance-of ethanol would be advisable to further enhance the process sustainability

D7.2 Report on the establishment of the PROTECT PLN including a brochure to detail technologies, services and prices


PROTECT established a Pilot Laboratories Network (PLN) to enable industry end users to select the most appropriate technology for their manufacturing need and target cost vs. performance of their products.
The services to be provided are: test of the coating technologies developed within the project, physico-chemical characterisation of the products, nanosafety tests and training seminars related to the novel in situ monitoring of processes and products based on Push Broom Imaging technology.
The PLN is established by the research partners: Universitat Politècnica de Catalunya (UPC), Spain, Bar Ilan University (BIU), Israel, Universita’ Degli Studi Di Milano-Bicocca (UNIMIB), Italy, Centitvc – Centro de Nanotecnologia e Materiais Tecnicos Funcionais e Inteligentes Associacao (CENTI), Portugal, Asociación De Investigación De La Industria Textil (AITEX), Spain, and the know-how and technology transfer provider Kessler ProData GmbH (KES), Germany.

D7.3 Plan for using and disseminating the Know-how


The aim of this report is to present the mid version of the dissemination and exploitation plan of the PROTECT project. It presents the actions that will be taken to disseminate the PROTECT project highlights. Different dissemination avenues are described in relation to the stakeholders to be reached out. Dissemination methods range from the Internet website and social media (Facebook) to journals and conferences. The project website is recognised as a key dissemination medium which brings together all other types of dissemination. Exploitation plan starts with brief explanation of legal boundaries. We propose a set of exploitation tools and identify potential customers of PROTECT solutions.

D7.6 Staff exchange plan between SMEs and R&D centers


Novel technologies as sonochemical continuous technology, spray coating continuous technology and sonochemical batch-mode technology need more skilled employees in the organizations. In this case, staff exchange between SMEs and RTD centers will be organized to facilitate the technology transfer and staff training.
This deliverable establishes linkages and defines the procedures for a successful staff interchange as well as a plan with verifiable activities and timetable oriented to improve the knowledge if the personnel which will participate of the exchange.


D7.13 First business plan for the four business cases


The first Business Plan of PROTECT project includes  a detailed business plan for each of the following antibacterial products to be developed within the project: textile for bed linen, water membranes, catheters and upholstery curtains. These products will be the demonstrators of the technologies developed up to pre-commercial level of coating antibacterial nanoparticles. The technologies are : 2D spray coating based on ultrasound nozzles, 3D ultrasound based  continuous and batch coating.costing.


D7.14 Second business plan for the four business cases


A Second Business Plan of PROTECT project includes integrations and correction to the previous BP (deliverable 7.13) and reports new Business Models and Financial Plans for the four proposed solutions: textile for bed linen, water membranes, catheters and upholstery curtains. Other scenario have been considered based on different production capacity and matching of functionalization unit capacity with substrate production.
A new business model based on servicing external to consortium partners is considered.


D7.15 Deliverable name: Third business plans for the business cases – the four products


According to the World Health Organization, Antimicrobial resistance (AMR) is a growing threat to global health, and for the attainment of the Sustainable Development Goals (SDGs). As a result of infection with drug-resistant bacteria an estimated 700 000 people die each year worldwide. PROTECT project target is to contribute to the eliminate or at least reduce this serous health threat by developing four different antimicrobial products thank to a innovative an unique coating, realized with three different process: Water Membranes, Urinary Catheters, Hospital Linen and public area upholstery. In this third Business plan challenges, specific market, competitor for the 4 products have been deeply investigated, while financial plan and business model have been updated according to the project development, which allowed to an estimation of cost of the spray coating process, as well as more detailed and optimized estimation for R2R and batch processes


D8.2 Quality Control Plan


In order to assure a high quality of the project management, the following tools are being used:

  • A reporting procedure will be applied as described in section 4.
  • There will be defined ways of communication within the consortium and with the EC (Section 4).
  • Deliverables as mentioned in Annex I of the Contract will be delivered as per the attached format and according to the procedure described in section 4 paragraph 4.3
  • Periodical Meetings will be established and carried out according to a defined procedure as described in section 5.
  • The Consortium Agreement, signed by all partners, will direct all consortium related issues e.g. the IPR and financial payments. A short summary of the most important issues settled by the Consortium Agreement is in section 6.

The project defined a well-structured managing process described in section 7.