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Unlocking the research potential of 3Bs Group, University of Minho, in Nanomedicine field to strengthen its competitive position at the European level

Final Report Summary - POLARIS (Unlocking the research potential of 3Bs Group, University of Minho, in Nanomedicine field to strengthen its competitive position at the European level)

Executive Summary:
3Bs has been a lighthouse and an example on the exploitation of the regional research potential within the biomedical area. Its infrastructures and knowledge focus the macro and microscale processing of biomaterials. However, new advancements in the field are based on the nanostructuring of biomaterials in order to achieve enhanced diagnosis, treatment and prevention of diseases and traumatic injuries (NanoMedicine). Thus, POLARIS is created to strengthen the 3Bs position in the field of biomaterials by supporting 4 nanotech-based research lines (RLs) relevant within NanoMedicine, namely: 1) Supramolecular chemistry and self-assembling methodologies; 2) Biosensors, bioarrays and microfluidics; 3) Biofunctionalisation of nanoparticles; 4) Top-down nanofabrication and surface characterization tools. A set of instruments relevant to these RLs will be acquired: AFM; lithographic equipment; and electrokinetic analyser. An upgrade of 3Bs clean room is also supported. Additionally, experienced researchers and technical staff are to be recruited to: act in the described RLs; operate the state of the art equipments; and to implement and manage a well-structured Intellectual Property (IP) and innovation system. Interactions of 3Bs with European partners of Excellence are promoted (i.e. two-way secondments) to enhance its scientific outputs. Two workshops, one training course and one conference will contribute to: improve the 3Bs knowledge on the nanotech manipulation of materials and their interaction with cells; enhance the involvement of 3Bs members in IP management and protection; and strengthen the dissemination and exchange of knowledge within the Nanomedical research community. This strategic investment will: promote a brain gain strategy; enhance 3Bs IP protection and innovation standards; confer to 3Bs the necessary tools to improve its competitive position at the European level, both in terms of successful funding and scientific outputs.

Project Context and Objectives:
The POLARIS Action Plan was designed to confer to 3Bs appropriate expertise, knowledge, infrastructures and equipments in the field NanoMedicine. Additionally, POLARIS includes a set of measures to improve the group’s dissemination, IP and innovation strategies allowing it to consolidate its Excellence in the field and to better exploit the commercial potential of its developments.
In a scientific perspective, POLARIS consolidates 4 NanoMedicine-related Research Lines (RLs): RL1 - Supramolecular chemistry and self-assembling; RL 2 - Biosensors, bioarrays and microfluidics; RL3 - Biofunctionaliztion of nanoparticles; and RL4 - Nanofabrication and characterization. These RLs are already established in the group, although, most of the present research developments are in the micrometer range, limiting the manipulation of the materials and the cell-biomaterial interactions. Some incursions in the nanometer range has been achieved, most of them with interaction with international institutions of Excellence (from USA, Japan and Europe).
This set of 4 RLs is the basis of the 3Bs strategy to consolidate its competitive position within the field of biomaterials (its main area of expertise). These RLs are all within the field of NanoMedicine and they have demonstrated their potential to: improve the scientific and technological Excellence of 3Bs; explore new scientific frontiers that are unattainable with the manipulation of the materials features at the micro-scale. In order to better exploit their research potential, POLARIS supports the recruitment of experienced researchers for the 4 RLs. This measure intends to confer to 3Bs appropriate expertise to better cope with the demands of this top-level research domain.
The manipulation of biomaterials at the nanoscale demands for specific equipments and infrastructures. In this perspective, the 3Bs group has been joining efforts to acquire specific equipments that are critical in the processing and characterization of nanostructured biomaterials and nanobiomaterials, e.g. Electrospinning, Dynamic Light Scattering (DLS) and Ellipsometry. All the available equipments are relevant to the field of NanoMedicine, although, in order to execute Excellent research at the nanoscale it is also mandatory the use of additional state of the art equipments not available at 3Bs, nor at the University of Minho. POLARIS addresses this 3Bs limitation acquiring: surface characterization equipments, such as, an Atomic Force Microscope (AFM) and an ElectroKinetic Analyser (EKA); and nanoprocessing instruments, such as a nanolithographic equipment. The operation of these 3 equipments demands for a full time technician with expertise and knowledge to manipulate them. POLARIS contributes to this necessity through the recruitment of experienced technicians that will be responsible for the correct operation of the newly acquired equipments.
Some of these equipments also necessitate the adaptation of a clean room to reduce the number of particles in the air. This last adaptation is critical when nanoscale surface modification or patterning is performed. When these methodologies are applied there is a high probability of creating distortions on the patterning/modification if surface contaminations (e.g. dust) are present.
The implementation of the previously outlined RLs at 3Bs is strengthen through the exchange of knowledge between 3Bs and the partnering institutions (Chalmers, MPG, USTRATH and NUID UCD). A team composed by 3Bs permanent staff and contracted researchers are assigned to each RL. Within these teams a set of researchers will be selected for secondment to the partner institution that possesses the expertise able to enhance the RL research output. Additionally, the contracted 3Bs experienced technician will be seconded to the partners with expertise on the use of AFM, lithography and EKA, where she/he will learn how to manipulate the instruments in order to execute the nanoscale processing and characterization needed under each of the described RLs. Moreover the secondment of experienced researchers from the partners to 3Bs will be also considered in order to give in loco support to the RLs.
The workshop and conference tools are projected under POLARIS to improve research excellence within each of the RLs. POLARIS is also designed to enhance the 3Bs dissemination strategy using different means to reach the general public and scientific community as a way of increasing their awareness to the Nanomedical solutions developed by the group and the scientific community in general.
Finally, the 3Bs group is already aware of the importance of the commercial exploitation of its scientific developments, although, one of POLARIS measure intends to strengthen this perspective within the group. In this sense, it is important the recruitment of an experienced Intellectual Property (IP) and Innovation manager. This manager will, in coordination with the 3Bs director (Prof. Rui L. Reis), establish a well-defined strategy to improve the 3Bs commercial outputs.

Project Results:
WP1 Recruitment of experienced researchers and technical members
With the support of POLARIS we were also able to maintain the number of experienced researchers actively working in the Nanomedicine field. Some of the contracted researchers applied to the National program “FCT Investigador” and, in 2 cases they were successful, securing the financing for their working contract for more 5 years (beyond the timeframe of POLARIS). Other researchers that have contributed to POLARIS, but were not contracted directly by the project were also successful in these national programs, enabling the strengthening of the research team that works at 3Bs / University of Minho in the field of Nanomedicine.
WP2 Acquisition of equipment and improvement of infrastructures
We acquired the projected set of larger equipments, i.e. atomic force microscope (AFM), electrokinetic analyser (EKA) and the electron beam lithographic equipment/scanning electron microscope (SEM/FIB).
During the execution of the project it was identified a series of additional medium-size equipments that would significantly enhance the capacity of the 3Bs Research Group / University of Minho to execute high-level scientific research in the Nanomedicine field. In this context, we suggested to reallocate part of the POLARIS budget to the acquisition of these equipments, maintaining the whole acquisition of equipments below the 45% limit on the project budget. Upon approval by the project officer (by e-mail on the 20th of February 2015), we initiated the process of acquiring: Mass spectrometer (MS); Quartz Crystal Microbalance with Dissipation (QCM-D); X-Ray Fluorescence (XRF) detector for the scanning electron microscope (SEM); Spectrofluorometer; Circular Dichroism (CD); Fluorescence module for the atomic force microscope (BioAFM); Photolithographic equipment; Sputter coater. These acquisitions were executed by direct invitation to the supplier to present a proposal. All the contacted companies were able to provide the equipments that we requested and the whole process of delivery and installation finished by the end of September 2015.
During the project we were also able to upgrade the clean room facility, where some of the equipments (acquired under POLARIS) are already installed and under operation (e.g. photolithographic instrument).
The whole set of equipments acquired under POLARIS was extremely valuable and capacitated the 3Bs Research Group / University of Minho to execute research at the nanoscale, namely in the field of Nanomedicine, as detailed below through the success indicators.
WP3 Exchange of knowledge and experience through secondments
We were able to enhance our knowledge and experience through the exchange of knowledge with our collaborators in the field of Nanomedicine. Two of the most relevant were the ones that were established with the Max Planck Institute for Intelligent Systems and with the University of Strathclyde, with relevant scientific outputs (published in a series of high impact journals, e.g. Journal of the American Chemical Society). The remaining part of the exchange of knowledge was also executed with the restrictions expressed in the deliverables D3.1 D3.2 D3.3 and D3.4. The execution of the incoming staff exchange was approximately 70%. A series of research tasks were defined and are projected to continue beyond the project timeframe. In particular the collaborations with the University of Strathclyde (Prof. Rein V. Ulijn) and the Max Planck Institute for Intelligent Systems (Prof. Joachim Spatz) are ongoing and new papers are projected to be submitted in the near future. It is relevant to point out that, although some of the projected experiments present some ethical issues (mainly derived from the fact that we use adipose-derived stem cells from human origin) the ethical protocols, that were the base support at the time of the signature of the grant agreement, are still valid. All the biological experiments (that present ethical issues) continue to be executed under the guidelines defined and detailed in the annex 1 of the grant agreement.
WP4 Organization of workshops, training course and conference
We organized a series of 4 events. The 1st POLARIS workshop was organized in Porto (at Porto Palácio Hotel and Spa) being a great success, as detailed in the deliverables D4.1. The topic of this event was “Nanosystems for Medicine: fundamentals, synthesis and applications” with the participation of 9 invited speakers and an audience of 51 attendees (deliverable 4.1). The 2nd workshop entitled “Top-down fabrication and nanocharacterization techniques to develop nanosystems for biomedical applications” and the POLARIS conference (“Challenges in Nanomedicine and Regenerative Medicine”) was also organized with great success, as detailed in the deliverables D4.2 and D4.4. In the case of the 2nd workshop, we counted with the participation of 10 invited speakers and an audience of 41 attendees. In the case of the POLARIS conference, participated 8 invited speakers and 71 attendees. The 3Bs group also organized the IP training course at our facilities (deliverable D4.3). In this event, we hosted 11 invited speakers and 79 attendees. These 3 events added to the 1st workshop (that occurred during the previous reporting period) completed the execution of the events scheduled under POLARIS.
WP5 IP and Innovation strategies
An IP and Innovation manager was also contracted and defined a 3Bs IP strategy (reported under deliverable 5.1). This strategy was defined based on an initial evaluation of all the patents created by 3Bs, namely considering their exploitation stage and potential to be industrially exploited. During this evaluation it was noticed that same of the patents were not valid because of several bureaucratic issues, e.g. deadlines that were not met, leading to the loss of protection. All this information was used to create a methodology that significantly reduces these flaws and that creates a better pipeline of protected developments that can actually be transferred to the industrial sector bringing added value to the group. An innovation strategy was also established and reported under deliverable D5.3. The IP and Innovation manager also was actively involved in more recent patent applications from the 3Bs Research Group. At the moment all the patent applications submitted by 3Bs are passing through this office, as detailed in the report of activities (deliverable D5.4 and D5.6).
WP6 Dissemination of knowledge
It was possible to publish a series of papers in the field of Nanomedicine, as detailed in the report of activities for the first (D1.2) second (D1.3) and third (D1.4) year of the project. The most relevant topics in each research line were:
• Synthesis of Amyloid-beta (Aβ-42) peptide (RL1)
• Influence of polyelectrolyte molecular weight on layer-by-layer film construction and CD44 receptor binding (RL1)
• Tunable nano-carriers from glycosaminoglycan block copolymers (RL1)
• Design and synthesis of bioactive self-assembling peptides (RL1)
• Interaction of CD44 cell receptor with nanostructured multilayered films of hyaluronic acid and poly-L-lysine (RL1)
• Natural polyphenols as inhibitors of the self-assembling of amyloid-beta 1-42 (RL1)
• Behaviour of adipose stem cell on glycosaminoglycans platform with protein (RL2)
• Phosphorylated carbohydrate amphiphile to selectively target and kill osteosarcoma cells (RL2)
• Fractionation of sulphated glycosaminoglycans of different molecular weights (RL2)
• Modification and characterization of gold thin films with a thiolated GAG (RL2)
• Influence of bilayered silica nanoparticles over the osteogenic differentiation of human mesenchymal stem cells (RL3)
• Development of magnetic nanocomposites targeting musculoskeletal regeneration (RL3)
• Stimulating hBMSCs to secrete EVs during culture (RL3)
• Magnetic nanoparticles to guide tissue regeneration (RL3)
• Gold nanoparticles functionalized with hyaluronic acid-terminated self-assembling monolayers (RL3)
• Synthesis of acemannan and aloe-emodin nanoparticles and their characterization (RL3)
• Production of chitosan nanofibers (RL4)
• Gellan gum-hydroxyapatite composite hydrogels for bone tissue engineering (RL4)
• Chitosan/nanohydroxyapatite composites as scaffolds for bone tissue regeneration (RL4)
• Development of marine biopolymer based nanoparticles for drug delivery systems (RL4)
• Fabrication and characterization of synthetic membranes with biochemical and topographic cues (RL4)
• Development of antibacterial surfaces bioinspired on the micro/nanotopography of natural surfaces (RL4)
• Development of peptide-containing systems that inhibit angiogenesis (RL4)
• Magnetically actuated SPCL membranes for tissue engineering (RL4)
• Production of electrospun membranes using marine origin collagen (RL4)
• Fabrication of multilayer scaffolds for bone tissue engineering (RL4)
Most of the work is being prepared for submission in scientific journals. The ones that, upon evaluation by the IP and Innovation manager, are considered to present a significant industrial and commercial relevance might be considered for patenting.
WP7 Project management
We achieved all the main objectives for the period and for the overall projects. The deviations that were introduced with the agreement from the project office were related with the acquisition of additional middle-sized equipment relevant for the field of Nanomedicine. All these tasks were successfully executed, as can be evaluated by the images of the instruments installed in the 3Bs labs. In addition, we were able to significantly execute the exchange of staff. The events were completed as scheduled.

Potential Impact:
As detailed and presented in the 2nd periodic report it is clear that the POLARIS project was able to improve the 3Bs infrastructures in order to interact better with the region and their institutions. In fact, the number of requests from external institutions (i.e. academic and industrial) for services using the type of equipments acquired under POLARIS increased exponentially, from less than 10 per year to close to 100. It also emphasises the higher relevance of the Nanomedicine area in the PhDs ongoing in the group. In this topic the percentage of Nanomedicine-related ongoing PhDs increased from approximately 15% to approximately 35%. While before POLARIS the group was publishing between 60 and 70 papers per year; after POLARIS this indicator increased to between 90 and 100. The average impact factor of the journals where 3Bs publishes it research work also increased: the average impact factor of the journals where 3Bs publishes its 10 most relevant papers per year changed from ~8 to above 25. The improvement of the capacity of 3Bs to attract scientific funding is also clear: it almost doubled after the starting of POLARIS, passing from the 1.5-2.0M€ mark to above 3.5M€ per year. In addition, it is clear that POLARIS produced a significant impact in the number of applications to the H2020 calls, where the total budget at the submission stage significantly increased with the POLARIS project (changing from close to 1M€ to close to 2.5M€).
Given this data it is clear that the POLARIS project was able to improve the 3Bs infrastructures in order to interact better with the region and their institutions. It also emphasises the higher relevance of the Nanomedicine area in the PhDs ongoing in the group, while the number and quality of the papers published by the 3Bs members also increased. The capacity of 3Bs to attract scientific funding is also clear, it almost doubled after the starting of POLARIS. In addition, it also increased the total budget of the proposals (with 3Bs involvement) submitted to the FP7/H2020 (before evaluation).

List of Websites:
http://projectpolaris.eu/EN/