Work Plan

The work plan has a total duration of 36 months and is structured into 9 different Work Packages (WP), 6 of which on research and technological development, in order to meet the project objectives effectively, which will be monitored and evaluated under management tools (WP1). The proteic and ceramic building blocks will be produced in parallel work-packages (WP2 and WP3, respectively) and further combined to render biodegradable and bioresorbable 3D printing inks based in collagen or composite formulations (WP4). These inks will be used to produce scaffolds by 3D printing (WP4), which performance regarding bone tissue engineering will be assessed by in vitro and in vivo assays (WP5). To assure a sustainable production platform of proteic materials (collagen and crosslinker), a work-package dedicated to marine sponge farming was designed (WP6). The integration of the planned technology in a clinically relevant approach will be addressed by establishing a methodology to build 3D CAD models for 3D printing of scaffolds, based on reverse engineering from clinical patient CT scans (WP7). Regulatory, IP and dissemination issues will be the subject of two fully dedicated work-packages (WP8 and WP9, respectively), mandatory to establish the interaction with relevant stakeholders towards the development of clinically relevant products and processes, which can later be explored as market products or services offered by the enrolled SME, spin-off to be created and selected allies.

The overview of the WPs is depicted below.

WP1: Management: It is essential to create an efficient organisational and coordination structure to direct and control the project running, which will ensure the optimal performance of the consortium. The responsibilities of each partner within each one of the WPs are clearly identified and each WP will have a leader, included in a project management structure headed by the project coordinator Tiago H. Silva (UMinho). He has the responsibility for the direction and control of the project, interaction with the MERA.NET managing bodies and will make the link between the different partners involved.

WP2: Extraction, purification and characterization of the collagen and crosslinker-factor: The crosslinker-factor, a low hydrophilic and slightly acidic protein, will be extracted according to Fassini et al. [7] and further purified using chromatographic techniques. Collagen Type I will be extracted from marine sponges according to Fassini et al. [7] and from salmon skins using a standard acidic treatment.

WP3: Preparation of ionic-doped calcium phosphates nanopowders: CaPs will be isolated from salmon fish bones by calcination and then doped with different ions. Alternatively, well-established synthetic doped CaPs will be produced via aqueous precipitation from precursors of Ca and P, and precursors of the doping elements (Sr, Zn, and/or Mn), in a medium of controlled pH, followed by heat treatment. CaP nanopowders will be produced by grounding and wet attrition milling.

WP4: Optimizing materials and 3D-printing: Different formulations of collagen(s)/crosslinker/doped CaPs will be prepared and evaluated to mimic the characteristics of natural bone, addressing the rheological properties requested to allow plottability. Further, bone tissue engineering scaffolds will be produced by 3D printing, based on CAD models, and processing parameters will be changed while evaluating the printing resolution and fidelity (similarity between CAD model and microCT scan of the produced scaffold).

WP 5: Evaluation of biological performance: In vitro bioactivity studies in simulated body fluid, degradation behaviour by incubation in enzymatic media, release profile of ionic-dopants, in vitro biological performance firstly using osteoblast-like cell lines (SaOS2) and then human stem cells, as well as in vivo performance in mice or rabbit animal models will be performed, following methodologies well established on the partners’ facilities.

WP 6: Sponge farming: An integrated multitrophic aquaculture system will be implemented combining fish farming with the marine sponge C. reniformis. This sponge farming approach will not only represent a sustainable production platform of sponge biomass and subsequently of collagen and crosslinker, but also represent a strategy to mitigate the effects of sea fish farming (bioremediation).

WP 7: Establishing a protocol from data acquisition to bone TE scaffolds printing: A reverse engineering strategy will be adopted in order to establish a protocol that will make possible the translation from data acquired with Computed Axial Tomography to the 3D-design of the scaffold in a CAD software. In particular a new strategy must be established when the aim is to produce patient case-specific bone TE scaffolds.

WP 8: Regulatory aspects, certification and IP protection/technology transfer: This WP will be devoted to the development and effective implementation of the instruments to assure a complete protection of potentially exploitable results. A pre-market strategy both in short and long term, as well as the basis for the dissemination of knowledge and regulation of patent filling and technology transfer will be defined. Patent filling and product commercialization are also an end-goal of this general WP.

WP9: Dissemination: Considerable effort will be directed towards building a solid communicational basis. Emphasis will be put on conveying pertinent information to the academic and research community, as well as the industry, hospital and patient Associations via several routes (e.g., newsletters, social networks, scientific manuscripts, communication at conferences, etc). Interactive sessions during project meetings and workshops will be also considered.