Periodic Reporting for period 3 - RESTORE (User-centred smart nanobiomaterial-based 3D matrices for chondral repair) Reporting period: 2022-01-01 to 2022-08-31 Summary of the context and overall objectives of the project Injury to articular cartilage is recognised as a cause of significant joint morbidity. The initially small focal chondral lesions gradually increase not only in circumference (perimeter) but also in length and commonly result in the development of osteoarthritis (OA), eventually leading to progressive total joint destruction. The most recent report of the Osteoarthritis Research Society International estimated that OA affects 242 million people globally. In Europe, it has been reported that the average total annual costs of OA per patient ranges from €1,330 to €10,4522. These facts clearly support the need for efforts to take on the challenge to repair even minor articular cartilage injuries or lesions. The RESTORE project addresses this clinical need by developing an effective approach to treat knee chondral lesions. The RESTORE concept is to realize a new generation of smart nanoenabled 3D matrices able to fit into complex lesion geometry, modulate undesirable biological events, and remotely control, stimulate and monitor cartilage repair. To create smart and functional 3D matrices, RESTORE is developing (i) nanobiomaterial-based nanocarriers to modulate undesirable biological threats, (ii) stimuli-responsive nanobiomaterials tuned via external physical stimulus, and (iii) non-invasive wearable and wireless device for remote stimulus and regeneration monitoring. The nanobiomaterials are planned to be integrated into cutting-edge 3D matrices that meet the needs of severe and mild chondral lesions. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far RESTORE’S MAIN ACHIEVEMENTS:1. Standard Operating Procedures (SOPs) for the safety assessment of RESTORE nanobiomaterials were established. Safety assessments following these SOPs were performed for the developed nanomaterials and nanoenabled scaffolds (in vitro and in vivo safety assessments). The use of SOPs accepted by regulatory bodies from an early stage of product development will ease the translation from laboratory to clinic and contribute to results directly usable in regulatory documentation at a later stage.2. Different groups of nanomaterials were produced/selected: a) polymeric nanoparticles that can carry pharmacologically active drugs to fight undesirable threats to regeneration and b) nanomaterials (stimuli-responsive nanomaterials) that are able to activate cartilage cells via remote stimulation. The nanomaterials were characterised following the established SOPs and their bioactivity was assessed in vitro. The best nanomaterial formulations were incorporated in the scaffolds or in the designed bioinks for human cartilage bioprinting.3. The integration methodologies for the nanocarriers and the stimuli-responsive nanomaterials into either cutting-edge 3D matrices (COPLA® and Col-PLA scaffolds) were successfully achieved. The COPLA® Scaffold, already used in the veterinary market, is a registered trademark of the RESTORE partner Askel Healthcare. The Col-PLA scaffold is partly produced by Askel Healthcare and partly at INEB following the COPLA® Scaffold production protocol provided by Askel Healthcare after a Non-Disclosure Agreement (NDA) with INEB.4. The bioactivity study of the obtained nanoenabled scaffolds was assessed in vitro and the most efficient scaffolds were selected for in vivo efficacy testing in small animal models. The effectiveness of the nanoenabled matrices in preventing microbial infections was performed in the induced tissue infection mice models while the mitigation of inflammation and inhibition of cartilage extracellular matrix degradation was investigated in a rabbit osteochondral lesion model.5. The prototypes of platforms where the stimuli-responsive nanomaterials can be tested in vitro with cartilage cells were designed, improved, and have advanced to a level for use in in vitro studies. 6. Polymer-based Bioink, previously tested for cell bioprinting, was further improved by biofunctionalization to make it more suitable for the bioprinting of human cartilage cell. In addition, the bioink was nanoenabled with the nanomaterials developed in the project. The printability of the different bioink formulations was assessed. 7. Protocols for 3D bioprinting cartilage microtissues using nanoenabled bioinks combined with human chondrocytes derived from iPSCs were established and validated.8. The bioprinting platform of the partner BRINTER was improved by upgrading print heads and the software.9. The patient-specific anatomical models for a condyle lesion database were created and this database is online and available to the public: https://restore-project.ru.is/index.The dissemination and communication activities of the project have also been carried out via website, the different platforms (linked: LinkedIn (https://www.linkedin.com/in/restore-project-1b9701201/ Facebook: https://www.facebook.com/profile.php?id=100048414296631 and Twitter: https://twitter.com/RESTOREProject3) several scientific events such as congresses, workshops, displays/fairs, etc Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) Examples of RESTORE Impact by the end of the project:-The RESTORE project has led to 2 patent applications on the use of nanocarriers with multiple drug release and nanoenabled matrices and 1 patent application on bioprinting technology for biological tissues, including human cartilage.-During the project, partner Askel Healthcare has further developed COPLA® towards human use and achieved the breakthrough designation for medical devices from the FDA Breakthrough Devices program. With this, the discussions with FDA have started and the plans for clinical studies are ongoing for future marketing authorizations. The integration of nanoparticles in Col-PLA scaffold showed encouraging results that will open interesting future clinical applications. BRINTER has developed and tested new systems for printing various bioink combinations. This development will facilitate the journey towards significantly improving cartilage bioprinting technology (as well as other tissue printing) and make the technology commercially viable.-The nanocarriers have been successfully characterised according to EUNCL and ISO-related protocols in terms of their physical-chemical properties, safety, biocompatibility, and bioactivity. Thus, the RESTORE project has brought these nanocarriers closer to human trials. Moreover, their incorporation into the scaffold and their extensive characterisation according to regulations offer an innovative solution for articular chondral repair. The work carried out in the RESTORE project will allow to have a specific candidate with a defined formulation and established quality control that will facilitate the pre-clinical regulatory studies required to finalize the product Dossier and bring the 3D matrices closer to the patient, thus to the market. RESTORE Website - https://restoreproject.eu/
