Periodic Reporting for period 2 - RESTORE (User-centred smart nanobiomaterial-based 3D matrices for chondral repair) Reporting period: 2020-07-01 to 2021-12-31 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 During the first 36 months, the RESTORE consortium focused on producing the smart 3D matrices able to repair cartilage lesions. The main results achieved are:I. Standard Operating Procedures (SOPs) for RESTORE nanobiomaterial safety assessment were developed and performed for the selected nanoenabled Col-PLA scaffolds (in vitro and in vivo safety assessments). Work on establishing the regulatory route for the 3D matrix, COPLA® is ongoing.II. Different groups of nanomaterials were produced/selected and characterised following the established SOPs and their bioactivity assessed in vitro : a) nanoparticles that can carry pharmacologically active drugs to fight undesirable threats such as inflammation cartilage tissue degradation and infection and b) nanomaterials (stimuli-responsive nanomaterials) that are able to activate cartilage cells via remote stimulation. The best nanomaterials formulations were incorporated in Col-PLA scaffolds. The bioactivity of the obtained nanoenabled scaffolds was performed and the most efficient were pre-selected for in vivo efficacy testing.III. The Incorporation of different formulations of nanomaterials in is ongoing. IV. The prototypes of platforms where the stimuli-responsive nanomaterials can be tested in vitro with cartilage cells were designed, improved and have advanced to the in vitro studies. V. The bioink that will serve for cartilage tissue bioprinting was further improved to make it more suitable for cartilage cells.VI. The bioprinting platform for human cartilage printing is further improved by upgrading print heads and the software.VII. The beta version of the patient-specific anatomical models for condyle lesion database was created and is online and available to the public.The dissemination and communication activities of the project have also been carried out. For the first reporting period, we aimed to create project visibility and disseminate the project among academic researchers in nanomedicine, biomaterials and regenerative medicine, the orthopaedic community as well as the general public. Further connection between the website and the social platforms was encouraged.The RESTORE website (https://restoreproject.eu/) had 11928 views, 3934 visitors and 54 posts, fulfilling the yearly goals. A second version of the website was launched on M15 and a Twitter and a Facebook account were launched at M16.RESTORE scientific results have been presented in several scientific events such as congresses, workshops, displays/fairs, etc. As planned, 1 press release was published at the beginning of the project. 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) Progress beyond the state of the art:During the first 36 months of the project, the following progress beyond the state of the art was achieved:I. The best formulation of nano enabled Col-PLA scaffold has been selected based on in vitro efficiency studies, in vitro. In vivo validations are ongoing. II. The prototypes for the in vitro stimulation testing platform were improved and are being tested.III. The beta version of the database of patient-specific anatomical models for condyle lesions has been uploaded and is online and available to the public ( https://restore-project.ru.is/index )VI. Reliable data on the safety of almost all of RESTORE’s 3D matrices building blocks were obtained.Expected results until the end of the project:I. Functional nanocarriers capable of mitigating inflammation, inhibit cartilage ECM degradation and hinder bacterial growth.II. Stimuli-responsive nanobiomaterial tuned to physical stimulus.III. Viable 3D bioprinted human cartilage microtissues from patient-specific anatomical models.IV. Nanoenabled 3D matrices for efficient chondral lesion repair.V. Device to stimulate and monitor cartilage regeneration.VI. Reliable data on the safety of RESTORE’s nanoenabled solutions.Potential impact:Patients suffering from knee chondral lesions have reduced quality of life. By using the solutions developed in RESTORE, patients either young or aged, can be rehabilitated and regain an active life in shorter times (reducing the absenteeism of working patients). Relative to traditional approaches (chemical and biologic), cell-based products combined with nanocarriers are considered to be expensive to manufacture. However, RESTORE tailored nanocarriers hold great potential to reduce treatment's costs, as the dosage of bioactive molecules will decrease from milligrams to nanograms, with less side effects. The stimuli-responsive nanobiomaterials do not represent a significant increase in the final nanoenabled solution, since their manufacturing costs are low. The integration of the developed smart nanobiomaterials to the 3D matrices is planned with regard to minimising additional manufacturing costs and, therefore, providing an affordable nanoenabled solution. Its application by orthopaedic surgeons in hospitals or specialised clinics is expected to improve patient quality of life and will ease the burden from the social and health systems and from the families and societies. Finally, the use of SOPs accepted by regulatory bodies from an early stage of product development will ease the translation from lab to clinic and contribute to results directly useable in regulatory documentation at a later stage. Also, the focus on sterile production processes by continuous sterility and endotoxin testing will contribute to an accelerated scale-up production and translation from lab to product. RESTORE Website - https://restoreproject.eu/
