The First Year of GreenNanoBone: Laying the Foundations for a New Approach to MRONJ Prevention and Treatment
Since the official kick-off meeting in Gdańsk on 30 June 2024, the GreenNanoBone consortium has spent its first project year building the scientific, technological and regulatory basis for an entirely new strategy to prevent and treat Medication-Related Osteonecrosis of the Jaw (MRONJ).
Optimisation of Pectin-based Biomaterial
The GreenNanoBone project focuses on developing sustainable biomaterials for advanced biomedical applications. One of the key components in this effort is the use of plant-derived polysaccharides with tailored structural properties. The University of Copenhagen (UCPH) supplies enzymatic modified rhamnogalacturonan I (RG-I), a pectic polysaccharide, to the project partners. This modification aims to optimise the functional properties of RG-I for incorporation into nanostructured biomaterials. Future work will focus on evaluating the performance of the modified RG-I in composite biomaterials and optimising the scale-up process to ensure cost-effectiveness and sustainability.
The GreenNanoBone Materials
The University of Liverpool is responsible for actually making GreenNanoBone. The team in Liverpool has paved the way towards scalable, GMP-grade manufacturing of injectable, printable microgels incorporating GreenNanoBone’s enzymatically modified RG-I pectins. The team has been busy providing an agile platform to explore GreenNanoBone’s formulation. They have also done stellar work exploring the stability of GreenNanoBone microgels at physiological temperatures, connecting fundamental theory on components and formulation conditions required to make GreenNanoBone stable in patients. Additionally, the team has been working hard on labelling of RG-I, allowing us to integrate it into GreenNanoBone microgels for high-throughput fluorescence microscopy studies that will feed into machine learning work next year.
Preparing the Scale-Up
In this initial project phase, the team at MyBiotech focused on preparing the stage for the eventual scale-up and manufacturing of the 4D-microgels in compliance with regulatory guidelines. In a first, intermediate achievement, a report has been composed with the support of partners UNILIV and UCPH which provides early identification of required quality, efficacy and safety (QES) parameters and documentations for the GreenNanoBone products. After identifying the current status of the development process and critical information gaps, the team is currently transitioning towards more robust and advanced manufacturing protocols including a clear work plan to fill the identified gaps. These will facilitate a smooth technology transfer phase and eventual GMP-compliance in the later phases.
Pre-clinical Testing
An important step on the way to clinical testing of the developed materials is the biosafety tests, which evaluate potential negative effects on healthy cells. In this context the team at the University of Liverpool has made strong early progress in setting up the safety foundation for GreenNanoBone. They began with the RG-I pectin produced by the University of Copenhagen, reviewed its full production record, and confirmed they had everything needed to trace its history and understand where impurities might arise. The primary outcome of this year is that the safety pipeline has been built, tested on real material, and is already providing helpful insight into how these plant-based components behave. During the first year of GreenNanoBone, the Medical University of Gdańsk (MUG) has made substantial progress in establishing the biological and clinical foundations required for the project’s in vitro and ex vivo studies. The team has optimised procedures for obtaining clinical material, including its transfer and transport conditions, ensuring high-quality samples for downstream analyses. In parallel, the team has completed the necessary documentation and secured ethical approval from the Bioethics Committee, enabling continued progress in the upcoming project phases.
Understanding MRONJ
An important aspect of the GreenNanoBone project is to understand the biological mechanisms underlying human MRONJ. To this end, both 2D in vitro models and 3D MRONJ models will be created to examine how medications, such as bisphosphonates negatively affect bone biology, including the inhibition of osteoclast activity, osteogenesis, bone resorption and angiogenesis, factors that contribute to MRONJ development, particularly following dental trauma or surgery. In a first step Tampere University is studying the inhibitory effects of bisphosphonates (BP) on gingival keratinocytes and fibroblasts, as well as on immune cells (macrophages and T cells). Several project partners are working together on these tasks. Preliminary testing with bisphosphonates is ongoing at the University of Bergen (UiB). MUG has initiated the isolation of gingival cells using protocols obtained from UiB. At TAU, immune cells are available in frozen stock, enabling initial immune cell response assays, although the actual experiments will be performed using freshly isolated cells. To develop and validate an in vitro 3D MRONJ model, the University of Aveiro has initiated experiments using the hydrogel-stamping technique. Preparatory work on the MRONJ-on-Chip platform has been initiated at TAU. Aarhus University and Aarhus University Hospital have secured ethical approval for the collection of human samples to investigate the mechanisms underlying MRONJ, and they have optimised protocols for the analysis of both healthy (control group) and MRONJ-affected (disease group) samples. Additionally, a video has been produced to support the recruitment of patients, health professionals, and experts to gather input on the GreenNanoBone project and to raise public awareness of MRONJ.
Laying the Foundation for Social Acceptance of the GreenNanoBone Approach
Amsterdam UMC Foundation has started working on the development of an expert mental model, which will describe the science related to the development and use of GreenNanoBone, perceived positive and potentially negative consequences, and existing uncertainties. In addition, the team interviews experts on potential ethical considerations related to GreenNanoBone. An interview protocol has been developed by studying existing literature on relevant topics such as MRONJ and biological nanomaterials and existing interview protocols in the field of ‘a mental models approach to risk communication’.
The Pathway to Regulatory Acceptance
Asphalion has started to work together with the other members of the consortium on the definition of a "Regulatory classification and roadmap" for GreenNanoBone. Product classification takes into account the known functions and mode of action of the product, and identifies key data and studies needed to support product qualification. At the same time, a regulatory pathway for potential final product classifications is being defined, closely integrated with GreenNanoBone’s broader exploitation and intellectual property (IP) strategy. F6S coordinated the identification and prioritisation of Key Exploitable Results (KERs) to map commercial pathways, market needs, and business opportunities. These efforts underpin the development of the preliminary business plan, designed to map viable commercial pathways and align the project’s value propositions with current market needs. Furthermore, F6S leverages its extensive global network of over 5 million founders and investors to identify strategic partnerships and secure follow-up funding for the project's long-term sustainability. EURICE introduced a consortium-wide framework for IP ownership, protection routes, access rights, and commercialisation procedure. These efforts ensure that GreenNanoBone’s sustainable biomaterials and modelling technologies can progress smoothly from research to market, maximising both scientific and societal impact.