Project Portugal 2030

Summary

The BioNanoTen project aims to developed a nano-tenosome as a responsive nanocarrier to deliver biomolecules with therapeutic value to refine the strategies for guiding and stimulating tendon regeneration through modulation of the inflammatory response to address the unmet needs for a new therapeutic solution to treat tendinopathy with a strong potential to be translated into the clinical setting. To achieve this goal will implement a sophisticated nanocarrier system, integrates potential biomarkers (e.g. microRNA) found in exosomes, and integrated superparamagnetic iron oxide nanoparticles (SPIONs) functionalized with a peptide to inhibit a pro-inflammatory pathway. This innovative system (nano-tenosome) not only allows for the precise customization of biological cargo but also favouring macrophages regenerative phenotype and guiding NFkB-inflammatory pathway towards a molecular tissue-oriented approach. Thus, the objectives of BioNanoTen are: i)Explore the role of hypoxia on the production and the cargo of exosomes from human tendon derived cells (hTDCs) as a therapeutic agents to educate cell behaviour and proceed with the inflammation resolution. The hypoxic conditions maximize exosome yield from the cells and enhance their regenerative potential by increasing the presence of miRNAs with regenerative properties. The miRNA and others (e.g. growth factors, cytokines, enzymes) molecules known to regulate cellular responses crucial for intercellular communication and tendon homeostasis; ii)Develop an innovative nano-tenosome system to ensure transport and delivery of the therapeutic molecules as biological cargo to the inflamed tissue. Thus, BioNanoTen will develop a cell instructive nano-tenosome system produced from exosomes release by tendon cells enriched molecules with therapeutic role, combining with SPIONs to assist the remote monitoring, localization and release of the cell instructive cargo in tendon niches. The nano-tenosome will be designed to carry customized therapeutic cargos to target tendinopathy, and to provide a peptide for targeting an inflammatory signalling pathway (NFkB) present in tendon cells and macrophages towards inflammation resolutions; iii)Confirm the efficacy of the nano-tenosome system in modulating the inflammatory response within a co-culture of hTDCs and macrophages in an artificially induced inflammatory environment. These co-culture models mimic the cellular environment within tendon tissue, providing a platform to assess the nano-tenosome’s ability to guide cellular responses and modulate NFkB pathway activation. The nano-tenosome, designed as a responsive system comprising exosomes and peptide, enables the investigation of the synergistic effects between the therapeutic molecules (cargo of the exosome) and magnetic forces. Thus, a synergic effect aims to control the dynamic release and target specific inflammatory pathway, thereby offering a strategic approach to controlling pathological inflammatory cues; iv)Validate the developed nano-tenosome system in preclinical tendinopathy models. The emphasis on combining different therapeutic cargos aims to address tendinopathy to promote healing and restore normal tendon function. Furthermore, the in vivo safety and biocompatibility tests will be performed for a correct validation of nano-tenosome. This approach aligns with the goal of demonstrating not only the regenerative potential but also the anti-inflammatory properties of the nano-tenosome.