ELECTRIFY: um adesivo multifuncional piezoelétrico para o tratamento da insuficiência cardíaca.

The overarching aim of ELECTRIFY is to develop and validate a therapeutic solution for advanced HF patients by associating an electroactive vascularized cardiac patch with drug-loaded nanoparticles (NPs) to promote vascular, electrical and contractile recovery beyond state-of-the-art EHT, while limiting local inflammation and fibrosis (Fig.2). Specific objectives (SO) are: SO1: Develop a 3D vascularized and electroactive cardiac patch. Vascularization units (VUs) will be combined with fibrin gel and used as building blocks to form a vascular network integrated in CM cultures established on piezoelectric nanofibers. The use of piezoelectric materials for heart repair is unique. Piezoelectric nanofibers will enhance CM maturation, alignment and electric stimulation [12] and, after implantation, will electrically stimulate the host myocardium, while VUs will support timely inosculation with the host, as previously reported [13]. The generated multicellular EHT will be also relevant for drug-screening and disease modelling and will be herein characterized as a fibrosis model. Key performance indicators (KPI): 1.1: CM contraction 15% higher on piezoelectric fibers; 1.2: CM orientation increased by > 75%; 1.3: VUs form a vascular network in the angiogenic assay; 1.4: CM contractility is not affected by VUs. SO2: Develop a NP-based anti-inflammatory/anti-fibrotic delivery system for the heart. Some anti-fibrotic drugs that have been tested in pre-clinical tests with relative success such as salinomycin [14] and pirfenidone [15]. Pirfenidone reduces fibrosis after MI by inhibiting fibroblast activation, inflammation and oxidative stress, constituting a great therapeutic candidate [16] and will, therefore, constitute our primary target. We will generate piezoelectric NPs (PIEZO NPs) that will deliver pirfenidone directly to the myocardium, avoiding reported gastrointestinal and liver side effects and improving bioavailability. This will be attained by capitalizing on PIEZO NPs deformation as a result of cardiac movements to trigger in-situ drug release. Local delivery of pirfenidone in the myocardium is unprecedented and will constitute a step towards precision in fibrosis control. KPI: 2.1: NPs will reduce (>20%) cardiac fibroblast activation in vitro without interfering with cell viability; 2.2: NPs reduce by 20% patch-surrounding myofibroblasts. SO3: Generate a multifunctional hybrid cardiac patch with anti-inflammatory/anti-fibrotic properties. Implanted EHT often become separated from the myocardium by scar tissue affecting their therapeutic efficacy. To overcome this, we will combine the two strategies described above to generate a cardiac patch able to modulate the adjacent environment towards less fibrosis. Pirfenidone delivery in conjugation with the cardiac patch will result in synergic benefits, namely reducing myocardial fibrosis and improving cell survival and overall construct integration. This combinatory approach is ground-breaking and, apart from advancing cardiac therapeutic development, we anticipate a broad impact on regenerative medicine and biomaterials, as inflammation/fibrosis is transversal to many organs. KPI: 3.1: Patch-treated mice recover from surgery (survival = to control group); 3.2: Decrease in left ventricle end-systolic volume (LVESV) by 10%; 3.3: 20% reduction on inflammatory and fibrotic cells surrounding the patch