Project Portugal 2030

Summary

Hypothesis: Pathological calcification occurring in tissues is a common disorder that leads to pain and decreased quality of life. Discussions with shoulder clinician and surgeons attested that the state-of-the art treatment solutions for severe calcific pathology are in many cases not adequate to face this debilitating and painful tendinopathic disease. This fact motivates the development of early therapeutic interventions in tissue degeneration, in order to prevent the development of abnormal differentiations (such as osteogenesis/calcified deposits) and promote normal tissue healing. A key element is the aberrant growth of mineralization in the new tissue and, by modeling these events, we can better understand the interplay between healthy/pathological growth/development in tendons. How does modulation of mechanotransduction affect the prevention of calcification? The study of the mechanisms regulating tendon differentiation continues to be a critical and necessary issue in the literature, in order to develop efficient therapies. MagSphere raises the hypothesis that magnetically activating the TGF-ß/Smad2/3 cascade and antagonizing the BMP pathway represents an effective strategy to control/promote tenogenic homeostasis, this way preventing abnormal ossification, which plays an important role in the pathogenesis of tissue calcification. Few engineered models reflecting the interactive nature of signaling molecules in tendon have so far been addressed. MagSphere aim at develop bioengineered tendon spheroids that will be integrated in a circulatory perfusion system, facilitating dynamic responses and relevant readouts. Objectives: To develop an in vitro calcification model that allow the re-creation of healthy and pathological tissue, in order to allow the study of calcification/mineralization mechanisms, extra and intracellular mechanisms, as well as native multicellular crosstalk. This model can also serve as screening platform for the development of advanced therapies, mimicking cellular, biomolecular, biochemical and biophysical characteristics of tissue pathophysiology. MagSphere will enable a deeper understanding of molecular and cell mechanisms necessary to obtain successful tendon oriented therapies. The project explores novel concepts and goes across different disciplines, using one of the top emerging technologies that is “organ-on-chip” technology. The creation of 3D spheroids bridged with on-chip microbioreactors offers the possibility to work with complex cell cultures while providing better engineered microenvironments, maximizing the health/disease model. O1: Construction of magnetic spheroids with adequate cell sources, and characterization O2: Fabricate perfusable microbioreactor to culture/study magnetic spheroids on chip O3: Investigate the in vitro calcification model that allow the re-creation of healthy or pathological tissue O4: Study the modulation of TGF-B signaling cascades to trigger calcification resolution responses Expected outcomes: Insights at modulating calcification and inflammation towards proper healing present a breakthrough in tendinopathy treatments resulting in significant healthcare, economical and societal impact. We anticipate i) the identification of signaling targetable components that counter calcific tendinopathy or even boost tendon regeneration, as well as ii) creation of theranostics platform at personalized medicine strategies, customizing both diagnosis and treatment profiles.