Project Portugal 2030

Summary

The SOFT-CardioDoC specific objectives (SO) are: SO1. Selection and characterization of hydrogel/bioinks: Ensuring optimal performance of the enzymatic-crosslinked silk-fibroin hydrogel-based microfluidic platforms and formulations comprising elastin, gelatin methacrylated, and collagen may require extensive optimization of fabrication techniques, material properties, and experimental protocols. In particular, fine-tuning the mechanical properties of the enzymatic-crosslinked hydrogels to mimic the dynamic nature of the cardiac 3D microenvironment. We will investigate their structure, mechanical properties, injectability, swelling behavior, gelation time, and biocompatibility. SO2. Development of Soft and Vascularized Microfluidics Platforms: Design and fabrication of soft microfluidic platforms using proprietary silk fibroin enzymatic-crosslinked hydrogels to overcome limitations associated with traditional microfluidics. Set-up the microfabrication methods such as stereolitography and digital light processing 3D printing for development of an intricate vascularized microfluidic perfusable and transparent platform essential to demonstrate the reliability, reproducibility, and relevance of the engineered cardiac tissues. Achieve an adequate structural stability to prevent device deformation and ensure reproducibility of experimental results over time and allow integration with microscopic techniques. Investigate protein adsorption by enzyme-linked immunosorbent assay and fluorescent labeling. SO3. Modeling Cardiac Microenvironment: Incorporation of biological entities such as stem cells, primary cardiomyocytes, endothelial cells, fibroblasts and extracellular matrix components, into the soft microfluidic platforms to create biomimetic artificial cardiac tissues. Optimization of fluid perfusion and culture conditions to simulate physiological conditions and vascularized architectures. The engineered models will facilitate the study of 3D cardiac microenvironments, cardiac fibrosis, and arrhythmias, providing valuable insights into disease mechanisms and potential therapeutic interventions. We will use immunohistological techniques, dynamic mechanical analysis to observe changes in the microenvironment, production of extracellular matrix, and mechanical behavior of the engineered cardiac tissues during culturing and experimental testing. Cell-materials interactions will be investigated by examining cell behavior, phenotype, and function within the soft microfluidic environments through immunofluorescence staining, live-cell imaging, and functional assays. SO4. Drug Testing: The biomimetic cardiac tissues can serve as reliable platforms for drug testing, offering a more accurate representation of in vivo responses. The proof-of-concept study using the engineered cardiac in vitro models on a chip for drug testing applications will be carried out, offering a more physiologically relevant platform for investigating heart diseases, arrhythmias and the screening of potential therapeutic interventions in a controlled laboratory setting. We will use gene expression profiling, contractility measurements and electrical activity of cardiomyocytes, enabling the detection of drug-induced alterations in cardiac tissues offering insights into their potential negative impact on both cardiac performance and electrophysiology, and identify potential cardiotoxicity risks based on drug properties, and pharmacokinetics/pharmacodynamics.