Engineering 3D in vitro tissue models through integrated computational and experimental approaches

This PhD research focuses on the development, modeling, and optimization of advanced three-dimensional (3D) in vitro models designed to replicate complex tissue microenvironments, with particular attention to breast cancer. By integrating experimental and computational approaches, the project aims to bridge the gap between in vitro and in silico methodologies, enhancing the predictive power of preclinical models while minimizing the reliance on animal testing. Initially, computational fluid dynamics (CFD) simulations were applied to analyze nutrient diffusion within porous scaffolds supporting sensitive neuronal and glial cells. These studies emphasized the role of microstructural design in ensuring cell viability under static conditions. Building on these insights, the core of the research focused on engineering compartmentalized core-shell hydrogels made of alginate, designed to mimic the stiffness and biochemical gradients of the tumor microenvironment (TME). These constructs demonstrated biologically relevant features, including oxygen gradients and hypoxia markers, and were validated using human breast cancer cells. The incorporation of the engineered hydrogels into organ-on-chip systems enabled dynamic perfusion, which significantly improved nutrient and drug distribution, and more accurately replicated in vivo-like drug responses. In particular, cisplatin treatment under flow conditions revealed more homogeneous cytotoxic effects compared to static cultures. Parallel CFD analyses provided predictive insights into drug and nutrient transport, further optimizing culture conditions and model design. This work demonstrates that the integration of biomimetic hydrogels, fluid-dynamic stimulation, and computational modeling represents a robust strategy for the development of physiologically relevant in vitro models. The resulting platform improves therapeutic screening reliability and supports regulatory shifts towards reducing animal use in drug testing. Overall, the research provides a versatile and ethically responsible tool for advancing tissue engineering and cancer research.