In the first stage of the project, I successfully developed an ultrafast and facile microfluidic technique to fabricate degradable synthetic microgels for cell encapsulation. The use of microfluidic technology enabled precise control over the size, uniformity, and architecture of the microgels, ensuring high reproducibility and scalability. By employing ultraviolet (UV)-induced photopolymerization, I achieved controlled crosslinking of the polymeric matrix, generating microgels with tunable mechanical stiffness and degradation kinetics. This degradation was modulated through the incorporation of protease-sensitive crosslinkers, allowing the microgels to degrade selectively in response to specific enzymatic cues that mimic physiological remodeling processes. As a proof of concept, human mesenchymal stem cells (hMSCs) were encapsulated within these synthetic microgels. The cells displayed excellent viability, structural integrity, and metabolic activity over extended culture periods, demonstrating that the engineered microenvironment effectively supports cell survival and function. These results validate the potential of the developed microgels as a fully synthetic, Matrigel-free platform for diverse regenerative medicine applications, including cell delivery, tissue repair, and organoid culture. These results were published in Advanced Healthcare Materials in 2023.
During the second phase of the project, I focused on optimizing the encapsulation of intestinal mesenchymal stem cells (IMCs) derived from human intestinal organoids within the degradable synthetic microgels. The encapsulated IMCs exhibited distinct secretory profiles depending on the mechanical properties of the microgels, revealing the ability of the synthetic platform to dynamically modulate cell behavior under both naïve and inflammatory conditions. Specifically, altering the macromer concentration in the polymeric network effectively tuned the cellular secretome, demonstrating the capacity of the matrix to guide regenerative and immunomodulatory responses. Finally, the regenerative capacity of the encapsulated IMCs was evaluated using advanced in vitro intestinal models, including human intestinal organoids. The cellular and molecular responses were analyzed through bulk RNA sequencing, which provided comprehensive insights into the transcriptional programs associated with tissue repair, extracellular matrix remodeling, and cytokine signaling. These findings collectively highlight the promise of synthetic degradable microgels as adaptive biomimetic matrices capable of supporting cell-driven regeneration in inflammatory environments. This project thus lays the foundation for the development of a novel cell-based regenerative therapy for IBD. By combining synthetic biomaterial engineering, stem cell biology, and organoid technology, the platform offers a clinically translatable strategy to restore intestinal integrity, enhance mucosal healing, and overcome the limitations of current anti-inflammatory treatments. In the long term, this work opens the door to the personalization of regenerative therapies, where patient-derived cells could be encapsulated within tailored synthetic microenvironments to promote durable tissue regeneration and improve patient outcomes in chronic intestinal disorders.
The results of this Action have been disseminated in 9 scientific conferences, several departamental seminars, one Open Access scientific publication, with other manuscript in preparation.
