Effect of Nanopore #3 scaffold on gene expression on fibroblasts

Scaffolds with pores can influence cellular fate and tissue microenvironments by altering intracellular and intercellular signaling pathways, which are crucial for guiding tissue regeneration. Despite their significance, the cellular mechanisms behind the response to pores, especially at the nanoscale, are not well understood. Our study shows that scaffolds with different nanopore characteristics can enhance skin regeneration in various ways, with fibroblasts being the primary cellular responders. We have demonstrated that caveolae formation is a critical mechanism by which fibroblasts interact with nanopores. The phosphorylation of caveolin-1 (CAV1) is a key event in this process, enabling caveolae-mediated endocytosis and the subsequent internalization of cellular substances. This initiates a cascade of events involving the internalization of cell surface receptors such as PDGFRβ, activation of tyrosine kinase pathways including PI3K, AKT, and ERK1/2, and the phosphorylation of the transcription factor RUNX1. The nuclear translocation of RUNX1 upregulates the expression of fibroblast growth factor binding protein 3 (FGFBP3), which is a key factor in tissue repair. Additionally, cytoplasmic CAV1 can competitively bind to Filamin-A, releasing CBFβ, which then translocates to the nucleus and enhances RUNX1's DNA-binding affinity, synergistically activating FGFBP3 transcription and promoting tissue healing. Collectively, our findings underscore the importance of considering nanopore characteristics in scaffold design for tissue regeneration and highlight caveolae formation as a central mediator of cellular responses to nanoporous environments, initiating a multitude of biological processes essential for tissue repair and regeneration.