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Here we developed a novel scaffold that significantly improves cardiac tissue regeneration by incorporating curved PCL fibers. These fibers mimic the natural architecture of the extracellular matrix (ECM), providing optimal mechanical and biochemical cues for cell growth. Using a brush-assisted bioprinting technique, we precisely aligned these fibers within a collagen-based scaffold to create a more physiologically relevant microenvironment. Our results demonstrate that the curved fiber design promotes better cell alignment, and differentiation of H9C2 cardiomyocytes compared to straight-fiber or collagen-only scaffolds. The enhanced performance is attributed to improved mechanotransduction and activation of key growth signaling pathways. Briefly, we used PCL and extrusion-based printing process create the micro-sized fibers. Through carefully control of the printing parameters, we were able to obtain a curved morphology of the fibers. These fibers were collected and formulated into a collagen-based bioink. Using a brush casting bioprinting system, highly aligned bioconstructs were obtained. Three types of samples: (i) scaffolds with curved fibers, (ii) scaffolds with straight fibers, and (iii) collagen scaffolds without any fibers for comparison. The live/dead assay indicated high cell survival (~90%), indicating that the method was safe for cells. Additionally, phalloidin staining showed highly aligned organization of cells in the direction of the brush movement. To further elucidate these findings, we measured gene expressions involved in mechanical sensing and cell development. Cells grown on curved fiber scaffolds showed upregulation of these genes compared to the control.
These findings suggests that the curved fiber evokes efficient mechanotransduction and help guide cell organization Our new brush-assisted printing method successfully created these beneficial curved fiber structures. The scaffolds provide both structural support and biological signals that heart cells need to grow and function properly. This research advances the field of cardiac tissue engineering in several ways. These findings could lead to better treatments for heart disease and damage in the future, though more research is needed to develop clinical applications.