Toward Dynamic Biomimetic Biomaterials: Temporal Coupling of Flavonoid-Induced Cellular Responses and 3D-Printed PLA Scaffold Evolution.

Background/Objectives: Biomimetic biomaterials should be evaluated not only through their initial properties but also through their interaction with biological environments over time. This study investigated a human cell-based approach integrating flavonoid-induced cellular responses with the temporal evolution of 3D-printed polylactic acid (PLA) scaffolds under physiological-like conditions. Methods: Human Wharton's Jelly mesenchymal stem cells (WJ-MSCs) were exposed to naringin and hesperidin (250 µg/mL) and evaluated after 3, 7, and 10 days using ALP, TP, OPN, and OC. In parallel, PLA scaffolds' biodegradation was studied under static and dynamic conditions. Results: Naringin produced a more pronounced early/intermediate osteogenic-related response, whereas hesperidin showed comparatively more sustained ALP- and OPN-related activity at later stages. Dynamic scaffolds exhibited progressive temporal changes, with weight variations ranging from -0.384% to +0.127% at days 1, 3, 7, and 10, respectively, accompanied by progressive morphological modification. Initial AFM analysis showed an average RMS roughness of 22.1 nm. Conclusions: The distinct temporal profiles of the flavonoids, together with the evolving scaffold-medium interface, support a biomimetic strategy based on temporal coordination of biochemical cues. These findings provide a rationale for future experimentally validated sequential delivery systems designed to promote early osteogenic activation followed by sustained cellular and matrix-associated activity.