From Single Forces to Integrated Signals: A Systematic Review of Endothelial Responses to Multifactorial Mechanics

ABSTRACT

The vascular endothelium is physiologically exposed to a concurrent field of mechanical stimuli, including shear stress (SS) resulting from blood flow, cyclic strain (CS) due to cardiac pulses, and substrate stiffness (ES) determined by the vessel wall. Despite this inherent complexity, in vitro mechanobiology research has predominantly relied on simplified, single‐stimulus paradigms. This systematic review, conducted in accordance with PRISMA guidelines, identifies a critical scarcity of multifactorial research on two or more concurrent mechanical constraints. Crucially, the data suggest that many experimental designs operate outside the functional in vivo bandwidth, with ranges contextualized as “physiological” often lacking consensus. Molecular mapping further demonstrates that combined forces frequently reverse or amplify the phenotypic effects of isolated cues. Furthermore, we identify critical methodological gaps, such as the over‐reliance on fetal venous cell models and the neglect of substrate viscoelasticity. Consequently, this work provides a novel mapping of integrated endothelial mechanotransduction and offers an actionable framework for improved experimental standardization. These findings have direct implications for the translational relevance of in vitro data in tissue engineering and vascular organoid development, emphasizing the necessity for multifactorial platforms that more faithfully recapitulate the nonlinear mechanical environment of the vasculature.