In Silico Evaluation of Mechanobiological Parameters Under Variable Flow in Three-Dimensional Microfluidic Platforms...

Background: Cell migration is a biological process influenced by biochemical signals and mechanical stimuli from the microenvironment. In this context, the accurate characterization of the mechanical microenvironment generated within microfluidic platforms represents an essential step for the design and interpretation of cell migration studies. Understanding how hydrodynamic forces influence the mechanical microenvironment experienced by cells remains a challenge, especially in confined and biomimetic systems. Methods: In this study, a three-dimensional microfluidic device was developed in silico to characterize the effects of flow variation on mechanofluidic parameters and to provide a quantitative basis for designing future cell-migration experiments. Computational fluid dynamics simulations were performed to characterize the velocity, pressure, and wall shear stress (WSS) distributions under different inlet flow rates (0.5, 1, and 5 µL/min) and three distinct inlet/outlet configurations within the same three-dimensional geometry. Rigid hemispherical probe structures were incorporated into the model to quantify the local shear stress acting on cell-sized surfaces. Results: The results demonstrated a direct and linear relationship between the applied flow rate and the WSS, modulated by the channel geometry and the inlet and outlet configuration. Regions near micropores and lateral channels showed high WSS values, while central regions experienced less mechanical stimulation, depending on flow conditions. Comparison with WSS values and ranges associated with cellular responses reported in the literature indicated that certain operational configurations generated mechanical conditions comparable to those previously investigated in cell-based studies, including cell migration applications. Conclusions: Overall, the study highlights the importance of controlling flow conditions in microfluidic platforms and provides a quantitative basis for the development and optimization of three-dimensional microfluidic devices intended for designing future cell-migration experiments. The systematic comparison of three inlet/outlet configurations across three flow rates within the same three-dimensional geometry provides a comparative framework for identifying configuration-dependent changes in the local mechanofluidic environment, supporting the selection of operational conditions for future mechanobiological and cell-migration studies.