Wound healing assay on a gravity-driven unidirectional perfusion microfluidic device
Cell migration plays a pivotal role in a multitude of physiological and pathological processes, including tissue development, wound healing, immune response, and cancer metastasis. While traditional two-dimensional (2D) cell culture assays, such as the wound healing (scratch) test, have provided valuable insights into cell migration mechanisms, they are unable to replicate the complex in vivo environment. This study introduces a gravity-driven, pump-free microfluidic device to simulate in vivo flow conditions and to investigate the effects of flow rate and flow direction on wound healing assays. The regulation of the flow rate was achieved through adjustments to the tilt angle of the rocking plate. To evaluate the fluid dynamics and cell migration behavior under different flow conditions, numerical simulations and experimental analyses were conducted. The results showed that cell migration and wound healing were markedly influenced by the direction of fluid flow and the magnitude of flow rate. Specifically, compared to the static group, the wound healing rate increased by 76%±6% when the flow direction formed a 45° angle with the wound scratch axis at a -flow rate of 3.0 mL/min, corresponding shear stress of 0.38 dyne/cm 2 . Furthermore, cells exhibited enhanced alignment and migration in the direction of the fluid flow, with the most pronounced effects observed at angles of 45° and 90° between the scratch pattern and the flow direction. Analysis of the cell numbers within the wound healing area and measurements of individual cell area revealed that while the flow conditions did not enhance cell movement, they promoted cell spreading, making the collective wound healing region more pronounced. This gravity-driven, pump-free microfluidic device provides a cost-effective and physiologically relevant platform for studying cell migration, with potential applications in cancer research, wound healing, and developmental biology.
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