Abstract Current hydrogel strain sensors have never been integrated into dynamic organ‐on‐a‐chip (OOC) due to the lack of sensitivity in aqueous cell culture systems. To enhance sensing performance, a novel strain sensor is presented in which the MXene layer is coated on the bottom surface of a pre‐stretched anti‐swelling hydrogel substrate of di‐acrylated Pluronic F127 (F127‐DA) and chitosan (CS) for isolation from the cell culture on the top surface. The fabricated strain sensors display high sensitivity (gauge factor of 290.96), a wide sensing range (0–100%), and high repeatability. To demonstrate its application, alveolar epithelial cells are cultivated on the top surface of the hydrogel strain sensor forming alveolar barriers, and then integrated into dynamic lung‐on‐a‐chip (LOC) systems. This system can sensitively monitor normal physiological breathing, pathological inflammation stimulated by lipopolysaccharide (LPS), and alleviated inflammation through drug intervention.