Real-time monitoring of relaxation and contractility of smooth muscle cells on a novel biohybrid chip

Abstract

Cardiovascular diseases represent the most common cause of death in industrialized countries. In this context vascular smooth muscle cells (SMCs) are a major key player that is involved in pathological processes like hypertension and atherosclerosis. Therefore the pharmaceutical industry is intensively investigated in developing non-destructive and label-free monitoring techniques for a quantitative detection of SMC characteristics in the field of active pharmaceutical development as well as clinical diagnostics. Hence, we developed a novel multiwell interdigital electrode sensor-array in standardized ANSI 96-well layout. Through optimization of electrode geometry and material as well as passivation/adhesion-layer we obtained a novel biohybrid chip for the sensitive and quantitative detection of SMC contractility as well as relaxation via impedance spectroscopy. For the validation of our multiwell sensor-array we established a SMC culture model derived from primary cells that is switchable from a non-contractile pathological to a functional contractile phenotype. Using the reference compounds acetylcholine (ACh) and amlodipine, we could quantify SMC contraction by an impedance decrease to 40% while SMC relaxation was detectable by an impedance increase to 110%. More strikingly we could monitor aging of the isolated SMC which arose by an attenuated contractility over successive passaging. Demonstrating the performance of our self-developed multiwell sensor-array based impedance measurement setup we provide a suitable sensor-array coupled cell model to study the mechanisms that activated SMCs undergo in response to inflammatory mediators or vessel injury.