The organ-on-a-chip (OoC) technology holds significant promise for biosensors and personalized medicine by enabling the creation of miniature, patient-specific models of human organs. This review studies the recent advancements in the application of polydimethylsiloxane (PDMS) microfluidics for OoC purposes. It underscores the main fabrication technologies of PDMS microfluidic systems, such as photolithography, injection molding, hot embossing, and 3D printing. The review also highlights the crucial role of integrated biosensors within OoC platforms. These electrochemical, electrical, and optical sensors, integrated within the microfluidic environment, provide valuable insights into cellular behavior and drug response. Furthermore, the review explores the exciting potential of PDMS-based OoC technology for personalized medicine. OoC devices can forecast drug effectiveness and tailor therapeutic strategies for patients by incorporating patient-derived cells and replicating individual physiological variations, helping the healing process and accelerating recovery. This personalized approach can revolutionize healthcare by offering more precise and efficient treatment options. Understanding OoC fabrication and its applications in biosensors and personalized medicine can play a pivotal role in future implementations of multifunctional OoC biosensors.