Organ-on-chip (OOC) technology is an innovative approach that reproduces human organ structures and functions on microfluidic platforms, offering detailed insights into physiological processes. This technology provides unique advantages over conventional in vitro and in vivo models, making it a promising tool for biomedical research and drug screening. The review compares OOCs with conventional models, highlighting their differences and applications in biomedical research. OOCs offer more physiological complexity and experimental controllability compared to 2D/3D cell cultures and animal models, which lack the ability to fully replicate human physiology and pathology. OOCs can simulate drug delivery and penetration, regulate fluid movements, and provide real-time visualization and quantitative analysis of human biological processes. The review also discusses the development of multi-organ systems (MOCs) within OOC technology, which integrate multiple OOCs to study systemic interactions between organs. These systems offer more realistic and physiologically relevant in vitro models, but they face challenges such as the development of blood mimetic media, standardization of manufacturing processes, and achieving physiologically relevant conditions. Despite these challenges, OOC technology is rapidly evolving and aligns with changing regulatory paradigms, offering potential for personalized medicine and advanced drug discovery. The future of OOC technology holds promise for reshaping biomedical research and improving drug efficacy assessments.Organ-on-chip (OOC) technology is an innovative approach that reproduces human organ structures and functions on microfluidic platforms, offering detailed insights into physiological processes. This technology provides unique advantages over conventional in vitro and in vivo models, making it a promising tool for biomedical research and drug screening. The review compares OOCs with conventional models, highlighting their differences and applications in biomedical research. OOCs offer more physiological complexity and experimental controllability compared to 2D/3D cell cultures and animal models, which lack the ability to fully replicate human physiology and pathology. OOCs can simulate drug delivery and penetration, regulate fluid movements, and provide real-time visualization and quantitative analysis of human biological processes. The review also discusses the development of multi-organ systems (MOCs) within OOC technology, which integrate multiple OOCs to study systemic interactions between organs. These systems offer more realistic and physiologically relevant in vitro models, but they face challenges such as the development of blood mimetic media, standardization of manufacturing processes, and achieving physiologically relevant conditions. Despite these challenges, OOC technology is rapidly evolving and aligns with changing regulatory paradigms, offering potential for personalized medicine and advanced drug discovery. The future of OOC technology holds promise for reshaping biomedical research and improving drug efficacy assessments.