Carbon nanosheets (CNS) have emerged as a cutting-edge class of 2D nanostructured materials, boasting exceptional thermal and electrical conductivity alongside unique flake-like structures and reactive edge planes. Synthesized through diverse methods like solvothermal techniques, chemical vapor deposition (CVD), and chemical exfoliation, CNS exhibit high surface-to-volume ratios, ensuring numerous interaction points and making them highly desirable electrode materials. Of particular interest are nitrogen-doped (N-doped) CNS, which offer heightened electrocatalytic activity and biocompatibility, attributes crucial for biomedical applications. Nitrogen doping influences the physicochemical properties of carbonaceous materials, enhancing conductivity, catalytic activity, and altering crystal and electronic structures. However, challenges persist in achieving high nitrogen content and graphitization degrees without compromising the integrity of the nanostructures, owing to harsh synthesis conditions that can lead to the elimination of N heteroatoms and collapse of carbon nanostructures. Despite these hurdles, N-doped carbon nanosheets hold immense promise in biomedicine, especially in neurotransmitter-sensing applications. Their exceptional electrocatalytic activity, coupled with enhanced electrochemical stability and rapid ion transfer, positions them as prime candidates for neurotransmitter sensing in biomedical contexts. By leveraging these attributes, N-doped carbon nanosheets offer a pathway to significant advancements in diagnostic and therapeutic approaches, facilitating real-time monitoring and precise modulation of neurotransmitter levels in the central and peripheral nervous systems. Through ongoing research and refinement, the integration of N-doped carbon nanosheets into biomedical devices holds the potential to revolutionize the understanding and treatment of neurological disorders, opening new frontiers in personalized medicine and neuroengineering.