Mucoadhesive microspheres are an advanced drug delivery system designed to enhance the bioavailability and therapeutic efficacy of drugs by adhering to mucosal surfaces. These microspheres offer prolonged residence time, controlled drug release, and targeted delivery to specific mucosal sites, making them ideal for oral, nasal, ocular, and vaginal applications. This review aims to provide a comprehensive overview of the formulation strategies, characterization techniques, and therapeutic applications of mucoadhesive microspheres. A thorough literature search was conducted using databases such as PubMed, Scopus, and Web of Science. Articles focusing on the development, evaluation, and clinical applications of mucoadhesive microspheres were included. The review covers the selection of polymers, methods of preparation, and characterization techniques for assessing mucoadhesive properties, particle size, drug encapsulation efficiency, and in vitro/in vivo drug release profiles. The review identified various natural and synthetic polymers, including chitosan, alginate, and carbopol, as effective mucoadhesive agents. Preparation techniques such as emulsification, spray drying, and solvent evaporation were discussed, highlighting their impact on the physical and chemical properties of the microspheres. Characterization methods such as scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and mucoadhesion testing were examined. Studies demonstrated that mucoadhesive microspheres significantly enhance drug absorption and bioavailability by adhering to mucosal tissues and providing controlled drug release. Therapeutic applications span across multiple fields, including gastroenterology, pulmonology, and ophthalmology, with promising results in improving patient compliance and treatment outcomes. Mucoadhesive microspheres represent a promising drug delivery system with the potential to revolutionize the administration of pharmaceuticals. Their ability to adhere to mucosal surfaces, provide controlled and sustained drug release, and enhance bioavailability makes them suitable for various therapeutic applications. Future research should focus on optimizing formulation parameters, exploring novel mucoadhesive polymers, and conducting clinical trials to establish their efficacy and safety. The advancements in mucoadhesive microspheres hold significant promise for improving drug delivery and therapeutic outcomes across different medical fields.