To build foundational expertise in nanomedicine synthesis while accommodating the diverse research areas represented across projects, trainees select one of three specialization tracks: lipid-, polymer-, or metal-based nanoparticle systems. Each platform has distinct physicochemical properties and requires specific synthesis methods, protocols, and technologies. Accordingly, each track combines tailored theoretical instruction with hands-on laboratory training. Designed for trainees from diverse disciplinary backgrounds, including those with limited or no prior experience in chemistry, materials science, or nanoparticle synthesis, these modules provide the foundational knowledge and practical skills needed to understand and apply key principles of nanomedicine synthesis.

Like Module 1, Module 2 is designed to accommodate trainees from diverse disciplinary backgrounds, including those with limited or no prior experience in nanoparticle synthesis or the chemical sciences, thereby fostering interdisciplinary learning and collaboration. Training focuses on the physicochemical properties that govern nanoparticle stability and interactions with biological environments. Through hands-on training with state-of-the-art instrumentation in participating laboratories, trainees develop practical skills in nanoparticle characterization while gaining an understanding of key quality control principles, including lot-to-lot reproducibility and the quality of starting materials.

Module 2 covers a broad range of characterization and quality control methods, including sterilization mechanisms and technologies; dialysis, size-exclusion chromatography, and filtration; detection of microbial and endotoxin contamination; and measurement of osmolality and viscosity. Trainees also gain experience in sample preparation, instrument operation, and data analysis using techniques such as Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA), Transmission and Scanning Electron Microscopy (TEM/SEM), Single Particle Analysis (SPA), Mass Spectrometry (MS), zeta potential analysis, and spectroscopic and elemental analysis. Additional topics include the characterization and quantification of excipients, lipid and polymer components, counterions, nanoparticle surface coatings, drug and RNA content and distribution as a function of nanoparticle size, as well as ligand concentration and targeting efficiency.

Module 3 focuses on nanoparticle interactions with biological environments and the integration of emerging technologies into analytical workflows. Trainees develop a theoretical understanding of the chemical, biochemical, and molecular composition of biological fluids, including blood plasma, lymph, vitreous fluid, urine, and synovial fluid, and how these environments influence nanomedicine stability, performance, and biological interactions.

Training covers key principles for evaluating nanoparticle stability under biologically relevant and storage conditions, including changes in particle size and aggregation over time; leakage and degradation of active pharmaceutical ingredients, such as drugs and RNA; degradation of lipid-, polymer-, and metal-based nanoparticles; RNA integrity and transferability; and drug or prodrug release in plasma. Trainees also examine the effects of solvent conditions, temperature, pH, light exposure, freeze-thaw cycles, lyophilization, centrifugation, and filtration, as well as approaches for establishing appropriate storage conditions and shelf life.

Module 3 further introduces trainees to the biological evaluation of nanomedicines through cytotoxicity and cell viability testing; inflammation and immunogenicity assessment; and biofluid-, cell-, and tissue-based assays. Additional topics include routes of nanoparticle administration and their physiological rationale; the fundamentals of optical and nuclear biomedical imaging for assessing nanoparticle distribution; and the evaluation of nanoparticle pharmacokinetics, clearance, and biodistribution in vivo using preclinical small-animal models.

Together, these components provide trainees with a comprehensive understanding of how nanoparticle properties, formulation conditions, and biological environments influence the stability, safety, and performance of nanomedicines.