Quantifying nanoparticle delivery: challenges, tools, and advances.

This review explores challenges and methods for quantifying nanoparticle delivery in therapeutic applications. We discuss three main approaches: (1) functional readouts that assess therapeutic effects post nanoparticle administration, (2) nanocarrier tracking that directly monitors the nanoparticle localization, and (3) cargo tracking that infers nanoparticle localization by measuring encapsulated agents or attached surface tags. Reanalysis of the Wilhelm et al. Cancer Nanomedicine Repository dataset found mixed quantification methodologies, which could cause misleading conclusions. We discuss potential pitfalls in each quantification approach and highlight recent advancements in novel technologies. It is important that researchers select appropriate quantification methods based on their objectives and consider integrating multiple approaches for a comprehensive understanding of in vivo nanoparticle behavior to facilitate their clinical translation.