Mapping Nanoscale Protein-Corona Kinetics of DoE-Optimized Perfluorocarbon Encapsulated-PLGA Nanoparticles by In Situ, Time-Resolved Synchrotron SAXS.

Maria Joseph, Joice; Gigliobianco, Maria Rosa; Minnelli, Cristina; Moretti, Paolo; Spinozzi, Francesco; Mobbili, Giovanna; Pistolesi, Sara; Casadidio, Cristina et al. · Adv Healthc Mater · 2026

basic_science · Level V

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Abstract

Integrated frameworks that couple statistically guided formulation with nanoscale, time-resolved mapping of the protein corona (PC) remain rare for perfluoro-15-crown-5-ether-loaded poly(lactic-co-glycolic acid) nanoparticles (PFCE-PLGA NPs). We apply a two-phase workflow: a single-factor screen of PLGA/surfactant molecular-weight (Mw) pairings under homogenization versus probe sonication; and a central composite design varying sonication time, surfactant concentration, and PFCE volume to model effects on size, polydispersity index), zeta (ζ)-potential, loading capacity (LC), and fluorine signal-to-noise ratio. Scanning electron microscopy and transmission electron microscopy confirm spherical morphology of the NPs. Synchrotron small-angle X-ray scattering (SAXS) first resolves PFCE-dependent internal NP structure and then, in situ and time-resolved, tracks corona evolution across physiologic albumin levels (40, 20, and 2 mg mL<sup>-1</sup>), quantifying shell thickness, protein volume fraction, and polydispersity while distinguishing PFCE-PLGA from blank PLGA NPs. Fibroblast assays show no significant cytotoxicity at tested doses. Overall, combining design-of-experiments with in situ SAXS links processing variables to nanoscale structure and protein-mediated transformations, guiding rational PFCE-PLGA NP design.

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