Nanoparticle-Protein Interaction: Demystifying the Correlation between Protein Corona and Aggregation Phenomena.

Protein corona formation and nanoparticles' aggregation have been heavily discussed over the past years since the lack of fine-mapping of these two combined effects has hindered the targeted delivery evolution and the personalized nanomedicine development. We present a multitechnique approach that combines dynamic light and small-angle X-ray scattering techniques with cryotransmission electron microscopy in a given fashion that efficiently distinguishes protein corona from aggregates formation. This methodology was tested using ∼25 nm model silica nanoparticles incubated with either model proteins or biologically relevant proteomes (such as fetal bovine serum and human plasma) in low and high ionic strength buffers to precisely tune particle-to-protein interactions. In this work, we were able to differentiate protein corona, small aggregates formation, and massive aggregation, as well as obtain fractal information on the aggregates reliably and straightforwardly. The strategy presented here can be expanded to other particle-to-protein mixtures and might be employed as a quality control platform for samples that undergo biological tests.

Protein corona meets freeze-drying: overcoming the challenges of colloidal stability, toxicity, and opsonin adsorption.

Freeze-drying of nanoparticle suspensions is capable of generating stable nanoformulations with improved storage times and easier transportation. Nonetheless, nanoparticle aggregation is likely induced during freeze-drying, which reduces its redispersibility upon reconstitution and leads to undesirable effects such as non-specific toxicity and impaired efficacy. In this work, bovine serum albumin (BSA) is described as a suitable protectant for silica nanoparticles (SNPs), which result in solid structures with excellent redispersibility and negligible signs of aggregation even when longer storage times are considered. We experimentally demonstrated that massive system aggregation can be prevented when a saturated BSA corona around the nanoparticle is formed before the lyophilization process. Furthermore, the BSA corona is able to suppress non-specific interactions between these nanoparticles and biological systems, as evidenced by the lack of residual cytotoxicity, hemolytic activity and opsonin adsorption. Hence, BSA can be seriously considered for industry as an additive for nanoparticle freeze-drying since it generates solid and redispersible nanoformulations with improved biocompatibility.

Colloidal Stability and Redispersibility of Mesoporous Silica Nanoparticles in Biological Media.

The outreach of nanoparticle-based medical treatments has been severely hampered due to the imbalance between the efforts in designing extremely complex materials and the general lack of studies devoted to understanding their colloidal stability in biological environments. Over the years, the scientific community has neglected the relevance related to the nanoparticles' colloidal state, which consequently resulted in very poor bench-to-clinic translation. In this work, we show how mesoporous silica nanoparticles (MSNs, one of the most promising and tested drug delivery platforms) can be efficiently synthesized and prepared, resulting in a colloidally stable system. We first compared three distinct methods of template removal of MSNs and evaluated their ultimate colloidal stability. Then, we also proposed a simple way to prevent aggregation during the drying step by adsorbing BSA onto MSNs. The surface modification resulted in colloidally stable particles that are successfully redispersed in biologically relevant medium while retaining high hemocompatibility and low cytotoxicity.