Multifunctional Nanovesicle-Bioactive Conjugates Prepared by a One-Step Scalable Method Using CO₂-Expanded Solvents

Abstract

The integration of therapeutic biomolecules, such as proteins and peptides, in nanovesicles is a widely used strategy to improve their stability and efficacy. However, the translation of these promising nanotherapeutics to clinical tests is still challenged by the complexity involved in the preparation of functional nanovesicles and their reproducibility, scalability, and cost production. Here we introduce a simple one-step methodology based on the use of CO2-expanded solvents to prepare multifunctional nanovesicle-bioactive conjugates. We demonstrate high vesicle-to-vesicle homogeneity in terms of size and lamellarity, batch-to-batch consistency, and reproducibility upon scaling-up. Importantly, the procedure is readily amenable to the integration/encapsulation of multiple components into the nanovesicles in a single step and yields sufficient quantities for clinical research. The simplicity, reproducibility, and scalability render this one-step fabrication process ideal for the rapid and low-cost translation of nanomedicine candidates from the bench to the clinic.

Benznidazole Nanoformulates: A Chance to Improve Therapeutics for Chagas Disease

“…33 Ethanol and nonencapsulated BNZ were removed by diafiltration of nanovesicular suspensions. CDs loaded with BNZ were prepared by an antisolvent precipitation with compressed CO 2 .…”

Nanostructuring Lipophilic Dyes in Water Using Stable Vesicles, Quatsomes, as Scaffolds and Their Use as Probes for Bioimaging

“…Dye‐loaded QSs can be prepared by a one‐step method using compressed CO 2 , named depressurization of expanded liquid organic solution‐suspension (DELOS‐SUSP), showing several advantages over conventional routes for the preparation of functionalized vesicles. Indeed, it is a green technology leading in a single step to the formation of multifunctional nanovesicles with superior structural homogeneity . As dye models, we used lipophilic fluorophores with long alkyl chains, which can be integrated into QS membrane exploiting hydrophobic interactions between the chains of the dyes and the hydrophobic compartment of the double‐layer membrane.…”

“…DiD‐loaded QSs with DID loadings, L = moles DID /(moles CTAB + moles Cholesterol ), ranging from 0.6 × 10 −3 to 6.6 × 10 −3 (Table S1, Supporting Information) were prepared using the DELOS‐SUSP methodology . The physicochemical properties of the resulting vesicles with L = 1.3 × 10 −3 (D‐QS‐2) are here compared with those of other DiD‐loaded QSs obtained by other methods generally used for the preparation of loaded vesicles, such as incubation (D‐QS‐2‐IC), sonication (D‐QS‐2‐SON), and thin‐film hydration (D‐QS‐2‐TFH) .…”

“…These measurements show that the size, colloidal stability, and photophysical properties of dye‐loaded QSs make them remarkable candidates as nanostructured probes for biological imaging. These results together with the capability of QSs to integrate/encapsulate small drugs or large biomolecules and to decorate their surfaces with targeting groups open the possibility of producing in the near‐future multifunctional vesicles for theranostic applications.…”

Polymer Capsules as a Theranostic Tool for a Universal In Vitro Screening Assay-The Case of Lysosomal Storage Diseases

“…Recently, nanoliposomes loaded with GLA were introduced to effectively deliver the recombinant protein to primary Fabry cells. It was demonstrated that the carried enzymes exhibited a higher specific activity and better enzymatic performance than free administered enzymes …”