Microfluidic Shear Processing Control of Biological Reduction Stimuli-Responsive Polymer Nanoparticles for Drug Delivery.

We demonstrate microfluidic manufacturing of glutathione (GSH)-responsive polymer nanoparticles (PNPs) with controlled in vitro pharmacological properties for selective drug delivery. This work leverages previous fundamental work on microfluidic control of the physicochemical properties of GSH-responsive PNPs containing cleavable disulfide groups in two different locations (core and interface, DualM PNPs). In this paper, we employ a two-phase gas-liquid microfluidic reactor for the flow-directed manufacturing of paclitaxel-loaded or DiI-loaded DualM PNPs (PAX-PNPs or DiI-PNPs, where DiI is a fluorescent drug surrogate dye). We find that both PAX-PNPs and DiI-PNPs exhibit similar flow-tunable sizes, morphologies, and internal structures to those previously described for empty DualM PNPs. Fluorescent imaging of DiI-PNP formulations shows that microfluidic manufacturing greatly improves the homogeneity of drug dispersion within the PNP population compared to standard bulk microprecipitation. Encapsulation of PAX in DualM PNPs significantly increases its selectivity to cancerous cells, with various PAX-PNP formulations showing higher cytotoxicity against cancerous MCF-7 cells than against non-cancerous HaCaT cells, in contrast to free PAX, which showed similar cytotoxicity in the two cell lines. In addition, the characterization of DualM PNP formulations formed at various microfluidic flow rates reveals that critical figures of merit for drug delivery function-including encapsulation efficiencies, GSH-triggered release rates, rates of cell uptake, cytotoxicities, and selectivity to cancerous cells-exhibit microfluidic flow tunability that mirrors trends in PNP size. These results highlight the potential of two-phase microfluidic manufacturing for controlling both structure and drug delivery function of biological stimuli-responsive nanomedicines toward improved therapeutic outcomes.

Controlled Microfluidic Synthesis of Biological Stimuli-Responsive Polymer Nanoparticles.

Microfluidic flow-directed self-assembly of biological stimuli-responsive block copolymers is demonstrated with dual-location cleavable linkages at the junction between hydrophilic and hydrophobic blocks and on pendant group within the hydrophobic blocks. On-chip self-assembly within a two-phase microfluidic reactor forms various "DualM" polymer nanoparticles (PNPs), including cylinders and multicompartment vesicles, with sizes and morphologies that are tunable with manufacturing flow rate. Complex kinetically trapped intermediates between shear-dependent states provide the most detailed mechanism to date of microfluidic PNP formation in the presence of flow-variable high shear. Glutathione (GSH)-triggered changes in PNP size and internal structure depend strongly on the initial flow-directed size and internal structure. Upon incubation in GSH, flow-directed PNPs with smaller average sizes showed a faster hydrodynamic size increase (attributed to junction cleavage) and those with higher excess Gibbs free energy showed faster inner compartment growth (attributed to pendant cleavage). These results demonstrate that the combination of chemical control of the location of biologically responsive linkages with microfluidic shear processing offers promising routes for tunable "smart" polymeric nanomedicines.