Microfluidic-Generated Immunomodulatory Nanoparticles and Formulation-Dependent Effects on Lipopolysaccharide-Induced Macrophage Inflammation

Abstract

Nanoparticles (NPs) have emerged as a highly useful and clinically translatable drug delivery platform for vast therapeutic payloads. Through the precise tuning of their physicochemical properties, NPs can be engineered to exhibit controlled drug release properties, enhanced circulation times, improved cellular uptake and targeting, and reduced toxicity profiles. Conventional bulk methods for the production of polymeric NPs suffer from the ability to control their size and polydispersity, batch-to-batch variability, significant preparation times, and low recovery. Here, we describe the development and optimization of a high-throughput microfluidic method to produce cargo-less immunomodulatory nanoparticles (iNPs) and their formulation-dependent anti-inflammatory properties for the modulation of lipopolysaccharide (LPS)-induced macrophage responses. Using poly(lactic acid) (PLA) as the core-forming polymer, a rapid and tunable microfluidic hydrodynamic flow-focusing method was developed and optimized to systematically evaluate the role of polymer and surfactant concentration, surfactant chemistry, and flow rate ratio (FRR) on the formation of iNPs. A set of iNPs with 6 different surface chemistries and 2 FRRs was then prepared to evaluate their inherent anti-inflammatory effects using bone marrow-derived macrophages stimulated with the Toll-like receptor 4 agonist, LPS. Finally, a lyophilization study was performed using various cryoprotectants and combinations to identify preferable conditions for iNP storage. Overall, we demonstrate a highly controlled and reproducible method for the formulation of iNPs using microfluidics and their formulation-dependent inherent anti-inflammatory immunomodulatory properties, which represents a potentially promising strategy for the management of inflammation.

Nanoparticle dose and antigen loading attenuate antigen‐specific T‐cell responses

“…PLG nanoparticles are being investigated as alternative Ag‐carriers that are biodegradable, tunable, and manufacturable at large scale (Pearson, Casey, Hughes, Miller, et al, 2017), (Truong et al, 2021). Ag has been incorporated into NPs by encapsulation, surface conjugation, and as Ag‐polymer conjugates (D. R. Getts et al, 2012; McCarthy et al, 2017; Pearson, Casey, Hughes, Miller, et al, 2017; Tostanoski et al, 2016).…”

Lipid–Polymer Hybrid Nanoparticles Utilize B Cells and Dendritic Cells to Elicit Distinct Antigen-Specific CD4+ and CD8+ T Cell Responses

“…Rational biomaterial selection can also have inherent immune-stimulating or immune-modulating effects. For example, a palmitoyl-based lipid coupled to a peptide Ag can directly upregulate immunity. , PLGA particles have demonstrated the intrinsic ability to prevent DC and macrophage maturation and block immune-stimulating signaling pathways. − Rationally conjugating Ags to either lipids or polymers and encapsulating in NPs can provide a versatile tool to expand delivery mechanisms and control desired T cell immunity or tolerance.…”

Nanocarriers for effective delivery: modulation of innate immunity for the management of infections and the associated complications

“…Chavez-Santoscoy et al developed a functionalized polyanhydride nanoparticles and found that the nanoparticles initiated the secretion of pro-inflammatory cytokines such as IL-6, IL-1b and TNF-α in alveolar macrophages [260]. Recently, Truong et al developed a high-throughput microfluidic method for the generation of cargo-less immunomodulatory polymeric nanoparticles [261]. In this study, a set of 12 nanoparticles were developed and their ability to modulate the LPS-induced macrophage responses were investigated.…”