title: Three-dimensional hydrodynamic focusing method for polyplex synthesis.
abstract: Successful intracellular delivery of nucleic acid therapeutics relies on multiaspect optimization, one of which is formulation. While there has been ample innovation on chemical design of polymeric gene carriers, the same cannot be said for physical processing of polymer-DNA nanocomplexes (polyplexes). Conventional synthesis of polyplexes by bulk mixing depends on the operators' experience. The poorly controlled bulk mixing process may also lead to batch-to-batch variation and consequent irreproducibility. Here, we synthesize polyplexes by using a three-dimensional hydrodynamic focusing (3D-HF) technique in a single-layered, planar microfluidic device. Without any additional chemical treatment or postprocessing, the polyplexes prepared by the 3D-HF method show smaller size, slower aggregation rate, and higher transfection efficiency, while exhibiting reduced cytotoxicity compared to the ones synthesized by conventional bulk mixing. In addition, by introducing external acoustic perturbation, mixing can be further enhanced, leading to even smaller nanocomplexes. The 3D-HF method provides a simple and reproducible process for synthesizing high-quality polyplexes, addressing a critical barrier in the eventual translation of nucleic acid therapeutics.
authors.affiliation: Department of Engineering Science and Mechanics, The Pennsylvania State University , University Park, Pennsylvania 16802, United States.
authors.forename: Mengqian
authors.initials: M
authors.lastname: Lu
authors.forename: Yi-Ping
authors.initials: YP
authors.lastname: Ho
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Grigsby
authors.forename: Ahmad Ahsan
authors.initials: AA
authors.lastname: Nawaz
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
authors.forename: Tony Jun
authors.initials: TJ
authors.lastname: Huang
mesh_terms: D000162:Acoustics; D057446:Hydrodynamics; D007700:Kinetics; D011108:Polymers / Q000737:chemistry*; D013696:Temperature
chemical_list: D011108:Polymers