title: Scalable fabrication of size-controlled chitosan nanoparticles for oral delivery of insulin.
abstract: Controlled delivery of protein would find diverse therapeutic applications. Formulation of protein nanoparticles by polyelectrolyte complexation between the protein and a natural polymer such as chitosan (CS) is a popular approach. However, the current method of batch-mode mixing faces significant challenges in scaling up while maintaining size control, high uniformity, and high encapsulation efficiency. Here we report a new method, termed flash nanocomplexation (FNC), to fabricate insulin nanoparticles by infusing aqueous solutions of CS, tripolyphosphate (TPP), and insulin under rapid mixing condition (Re > 1600) in a multi-inlet vortex mixer. In comparison with the bulk-mixing method, the optimized FNC process produces CS/TPP/insulin nanoparticles with a smaller size (down to 45 nm) and narrower size distribution, higher encapsulation efficiency (up to 90%), and pH-dependent nanoparticle dissolution and insulin release. The CS/TPP/insulin nanoparticles can be lyophilized and reconstituted without loss of activity, and produced at a throughput of 5.1 g h-1when a flow rate of 50 mL min-1is used. Evaluated in a Type I diabetes rat model, the smaller nanoparticles (45 nm and 115 nm) control the blood glucose level through oral administration more effectively than the larger particles (240 nm). This efficient, reproducible and continuous FNC technique is amenable to scale-up in order to address the critical barrier of manufacturing for the translation of protein nanoparticles.
authors.affiliation: School of Materials Science and Engineering, and Center of Functional Biomaterials, Sun Yat-sen University, Guangzhou 510275, PR China; School of Medicine, Sun Yat-sen University, Guangzhou, 510080, PR China.
authors.forename: Zhiyu
authors.initials: Z
authors.lastname: He
authors.affiliation: Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Santos
authors.affiliation: School of Materials Science and Engineering, and Center of Functional Biomaterials, Sun Yat-sen University, Guangzhou 510275, PR China.
authors.forename: Houkuan
authors.initials: H
authors.lastname: Tian
authors.affiliation: School of Materials Science and Engineering, and Center of Functional Biomaterials, Sun Yat-sen University, Guangzhou 510275, PR China.
authors.forename: Huahua
authors.initials: H
authors.lastname: Huang
authors.affiliation: Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Yizong
authors.initials: Y
authors.lastname: Hu
authors.affiliation: School of Materials Science and Engineering, and Center of Functional Biomaterials, Sun Yat-sen University, Guangzhou 510275, PR China.
authors.forename: Lixin
authors.initials: L
authors.lastname: Liu
authors.affiliation: School of Materials Science and Engineering, and Center of Functional Biomaterials, Sun Yat-sen University, Guangzhou 510275, PR China; School of Medicine, Sun Yat-sen University, Guangzhou, 510080, PR China; Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA. Electronic address: kam.leong@columbia.edu.
authors.forename: Kam W
authors.initials: KW
authors.lastname: Leong
authors.affiliation: School of Materials Science and Engineering, and Center of Functional Biomaterials, Sun Yat-sen University, Guangzhou 510275, PR China; School of Chemistry, Key Laboratory for Polymeric Composite and Functional Materials of Ministry of Education, Sun Yat-sen University, Guangzhou 510275, PR China. Electronic address: chenym35@mail.sysu.edu.cn.
authors.forename: Yongming
authors.initials: Y
authors.lastname: Chen
authors.affiliation: School of Materials Science and Engineering, and Center of Functional Biomaterials, Sun Yat-sen University, Guangzhou 510275, PR China; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Translational Tissue Engineering Center, and Whitaker Biomedical Engineering Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. Electronic address: hmao@jhu.edu.
authors.forename: Hai-Quan
authors.initials: HQ
authors.lastname: Mao
mesh_terms: D000284:Administration, Oral; D000818:Animals; D001692:Biological Transport; D001786:Blood Glucose / Q000378:metabolism; D018938:Caco-2 Cells; D048271:Chitosan / Q000737:chemistry*; D003921:Diabetes Mellitus, Experimental / Q000097:blood / Q000188:drug therapy / Q000473:pathology; D004195:Disease Models, Animal; D016503:Drug Delivery Systems; D017097:Electric Impedance; D005612:Freeze Drying; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000008:administration & dosage* / Q000493:pharmacokinetics / Q000627:therapeutic use; D008297:Male; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D010316:Particle Size; D011122:Polyphosphates / Q000737:chemistry; D017207:Rats, Sprague-Dawley; D034421:Sus scrofa; D019108:Tight Junctions / Q000378:metabolism
chemical_list: D001786:Blood Glucose; D007328:Insulin; D011122:Polyphosphates; D048271:Chitosan; C005692:triphosphoric acid
keywords: Chitosan nanoparticles; Flash nanocomplexation; Insulin; Oral delivery; Scalable fabrication