title: Effects of tripolyphosphate on cellular uptake and RNA interference efficiency of chitosan-based nanoparticles in Raw 264.7 macrophages.
abstract: Tumor necrosis factor-α (TNF-α) is a major pro-inflammatory cytokine that is mainly secreted by macrophages during inflammation. Here, we synthesized a series of N-(2-hydroxy)propyl-3-trimethyl ammonium chitosan chlorides (HTCCs), and then used a complex coacervation technique or tripolyphosphate (TPP)-assisted ionotropic gelation strategy to complex the HTCCs with TNF-α siRNA (siTNF) to form nanoparticles (NPs). The resultant NPs had a desirable particle size (210-279nm), a slightly positive zeta potential (14-22mV), and negligible cytotoxicity against Raw 264.7 macrophages and colon-26 cells. Subsequent cellular uptake tests demonstrated that the introduction of TPP to the NPs markedly increased their cellular uptake efficiency (to nearly 100%) compared with TPP-free NPs, and yielded a correspondingly higher intracellular concentration of siRNA. Critically, in vitro gene silencing experiments revealed that all of the TPP-containing NPs showed excellent efficiency in inhibiting the mRNA expression level of TNF-α (by approximately 85-92%, which was much higher than that obtained using Oligofectamine/siTNF complexes). Collectively, these results obviously suggest that our non-toxic TPP-containing chitosan-based NPs can be exploited as efficient siTNF carriers for the treatment of inflammatory diseases.
authors.affiliation: Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, PR China; Institute for Biomedical Science, Center for Diagnostics and Therapeutics, Georgia State University, Atlanta 30302, USA. Electronic address: bxiao@gsu.edu.
authors.forename: Bo
authors.initials: B
authors.lastname: Xiao
authors.affiliation: Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, PR China.
authors.forename: Panpan
authors.initials: P
authors.lastname: Ma
authors.affiliation: Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, PR China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Ma
authors.affiliation: Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, PR China.
authors.forename: Qiubing
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, PR China.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Si
authors.affiliation: Institute for Biomedical Science, Center for Diagnostics and Therapeutics, Georgia State University, Atlanta 30302, USA.
authors.forename: Lewins
authors.initials: L
authors.lastname: Walter
authors.affiliation: Institute for Biomedical Science, Center for Diagnostics and Therapeutics, Georgia State University, Atlanta 30302, USA; Atlanta Veterans Affairs Medical Center, Decatur 30033, USA.
authors.forename: Didier
authors.initials: D
authors.lastname: Merlin
mesh_terms: D000818:Animals; D048271:Chitosan / Q000031:analogs & derivatives* / Q000737:chemistry; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry*; D011122:Polyphosphates / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D000067996:RAW 264.7 Cells; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000493:pharmacokinetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*
chemical_list: C550160:N-(2-hydroxypropyl)-3-trimethylammonium chitosan; D011122:Polyphosphates; D000644:Quaternary Ammonium Compounds; D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha; D048271:Chitosan; C005692:triphosphoric acid
keywords: Chitosan; Macrophage; N-(2-hydroxy)propyl-3-trimethyl ammonium chitosan chloride; Nanoparticle; RNA interference; Tripolyphosphate