title: Investigation of the cytokine response to NF-κB decoy oligonucleotide coated polysaccharide based nanoparticles in rheumatoid arthritis in vitro models.
abstract: BACKGROUND
  The transcription factor nuclear factor-kappa B (NF-κB) is highly involved in regulation of a number of cellular processes, including production of inflammatory mediators. Thus, this transcription factor plays a role in pathology of many diseases, including rheumatoid arthritis, an autoimmune disease hallmarked by an imbalance of pro and anti-inflammatory cytokines. Small nucleic acids with sequences that mimic the native binding site of NF-κB have been proposed as treatment options for RA; however due to low cellular penetration and a high degree of instability, clinical applications of these therapeutics have been limited.
  
  
  METHODS
  Here, we describe the use of N-trimethyl chitosan-polysialic acid (PSA-TMC) nanoparticles coated with decoy oligodeoxynucleotides (ODNs) specific to transcription factor NF-κB (PSA-TMC-ODN) as a method to enhance the stability of the nucleic acids and facilitate increased cellular penetration. In addition to decoy ODN, PSA-TMC nanoparticles were loaded with RA therapeutic methotrexate (MTX), to assess the anti-inflammatory efficacy of a combination therapy approach. Two different in vitro models, a cell line based model as well as a primary RA cell model were used to investigate anti-inflammatory activity. One way ANOVA followed by Holm-Sidak stepdown comparisons was used to determine statistical significance.
  
  
  RESULTS
  In general, free ODN did not significantly affect secretion of pro-inflammatory cytokines interleukin-6 (IL-6) and interleukin-8, (IL-8) while free MTX had variable efficacy. However, PSA-TMC-ODN and PSA-TMC-ODN-MTX resulted in significant decreases in the inflammatory mediators IL-6 and IL-8 in both cell models. In addition, PSA-TMC exhibited sufficient cellular uptake, as observed through fluorescence microscopy.
  
  
  CONCLUSIONS
  These results support our previous findings that PSA-TMC nanoparticles are an effective delivery vehicle for small nucleic acids, and effectively alter the pro-inflammatory state characteristic of RA.
authors.affiliation: Syracuse Biomaterials Institute, Syracuse University, 318 Bowne Hall, Syracuse, NY, 13244, USA. prwardwe@syr.edu.|Department of Biomedical and Chemical Engineering, Syracuse University, 121 Link Hall, Syracuse, NY, 13244, USA. prwardwe@syr.edu.
authors.forename: Patricia R
authors.initials: PR
authors.lastname: Wardwell
authors.affiliation: Syracuse Biomaterials Institute, Syracuse University, 318 Bowne Hall, Syracuse, NY, 13244, USA.|Department of Physics, Syracuse University, Syracuse, NY, 13244, USA.
authors.forename: Martin B
authors.initials: MB
authors.lastname: Forstner
authors.affiliation: Syracuse Biomaterials Institute, Syracuse University, 318 Bowne Hall, Syracuse, NY, 13244, USA. babader@syr.edu.|Department of Biomedical and Chemical Engineering, Syracuse University, 121 Link Hall, Syracuse, NY, 13244, USA. babader@syr.edu.
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Bader
mesh_terms: D018501:Antirheumatic Agents / Q000008:administration & dosage*; D001172:Arthritis, Rheumatoid / Q000276:immunology*; D002460:Cell Line; D048271:Chitosan; D002851:Chromatography, High Pressure Liquid; D016207:Cytokines / Q000096:biosynthesis*; D004337:Drug Carriers; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D066298:In Vitro Techniques; D008727:Methotrexate / Q000008:administration & dosage*; D053758:Nanoparticles / Q000008:administration & dosage*; D009838:Oligodeoxyribonucleotides / Q000276:immunology / Q000494:pharmacology*; D009841:Oligonucleotides; D011134:Polysaccharides; D012794:Sialic Acids
chemical_list: D018501:Antirheumatic Agents; D016207:Cytokines; D004337:Drug Carriers; C118071:N-trimethyl chitosan chloride; C468119:NF-kappaB decoy; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D011134:Polysaccharides; D012794:Sialic Acids; C021319:polysialic acid; D048271:Chitosan; D008727:Methotrexate