title: Mannosylated chitosan nanoparticles for delivery of antisense oligonucleotides for macrophage targeting.
abstract: The therapeutic potential of antisense oligonucleotides (ASODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study focuses on designing mannosylated low molecular weight (LMW) chitosan nanoconstructs for safe ODNs delivery by macrophage targeting. Mannose groups were coupled with LMW chitosan and characterized spectroscopically. Mannosylated chitosan ODN nanoparticles (MCHODN NPs) were formulated by self-assembled method using various N/P ratio (moles of amine groups of MCH to phosphate moieties of ODNs) and characterized for gel retardation assay, physicochemical characteristics, cytotoxicity and transfection efficiency, and antisense assay. Complete complexation of MCH/ODN was achieved at charge ratio of 1:1 and above. On increasing the N/P ratio of MCH/ODN, particle size of the NPs decreased whereas zeta potential (ZV) increased. MCHODN NPs displayed much higher transfection efficiency into Raw 264.7 cells (bears mannose receptors) than Hela cells and no significant toxicity was observed at all MCH concentrations. Antisense assay revealed that reduction in lipopolysaccharide (LPS) induced serum TNF-α is due to antisense activity of TJU-2755 ODN (sequence complementary to 3'-UTR of TNF-α). These results suggest that MCHODN NPs are acceptable choice to improve transfection efficiency in vitro and in vivo.
authors: Maharishi Markandeshwar College of Pharmacy, Maharishi Markandeshwar University, Mullana Ambala, Haryana 133207, India ; Department of Pharmaceutical Sciences, Dr. Hari Singh Gour Vishwa Vidyala, Sagar, Madhya Pradesh 470003, India.
authors: Gyati Shilakari
authors: 0000-0002-2895-2332
authors: GS
authors: Asthana
authors: Maharishi Markandeshwar College of Pharmacy, Maharishi Markandeshwar University, Mullana Ambala, Haryana 133207, India.
authors: Abhay
authors: A
authors: Asthana
authors: Department of Pharmaceutical Sciences, Dr. Hari Singh Gour Vishwa Vidyala, Sagar, Madhya Pradesh 470003, India.
authors: Dharm Veer
authors: DV
authors: Kohli
authors: Department of Pharmaceutical Sciences, Dr. Hari Singh Gour Vishwa Vidyala, Sagar, Madhya Pradesh 470003, India.
authors: Suresh Prasad
authors: SP
authors: Vyas
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D048271:Chitosan / Q000737:chemistry*; D003850:Deoxyribonuclease I / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry*; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D008358:Mannose / Q000737:chemistry*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008970:Molecular Weight; D053758:Nanoparticles / Q000737:chemistry*; D016376:Oligonucleotides, Antisense / Q000737:chemistry*; D010316:Particle Size; D012995:Solubility; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014867:Water / Q000737:chemistry
chemical_list: D004337:Drug Carriers; D016376:Oligonucleotides, Antisense; D014409:Tumor Necrosis Factor-alpha; D014867:Water; D048271:Chitosan; D003850:Deoxyribonuclease I; D008358:Mannose