title: Depolymerized chitosans functionalized with bPEI as carriers of nucleic acids and tuftsin-tethered conjugate for macrophage targeting.
abstract: Development of efficient and safe nucleic acid carriers (vectors) is one of the essential requirements for the success of gene therapy. Here, we have evaluated the gene transfer capability of chitosan-PEI (CP) conjugates prepared by conjugating low molecular weight branched polyethylenimine (LMWP) with depolymerized chitosans (7 and 10 kDa) via their terminal aldehyde/keto groups. The CP conjugates interacted efficiently with nucleic acids and also showed higher cellular uptake. These conjugates on complexation with DNA yielded nanoparticles in the size range of 100-130 nm (in case of C7P) and 115-160 nm (in case of C10P), which exhibited significantly higher transfection efficiency (~2-42 folds) in vitro compared to chitosans (high and low mol. wt.) and the commercially available transfection reagents retaining cell viability almost comparable to the native chitosan. Of the two CP conjugates, chitosan 7 kDa-LMWP (C7P) displayed higher gene transfer ability in the presence and absence of serum. Luciferase reporter gene analysis in male Balb/c mice receiving intravenous administration of C7P3/DNA polyplex showed the maximum expression in their spleen. Further, tuftsin, a known macrophage targeting molecule, was tethered to C7P3 and the resulting complex, i.e., C7P3-T/DNA, exhibited significantly higher gene expression in cultured mouse peritoneal macrophages as compared to unmodified C7P3/DNA complex without any cytotoxicity demonstrating the suitability of the conjugate for targeted applications. Conclusively, the study demonstrates the potential of the projected conjugates for gene delivery for wider biomedical applications.
authors.affiliation: CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Delhi 110007, India.
authors.forename: Sushil K
authors.initials: SK
authors.lastname: Tripathi
authors.forename: Ritu
authors.initials: R
authors.lastname: Goyal
authors.forename: Mahendra P
authors.initials: MP
authors.lastname: Kashyap
authors.forename: Aditya B
authors.initials: AB
authors.lastname: Pant
authors.forename: Wahajul
authors.initials: W
authors.lastname: Haq
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kumar
authors.forename: Kailash C
authors.initials: KC
authors.lastname: Gupta
mesh_terms: D000818:Animals; D002460:Cell Line; D048271:Chitosan / Q000737:chemistry*; D005434:Flow Cytometry; D006801:Humans; D008264:Macrophages / Q000187:drug effects*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D009696:Nucleic Acids / Q000008:administration & dosage / Q000494:pharmacology*; D010316:Particle Size; D011094:Polyethyleneimine; D011108:Polymers / Q000737:chemistry*; D034741:RNA, Small Interfering; D014405:Tuftsin / Q000737:chemistry*
chemical_list: D009696:Nucleic Acids; D011108:Polymers; D034741:RNA, Small Interfering; D011094:Polyethyleneimine; D048271:Chitosan; D014405:Tuftsin