title: Incorporation of heparin-binding proteins into preformed dextran sulfate-chitosan nanoparticles.
abstract: Incorporation of proteins into dextran sulfate (DS)-chitosan (CS) nanoparticles (DSCS NPs) is commonly performed using entrapment procedures, in which protein molecules are mixed with DS and CS until particle formation occurs. As DS is an analog of heparin, the authors examined whether proteins could be directly incorporated into preformed DSCS NPs through a heparin binding domain-mediated interaction. The authors formulated negatively-charged DSCS NPs, and quantified the amount of charged DS in the outer shell of the particles. The authors then mixed the DSCS NPs with heparin-binding proteins (SDF-1α, VEGF, FGF-2, BMP-2, or lysozyme) to achieve incorporation. Data show that for DSCS NPs containing 100 nmol charged glucose sulfate units in DS, up to ~1.5 nmol of monomeric or ~0.75 nmol of dimeric heparin-binding proteins were incorporated without significantly altering the size or zeta potential of the particles. Incorporation efficiencies of these proteins were 95%-100%. In contrast, serum albumin or serum globulin showed minimal incorporation (8% and 4%, respectively) in 50% physiological saline, despite their large adsorption in water (80% and 92%, respectively). The NP-incorporated SDF-1α and VEGF exhibited full activity and sustained thermal stability. An in vivo aerosolization study showed that NP-incorporated SDF-1α persisted in rat lungs for 72 h (~34% remaining), while free SDF-1α was no longer detectable after 16 h. As many growth factors and cytokines contain heparin-binding sites/domains, incorporation into preformed DSCS NPs could facilitate in vivo applications of these proteins.
authors.affiliation: Department of Medicine, Brigham and Women's Hospital.
authors.forename: Paula
authors.initials: P
authors.lastname: Zaman
authors.affiliation: Department of Medicine, Brigham and Women's Hospital.
authors.forename: Julia
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Medicine, Brigham and Women's Hospital.
authors.forename: Adam
authors.initials: A
authors.lastname: Blau
authors.affiliation: Department of Anesthesiology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.
authors.forename: Weiping
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Medicine, Brigham and Women's Hospital.
authors.forename: Tina
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Anesthesiology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Kohane
authors.affiliation: Department of Medicine, Brigham and Women's Hospital.
authors.forename: Joseph
authors.initials: J
authors.lastname: Loscalzo
authors.affiliation: Department of Medicine, Brigham and Women's Hospital.
authors.forename: Ying-Yi
authors.initials: YY
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D055396:Bone Morphogenetic Protein 2 / Q000737:chemistry / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D002626:Chemistry, Pharmaceutical; D054377:Chemokine CXCL12 / Q000737:chemistry / Q000378:metabolism; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism*; D016264:Dextran Sulfate / Q000737:chemistry / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry / Q000378:metabolism; D006493:Heparin / Q000737:chemistry / Q000378:metabolism*; D008168:Lung / Q000737:chemistry / Q000378:metabolism; D008297:Male; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry / Q000378:metabolism
chemical_list: D001798:Blood Proteins; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D054377:Chemokine CXCL12; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D016222:Fibroblast Growth Factor 2; D006493:Heparin; D048271:Chitosan; D016264:Dextran Sulfate; D009113:Muramidase
keywords: glycan; glycosaminoglycan; polyelectrolyte complex; polysaccharide