Incorporation of heparin-binding proteins into preformed dextran sulfate-chitosan nanoparticles

• Published in: International journal of nanomedicine Vol. 11; pp. 6149 - 6159
• Main Authors: Zaman, Paula, Wang, Julia, Blau, Adam, Wang, Weiping, Li, Tina, Kohane, Daniel S, Loscalzo, Joseph, Zhang, Ying-Yi
• Format: Journal Article
• Language: English
• Published: New Zealand Dove Medical Press Limited 01.01.2016; Taylor & Francis Ltd; Dove Medical Press
• Subjects: Animals; Anticoagulants; Binding proteins; Binding sites; Blood Proteins - chemistry; Blood Proteins - metabolism; Bone Morphogenetic Protein 2 - chemistry; Bone Morphogenetic Protein 2 - metabolism; Cell Movement; Cell Proliferation; Chemistry, Pharmaceutical; Chemokine CXCL12 - chemistry; Chemokine CXCL12 - metabolism; Chitosan - chemistry; Chitosan - metabolism; Dextran; Dextran Sulfate - chemistry; Dextran Sulfate - metabolism; Fibroblast Growth Factor 2 - chemistry; Fibroblast Growth Factor 2 - metabolism; Fibroblasts; glycan; glycosaminoglycan; Glycosaminoglycans; Heparan sulfate; Heparin - chemistry; Heparin - metabolism; Laboratories; Lung - chemistry; Lung - metabolism; Lysozyme; Male; Muramidase - chemistry; Muramidase - metabolism; Nanoparticles; Nanoparticles - chemistry; Nanoparticles - metabolism; Original Research; polyelectrolyte complex; polysaccharide; Protein binding; Proteins; Rats; Rats, Sprague-Dawley; Sulfates; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - chemistry; Vascular Endothelial Growth Factor A - metabolism; glycan; polyelectrolyte complex; polysaccharide; glycosaminoglycan
• ISSN: 1178-2013; 1176-9114; 1178-2013
• DOI: 10.2147/IJN.S119174

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.