Preparation and characterization of SDF-1α-chitosan-dextran sulfate nanoparticles

J Vis Exp. 2015 Jan 22:(95):52323. doi: 10.3791/52323.

Abstract

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under appropriate conditions. The glycan nanoparticles are useful carriers for protein factors, which facilitate the in vivo delivery of the proteins and sustain their retention in the targeted tissue. The glycan polyelectrolyte complexes are also ideal for protein delivery, as the incorporation is carried out in aqueous solution, which reduces the likelihood of inactivation of the proteins. Proteins with a heparin-binding site adhere to dextran sulfate readily, and are, in turn, stabilized by the binding. These particles are also less inflammatory and toxic when delivered in vivo. In the protocol described below, SDF-1α (Stromal cell-derived factor-1α), a stem cell homing factor, is first mixed and incubated with dextran sulfate. Chitosan is added to the mixture to form polyelectrolyte complexes, followed by zinc sulfate to stabilize the complexes with zinc bridges. The resultant SDF-1α-DS-CS particles are measured for size (diameter) and surface charge (zeta potential). The amount of the incorporated SDF-1α is determined, followed by measurements of its in vitro release rate and its chemotactic activity in a particle-bound form.

Publication types

  • Research Support, N.I.H., Extramural
  • Video-Audio Media

MeSH terms

  • Chemokine CXCL12 / chemistry*
  • Chemotaxis / drug effects
  • Chitosan / chemistry*
  • Dextran Sulfate / chemistry*
  • Heparin / chemistry
  • Humans
  • Jurkat Cells
  • Nanoparticles / chemistry*
  • Particle Size

Substances

  • Chemokine CXCL12
  • chitosan sulfate
  • Heparin
  • Chitosan
  • Dextran Sulfate