Characterization of Sulfated HA Nanoparticles

Hydrodynamic diameter (HD) and zeta potential measurements of all sulfated derivatives of HA NPs were measured by dynamic light scattering (DLS) using ZetaSizer NanoZS90 (Malvern Instruments; Malvern UK). NPs samples were prepared by dissolving samples in ultrapure water (lmg/mL) and filtered through 0.45 pm syringe filter. Absorbance spectra of the NPs were recorded with UV-2600 spectrometer (ThermoFisher Evolution 220) in water and 1:1 (v/v) mixture of DMS0:H₂0. Fluorescence reads were obtained on a FluoroMax-4 fluorescence spectrometer equipped with a NIR extended range PMT (Horiba Jobin Yvon; Edison, NJ, USA). Conjugation ratio of Cy7.5 dye to HA was determined using standard curve developed for Cy7.5 dye in 1:1 (v/v) DMS0:H₂0. Conjugation of the hydrophobic ligand was confirmed by ¹H-NMR for all the derivatives of sulfated HA-PBA. Transmission electron microscope (TEM) images were obtained for NPs (1 mg/mL concentration), placed on formavar/silicone monoxide-coated 200 mesh copper grids using NanoVan negative stain for ^{33 (link)–34 (link)} s and imaged using the FEI Tecnai G2 Spirit TEM (FEI; Hillsboro, Oregon) at UNMC's electron microscopy core facility.

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Bhattacharya D.S., Svechkarev D., Bapat A., Patil P., Hollingsworth M.A, & Mohs A.M. (2020). Sulfation modulates the targeting properties of hyaluronic acid to P-selectin and CD44. ACS biomaterials science & engineering, 6(6), 3585-3598.

Publication 2020

ZetaSizer NanoZS90 Evolution 220 NIR extended range PMT Tecnai G2 Spirit TEM

1h nmr Copper Cy7 dye Derivatives Electron microscopy Evolution Fluorescence Hydrodynamic Ligand Silicone Stain Syringe Transmission electron microscope

Corresponding Organization : University of Nebraska Medical Center

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Variable analysis

independent variables

Hydrodynamic diameter (HD) and zeta potential of sulfated derivatives of HA NPs

dependent variables

Absorbance spectra of the NPs in water and 1:1 (v/v) DMSO:H2O
Fluorescence reads of the NPs
Conjugation ratio of Cy7.5 dye to HA
Conjugation of the hydrophobic ligand confirmed by 1H-NMR
Transmission electron microscope (TEM) images of the NPs

control variables

Concentration of NP samples (1 mg/mL)
Solvent used for NP preparation (ultrapure water)
Filtration of NP samples through 0.45 μm syringe filter

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