Glutaraldehyde crosslinked and alkaline denaturation induced self association of haemoglobin to design nanocarriers for In vitro release of insulin in simulated gastrointestinal fluids (SGFs)

Abstract

Adopting a novel synthetic route the haemoglobin nanoparticles were prepared by its simultaneous microemulsion mediated crosslinking with glutaraldehyde and alkaline induced denaturation. The as prepared nanoparticles were characterized by techniques like Fourier transform infrared spectroscopy (FTIR), Transmission electron microscopy (TEM), Scanning electron microcopy (SEM), Zeta potential and Dynamic Light Scattering measurements. The particle size analysis revealed that size of the nanoparticles lays in the range 60–150 nm with surface charge of −27.1 mV. The TEM images suggested for aggregated structures which confirm denaturation of haemoglobin macromolecules under alkaline conditions. The nanoparticles were assessed for water intake capacity and the effect of various factors like chemical composition of nanoparticles, pH and temperature of the swelling bath, and simulated biological fluids on water sorption capacity was investigated. The insulin was loaded on to the prepared nanoparticles and investigated for swelling controlled release of insulin in simulated gastrointestinal fluid (SGIFs). The influence of chemical composition of nanoparticles, pH and temperature of the release media, and simulated physiological fluids was studied on the released amount of insulin and an optimized formulation was achieved. The prepared haemoglobin nanoparticles were also investigated for their in-vitro cytotoxicity.