Development and characterization of glutathione-conjugated albumin nanoparticles for improved brain delivery of hydrophilic fluorescent marker

Abstract

The glutathione-conjugated bovine serum albumin (BSA) nanoparticles were constructed in the present exploration as a novel biodegradable carrier for brain-specific drug delivery with evaluation of its in vitro and in vivo delivery properties. BSA nanocarriers were activated and conjugated to the distal amine functions of the glutathione via carbodiimide chemistry using EDAC as a mediator. These nanoparticles were characterized for particle shape, average size, SPAN value, drug entrapment and in vitro drug release. Further, presence of glutathione on the surface of BSA nanoparticles was confirmed by Ellman's assay, which has suggested that approximately 750 units of glutathione were conjugated per BSA nanoparticle. To evaluate the brain delivery properties of the glutathione-conjugated BSA nanoparticles fluorescein sodium was used as a model hydrophilic compound. Permeability and neuronal uptake properties of developed formulations were evaluated against the MDCK-MDR1 endothelial and neuro-glial cells, respectively. The permeability of glutathione-conjugated BSA nanoparticles across the monolayer of MDCK-MDR1 endothelial tight junction was shown significantly higher than that of unconjugated nanoparticles and fluorescein sodium solution. Similarly, glutathioneconjugated nanoparticles exhibited considerably higher uptake by neuro-glial cells which was inferred by high fluorescence intensity under microscope in comparison to unconjugated nanoparticles and fluorescein sodium solution. Following an intravenous administration, nearly three folds higher fluorescein sodium was carried to the rat brain by glutathione-conjugated nanoparticles as compared to unconjugated nanoparticles. The significant in vitro and in vivo results suggest that glutathione-conjugated BSA nanoparticles is a promising brain drug delivery system with low toxicity.

Combination of Alanine and Glutathione as Targeting Ligands of Nanoparticles Enhances Cargo Delivery into the Cells of the Neurovascular Unit

“…This 2.2-fold increase in cargo delivery is in agreement with a recent study in which GSH-functionalized polymeric NPs were able to deliver curcumin with comparable efficacy into SK-N-SH neuronal cells, a maternal cell line of SH-SY5Y [ 68 ]. Moreover, an increase in the uptake of fluorescein-loaded albumin NPs in a mixed neuron-glia co-culture were described when the particles were labeled with glutathione [ 69 ]. We also observed an increase in cargo delivery by our dual-labeled NPs, while cargo fluorescence was barely detectable in SH-SY5Y cells treated with either free cargo or non-targeted niosomes ( Figure 7 c).…”

Glutathione: Antioxidant Properties Dedicated to Nanotechnologies

“…In animals, bovine serum albumin (BSA) NPs with a GSH ligand led to 10-fold higher brain concentrations of a neuroprotective agent than the drug solution, 5 h after intravenous injection [ 153 ]. In another study, a hydrophilic model drug was 3 times more concentrated in the brain after intravenous injection of GSH-coated BSA NPs, compared to the same uncoated NPs [ 154 ]. Finally, in a middle cerebral artery occlusion model of stroke, GSH-coated PLGA–PEG NPs containing the thyroid T3 hormone showed better therapeutic efficacy than T3 solution or uncoated NPs, on both tissue infarction (reduction of 58%, 34%, and 51%, respectively) and brain edema (reduction of 75%, 59% and 68%, respectively) [ 155 ].…”

Fabrication of peptide-linked albumin nanoconstructs for receptor-mediated delivery of asiatic acid to the brain as a preventive measure in cognitive impairment: optimization, in-vitro and in-vivo evaluation

“…BAG nps were prepared by carbodiimide reaction using EDAC as linker ( Figure 1) (refer Supplementary data for detailed procedure) [10,13].…”

“…Preliminary research findings by Patel et al reveal that GU-conjugated fluorescein sodium labelled bovine serum albumin (BSA) nps showed 9-fold improvement in C max of fluorescein sodium in brain was achieved compared to fluorescein sodium solution. Subsequently, GU-conjugated paclitaxel loaded PLGA nps depicted 6.1-fold improved as compared to paclitaxel suspension [10][11][12]. Based on this data, preliminary studies were carried out to formulate GU-conjugated albumin nps of AA wherein 3.2-fold improvement in C max of AA in the brain was achieved as compared to AA dispersion [13].…”