Influence of charge on FITC-BSA-loaded chondroitin sulfate-chitosan nanoparticles upon cell uptake in human Caco-2 cell monolayers

Abstract

Background and methods:Chondroitin sulfate-chitosan (ChS-CS) nanoparticles and positively and negatively charged fluorescein isothiocyanate-conjugated bovine serum albumin (FITC-BSA)-loaded ChS-CS nanoparticles were prepared and characterized. The properties of ChS-CS nanoparticles, including cellular uptake, cytotoxicity, and transepithelial transport, as well as findings on field emission-scanning electron microscopy, transmission electron microscopy, and confocal laser scanning microscopy were evaluated in human epithelial colorectal adenocarcinoma (Caco-2) fibroblasts. ChS-CS nanoparticles with a mean particle size of 250 nm and zeta potentials ranging from −30 to +18 mV were prepared using an ionic gelation method. Results: Standard cell viability assays demonstrated that cells incubated with ChS-CS and FITC-BSA-loaded ChS-CS nanoparticles remained more than 95% viable at particle concentrations up to 0.1 mg/mL. Endocytosis of nanoparticles was confirmed by confocal laser scanning microscopy and measured by flow cytometry. Ex vivo transepithelial transport studies using Caco-2 cells indicated that the nanoparticles were effectively transported into Caco-2 cells via endocytosis. The uptake of positively charged FITC-BSA-loaded ChS-CS nanoparticles across the epithelial membrane was more efficient than that of the negatively charged nanoparticles. Conclusion:The ChS-CS nanoparticles fabricated in this study were effectively endocytosed by Caco-2 fibroblasts without significant cytotoxicity at high nanoparticle concentrations. ChS-CS nanoparticles represent a potential novel delivery system for the transport of hydrophilic macromolecules.

Chitosan-coated mesoporous MIL-100(Fe) nanoparticles as improved bio-compatible oral nanocarriers

“…5 ). This is in agreement with previous described results using CS-coated NPs of different nature 19 41 , in which the bioadhesive capacity of CS 34 is associated with the increase of NPs cell penetration through an improvement of NP-cell contact 41 42 . To further assess whether these NPs were internalized into the cells, confocal microscopy images were collected at different levels of the Z-axis ( Figures S10–S12 ), detecting the iron self-reflection fluorescence coming from the nanoMOFs at different depths.…”

“…Finally, the quantification of the linker of the NPs in the basolateral compartment was analyzed by HPLC (see methods), whereas the NP internalization into the cell monolayers after the incubation times were examined by confocal fluorescence microscopy, described in the section below. Each experiment was performed in duplicate 42 49 50 .…”

Chitosan Based Self-Assembled Nanoparticles in Drug Delivery

“…Its biological properties have stimulated the preparation and evaluation of CS-CHOS nanoparticles for drug/gene delivery [117,118] and delivery of platelet lysates [119]. CS-CHOS nanoparticles have been suggested as a novel delivery system for the transport of hydrophilic macromolecules [120].…”

Novel Anthra[1,2-c][1,2,5]Thiadiazole-6,11-Diones as Promising Anticancer Lead Compounds: Biological Evaluation, Characterization & Molecular Targets Determination

“…Cell monolayers with TEER values less than 300 Ω.cm 2 at the end of the maintenance period were not suitable for the experiment and discarded [ 43 ]. The integrity of the cell monolayers were further confirmed by testing the phenol red permeability through the cells [ 55 ]. The transport experiments were started by replacing the incubation culture media with the transport buffers with or without the tested compounds, the transport was tested in both directions (apical to basolateral and basolateral to apical).…”