A Full Set of In Vitro Assays in Chitosan/Tween 80 Microspheres Loaded with Magnetite Nanoparticles

Abstract

Microspheres have been proposed for different medical applications, such as the delivery of therapeutic proteins. The first step, before evaluating the functionality of a protein delivery system, is to evaluate their biological safety. In this work, we developed chitosan/Tween 80 microspheres loaded with magnetite nanoparticles and evaluated cell damage. The formation and physical–chemical properties of the microspheres were determined by FT-IR, Raman, thermogravimetric analysis (TGA), energy-dispersive X-ray spectroscopy (EDS), dynamic light scattering (DLS), and SEM. Cell damage was evaluated by a full set of in vitro assays using a non-cancerous cell line, human erythrocytes, and human lymphocytes. At the same time, to know if these microspheres can load proteins over their surface, bovine serum albumin (BSA) immobilization was measured. Results showed 7 nm magnetite nanoparticles loaded into chitosan/Tween 80 microspheres with average sizes of 1.431 µm. At concentrations from 1 to 100 µg/mL, there was no evidence of changes in mitochondrial metabolism, cell morphology, membrane rupture, cell cycle, nor sister chromatid exchange formation. For each microgram of microspheres 1.8 µg of BSA was immobilized. The result provides the fundamental understanding of the in vitro biological behavior, and safety, of developed microspheres. Additionally, this set of assays can be helpful for researchers to evaluate different nano and microparticles.

Modification of Branched Polyethyleneimine Using Mesquite Gum for Its Improved Hemocompatibility

“…This lysis produces the release of the intracellular content of the erythrocyte due to the rupture of its membrane. The released molecule measured was hemoglobin, which is a predominant protein in erythrocytes [ 47 ]. To the best of our knowledge, there are few works that give a report on the hemocompatibility of natural gums, in the case of the work by Goyal et al [ 23 ], they did not carry out hemolysis testing.…”

“…A hemolysis test was developed to determine the effect of polymers on erythrocyte lysis. The protocol followed in the present work was based on the investigation reported by Roacho et al with some modifications [ 47 ].…”

Carboxymethylated Gums and Derivatization: Strategies and Significance in Drug Delivery and Tissue Engineering

“…This polymer has high charge density, the potential to destabilize membranes, and the capacity to shield endocytosed plasmid DNA from enzymatic deterioration, rendering it ideal for encasing plasmid DNA in nanoparticles for gene therapy. PEI’s toxicity increases with molecule weight and thus is considered a risk for gene delivery [ 114 , 115 ]. These polymers do not break down in the body and pose a long-term threat [ 116 ].…”

Green Synthesis of Mesquite-Gum-Stabilized Gold Nanoparticles for Biomedical Applications: Physicochemical Properties and Biocompatibility Assessment

“…Hematoxylin and eosin (H&E) stain was used to evaluate the morphology of the fibroblast 3T3L1 cells, which had been exposed to different concentrations of AuNPs@MG (4.5, 1.11, and 0.28 µg/mL); for comparison, cells were also exposed to the same concentrations of MG. A previously reported protocol was followed [31]. First, the culture medium was carefully extracted, and cells were PBS-buffer-washed (Thermo Fisher Scientific, Waltham, MA, USA) three times.…”