Effect of Lactoferrin Loaded on Chitosan Against Staphylococcus Aureus and Pseudomonas Aeruginosa

Abstract

The increasing resistance of bacterial strains to antimicrobial compounds presents a significant global health challenge. Staphylococcus aureus and Pseudomonas aeruginosa are two such bacteria that pose substantial risks, particularly in causing infections of the respiratory tract and bacteremia. Chitosan, a non-toxic and biocompatible polymer, has demonstrated inhibitory effects on bacterial growth. Lactoferrin, an iron-binding protein found in milk, exhibits antimicrobial properties. In this study, lactoferrin was purified from camel milk using CM Sephadex C-50 chromatography after removing casein through ion exchange chromatography. The purification process was confirmed using SDS-PAGE and the absence of color with tetramethylbenzidine. Subsequently, purified lactoferrin was loaded onto 1% chitosan using thiamine pyrophosphate (TPP). The successful loading of lactoferrin onto chitosan was verified using Scanning Electron Microscopy (SEM), Zeta Potential, Particle Size Determination, and Fourier Transform Infrared Spectroscopy (FTIR). After evaluating the cytotoxicity of various lactoferrin concentrations loaded on chitosan, the antimicrobial effects of lactoferrin (at concentrations of 300 and 350 μg/mL) loaded on chitosan were assessed using a microassay method. The results demonstrated a significant decrease (P-value < 0.01) in the growth of Staphylococcus aureus and Pseudomonas aeruginosa bacteria compared to the control group when treated with 300 μg/mL of lactoferrin-loaded chitosan. Furthermore, a substantial decrease (P-value < 0.001) was observed with a concentration of 350 μg/mL.