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Enhancement of antimicrobial activity of chlorhexidine by liposomes with chitosan

Chronic wounds affect 1-2% of the population. In the presented study, chitosan-coated and chitosan-containing liposomes were prepared to enhance the antibacterial effect of the drug chlorhexixdine. A chitosan-based delivery system enhances the antimicrobial activity of chlorhexidine.


Hemmingsen LM, Panchai P, Julin K, Basnet P, Nystad M, Johannessen M, Škalko-Basnet N. Chitosan-based delivery system enhances antimicrobial activity of chlorhexidine. Front Microbiol. 2022 Sep 29;13:1023083. doi: 10.3389/fmicb.2022.1023083. PMID: 36246245; PMCID: PMC9557914.

Chronic wounds are a burden on the healthcare system. In 2018, it is roughly estimated that 1-2% of the population is affected. Microbial infections and inflammations are the main factors that prevent wounds from healing. In addition, the number of microbial resistances is growing, where biofilm formation in particular poses a major threat. Therefore, innovations are urgently needed to improve the situation.

Chitosan and liposomes are considered to be particularly promising. Chitosan has antibacterial and anti-inflammatory properties and is anti-oxidative and hemostatic. Lipid-based active ingredient transporters such as liposomes are able to interact with the skin and delay the release of active ingredients. This allows for higher drug concentrations at the wound site. In this study, the potential of chitosan-coated and chitosan-containing liposomes was investigated. For this purpose, chitosan was coupled with chlorhexidine, an antibacterial agent, to achieve a synergistic effect. A chitosan with a low molecular weight (50-1000 kDa, DDA ˃ 70%) was selected for this purpose. The prepared liposomes with chitosan were characterized and evaluated for in vitro chlorhexidine release, cytotoxicity, and antimicrobial properties. In addition to adding chlorhexidine to chitosan-coated and chitosan-containing vesicles, chlorhexidine was also added to chitosan-free vesicles.


  • Successful synthesis of vesicles with a size of 250 and 350 nm and a PI smaller than 0.4
  • Long-term stability of the vesicles of up to 4 weeks for PI, size, zeta potential and pH
  • Prolongation of the in vitro release of chlorhexidine in the chitosan-containing vesicles
  • No observation of cytotoxicity toward HaCaT and NHDF-neo cells regardless of vesicle type
  • Inflammatory responses in murine macrophages treated with vesicles were reduced by approximately 60% compared to untreated cells
  • Dose-dependent, inflammation-inhibiting effect with chitosan and chitosan-chlorhexidine liposomes
  • with chitosan in the vesicles, bacterial survival was drastically reduced to 5.3 and 4.7% for the chitosan-containing and chitosan-coated liposomes, respectively
  • Vesicles containing chitosan and chlorhexidine in combination showed the strongest effects against S. aureus

Summary: In the presented study, it was shown that lipid-based vesicles, such as liposomes, with chitosan can be promising transporters for antibacterial agents such as chlorhexidine. Due to chitosan, these vesicles showed enhanced antibacterial activity against S. aureus.

Link to article: Frontiers | Chitosan-based delivery system enhances antimicrobial activity of chlorhexidine (

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