A New Amphotericin B-loaded Trimethyl Chitosan Nanoparticles as a Drug Delivery System and Antifungal Activity on Candida albicans Biofilm

Document Type : Original Articles


1 Department of Microbiology, Medical and Veterinary Mycology, Faculty of Veterinary Specialized Science, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Human Vaccine and Serum, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

3 Department of Tuberculosis, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran


Amphotericin B (AmB) is an effective antifungal agent; however, the application of AmB is associated with a number of drawbacks. Application of nanoparticles (NPs) is known to improve the efficiency of drug delivery to the target tissues, compared to the traditional methods. In this study, a novel method of NPs preparation was developed. The trimethyl chitosan (TMC) was synthesized using low molecular weight chitosan and was used for the preparation of TMC-NPs through ionic gelation method. Afterward, AmB-loaded TMC-NPs (TMC-NPs/AmB) were prepared and their drug delivery potential was testes. The TMC-NPs and TMC-NPs/AmB were characterized for their structure, particle size, Zeta potential, polydispersity index, morphology, loading efficiency, loading capacity, in vitro release profile, release kinetic, and entrapped AmB potency. The cytotoxicity and antifungal activity of TMC-NPs/AmB against Candida albicans biofilm were evaluated. The quaternization of TMC was estimated to be 36.4%. The mean particle size of TMC-NPs and TMC NPs/AmB were 210±15 and 365±10 nm, respectively, with a PDI of 0.30 and 0.4, ZP of +34±0.5 and +28±0.5 mV, respectively. Electron microscopy analysis indicated uniform spherical shapes with smooth surfaces. The TMC-NPs/AmB indicated LE of 76% and LC of 74.04 % with a potency of 110%. The release profile of TMC-NPs/AmB was best explained by the Higuchi model. The initial release after 10 h was obtained at 38%, and the rates of release after 36 and 84 h were determined at 67% and 76% respectively, which was significantly different (P<0.05) from previous time points. The minimum inhibitory concentration (MIC) (50%) of NPs/AmB and AmB were 0.65 and 1.75 μg/mL, and the MIC 80% were determined at 1.95 and 7.75 μg/mL, respectively, demonstrating a significant improvement in antifungal activity. The half-maximal inhibitory concentration for TMC-NPs/AmB and AmB were estimated at 86 and 105 μg/mL, respectively, indicating a significant reduction in cytotoxicity and the adverse effect. This study could successfully introduce a practical method to synthesize TMC-NPs. The encapsulation process was efficient and significantly improved the antifungal activity of AmB. The developed method can be applied to improve the feasibility of oral delivery while reducing the adverse effects associated with traditional methods.


Main Subjects

Article Title [French]

Un Nouveau Amphotéricine B Chargé de Nanoparticules de Triméthyl Chitosan Comme Système D'administration de Médicaments et Activité Antifongique sur le Biofilm de Candida albicans

Abstract [French]

L'amphotéricine B (AmB) est un agent antifongique efficace. Cependant, l'application d'AmB est associée à un certain nombre d'inconvénients et d'effets secondaires. L'application de nanoparticules (NPs) est connue pour permettre une administration de médicament plus efficace aux tissus cibles que les méthodes traditionnelles. Dans cette étude, le triméthyl chitosan (TMC) a été synthétisé à partir de chitosan de bas poids moléculaire. Le TMC a ensuite été utilisé pour la préparation de TMC-NPs par la méthode de gélification ionique. Ensuite, des TMC-NPs chargés en AmB (TMC-NPs/AmB) ont été préparés et leur application potentielle pour l'administration de médicaments. Les TMC-NPs et TMC-NPs /AmB ont été caractérisés pour leur structure, la taille des particules, le potentiel zêta (PZ), l'indice de polydispersité (IPD), la morphologie, l'efficacité de chargement (EC), la capacité de charge (LC), le profil de libération in vitro, libérer la puissance cinétique et piégée d'AmB. Ensuite, la cytotoxicité et l'activité antifongique des TMC-NPs/ AmB contre le biofilm de Candida albicans ont été évaluées. Dans cette étude, le pourcentage de quaternisation de TMC est estimé à 36.4. La taille moyenne des particules de TMC-NPs et TMC NPs/AmB était respectivement de 210±15 et 365±10 nm, avec un IPD de 0.30 et 0.40, ZP de +34± 0.5 et +28±0.5 mV respectivement. L'analyse au microscope électronique a indiqué des formes sphériques uniformes avec des surfaces lisses. Le TMC-NPs/AmB a indiqué EC de 76% et LC de 74.04% avec une puissance de 110%. Le profil de libération des TMC-NPs/AmB a été mieux expliqué par le modèle Higuchi. La libération initiale après 10 h était de 38% et le taux de libération après 36 et 84 h était de 67% et 76% respectivement, ce qui était significativement différent (P<0.05) des points de temps précédents. La concentration minimale inhibitrice (CMI) à 50% des NPs/AmB et AmB était de 0.65 et 1.75 μg/mL et la CMI de 80% était de 1.95 et 7.75 μg/mL, démontrant respectivement une amélioration significative de l'activité antifongique. La demi-concentration inhibitrice maximale pour les TMC-NPs/AmB et AmB était de 86 et 105 μg/mL respectivement, indiquant une réduction significative de la cytotoxicité et de l'effet indésirable. Cette étude pourrait introduire avec succès une méthode pratique pour synthétiser un TMC-NPs. Le processus d'encapsulation a été efficace et a considérablement amélioré l'activité antifongique de l'AmB. La méthode développée peut être appliquée pour améliorer la faisabilité de l'administration orale tout en réduisant les effets indésirables associés aux méthodes traditionnelles.

Keywords [French]

  • Amphotéricine B
  • Nanoparticules
  • Triméthyl chitosane
  • Candida albicans
  • biofilm
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