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

Document Type : Original Articles

Authors

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

Abstract

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.

Keywords

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

References

  1. Sanchez DA, Schairer D, Tuckman-Vernon C, Chouake J, Kutner A, Makdisi J, et al. Amphotericin B releasing nanoparticle topical treatment of Candida spp. in the setting of a burn wound. Nanomed-Nanotechnol. 2014;10(1):269-77.
  2. Grela E, Zdybicka-Barabas A, Pawlikowska-Pawlega B, Cytrynska M, Wlodarczyk M, Grudzinski W, et al. Modes of the antibiotic activity of amphotericin B against Candida albicans. Sci Rep. 2019;9(1):1-10.
  3. Roy G, Galigama RD, Thorat VS, Mallela LS, Roy S, Garg P, et al. Amphotericin B containing microneedle ocular patch for effective treatment of fungal keratitis. Int J Pharm. 2019;572:118808.
  4. Dominguez E, Zarnowski R, Choy H, Zhao M, Sanchez H, Nett J, et al. Conserved Role for Biofilm Matrix Polysaccharides in Candida auris Drug Resistance. mSphere. 2019;4.
  5. Dominguez E, Zarnowski R, Choy HL, Zhao M, Sanchez H, Nett J, et al. Conserved Role for Biofilm Matrix Polysaccharides in Candida auris Drug Resistance. mSphere. 2019;4(1):e00680-18.
  6. Wang JJ, Zeng ZW, Xiao RZ, Xie T, Zhou GL, Zhan XR, et al. Recent advances of chitosan nanoparticles as drug carriers. Int J Nanomedicine. 2011;6:765-74.
  7. Singh G, Kaur T, Kaur A, Kaur R, Kaur R. Analytical methods for determination of Amphotericin B in biological samples: a short review. JABS. 2014;1:R26-R32.
  8. Salah R, editor Antileishmanial activities of chitin and chitosan prepared from shrimp shell waste. Int Conf Chem Mettalurgy Environ Eng; 2015.
  9. Vongchan P, Wutti-In Y, Sajomsang W, Gonil P, Kothan S, Linhardt RJ. N, N, N-Trimethyl chitosan nanoparticles for the delivery of monoclonal antibodies against hepatocellular carcinoma cells. Clin Microbiol  Infect. 2011;85(1):215-20.
  10. Wu J, Huang Y, Yao R, Deng S, Li F, Bian X. Preparation and characterization of starch nanoparticles from potato starch by combined solid‐state acid‐catalyzed hydrolysis and nanoprecipitation. Starch Starke. 2019;71(9-10):1900095.
  11. Cakic M, Glisic S, Nikolic G, Nikolic GM, Cakic K, Cvetinov M. Synthesis, characterization and antimicrobial activity of dextran sulphate stabilized silver nanoparticles. J Mol. 2016;1110:156-61.
  12. Xu C, Rezeng C, Li J, Zhang L, Yan Y, Gao J, et al. 1H NMR-Based Metabolomics Study of the Toxicological Effects in Rats Induced by “Renqing Mangjue” Pill, a Traditional Tibetan Medicine. Front Pharmacol. 2017;8:602.
  13. Perez AP, Altube MJ, Schilrreff P, Apezteguia G, Celes FS, Zacchino S, et al. Topical amphotericin B in ultradeformable liposomes: Formulation, skin penetration study, antifungal and antileishmanial activity in vitro. Colloids Surf B. 2016;139:190-8.
  14. Farhadian A, Dounighi NM, Avadi M. Enteric trimethyl chitosan nanoparticles containing hepatitis B surface antigen for oral delivery. Hum Vaccin Immunother. 2015;11(12):2811-8.
  15. Wu M, Long Z, Xiao H, Dong C. Recent research progress on preparation and application of N, N, N-trimethyl chitosan. Carbohydr Res. 2016;434:27-32.
  1. Ofori-Kwakye K, Mfoafo KA, Kipo SL, Kuntworbe N, El Boakye-Gyasi M. Development and evaluation of natural gum-based extended release matrix tablets of two model drugs of different water solubilities by direct compression. Saudi Pharm J. 2016;24(1):82-91.
  2. Xu J, Xu B, Shou D, Xia X, Hu Y. Preparation and evaluation of vancomycin-loaded N-trimethyl chitosan nanoparticles. Polymers. 2015;7(9):1850-70.
  3. AL-Quadeib BT, Radwan MA, Siller L, Horrocks B, Wright MC. Stealth Amphotericin B nanoparticles for oral drug delivery: In vitro optimization. Saudi Pharm J. 2015;23(3):290-302.
  4. Zarifpour M, Hadizadeh F, Iman M, Tafaghodi M. Preparation and characterization of trimethyl chitosan nanospheres encapsulated with tetanus toxoid for nasal immunization studies. Pharm Sci. 2013;18(4):193-8.
  5. Yan C, Hu CQ. Improvement of microbiological potency measurement of amphotericin B. Chinese Antibiot. 2013;38:434-9.
  6. Pharmacopoeia U, editor National Formulary.< 81> Antibiotics-Microbial assays. USP32-NF27, US Pharmacopeial Convention Inc, Rockville, USA; 2009.
  7. De Jong WH, Borm PJ. Drug delivery and nanoparticles: applications and hazards. Int J Nanomed. 2008;3(2):133.
  8. Pant A, Negi JS. Novel controlled ionic gelation strategy for chitosan nanoparticles preparation using TPP-β-CD inclusion complex. Eur J Pharm SCI. 2018;112:180-5.
  9. Mourya VK, Inamdar NN. Trimethyl chitosan and its applications in drug delivery. J Mater Sci Mater Med. 2009;20(5):1057-79.
  10. Rebelatto MC, Guimond P, Bowersock TL, HogenEsch H. Induction of systemic and mucosal immune response in cattle by intranasal administration of pig serum albumin in alginate microparticles. Vet Immunol Immunopathol. 2001;83(1):93-105.
  11. Yien L, Zin NM, Sarwar A, Katas H. Antifungal activity of chitosan nanoparticles and correlation with their physical properties. Int J Biomater. 2012;2012.
  12. Chen F, Zhang Z-R, Huang Y. Evaluation and modification of N-trimethyl chitosan chloride nanoparticles as protein carriers. Int J Pharm. 2007;336(1):166-73.
Archives of Razi Institute
Volume 76, Issue 3
September 2021
Pages 575-590

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