CHITOSAN NANOPARTICLES AS DRUG DELIVERY SYSTEM FOR CEPHALEXIN AND ITS ANTIMICROBIAL ACTIVITY AGAINST MULTIIDRUG RESISTENT BACTERIA

Authors

  • MONA IBRAHIM EL-ASSAL Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmaceutical Sciences and Pharmaceutical Industries, Future University in Egypt, 11835, Cairo, Egypt
  • NAGWAN GALAL EL-MENOFY Department of Microbiology and Immunology, Faculty of Pharmacy-Girls, Al-Azhar University, 11884, Cairo, Egypt

DOI:

https://doi.org/10.22159/ijpps.2019v11i7.33375

Keywords:

Antimicrobial activity, Antibiofilm, Cephalexin, Chitosan, Cytotoxicity assay, Drug delivery, Nanoparticles

Abstract

Objective: The evolution of antimicrobial resistance is a universal obstacle that necessities the innovation of more effective and safe antimicrobial alternatives with synergistic properties. The purpose of this study was to investigate the possible improvement of cephalexin antimicrobial treatments by loading into chitosan-based nanoparticles, then evaluate their antibacterial and antibiofilm activities as well as determination of its cytotoxicity.

Methods: Chitosan nanoparticles (CSNPs) were prepared by ionic gelation method. Parameters were studied to optimize the particle size of CSNPs including pH, stirring rate, homogenization and ultra-sonication time. Size was measured by transmission electron microscope (TEM) and Zeta sizer, morphology seen by scanning electron microscope (SEM). Entrapment efficiency, drug loading and drug content were calculated. Stability of both plain and loaded chitosan Nano-carriers, Drug release and Kinetics also compatibilities were studied. Antimicrobial activity of CSNPs and cephalexin loaded CSNPs were evaluated against 4 Gram-positive and 4 Gram-negative standard and clinical isolates by microdilution method, also assessment of antibiofilm activity of both formulas was investigated against two biofilm producers clinical isolates by tube assay in addition to determination of their cytotoxicity by MTT(3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

Results: Chitosan nanoparticles and its loaded antibiotics proved compatible combination with small Zeta size, suitable Zeta potential, maximum EE% and drug-loading capacity, sustained controlled release properties followed diffusion kinetic model and six month stability studies. Cephalexin loaded CSNPs showed better antimicrobial activity than plain CSNPs. Synergistic effects were found against S. aureus (ATCC 25923), B. subtilis (ATCC 9372), S. epidermidis, E. faecalis, P. aeruginosa (ATCC 29853) in addition to two carbapenem resistant isolates k. pneumoniae and E. coli. Also cephalexin loaded CSNPs exhibited antibiofilm activity against E. faecalis clinical isolate. Even though, cephalexin loaded CSNPs exhibited significant antibacterial activity, it showed less toxicity against mammalian cells, it had IC50 equal to 231.893 and did not exhibit any cytotoxicity against the WI-38 fibroblast cells at concentration 23.4 µg/ml.

Conclusion: Cephalexin loaded CSNPs possessed good stability and sustained release effect in addition to its antimicrobial, antibiofilm activities and reduced cytotoxicity.

Downloads

Download data is not yet available.

References

Raffi, Muhammad. Investigations into the antibacterial behavior of copper nanoparticles against Escherichia coli. Ann Microbiol 2010;60(Suppl 1):75-80.‏

Zaki NM, Hafez MM. Enhanced antibacterial effect of ceftriaxone sodium-loaded chitosan nanoparticles against intracellular Salmonella typhimurium. AAPS PharmSciTech 2012;13(Suppl 2):411-21.

Chattopadhyay D, Inamdar M. Improvement in properties of cotton fabric through synthesized nano-chitosan application. Indian J Fibre Textil Res 2013;38:14-21.

Benhabiles M, Salah R, Lounici H, Drouiche N, Goosen M, Mameri N. Antibacterial activity of chitin, chitosan and its oligomers prepared from shrimp shell waste. Food Hydrocolloids 2012;29(Suppl 1):48-56.

Cruz Romero M, Murphy T, Morris M, Cummins E, Kerry J. Antimicrobial activity of chitosan, organic acids and nano-sized solubilisates for potential use in smart antimicrobially-active packaging for potential food applications. Food Control 2013;34(Suppl 2):393-7.

Nagy A, Harrison A, Sabbani S, Munson JrRS, Dutta PK, Waldman WJ. Silver nanoparticles embedded in zeolite membranes: release of silver ions and mechanism of antibacterial action. Int J Nanomed 2011;6:1833–52.

Cuero R, Osuji G, Washington A. Carboxymethyl chitosan inhibition of aflatoxin production: role of zinc. Biotechnol Lett 1991;13:441–4.

Jimtaisong A, Saewan N. Utilization of carboxymethyl chitosan in cosmetics. Int J Cosmetic Sci 2014;36(Suppl 1):12-21.

El-Sherbiny IM, El-Baz NM. A review on bio nanocomposites based on chitosan and its derivatives for biomedical applications, eco-friendly polymer nanocomposites. Chem Appl 2015;74-173. Doi: 10.1007/978-81-322-2473-0_6

GOY Rejane C, BRITTO Douglas de ASSIS, Odilio BG. A review of the antimicrobial activity of chitosan. Polimeros 2009;19(Suppl 3):241-7.

Jia Z, Xu W. Synthesis and antibacterial activities of quaternary ammonium salt of chitosan. Carbohydr Res 2001;333(Suppl 1):1-6.

Lu Y, Cheng D, Lu S, Huang F, Li G. Preparation of quaternary ammonium salt of chitosan nanoparticles and their textile properties on antheraea pernyi silk modification. Textile Res J 2014;84(Suppl 19):2115-24.

Chung YC, Su YP, Chen CC, Jia G, Wang HL, Wu JCG, et al. Relationship between antibacterial activity of chitosan and surface characteristics of cell wall. Acta Pharmacol Sin 2004;25(Suppl 7):932-6.

Van der Lubben I, Verhoef J, Borchard G, Junginger H. Chitosan for mucosal vaccination. Adv Drug Delivery Rev 2001;52(Suppl 2):139-44.

Sawaengsak C, Mori Y, Yamanishi K, Mitrevej A, Sinchaipanid N. Chitosan nanoparticle encapsulated hemagglutinin-split influenza virus mucosal vaccine. AAPS PharmSciTech 2014;15(Suppl 2):317-25.

Del Guidice G, Baudner B. Mucosal vaccines with chitosan adjuvant and meningococcal antigens; 2015.

Avadi M, Sadeghi A, Tahzibi A, Bayati K, Pouladzadeh M, Zohuriaan-Mehr M, et al. Diethylmethyl chitosan as an antimicrobial agent: Synthesis, characterization and antibacterial effects. Eur Polym J 2004;40(Suppl 7):1355-61.

Abdel Fattah WI, Sallam ASM, Attawa N, Salama E, Maghraby AM, Ali GW. Functionality, antibacterial efficiency and biocompatibility of nanosilver/chitosan/silk/phosphate scaffolds 1. Synthesis and optimization of nanosilver/chitosan matrices through gamma rays irradiation and their antibacterial activity. Mater Res Express 2012;1(Suppl 3):35-24.

Garrait G, Beyssac E, Subirade M. Development of a novel drug delivery system: chitosan nanoparticles entrapped in alginate microparticles. J Microencapsulation 2014;31(Suppl 4):363-72.

TiyaboonchaI W. Chitosan nanoparticles: a promising system for drug delivery. Naresuan University J 2003;11(Suppl3):51-66.

Kim TH, Park IK, Nah JW, Choi YJ, Cho CS. Galactosylated chitosan/DNA nanoparticles prepared using water-soluble chitosan as a gene carrier. Biomaterials 2004;25(Suppl 17):3783-92.

Chaisri W, Hennink WE, Okonogi S. Preparation and characterization of cephalexin loaded PLGA microspheres. Curr Drug Delivery 2009;6:69-75.

Speight TM, Brogden RN, Avery GS. Cephalexin: a review of its antibacterial, pharmacological and therapeutic properties. Drugs 1972;3:9-78.‏

El Menofy NG. Phenotypic and molecular detection of new delhi metallo-betalactamase-1 (bla NDM-1) gene among Escherichia coli and Klebsiella pneumoniae in hospitalized Egyptian patients. New Egypt J Microbiol 2018;51:1-14.

El-Assal MIA. Proniosomes as nano-carrier for transdermal delivery of atenolol niosomal gel. Int J Drug Delivery Technol 2017;7:283-97.

Yasmin MB, Prathyusha RG. Formulation and evaluation of dasatinib loaded solid lipid nanoparticles. Int J Pharm Pharm Sci 2018;10:14-20.

Hu CJ, Rhodes DG. Proniosomes: a novel drug carrier preparation. Int J Pharm 1999;185:23–35.

Yadav SK. Nanoscale materials in targeted drug delivery. Theragnosis and Tissue Regeneration. New York: Springer; 2016.

Al Kassas RS, Al Gohary OM, Al Faadhel MM. Controlling of systemic absorption of gliclazide through incorporation into alginate beads. Int J Pharm 2007;341:230-7.

Shweta P. Formulation and evaluation of garlic powder loaded floating matrix tablet. Int J Pharm Pharm Sci 2019;11:17-21.

Shu XZ, Zhu KJ, Song W. Controlled drug release properties of ionically cross-linked chitosan beads: the influence of anion structure. Int J Pharm 2002;233:217-25.

Pourjavadi A, Barzegar Sh, GR Mahdavinia. MBA-crosslinked Na-Alg/CMC as a smart full-polysaccharide superabsorbent hydrogels. Carbohydr Polym 2006;66:386–95.

Pardakhty A, Varshosaz J, Rouholamini A. In vitro study of polyoxyethylene alkyl ether niosomes for delivery of insulin. Int J Pharm 2007;328:130–41.

Novy P, Davidova H, Serrano Rojero CS, Rondevaldova J, Pulkrabek Kokoska. Composition and antimicrobial activity of Euphrasia rostkoviana Hayne essential oil. J Evidence Based Complementary Altern Med 2015:5. http://dx.doi.org/10.1155/2015/734101

Jamaran S, Zarif BR. Synergistic effect of silver nanoparticles with neomycin or gentamicin antibiotics on mastitis-causing Staphylococcus aureus. Open J Ecol 2016;6(Suppl 7):452.

Fratini F. A novel interpretation of the fractional inhibitory concentration index: the case origanum vulgare L. and leptospermum scoparium JR et G. Forst essential oils against Staphylococcus aureus strains. Microbiol Res 2017;195:11-7.

Hasan Y, Ozeki, SR Kabir. Purification of a novel chitin-binding lectin with antimicrobial and antibiofilm activities from a Bangladeshi cultivar of potato (Solanum tuberosum). Indian J Biochem Biophysics 2014;51(Suppl 2):142-8.

Hussein Al Ali, Samer Hasan. Preparation of chitosan nanoparticles as a drug delivery system for perindopril erbumine. Polymer Composites 2018;39(Suppl 2): 544-52.‏

Kittur FS, Harish Prashanth KV, Udaya Sankar K, Tharanathan RN. Characterization of chitin, chitosan and their carboxymethyl derivatives by differential scanning calorimetry. Carbohydrate Polymers 2002;49:185-93.

Di Stefano R, Scopelliti M, Pellerito C, Casella G, Fiore T, Stocco GC, et al. Organometallic complexes with biological molecules. XVIII. Alkyltin (IV) cephalexinate complexes: synthesis, solid state and solution phase investigations. J Inorg Biochem 2004;98(Suppl 3):534-46.

Xu WZ, Wang SQ, Li AJ, Wang XL. Synthesis of aminopropyltriethoxysilane grafted/tripolyphosphate intercalated ZnAl LDHs and their performance in the flame retardancy and smoke suppression of polyurethane elastomer. RSC Adv 2016;6:48189–98.

Xu Y, Du Y. Effect of molecular structure of chitosan on protein delivery properties of chitosan nanoparticles. Int J Pharm 2003;250:215-26.

Knaul JZ, Hudson SM, Creber KAM. Improved mechanical properties of chitosan fibres. J Appl Polym Sci 1999;72:1721-31.

Wang X, Ma J, Wang Y, He B. Structural characterization of phosphorylated chitosan and their applications as effective additives of calcium phosphate cements. Biomaterials 2001;22:2247-55.

Noyes A, Itney W. The rate of solution of solid substances in their own solutions. J Am Chem Soc 1897;19:934.

Higuchi T. Mechanism of sustained–action medication theoretical analysis of release of solid drugs dispersed in solid matrices. J Pharm Sci 1963;52:1145-9.

Macheras P, Dokoumetzidis A. On the heterogeneity of drug dissolution and release. Pharm Res 2000;17:108-12.

Friedman M, Juneja VK. Review of antimicrobial and antioxidative activities of chitosans in food. J Food Prot 2010;73(Suppl 9):1737-61.

Ghadi A, Mahjoub S, Tabandeh F, Talebnia F. Synthesis and optimization of Chitosan nanoparticles: potential applications in nanomedicine and biomedical engineering. Iran Caspian: J Intern Med 2014;5(Suppl 3):156-61.

Saini R, Saini S, Sharma S. Nanotechnology: the future medicine. J Cutan Aesthet Surg 2010;3(Suppl 1):32-3.

Wardani Giftania. In vitro antibacterial activity of chitosan nanoparticles against mycobacterium tuberculosis. Pharmacogn J 2018;10(Suppl 1):162-6.

Saha P, Goyal AK, Rath G. Formulation and evaluation of chitosan-based ampicillin trihydrate nanoparticles. Trop J Pharm Res 2010;9:483-8.‏

Xu J, Xu B, Shou D, Xia X, Hu Y. Preparation and evaluation of vancomycin-loaded N-trimethyl chitosan nanoparticles. Polymers 2015;7(Suppl 9):1850-70.

Ibrahim HM, El-Bisi MK, Taha GM, El-Alfy EA. Chitosan nanoparticles loaded antibiotics as drug delivery biomaterial. J Appl Pharm Sci 2015;5(Suppl 10):85-90.

Thaya Rajagopalan. Synthesis of chitosan-alginate microspheres with high antimicrobial and antibiofilm activity against multi-drug resistant microbial pathogens. Microbial Pathogenesis 2018;114:17-24.

Kanchana M, Malathy BR, Jeevitha D, Reema O, Sonia S. Synthesis and evaluation of amoxicillin and cephalexin encapsulated chitosan nanoparticles against urinary tract infection causing Escherichia coli and Klebsiella pneumoniae. World J Pharm Res 2014;3(Suppl 2):512-26.

Divya K, Vijayan S, George TK, Jisha MS. Antimicrobial properties of chitosan nanoparticles: mode of action and factors affecting activity. Fibers Polymers 2017;18(Suppl 2):221-30.

Tamara F, Lin C, Mi FL, Ho YC. Antibacterial effects of chitosan/cationic peptide nanoparticles. Nanomaterials 2018;8(Suppl 2):88.

Li J, McL, Sborough L. The effects of the surface charge and hydrophobicity of Escherichia coli on its adhesion to beef muscle. Int J Food Microbiol 1999;53:185-93.

Rabea EI, Badawy ME, Stevens CV, Smagghe G, Steurbaut W. Chitosanas antimicrobial agent: applications and mode of action. Bio Macromolecules 2003;4:1457–65.

Divya K, Jisha MS. Chitosan nanoparticles preparation and applications. Environ Chem Lett 2018;16:101–12.

Zheng LY, Zhu JF. Study on antimicrobial activity of chitosan with different molecular weights. Carbohydr Polym 2003;54:527–630.

Goy RC, Morais ST, Assis OB. Evaluation of the antimicrobial activity of chitosan and its quaternized derivative on E. coli and S. aureus growth. Rev Bras Farmacogn 2016;26(Suppl 1):122-7.‏

Ramachandran R, Sangeetha D. Antibiofilm efficacy of silver nanoparticles against biofilm forming multidrug resistant clinical isolates. Pharma Innovation 2017;6(11, Part A):36.

Hassan A, Usman J, Kaleem F, Omair M, Khalid A, Iqbal M. Evaluation of different detection methods of biofilm formation in the clinical isolates. Braz J Infect Dis 2011;15(Suppl 4):305-11.

Costa EM, Silva S, Pina C, Tavaria FK, Pintado M. Antimicrobial effect of chitosan against periodontal pathogens biofilms. SOJ Microbiol Infect Dis 2014;2(Suppl 1):1-6.‏

Namasivayam SKR, Roy EA. Enhanced antibiofilm activity of chitosan stabilized chemogenic silver nanoparticles against Escherichia coli. Int J Sci Res Publications 2013;3(Suppl 4):1-9.‏

Kalishwaralal K, Barath Mani Kanth S, Pandian SRK, Deepak V, Gurunathan S. Silver nanoparticles impede the biofilm formation by Pseudomonas aeruginosa and Staphylococcus epidermidis: Colloids Surf B 2010;79(Suppl 2):340-4.

Yeon SJ, Lee JW, Lee JW, Kwark YJ, Kim SH, Lee KY. Effect of nano-structured polymer surfaces on osteoblast adhesion and proliferation. J Controlled Release 2011;152:e257–58.

Salarian AA, Mollamahale YB, Hami Z, Soltani Rezaee Rad M. Cephalexin nanoparticles: synthesis, cytotoxicity and their synergistic antibacterial study in combination with silver nanoparticles. Materials Chem Physics 2017;198:125-30.‏

Published

01-07-2019

How to Cite

EL-ASSAL, M. I., and N. G. EL-MENOFY. “CHITOSAN NANOPARTICLES AS DRUG DELIVERY SYSTEM FOR CEPHALEXIN AND ITS ANTIMICROBIAL ACTIVITY AGAINST MULTIIDRUG RESISTENT BACTERIA”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 11, no. 7, July 2019, pp. 14-27, doi:10.22159/ijpps.2019v11i7.33375.

Issue

Section

Original Article(s)