Optimizing indomethacin-loaded chitosan nanoparticle size, encapsulation, and release using Box–Behnken experimental design
Introduction
Chitosan is a cationic hydrophilic linear polysaccharide macromolecule of biological origin. This biocompatible/environment-friendly compound is composed of deacetylated unit (β-(1–4)-linked d-glucosamine) and acetylated unit (N-acetyl-d-glucosamine). Chitosan has mucoadhesive and membrane-permeability enhancing properties [1]. It has numerous advantages for mucosal delivery such as biodegradability and low toxicity [2]. It is being used as an excipient in various formulations including micro- and nanoparticles [3]. Multiple methods were reported to prepare chitosan nanoparticles including self-assembly technique through chemical modification, complex coacervation process, emulsion-droplet coalescence, ionotropic gelation and other techniques. Among these techniques, ionotropic gelation [4] is preferred for its relative simplicity, convenience, and the rid of high temperature and organic solvents; hence, sufficient encapsulation of therapeutic agents such as doxorubicin [5], cyclosporine-A [6] as well as proteins [7] could be possible.
Phytochemicals like catechins have been fabricated and encapsulated in chitosan-TPP nanoparticles with up to 53% encapsulation efficiency [8]. Chen and Subirade developed chitosan and TPP based nanoparticles to encapsulate a vitamin riboflavin [9]. Also, Desai et al. developed sustained release microspheres of cross linked chitosan loaded with vitamin-C by spray drying [10]. Moreover, the coating of alginate beads with chitosan to encapsulate the living microbial supplements (probiotics) was developed by Le-Tien et al. [11]. During encapsulation of Lactobacillus acidophilus and Lactobacillus casei in microspheres, chitosan coated alginate beads provided better protection for these two probiotics as compared to poly-L-lysine-coated alginate beads [12]. Similarly, L. acidophilus-547 and L. casei-01 strains were best protected by chitosan-coated alginate beads [13]. Unsaturated fatty acids functions as neuroprotective, antioxidant, and anti-inflammatory, but they are highly susceptible to oxidative rancidity; therefore, chitosan could be used to stabilized the o/w-emulsions by getting adsorbed on to the oil droplets and form a protective layer by reducing the interfacial tension. Spray dried tuna o/w-emulsion was stabilized by chitosan-lecithin and found oxidative stable as compared to buck oils, hence were found an excellent ω-3 fatty acid components for the functional foods [14]. Immobilization carriers for enzymes were possible by formulating the chitosan based macro-, micro- and nano-sized particles by precipitation, emulsification, and ionotropic gelation methods, respectively. For example, the highest activity and excellent storage stability of β-galactosidase was found when they were immobilized on chitosan-nanoparticles prepared by ionotropic gelation technique, where sodium sulfate was the gelation agent [15]. Flavors like citral and limonene in emulsion forms were stabilized by sodium dodecyl sulfate-chitosan and were found more effective at hindering the citral oxidation product formation than gum Arabica- stabilized emulsions. Similarly, emulsion of limonene was stabilized by sodium dodecyl sulfate-chitosan, and formation of limonene oxide and carvone were found very low as compared to gum Arabica-stabilized emulsions at pH 3.0. The sodium dodecyl sulfate-chitosan multilayer has ability to form a thick cationic emulsion droplet interface that inhibits the oxidative deterioration of citral and limonene [16].
In this study, we applied ionotropic gelation technique for the preparation of indomethacin-loaded chitosan nanoparticles. Due to the formation of inter- and intramolecular cross-linkages, chitosan gelation occurs when it comes in contact with certain polyanions [5]. In our study, tripolyphosphate was used a cross linker. Although ionotropic gelation of chitosan with tripolyphosphate was firstly reported by Bodmeier et al. [17], our main intention is to apply Box–Behnken design for the optimization of chitosan nanoparticles while assessing the effects of different preparation parameters on their physicochemical characteristics viz. particle size, encapsulation and release drug. In this study indomethacin was used as a model drug, however the developed nanoparticles can be used, in principle, to encapsulate hydrophilic or hydrophobic therapeutic agents, bacteria, food, or biochemical ingredients.
Section snippets
Materials
Indomethacin (Lot No. 134737/42) was purchased from its manufacturer Winlab, UK. High purity, molecular weight 140K-220K, deacetylated chitin (chitosan) and the cross linker tripolyphosphate (TPP) were purchased from Sigma–Aldrich (St. Louis, MO, USA). Acetic acid glacial was purchased from BDH Limited (Poole, England). Dichloromethane and acetonitrile (HiPerSolv CHROMANORM for HPLC grade) were purchased from BDH, PROLABO®, LEUVEN, EC. Purified water was obtained by Milli-Q® water purifier
Results and discussions
In the present study we have chosen a lipophilic drug, indomethacin, to encapsulate into chitosan-TPP nanoparticles as a model drug, and evaluated the morphology of the produced nanoparticles by transmission electron microscopy (TEM), particle size, polydispersity, zeta potential, encapsulation efficiency, drug loading, swelling behavior of chitosan nanoparticles in water, and in-vitro drug release. Our results have given indications that the developed nanoparticles by ionic gelation method
Conclusions
Using ionotropic gelation method, good average sized positively charged chitosan-nanoparticles with high encapsulation efficiency; satisfactory drug release and good storage stability properties could be produced and optimized by using a three-factor, three-level Box–Behnken design. High magnitudes of zeta-potential of the NPs indicated a stable dispersive nature of the developed chitosan-NPs.The quantitative effect of the chosen independent factors at different levels to achieve minimum
Conflict of interest
The authors declare no competing financial interest.
Acknowledgment
This project was funded by the Research Groups Program (Research Group number RG-1436-027), Deanship of Scientific Research, King Saud University, Riyadh, Saudi Arabia.
References (44)
- et al.
Chitosan nanoparticles loaded with dorzolamide and pramipexole
Carbohydr. Polym.
(2008) - et al.
Chitosan nanoparticles: preparation, size evolution and stability
Int. J. Pharm.
(2013) - et al.
Chitosan—a versatile semi-synthetic polymer in biomedical applications
Prog. Polym. Sci.
(2011) - et al.
Chitosan nanoparticles as delivery systems for doxorubicin
J. Control Release
(2001) - et al.
Chitosan nanoparticles: a new vehicle for the improvement of the delivery of drugs to the ocular surface. Application to cyclosporin A
Int. J. Pharm.
(2001) - et al.
Microencapsulated chitosan nanoparticles for pulmonary protein delivery: in vivo evaluation of insulin-loaded formulations
J. Control Release
(2012) - et al.
Chitosan/beta-lactoglobulin core-shell nanoparticles as nutraceutical carriers
Biomaterials
(2005) - et al.
The influence of coating materials on some properties of alginate beads and survivability of microencapsulated probiotic bacteria
Int. Dairy J.
(2004) - et al.
Survival of probiotics encapsulated in chitosan-coated alginate beads in yoghurt from UHT- and conventionally treated milk during storage
LWT-Food Sci. Technol.
(2006) - et al.
Preparation of chitosan particles suitable for enzyme immobilization
J. Biochem. Biophys. Methods
(2008)
Chitosan-sodium alginate nanoparticles as submicroscopic reservoirs for ocular delivery: formulation, optimisation and in vitro characterisation
Eur. J. Pharm. Biopharm.
Development and bioavailability assessment of ramipril nanoemulsion formulation
Eur. J. Pharm. Biopharm.
Synthesis, characterization and in vitro drug release of magnetic N-benzyl-O-carboxymethylchitosan nanoparticles loaded with indomethacin
Acta Biomater.
Development and validation of HPLC method for determination of indomethacin and its two degradation products in topical gel
J. Pharm. Biomed. Anal.
Solid lipid nanoparticles loaded with insulin by sodium cholate-phosphatidylcholine-based mixed micelles: preparation and characterization
Int. J. Pharm.
Synthesis and characterisation of chitosan crosslinked-β-cyclodextrin grafted silylated magnetic nanoparticles for controlled release of Indomethacin
J. Magn. Magn. Mater.
Swelling and drug release properties of acrylamide/carboxymethyl cellulose networks formed by gamma irradiation
Radiat. Phys. Chem.
Sigmoidal release of indomethacin from pectin matrix tablets: effect of in situ crosslinking by calcium cations
Int. J. Pharm.
Understanding the quality of protein loaded PLGA nanoparticles variability by Plackett–Burman design
Int. J. Pharm.
Effects of molecular weight, degree of acetylation and ionic strength on surface tension of chitosan in dilute solution
Carbohydr. Polym.
Formation mechanism of monodisperse, low molecular weight chitosan nanoparticles by ionic gelation technique
Colloids Surf. B Biointerfaces
Structure and interactions in covalently and ionically crosslinked chitosan hydrogels for biomedical applications
Eur. J. Pharm. Biopharm.
Cited by (66)
Metronidazole loaded chitosan–phytic acid polyelectrolyte complex nanoparticles as mucoadhesive vaginal delivery system for bacterial vaginosis
2024, International Journal of Biological MacromoleculesDevelopment and optimization of in-situ gel containing chitosan nanoparticles for possible nose-to-brain delivery of vinpocetine
2023, International Journal of Biological MacromoleculesOral docetaxel delivery with cationic polymeric core-shell nanocapsules: In vitro and in vivo evaluation
2023, Journal of Drug Delivery Science and TechnologyOptimization and Appraisal of Chitosan-Grafted PLGA Nanoparticles for Boosting Pharmacokinetic and Pharmacodynamic Effect of Duloxetine HCl Using Box-Benkhen Design
2023, Journal of Pharmaceutical SciencesCitation Excerpt :Nevertheless, relating to the nonmucoadhesive characteristics of PLGA, there is an additional need to use mucoadhesives that can help nanoformulation's adhesion to the nasal mucosal surface and overwhelm nasal mucociliary clearance.12 Since the nasal mucosa is negatively charged, a naturally occurring positively charged biodegradable polymeric material, chitosan (CS), has been used as a mucoadhesive and penetration enhancing agent for stronger bond formation with the nasal epithelium and longer preparation retention time.13,14 CS as a natural biodegradable polymer, was subjected to enzymatic transformation into basic, nontoxic components.