Cationic Chitosan Derivatives Research Articles

Preparation and Characterization of Novel Cationic Chitosan Derivatives Bearing Quaternary Ammonium and Phosphonium Salts and Assessment of Their Antifungal Properties.

Chitosan is an abundant and renewable polysaccharide, its derivatives exhibit attractive bioactivities and the wide applications in various biomedical fields. In this paper, two novel cationic chitosan derivatives modified with quaternary phosphonium salts were successfully synthesized via trimethylation, chloride acetylation, and quaternization with tricyclohexylphosphine and triphenylphosphine. The structures and properties of synthesized products in the reactions were characterized by FTIR spectroscopy, 1H-NMR, 31P-NMR, elemental and thermogravimetric analysis. The antifungal activities of chitosan derivatives against four kinds of phytopathogens, including Phomopsis asparagi, Watermelon fusarium, Colletotrichum lagenarium, and Fusarium oxysporum were tested using the radial growth assay in vitro. The results revealed that the synthesized cationic chitosan derivatives showed significantly improved antifungal efficiency compared to chitosan. It was reasonably suggested that quaternary phosphonium groups enabled the obviously stronger antifungal activity of the synthesized chitosans. Especially, the triphenylphosphonium-functionalized chitosan derivative inhibited the growth of Phomopsis asparagi most effectively, with inhibitory indices of about 80% at 0.5 mg/mL. Moreover, the data demonstrated that the substituted groups with stronger electron-withdrawing ability relatively possessed greater antifungal activity. The results suggest the possibility that cationic chitosan derivatives bearing quaternary phosphonium salts could be effectively employed as novel antifungal biomaterials for application in the field of agriculture.

Efficient Percutaneous Delivery of the Antimelanogenic Agent Glabridin Using Cationic Amphiphilic Chitosan Micelles.

Partially myristoylated chitosan pyrrolidone carboxylate (PMCP) is a cationic amphiphilic chitosan derivative. Glabridin (Glab) from licorice root extracts is a hydrophobic antimelanogenic agent. Here we assessed the effects of cationic Glab-containing polymeric micelles derived from PMCP (Glab/PMCP-PM) on the ability of Glab to penetrate the skin and inhibit melanogenesis using a human skin model. The amount of Glab absorbed 24 h after the application of Glab/PMCP-PM was approximately four times higher than that of conventional oil-in-water micelles (control) prepared using Tween 60. Further, the release of IL-1α, a mediator of inflammation, was not detected. Treatment with Glab/PMCP-PM significantly increased the inhibition of melanogenesis compared with control. The inhibition of melanogenesis depends upon the enhanced ability of Glab to penetrate the skin, particularly the epidermis. Moreover, the inhibition of melanogenesis and the cationic potential of the Glab/PMCP-PM levels were increased by the cationic phospholipid copolymer. Therefore, Glab/PMCP-PM shows potential as an effective transdermal delivery system for treating skin hyperpigmentation.

Chitosan Derivatives Active against Multidrug-Resistant Bacteria and Pathogenic Fungi: In Vivo Evaluation as Topical Antimicrobials.

The continuous rise of antimicrobial resistance and the dearth of new antibiotics in the clinical pipeline raise an urgent call for the development of potent antimicrobial agents. Cationic chitosan derivatives, N-(2-hydroxypropyl)-3-trimethylammonium chitosan chlorides (HTCC), have been widely studied as potent antibacterial agents. However, their systemic structure-activity relationship, activity toward drug-resistant bacteria and fungi, and mode of action are very rare. Moreover, toxicity and efficacy of these polymers under in vivo conditions are yet to be established. Herein, we investigated antibacterial and antifungal efficacies of the HTCC polymers against multidrug resistant bacteria including clinical isolates and pathogenic fungi, studied their mechanism of action, and evaluated cytotoxic and antimicrobial activities in vitro and in vivo. The polymers were found to be active against both bacteria and fungi (MIC = 125-250 μg/mL) and displayed rapid microbicidal kinetics, killing pathogens within 60-120 min. Moreover, the polymers were shown to target both bacterial and fungal cell membrane leading to membrane disruption and found to be effective in hindering bacterial resistance development. Importantly, very low toxicity toward human erythrocytes (HC50 = >10000 μg/mL) and embryo kidney cells were observed for the cationic polymers in vitro. Further, no inflammation toward skin tissue was observed in vivo for the most active polymer even at 200 mg/kg when applied on the mice skin. In a murine model of superficial skin infection, the polymer showed significant reduction of methicillin-resistant Staphylococcus aureus (MRSA) burden (3.2 log MRSA reduction at 100 mg/kg) with no to minimal inflammation. Taken together, these selectively active polymers show promise to be used as potent antimicrobial agents in topical and other infections.

Regioselective Sequential Modification of Chitosan via Azide-Alkyne Click Reaction: Synthesis, Characterization, and Antimicrobial Activity of Chitosan Derivatives and Nanoparticles.

Recently, the attention of researchers has been drawn toward the synthesis of chitosan derivatives and their nanoparticles with enhanced antimicrobial activities. In this study, chitosan derivatives with different azides and alkyne groups were synthesized using click chemistry, and these were further transformed into nanoparticles by using the ionotropic gelation method. A series of chitosan derivatives was successfully synthesized by regioselective modification of chitosan via an azide-alkyne click reaction. The amino moieties of chitosan were protected during derivatization by pthaloylation and subsequently unblocked at the end to restore their functionality. Nanoparticles of synthesized derivatives were fabricated by ionic gelation to form complexes of polyanionic penta-sodium tripolyphosphate (TPP) and cationic chitosan derivatives. Particle size analysis showed that nanoparticle size ranged from 181.03 ± 12.73 nm to 236.50 ± 14.32 nm and had narrow polydispersity index and positive surface charge. The derivatives and corresponding nanoparticles were evaluated in vitro for antibacterial and antifungal activities against three gram-positive and gram-negative bacteria and three fungal strains, respectively. The minimum inhibitory concentration (MIC) of all derivatives ranged from 31.3 to 250 µg/mL for bacteria and 188 to1500 µg/mL for fungi and was lower than that of native chitosan. The nanoparticles with MIC ranging from 1.56 to 25 µg/mLfor bacteria and 94 to 750 µg/mL for fungi exhibited higher activity than the chitosan derivatives. Chitosan O-(1-methylbenzene) triazolyl carbamate and chitosan O-(1-methyl phenyl sulfide) triazolyl carbamate were the most active against the tested bacterial and fungal strains. The hemolytic assay on erythrocytes and cell viability test on two different cell lines (Chinese hamster lung fibroblast cells V79 and Human hepatic cell line WRL68) demonstrated the safety; suggesting that these derivatives could be used in future medical applications. Chitosan derivatives with triazole functionality, synthesized by Huisgen 1,3-dipolar cycloaddition, and their nanoparticles showed significant enhancement in antibacterial and antifungal activities in comparison to those associated with native, non-altered chitosan.

The effect of substituent, degree of acetylation and positioning of the cationic charge on the antibacterial activity of quaternary chitosan derivatives.

A series of water-soluble cationic chitosan derivatives were prepared by chemoselective functionalization at the amino group of five different parent chitosans having varying degrees of acetylation and molecular weight. The quaternary moieties were introduced at different alkyl spacer lengths from the polymer backbone (C-0, C-2 and C-6) with the aid of 3,6-di-O-tert-butyldimethylsilyl protection of the chitosan backbone, thus allowing full (100%) substitution of the free amino groups. All of the derivatives were characterized using 1H-NMR, 1H-1H COSY and FT-IR spectroscopy, while molecular weight was determined by GPC. Antibacterial activity was investigated against Gram positive S. aureus and Gram negative E. coli. The relationship between structure and activity/toxicity was defined, considering the effect of the cationic group’s structure and its distance from the polymer backbone, as well as the degree of acetylation within a molecular weight range of 7–23 kDa for the final compounds. The N,N,N-trimethyl chitosan with 100% quaternization showed the highest antibacterial activity with moderate cytotoxicity, while increasing the spacer length reduced the activity. Trimethylammoniumyl quaternary ammonium moieties contributed more to activity than 1-pyridiniumyl moieties. In general, no trend in the antibacterial activity of the compounds with increasing molecular weight or degree of acetylation up to 34% was observed.

Chitosan‐Based Nanocoatings for Hypothermic Storage of Living Cells

The formation of ultrathin chitosan-based nanocoating on HL-60 model cells and their protective function in hypothermic storage are presented. HL-60 cells are encapsulated in ultrathin shells by adsorbing cationic and anionic chitosan derivatives in a stepwise, layer-by-layer, procedure carried out in an aqueous medium under mild conditions. The chitosan-based films are also deposited on model lipid bilayer and the interactions are studied using ellipsometry and atomic force microscopy. The cells covered with the chitosan-based films and stored at 4 °C for 24 h express viability comparable to that of the control sample incubated at 37 °C, while the unprotected cells stored under the same conditions do not show viability. It is shown that the chitosan-based shell protects HL-60 cells against damaging effect of hypothermic storage. Such nanocoatings provide protection, mechanical stability, and support the cell membrane, while ensuring penetration of small molecules such as nutrients/gases what is essential for cell viability.

Luminescent Transparent Composite Films Based on Lanthanopolyoxometalates and Filmogenic Polysaccharides

AbstractA series of polysaccharide‐based transparent composite films containing lanthanopolyoxometalates (LnPOMs) have been prepared by using a water‐soluble cationic chitosan derivative [HTCC = N‐(2‐hydroxypropyl)‐3‐trimethylammonium chitosan chloride] and pullulan (PUL). HTCC was more suitable than pure chitosan for this purpose due to its positive charge and solubility in water over a broad range of pH values. The LnPOMs [Ln(W5O18)2]9– (Ln = Eu and Tb) were incorporated into the polysaccharides. EuPOMs encapsulated in amorphous silica shells were also incorporated into the polysaccharides to test the effect on film processing of the loading of nanoparticles instead of LnPOM anions. The incorporation of LnPOMs or LnPOM/silica core/shell nanoparticles into either HTCC or PUL polysaccharides originated clear transparent films. These composite films are transparent in daylight and exhibit visible emission under UV excitation. Photoluminescence studies (emission and excitation spectra and emission decay curves) were performed on the newly prepared films.

Water‐soluble cationic chitosan derivative to improve pigment‐based inkjet printing and antibacterial properties for cellulose substrates

AbstractThe water‐soluble N‐(2‐hydroxy)propyl‐3‐trimethylammonium chitosan chloride (HTCC) was prepared by chitosan and glycidyl trimethyl ammonium chloride. Fourier‐transform infrared spectroscopic analysis spectrum confirmed that glycidyl trimethyl ammonium chloride was grafted to the amino groups of chitosan via substitution reaction (Zhao et al., Int J Pharm 2010, 393, 268; Montazer and Afjeh, J Appl Polym Sci 2007, 103, 178). The obtained chitosan derivative was used to modify cotton fabrics for improving aqueous pigment‐based inkjet printing and antibacterial properties. Scanning electron microscope images showed that HTCC was adhered onto the cotton fabrics surface and formed film structure. The K/S value on printed cotton substrates increased from 3.20 to 4.87, which indicated that higher color yield was achieved in this way. Modified samples performed better crocking fastness and laundering fastness than the control cotton fabrics. The line definition of modified cotton fabrics respectively, improved 16.5% and 12.6% in the warp and weft direction as the specified line width was 0.5 × 103 μm. Samples also showed good antibacterial potential, the inhibitory rate for Staphylococcus aureus and Escherichia coli were both more than 95% when the concentration of HTCC used in the pretreatment solution was 0.8%. The antibacterial effect was found to be durable for 20 laundering cycles. However, the hand feeling of modified cotton fabrics presented a slight decrease. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

Restoration of rat calvarial defects by poly(lactide-co-glycolide)/hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes

A composite construct comprising of a bone mesenchymal stem cell (BMSC) sheet, plasmid DNA, encoding human bone morphogenic protein-2 (hBMP-2), and poly(lactide-co-glycolide)/hydroxyapatite (PLGA/HA) sponge was designed and employed in the restoration of rat calvarial defects. To improve gene transfection efficiency, a cationic chitosan derivative, N,N,N,-trimethyl chitosan chloride (TMC), was employed as the vector. The TMC/DNA complexes had a transfection efficiency of 13% in rat BMSCs, resulting in heterogeneous hBMP-2 expression in a 10-d culture period in vitro. In vivo culture of the composite constructs was performed by implantation into rat full-thickness calvarial defects, using constructs lacking pDNA-hBMP-2 or BMSC sheets as controls. Significantly higher heterogeneous expression of hBMP-2 was detected in vivo at 2 weeks for the cell sheet/DNA complex/scaffold constructs, compared with the constructs lacking pDNA-hBMP-2 or BMSC sheets. New bone formation was evident as early as 4 weeks in the experimental constructs. At 8 weeks, partial bridging of calvarial defects was observed in the experimental constructs, which was significantly better than the constructs lacking pDNA-hBMP-2 or BMSC sheets. Therefore, the combination of the PLGA/HA scaffold with BMSC sheets and gene therapy vectors is effective at enhancing bone formation.

Chitosan Derivatives with Antimicrobial, Antitumour and Antioxidant Activities - a Review

Chitosan is a linear polysaccharide with a good biodegradability, biocompatibility, and no toxicity, which provide it with huge potential for future development. The chitosan molecule appears to be a suitable polymeric complex for many biomedical applications. This review gathers current findings on the antibacterial, antifungal, antitumour and antioxidant activities of chitosan derivatives and concurs with our previous review presenting data collected up to 2008. Antibacterial activity is based on molecular weight, the degree of deacetylation, the type of substitutents, which can be cationic or easily form cations, and the type of bacterium. In general, high molecular weight chitosan cannot pass through cell membranes and forms a film that protects cells against nutrient transport through the microbial cell membrane. Low molecular weight chitosan derivatives are water soluble and can better incorporate the active molecule into the cell. Gram-negative bacteria, often represented by Escherichia coli, have an anionic bacterial surface on which cationic chitosan derivatives interact electrostatically. Thus, many chitosan conjugates have cationic components such as ammonium, pyridinium or piperazinium substituents introduced into their molecules to increase their positive charge. Gram-positive bacteria like Staphylococcus aureus are inhibited by the binding of lower molecular weight chitosan derivatives to DNA or RNA. Chitosan nanoparticles exhibit an increase in loading capacity and efficacy. Antitumour active compounds such as doxorubicin, paclitaxel, docetaxel and norcantharidin are used as drug carriers. It is evident that chitosan, with its low molecular weight, is a useful carrier for molecular drugs requiring targeted delivery. The antioxidant scavenging activity of chitosan has been established by the strong hydrogen-donating ability of chitosan. The low molecular weight and greater degree of quarternization have a positive influence on the antioxidant activity of chitosan. Phenolic and polyphenolic compounds with antioxidant effects are condensed with chitosan to form mutual prodrugs.

Creaming of skim natural rubber latex with chitosan derivatives

AbstractTo recover residual rubber from skim natural rubber (SNR) latex, a novel method was developed on the basis of the use of water‐soluble chitosan derivatives. An anionic chitosan derivative, N,O‐carboxymethyl chitosan (CMCh), and a cationic chitosan derivative, N‐(2‐hydroxy)propyl‐3‐trimethylammonium chitosan chloride (HTACh), were prepared. It was found that the creaming process could be achieved with both chitosan derivatives. By the addition of 7.34–10.3‐g/L CMCh, SNR latex was almost completely creamed, and no coagulation of SNR particles in the cream phase occurred. In the case of HTACh, the creaming process was achieved with a lower concentration (<3.70 g/L) than in the case of CMCh, but the size of SNR particles in the cream phase was larger. Solid‐state 13C‐NMR, thermogravimetric analysis, and ζ potential measurement results provided evidence that the creaming of SNR latex with CMCh and HTACh was controlled by the depletion flocculation and adsorption mechanisms, respectively. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010

Water-Soluble Polyelectrolyte Complexes of Oppositely Charged Polysaccharides

The article is an overview of our recent study on some particular aspects of polyelectrolyte complex (PEC) formation by oppositely charged polysaccharides when they are brought into contact in aqueous solutions. This type of complexation can lead to the thickening effect, jellification or PEC precipitation that find numerous applications in a variety of fields from the regulation of rheological characteristics of solutions to fabrication of functional materials by the layer-by-layer technique. Our focus was on the rheological aspects of water-soluble PEC formation and jellification, but to gain an insight into the mechanisms of the processes involved, atomic force microscopy, scanning electron microscopy and differential scanning calorimetry were also applied. As cationic polysaccharides, chitosan and cationic derivatives of hydroxypropylcellulose including hydrophobically modified samples were taken and, as their anionic counterparts, alginates, carrageenans, xanthans and fucoidans were used. Their combination allowed us to consider the influence of charge density, hydrophobicity and flexibility–stiffness of macromolecules on the association of oppositely charged polysaccharides, the formation of temperature sensitive hydrogels and some PEC morphological features.

Cytotoxic activity of aminoderivatized cationic chitosan derivatives

Chitosan derivatives were prepared by dialkylaminoalkylation and reductive amination followed by quaternization. In this study, the cytotoxic activity of the chitosan derivatives was investigated and a relationship between structure and activity is suggested. The cationic chitosan derivatives elicited dose-dependent inhibitory effects on the proliferation of tumor cell lines.

Modification of chitosan to improve its hypocholesterolemic capacity.

Cholestyramine is the most widely used bile acid sequestrant in the treatment of hypercholesterolemia. However, cholestyramine has unpleasant side effects as a consequence of its hydrophobic backbone. Therefore, high-capacity bile acid sequestering biopolymers with cationic chitosan derivatives were developed, because electrostatic interactions are important for binding with bile acid anions. Dialkylaminoalkylation and reductive amination of chitosan were done to add dialkylaminoalkyl and an additional free amino group at a hydroxyl site in the chitosan backbone respectively and the amino-derivatized chitosan derivatives were quaternized with methyl iodide to produce a cationic polyelectrolyte. The in vitro bile acid binding capacity of the chitosan derivatives in aqueous NaCl was measured by reversed-phase HPLC. The binding capacities of sodium glycocholate (a major bile acid) to chitosan, DEAE-chitosan, quaternized DEAE-chitosan, and cholestyramine were 1.42, 3.12, 4.06, and 2.78 mmol/g resin, respectively. With quaternized DEAE-chitosan, the bile acid binding capacity increased approximately 50% over that of cholestyramine. The bile acid binding capacity of dialkylaminoalkyl chitosan derivatives increased with the number of carbons in the alkyl groups, indicating that hydrophobic interaction is a secondary factor for the sequestration of bile acids.