Quaternized Chitosan Nanoparticles Research Articles

Konjac glucomannan-based nanocomposite spray coating with antimicrobial, gas barrier, UV blocking, and antioxidation for bananas preservation

Fruit is prone to rot and deterioration due to oxidative browning and microbial infection during storage, which can cause serious economic losses and food safety problems. It is urgent to develop a multifunctional composite coating to extend the shelf life of fruits. In this work, multifunctional quaternized chitosan nanoparticles (QCs/TA NPs) with excellent antibacterial and antioxidant properties were prepared based on electrostatic interaction using tannic acid instead of conventional cross-linking agents. Meanwhile, konjac glucomannan (KGM) with high viscosity, edible and biodegradable properties was used as a dispersant to disperse and stabilize the nanoparticles, and as a film-forming agent to form a multifunctional composite coating. The composite coating exhibited excellent oxygen and water vapor barrier properties, antioxidant, antibacterial, mechanical properties, hydrophobicity, and UV shielding properties. Surprisingly, the oxygen permeability of the K-NPs-15 composite film was as low as 1.93 × 10−13 (cm3·cm)/(cm2·s·Pa). The banana spray preservation experiments proved that the K-NPs-15 composite coating could effectively prolong the shelf life of bananas. Therefore, this study provides a new idea for designing multifunctional freshness preservation coatings, which has a broad application prospect.

Exploring the antimicrobial potential of chitosan nanoparticles: synthesis, characterization and impact on Pseudomonas aeruginosa virulence factors.

The escalating antibiotic resistance observed in bacteria poses a significant threat to society, with the global prevalence of resistant strains of Pseudomonas aeruginosa on the rise. Addressing this challenge necessitates exploring strategies that would complement existing antimicrobial agents, e.g. by substances mitigating bacterial virulence without eliciting selective pressure for resistance emergence. In this respect, free-form chitosan has demonstrated promising efficacy, prompting our investigation into reinforcing its effects through nanoparticle formulations. Our study focuses on the preparation of chitosan nanoparticles under suitable conditions while emphasizing the challenges associated with stability that can affect biological activity. These challenges are mitigated by introducing quaternized chitosan, which ensures colloidal stability in the culture media. Our approach led to the production of trimethylchitosan nanoparticles with a median size of 103 nm, circularity of 0.967, and a charge of 14.9 ± 3.1 mV, stable within a one-month period in a water stock solution, showing promising attributes for further valorization. Furthermore, the study delves into the antimicrobial activity of trimethylchitosan nanoparticles on Pseudomonas aeruginosa and confirms the benefits of both nanoformulation and modification of chitosan, as our prepared nanoparticles inhibit 50% of the bacterial population at concentration ≥160 mg L-1 within tested strains. Additionally, we identified a concentration of 5 mg L-1 that no longer impedes bacterial growth, allowing reliable verification of the effect of the prepared nanoparticles on Pseudomonas aeruginosa virulence factors, including motility, protease activity, hemolytic activity, rhamnolipids, pyocyanin, and biofilm production. Although trimethylchitosan nanoparticles exhibit promise as an effective antibiofilm agent (reducing biofilm development by 50% at concentrations ranging from 80 to 160 mg L-1) their impact on virulence manifestation is likely not directly associated with quorum sensing. Instead, it can probably be attributed to non-specific interactions with the bacterial surface. This exploration provides valuable insights into the potential of quaternized chitosan nanoparticles in addressing Pseudomonas aeruginosa infections and underscores the multifaceted nature of their antimicrobial effects.

Preparation and characterization of antibacterial, antioxidant, and biocompatible p-coumaric acid modified quaternized chitosan nanoparticles

To fabricate multifunctional nanoparticles (NPs) based on chitosan (CS) derivative, we first prepared quaternized CS (2-hydroxypropyltrimethyl ammonium chloride CS, HTCC) via a one-step approach, then synthesized p-coumaric acid (p-CA) modified HTCC (HTCC-CA) for the first time through amide reaction, and finally fabricated a series of NPs (HTCC-CA NPs) using HTCC-CAs with different substitution degrees and sodium tripolyphosphate (TPP) by ionic gelation. Newly-prepared HTCC and HTCC-CAs were characterized by FT-IR, 1H NMR, elemental analysis (EA), full-wavelength UV scanning, silver nitrate titration, and Folin-Ciocalteu methods. DLS and TEM results demonstrated that three selected HTCC-CA NPs had moderate size (< 350 nm), good dispersion (PDI < 0.4), and positive zeta potential (11–20 mV). The HTCC-CA NPs had high antibacterial activity against six bacterial strains, and the minimum inhibitory concentration (MIC) values were almost the same as the minimum bactericidal concentration (MBC) values (250–1000 μg/mL). Also, the HTCC-CA NPs had good antioxidation (radical scavenging ratio > 65 %) and low cytotoxicity (relative cell viability >80 %) to the tested cells. Totally, HTCC-CA NPs with high antibacterial activity, great antioxidation, and low cytotoxicity might serve as new biomedical materials for promoting skin wound healing.

Cross-linked quaternized chitosan nanoparticles for effective delivery and controllable release of O. europaea phenolic extract targeting cancer therapy

Cross-linked quaternized chitosan nanoparticles for effective delivery and controllable release of O. europaea phenolic extract targeting cancer therapy

Ketoconazole loaded quaternized chitosan nanoparticles-PVA film: preparation and evaluation

In the present study, ketoconazole an antifungal drug has been entrapped in quaternized chitosan (QCS) nanoparticles. Firstly, the quaternization of chitosan was carried out with the aid of microwave assisted technique, and then, ketoconazole loaded quaternized chitosan nanoparticles (KCZ-QCSNPs) were formed by cross-linking of QCS by sodium tripolyphosphate through ionic gelation method. The quaternization of chitosan was confirmed by proton nuclear magnetic resonance spectroscopy. The KCZ-QCSNPs were characterized with the help of fourier transform infrared spectroscopy, differential scanning calorimetry and X-ray diffraction study. The KCZ-QCSNPs were evaluated for size, shape and distribution with the aid of field-emission scanning electron microscopy. The entrapment efficiency (EE%) of different batches of ketoconazole loaded QCSNPs ranges from 80 to 90%.The anti-microbial activities of the CS, QCS and KCZ-QCSNPs against bacterial and fungal strains were studied by tube dilution method. The performed drug loaded nanoparticles were mixed into a solution of PVA to fabricate medicated PVA films carrying KCZ-QCSNPs and studied the release behavior.In vitro drug release of KCZ-QCSNPs, KCZ-QCSNPs-PVA film and KCZ-PVA film were carried out in PBS (pH 7.4) solution. The in vitro release data fitted into various release kinetic model to study the release behavior.

Quaternized Chitosan Nanoparticles in Vaccine Applications.

Different natural and synthetic biodegradable polymers have been used in vaccine formulations as adjuvant and delivery system but have faced various limitations. Chitosan is a new delivery system with the potential to improve development of nano vaccines and drugs. However, chitosan is only soluble in acidic solutions of low concentration inorganic acids such as dilute acetic acid and dilute hydrochloric acid and in pure organic solvents, which greatly limits its application. Chemical modification of chitosan is an important way to improve its weak solubility. Quaternized chitosan not only retains the excellent properties of chitosan, but also improves its water solubility for a wider application. Recently, quaternized chitosan nanoparticles have been widely used in biomedical field. This review focuses on some quaternized chitosan nanoparticles, and points out the advantages and research direction of quaternized chitosan nanoparticles. As shown by the applications of quaternized chitosan nanoparticles as adjuvant and delivery carrier in vaccines, quaternized chitosan nanoparticles have promising potential in application for the development of nano vaccines in the future.

Curdlan sulfate/O-linked quaternized chitosan nanoparticles acting as potential adjuvants promote multiple arms of immune responses

Nasal immunization to prevent and treat diseases caused by infection through the respiratory tract cannot be actualized because of the lack of effective adjuvants. We have proven that compared with antigens loaded on CS or O-HTCC alone in nasal vaccination, antigens loaded on the nanoparticles of curdlan sulfate/O-(2-hydroxyl) propyl-3-trimethyl ammonium chitosan chloride (CS/O-HTCC) can induce stronger systemic and mucosal immune responses. In this study, we evaluated the immunostimulatory activity and mechanisms of CS/O-HTCC nanoparticles. The results showed that CS/O-HTCC nanoparticles can improve the activation of antigen-presenting cells, upregulate the production of inflammatory factors and cytokines, induce cross-presentation, and simultaneously activate type I interferon-related genes. CS/O-HTCC nanoparticles also activated the PI3K/AKT and MAPK pathways and significantly promoted IL-2 transcription to induce the proliferation of lymphocytes. The results revealed that CS/O-HTCC nanoparticles as a type of multifunctional adjuvant can improve multiple arms of immune responses.

Fabrication of Quaternized Chitosan Nanoparticles Using Tripolyphosphate/Genipin Dual Cross-Linkers as a Protein Delivery System.

Various amounts of 2-((acryloyloxy)ethyl)trimethylammonium chloride were grafted onto chitosan (CS) via redox polymerization method to obtain water-soluble quaternized CS (QCS). The QCS nanoparticles loaded with bovine serum albumin (BSA) were then produced by ionic gelation with tripolyphosphate (TPP) and further covalently cross-linked with genipin. The formation of QCS nanoparticles was optimized as a function of monomer grafting yield, QCS/TPP weight ratio, and QCS/genipin weight ratio by Box-Behnken design and response surface methodology. The results showed that QCS nanoparticles prepared with a grafting yield of 50%, QCS/TPP weight ratio of 7.67, and QCS/genipin weight ratio of 60 had a particle size of 193.68 ± 44.92 nm, polydispersity of 0.232, zeta potential of +23.97 mV and BSA encapsulation efficiency of 46.37 ± 2.89%, which were close to the predicted values from mathematical models. In vitro drug release studies at pH 1.2 and pH 7.4 exhibited that the release rate of BSA was significantly decreased and the release period was significantly prolonged after QCS nanoparticles cross-linking with genipin. Therefore, QCS nanoparticles cross-linked with TPP/genipin dual cross-linkers may be a promising protein drug carrier for a prolonged and sustained delivery.

Curdlan sulfate-O-linked quaternized chitosan nanoparticles: potential adjuvants to improve the immunogenicity of exogenous antigens via intranasal vaccination.

IntroductionThe development of ideal vaccine adjuvants for intranasal vaccination can provide convenience for many vaccinations. As an ideal intranasal vaccine adjuvant, it should have the properties of assisting soluble antigens to pass the mucosal barrier and potentiating both systemic and mucosal immunity via nasal administration.MethodsBy using the advantages of polysaccharides, which can promote both T-helper 1 and 2 responses, curdlan sulfate (CS)–O-(2-hydroxyl)propyl-3-trimethyl ammonium chitosan chloride (O-HTCC) nanoparticles were prepared by interacting CS with O-HTCC, and the adjuvancy of the nanoparticles was investigated.ResultsThe results showed that the polysaccharide-based nanoparticles induced the proliferation and activation of antigen-presenting cells. High protein-loading efficiency was obtained by testing with the model antigen ovalbumin (Ova), and the Ova adsorbed onto the cationic CS/O-HTCC complexes was taken up easily by the epithelium. To evaluate the capacity of the Ova/CS/O-HTCC nanoparticles for immune enhancement in vivo, we collected and analyzed immunocytes, serum, and mucosal lavage fluid from intranasally vaccinated mice. The results showed that Ova/CS/O-HTCC nanoparticles induced activation and maturation of antigen-presenting cells and provoked the proliferation and differentiation of lymphocytes more significantly compared to the immunization of Ova mixed with aluminum hydroxide gel. Furthermore, CS/O-HTCC evoked a significantly higher level of Ova-specific antibodies.ConclusionTherefore, these results suggest that CS/O-HTCC nanoparticles are ideal vaccine adjuvants for soluble antigens used in intranasal or mucosal vaccination.

Self-assembly of quaternized chitosan nanoparticles within nanoclay layers for enhancement of interfacial properties in toughened polymer nanocomposites

Role of montmorillonite nanoclay in self-assembly of quaternized chitosan nanoparticles (QCn) within the clay layers was investigated. Inorganic-organic nanohybrid of clay-chitosan was used as a reactive reinforcing agent in preparation of toughened epoxy nanocomposites. Intercalation of QCn within the clay layers increases d-spacing of clay layers from 1.2nm to 3.6nm, leading to an enhanced exfoliation degree in epoxy nanocomposites. The nanohybrid of clay modified with QCn was then incorporated into an epoxy matrix. It was found that the strong interfacial interactions benefit not only the dispersion of the clay within epoxy but also the effective interfacial stress transfer, leading to significantly improved mechanical properties. Rheological investigations showed that the interfacial interactions between the QCn modified clay and epoxy are dominated by the covalent/hydrogen bondings between the amine/hydroxyl enriched QCn and the epoxy matrix.

Preparation, characterization and biocompatibility of poly(vinyl alcohol) films containing tetracycline hydrochloride-loaded quaternized chitosan nanoparticles

Poly(vinyl alcohol) films are widely used in biomaterials applications due to their excellent chemical and thermal stability, transparency, biocompatibility, nontoxicity, and good film-forming ability. In this study, tetracycline hydrochloride (TC) has been encapsulated in quaternized chitosan nanoparticles (QChNPs) by ionic gelation of QCh with sodium tripolyphosphate (TPP). The success of the encapsulation was confirmed by Fourier transform infrared (FT-IR) spectroscopy. The obtained nanoparticles exhibited a regular distribution and spherical shape, with a size range of 450–800 nm as observed by scanning electron microscopy (SEM). The encapsulation efficiencies (EE) of the TC-loaded QChNPs were approximately 72–95%. The obtained nanoparticles were then added into a PVA solution to fabricate PVA films containing TC-loaded QChNPs. In vitro release studies showed an initial burst effect followed by a slow drug release. The antibacterial activities of the PVA films containing TC-loaded QChNPs against E. coli, S. Aureus, and Ent. faecium were evaluated systematically by the disk diffusion method (AATCC 147) and broth dilution method. Finally, the indirect cytotoxicity of the wound dressings was studied in mouse fibroblast (L929) and human fibroblast cells (FB cells) using an MTT assay.

Quaternized chitosan nanoparticles loaded with the combined attenuated live vaccine against Newcastle disease and infectious bronchitis elicit immune response in chicken after intranasal administration

Newcastle disease (ND) and infectious bronchitis (IB) are important diseases, which cause respiratory diseases in chickens, resulting in severely economic losses in the poultry industry. In this study, N-2-hydroxypropyl trimethyl ammonium chloride chitosan (N-2-HACC) and N,O-carboxymethyl chitosan (CMC) were synthesized as adjuvant and delivery carrier for vaccine antigens. N-2-HACC-CMC/NDV/IBV nanoparticles (NPs) (NDV/La Sota and IBV/H120 encapsulated in N-2-HACC-CMC NPs) and N-2-HACC-CMC/NDV-IBV NPs (the mixing of N-2-HACC-CMC/NDV NPs and N-2-HACC-CMC/IBV NPs in a ratio of 1:1) were prepared by the polyelectrolyte composite method, respectively. Both nanoparticles exhibited lower cytotoxicity and higher stability. Their bioactivities were maintained when they were stored at 37 °C for three weeks. Release assay in vitro showed that both NDV and IBV could be sustainably released from the nanoparticles after an initial burst release. In vivo immunization of chickens showed that N-2-HACC-CMC/NDV/IBV NPs or N-2-HACC-CMC/NDV-IBV NPs intranasally induced higher titers of IgG and IgA antibodies, significantly promoted proliferation of lymphocytes and induced higher levels of interleukine-2 (IL-2), IL-4 and interferon-γ (IFN-γ) than the commercially combined attenuated live vaccine did. This is the first study in the field of animal vaccines demonstrating that intranasal administration of chickens with antigens (NDV and IBV) encapsulated with chitosan derivative could induce humoral, cellular, and mucosal immune responses, which protected chickens from the infection of highly virulent NDV and IBV. This study indicated that N-2-HACC-CMC could be used as an efficient adjuvant and delivery carrier for further development of mucosal vaccines and drugs and could have an immense application potential in medicine.

Cationic Quaternized Chitosan Nanoparticles for C-phycocyanin Delivery to Hela Cells

Cationic Quaternized Chitosan Nanoparticles for C-phycocyanin Delivery to Hela Cells

Novel quaternized polyvinyl alcohol/quaternized chitosan nano-composite as an effective hydroxide-conducting electrolyte

A novel polymer composite electrolyte consisting of quaternized polyvinyl alcohol (Q-PVA) and quaternized chitosan nano-particles (Q-chitosan) is synthesized for use in a direct alcohol alkaline fuel cell (DAAFC). The quaternization efficiencies of PVA and chitosan particles are 2.6% and 39%, respectively. Incorporation of 5% Q-chitosan nano-particles into the Q-PVA matrix results not only in an increased ion-exchange capacity but also a decreased polymer crystallinity and higher free volume hole density, which significantly enhance ion conduction. The shrunk polymer free volume size suppresses methanol permeability in the Q-PVA/Q-chitosan composite. In addition, the resulting nano-composite exhibited an inhibited in-plane swelling ratio without sacrificing the alkali uptake level. The KOH-doped Q-PVA/Q-chitosan was resistant to the Fenton reagent. In DAAFCs, a maximum power density of 73mWcm–2 is achieved using the Q-PVA/5%Q-chitosan electrolyte when fed with 2M methanol at 60°C or 59mWcm–2 with 3M ethanol at 60°C. The resulting maximum power densities are higher than most literature data. The alkaline fuel cell demonstrated stable long-term power output for at least 230h. This result confirms that the Q-PVA/Q-chitosan electrolyte exhibits great potential for future commercialization.

Preparation and testing of quaternized chitosan nanoparticles as gene delivery vehicles.

The aim of this study was to synthesize a chitosan (CS) derivative, a quaternary ammonium salt crystal called N-2-hydroxypropyl trimethyl ammonium chloride chitosan (HACC), and test a series of HACC and pEGFP-DNA complexes at different weight ratios for their efficiency of gene delivery into human cells. CS was modified with cationic etherifying agent to obtain the CS derivative. Fourier transform infrared spectra were recorded on KBr pellets with a spectrometer. (1)H nuclear magnetic resonance (NMR) spectra of HACC were obtained using a spectrometer. HACC was subsequently used to prepare HACC/DNA complexes at different weight ratios by coacervation method. The resulting particle size and surface charge were assessed by laser light scattering using a zeta potential analyzer. The HACC/DNA complex formation and DNA protection in the nanoparticle complex was investigated by gel mobility shift assay and DNase I protection assay, respectively. The cytotoxicity of HACC and HACC/DNA nanoparticles was evaluated by MTT assay using (mesenchymal stem cell) MSC lines. The nanoscale structure of the particles was obtained by transmission electron microscope (TEM). The FTIR spectrum of HACC showed the characteristic quaternary ammonium group absorption band at 1475 cm(-1), which indicated the presence of quaternary ammonium group. The successful synthesis of HACC was also confirmed by (1)H NMR spectrum. HACC showed good solubility in water and was electropositive. HACC efficiently packed and protected pEGFP-DNA at a weight ratio of 10. With increased weight ratios, the surface charge of the composite particle increased from negative to positive, the average particle size increased, and HACC nanoparticle had a higher carrying efficiency. The nanoparticles released DNA in two distinct phases, and 55 % was released within the first 20 h of solubilization. The nanoparticles under TEM showed circular or oval shapes. The particles exhibited no cytotoxicity against human cells. No significant difference in gene delivery efficiency was detected between HACC/pEGFP-GDNF and liposome/pEGFP-GDNF complexes (33.8 vs. 34 %, P = 0.363). In this study, HACC was successfully synthesized, and HACC/DNA complex assembled efficiently. HACC showed strong DNA binding affinity and high protection of DNA and was non-cytotoxic to human cells. The particles had appropriate nanostructure, mean diameter, and DNA release time. The results suggest that HACC nanoparticles are a novel tool for efficient and safe gene delivery.

Alginate-coated quaternized chitosan nanoparticles for oral delivery of insulin

Alginate-coated quaternized chitosan nanoparticles for oral delivery of insulin

Porous Quaternized Chitosan Nanoparticles Containing Paclitaxel Nanocrystals Improved Therapeutic Efficacy in Non-Small-Cell Lung Cancer after Oral Administration

Clinical application of paclitaxel (PTX) is limited because of its poor solubility in aqueous media. To overcome this hurdle, we devised an oral delivery system by encapsulating PTX into N-((2-hydroxy-3-trimethylammonium) propyl) chitosan chloride (HTCC) nanoparticles. These nanoparticles were small (~130 nm), had a narrow size distribution, and displayed high loading efficiency owing to the homogeneous distribution of PTX nanocrystals. The matrix hydrophilicity and porous structure of the obtained nanoparticles accelerated their degradation and improved drug release. In vitro and in vivo transport experiments had proved that the presence of positive charges enhanced the intestinal permeability of these nanoparticles. Further in vitro experiment of cytotoxicity showed that the PTX-loaded HTCC nanoparticle (HTCC-NP:PTX) was more effective than native PTX owing to enhanced cellular uptake. Drug distribution in tissues and in vivo imaging studies confirmed the preferred accumulation of HTCC-NP:PTX in subcutaneous tumor tissue. Subsequent tumor xenograft assays demonstrated the promising therapeutic effect of HTCC-NP:PTX on inhibition of tumor growth and induction of apoptosis in tumor cells. Additional investigation into side effects revealed that HTCC-NP:PTX caused lower Cremophor EL-associated toxicities compared with Taxol. These results strongly supported the notion that HTCC nanoparticle (HTCC-NP) is a promising candidate as an oral carrier of PTX for cancer therapy.

Transmission electron microscopy and electron diffraction study of BSA‐loaded quaternized chitosan nanoparticles

Chitosan nanoparticles, O-(2-hydroxyl) propyl-3-trimethyl ammonium chitosan chloride (O-HTCC) nanoparticles and bovine serum albumin (BSA) loaded chitosan and O-HTCC nanoparticles of a size (about 200-600 nm) were obtained through the process of ionic gelation between chitosan or O-HTCC and sodium tripolyphosphate (TPP). The physicochemical properties of nanoparticles made from chitosan, O-HTCC, BSA loaded chitosan, and BSA loaded O-HTCC were determined by transmission electron microscopy (TEM), polarized optical microscopy (POM), photon correlation spectroscopy (PCS), and X-ray diffraction (XRD) pattern. Zeta potential was also performed to understand the surface properties of nanoparticles and their ability to bind negatively charged BSA. TEM, POM, and XRD suggested that ionic-gelation process significantly influenced the crystallinity of BSA, and greater chain realignment in the BSA-loaded chitosan and O-HTCC nanoparticles. PCS revealed that BSA-loaded chitosan nanoparticles were bigger than chitosan nanoparticles in size and BSA-loaded O-HTCC nanoparticles were smaller than O-HTCC nanoparticles in size.