Blank Chitosan Nanoparticles Research Articles

Synthesis, characterization, and evaluation of the antifungal properties of tissue conditioner incorporated with essential oils-loaded chitosan nanoparticles.

PurposeThis study aims to investigate new tissue conditioner (TC) formulations involving chitosan nanoparticles (CSNPs) and essential oils (EO) for their antifungal potential, release kinetics, and hardness.Materials and methodsCSNPs were synthesized, and the separate solutions of CSNP were prepared with two types of EO, i.e., Oregano oil and Lemongrass. The EO was loaded separately in two concentrations (200 μL and 250 μL). The blank and EO-loaded CSNPs were screened against Candida albicans (C. albicans), and their minimum inhibitory concentration was established. GC Reline™ (GC corporation, USA) TC was considered a control group, whereby the four experimental groups were prepared by mixing CSNPs/EO solutions with TC powder. The antifungal effectiveness (C. albicans) and release kinetics behavior (1–6 h, 24 h, 48 h, and 72 h) was investigated. The Shore A hardness of control and experimental groups was evaluated in dry and wet modes (deionized water and artificial saliva). For statistical analysis, SPSS version 22 was used to do a one-way ANOVA post-hoc Tukey’s test.ResultsCompared to the control group, TCs containing blank CSNPs and CSNPs loaded with EO showed 3 and 5 log reductions in C. albicans growth, respectively. A significantly high antifungal effect was observed with TC containing lemongrass essential oil (200 μL). The continuous release of EO was detected for the first 6 hours, whereas completely stopped after 48 hours. Mean hardness values were highest for dry samples and lowest for samples stored in artificial saliva. The statistically significant difference within and between the study groups was observed in mean and cumulative essential oils release and hardness values of TCs over observed time intervals irrespective of storage media.ConclusionTCs containing essential-oil-loaded CSNPs seem a promising alternative treatment of denture-induced stomatitis, however, a further biological analysis should be taken.

Development and Evaluation of Chitosan Nanoparticles for Ocular Delivery of Tedizolid Phosphate.

This study investigates the development of topically applied non-invasive chitosan-nanoparticles (CSNPs) for ocular delivery of tedizolid phosphate (TZP) for the treatment of MRSA-related ocular and orbital infections. An ionic-gelation method was used to prepare TZP-encapsulated CSNPs using tripolyphosphate-sodium (TPP) as cross-linker. Particle characterization was performed by the DLS technique (Zeta-Sizer), structural morphology was observed by SEM. The drug encapsulation and loading were determined by the indirect method. In-vitro release was conducted through dialysis bags in simulated tear fluid (pH 7) with 0.25% Tween-80. Physicochemical characterizations were performed for ocular suitability of CSNPS. An antimicrobial assay was conducted on different strains of Gram-positive bacteria. Eye-irritation from CSNPs was checked in rabbits. Transcorneal flux and apparent permeability of TZP from CSNPs was estimated through excised rabbit cornea. Ionic interaction between the anionic and cationic functional groups of TPP and CS, respectively, resulted in the formation of CSNPs at varying weight ratios of CS/TPP with magnetic stirring (700 rpm) for 4 h. The CS/TPP weight ratio of 3.11:1 with 10 mg of TZP resulted in optimal-sized CSNPs (129.13 nm) with high encapsulation (82%) and better drug loading (7%). Release profiles indicated 82% of the drug was released from the TZP aqueous suspension (TZP-AqS) within 1 h, while it took 12 h from F2 to release 78% of the drug. Sustained release of TZP from F2 was confirmed by applying different release kinetics models. Linearity in the profile (suggested by Higuchi’s model) indicated the sustained release property CSNPs. F2 has shown significantly increased (p < 0.05) antibacterial activity against some Gram-positive strains including one MRSA strain (SA-6538). F2 exhibited a 2.4-fold increased transcorneal flux and apparent permeation of TZP as compared to TZP-AqS, indicating the better corneal retention. No sign or symptoms of discomfort in the rabbits’ eyes were noted during the irritation test with F2 and blank CSNPs, indicating the non-irritant property of the TZP-CSNPs. Thus, the TZP-loaded CSNPs have strong potential for topical use in the treatment of ocular MRSA infections and related inflammatory conditions.

Control of the Verticillium Wilt on Tomato Plants by Means of Olive Leaf Extracts Loaded on Chitosan Nanoparticles.

In this research, a new ecofriendly and sustainable fungicide agent, with the ability to control Verticillium wilt, was developed. To this purpose, a green extract of olive leaf (OLE) was prepared by ultrasound-assisted extraction (UAE) and characterized in terms of polyphenol content and antioxidant activity. Then, OLE was loaded in chitosan nanoparticles (CTNPs) to combine the antifungal activity of CTNPs and phenolic compounds to obtain an important synergic effect. Nanoparticles were synthetized using the ionic gelation technique and characterized in terms of sizes, polydispersity index, Z-potential, encapsulation efficiency, and release profile. Qualitative and quantitative analyses of OLE were performed by the HPLC method. OLE-loaded CTNPs exhibited good physicochemical properties, such as a small size and positive surface charge that significantly contributed to a high antifungal efficacy against Verticillum dahliae. Therefore, their antifungal activity was evaluated in vitro, using the minimal inhibition concentration (MIC) assay in a concentration range between 0.071 and 1.41 mg/mL. Free OLE, blank CTNPs, and OLE-loaded CTNPs possessed MIC values of 0.35, 0.71, and 0.14 mg/mL, respectively. These results suggest an important synergic effect when OLE was loaded in CTNPs. Thereafter, we tested the two higher concentrations on tomato plants inoculated with V. dahliae, where no fungal growth was observed in the in vitro experiment, 0.71 and 1.41 mg/mL. Interestingly, OLE-loaded CTNPs at the higher concentration used, diminished the symptoms of Verticillium wilt in tomato plants inoculated with V. dahliae and significantly enhanced plant growth. This research offers promising results and opens the possibility to use OLE-loaded CTNPs as safe fungicides in the control strategies of Verticillium wilt at open field.

Fabrication and Characterization of Saffron Stamen Aqueous Extract Controlled Release System as Potential Topical Treatment of Thermal Burn Wounds

AbstractIn our study, we aimed the preparation of Saffron stamen aqueous extract (SSAE) loaded chitosan nanoparticles (CNPs) to be used topically for thermal burns. First, biological activity tests such as DNA binding, DNA cleavage and antibacterial activity were performed to determine the effectiveness of SSAE to be encapsulated. Additionally, a genotoxicity test was performed for the reliability of SSAE. Ionic gelation method was used for encapsulation, and SSAE‐loaded CNPs were obtained. Average particle size, zeta potential, polydispersity index (PdI), morphology, and releasing kinetics of the SSAE‐loaded CNPs and blank CNPs were determined. DNA binding showed that SSAE interacted with DNA electrostatically. It was determined that all doses of SSAE used in hydrolytic and oxidative DNA cleavage experiments cleave DNA. As a result of the Ames / Salmonella test, it was found that 1,25 mg SSAE was not genotoxic. Then, this concentration was used in nanoparticle synthesis. As a result of DLS analysis, it was determined that SSAE‐loaded CNPs have an average size of 57.61±1.234 nm, 0.175±0.008 PdI, and +14±0.755 mV zeta potential. Encapsulation efficiency and loading capacity of SSAE‐loaded CNPs calculated as 71.76 % and 2,64 % respectively. As a result of releasing kinetics, it was determined that 79.54 % of SSAE was released after 168 hours. Antibacterial results revealed that SSAE loaded CNPs were more effective against bacteria than blank SSAE.

Oregano (Origanum vulgar subsp. viride) Essential Oil: Extraction, Preparation, Characterization, and Encapsulation by Chitosan-Carbomer Nanoparticles for Biomedical Application

Background: Plant essential oils (EOs) as natural agents have broad activities, including antibacterial, antifungal, antiviral, insecticidal, and repel activities because of their chemical compositions. Objectives: The objective of this study was to increase the stability of Origanum vulgar subsp. viride EOs by encapsulation in chitosan-carbomer nanoparticles by ionic gelation method. Methods: The EOs from dried leaves of O. vulgar subsp. viride were extracted by hydro-distillation method, and EO components were determined by gas chromatography-mass spectrometry (GC-MS). Besides, OEO-loaded chitosan (CS) nano-capsules were prepared using the ionic gelation method. The molecular structure and morphology of nanoparticles were characterized by Fourier Transform-Infrared (FTIR) and scanning electron microscopy (SEM), respectively. The encapsulation efficiency (EE), loading capacity (LC) of the OEO-loaded CS nanoparticles, and their release profiles were determined using UV/Vis spectrophotometry. Results: The major components of OEO were thymol (20.53%), 4-terpinenol (20.28%), and γ-terpinene (12.22%). The percentages of EE and LC of OEO ranged from 99.25 ± 0.74 to 93.84 ± 0.71 and 38.02 ± 0.18 to 66.73 ± 0.51, respectively, with increasing the OEO to chitosan ratio from 1:0.01 to 1:0.04 W/V. The nanoparticles were regular, uniform, and spherical in shape with an average size of 134 to 181 nm, which were dispersed throughout the solution. The zeta potential values for blank chitosan nanoparticles (CSNPs) and OEO-loaded CSNPs were +23.4 and +38.5 mV, respectively. Conclusions: The results confirmed the suitability of the CS-carbomer complex for OEO- CSNPs formation. It is recommended to evaluate the antimicrobial, insecticidal and insect repel activities of developed OEO nanoparticles in laboratory and field studies.

Polydatin and polydatin-loaded chitosan nanoparticles attenuate diabetic cardiomyopathy in rats.

Hyperglycemia is associated with impairment of heart function. The current study aimed to investigate the ameliorative effect of polydatin-loaded chitosan nanoparticles (PD-CSNPs), polydatin (PD) and metformin (MET) on diabetic cardiomyopathy in rats. Rats divided into six groups; normal-control, diabetic-control, diabetic + CSNPs (diabetic rats treated with 50 mg/kg blank chitosan nanoparticles), diabetic + PD-CSNPs (diabetic rats treated with PD-CSNPs equivalent to 50 mg/kg of polydatin), diabetic + PD (diabetic rats given 50 mg/kg polydatin), diabetic + MET (diabetic rats given 100 mg/kg metformin), orally and daily for 4 weeks. Treatment of diabetic rats with PD-CSNPs, PD and MET showed a significant reduction in the values of glucose and glycosylated hemoglobin with improvement in heart function biomarkers through decreasing serum creatine kinase and creatine kinase myocardial band activities compared to diabetic control. The treatment agents also suppressed the elevated lipid peroxidation product, increased values of glutathione content, superoxide dismutase, superoxide peroxidase, and catalase activities in the heart of diabetic treated rats. Furthermore, PD-CSNPs, PD and MET decreased heart tissue levels of a pro-inflammatory cytokine; tumor necrosis factor-alpha and nuclear factor-kappa β, upregulation of heart gene expressions; nuclear factor erythroid 2-related factor 2 and heme oxygenase-1. Histological and ultrastructural examinations revealed the ameliorative effect of PD-CSNPs, PD and MET against the harmful of diabetic cardiomyopathy by reducing the cardiac fibers, necrotic cardiac myocytes, inflammatory cell infiltration, and the arrangement of the myofibrils and intercalated discs. In conclusion, the new formula of PD-CSNPs was more effective than PD and MET in amelioration the diabetic cardiomyopathy through its antioxidant, anti-inflammatory and prolonged-release properties.

Praziquantel - chitosan nanoformulation against murine Schistosoma mansoni infection

Abstract The aim of the present work was to design praziquantel (PZQ) loaded chitosan nanoparticles (PZQ-Cs) to improve the oral bioavailability and overcome the drawbacks of conventional PZQ therapy. The synthesized chitosan nanoparticles (CsNPs) were physico-chemically characterized by TEM and dynamic light scattering (DLS) with the calculation of the encapsulation efficiency (EE). PZQ was loaded into CsNPs in three different doses; reduced dose (250 mg/kg), high dose (500 mg/kg) and fully effective dose (1000 mg/ kg). Infected treated groups received either oral PZQ-Cs or oral PZQ alone 42 days post-infection and were sacrificed 2 weeks post-treatment. The current study revealed that PZQ-Cs induced a significant anti-schistosomal effect, compared to the infected non-treated control. However, on comparison with conventional PZQ treatment at the corresponding doses, oral PZQ-Cs showed a lower anti-schistosomal potential as reflected by the total worm burden, tissue ova count and oogram pattern. PZQ-Cs was significantly effective in attenuating the oxidative insult associated with S. mansoni infection, compared to the infected non-treated and PZQ treated groups. Genotoxicity assessment proved the safety of CsNPs, as they did not induce a significant DNA fragmentation in non-infected mice treated with blank CsNPs. The present work highlights CsNPs as safe, effective anti-inflammatory and antioxidant agents that could find a clinical use in the treatment of schistosomiasis induced oxidative stress and hepatic dysfunction. Further studies are recommended, however, to investigate how PZQ-Cs could be further modified to increase its anti-schistosomal potential.

Fabrication of chitosan nanoparticles loaded with aromatase inhibitors for the advancement of gonadal development in Clarias magur (Hamilton, 1822)

Aromatase inhibitors (AIs) block the conversion of testosterone to 17β-estradiol and thereby increase the serum testosterone levels. The present work was aimed for development and characterization of nanoconjugated AIs for studying the effect on the gonadal development in Clarias magur. Two AIs such as Fadrozole and Letrozole were used in this study and conjugated with chitosan nanoparticles. The size and zeta potential of the blank chitosan nanoparticles was 47.6 nm and 29 mV, respectively. The size of chitosan conjugated Letrozole (LZC) was 108.9 nm with a zeta potential of 18.9 mV while chitosan conjugated Fadrozole (FZC) showed the size of 150.5 nm with a zeta potential of 25.6 mV. Entrapment efficiency of LZC and FZC were 86.6% and 84.3%, respectively. The FTIR analysis confirmed the conjugation between chitosan and AIs. Fish receiving an intramuscular injection of LZC showed a higher level of serum testosterone than FZC injected fishes. On the other hand, a lower level of 17β-estradiol was observed in LZC treated fish. These results indicate the higher efficiency of letrozole over fadrozole. The oral delivery of Letrozole alone and conjugated with chitosan nanoparticles was carried out through feed (100 mg/kg of feed) to fingerlings. The LZC treated group showed advancement in the testicular development which was evident in serum hormonal analysis, histological examination of gonads and Gonadosomatic Index (GSI). Thus, the nano-formulation developed in the present study may be useful for improving the reproductive output in fish.

Antibacterial Activity of Fructus forsythia Essential Oil and the Application of EO-Loaded Nanoparticles to Food-Borne Pathogens.

Fructus forsythia essential oil (FEO) with excellent antibacterial activity was rarely reported. The objective of the present study was to investigate the antibacterial activity and the antibacterial mechanism of FEO against two food-borne pathogenic bacteria, Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) in vitro. When treated FEO, the zones of inhibition (ZOI) of E. coli (20.5 ± 0.25 mm) and S. aureus (24.3 ± 0.21 mm) were much larger than control (p < 0.05). The minimum inhibitory concentrations (MICs) of FEO were 3.13 mg/mL and 1.56 mg/mL for E. coli and S. aureus, respectively. The antibacterial mechanism of FEO against E. coil was due to the changes in permeability and integrity of cell membrane leading to the leakage of nucleic acids and proteins. With the superior antibacterial activity of FEO, the nano-encapsulation method has been applied in FEO. When compared to FEO and blank chitosan nanoparticles, FEO-loaded nanoparticles (chitosan to FEO of 1:1) can effectively inhibit the growth of E. coil above 90% at room temperature. It is necessary to consider that FEO and FEO-loaded nanoparticles will become promising antibacterial additives for food preservative, cosmetic, and pharmaceutical applications.

Surface-Modified Nanoparticles Enhance Transurothelial Penetration and Delivery of Survivin siRNA in Treating Bladder Cancer

Penetration of the bladder permeability barrier (BPB) is a major challenge when treating bladder diseases via intravesical delivery. To increase transurothelial migration and tissue and tumor cell uptake, poly(lactic-co-glycolic acid; PLGA) nanoparticles (NP) were modified by addition of a low molecular weight (2.5 or 20 kDa) positively charged mucoadhesive polysaccharide, chitosan, to the NP surface. In designing these NPs, we balanced the adhesive properties of chitosan with the release and bioactivity of the siRNA. Chitosan-functionalized NPs demonstrated increased binding to and uptake in intravesically instilled mouse bladders and human ureter at 10 times the level of unmodified NPs. Furthermore, we extended the bioactivity of survivin siRNA in vitro for up to 9 days and demonstrated a decrease in proliferation when using chitosan-modified NPs relative to unmodified NPs. In addition, treatment of xenograft tumors with chitosan-modified NPs that encapsulate survivin siRNA (NP-siSUR-CH2.5) resulted in a 65% reduction in tumor volume and a 75% decrease in survivin expression relative to tumors treated with blank chitosan NPs (NP-Bk-CH2.5). Our low molecular weight chitosan delivery system has the capacity to transport large amounts of siRNA across the urothelium and/or to the tumor site, thus increasing therapeutic response.

The chitosan-small interfering RNA suppressed the expression levels of multidrug resistant-associated protein 1 in C6 cells

Objective To investigate the chitosan-small interfering RNA (siRNA) NP ratio with maximum transfection efficiency and silence effect.Methods The diameter and Zeta potential of the chitosan-siRNA nanoparticles were measured by using laser particle size analyzer.Fluorescent-labeled siRNA was transfected into C6 cells for preliminary appraisal transfection efficiency.The transfected C6 cells were divided into 6 groups:three experimental groups,which were transfected with chitosan-siRNA nanoparticle ratio of 175∶ 1,88∶1 and 44∶ 1,the positive control group,which was transfected with multidrug resistantassociated protein 1 (MRP1)-siRNA by LipofectamineTM 2000 and MRP1 siRNA; the negative control group,which was transfected with chitosan-negative control siRNA nanoparticles,and the blank group,which was transfected with blank chitosan-nanoparticles.The expression levels of MRP1 mRNA and protein in C6 cells were assessed by using reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting respectively.Results The chitosan-siRNA nanoparticles were successfully prepared.With decreases in NP ratio,the chitosan-siRNA nanoparticles had the diameters of 186.7 nm,209.5 nm,and 492.3 am,and the potentials were 18.57 mV,13.52 mV,and 48.58 mV,respectively.Target protein expression could be down-regulated to (23.2 ± 4.3)％ when the NP ratio of nanoparticles was 175.Conclusion In C6 cells,MRP1 can be effectively suppressed when the chitosan-siRNA ratio was 175∶ 1. Key words: Chitosan; Small interfering RNA; Multidrug resistant-associated protein 1

Design and characterization of antitumor drug paclitaxel-loaded chitosan nanoparticles by W/O emulsions

Chitosan nanoparticles and paclitaxel loaded chitosan nanoparticles were prepared by emulsification-crosslinking method in a W/O emulsion system, using glutaraldehyde as crosslinking agent. The mean diameter of chitosan nanoparticles decreased with increase of pH value of the reaction system from 4.5 to 6.5, and increased when the pH exceeded 6.5. Ultraviolet spectrum analysis showed that the largest loading efficiency and encapsulation efficiency could be 8.55% and 94.01%, respectively. In vitro drug release profile was also determined by ultraviolet spectrometry. MTT assays revealed that the blank chitosan nanoparticles had almost none toxicity, and cell culture was carried out accordingly.

Development and validation of an improved Bradford method for determination of insulin from chitosan nanoparticulate systems

Context: Blank chitosan nanoparticles are currently used as reference for the calibration curve, which fails to resolve the supernatant of the nanoparticles in the interference of Coomassie Brilliant Blue G-250 reagent; supernatants are generated at different chitosan nanoparticulate prescriptions, which have different interferences. There are notable errors in the experimental results, and the method is not feasible.Objective: In this study, an improved, rapid, and economic Bradford method was developed and validated.Materials and methods: The pH of the supernatant of blank chitosan nanoparticles was adjusted to 7–9 through adding saturated NaOH. The precipitation (free chitosan) in the solution was separated by centrifuging for about 10 min (4000 r/min).Results: The method eliminated the interference of free chitosan of different prescriptions. The results showed that the method presented a linearity in the range of 50–300 μg/mL (R2 = 0.9992), and possessed a good inter-day and intra-day precision based on relative standard deviation values (less than 3.10%). Recovery of the supernatant of blank chitosan nanoparticles was between 98.30 and 99.93%, and the recovery of blank chitosan nanoparticles was between 95.57 and 100.27%.Discussion and conclusion: The method was further tested for determination of the association efficiency of insulin to nanoparticulate carriers composed of chitosan. Encapsulant release under simulated gastrointestinal fluids was evaluated.

Immunological Effects and Membrane Interactions of Chitosan Nanoparticles

The objective of this study was to investigate the in vitro and in vivo effects of blank chitosan nanoparticles on various molecular markers such as nitric oxide (NO) production, IL-6 gene expression, and lymphocyte proliferation involved in the wound healing process. In addition, the membrane effects of chitosan nanoparticles were evaluated using phospholipid vesicles as a model membrane. Peripheral blood mononuclear cells (PBMC) were treated with blank chitosan nanoparticles, and the effect on NO production, IL-6 gene expression, and lymphocyte proliferation was evaluated. It was observed that IL-6 gene expression was not induced at any of the doses used; however, a statistically significant dose-dependent increase in NO production was observed at doses above 68.18 microg/mL equivalent to chitosan. Furthermore, chitosan nanoparticles showed a statistically significant and dose-dependent lymphocyte proliferation as compared to the control (P < 0.05). It was observed that blank chitosan nanoparticles resulted in strong membrane perturbation when evaluated by differential scanning calorimetry studies. The in vivo effects of the blank chitosan nanoparticles were evaluated using a wound healing model. Blank chitosan nanoparticles showed significantly higher NO production in vivo as compared to the control. Overall, the study clearly indicates the immunoactivating nature of chitosan nanoparticles and their strong membrane interactive potential.