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In recent years, the use of cationic peptides as alternative drugs with anticancer activity has received attention. In this study, the targeted release of curcumin (Cur) and CM11 peptide alone and together against hepatocellular carcinoma (HCC) was evaluated using chitosan nanoparticles (CS NPs) coated with Pres1 that target the SB3 antigen of HCC cells (PreS1-Cur-CM11-CS NPs). SB3 protein is the specific antigen of HCC and the PreS1 peptide is a part of the hepatitis B antigen, which can specifically bind to the SB3 protein. Chitosan was used to prepare NPs. To Cur and CM11 loading, drugs were added to the CS solution in appropriate concentrations. Pres1 was coupled to the surface of the NPs using EDC catalyst to target NPs against HepG2 cells. SEM and DLS analysis confirmed that the PreS1-Cur-CM11-CS NPs had a size of about 132 nm, the ideal size for penetrating the cell membrane. The loading of Cur and CM11 was equal to 87% and 65%, respectively, which had a sustained and better release in the acidic environment than in the physiological environment. The MTT assay showed that PreS1-Cur-CM11-CS NPs act in a targeted and specific manner with the highest toxicity on the HepG2 cells compared to the control by a decrease in viability of about 26% after 48 h based on cell apoptosis. The results showed that PreS1-Cur-CM11-CS NPs are capable of targeted and specific drug release against HepG2 cancer cells and have significant potential to fight this cancer. [ABSTRACT FROM AUTHOR]

Summary: The objective of this study was to develop Tricholoma fracticum extract‐loaded chitosan nanoparticles (TFNPs) by ionic gelation method and to evaluate their in vitro antioxidant activity. Phenolic and flavonoid contents in the T. fracticum extract were measured spectrophotometrically and chromatographically. Characterisation of NPs was evaluated by field emission scanning electron microscopy (FE‐SEM), transmission electron microscopy (TEM), ZETA analysis, Fourier‐transform infrared spectroscopy (FT‐IR), UV–visible spectroscopy (UV–Vis) and thermogravimetric analysis (TGA). In vitro antioxidant capacity was determined using 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) and 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) assays. The phenolic and flavonoid contents in the T. fracticum extract were measured as 7.1 ± 0.3 mg Gallic Acid Equivalent/g extract and 5.5 ± 0.6 mg Quercetin Equivalent/g extract, respectively. The particle size, polydispersity index (PDI) and zeta potential of Blank NP1 and TFNP were 265.5 ± 15.8 nm, 0.4, 38.7 ± 4.0 mV and 333.2 ± 16.3 nm, 0.4, 37.0 ± 4.1 mV, respectively. The highest antioxidant activity was observed in TFNP, followed by T. fracticum extract, chitosan and blank NP, respectively. The preserved or enhanced antioxidant activity observed in the encapsulated T. fracticum extract indicates the potential for loading similar mushroom extracts onto chitosan and thus preserving their bioactive properties. [ABSTRACT FROM AUTHOR]

Aim and Objectives: The aim of the study was to compare and evaluate the push-out bond strength (POBS) of epoxy resin-based and calcium silicate-based sealers with and without incorporating chitosan nanoparticles (CSNPs). Methodology: Eighty human mandibular premolars with a single canal were taken. A standard root length of 15 mm was established. Canals were instrumented with Neoendo files up to size 30, 4% and were irrigated with 5 ml of 2.5% NaOCl, and 5 ml of ethylenediaminetetraacetic acid 17%. Then, samples were obturated using the size 30, 4% gutta-percha, single-cone technique using respective sealers. For POBS, it was grouped as Group 1 Adseal, Group 2 CeraSeal, Group 3 Adseal + 2% wt/vol CSNP, and Group 4 CeraSeal + 2% wt/vol CSNP. Samples embedded in acrylic resin were sectioned transversely at 11 and 7 mm from the apex. Dislodgment resistance was calculated using a universal testing machine. The data were analyzed using a one-way ANOVA test, followed by Tukey's post hoc analysis. The level of significance (P value) was set at P < 0.05. Results: At 7 mm from the apex, Group 4 (CeraSeal + 2% wt/vol CSNP) demonstrated significantly higher mean POBS. However, no significant difference was observed between Groups 1 (Adseal), Group 2 (CeraSeal), and Group 3 (Adseal + 2% wt/vol CSNP). At 11 mm from the apex, Group 2 (CeraSeal) and Group 4 (CeraSeal + 2% wt/vol CSNP) demonstrated significantly higher mean POBS. On comparison between POBS at 7 mm and 11 mm from the apex, the mean POBS at the 11 mm region from the apex was significantly higher in all study groups, which was statistically significant. Conclusion: Within the limitations of the study, it has been concluded that Group 4 (CeraSeal + 2% wt/vol CSNP) has better bond strength compared to the other groups. [ABSTRACT FROM AUTHOR]

Diabetic ketoacidosis (DKA) is regarded to be a communal complication of both type 1 and type 2 diabetes mellitus in children and adolescents. Successful therapy of DKA in children requires prompt diagnosis, strict monitoring of medical indicators, and prompt action. Thymoquinone (Tq) from black cumin loaded chitosan nanoparticles (ChNPs) intend to assess an effective agent to overcome this problem. XRD, FTIR, SEM, and TEM were used in the physicochemical analysis. Enzymatic activity of α-amylase and α-glucosidase was used in in vitro tests of anti-diabetic efficacy. Protecting insulin against enzyme breakdown is a crucial part of the insulin delivery mechanism. In the STZ-induced diabetes RIN-5F cell line, the anti-apoptotic capability of Tq-ChNPs was demonstrated through the NF-κB mediated apoptotic pathway. The combination of thymoquinone and chitosan NPs demonstrated that a wide variety of incredibly effective substances to elevate their curative effects, thus contributing to the growth of clinical and pharmaceutical fields.

The COVID-19 outbreak was a global pandemic with wide-ranging healthcare implications. Although several mRNA-based vaccines delivered using lipid nanoparticles (LNP) have been approved and demonstrated efficacy at reducing the severity and spread of infection, continued rapid viral evolution and disadvantages currently associated with LNP delivery vehicles (such as toxicity) are driving the design of next-generation SARS-CoV-2 vaccines. Herein, we describe the development of a trimethylated chitosan-based nanoparticle layer-by-layer (LbL) delivery platform for multiple antigens as a scalable and safe COVID-19 vaccine, known as, "LbL-CoV19". These vaccine candidates have been demonstrated to be biocompatible, safe, and effective at stimulating both humoral and cellular responses for protection in preclinical studies. Preliminary results also indicate that LbL-CoV19 can potentially achieve rapid, long-lasting, and broad protection against the SARS-CoV-2 challenge. The "plug-and-play" platform technology is well suited to preparedness for future pandemics and disease outbreaks. [ABSTRACT FROM AUTHOR]

Objective To construct the chitosan/BMP-2 plasmid thermosensitive hydrogel complex CS/CSn（pDNA-BMP2）-GP，and evaluate the effect of this composite system on alveolar bone regeneration in beagle dogs. Methods Periodontitis models were established in the second and third premolars from 4 beagle dogs. All teeth were randomly divided into three groups： CS/CSn-GP group，CS/CSn（pDNA-BMP2）-GP group and blank control group. In the CS/CSn-GP group，CS/CSn-GP was injected. In the CS/CSn（pDNA-BMP2）-GP group，CS/CSn（pDNA-BMP2）-GP was injected. In the blank control group，no material was injected. All animals were sacrificed 8 weeks later. The alveolar bone regeneration in periodontitis was observed by Masson staining. The expression of alkaline phosphatase （ALP） in bone defects was observed by calcium and cobalt staining. Results Masson staining showed that different degrees of alveolar bone regeneration was noted in the CS/CSN-GP and CS/CSn（pDNA-BMP2）-GP groups，and significant regeneration was noted in the CS/CSn（pDNA-BMP2）-GP group. Calcium and cobalt staining revealed strongly-positive ALP in the CS/CSn（pDNA-BMP2）-GP group，weakly-positive ALP in the CS/ CSN-GP group，and negative result in the blank control group. The average optical density value in the ALP-positive staining area in the CS/CSn（pDNA-BMP2）-GP group was higher than that in the CS/CSn-GP group，followed by that in the blank control group（all P < 0.05）. Conclusions CS/CSn-GP composite repair system exerts positive effect on alveolar bone regeneration. Inclusion of pDNA-BMP2 can significantly promote the effect on alveolar bone regeneration.

Synergistic immunotherapy of immune checkpoint blockade (ICB) and immunogenic cell death (ICD) has shown remarkable therapeutic efficacy in various cancers. However, patients show low response rates and undesirable outcomes to these combination therapies owing to the recycling mechanism of programmed death-ligand 1 (PD-L1) and the systemic toxicity of ICD-inducing chemotherapeutic drugs. Herein, we propose all-in-one glycol chitosan nanoparticles (CNPs) that can deliver anti-PD-L1 peptide (PP) and doxorubicin (DOX) to targeted tumor tissues for a safe and more effective synergistic immunotherapy. The PP-CNPs, which are prepared by conjugating ᴅ-form PP (NYSKPTDRQYHF) to CNPs, form stable nanoparticles that promote multivalent binding with PD-L1 proteins on the targeted tumor cell surface, resulting in effective lysosomal PD-L1 degradation in contrast with anti-PD-L1 antibody, which induces recycling of endocytosed PD-L1. Consequently, PP-CNPs prevent subcellular PD-L1 recycling and eventually destruct immune escape mechanism in CT26 colon tumor-bearing mice. Moreover, the ICD inducer, DOX is loaded into PP-CNPs (DOX-PP-CNPs) for synergistic ICD and ICB therapy, inducing a large number of damage-associated molecular patterns (DAMPs) in targeted tumor tissues with minimal toxicity in normal tissues. When the DOX-PP-CNPs are intravenously injected into CT26 colon tumor-bearing mice, PP and DOX are efficiently delivered to the tumor tissues via nanoparticle-derived passive and active targeting, which eventually induce both lysosomal PD-L1 degradation and substantial ICD, resulting in a high rate of complete tumor regression (CR: 60%) by a strong antitumor immune response. Collectively, this study demonstrates the superior efficacy of synergistic immunotherapy using all-in-one nanoparticles to deliver PP and DOX to targeted tumor tissues.

ABSTRACT: Non-typhoidal Salmonella infection is a significant health and economic burden in poultry industry. Developing an oral vaccine to induce robust mucosal immunity in the intestines of birds, especially cross protection against different Salmonella serotypes is challenging. Therefore, a potent oral vaccine platform that can mitigate different serotypes of Salmonella is warranted for the poultry industry. We reported earlier that the Salmonella enteritidis (SE) immunogenic outer membrane proteins (OMPs) and flagellin (FLA) entrapped in mannose chitosan nanoparticles (OMPs-FLA-mCS NPs) administered prime-boost (d-3 and 3-wk later) by oral inoculation elicits mucosal immunity and reduces challenge SE colonization by over 1 log10 CFU in birds. In this study, we sought to evaluate whether the SE antigens containing OMPs-FLA-mCS NPs vaccine induces cross-protection against Salmonella typhimurium (ST) in broilers. Our data indicated that the OMPs-FLA-mCS NPs vaccine induced higher cross-protective antibody responses compared to commercial Poulvac ST vaccine (contains a modified-live ST bacterium). Particularly, OMPs-FLA-mCS-NP vaccine elicited OMPs and FLA antigens specific increased production of secretory IgA and IgY antibodies in samples collected at both post-vaccination and post-challenge timepoints compared to commercial vaccine group. Notably, the vaccine reduced the challenge ST bacterial load by 0.8 log10 CFU in the cecal content, which was comparable to the outcome of Poulvac ST vaccination. In conclusion, our data suggested that orally administered OMPs-FLA-mCS-NP SE vaccine elicited cross protective mucosal immune responses against ST colonization in broilers. Thus, this candidate vaccine could be a viable option replacing the existing both live and killed Salmonella vaccines for birds.

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Background: This study evaluated the effect of using chitosan, nano-chitosan, and ethylenediaminetetraacetic acid (EDTA) as final irrigating solutions on smear layer cleanliness and Ca/P ratio of dentin. Methodology: Forty-eight decoronated human single-rooted teeth were used. They were divided randomly into four groups (n = 12) based on the final irrigating solution used as follows: (a) control group (IA; n = 6) normal saline, (IB; n = 6) were left unprepared; group II - 0.2% chitosan; group III - 0.2% nano-chitosan; and group IV - 17% EDTA. Samples were prepared using ProTaper Next and irrigated with 2.6% NaOCl 5 ml after each instrument using 31-gauge needle. Final rinse was used 5 ml/3 min according to the assigned group. The specimens were prepared for evaluation. Results: Best smear layer removal was observed in group IV. No statistically significant differences (P > 0.05) were observed between the experimental groups (II, III, and IV) coronally; however, a statistically significant difference (P < 0.05) was observed between groups II and IV at middle and apical thirds. Intragroup comparison showed that apical third exhibited the highest mean smear layer score among all experimental groups. The highest mean Ca/P ratio was in the 0.2% nano-chitosan group, while the highest calcium loss was in the 17% EDTA group. Conclusions: 17% EDTA is a potent chelating agent that can successfully remove the smear layer but compromises the Ca/p ratio of dentin. However, 0.2% chitosan and its nanoparticles have comparable chelating effects and induce remineralization of the root canal dentin.

The COVID-19 outbreak was a global pandemic with wide-ranging healthcare implications. Although several mRNA-based vaccines delivered using lipid nanoparticles (LNP) have been approved and demonstrated efficacy at reducing the severity and spread of infection, continued rapid viral evolution and disadvantages currently associated with LNP delivery vehicles (such as toxicity) are driving the design of next-generation SARS-CoV-2 vaccines. Herein, we describe the development of a trimethylated chitosan-based nanoparticle layer-by-layer (LbL) delivery platform for multiple antigens as a scalable and safe COVID-19 vaccine, known as, “LbL-CoV19”. These vaccine candidates have been demonstrated to be biocompatible, safe, and effective at stimulating both humoral and cellular responses for protection in preclinical studies. Preliminary results also indicate that LbL-CoV19 can potentially achieve rapid, long-lasting, and broad protection against the SARS-CoV-2 challenge. The “plug-and-play” platform technology is well suited to preparedness for future pandemics and disease outbreaks.

The influence of ultrasonic processing parameters including reaction temperature (60, 70 and 80 °C), time (0, 15, 30, 45 and 60 min) and amplitude (70, 85 and 100%) on the formation and antioxidant activity of Maillard reaction products (MRPs) in a solution of chitosan and glucose (1.5 wt% at mass ratio of 1:1) was investigated. Selected chitosan-glucose MRPs were further studied to determine the effects of solution pH on the fabrication of antioxidative nanoparticles by ionic crosslinking with sodium tripolyphosphate. Results from FT-IR analysis, zeta-potential determination and color measurement indicated that chitosan-glucose MRPs with improved antioxidant activity were successfully produced using an ultrasound-assisted process. The highest antioxidant activity of MRPs was observed at the reaction temperature, time and amplitude of 80 °C, 60 min and 70%, respectively, with ∼ 34.5 and ∼20.2 μg Trolox mL−1 for DPPH scavenging activity and reducing power, respectively. The pH of both MRPs and tripolyphosphate solutions significantly influenced the fabrication and characteristics of the nanoparticles. Using chitosan-glucose MRPs and tripolyphosphate solution at pH 4.0 generated nanoparticles with enhanced antioxidant activity (∼1.6 and ∼ 1.2 μg Trolox mg−1 for reducing power and DPPH scavenging activity, respectively) with the highest percentage yield (∼59%), intermediate particle size (∼447 nm) and zeta-potential ∼ 19.6 mV. These results present innovative findings for the fabrication of chitosan-based nanoparticles with enhanced antioxidant activity by pre-conjugation with glucose via the Maillard reaction aided by ultrasonic processing.

Chitosan is a fibrous compound derived from chitin, which is the second most abundant natural polysaccharide and is produced by crustaceans, including crabs, shrimps, and lobsters. Chitosan has all of the important medicinal properties, including biocompatibility, biodegradability, and hydrophilicity, and it is relatively nontoxic and cationic in nature. Chitosan nanoparticles are particularly useful due to their small size, providing a large surface-to-volume ratio, and physicochemical properties that may differ from that of their bulk counterparts; thus, chitosan nanoparticles (CNPs) are widely used in biomedical applications and, particularly, as contrast agents for medical imaging and as vehicles for drug and gene delivery into tumors. Because CNPs are formed from a natural biopolymer, they can readily be functionalized with drugs, RNA, DNA, and other molecules to target a desired result in vivo. Furthermore, chitosan is approved by the United States Food and Drug Administration as being Generally Recognized as Safe (GRAS). This paper reviews the structural characteristics and various synthesis methods used to produce chitosan nanoparticles and nanostructures, such as ionic gelation, microemulsion, polyelectrolyte complexing, emulsification solvent diffusion, and the reverse micellar method. Various characterization techniques and analyses are also discussed. In addition, we review drug delivery applications of chitosan nanoparticles, including for ocular, oral, pulmonary, nasal, and vaginal methodologies, and applications in cancer therapy and tissue engineering. [ABSTRACT FROM AUTHOR]

Background: This study evaluated the effect of using chitosan, nano-chitosan, and ethylenediaminetetraacetic acid (EDTA) as final irrigating solutions on smear layer cleanliness and Ca/P ratio of dentin. Methodology: Forty-eight decoronated human single-rooted teeth were used. They were divided randomly into four groups (n = 12) based on the final irrigating solution used as follows: (a) control group (IA; n = 6) normal saline, (IB; n = 6) were left unprepared; group II - 0.2% chitosan; group III - 0.2% nano-chitosan; and group IV - 17% EDTA. Samples were prepared using ProTaper Next and irrigated with 2.6% NaOCl 5 ml after each instrument using 31-gauge needle. Final rinse was used 5 ml/3 min according to the assigned group. The specimens were prepared for evaluation. Results: Best smear layer removal was observed in group IV. No statistically significant differences (P > 0.05) were observed between the experimental groups (II, III, and IV) coronally; however, a statistically significant difference (P < 0.05) was observed between groups II and IV at middle and apical thirds. Intragroup comparison showed that apical third exhibited the highest mean smear layer score among all experimental groups. The highest mean Ca/P ratio was in the 0.2% nano-chitosan group, while the highest calcium loss was in the 17% EDTA group. Conclusions: 17% EDTA is a potent chelating agent that can successfully remove the smear layer but compromises the Ca/p ratio of dentin. However, 0.2% chitosan and its nanoparticles have comparable chelating effects and induce remineralization of the root canal dentin. [ABSTRACT FROM AUTHOR]

Starch-based nanocomposite films containing chitosan nanoparticles (S/CNP films) are biodegradable and promising alternatives for non-biodegradable synthetic plastics. Nonetheless, limited work has been conducted to investigate the biodegradability of the films in soil. Thus, this work is aimed at investigating the biodegradation of starch-based films containing different concentrations of CNP (0, 10, 15, 20, 25% w/w solid starch) via a soil burial test using compost and planting soils. The biodegradability was investigated in terms of weight loss, visual appearance, morphology, and structural changes. It was found that S/CNP films biodegraded slower than neat starch films, and the degradation rate was reduced by 46 and 44% in compost and planting soils, respectively, with the increase in the concentrations of CNP from 10 to 25% w/w incorporated into the films. The degradation rate of films in compost soil after 8 days was found to be higher (0.0617 g/day) than in planting soil (0.0266 g/day). Visual appearance, morphology, and structural change results also supported these findings. The biodegradable S/CNP films can be used to reduce the usage of synthetic plastic. [ABSTRACT FROM AUTHOR]

Introduction: MicroRNAs (miRNAs) are short-sequence RNAs that regulate gene expression by targeting messenger RNAs (mRNAs). Recent studies reveal that miRNA-324-5p plays an important role in worsening the ovarian cancer prognosis when the expression is very high. This study aimed to develop a miRNA targeted therapy by targeting the miRNA-324-5p function as a miRNA-324-5p inhibitor. Methods: Chitosan nanoparticles were used for antimiRNA-324-5p delivery into SKOV3 cell lines formulated by ionic gelation method. Antiproliferative effect of CS-NPs-antimiRNA was assessed by the MTT Assay. A mechanism study assessed the anticancer effect of the formula. In silico analysis used miRTar.Human and StarmiRDB combined with Genecard to predict the target genes of antimiR. Hawkdock web server was used to analyze protein-protein interactions that were further validated by quantitative polymerase chain reaction (qPCR). Results: The results of qPCR analysis showed endogenous miRNA-324-5p decreased after 24-hour transfection of antagonist miRNA. Furthermore, the MTT assay results showed that antimiRNA was able to inhibit SKOV3 cell proliferation (80 nM 68.13%, P

The rising prevalence of impaired wound healing and the consequential healthcare burdens have gained increased attention over recent years. This has prompted research into the development of novel wound dressings with augmented wound healing functions. Nanoparticle (NP)-based delivery systems have become attractive candidates in constructing such wound dressings due to their various favourable attributes. The non-toxicity, biocompatibility and bioactivity of chitosan (CS)-based NPs make them ideal candidates for wound applications. This review focusses on the application of CS-based NP systems for use in wound treatment. An overview of the wound healing process was presented, followed by discussion on the properties and suitability of CS and its NPs in wound healing. The wound healing mechanisms exerted by CS-based NPs were then critically analysed and discussed in sections, namely haemostasis, infection prevention, inflammatory response, oxidative stress, angiogenesis, collagen deposition, and wound closure time. The results of the studies were thoroughly reviewed, and contradicting findings were identified and discussed. Based on the literature, the gap in research and future prospects in this research area were identified and highlighted. Current evidence shows that CS-based NPs possess superior wound healing effects either used on their own, or as drug delivery vehicles to encapsulate wound healing agents. It is concluded that great opportunities and potentials exist surrounding the use of CSNPs in wound healing.

Based on the recent research, the natural polysaccharide rich biopolymer of chitosan is an important carrier molecule for drug loading. In addition, the plant essential oils were another important material that frequently used to treat various infectious diseases. In addition, the chitosan loaded essential oils were heightened recent years due to the excellent biomedical properties. In this regard, the current study result was concentrated with encapsulation of plant essential oils into the chitosan nanoparticles for inhibit against A549 human lung cancer cells. In result, the unique and uniform size morphology of chitosan was clearly shown in scanning electron microscope (SEM) and transmission electron microscope (TEM). Further, the IC50 dose of the chitosan loaded essential oils at 1000 µg/mL concentration against A549 lung cancer cells was confirmed using cytotoxicity assay and this concentration was very effective than chitosan and essential oils alone. At 1000 µg/mL concentration, the high proliferation efficiency with unclear morphological evidence of A549 lung cancer cells was shown in phase contrast microscope. In addition, the AO/EB stain result was indicated more death cells with belbing formation of the treated cells were observed. At 1000 µg/mL of IC50 concentration of chitosan loaded essential oils was damaged the A549 lung cancer cells with high necrotic cells, chromatin condensation, and decreased cell division. Therefore, all the invitro experiments results were clearly stated that the chitosan loaded essential oils as more efficient alternative option to inhibit the cancer cells.

A breakthrough in cosmeceuticals by utilizing insects as major ingredients in cosmetic products is gaining popularity. Therefore, the interest in rare sources of ingredients, for instance, from the Oryctes rhinoceros beetle, can bring huge benefits in terms of turning pests into wealth. In this study, curcumin was chosen as the active ingredient loaded into chitosan-gold nanoparticles (CCG-NP). Curcumin is unstable and has poor absorption, a high rate of metabolism, and high sensitivity to light. These are all factors that contribute to the low bioavailability of any substance to reach the target cells. Therefore, chitosan extracted from O. rhinoceros could be used as a drug carrier to overcome these limitations. In order to overcome these limitations, CCG-NPs were synthesized and characterized. Chitosan was isolated from O. rhinoceros and CCG-NPs were successfully synthesized at 70 °C for 60 min under optimal conditions of a reactant ratio of 2:0.5 (0.5 mM HAuCl4: 0.1% curcumin). Characterizations of CCG-NP involved FTIR analysis, zeta potential, morphological properties determination by FE-SEM, particle size analysis, crystallinity study by XRD, and elemental analysis by EDX. The shape of the CCG-NP was round, its size was 128.27 d.nm, and the value of the zeta potential was 20.2 ± 3.81 mV. The IC50 value for cell viability is 58%, indicating a mild toxicity trait. To conclude, CCG-NP is a stable, spherical, nano-sized, non-toxic, and homogeneous solution. [ABSTRACT FROM AUTHOR]

Background: This study aims to prepare high stability chitosan nanoparticles (CNP) and examine the ability of CNP in CpG-ODN delivery when treating allergic mice model. Methods: Preparation and characterization of CNP were performed by ionic gelation, dynamic light scattering, and zeta sizer. The CNP cytotoxicity and activation ability of CpG ODN delivered with CNP were tested using a cell counting kit-8 and Quanti blue method. Allergic mice were injected intraperitoneal with 10 ug ovalbumin on day 0 and 7, and then treated with intranasal CpG ODN/CpG ODN, delivered with CNP/CNP, on the third week three times per week for three weeks. The ELISA method measured cytokine and IgE profiles in the allergic mice's plasma and spleen. Results: CNP results have sizes 27.73 nm±3.67 dan 188.23 nm±53.47, spherical in shape and nontoxic, and did not alter the NF-κB activation of CpG ODN in RAW-blue cells. The application of CpG ODN delivered by chitosan nanoparticles shows no statistical difference between groups of IFN-κ, IL-10, and IL-13 in Balb/c mice's plasma and spleen, in contrast with IgE level. Conclusions: The results showed that using chitosan nanoparticles as a delivery system for CpG ODN has the potency to safely CpG ODN efficacy. [ABSTRACT FROM AUTHOR]

Onychomycosis is a nail infection caused by a fungus, Trichophyton mentagrophytes, that is responsible for major nail infections. The best method suited for treating such infections generally includes a topical remedy. However, conventional oral or topical formulations are associated with various limitations. Therefore, a more efficient and compatible formulation is developed in this study. The primary objective of the current study is to formulate and evaluate chitosan nanoparticle-based hydrogel for ameliorating onychomycosis. The sole purpose of this research was to increase the permeation of the lipophilic drug itraconazole and difluorinated curcumin, and its synergistic antifungal activity was also evaluated for the first time. Both in vitro and ex vivo drug release evaluations confirmed the sustained release of both drugs from the hydrogel, which is a prerequisite for treating onychomycosis. The results overall highlighted the promising activity of a synergistic approach that could be implemented for the treatment of onychomycosis. The hydrogel-based formulation serves as an effective method of delivery of drugs across the layers of the skin, resulting from its hydrating characteristics. [ABSTRACT FROM AUTHOR]

The controlled-release characteristic of drug delivery systems is utilized to increase the residence time of therapeutic agents in the human body. This study aimed to formulate and characterize salsalate (SSL)-loaded chitosan nanoparticles (CSNPs) prepared using the ionic gelation method and to assess their in vitro release and antibacterial and antibiofilm activities. The optimized CSNPs and CSNP–SSL formulation were characterized for particle size (156.4 ± 12.7 nm and 132.8 ± 17.4 nm), polydispersity index (0.489 ± 0.011 and 0.236 ± 132 0.021), zeta potential (68 ± 16 mV and 37 ± 11 mV), and entrapment efficiency (68.9 ± 2.14%). Physicochemical features of these nanoparticles were characterized using UV–visible and Fourier transform infrared spectroscopy and X-ray diffraction pattern. Scanning electron microscopy studies indicated that CSNPs and CSNP–SSL were spherical in shape with a smooth surface and their particle size ranged between 200 and 500 nm. In vitro release profiles of the optimized formulations showed an initial burst followed by slow and sustained drug release after 18 h (64.2 ± 3.2%) and 48 h (84.6 ± 4.23%), respectively. Additionally, the CSNPs and CSNP–SSL nanoparticles showed a sustained antibacterial action against Staphylococcus aureus (15.7 ± 0.1 and 19.1 ± 1.2 mm) and Escherichia coli (17.5 ± 0.8 and 21.6 ± 1.7 243 mm). Interestingly, CSNP–SSL showed better capability (89.4 ± 1.2% and 95.8 ± 0.7%) than did CSNPs in inhibiting antibiofilm production by Enterobacter tabaci (E2) and Klebsiella quasipneumoniae (SC3). Therefore, CSNPs are a promising dosage form for sustained drug delivery and enhanced antibacterial and antibiofilm activity of SSL; these results could be translated into increased patient compliance. [ABSTRACT FROM AUTHOR]

Recently, various innovative technologies have been developed for the enhanced delivery of biologics as attractive formulation targets including polymeric micro and nanoparticles. Combined with personalized medicine, this area can offer a great opportunity for the improvement of therapeutics efficiency and the treatment outcome. Herein, a novel manufacturing method has been introduced to produce protein-loaded chitosan particles with controlled size. This method is based on an additive manufacturing technology that allows for the designing and production of personalized particulate based therapeutic formulations with a precise control over the shape, size, and potentially the geometry. Sprayed multi adsorbed-droplet reposing technology (SMART) consists of the high-pressure extrusion of an ink with a well determined composition using a pneumatic 3D bioprinting approach and flash freezing the extrudate at the printing bed, optionally followed by freeze drying. In the present study, we attempted to manufacture trypsin-loaded chitosan particles using SMART. The ink and products were thoroughly characterized by dynamic light scattering, rheometer, Scanning Electron Microscopy (SEM), and Fourier Transform Infra-Red (FTIR) and Circular Dichroism (CD) spectroscopy. These characterizations confirmed the shape morphology as well as the protein integrity over the process. Further, the effect of various factors on the production were investigated. Our results showed that the concentration of the carrier, chitosan, and the lyoprotectant concentration as well as the extrusion pressure have a significant effect on the particle size. According to CD spectra, SMART ensured Trypsin's secondary structure remained intact regardless of the ink composition and pressure. However, our study revealed that the presence of 5% (w/v) lyoprotectant is essential to maintain the trypsin's proteolytic activity. This study demonstrates, for the first time, the viability of SMART as a single-step efficient process to produce biologics-based stable formulations with a precise control over the particulate morphology which can further be expanded across numerous therapeutic modalities including vaccines and cell/gene therapies. [ABSTRACT FROM AUTHOR]

The current study was conducted to evaluate the effect of supplementing fish and gluten meals with chitosan and chitosan nanoparticles on the Nile tilapia growth performance, feed utilization, fatty acids profile and liver histology. The experimental design was completely randomized with a 2 x 3 factorial design; two protein sources; fish and gluten meals, and three forms of chitosan (zero-chitosan, chitosan and chitosan nanoparticles). A total number of 270 O. niloticus fingerlings were randomly distributed among 18 tanks (55 liters water capacity). The experiment lasted for 82 days. Results indicated that the growth performance parameters and protein efficiency ratio of fish fed gluten meal-based diets were improved by the addition of chitosan nanoparticles. Additionally, it was found that supplementation of gluten meal-based diet with chitosan nanoparticles increased the total omega-3 level, while decreased the omega-6/omega-3 ratio. Moreover, supplementation of gluten meal-based diets with chitosan nanoparticles improved the histological architecture of the liver and reduced fish mortality. Consequently, it is recommended to use chitosan nanoparticles as feed additives to gluten meal-based diets at a level of 0.5%. [ABSTRACT FROM AUTHOR]

Background: Red ginseng (RG) extract, especially ginsenoside Rg1 and Rb1 fractions has been reported to have antithrombotic activities. However, gastric instability and low intestinal permeability are considered to be obstacles to its oral administration. We hypothesized that stability, permeability, and activities of RG might be improved by encapsulation within nanoparticles (NPs) prepared with antithrombotic coating materials. Methods: RG-loaded chitosan (CS) NPs (PF-NPs) were prepared by complex ionic gelation with the antithrombotic wall materials, polyglutamic acid (PGA), and fucoidan (Fu). The concentrations of PGA (mg/mL, X1) and Fu (mg/mL, X2) were optimized for the smallest particle size by response surface methodology. Antithrombotic activities of RG and PF-NPs were analyzed using ex vivo and in vivo antiplatelet activities, in vivo carrageenan-induced mouse tail, and arteriovenous shunt rat thrombosis models. Results: In accordance with a quadratic regression model, the smallest PF-NPs (286 ± 36.6 nm) were fabricated at 0.628 mg/mL PGA and 0.081 mg/mL Fu. The inhibitory activities of RG on ex vivo and in vivo platelet aggregation and thrombosis in in vivo arteriovenous shunt significantly (p

Introduction: Methamphetamine (MA) acts as a powerful oxidant agent, while Rosmarinic Acid (RA) is an effective herbal antioxidant. Oxidative stress-mediated by MA results in apoptosis, and caspase-3 is one of the critical enzymes in the apoptosis process. MA can epigenetically alter gene regulation. In this paper, to investigate the effects of RA on MA-mediated oxidative stress, changes in the level of casp3a mRNA were demonstrated in zebrafish. Methods: The animals were grouped in 3 treatment conditions for the behavioral test: control, MA, MA pretreated by RA, and 6 treatment conditions for the molecular test: control, RA, MA, MA co-treated with RA, MA co-treated with RA/ZnO/chitosan nanoparticle, and ZnO/chitosan nanoparticle. Then molecular and behavioral investigations were carried out, and critical comparisons were made between the groups. MA solution was prepared with a concentration of 25 mg/L, and RA solution was prepared by DPPH test with the antioxidant power of about 97%. Each solution was administered by immersing 20 zebrafish for 20 minutes, once per day for 7 days. The level of casp3a mRNA was quantified by using qRT-PCR. One-sided trapezoidal tank diving test was applied to study behavioral alterations. Results: The qPCR analysis demonstrated the high potential of RA/ZnO/chitosan in counteracting the MA-mediated elevation in casp3a mRNA level. Based on the diving test results of MA-treated fish, MA was found to be anxiolytic compared to the control. While the resulted diving pattern of the MA-treated animals pretreated by RA was novel and different from both the control and MA-treated groups. Conclusion: The potential of RA combined with a suitable nanoparticle against MA-induced oxidative stress was supported. The high efficiency of ZnO/chitosan in increasing RA penetration to the brain cells was evident. MA at a dose of 25 mg/L is anxiolytic for zebrafish. However, the molecular mechanisms involved in these processes should be studied.

The Newcastle disease virus (NDV) is considered a serious threat to global poultry production. Despite the availability of vaccines, it remains a major devastating epidemic responsible for great economic losses. The development of novel virus-controlling strategies is therefore an urgent need. The present study investigated for the first time the antiviral efficacy of propolis and chitosan nanoparticles against two NDV isolates, MW881875 and MW881876, recovered from vaccinated commercial broiler farms in KafrEl Sheikh Governorate, Egypt. The polygenetic analysis focused on the F and M genes, with one isolate having a 97% identity with the genotype VII NDV Israeli strain. On the other hand, the identified isolates showed high genetic variation and only 76% identity with the LaSota vaccine (genotype II). More interestingly, the cell cytotoxic concentrations of chitosan, propolis, and a propolis–chitosan mixture against Vero cells were 327.41 ± 12.63, 109.48 ± 8.36, and 231.78 ± 11.46 μg/ml, respectively. The median tissue culture infectious dose (TCID50) assay demonstrated that the nanoparticles have antiviral effects after NDV exposure resulting in significant decrease in viral titer (TCID50) by 2, 2.66, and 2.5 log10 at 62 μg/ml of chitosan, 13 μg/ml of propolis, and 30 μg/ml of the propolis–chitosan mixture, respectively, compared with the control TCID50 value of 4 log10. Taken together, the results provide novel insights into the potentially promising roles of propolis and chitosan as novel, safe, and effective antiviral agents against NDV.

Hepatocellular carcinoma (HCC) is one of the major causes of cancer-related mortality worldwide. Nowadays, liver-targeting drug delivery system has been proven as a promising strategy for overcoming HCC. Asialoglycoprotein receptor (ASGPR) is an ideal receptor for liver targeting, which is mainly expressed on hepatocytes. In this study, we developed several novel liver-targeting chitosan nanoparticles to selectively overcome HCC via ASGPR. Chitosan nanoparticles (Gly-CS-VE, Gal-Gly-CS-VE, Gly-CS-DCA, and Gal-Gly-CS-DCA) were prepared by grafting hydrophilic group (glycidol, Gly), hydrophobic group (deoxycholic acid, DCA or vitamin E succinate, VE), and ASGPR recognizing group (galactose, Gal). Subsequently, their characterizations were measured by 1H NMR, FT-IR, TEM, and DLS. Doxorubicin (DOX) was loaded in nanoparticles and released out in a pH-dependent manner. Most importantly, the galactosylated Gal-Gly-CS-VE and Gal-Gly-CS-DCA nanoparticles exhibited significantly stronger in vitro cell internalization, cytotoxicity, anti-migration capabilities and in vivo anticancer efficacies than the corresponding Gly-CS-VE and Gly-CS-DCA nanoparticles, as well as free DOX. Finally, the four chitosan nanoparticles exhibited good biocompatibility without causing any obvious histological damage to the major organs. Overall, the galactosylated chitosan nanoparticles were proven to be promising pharmaceutical formulations for selectively overcoming HCC, with great potential for clinical applications.

Toxoid vaccines can provide protective immunity against clostridial diseases. Since the duration of the toxoid vaccine immunogenicity is short, these vaccines need to contain an adjuvant. The nanoparticles of chitosan can stimulate humoral and cell-mediated immune responses. In the present study, the effect of chitosan nanoparticles was investigated on the immunogenicity of the pentavalent clostridial toxoid vaccine containing Clostridium perfringens types D, C, and B, Clostridium septicum, as well as Clostridium novyi. Rabbits were immunized by two injections with 3-week intervals and checked clinically and through autopsy 2 weeks after the last injection. Hematological changes were investigated during immunization, including the changes of white and red blood cell counts, hemoglobin, packed cell volume, platelet, neutrophil, lymphocyte, eosinophil, basophile, monocyte, and Neut/Lymph. Biochemical factors, namely creatinine, blood urea nitrogen, glucose, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total protein, and albumin, were also studied. The changes in immune responses during the immunization period were investigated through indirect enzyme-linked immunosorbent assay (ELISA). The results of ELISA showed that chitosan significantly enhanced immunogenicity when accompanied with in the pentavalent clostridial toxoid vaccine. During the immunogenicity period and following that, no changes were observed in clinical behavior and internal organs after autopsy. The hematological and biochemical factors were reported with no significant pathologic changes during immunization in the control and vaccinated groups (P

Masoumeh Saberpour, Bita Bakhshi, Shahin Najar-peerayeh Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University Tehran, Tehran, IranCorrespondence: Bita BakhshiDepartment of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Jalal-Ale-Ahmad Ave, Tehran 14117-13116, IranTel/ Fax +982182884558Email b.bakhshi@modares.ac.irAim: The aim of the present study was to evaluate the in vitro antimicrobial and antibiofilm activity of chitosan nanoparticles (CS NPs) incorporated with mesenchymal stem cells-derived conditioned media (MSCs CM) on MDR Vibrio cholerae strains.Materials and Methods: Chitosan NPs were prepared and characterized by dynamic light scattering (DLS), scanning electron microscope (SEM) and zeta potential measurement. MSCs CM were prepared and entrapped into MSCs CM-CS NPs composite and its release efficiency was measured. Antibacterial efficacy of nano structures was determined by disk diffusion and broth microdilution methods. Antibiofilm activity was assessed by crystal violet assay.Results: BM-MSCs were characterized to be negative for CD34 and CD45 markers, positive for CD73 and CD44 markers, and able to differentiate into osteoblast and adipocyte cells. The mean particle size of 96.6% of chitosan NPs was 414.9 nm with a suitable zeta potential and SEM morphology. Entrapment efficiency of MSCs CM-CS NPs was 76.9%. Unstimulated MSCs CM-CS NPs composite as a novel and proficient therapeutic nanostructure against MDR V. cholerae strains showed the synergistic activity of the two components of MSCs CM and CS NPs, leading to greater bacterial killing compared to control groups. MSCs CM more efficiently inhibited biofilm formation, although MSCs CM-CS NPs was also appeared to be effective in inhibiting biofilm formation compared to CS NPs and control group.Conclusion: The designed nanodrug composite showed the best release in conditions mimicking the physiological conditions of the intestinal lumen. Given the fact that no overuse or genetic event would cause the emergence of antimicrobial resistance against the MSCs CM-CS NPs nanodrug, it could be considered as a promising alternative for the treatment of MDR V. cholerae infections in clinical settings.Keywords: Vibrio cholerae, antimicrobial, chitosan nanoparticle, mesenchymal stem cells

Recently, the research and innovation to produce raw materials from microbial processes has gained much attention due to their economic and environmental impacts. Lactic acid is a very important microbial product due to its wide application in the food, pharmaceutical, cosmetic, and chemical industries. In the current study, poly (L-lactic acid) (PLLA) was produced by the ring opening polymerization (ROP) technique of L-lactic acid recovered from whey fermentation, and was used for the production of nanocomposites films reinforced with chitosan nanoparticles (CNPs) (average diameter ca. 100–200 nm). Three different CNPs concentrations, namely 1, 3, and 5% w/w, were tested, and their influence on the physical, mechanical, thermal, antibacterial and structural attributes of PLLA film was assessed. The results showed that the addition of CNPs up to 3% caused a significant improvement in water vapor permeability, appearance, tensile strength and elongation at break. The antibacterial properties of nanocomposites followed a dose-depended pattern as a result of CNPs addition. Therefore, the best inhibitory effects on Escherichia coli and Staphylococcus aureus was made by the addition of 5% of CNPs and lower dosages slightly affected the growth of pathogens or didn't cause any inhibitory effects (in 1% of CNPs). It can be concluded that the incorporation of CNPs into the PLLA matrix allows to improve the structural, thermal, physical, mechanical and antibacterial properties of the polymer, generating promising systems for food packaging and biomedical applications.

Aquaculture aims to provide stable environmental conditions for fish to be able to grow and protect it from stress and diseases. In order to guarantee a sustainable yield of fish species, well-balanced diets should be considered. For this purpose, diets supplemented with feed additives have won the interest of researchers who are concerned with the aquaculture field, the most important of which are chitosan and chitosan nanoparticles. Chitosan [β-(1-4)-N-acetyl-D-glucosamine] is a cationic biopolymer, derived from the alkaline deacetylation of chitin; a natural polymer found in the exoskeletons of insects, crustaceans and fungal cell walls. Chitosan nanoparticles were synthesized either by chemical or physical cross linking. Chitosan nanoparticles are favored due to their large surface area, bioavailability and deep penetration to the target sites. Chitosan and chitosan nanoparticles have several unique biological properties that enable them to act as promising materials in variable applications, including biosafety, biocompatibility, improved solubility and biodegradability. The afore-mentioned properties enabled them to be used in several applications in fish aquaculture, such as enhancing growth performance and immune functions. Moreover, they act as safe feed additives, drug carriers and are widely used in water treatment as well. The applications of chitosan and chitosan nanoparticles in fish aquaculture attracted the interest of many researchers. The present review covered all information concerning the effect of chitosan and chitosan nanoparticles on fish, including antioxidant activity, growth enhancement and immunostimulatory effects. The discussed studies in the current review proved that chitosan and chitosan nanoparticles are regarded as promising materials with wide applications in fish aquaculture. [ABSTRACT FROM AUTHOR]

Sankar Renu, 1 Ashley D Markazi, 2 Santosh Dhakal, 1 Yashavanth S Lakshmanappa, 1 Revathi Shanmugasundaram, 2 Ramesh K Selvaraj, 3 Gourapura J Renukaradhya 1 1Food Animal Health Research Program, Ohio Agricultural Research and Development Center, Wooster, OH 44691, USA and Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA; 2Department of Animal Sciences, Ohio Agricultural Research and Development Center, The Ohio State University, Columbus, OH, USA; 3Department of Poultry Science, University of Georgia, Athens, GA 30602, USACorrespondence: Gourapura J RenukaradhyaFood Animal Health Research Program, Ohio Agricultural Research and Development Center, The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691, USATel +1 330-263-3748Fax +1 330-263-3677Email gourapura.1@osu.eduRamesh K SelvarajDepartment of Poultry Science, University of Georgia, Athens, GA 30602, USAEmail selvaraj@uga.eduPurpose: Salmonellosis in poultry is a serious economic burden. A major concern is the public health hazard caused by consumption of Salmonella-contaminated poultry products. Currently used Salmonella vaccines are ineffective in combating poultry Salmonellosis warranting the need of a potent vaccine, especially an oral vaccine that can elicit robust local intestinal immunity.Materials and Methods: A Salmonella subunit chitosan nanoparticles (NPs)-based vaccine was prepared that contained immunogenic outer membrane proteins (OMPs) and -flagellin (F) protein (OMPs-F-CS NPs). OMPs-F-CS NPs were administered as an oral vaccine in layer chickens and the resultant humoral and cell-mediated immune responses and localization of NPs were examined using standard detection methods.Results: We demonstrated targeting of surface F-protein coated chitosan NPs to immune cells when delivered orally to layer chickens, the particles were localized in ileal Peyer’s patches. The OMPs-F-CS NPs vaccinated layer chickens had significantly higher OMPs-specific mucosal IgA production and lymphocyte proliferation response. The candidate vaccine increased the expression of toll-like receptor (TLR)-2, TLR-4, IFN-γ, TGF-ß and IL-4 mRNA expression in chicken cecal tonsils.Conclusion: Our study demonstrated that the chitosan-based oral Salmonella nanovaccine targets immune cells of chickens and induced antigen-specific B and T cell responses. This candidate oral Salmonella nanovaccine has the potential to mitigate Salmonellosis in poultry.Keywords: chickens, chitosan nanoparticle, Salmonella antigens, oral delivery, mucosal immune response

Li Lin,1,2,* Junyi He,2,* Jiawei Li,2 Yan Xu,2 Jingxia Li,2 Yangzhe Wu1,2 1Zhuhai Precision Medical Center, Zhuhai People’s Hospital, Zhuhai Hospital Affiliated with Jinan University, Jinan University, Zhuhai, Guangdong 519000, People’s Republic of China; 2The Biomedical Translational Research Institute, Faculty of Medical Science, Jinan University, Guangzhou, Guangdong 510632, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yangzhe WuZhuhai Precision Medical Center, Zhuhai People’s Hospital, Zhuhai Hospital Affiliated with Jinan University, Jinan University, Zhuhai, Guangdong 519000, People’s Republic of ChinaTel +86 020 8522 2787Email tyzwu@jnu.edu.cnBackground: During the past few years, immune cell therapy for malignant cancer has benefited a considerable amount of patients worldwide. As one of several promising candidates for immunotherapy, Vγ9Vδ2 γδ T cells have many unique biological advantages, such as non-MHC restriction and have been noted as the earliest source of IFN-γ. However, potentiating anti-tumor functions of γδ T cells has become of particular interest to researchers studying γδ T cell applications.Purpose: In this study, we proposed a nanotechnology-based methodology for strengthening γδ T cell functions.Methods: As a type of reliable, biocompatible material, chitosan nanoparticles (CSNPs) were used to enhance anti-tumor immunity of γδ T cells.Results: First, we found that the size of prepared CSNPs distributed 50 to 100 nm, and that CSNPs had optimal immunocompatibility. Then, we observed that CSNPs could induce α-tubulin cytoskeleton polarization and rearrangement, correlating with a higher killing ability of γδ T cells. Furthermore, we revealed that CSNPs could enhance Vγ9Vδ2 T cell anti-tumor functions by upregulating killing of related receptors, including NKG2D, CD56, FasL, and perforin secretion.Conclusion: Our work provided evidence of application for CSNPs based bio-carrier in immunotherapy. More importantly, we proposed a new strategy for enhancing γδ T cell anti-tumor activity using nanobiomaterial, which could benefit future clinical applications of γδ T cells.Keywords: chitosan nanoparticles, Vγ9Vδ2 γδ T cell, cytotoxicity, anti-tumor activity

The controlled-release characteristic of drug delivery systems is utilized to increase the residence time of therapeutic agents in the human body. This study aimed to formulate and characterize salsalate (SSL)-loaded chitosan nanoparticles (CSNPs) prepared using the ionic gelation method and to assess their in vitro release and antibacterial and antibiofilm activities. The optimized CSNPs and CSNP–SSL formulation were characterized for particle size (156.4 ± 12.7 nm and 132.8 ± 17.4 nm), polydispersity index (0.489 ± 0.011 and 0.236 ± 132 0.021), zeta potential (68 ± 16 mV and 37 ± 11 mV), and entrapment efficiency (68.9 ± 2.14%). Physicochemical features of these nanoparticles were characterized using UV–visible and Fourier transform infrared spectroscopy and X-ray diffraction pattern. Scanning electron microscopy studies indicated that CSNPs and CSNP–SSL were spherical in shape with a smooth surface and their particle size ranged between 200 and 500 nm. In vitro release profiles of the optimized formulations showed an initial burst followed by slow and sustained drug release after 18 h (64.2 ± 3.2%) and 48 h (84.6 ± 4.23%), respectively. Additionally, the CSNPs and CSNP–SSL nanoparticles showed a sustained antibacterial action against Staphylococcus aureus (15.7 ± 0.1 and 19.1 ± 1.2 mm) and Escherichia coli (17.5 ± 0.8 and 21.6 ± 1.7 243 mm). Interestingly, CSNP–SSL showed better capability (89.4 ± 1.2% and 95.8 ± 0.7%) than did CSNPs in inhibiting antibiofilm production by Enterobacter tabaci (E2) and Klebsiella quasipneumoniae (SC3). Therefore, CSNPs are a promising dosage form for sustained drug delivery and enhanced antibacterial and antibiofilm activity of SSL; these results could be translated into increased patient compliance.

Parenteral administration of killed/inactivated swine influenza A virus (SwIAV) vaccine in weaned piglets provides variable levels of immunity due to the presence of preexisting virus specific maternal derived antibodies (MDA). To overcome the effect of MDA on SwIAV vaccine in piglets, we developed an intranasal deliverable killed SwIAV antigen (KAg) encapsulated chitosan nanoparticles called chitosan-based NPs encapsulating KAg (CS NPs-KAg) vaccine. Further, to target the candidate vaccine to dendritic cells and macrophages which express mannose receptor, we conjugated mannose to chitosan (mCS) and formulated KAg encapsulated mCS nanoparticles called mannosylated chitosan-based NPs encapsulating KAg (mCS NPs-KAg) vaccine. In MDA-positive piglets, prime-boost intranasal inoculation of mCS NPs-KAg vaccine elicited enhanced homologous (H1N2-OH10), heterologous (H1N1-OH7), and heterosubtypic (H3N2-OH4) influenza virus-specific secretory IgA (sIgA) antibody response in nasal passage compared to CS NPs-KAg vaccinates. In vaccinated upon challenged with a heterologous SwIAV H1N1, both mCS NPs-KAg and CS NPs-KAg vaccinates augmented H1N2-OH10, H1N1-OH7, and H3N2-OH4 virus-specific sIgA antibody responses in nasal swab, lung lysate, and bronchoalveolar lavage (BAL) fluid; and IgG antibody levels in lung lysate and BAL fluid samples. Whereas, the multivalent commercial inactivated SwIAV vaccine delivered intramuscularly increased serum IgG antibody response. In mCS NPs-KAg and CS NPs-KAg vaccinates increased H1N2-OH10 but not H1N1-OH7 and H3N2-OH4-specific serum hemagglutination inhibition titers were observed. Additionally, mCS NPs-KAg vaccine increased specific recall lymphocyte proliferation and cytokines IL-4, IL-10, and IFNγ gene expression compared to CS NPs-KAg and commercial SwIAV vaccinates in tracheobronchial lymph nodes. Consistent with the immune response both mCS NPs-KAg and CS NPs-KAg vaccinates cleared the challenge H1N1-OH7 virus load in upper and lower respiratory tract more efficiently when compared to commercial vaccine. The virus clearance was associated with reduced gross lung lesions. Overall, mCS NP-KAg vaccine intranasal immunization in MDA-positive pigs induced a robust cross-reactive immunity and offered protection against influenza virus.

Parenteral administration of killed/inactivated swine influenza A virus (SwIAV) vaccine in weaned piglets provides variable levels of immunity due to the presence of preexisting virus specific maternal derived antibodies (MDA). To overcome the effect of MDA on SwIAV vaccine in piglets, we developed an intranasal deliverable killed SwIAV antigen (KAg) encapsulated chitosan nanoparticles called chitosan-based NPs encapsulating KAg (CS NPs-KAg) vaccine. Further, to target the candidate vaccine to dendritic cells and macrophages which express mannose receptor, we conjugated mannose to chitosan (mCS) and formulated KAg encapsulated mCS nanoparticles called mannosylated chitosan-based NPs encapsulating KAg (mCS NPs-KAg) vaccine. In MDA-positive piglets, prime-boost intranasal inoculation of mCS NPs-KAg vaccine elicited enhanced homologous (H1N2-OH10), heterologous (H1N1-OH7), and heterosubtypic (H3N2-OH4) influenza virus-specific secretory IgA (sIgA) antibody response in nasal passage compared to CS NPs-KAg vaccinates. In vaccinated upon challenged with a heterologous SwIAV H1N1, both mCS NPs-KAg and CS NPs-KAg vaccinates augmented H1N2-OH10, H1N1-OH7, and H3N2-OH4 virus-specific sIgA antibody responses in nasal swab, lung lysate, and bronchoalveolar lavage (BAL) fluid; and IgG antibody levels in lung lysate and BAL fluid samples. Whereas, the multivalent commercial inactivated SwIAV vaccine delivered intramuscularly increased serum IgG antibody response. In mCS NPs-KAg and CS NPs-KAg vaccinates increased H1N2-OH10 but not H1N1-OH7 and H3N2-OH4-specific serum hemagglutination inhibition titers were observed. Additionally, mCS NPs-KAg vaccine increased specific recall lymphocyte proliferation and cytokines IL-4, IL-10, and IFNγ gene expression compared to CS NPs-KAg and commercial SwIAV vaccinates in tracheobronchial lymph nodes. Consistent with the immune response both mCS NPs-KAg and CS NPs-KAg vaccinates cleared the challenge H1N1-OH7 virus load in upper and lower respiratory tract more efficiently when compared to commercial vaccine. The virus clearance was associated with reduced gross lung lesions. Overall, mCS NP-KAg vaccine intranasal immunization in MDA-positive pigs induced a robust cross-reactive immunity and offered protection against influenza virus. [ABSTRACT FROM AUTHOR]

Introduction: Methamphetamine (MA) acts as a powerful oxidant agent, while Rosmarinic Acid (RA) is an effective herbal antioxidant. Oxidative stress-mediated by MA results in apoptosis, and caspase-3 is one of the critical enzymes in the apoptosis process. MA can epigenetically alter gene regulation. In this paper, to investigate the effects of RA on MA-mediated oxidative stress, changes in the level of casp3a mRNA were demonstrated in zebrafish. Methods: The animals were grouped in 3 treatment conditions for the behavioral test: control, MA, MA pretreated by RA, and 6 treatment conditions for the molecular test: control, RA, MA, MA co-treated with RA, MA co-treated with RA/ZnO/chitosan nanoparticle, and ZnO/chitosan nanoparticle. Then molecular and behavioral investigations were carried out, and critical comparisons were made between the groups. MA solution was prepared with a concentration of 25 mg/L, and RA solution was prepared by DPPH test with the antioxidant power of about 97%. Each solution was administered by immersing 20 zebrafish for 20 minutes, once per day for 7 days. The level of casp3a mRNA was quantified by using qRT-PCR. One-sided trapezoidal tank diving test was applied to study behavioral alterations. Results: The qPCR analysis demonstrated the high potential of RA/ZnO/chitosan in counteracting the MA-mediated elevation in casp3a mRNA level. Based on the diving test results of MA-treated fish, MA was found to be anxiolytic compared to the control. While the resulted diving pattern of the MA-treated animals pretreated by RA was novel and different from both the control and MA-treated groups. Conclusion: The potential of RA combined with a suitable nanoparticle against MA-induced oxidative stress was supported. The high efficiency of ZnO/chitosan in increasing RA penetration to the brain cells was evident. MA at a dose of 25 mg/L is anxiolytic for zebrafish. However, the molecular mechanisms involved in these processes should be studied. [ABSTRACT FROM AUTHOR]

Owing to their nanoscale dimensions, nanomaterials have special chemical and physical properties that set them apart from their bulk counterparts. The exterior dimensions of a minimum of half of the particles span several nanometers in their size distribution. Silver nanoparticles (AgNPs) are one type of nanomaterial that has been widely used because of their strong antibacterial properties, which can kill bacteria that are resistant to many drugs. Due to its potential for regulated release, localized retention, and safeguarding the active ingredients against environmental or enzymatic deterioration, nanoparticle technology has also emerged as a promising medication delivery method. The techniques for creating nanoparticles can be easily scaled up and used for a wide variety of medications. Since polymeric nanoparticles are biodegradable, biocompatible, and have more readily available formulation techniques than other nanoparticle drug delivery approaches, their range of applications has been expanding. Chitosan, also known as deacetylated polysaccharide, is a straight-chain cationic polymer that is typically a cationic copolymer. It can be generated naturally or by deacetylating chitin. Consequently, it contains an extensive array of biomedical applications, such as efficient healing of wounds, regeneration of tissues, regeneration of bone, and anti-infection. Because of its functional diversity, accessibility, and being both biodegradable and biocompatible, it has a wide spectrum of uses in dentistry. Recent research on chitosan-based nanoparticles is founded on the field's growing comprehension of the characteristics of chitosan and techniques for chemical or physical modification that are used to optimize the drug loading and release characteristics of the nanoparticles., Competing Interests: Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Thakare et al.)

The cytokine storm or cytokine storm syndrome (CSS) is associated with high mortality in patients with acute lung injury (ALI) and acute respiratory distress syndrome (ARDS), for example following sepsis or infectious diseases including COVID-19. However, there are no effective treatments for CSS-associated ALI or ALI/ARDS. Thus, there remains an urgent need to develop effective drugs and therapeutic strategies against CSS and ALI/ARDS. Nasal and inhaled drug delivery methods represent a promising strategy in the treatment of inflammatory lung disease as a result of their ability to improve drug delivery to lungs. Improving the nasal mucosa absorption of poorly water-soluble drugs with poor mucosa bioavailability to a therapeutically effective level is another promising strategy in the fight against ALI/ARDS. Here, chitosan nanoparticles loaded with hesperidin (HPD/NPs) were developed for nasal delivery of the anti-inflammatory HPD compound to inflammatory lungs. In vitro and in vivo, HPD/NPs exhibited enhanced cellular uptake in the inflammatory microenvironment compared with free HPD. In a mouse model of inflammatory lung disease, the HPD/NPs markedly inhibited lung injury as evidenced by reduced inflammatory cytokine levels and suppressed vascular permeability compared with free HPD. Collectively, our study demonstrates that nasal delivery of HPD/NPs suppresses CSS and ALI/ARDS in a murine model of inflammatory lung disease, and that nanoparticle-based treatment strategies with anti-inflammatory effects could be used to reduce CSS and ALI in patients with inflammatory lung injury.

Infectious disease of poultry and pig are major threat to health and cause severe economic loss to the food industry and a global food safety issue. Poultry and pig act as a mixing vessel of zoonotic transmission of disease to humans. Effective mucosal vaccines used in animals could reduce the impact of diseases in food animals. Chitosan is a biocompatible polymer, and its positive charge makes it a natural mucoadhesive agent. Therefore, since last one-decade chitosan derived nanoparticles (CS NPs) have been in use widely to deliver vaccine antigens in animals through mucosal route. Primary route of entry of most infectious disease pathogen is through oral and nasal routes, and the CS NPs based vaccines delivered through that routes enhance the immunogenicity of encapsulated vaccine antigens by targeting the cargo to mucosal microfold cells, dendritic cells and macrophages. Resulting in induction of robust secretory and systemic antibodies and/or cell mediated immune response which provides protection against infections. To date, CS NPs is being widely used for mucosal vaccine delivery in poultry and pigs to control bacterial and viral infections, and tested in several preclinical trials for vaccine delivery in humans. In this review, we highlighted the progress so far made in using CS NPs as a vehicle for mucosal vaccine delivery against infectious and zoonotic diseases of poultry and pigs. Discussed about the need of CS NPs modifications, CS NPs based vaccines induced immune responses and its role in protection, and challenges in vaccination and future directions.

Abstract Acne Vulgaris is a common skin disease caused by Propionibacterium acnes, an anaerobic microbiota of human skin that plays a vital role in the pathology of acne. The aim of this study was to prepare nanoparticles containing an acne recombinant protein and determine its ability as an oral acne vaccine in mice. The recombinant Sialidase-CAMP gene was expressed and purified in a prokaryotic host. The chitosan nanoparticles containing the recombinant protein were prepared, encapsulated, and administered by both oral and subcutaneous routes to Balb/c mice. Sera IgA and IgG and stool IgA titers were measured by ELISA, and the immunized mice were challenged against P. acnes. A 65 kDa recombinant protein was confirmed by SDS-PAGE and western blot. The size and zeta potential of nanoparticles were 80 nm and +18 mV, respectively. After oral immunization, the serum IgG and IgA titers were 1:3200 and 1:16, respectively, and the stool IgA titer was 1:8. In the subcutaneous route, the serum IgG titer was 1:51200. Immunized mice showed no inflammation in the ear of challenged mice. It is the first study that examines a chitosan-nanoparticulated acne fusion protein as an applicable acne vaccine candidate with appropriate immunogenicity potential. Further studies are required to validate the clinical usefulness of this vaccine candidate.

Most infectious diseases are caused by pathogenic infiltrations from the mucosal tract. Nowadays, the use of vaccines has been widely investigated for the prevention of different infectious diseases, infertility, immune disorders, malignancies, and allergies. Broad-spectrum adjuvant substances have been studied for immune system stimulation with a greater efficiency against specific antigens. Various adjuvants have been developed such as inorganic, oil-based, and emulsion adjuvants, bacterial products and their derivatives, cytokines, cytosine-guanine dinucleotide (CpG) motifs, and particulate systems. Mucosal vaccine delivery is an alternative route to induce both humoral and cellular immune responses. Applying nanoparticles in vaccine formulations allows not only improved antigen stability and immunogenicity, but also targeted delivery, and consequently, more specific release of the agent of interest. Chitosan nanoparticles have immunological activity and mucoadhesive properties. They have been used as a mucosal vaccine delivery system for many antigens. This review provides an overview of the recent advances in chitosan nanoparticles as a novel mucosal vaccine delivery system.

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An asymmetric resorbable‐based guided bone regeneration barrier membrane involved a loose layer of icariin@chitosan nanoparticles deposited on electrospun nanofibers, followed by a layer of tetracycline-loaded chitosan and gelatin-alginate hydrogel as the dense layer. In addition to enhancing osteogenic activity, the membrane's asymmetric design protects the bone defect from invasion of surrounding connective tissue and bacterial infection. [Display omitted] • An asymmetric membrane was designed to achieve targeted dual release for guided bone regeneration. • Icarin-loaded fibers and chitosan/tetracycline hydrogel were used for assembly of the membrane. • Long-lasting growth inhibition against S. aureus was achieved through hydrogel dense layer. • Fostering osteogenic differentiation was observed from icariin-loaded loose layer. Presently, guided bone regeneration (GBR) stands as a preeminent strategy for rectifying periodontal and maxillofacial bone imperfections. Nonetheless, the current GBR membranes encounter limitations in terms of antibacterial efficacy and osteogenic potential, impeding their overall effectiveness. To address this, our study endeavors to engineer an innovative GBR membrane with dual attributes of osteoinductiveness and antibacterial activity. This novel asymmetric nanocomposite integrates an intricate structure comprising a compact chitosan hydrogel layer embedded with tetracycline (TCH) and a porous polycaprolactone fibrous layer incorporating chitosan nanoparticles loaded with icariin (IcA@CNps). In the initial phase, IcA@CNps, exhibiting a balanced CNps to IcA ratio of 1:2, an average dimension of 80.94 ± 33.78 nm, and a zeta potential of −19.3 mV, were synthesized and affixed onto nanofibers. This integration facilitated the controlled release of icariin over a span of 22 days. Subsequently, the asymmetric nanocomposite GBR membrane (referred to as GBR +) was meticulously crafted. This involved the application of a TCH hydrogel laden with a drug concentration of 120 µg/ml onto the IcA@CNps-coated nanofibrous layer through an intermediary hydrogel stratum. Impressive results were witnessed during in vitro trials against Staphylococcus aureus , showcasing the remarkable antibacterial prowess of the GBR + membrane. Notably, human alveolar bone marrow stem cells (aBMSCs) cultivated on the GBR + membrane exhibited favorable cell adhesion and proliferation rates, devoid of any cytotoxic manifestations. Gene expression analyses further unveiled elevated expressions of SPP1, BMP2 , and RUNX2 in the proximity of the fabricated membrane, indicating the promotion of bone formation pathways. Consequently, this asymmetric GBR membrane presents a promising therapeutic avenue for bolstering bone regeneration, underscored by its dual action of fostering bone growth while concurrently displaying antibacterial attributes. [ABSTRACT FROM AUTHOR]

A mixed culture (polymicrobial) biofilm provides a favorable environment for pathogens to persist in the food processing environment and to contaminate food products. Inactivation and eradication of such biofilms from food processing environments are achieved by using harsh disinfectants, but their toxicity and environmentally hostile characteristics are unsustainable. This study aims to use food-grade natural nanoparticulated antimicrobials to control mixed-culture biofilms. Chitosan, a natural broad-spectrum antimicrobial biopolymer (polysaccharide) from crustaceans, was derivatized to produce chitosan nanoparticles (ChNP) as a carrier for another broad-spectrum antimicrobial agent, ε-poly-L-lysine (PL), to synthesize ChNP-PL conjugate. The antimicrobial activity of ChNP and ChNP-PL was tested against mixed-culture biofilms. ChNP-PL (~100 nm) exhibited a synergistic antimicrobial and anti-biofilm effect against mono or mixed-culture biofilms of five foodborne pathogens, including Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica serovar Enteritidis, Escherichia coli O157:H7, and Pseudomonas aeruginosa. ChNP-PL treatment prevented biofilm formation by mono or mixed cultures of L. monocytogenes, P. aeruginosa, and E. coli O157:H7, and bacterial counts were either below the detection limit or caused 3.5–5 log reduction. ChNP-PL also inactivated preformed biofilms. In monoculture biofilm, ChNP-PL treatment reduced L. monocytogenes counts by 4.5 logs, S. Enteritidis by 2 logs, E. coli by 2 logs, and S. aureus by 0.5 logs, while ChNP-PL had no inhibitory effect on P. aeruginosa. In vitro mammalian cell-based cytotoxicity analysis confirmed ChNP-PL to have no deleterious effect on intestinal HCT-8 cell line. In conclusion, our results show ChNP-PL has strong potential to prevent the formation or inactivation of preformed polymicrobial biofilms of foodborne pathogens.

The present study aimed to investigate the effects of chitosan (CS)-tripolyphosphate (TPP) nanoparticles (NPs) on the stability, antioxidant activity, and bioavailability of astaxanthin (ASX). ASX-loaded CS-TPP NPs (ACT-NPs) prepared by ionic gelation between CS (0.571 mg/mL) and TPP (0.571 mg/mL) showed 505.2 ± 184.8 nm, 20.4 ± 1.2 mV, 0.348 ± 0.044, and 63.9 ± 3.0% of particle size, zeta potential, polydispersity index and encapsulation efficiency, respectively. An in vitro release study confirmed that the release of ASX in simulated gastric (pH 1.2) and intestinal (pH 6.8) fluid was prolonged within ACT-NPs. The in vitro antioxidant activities of ACT-NPs were significantly improved compared with free ASX (FA) (p < 0.05). Furthermore, the cellular and in vivo antioxidant analysis verified that ACT-NPs could enhance the cytoprotective effects on the BHK-21 cell line and demonstrate sustained release properties, leading to prolonged residence time in the rat plasma. The results suggest that the stability, antioxidant properties, and bioavailability of ASX can be effectively enhanced through encapsulation within CS-TPP NPs.

Lung cancer has been recognized as one of the most often diagnosed and perhaps most lethal cancer diseases worldwide. Conventional chemotherapy for lung cancer-related diseases has bumped into various limitations and challenges, including non-targeted drug delivery, short drug retention period, low therapeutic efficacy, and multidrug resistance (MDR). Chitosan (CS), a natural polymer derived from deacetylation of chitin, and comprised of arbitrarily distributed β-(1-4)-linked d-glucosamine (deacetylated unit) and N-acetyl-d-glucosamine (acetylated unit) that exhibits magnificent characteristics, including being mucoadhesive, biodegradable, and biocompatible, has emerged as an essential element for the development of a nano-particulate delivery vehicle. Additionally, the flexibility of CS structure due to the free protonable amino groups in the CS backbone has made it easy for the modification and functionalization of CS to be developed into a nanoparticle system with high adaptability in lung cancer treatment. In this review, the current state of chitosan nanoparticle (CNP) systems, including the advantages, challenges, and opportunities, will be discussed, followed by drug release mechanisms and mathematical kinetic models. Subsequently, various modification routes of CNP for improved and enhanced therapeutic efficacy, as well as other restrictions of conventional drug administration for lung cancer treatment, are covered.

A new system composed of chitosan nanoparticles loaded with methotrexate (MTX-CS-NPs) and functionalized with photocatalytic TiO2 nanoparticles (TiO2-NPs) was prepared. This system is expected to initiate polymeric rupture of MTX-CS-NPs and subsequently release MTX, upon illumination with UV light. MTX-CS-NPs were prepared and characterized in terms of particle size, charge, polydispersity and drug release before and after coating with TiO2-NPs. The release of MTX in vitro was studied in dark, light and UV light. Finally, coated and uncoated MTX-CS-NPs were studied in vitro using MCF-7 cell line. The functionalized NPs were larger in size, more polydisperse and carried higher positive charges compared to the unfunctionalized NPs. The entrapment efficacy was high reaching 75% and was not affected by coating with MTX-CS-NPs. Further, less than 5% of methotrexate was released after 80 h from uncoated NPs and the release was not enhanced by UV illumination of the particles. In contrast, the release from functionalized NPs was enhanced, reaching 40% after 80 h, as the particles were stroked with UV light and as the amount of TiO2-NPs used in coating increased. Finally, coating the MTX-CS-NPs with TiO2-NPs significantly enhanced their cytotoxicity on MCF-7 cells. The coated MTX-CS-NPs recorded low cell viabilities compared to the other formulations. In conclusion, the drug release of MTX-CS-NPs could be triggered and controlled remotely by coating with TiO2-NPs, which maybe more effective in cancer treatment.

Chitosan is a natural polysaccharide, mainly derived from the shell of marine organisms. At present, chitosan has been widely used in the field of biomedicine due to its special characteristics of low toxicity, biocompatibility, biodegradation and low immunogenicity. Chitosan nanoparticles can be easily prepared. Chitosan nanoparticles with positive charge can enhance the adhesion of antigens in nasal mucosa and promote its absorption, which is expected to be used for intranasal vaccine delivery. In this study, we prepared chitosan nanoparticles by a gelation method, and modified the chitosan nanoparticles with mannose by hybridization. Bovine serum albumin (BSA) was used as the model antigen for development of an intranasal vaccine. The preparation technology of the chitosan nanoparticle-based intranasal vaccine delivery system was optimized by design of experiment (DoE). The DoE results showed that mannose-modified chitosan nanoparticles (Man-BSA-CS-NPs) had high modification tolerance and the mean particle size and the surface charge with optimized Man-BSA-CS-NPs were 156 nm and +33.5 mV. FTIR and DSC results confirmed the presence of Man in Man-BSA-CS-NPs. The BSA released from Man-BSA-CS-NPs had no irreversible aggregation or degradation. In addition, the analysis of fluorescence spectroscopy of BSA confirmed an appropriate binding constant between CS and BSA in this study, which could improve the stability of BSA. The cell study in vitro demonstrated the low toxicity and biocompatibility of Man-BSA-CS-NPs. Confocal results showed that the Man-modified BSA-FITC-CS-NPs promote the endocytosis and internalization of BSA-FITC in DC2.4 cells. In vivo studies of mice, Man-BSA-CS-NPs intranasally immunized showed a significantly improvement of BSA-specific serum IgG response and the highest level of BSA-specific IgA expression in nasal lavage fluid. Overall, our study provides a promising method to modify BSA-loaded CS-NPs with mannose, which is worthy of further study.