Development of transgenic Paulownia trees expressing antimicrobial thionin genes for enhanced resistance to fungal infections using chitosan nanoparticles.

There is an increasing focus on genetically altering Paulownia trees to enhance their resistance against fungal infections, given their rapid growth and quality wood production. The aim of this research was to establish a technique for incorporating two antimicrobial thionin genes, namely thionin-60 (thio-60) and thionin-63 (thio-63), into Paulownia tomentosa and Paulownia hybrid 9501 through the utilization of chitosan nanoparticles. The outcomes revealed the successful gene transfer into Paulownia trees utilizing chitosan nanoparticles. The effectiveness of thionin proteins against plant pathogens Fusarium and Aspergillus was examined, with a specific focus on Fusarium equiseti due to limited available data. In non-transgenic Paulownia species, the leaf weight inhibition percentage varied from 25 to 36 %, whereas in transgenic species, it ranged from 22 to 7 %. In general, Paulownia species expressing thio-60 displayed increased resistance to F. equiseti, while those expressing thio-63 exhibited heightened resistance to A. niger infection. The thionin proteins displayed a strong affinity for the phospholipid bilayer of the fungal cell membrane, demonstrating their capability to disrupt its structure. The transgenic plants created through this technique showed increased resistance to fungal infections. Thionin-60 demonstrated superior antifungal properties in comparison to thio-63, being more effective at disturbing the fungal cell membrane. These findings indicate that thio-60 holds potential as a novel antifungal agent and presents a promising approach for enhancing the antimicrobial traits of genetically modified Paulownia trees.

Quality by design (QbD) approach-based development of optimized nanocarrier to achieve quality target product profile (QTPP)-targeted lymphatic delivery.

Background.Insulin, commonly used for diabetes treatment, needs better ways to improve its effectiveness and safety due to its challenges with poor permeability and stability. Various system has been developed for oral peptide delivery. The non-targeted system can prevent gastric and enzymatic degradation of peptides but cannot increase the bulk transport of peptides across the membrane. However, the non-selectivity is the limitation of the existing system. Numerous carbohydrate-binding receptors overexpressed on intestinal macrophage cells (M-cells) of gut-associated lymphoid tissue. It is the most desirable site for receptor-mediated endocytosis and lymphatic drug delivery of peptides.Objective. The prime objective of the study was to fabricate mannose ligand conjugated nanoparticles (MNPs) employing a quality-by-design approach to address permeability challenges after oral administration. Herein, the study's secondary objective of this study is to identify the influencing factor for producing quality products. Considering this objective, the Lymphatic uptake of NPs was selected as a quality target product profile (QTPP), and a systematic study was conducted to identify the critical formulation attributes (CFAs) and critical process parameters (CPP) influencing critical quality attributes (CQAs). Mannosylated Chitosan concentrations (MCs) and TPP concentrations were identified as CFAs, and stirring speed was identified as CPP.Methods. MNPs were prepared by the inotropic gelation method and filled into the enteric-coated capsule to protect from acidic environments. The effect of CFAs and CPP on responses like particle size (X) and entrapment (Y) was observed by Box-Behnken design (BBD). ANOVA statistically evaluated the result to confirm a significant level (p< 0.05). The optimal conditions of NPs were obtained by constructing an overlay plot and determining the desirability value. HPLC and zeta-seizer analysis characterized the lyophilized NPs. Cell-line studies were performed to confirm the safety and M-cell targeting of NPs to enhance Insulin oral bioavailability.Results. The morphology of NPs was revealed by SEM. The developed NPs showed a nearly oval shape with the average size, surface potential, and % drug entrapment were 245.52 ± 3.37 nm, 22.12 ± 2.13 mV, and 76.15 ± 1.3%, respectively. MTT assay result exhibited that MNPs safe and Confocal imaging inference that NPs selectively uptake by the M-cell.Conclusion. BBD experimental design enables the effective formulation of optimized NPs. The statistical analysis estimated a clear assessment of the significance of the process and formulation variable. Cell line study confirms that NPs are safe and effectively uptake by the cell.

Synthesis of chitosan polyethylene glycol antibody complex for delivery of Imatinib in lung cancer cell lines.

Lung cancer is known as the most common cancer. Although the Ramucirumab antibody is a second-line treatment for lung cancer, the high interstitial fluid pressure limits the antibody's performance. In this way, Imatinib is a chemotherapeutic drug to reduce the interstitial fluid pressure. Up to now, unfortunately, both Ramucirumab and imatinib have not been reported in one nanosystem for cancer therapy. To fulfill this shortcoming, this paper aims to design a chitosan nanocarrier that loads imatinib and attaches to Ramucirumab for selective bonding to A549. Therefore, this paper aims to develop a polymeric nanosystem for non-small cell lung cancer (NSCLC) treatment. In first, the chitosan polyethylene glycol nanoparticle is synthesized, loaded with imatinib, and then targeted using Ramucirumab. Afterwards, the CS-PEG-Ab-Im by FTIR, TEM, DLS, zeta potential, and TGA techniques are characterized. The size of CS-PEG-Ab-Im was 25-30 nm, its surface charge was 13.1 mV, and the shape of CS-PEG-Ab-Im was nearly spherical and cylindrical. The therapeutic potential of CS-PEG-Ab-Im was assessed using the A549 cell line. According to the obtained results, the cell viability was 48% after 48 h of treatment of A549 cells using the IC50 concentration of CS-PEG-Ab-Im (100 nanomolar). Moreover, the apoptosis and cell cycle arrest percentages were increased by 3 and 6 times, respectively, as compared to free imatinib. Furthermore, the release rate of imatinib from CS-PEG-Ab-Im in an acidic medium was 17% during 1 h, indicating five times the imatinib release in the natural medium. Eventually, the result of flow cytometry indicates the more apoptotic effect of nanosystem to free imatinib and CS-PEG-Ab. Besides, cell arresting result exhibits the CS-PEG-Ab-Im and causes cell arrested at G1 by %8.17. Thus, it can be concluded that CS-PEG-Ab-Im can be an ideal nanosystem in NSCLC treatment.

Effect of final irrigation protocols with chitosan nanoparticle and genipin on dentine against collagenase degradation: An ex-vivo study.

AIM: Endodontic irrigants may affect the mechanical and chemical properties of dentine. This study evaluated the effects of various final irrigation protocols including the use of chitosan nanoparticle (CSnp) and cross-linking with genipin on the (1) mechanical and (2) chemical properties of dentine against enzymatic degradation. METHODOLOGY: CSnp was synthesized and characterized considering physiochemical parameters and stability. The root canals of 90 single-rooted teeth were prepared and irrigated with NaOCl. Dentine discs were obtained and divided into groups according to the following irrigation protocols: Group NaOCl+EDTA, Group NaOCl+CSnp, Group NaOCl+EDTA+CSnp, Group NaOCl+CSnp+Genipin, Group NaOCl+EDTA+CSnp+Genipin and Group distilled water. (1) Mechanical changes were determined by microhardness analysis using Vickers-tester. (2) Chemical changes were determined by evaluating molecular and elemental compositions of dentine using Fourier transform infrared spectroscopy (FTIR) analysis and scanning electron microscope (SEM)/energy dispersive X-ray spectroscopy (EDS) analysis, respectively. All analyses were repeated after the discs were kept in collagenase for 24 h. Data were analysed with repeated measures analysis of variance and Bonferroni correction for microhardness analysis, and Kruskal-Wallis and Wilcoxon tests for FTIR and SEM/EDS analyses (p = .05). RESULTS: (1) Collagenase application did not have a negative effect on microhardness only in Group NaOCl+EDTA+CSnp+Genipin when compared with the post-irrigation values (p > .05). Post-collagenase microhardness of Group NaOCl+EDTA+CSnp and Group NaOCl+CSnp+Genipin was similar to the initial microhardness (p > .05). (2) After collagenase, Amide III/ PO 4 3 - ratio presented no change in Group NaOCl+EDTA+CSnp, Group NaOCl+CSnp+Genipin and Group NaOCl+EDTA+CSnp+Genipin (p > .05), while decreased in other groups (p < .05). Collagenase did not affect CO 3 2 - / PO 4 3 - ratio in the groups (p > .05). There were no changes in the groups in terms of elemental level before and after collagenase application (p > .05). CONCLUSIONS: CSnp and genipin positively affected the microhardness and molecular composition of dentine. This effect was more pronounced when CSnp was used after EDTA.

The effect of the enzymes trypsin and DNase I on the antimicrobial efficiency of root canal irrigation solutions.

The purpose of this study was to evaluate the antibacterial efficacy of using 2.5% NaOCl, 2% chlorhexidine (CHX), Irritrol, and chitosan-coated silver nanoparticles (AgCNPs) alone or in combination with deoxyribonuclease I (DNase I) and trypsin pre-enzyme applications in dentin samples contaminated with Enterococcus faecalis (E. faecalis) by CLSM. 144 dentin blocks with confirmed E. faecalis biofilm formation were divided randomly according to the irrigation protocol (n = 12): NaOCl, CHX, Irritrol, AgCNPs, trypsin before NaOCl, CHX, Irritrol, AgCNPs, and DNase I before NaOCl, CHX, Irritrol, AgCNPs. Dentin blocks were stained with the Live/Dead BacLight Bacterial Viability Kit and viewed with CLSM after irrigation applications. The percentage of dead and viable bacteria was calculated using ImageJ software on CLSM images. At a significance level of p < 0.05, the obtained data were analyzed using one-way Anova and post-hoc Tukey tests. In comparison with NaOCl, CHX had a higher percentage of dead bacteria, both when no pre-enzyme was applied and when DNase I was applied as a pre-enzyme (p < 0.05). There was no difference in the percentage of dead bacteria between the irrigation solutions when trypsin was applied as a pre-enzyme (p > 0.05). AgCNPs showed a higher percentage of dead bacteria when trypsin was applied as a pre-enzyme compared to other irrigation solutions (p < 0.05), while the pre-enzyme application did not affect the percentage of dead bacteria in NaOCl, CHX, and Irritrol (p > 0.05). No irrigation protocol tested was able to eliminate the E. faecalis biofilm. While the application of trypsin as a pre-enzyme improved the antimicrobial effect of AgCNPs, it did not make any difference over other irrigation solutions.

Smear layer removal and antimicrobial efficacy of chitosan as a root canal irrigant: a systematic review of in-vitro studies.

This systematic review was designed to answer the following question: Does chitosan provide better smear layer removal and antimicrobial efficacy than other root canal irrigants? A literature search was done using electronic databases PubMed, Scopus, Web of Science, Cochrane Library, EBSCO host, Grey Literature Report, and Open Grey from inception to June 18, 2024. The reference lists of included articles were also hand-searched. Two reviewers independently assessed the studies' eligibility based on the inclusion and exclusion criteria and performed data extraction. Two reviewers independently evaluated the risk of bias in the selected studies. The search retrieved 2330 studies. After analysis, 36 studies fulfilled the eligibility criteria and were included, with 19 involving smear layer removal, 16 involving antibacterial efficacy, and 1 involving both. The overall risk of bias of the included studies was medium. Chitosan removed the smear layer more effectively than citric acid and acetic acid, similar to MTAD and Qmix, with conflicting results against EDTA. In addition, chitosan demonstrated comparable antibacterial efficacy to chlorhexidine, propolis, and photodynamic therapy but was less effective than sodium hypochlorite. Based on available evidence, it was found that chitosan provided better smear layer removal and antimicrobial efficacy than most root canal irrigants compared in this systematic review. There was substantial heterogeneity in the methodology of included studies. As a result, this review highly recommends further research using standardized methods to assess the effectiveness of chitosan as a root canal irrigant in in-vitro studies to validate its clinical use.

Synthesis and Characterization of Curcumin-Chitosan Loaded Gold Nanoparticles by Oryctes rhinoceros' Chitin for Cosmeceutical Application.

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.

Suppression of dry root rot disease caused by Rhizoctonia bataticola (Taub.) Butler in chickpea plants by application of thiamine loaded chitosan nanoparticles.

The present study evaluated the effect of thiamine loaded chitosan nanoparticle (TChNp) on chickpea plant growth under greenhouse condition. TChNp treated plants showed increased number of leaves, branches, shoot/root length, number of secondary roots and plant dry weight. Enhanced nodulation with larger nodules was observed in TChNp treated chickpea plants. A significant increase in the number of flowers, pods and grain yield was observed in the TChNp treated chickpea plants compared to the chitosan and untreated control. The TChNps showed direct antifungal activity towards Rhizoctonia bataticola as evidenced by in vitro and SEM analyses, which might be due to the solubility and size of the nanoparticle. TChNps treated chickpea plants challenged with R. bataticola showed significant reduction in plant mortality compared to infected untreated control under pot condition. These results indicate the potential of the TChNps in enhancing the yield and suppressing the dry root rot disease in chickpea.

Preparation and Characterization of Salsalate-Loaded Chitosan Nanoparticles: In Vitro Release and Antibacterial and Antibiofilm 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.

Chitosan Nanoparticle-Based System: A New Insight into the Promising Controlled Release System for Lung Cancer Treatment.

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 ß-(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.

Formulation of a pH-sensitive cancer cell-targeted gene delivery system based on folate-chitosan conjugated nanoparticles.

In this study, we investigated the design and construct of a chitosan (CA)-based targeted gene delivery system and evaluated its function. To this end, CA-folic acid/pDNA (CA-FA/pDNA) nanoparticles were prepared in different formulations using the ion gelation method. All the synthesized nanoparticles were characterized using FTIR, TEM, SEM and DLS. Moreover, the effects of molecular weight (MW) of CA, DNA, and CA concentration were inspected on encapsulation efficiency (EE). The results showed that the EE of pDNA was directly proportional with MW of CA and CA concentration but was in an inverse proportion with DNA concentration. In addition, high MW of CA and low MW of CA nanoparticles showed lower and higher pDNA release in all pH ranges, respectively. It is concluded that the N/P ratio increase can cause controlled pDNA release.

Development and Optimization of Chitosan Nanoparticle-Based Intranasal Vaccine Carrier.

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.

Covalently coupling doxorubicin to polymeric nanoparticles as potential inhaler therapy: in vitro studies.

Lung cancer is the most commonly diagnosed type of cancer worldwide, non-small cell lung cancer accounts for most lung cancers. Doxorubicin is a widely used chemotherapy agent in lung cancer. However, the drug has several undesirable side effects. Here, doxorubicin coupled PEGylated mucoadhesive nanoparticles were designed as a doxorubicin delivery system for pH-triggered release in lung cancer therapy through inhaler administration. Firstly, alginate/chitosan nanoparticles were developed at optimum conditions. Then, PEG diacid bound to structures for doxorubicin binding and providing steric hindrance for phagocytosis. Doxorubicin was linked via an acid-labile amide bond to PEGylated nanoparticles and 444.3 ± 9.2 µg doxorubicin was loaded per mg nanoparticle. Doxorubicin coupled PEG diacid linked alginate/chitosan nanoparticles were checked with FTIR. Hydrodynamic diameter and zeta potential of nanoparticles were measured as 205.7 ± 15.0 nm and -25.17 ± 2.67 mV. The morphology of nanoparticles was evaluated as nearly spherical. Drug release studies were performed both in physiological and acidic media. The drug release from nanoparticles reached 23.6% (pH 5.5) and 18% (pH 7.4) within 48 h. The cytotoxicity experiments were done using A549-luc-C8 cells, also statistical analyzes were carried out. The MTT results indicated the designed drug delivery system possessed anti-tumor efficacy for non-small cell lung cancer therapy.

Elicitation of Th1/Th2 related responses in mice by chitosan nanoparticles loaded with Brucella abortus malate dehydrogenase, outer membrane proteins 10 and 19.

Brucella spp. is the causative agent of brucellosis, one of the worldwide diseases. The pathogen infects humans and animals mainly through the digestive or respiratory tract. Therefore, induction of mucosal immunity is required as the first line of defense. In this study, three Brucella abortus recombinant proteins, malate dehydrogenase (rMdh), outer membrane proteins (rOmp) 10 and 19 were loaded in mucoadhesive chitosan nanoparticles (CNs) and induction of mucosal and systemic immunity were investigated after intranasal immunization of BALB/c mice. These antigens were also coimmunized as cocktail (rCocktail) to evaluate multiple antigen specific vaccine candidates. At 6-weeks post-immunization (wpi), antigen specific total IgG was increased in all of the immunized groups, predominantly IgG1. In addition, spleenocyte from rMdh-, rOmp19-, and rCocktail-immunized groups significantly produced IFN-γ and IL-4 suggesting the induction of a mixed Th1-Th2 response. For mucosal immunity, anti-Mdh IgA from nasal washes and fecal excretions, and anti-Omps IgA from sera, nasal washes, genital secretions and fecal excretions were significantly increased in single antigen immunized groups. In the rCocktail-immunized group, anti-Mdh IgA were significantly increased while anti-Omps IgA was not. Collectively, this study indicates that comprise of B. abortus antigen-loaded CNs elicited the antigen-specific IgA with a Th2-polarized immune responses and combination of the highly immunogenic antigens elicited IgG specific to each type of antigen.

Induction of Th2 response through TLR2-mediated MyD88-dependent pathway in human microfold cells stimulated with chitosan nanoparticles loaded with Brucella abortus Mdh.

Drug delivery by the nasal or oral route is considered the preferred route of administration because it can induce systemic mucosal immunity. However, few studies have examined the immunogenicity and transport of antigen at the level of the microfold (M) cell, the epithelial cell that specializes in antigen sampling at mucosal surfaces. In our previous study, Brucella abortus malate dehydrogenase (Mdh) was loaded in chitosan nanoparticles (CNs), and it induced high production of proinflammatory cytokines in THP-1 cells and systemic IgA in BALB/C mice. In the present study, an in vitro M cell model was used in which Caco-2 cells and Raji B cells were co-cultured to investigate the impact of the uptake and immunogenicity of B. abortus Mdh on nanoparticle transport in human M cells. Our results showed that loaded CNs induced enhanced transport of Mdh in the M cell model. ELISAs showed significantly higher production of IL-1ß and IL-6 in the CN-Mdh stimulation group than that seen in the Mdh stimulation group. The observed increase of gene expression of TLR2, MyD88, TRAF6, IRF4 and CD14 implied that MyD88-dependent TLR2 signaling was activated by stimulation with CNs-Mdh. These results suggest that Mdh and CNs may function synergistically to enhance Th2-related responses triggered by the MyD88-dependent TLR2 signaling pathway and could induce an inflammatory response in M cells as an M cell-targeted delivery system. This study will contribute to the development of not only effective antigens for intracellular bacteria, including B. abortus, but also vaccine delivery systems that target M cells.

Structural characterization of chitosan nanoparticle loaded with Piper nigrum essential oil for biological efficacy against the stored grain pest control.

In this study, Piper nigrum essential oil (PNO) has been encapsulated in chitosan nanoparticle (CS NPs) via ionic gelation method with sodium tripolyphosphate (TPP). The successfully loaded Piper nigrum EO was confirmed by UV-Vis spectrophotometry and X-ray diffraction (XRD) techniques. The average particle size of P. nigrum essential oil loaded chitosan nanoparticle (CS/PNO NPs) showed 527.5 nm with spherical shape morphology. Zeta potential values of the particles were found to be negative -5.34 mV. Encapsulation efficiency and loading efficiency was in the range of 35% to 40% and 4.85% to 7.04% respectively. CS/PNO NPs exhibited strong insecticidal activity against Sitophilus oryzae and Tribolium castaneum. In addition, CS/PNO NPs enhanced the fumigant toxicity and altered the neurotransmitter, acetylcholine in both the stored grain pests. Overall results of nanoformulation indicated that these novel design systems could be promoted in integrated pest management schedule for T. castaneum and S. oryzae.

Immunostimulatory effect of dietary chitosan nanoparticles on the performance of Nile tilapia, Oreochromis niloticus (L.).

Feed supplements to fish are generally used to overcome any expected diseases and stressors and to sustain eco-friendly fish farming. One of these feed supplements is chitosan, which stimulated growth and immune properties for many aquatic organisms. It is expected that the nano-sized materials may have stronger immune activation in fish than the ordinary size. Therefore, the current study was conducted to evaluate the effect of dietary chitosan nanoparticles (CNP) on growth performance, antioxidant activity, and innate immunity of Nile tilapia, Oreochromis niloticus (L.). Fish (19.8 ±â€¯0.59 g) were fed on diets enriched with 0.0, 0.25, 0.5, 1.0, and 2.0 g CNP/kg diet for 45 days. Fish performance was significantly improved with increasing CNP levels over the control diet with optimum level of 1.0 g CNP/kg diet. Antioxidant-stimulated activity was observed due to dietary CNP supplementation over the control diet in a dose-dependent manner. However, malondialdehyde level decreased significantly, whereas activities of catalase, superoxide dismutase, lysozyme, and respiratory burst increased significantly due to CNP supplementation in a dose-dependent manner. The current study evoked that dietary CNP showed strong immune modulatory properties and enhanced significantly the performance and health of Nile tilapia with optimum level of 1.0 g CNP/kg diet.

A glutathione responsive nitric oxide release system based on charge-reversal chitosan nanoparticles for enhancing synergistic effect against multidrug resistance tumor.

The combination therapy of nitric oxide (NO) and anticancer drug was developed for reversing multidrug resistance (MDR). In order to avoid NO release during the blood circulation, and realize pinpointed release in the tumor cells, we designed a tumor-specific NO-release system based on 10-hydroxycamptothecin (HCPT)-loaded charge-reversal chitosan nanoparticles and covalently linked phenylsulfonyl furoxan (glutathione (GSH)-responsive NO donor) on the surface. The results showed that only 6.0% of NO was eventually released under physiology condition (pH 7.4 and 2 µM GSH) within 8 h. In contrast, 93.0% of NO was released within 4 h in the presence of 10 mM GSH. Western blot result displayed the P-glycoprotein expression was significantly decreased by 50.1%. Hence, this system performed remarkable cytotoxicity in vitro and the highest tumor inhibition rate (79.7%) comparing with free HCPT group (20.7%) in vivo. Such GSH-responsive NO-release system is a promising candidate with prominent therapeutic effect against MDR tumor.

Implementation of two different experimental designs for screening and optimization of process parameters for metformin-loaded carboxymethyl chitosan formulation.

Metformin (MET) was effectively encapsulated into O-carboxymethyl chitosan (O-CMC) polymeric formulation using an experimental design method. Six factors Plackett-Burman (PB) design was utilized to find the significant process parameters. Linear equations used to study the effect of each process parameters on particle size (PS), encapsulation efficiency (EE), and zeta potential (ZP) and the most influential three factors decided for further optimization. Optimization was carried out by implementing three-factor three-level Box-Behnken (BB) design. Mathematical models were generated by regression analysis for responses of PS, EE, and ZP. Two-step experimental design took into account for the preparation of optimized formulation with maximum %EE (72.78 ± 9.7%) and minimum PS (225.67 ± 5.53 nm) at optimum process conditions with a ZP of -5.22 mV for the nano-polymeric formulation in an economical matter by reduction chemical use and formulation time. Furthermore, the biological activity of the final formulation was determined by in vitro cytotoxicity study compared to free MET. The cytotoxicity result reveals that both pure drug and nano-formulation biocompatible with MCF10A non-tumorigenic cell line and lethal for the MCF7 cell line. These in vitro results were the first helpful step to further investigate O-CMC loaded MET nanoparticles in diagnostic and therapeutic applications of breast cancer.

Effect of chitosan nanoparticle, QMix, and EDTA on TotalFill BC sealers' dentinal tubule penetration: a confocal laser scanning microscopy study.

The aim of the present study was to compare the effect of chitosan nanoparticle, QMix, and 17% EDTA on the penetrability of a calcium silicate-based sealer into dentinal tubules using a confocal laser scanning microscope (CLSM). Sixty mandibular premolar teeth were selected and randomly divided into three groups (n = 20) before root canal preparation according to the solution used in the final rinse protocol: chitosan, QMix, and EDTA groups. Twenty teeth of each group were filled with a TotalFill BC sealers' single gutta-percha cone and with 0.1% rhodamine B. The specimens were horizontally sectioned at 3 and 5 mm from the apex, and the slices were analyzed in CLSM (4×). Total percentage and maximum depth of sealer penetration were measured using confocal laser scanning microscopy with using Image J analysis software. Dentinal tubule's penetration depth, percentage, and area were measured using imaging software. Kruskal-Wallis test was used for statistical analysis. The level of significance was set at 5%. Results of Kruskal-Wallis analysis showed that there was a significant difference in the percentage and depth of sealer penetration among all groups at 3 and 5 mm level sections (P < 0.05). Within the groups, the minimum sealer penetration depth was recorded for chitosan nanoparticle group. Greater depth of sealer penetration was recorded at 5 mm as compared to 3 mm in all the groups. Within the limitation of the present study, it can be concluded that QMix and EDTA promoted sealer penetration superior to that achieved by chitosan nanoparticle.