Objective:To improve the quality of post-thawing Boer buck semen for artificial insemination by adding green tea extract chitosan nanoparticles to skimmed egg yolk diluent,and the proper thawing temperature.Methods:Th...Objective:To improve the quality of post-thawing Boer buck semen for artificial insemination by adding green tea extract chitosan nanoparticles to skimmed egg yolk diluent,and the proper thawing temperature.Methods:The ejaculate of Boer buck was added to skimmed egg yolk diluent without(the control group)and with adding 1μg of chitosan nanoparticles of green tea extract per mL of diluent(the treatment group).Then,the diluted semen was filled in French mini straws containing 60×106 live sperm per straw,frozen in a standard protocol,and stored as frozen semen at−196℃for a week.Six replicates from each group were diluted for 30 s at 37℃or 39℃sterile water to evaluate the semen quality.Results:Post-thawing(at 37℃or 39℃)of live sperm,progressive motility,and plasma membrane integrity were lower compared to those of the pre-freezing stage(P<0.05).Thawing at 37℃resulted in no significant difference in live sperm,progressive motility,and plasma membrane between the control group and the treatment group(P>0.05).The live sperm,progressive motility,and plasma membrane of the treatment group in the pre-freezing stage,and post-thawed at 39℃were higher compared to those of the control group(P<0.05).There was no significant difference in malondialdehyde(MDA)concentration,DNA fragmentation,and catalase concentration of thawing at 37℃compared to those of 39℃in the same group.The MDA concentration and DNA fragmentation in thawing at 37℃and 39℃of the treatment group were significantly lower than those of the control group(P<0.05).However,the catalase concentration in thawing at 37℃and 39℃of the treatment group was not significantly different than the control group(P>0.05).Conclusions:Higher quality post-thawing Boer buck semen is achieved by adding 1μg/mL of chitosan nanoparticles of green tea extract to the skimmed egg yolk diluent and thawing at 39℃.展开更多
Silver nanoparticles(Ag NPs)are an effective antibacterial agent,but their application in food packaging is limited due to their easy agglomeration and oxidation.In this study,antibacterial microcapsules were fabricat...Silver nanoparticles(Ag NPs)are an effective antibacterial agent,but their application in food packaging is limited due to their easy agglomeration and oxidation.In this study,antibacterial microcapsules were fabricated using Ginkgo biloba essential oil(GBEO)as core material and chitosan and type B gelatin biopolymer as capsule mate-rials.These antibacterial microcapsules were then modified with green-synthesized Ag NPs,blended into the bio-polymer polylactic acid(PLA),and finally formed as films.Physicochemical properties and antibacterial activity against Escherichia coli(E.coli)and Staphylococcus aureus(S.aureus)were evaluated.Results showed that the prepared antibacterial PLA films exhibited excellent antibacterial activity against foodborne pathogens.Its TVC exceeded the limit value of 7 log CFU/g at 7 days compared with the 5 days of pure PLA films.Therefore,these films can extend the shelf life of grass carp fillets by 2–3 days under refrigeration.展开更多
Cetyl-chitosan, prepared by reacting chitosan with chlorocetane under alkaline condition, is soluble and spontaneously forms nanoparticles about 100 nm in diameter. Infrared spectra (IR) revealed that there was a subs...Cetyl-chitosan, prepared by reacting chitosan with chlorocetane under alkaline condition, is soluble and spontaneously forms nanoparticles about 100 nm in diameter. Infrared spectra (IR) revealed that there was a substitution reaction mainly on the amine groups of chitosan (CS). By using paracetamol (PCTM) as a model drug, the balanced release concentration of PCTM in phosphate buffer solution (pH=7.4) can be decreased with the increase of degree of substitution alkyl and can be reduced effectively even under a lower PCTM loading.展开更多
Galactosylated chitosan (GC) is synthesized and used to prepare IL-1Ra loaded GC nanoparticles by an electrospraying technique. Polyethylene oxide (PEO) is mixed with GC to enhance the electrospraying ability. The...Galactosylated chitosan (GC) is synthesized and used to prepare IL-1Ra loaded GC nanoparticles by an electrospraying technique. Polyethylene oxide (PEO) is mixed with GC to enhance the electrospraying ability. The effect of the spraying solution properties on particle formation is investigated. The IL-1Ra loaded nanoparticles with an average diameter of 530 nm and a regularly spherical shape are observed by the scanning electron microscopy (SEM). The amount of the IL-1Ra is measured by the enzyme-linked immunosorbent assay (ELISA) kit. The loading capacity of the nanoparticle is (1.52± 0.04)% (n = 3) and the encapsulation efficiency reaches (90. 36 ± 3.46) % (n = 3). For the evaluation of GC nanoparticles' hepatocytes targeting efficacy, hepatocytes and mesenchymal stem cells (MSCs) are incubated with FITC-labeled GC nanoparticles for 24 h as the experimental and control groups. Results of the fluorescence microscope show that the fluorescence signals observed in hepatocytes are significantly higher than in the MSCs, indicating that the developed GC nanoparticles have an obvious liver targeting property.展开更多
Epilepsy is considered as a common and diverse set of chronic neurological disorders and its symptoms can be controlled by antiepileptic drugs(AEDs). The presence of p-glycoprotein and multi-drug resistance transporte...Epilepsy is considered as a common and diverse set of chronic neurological disorders and its symptoms can be controlled by antiepileptic drugs(AEDs). The presence of p-glycoprotein and multi-drug resistance transporters in the blood-brain barrier could prevent the entry of AEDs into the brain, causing drug resistant epilepsy. To overcome this problem, we propose using carboxymethyl chitosan nanoparticles as a carrier to deliver carbamazepine(CBZ) intranasally with the purpose to bypass the blood-brain barrier thus to enhance the brain drug concentration and the treatment efficacy. Results so far indicate that the developed CBZNPs have small particle size(218.76 ± 2.41 nm) with high drug loading(around 35%) and high entrapment efficiency(around 80%). The in vitro release profiles of CBZ from the NPs are in accordance with the Korsmeyer-peppas model. The in vivo results show that both encapsulation of CBZ in nanoparticles and the nasal route determined the enhancement of the drug bioavailability and brain targeting characteristics.展开更多
The magnetic chitosan nanoparticles were prepared by reversed-phase suspension method using Span-80 as an emulsifier, glutaraldehyde as cross-linking reagent. And the nanoparticles were characterized by TEM, FT-IR and...The magnetic chitosan nanoparticles were prepared by reversed-phase suspension method using Span-80 as an emulsifier, glutaraldehyde as cross-linking reagent. And the nanoparticles were characterized by TEM, FT-IR and hysteresis loop. The results show that the nanoparticles are spherical and almost superparamagnetic. The laccase was immobilized on nanoparticles by adsorption and subsequently by cross-linking with glutaraldehyde. The immobilization conditions and charac-terizations of the immobilized laccase were investigated. The optimal immobilization conditions were as follows: 10 mL of phosphate buffer (0.1 M, pH 7.0) containing 50 mg of magnetic chitosan nanoparticles, 1.0 mg·mL-1 of laccase and 1% (v/v) glutaraldehyde, immobilization temperature of 4 ℃ and immobilization time of 4 h. The immobilized laccase exhibited an appreciable catalytic capability (480 units·g-1 support) and had good storage stability and operation stability. The Km of immobilized and free laccase for ABTS were 140.6 and 31.1 μM in phosphate buffer (0.1 M, pH 3.0) at 37 ℃, respectively. The immobilized laccase is a good candidate for the research and development of biosensors based on laccase catalysis.展开更多
pH-responsive-chitosan nanoparticles for the control release of protein drug were prepared by combining two-step crosslinking method, in which chitosan was subsequently crosslinked by sodium tripolyphosphate (TPP) a...pH-responsive-chitosan nanoparticles for the control release of protein drug were prepared by combining two-step crosslinking method, in which chitosan was subsequently crosslinked by sodium tripolyphosphate (TPP) and glycidoxypropyltrimethoxysilane (GPTMS). Compared with TPP crosslinked chitosan particles, the two-step crosslinked nanoparticles were not only pH-responsive but also more stable in wide pH range. Fluorescein isothiocyanate (FITC) labeled anti-human-IgG antibody was used as a model protein drug for evaluating the control release profile of the nano-carrier. The amount of released antibody increased from 5.6% to 50% when the pH of solution shifted from 7.4 to 6.0. The results suggest the possible application of the nanoparticles as pH- responsive drug delivery materials.展开更多
AIM: To prepare chitosan-polyaspartic acid-5-fluorouracil (CTS-Pasp-5Fu) nanoparticles and investigate its anti-carcinoma effect and toxicity. METHODS: CTS-Pasp-5Fu nanoparticles were synthesized by ionic gelatificati...AIM: To prepare chitosan-polyaspartic acid-5-fluorouracil (CTS-Pasp-5Fu) nanoparticles and investigate its anti-carcinoma effect and toxicity. METHODS: CTS-Pasp-5Fu nanoparticles were synthesized by ionic gelatification. Male BABL/c nude mice were injected with SGC-7901 gastric carcinoma cell line mass to establish a human gastric carcinoma model. They were randomly allocated into 4 groups: CTS-Pasp-5Fu (containing 5-Fu 1.25 mg/kg), 5-Fu (1.25 mg/kg), CTS-Pasp and normal saline groups. Tumor weight was measured and assay of colony forming unit-granulocyte and macrophage (CFU-GM) was performed. The structural change of cells and tissues was observed and the Bax and Bcl-2 genes were detected. RESULTS: Compared with normal saline, the inhibition rates of tumor growth for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.58%, 58.69% and 70.82%, respectively. The tumor inhibition rates for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.09%, 65.3% and 72.79%, respectively. There was a significant decrease in the number of CFU-GMformation and increase of total bilirubin, and alanine aminotransferase in the 5-Fu group, but no change in those of the other three groups. There was no change in white blood cell count and creatinine among the four groups. Pathological section of liver and nephridial tissues showed that the damage in the 5-Fu group was more severe than that in the CTS-Pasp-5Fu group. 5-Fu and CTS-Pasp-5Fu groups could both down-regulate the Bcl-2 expression and up-regulate the Bax expression to different extent, and the accommodate effect of CTS-Pasp-5Fu was more obvious than 5-Fu. CONCLUSION: The tumor inhibition rate of CTS-Pasp-5Fu nanoparticles is much higher than that of 5-Fu alone.展开更多
The sustained-release properties of the biodegradable nano-drug delivery systems were used to improve the residence time of the chemotherapeutic agent in the body. These drug delivery systems were widely used to deliv...The sustained-release properties of the biodegradable nano-drug delivery systems were used to improve the residence time of the chemotherapeutic agent in the body. These drug delivery systems were widely used to deliver chemotherapeutic drugs. The 5-fluorouracil loaded chitosan nanoparticles prepared in this paper have the above advantage. Here, we found that when the mass ratio of 5-fluorouracil and chitosan was 1:1, the maximum drug loading of nanoparticles was 20.13 ± 0.007%, the encapsulation efficiency was 44.28 ± 1.69%, the particle size was 283.9 ± 5.25 nm and the zeta potential was 45.3 ± 3.23 mV. The prepared nanoparticles had both burst-release and sustained-release phases in vitro release studies.In addition, the inhibitory effect of the prepared nanoparticles on gastric cancer SGC-7901 cells was similar to that of 5-fluorouracil injection, and the blank vector had no obvious inhibitory effect on SGC-7901 cells. In the pharmacokinetic study of rats in vivo, we found that AUC(0-t), MRT(0-t) and t1/2 z of nanoparticles were significantly increased in vivo compared with 5-fluorouracil solution, indicating that the prepared nanoparticles can play a role in sustained-release.展开更多
Chitosan was modified by conjugating coupling with linolenic acid through the 1-ethyl-3-(3-dimethylami- nopropyyl) carbodiimide (EDC)-mediated reaction. The degree of substitution 1.8% ( i.e. 1.8 linolenic acid g...Chitosan was modified by conjugating coupling with linolenic acid through the 1-ethyl-3-(3-dimethylami- nopropyyl) carbodiimide (EDC)-mediated reaction. The degree of substitution 1.8% ( i.e. 1.8 linolenic acid group per 100 anhydroglucose units) was measured by ^1H NMR. The critical aggregation concentration (CAC) of the self-aggregate of hydrophobically modified chitosan was determined by measuring the fluorescence intensity of the pyrene as a fluorescent probe. The CAC value in phosphate-buffered saline (PBS) solution (pH 7.4) was 5 × 10^-2 mg mL^-1. The average particle size of selfaggregates of hydrophobically modified chitosan in PBS solution (pH7.4) was 210.8 nm with a unimodal size distribution ranging from 100 to 500 nm. Transmission electron microscopy (TEM) study showed that the formation of near spherical shape nanoparticles has enough structural integrity. The loading ability of hydrophibically modified chitosan (LA-chitosan) was investigated by using bovine serum albumin (BSA) as the model. The loading capacity of self-aggregated nanoparticles increases ( 19.85 % ± 0.04 % to 37.57 % ± 0.25 % ) with the concentration of BSA (0.1-0.5 mg mL^-1 ).展开更多
The aim of this paper is to investigate and optimize the preparation of scutellarin(SCU)-loaded HP-β-CD/chitosan(CS) nanoparticles(CD/CS-SCU-NPs). CD/CS-SCU-NPs were prepared by ionic cross-linking method and the pro...The aim of this paper is to investigate and optimize the preparation of scutellarin(SCU)-loaded HP-β-CD/chitosan(CS) nanoparticles(CD/CS-SCU-NPs). CD/CS-SCU-NPs were prepared by ionic cross-linking method and the process and formulation variables were optimized using response surface methodology(RSM) with a three-level, three factor Box–Behnken design(BBD).The independent variables were the added amounts of CS, sodium tripolyphosphate(TPP)and Pluronic F-68 during the preparation. Dependent variables(responses) were particle size and entrapment efficiency. Mathematical equations and respond surface plots were used to correlate independent and dependent variables.The preparation process and formulation variables were optimized to achieve minimum particle size and maximum entrapment efficiency by calculating the overall desirability value(OD). The optimized NP formulation was characterized for particle size, PDI, zeta potential, entrapment efficiency and in vitro drug release.According to the results, an optimized CD/CS-SCU-NP formulation was prepared. Results for particle size, PDI, zeta potential and entrapment efficiency were found to be around 200 nm,0.5, 25 mV, and 70% respectively. For in vitro study, the release of SCU from the NPs exhibited a biphasic release and was in accordance with Higuchi equation. The optimized preparation was simple with the probability for industrialization. The combination use of RSM, BBD and overall desirability values could provide a promising application for incorporating CD into CS nanoparticles as drug delivery carrier and help develop lab-scale procedures.展开更多
AIM: To investigate the effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 in vitro and the possible mechanisms involved. METHODS: Chitosan nanoparticles were characterize...AIM: To investigate the effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 in vitro and the possible mechanisms involved. METHODS: Chitosan nanoparticles were characterized by particle size, zeta potential, and morphology. After treatment with various concentrations of chitosan nanoparticles (25, 50, 75, 100 μg/mL) at various time intervals, cell proliferation, ultrastructural changes, DNA fragmentation, mitochondrial membrane potential (MMP), cell cycle phase distribution and apoptotic peaks of MGC803 cells were analyzed by MTT assay, electron microscopy, DNA agarose gel electrophoresis, and flow cytometry. RESULTS: Chitosan nanoparticles exhibited a small particle size as 65 nm and a high surface charge as 52 mV. Chitosan nanoparticles markedly inhibited cell proliferation of MGC803 cells with an ICso value of 5.3 μg/mL 48 h after treatment. After treatment with chitosan nanoparticles, the typical necrotic cell morphology was observed by electron microscopy, a typical DNA degradation associated with necrosis was determined by DNA agarose electrophoresis. Flow cytometry showed the loss of MMP and occurrence of apoptosis in chitosan nanoparticles-treated cells. CONCLUSION: Chitosan nanoparticles effectively inhibit the proliferation of human gastric carcinoma cell line MGC803 in vitro through multiple mechanisms, and may be a beneficial agent against human carcinoma.展开更多
β-cyclodextrin (CD) grafted N-maleoyl chitosan (CD-g-NMCS) with two different degrees of substitution (DS) of N-maleoyl (DS = 21.2% and 30.5%) were synthesized from maleic anhydride and chitosan bearing pendant cyclo...β-cyclodextrin (CD) grafted N-maleoyl chitosan (CD-g-NMCS) with two different degrees of substitution (DS) of N-maleoyl (DS = 21.2% and 30.5%) were synthesized from maleic anhydride and chitosan bearing pendant cyclodextrin (CD-g-CS). CD-g-NMCS based nanoparticles were prepared via an ionic gelation method together with chitosan and CD-g-CS nanoparticles.The size and zeta potential of prepared CD-g-NMCS nanoparticles were 179.2~274.0 nm and 36.2~42.4 m V, respectively. In vitro stability test indicated that CD-g-NMCS nanoparticles were more stable in phosphate-buffered saline compared with chitosan nanoparticles. Moreover, a poorly water-soluble drug, ketoprofen (KTP), was selected as a model drug to study the obtained nanoparticle’s potentials as drug delivery carriers. The drug loading efficiency of CD-g-NMCS20 nanoparticles were 14.8% for KTP. MTT assay showed that KTP loaded CD-g-NMCS nanoparticles were safe drug carriers. Notably, in vitro drug release studies showed that KTP was released in a sustained-release manner for the nanoparticles. The pharmacokinetic of drug loaded CD-g-NMCS20 nanoparticles were evaluated in rats after intravenous administration. The results of studies revealed that, compared with free KTP, KTP loaded CD-g-NMCS20 nanoparticles exhibited a significant increase in AUC0→24h and mean residence time by 6.6-fold and 2.9-fold, respectively. Therefore, CD-g-NMCS nanoparticles could be used as a novel promising nanoparticle-based drug delivery system for sustained release of poorly water-soluble drugs. The carboxylic acid groups of the CD-g-NMCS molecule provide convenient sites for further structural modifications including introduction of tissue-or disease-specific targeting groups.展开更多
Objective To develop a safe and novel immunoadjuvant to enhance the immunity and resistance of animals against E. coli infection. Methods An 88-base immunostimulatory oligodeoxynuleotide containing eleven CpG motifs ...Objective To develop a safe and novel immunoadjuvant to enhance the immunity and resistance of animals against E. coli infection. Methods An 88-base immunostimulatory oligodeoxynuleotide containing eleven CpG motifs (CpG ODN) was synthesized and amplified by PCR. The chitosan nanoparticle (CNP) was prepared by ion linking method to entrap the CpG ODN that significantly promotes the proliferation of lymphocytes of pig in vitro. Then the CpG- CNP was inoculated into 21-day old Kunming mice, which were orally challenged with virulent K88/K99 E. Coil 35 days after inoculation. Blood was collected from the tail vein of mice on days 0, 7, 14, 21, 28, 35, 42, and 49 after inoculation to detect the changes and content of immunoglobulins, cytokines and immune cells by ELISA, such as IgG, IgA, IgM, IL-2, IL-4, and IL-6. Results The CpG provoked remarkable proliferation of lymphocytes of pig in vitro in comparison with that of control group (P〈0.05). The inoculation with CpG-CNP significantly raised the content of IgG, IgM, and IgA in the sera of immunized mice (P〈0.05). The levels of IL-2, 1L-4, and IL-6 in the mice significantly increased in comparison with those in controls (P〈0.05), so was the number of white blood cells and lymphocytes in immunized mice. The humoral and cellular immunities were significantly enhanced in immunized mice, which resisted the infection of E coli and survived, while the control mice manifested evident symptoms and lesions of infection. Conclusions CpG-CNP can significantly promote cellular and humoral immunity and resistance of mice against E. coli infection, and can be utilized as an effective adjuvant to improve the immunoprotection and resistance of porcine against infectious disease.展开更多
Gene-based therapeutics has emerged as a promising approach for human cancer therapy. Among a variety of non-viral vectors, polymer vectors are particularly attractive due to their safety and multivalent groups on the...Gene-based therapeutics has emerged as a promising approach for human cancer therapy. Among a variety of non-viral vectors, polymer vectors are particularly attractive due to their safety and multivalent groups on their surface. This study focuses on guanidinylated O-carboxymethyl chitosan(GOCMCS) along with poly-β-amino ester(PBAE) for si RNA delivery. Binding efficiency of PBAE/si RNA/GOCMCS nanoparticles were characterized by gel electrophoresis. The si RNA-loaded nanoparticles were found to be stable in the presence of RNase A, serum and BALF respectively. Fine particle fraction(FPF) which was determined by a two-stage impinger(TSI) was 57.8% ± 2.6%. The particle size and zeta potential of the nanoparticles were 153.8 ± 12.54 nm and + 12.2 ± 4.94 m V. In vitro cell transfection studies were carried out with A549 cells. The cellular uptake was significantly increased. When the cells were incubated with si Survivin-loaded nanoparticles, it could induce 26.83% ± 0.59% apoptosis of A549 cells and the gene silencing level of survivin expression in A549 cells were 30.93% ± 2.27%. The results suggested that PBAE/GOCMCS nanoparticle was a very promising gene delivery carrier.展开更多
Chitosan is a natural polysaccharide prepared by the N-deacetylation of chitin. In this paper we have reviewed the methods of preparation of chitosan-based nanoparticles and their pharmaceutical applications. There ar...Chitosan is a natural polysaccharide prepared by the N-deacetylation of chitin. In this paper we have reviewed the methods of preparation of chitosan-based nanoparticles and their pharmaceutical applications. There are five methods of their prepa-rations:emulsion cross-linking,emulsion-droplet coalescence,ionic gelation,reverse micellar method and chemically modified chi-tosan method. Chitosan nanoparticles are used as carriers for low molecular weight drug,vaccines and DNA. Releasing characteris-tics,biodistribution and applications are also summarized.展开更多
Chitosan nanoparticles are exhalation prone and agglomerative to pulmonary inhalation.Blending nanoparticles with lactose microparticles(~5 μm) could mutually reduce their agglomeration through surface adsorption phe...Chitosan nanoparticles are exhalation prone and agglomerative to pulmonary inhalation.Blending nanoparticles with lactose microparticles(~5 μm) could mutually reduce their agglomeration through surface adsorption phenomenon. The chitosan nanoparticles of varying size, size distribution, zeta potential, crystallinity, shape and surface roughness were prepared by spray drying technique as a function of chitosan, surfactant and processing conditions. Lactose-polyethylene glycol 3000(PEG3000) microparticles were similarly prepared. The chitosan nanoparticles, physically blended with fine lactose-PEG3000 microparticles, exhibited a comparable inhalation performance with the commercial dry powder inhaler products(fine particle fraction between 20% and 30%). Cascade impactor analysis indicated that the aerosolization and inhalation performance of chitosan nanoparticles was promoted by their higher zeta potential and circularity, and larger size attributes of which led to reduced inter-nanoparticulate aggregation and favored nanoparticles interacting with lactose-PEG3000 micropaticles that aided their delivery into deep and peripheral lungs.展开更多
Chitosan(CS)nanofibers containing silver nanoparticles(AgNPs)were prepared by in-situ reducing method.A water soluble carboxymethyl chitosan(CMCT)was applied for the preparation of AgNPs.The impact factor such as the ...Chitosan(CS)nanofibers containing silver nanoparticles(AgNPs)were prepared by in-situ reducing method.A water soluble carboxymethyl chitosan(CMCT)was applied for the preparation of AgNPs.The impact factor such as the concentration of CMCT,silver nitrate(AgNO_3)content,temperature and the heating time during the preparation of AgNPs were studied.The result showed that the proper value of the concentration of CMCT,AgNO_3content,temperature and the heating time were set as0.1%,20μL AgNO_3(1.7 mol/L),90°and 3 h,separately and the maximum concentration of AgNPs could be acquired.To solve the spinnability of chitosan nanofiber,a super high molecular weight polyethylene oxide(PEO)was introduced to the system,and a new mixed solvent system was prepared by adding acetic acid,dimethyl sulfoxide(DMSO)and several drops of Triton X-100TMto distilled water.CS/PEO(80/20)with the concentration of 3%was dissolved in the mixed solvent to prepare electrospinning solution for CS/PEO(80/20)nanofiber fabrication.The CS containing AgNPs electrospun solution could be prepared by replacing the distilled water to silver nanoparticle solution during the preparation of mixed solvent.Ultraviolet visible(UV-Vis)spectra and transmission electron microscope(TEM)results showed that silver nanoparticles were prepared successfully.CS membranes with and without AgNPs were acquired via a traditional electrospinning equipment.These two nanofiber membranes were characterized by scanning electron microscope(SEM)images and mechanical testing.It could be noticed from the SEM images that there was a good morphology and random distribution for the nanofibers with an average fiber diameter of 180 nm.The mechanical property results showed that the addition of AgNPs decreased the mechanical strength significantly but the mechanical strength could still support wound dressing application.展开更多
AIM:To study the expression of human insulin gene in gastrointestinal tracts of diabetic rats. METHODS: pCMV.Ins, an expression plasmid of the human insulin gene, wrapped with chitosan nanoparticles, was transfected t...AIM:To study the expression of human insulin gene in gastrointestinal tracts of diabetic rats. METHODS: pCMV.Ins, an expression plasmid of the human insulin gene, wrapped with chitosan nanoparticles, was transfected to the diabetic rats through lavage and coloclysis, respectively. Fasting blood glucose and plasma insulin levels were measured for 7 d. Reverse transcription polymerase chain reaction (RT-PCR) analysis and Western blot analysis were performed to confirm the expression of human insulin gene. RESULTS: Compared with the control group, the fasting blood glucose levels in the lavage and coloclysis groups were decreased significantly in 4 d (5.63 ± 0.48 mmol/L and 5.07 ± 0.37 mmol/L vs 22.12 ± 1.31 mmol/L, respectively, P < 0.01), while the plasma insulin levels were much higher (32.26 ± 1.81 μIU/mL and 32.79 ± 1.84 μIU/mL vs 14.23 ± 1.38 μIU/mL, respectively, P < 0.01). The human insulin gene mRNA and human insulin were only detected in the lavage and coloclysis groups. CONCLUSION: Human insulin gene wrapped with chitosan nanoparticles can be successfully transfected to rats through gastrointestinal tract, indicating that chitosan is a promising non-viral vector.展开更多
Folic acid conjugated chitosan was prepared by cross-linking reaction with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC), and then used as a template to prepare folic acid-chitosan(FA-CS) conjugate...Folic acid conjugated chitosan was prepared by cross-linking reaction with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC), and then used as a template to prepare folic acid-chitosan(FA-CS) conjugated nanoparticles and load mitoxantrone nanoparticles(FA-CSNP/MTX). Drug dissolution testing, CCK-8 method, and confocal microscopy were used to detect their controlled-release capability in different situations and the specific uptake by HONE1 cells. The experimental results show that the nanoparticles have uniform size distribution of 48-58 nm. The highest encapsulation rate of the particles on mitoxantrone hydrochloride(MTX) is(77.5±1.9)%, and the drug loading efficiency is(18.4±0.4)%. The sustained release effect, cell growth inhibition activity and targeting effect of the FA-CS/MTX nanoparticles are good in artificial gastric fluid and intestinal fluid. It is demonstrated that the FA-CSNP system is a potentially useful system for the targeted delivery of anticancer drug MTX.展开更多
基金funded by Universitas Airlangga,Indonesia,contract number:1405/UN3.1.6/PT/2022.
文摘Objective:To improve the quality of post-thawing Boer buck semen for artificial insemination by adding green tea extract chitosan nanoparticles to skimmed egg yolk diluent,and the proper thawing temperature.Methods:The ejaculate of Boer buck was added to skimmed egg yolk diluent without(the control group)and with adding 1μg of chitosan nanoparticles of green tea extract per mL of diluent(the treatment group).Then,the diluted semen was filled in French mini straws containing 60×106 live sperm per straw,frozen in a standard protocol,and stored as frozen semen at−196℃for a week.Six replicates from each group were diluted for 30 s at 37℃or 39℃sterile water to evaluate the semen quality.Results:Post-thawing(at 37℃or 39℃)of live sperm,progressive motility,and plasma membrane integrity were lower compared to those of the pre-freezing stage(P<0.05).Thawing at 37℃resulted in no significant difference in live sperm,progressive motility,and plasma membrane between the control group and the treatment group(P>0.05).The live sperm,progressive motility,and plasma membrane of the treatment group in the pre-freezing stage,and post-thawed at 39℃were higher compared to those of the control group(P<0.05).There was no significant difference in malondialdehyde(MDA)concentration,DNA fragmentation,and catalase concentration of thawing at 37℃compared to those of 39℃in the same group.The MDA concentration and DNA fragmentation in thawing at 37℃and 39℃of the treatment group were significantly lower than those of the control group(P<0.05).However,the catalase concentration in thawing at 37℃and 39℃of the treatment group was not significantly different than the control group(P>0.05).Conclusions:Higher quality post-thawing Boer buck semen is achieved by adding 1μg/mL of chitosan nanoparticles of green tea extract to the skimmed egg yolk diluent and thawing at 39℃.
基金supported by the National Key R&D Program of China(Grant No.2020YFD0900905).
文摘Silver nanoparticles(Ag NPs)are an effective antibacterial agent,but their application in food packaging is limited due to their easy agglomeration and oxidation.In this study,antibacterial microcapsules were fabricated using Ginkgo biloba essential oil(GBEO)as core material and chitosan and type B gelatin biopolymer as capsule mate-rials.These antibacterial microcapsules were then modified with green-synthesized Ag NPs,blended into the bio-polymer polylactic acid(PLA),and finally formed as films.Physicochemical properties and antibacterial activity against Escherichia coli(E.coli)and Staphylococcus aureus(S.aureus)were evaluated.Results showed that the prepared antibacterial PLA films exhibited excellent antibacterial activity against foodborne pathogens.Its TVC exceeded the limit value of 7 log CFU/g at 7 days compared with the 5 days of pure PLA films.Therefore,these films can extend the shelf life of grass carp fillets by 2–3 days under refrigeration.
文摘Cetyl-chitosan, prepared by reacting chitosan with chlorocetane under alkaline condition, is soluble and spontaneously forms nanoparticles about 100 nm in diameter. Infrared spectra (IR) revealed that there was a substitution reaction mainly on the amine groups of chitosan (CS). By using paracetamol (PCTM) as a model drug, the balanced release concentration of PCTM in phosphate buffer solution (pH=7.4) can be decreased with the increase of degree of substitution alkyl and can be reduced effectively even under a lower PCTM loading.
基金The National Natural Science Foundation of China(No.30901431)the Natural Science Foundation of Jiangsu Province(No.BK2010242)
文摘Galactosylated chitosan (GC) is synthesized and used to prepare IL-1Ra loaded GC nanoparticles by an electrospraying technique. Polyethylene oxide (PEO) is mixed with GC to enhance the electrospraying ability. The effect of the spraying solution properties on particle formation is investigated. The IL-1Ra loaded nanoparticles with an average diameter of 530 nm and a regularly spherical shape are observed by the scanning electron microscopy (SEM). The amount of the IL-1Ra is measured by the enzyme-linked immunosorbent assay (ELISA) kit. The loading capacity of the nanoparticle is (1.52± 0.04)% (n = 3) and the encapsulation efficiency reaches (90. 36 ± 3.46) % (n = 3). For the evaluation of GC nanoparticles' hepatocytes targeting efficacy, hepatocytes and mesenchymal stem cells (MSCs) are incubated with FITC-labeled GC nanoparticles for 24 h as the experimental and control groups. Results of the fluorescence microscope show that the fluorescence signals observed in hepatocytes are significantly higher than in the MSCs, indicating that the developed GC nanoparticles have an obvious liver targeting property.
基金the Academic Research Fund,Faculty of Science,National University of Singapore,R148-000-180-112
文摘Epilepsy is considered as a common and diverse set of chronic neurological disorders and its symptoms can be controlled by antiepileptic drugs(AEDs). The presence of p-glycoprotein and multi-drug resistance transporters in the blood-brain barrier could prevent the entry of AEDs into the brain, causing drug resistant epilepsy. To overcome this problem, we propose using carboxymethyl chitosan nanoparticles as a carrier to deliver carbamazepine(CBZ) intranasally with the purpose to bypass the blood-brain barrier thus to enhance the brain drug concentration and the treatment efficacy. Results so far indicate that the developed CBZNPs have small particle size(218.76 ± 2.41 nm) with high drug loading(around 35%) and high entrapment efficiency(around 80%). The in vitro release profiles of CBZ from the NPs are in accordance with the Korsmeyer-peppas model. The in vivo results show that both encapsulation of CBZ in nanoparticles and the nasal route determined the enhancement of the drug bioavailability and brain targeting characteristics.
基金Funded by Key Project of National Science Foundation of China (No.60537050)the National Science Foundation of China (No. 60377032)
文摘The magnetic chitosan nanoparticles were prepared by reversed-phase suspension method using Span-80 as an emulsifier, glutaraldehyde as cross-linking reagent. And the nanoparticles were characterized by TEM, FT-IR and hysteresis loop. The results show that the nanoparticles are spherical and almost superparamagnetic. The laccase was immobilized on nanoparticles by adsorption and subsequently by cross-linking with glutaraldehyde. The immobilization conditions and charac-terizations of the immobilized laccase were investigated. The optimal immobilization conditions were as follows: 10 mL of phosphate buffer (0.1 M, pH 7.0) containing 50 mg of magnetic chitosan nanoparticles, 1.0 mg·mL-1 of laccase and 1% (v/v) glutaraldehyde, immobilization temperature of 4 ℃ and immobilization time of 4 h. The immobilized laccase exhibited an appreciable catalytic capability (480 units·g-1 support) and had good storage stability and operation stability. The Km of immobilized and free laccase for ABTS were 140.6 and 31.1 μM in phosphate buffer (0.1 M, pH 3.0) at 37 ℃, respectively. The immobilized laccase is a good candidate for the research and development of biosensors based on laccase catalysis.
文摘pH-responsive-chitosan nanoparticles for the control release of protein drug were prepared by combining two-step crosslinking method, in which chitosan was subsequently crosslinked by sodium tripolyphosphate (TPP) and glycidoxypropyltrimethoxysilane (GPTMS). Compared with TPP crosslinked chitosan particles, the two-step crosslinked nanoparticles were not only pH-responsive but also more stable in wide pH range. Fluorescein isothiocyanate (FITC) labeled anti-human-IgG antibody was used as a model protein drug for evaluating the control release profile of the nano-carrier. The amount of released antibody increased from 5.6% to 50% when the pH of solution shifted from 7.4 to 6.0. The results suggest the possible application of the nanoparticles as pH- responsive drug delivery materials.
基金Shanghai Science and Technology Committee, No. 0452nm065
文摘AIM: To prepare chitosan-polyaspartic acid-5-fluorouracil (CTS-Pasp-5Fu) nanoparticles and investigate its anti-carcinoma effect and toxicity. METHODS: CTS-Pasp-5Fu nanoparticles were synthesized by ionic gelatification. Male BABL/c nude mice were injected with SGC-7901 gastric carcinoma cell line mass to establish a human gastric carcinoma model. They were randomly allocated into 4 groups: CTS-Pasp-5Fu (containing 5-Fu 1.25 mg/kg), 5-Fu (1.25 mg/kg), CTS-Pasp and normal saline groups. Tumor weight was measured and assay of colony forming unit-granulocyte and macrophage (CFU-GM) was performed. The structural change of cells and tissues was observed and the Bax and Bcl-2 genes were detected. RESULTS: Compared with normal saline, the inhibition rates of tumor growth for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.58%, 58.69% and 70.82%, respectively. The tumor inhibition rates for the CTS-Pasp, 5-Fu and CTS-Pasp-5Fu groups were 5.09%, 65.3% and 72.79%, respectively. There was a significant decrease in the number of CFU-GMformation and increase of total bilirubin, and alanine aminotransferase in the 5-Fu group, but no change in those of the other three groups. There was no change in white blood cell count and creatinine among the four groups. Pathological section of liver and nephridial tissues showed that the damage in the 5-Fu group was more severe than that in the CTS-Pasp-5Fu group. 5-Fu and CTS-Pasp-5Fu groups could both down-regulate the Bcl-2 expression and up-regulate the Bax expression to different extent, and the accommodate effect of CTS-Pasp-5Fu was more obvious than 5-Fu. CONCLUSION: The tumor inhibition rate of CTS-Pasp-5Fu nanoparticles is much higher than that of 5-Fu alone.
基金supported by the Anhui Provincial Natural Science Foundation (grant number 1508085QH194)
文摘The sustained-release properties of the biodegradable nano-drug delivery systems were used to improve the residence time of the chemotherapeutic agent in the body. These drug delivery systems were widely used to deliver chemotherapeutic drugs. The 5-fluorouracil loaded chitosan nanoparticles prepared in this paper have the above advantage. Here, we found that when the mass ratio of 5-fluorouracil and chitosan was 1:1, the maximum drug loading of nanoparticles was 20.13 ± 0.007%, the encapsulation efficiency was 44.28 ± 1.69%, the particle size was 283.9 ± 5.25 nm and the zeta potential was 45.3 ± 3.23 mV. The prepared nanoparticles had both burst-release and sustained-release phases in vitro release studies.In addition, the inhibitory effect of the prepared nanoparticles on gastric cancer SGC-7901 cells was similar to that of 5-fluorouracil injection, and the blank vector had no obvious inhibitory effect on SGC-7901 cells. In the pharmacokinetic study of rats in vivo, we found that AUC(0-t), MRT(0-t) and t1/2 z of nanoparticles were significantly increased in vivo compared with 5-fluorouracil solution, indicating that the prepared nanoparticles can play a role in sustained-release.
基金National Natural Science Foundation of China(30370344)Korea Science and Engineering Foundation(19992-220-009-4)supported this study
文摘Chitosan was modified by conjugating coupling with linolenic acid through the 1-ethyl-3-(3-dimethylami- nopropyyl) carbodiimide (EDC)-mediated reaction. The degree of substitution 1.8% ( i.e. 1.8 linolenic acid group per 100 anhydroglucose units) was measured by ^1H NMR. The critical aggregation concentration (CAC) of the self-aggregate of hydrophobically modified chitosan was determined by measuring the fluorescence intensity of the pyrene as a fluorescent probe. The CAC value in phosphate-buffered saline (PBS) solution (pH 7.4) was 5 × 10^-2 mg mL^-1. The average particle size of selfaggregates of hydrophobically modified chitosan in PBS solution (pH7.4) was 210.8 nm with a unimodal size distribution ranging from 100 to 500 nm. Transmission electron microscopy (TEM) study showed that the formation of near spherical shape nanoparticles has enough structural integrity. The loading ability of hydrophibically modified chitosan (LA-chitosan) was investigated by using bovine serum albumin (BSA) as the model. The loading capacity of self-aggregated nanoparticles increases ( 19.85 % ± 0.04 % to 37.57 % ± 0.25 % ) with the concentration of BSA (0.1-0.5 mg mL^-1 ).
基金supported by the Academic Research Fund,Faculty of Science,National University of Singapore,R148-000-180-112
文摘The aim of this paper is to investigate and optimize the preparation of scutellarin(SCU)-loaded HP-β-CD/chitosan(CS) nanoparticles(CD/CS-SCU-NPs). CD/CS-SCU-NPs were prepared by ionic cross-linking method and the process and formulation variables were optimized using response surface methodology(RSM) with a three-level, three factor Box–Behnken design(BBD).The independent variables were the added amounts of CS, sodium tripolyphosphate(TPP)and Pluronic F-68 during the preparation. Dependent variables(responses) were particle size and entrapment efficiency. Mathematical equations and respond surface plots were used to correlate independent and dependent variables.The preparation process and formulation variables were optimized to achieve minimum particle size and maximum entrapment efficiency by calculating the overall desirability value(OD). The optimized NP formulation was characterized for particle size, PDI, zeta potential, entrapment efficiency and in vitro drug release.According to the results, an optimized CD/CS-SCU-NP formulation was prepared. Results for particle size, PDI, zeta potential and entrapment efficiency were found to be around 200 nm,0.5, 25 mV, and 70% respectively. For in vitro study, the release of SCU from the NPs exhibited a biphasic release and was in accordance with Higuchi equation. The optimized preparation was simple with the probability for industrialization. The combination use of RSM, BBD and overall desirability values could provide a promising application for incorporating CD into CS nanoparticles as drug delivery carrier and help develop lab-scale procedures.
文摘AIM: To investigate the effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 in vitro and the possible mechanisms involved. METHODS: Chitosan nanoparticles were characterized by particle size, zeta potential, and morphology. After treatment with various concentrations of chitosan nanoparticles (25, 50, 75, 100 μg/mL) at various time intervals, cell proliferation, ultrastructural changes, DNA fragmentation, mitochondrial membrane potential (MMP), cell cycle phase distribution and apoptotic peaks of MGC803 cells were analyzed by MTT assay, electron microscopy, DNA agarose gel electrophoresis, and flow cytometry. RESULTS: Chitosan nanoparticles exhibited a small particle size as 65 nm and a high surface charge as 52 mV. Chitosan nanoparticles markedly inhibited cell proliferation of MGC803 cells with an ICso value of 5.3 μg/mL 48 h after treatment. After treatment with chitosan nanoparticles, the typical necrotic cell morphology was observed by electron microscopy, a typical DNA degradation associated with necrosis was determined by DNA agarose electrophoresis. Flow cytometry showed the loss of MMP and occurrence of apoptosis in chitosan nanoparticles-treated cells. CONCLUSION: Chitosan nanoparticles effectively inhibit the proliferation of human gastric carcinoma cell line MGC803 in vitro through multiple mechanisms, and may be a beneficial agent against human carcinoma.
基金supported by the National Science Foundation of China (No.21577037)Shanghai Committee of Science and Technology (No.17ZR1406600),Shanghai Committee of Science and Technology (grant No.11DZ2260600)Science and Technology Commission of Shanghai Municipality (STCSM,contract No.10DZ2220500)
文摘β-cyclodextrin (CD) grafted N-maleoyl chitosan (CD-g-NMCS) with two different degrees of substitution (DS) of N-maleoyl (DS = 21.2% and 30.5%) were synthesized from maleic anhydride and chitosan bearing pendant cyclodextrin (CD-g-CS). CD-g-NMCS based nanoparticles were prepared via an ionic gelation method together with chitosan and CD-g-CS nanoparticles.The size and zeta potential of prepared CD-g-NMCS nanoparticles were 179.2~274.0 nm and 36.2~42.4 m V, respectively. In vitro stability test indicated that CD-g-NMCS nanoparticles were more stable in phosphate-buffered saline compared with chitosan nanoparticles. Moreover, a poorly water-soluble drug, ketoprofen (KTP), was selected as a model drug to study the obtained nanoparticle’s potentials as drug delivery carriers. The drug loading efficiency of CD-g-NMCS20 nanoparticles were 14.8% for KTP. MTT assay showed that KTP loaded CD-g-NMCS nanoparticles were safe drug carriers. Notably, in vitro drug release studies showed that KTP was released in a sustained-release manner for the nanoparticles. The pharmacokinetic of drug loaded CD-g-NMCS20 nanoparticles were evaluated in rats after intravenous administration. The results of studies revealed that, compared with free KTP, KTP loaded CD-g-NMCS20 nanoparticles exhibited a significant increase in AUC0→24h and mean residence time by 6.6-fold and 2.9-fold, respectively. Therefore, CD-g-NMCS nanoparticles could be used as a novel promising nanoparticle-based drug delivery system for sustained release of poorly water-soluble drugs. The carboxylic acid groups of the CD-g-NMCS molecule provide convenient sites for further structural modifications including introduction of tissue-or disease-specific targeting groups.
文摘Objective To develop a safe and novel immunoadjuvant to enhance the immunity and resistance of animals against E. coli infection. Methods An 88-base immunostimulatory oligodeoxynuleotide containing eleven CpG motifs (CpG ODN) was synthesized and amplified by PCR. The chitosan nanoparticle (CNP) was prepared by ion linking method to entrap the CpG ODN that significantly promotes the proliferation of lymphocytes of pig in vitro. Then the CpG- CNP was inoculated into 21-day old Kunming mice, which were orally challenged with virulent K88/K99 E. Coil 35 days after inoculation. Blood was collected from the tail vein of mice on days 0, 7, 14, 21, 28, 35, 42, and 49 after inoculation to detect the changes and content of immunoglobulins, cytokines and immune cells by ELISA, such as IgG, IgA, IgM, IL-2, IL-4, and IL-6. Results The CpG provoked remarkable proliferation of lymphocytes of pig in vitro in comparison with that of control group (P〈0.05). The inoculation with CpG-CNP significantly raised the content of IgG, IgM, and IgA in the sera of immunized mice (P〈0.05). The levels of IL-2, 1L-4, and IL-6 in the mice significantly increased in comparison with those in controls (P〈0.05), so was the number of white blood cells and lymphocytes in immunized mice. The humoral and cellular immunities were significantly enhanced in immunized mice, which resisted the infection of E coli and survived, while the control mice manifested evident symptoms and lesions of infection. Conclusions CpG-CNP can significantly promote cellular and humoral immunity and resistance of mice against E. coli infection, and can be utilized as an effective adjuvant to improve the immunoprotection and resistance of porcine against infectious disease.
基金This work was supported by the 3rd Jiangsu Overseas Research&Training Program for University Prominent Young&Middleaged Teachers and Presidentsthe College Students Innovation Project for the R&D of Novel Drugs[No.J1310032]And we would like to thank cell and molecular biology experiment platform of China Pharmaceutical University for the assistance with relevant test items.
文摘Gene-based therapeutics has emerged as a promising approach for human cancer therapy. Among a variety of non-viral vectors, polymer vectors are particularly attractive due to their safety and multivalent groups on their surface. This study focuses on guanidinylated O-carboxymethyl chitosan(GOCMCS) along with poly-β-amino ester(PBAE) for si RNA delivery. Binding efficiency of PBAE/si RNA/GOCMCS nanoparticles were characterized by gel electrophoresis. The si RNA-loaded nanoparticles were found to be stable in the presence of RNase A, serum and BALF respectively. Fine particle fraction(FPF) which was determined by a two-stage impinger(TSI) was 57.8% ± 2.6%. The particle size and zeta potential of the nanoparticles were 153.8 ± 12.54 nm and + 12.2 ± 4.94 m V. In vitro cell transfection studies were carried out with A549 cells. The cellular uptake was significantly increased. When the cells were incubated with si Survivin-loaded nanoparticles, it could induce 26.83% ± 0.59% apoptosis of A549 cells and the gene silencing level of survivin expression in A549 cells were 30.93% ± 2.27%. The results suggested that PBAE/GOCMCS nanoparticle was a very promising gene delivery carrier.
文摘Chitosan is a natural polysaccharide prepared by the N-deacetylation of chitin. In this paper we have reviewed the methods of preparation of chitosan-based nanoparticles and their pharmaceutical applications. There are five methods of their prepa-rations:emulsion cross-linking,emulsion-droplet coalescence,ionic gelation,reverse micellar method and chemically modified chi-tosan method. Chitosan nanoparticles are used as carriers for low molecular weight drug,vaccines and DNA. Releasing characteris-tics,biodistribution and applications are also summarized.
基金Universiti Teknologi MARAMinistry of Higher Education of Malaysia for fund(0141903),LRGS-NanoMITe RU029-2014 and facility support。
文摘Chitosan nanoparticles are exhalation prone and agglomerative to pulmonary inhalation.Blending nanoparticles with lactose microparticles(~5 μm) could mutually reduce their agglomeration through surface adsorption phenomenon. The chitosan nanoparticles of varying size, size distribution, zeta potential, crystallinity, shape and surface roughness were prepared by spray drying technique as a function of chitosan, surfactant and processing conditions. Lactose-polyethylene glycol 3000(PEG3000) microparticles were similarly prepared. The chitosan nanoparticles, physically blended with fine lactose-PEG3000 microparticles, exhibited a comparable inhalation performance with the commercial dry powder inhaler products(fine particle fraction between 20% and 30%). Cascade impactor analysis indicated that the aerosolization and inhalation performance of chitosan nanoparticles was promoted by their higher zeta potential and circularity, and larger size attributes of which led to reduced inter-nanoparticulate aggregation and favored nanoparticles interacting with lactose-PEG3000 micropaticles that aided their delivery into deep and peripheral lungs.
基金National Natural Science Foundations of China(Nos.31470941,31271035)Science and Technology Commissions of Shanghai Municipality,China(Nos.15JC1490100,15441905100)+3 种基金Ph.D.Programs Foundation of Ministry of Education of China(No.20130075110005)Light of Textile Project,China(No.J201404)Yantai Double Hundred Talent Plan,China(No.XY-04-16-06)“111 Project”Biomedical Textile Materials Science and Technology,China(No.B07024)
文摘Chitosan(CS)nanofibers containing silver nanoparticles(AgNPs)were prepared by in-situ reducing method.A water soluble carboxymethyl chitosan(CMCT)was applied for the preparation of AgNPs.The impact factor such as the concentration of CMCT,silver nitrate(AgNO_3)content,temperature and the heating time during the preparation of AgNPs were studied.The result showed that the proper value of the concentration of CMCT,AgNO_3content,temperature and the heating time were set as0.1%,20μL AgNO_3(1.7 mol/L),90°and 3 h,separately and the maximum concentration of AgNPs could be acquired.To solve the spinnability of chitosan nanofiber,a super high molecular weight polyethylene oxide(PEO)was introduced to the system,and a new mixed solvent system was prepared by adding acetic acid,dimethyl sulfoxide(DMSO)and several drops of Triton X-100TMto distilled water.CS/PEO(80/20)with the concentration of 3%was dissolved in the mixed solvent to prepare electrospinning solution for CS/PEO(80/20)nanofiber fabrication.The CS containing AgNPs electrospun solution could be prepared by replacing the distilled water to silver nanoparticle solution during the preparation of mixed solvent.Ultraviolet visible(UV-Vis)spectra and transmission electron microscope(TEM)results showed that silver nanoparticles were prepared successfully.CS membranes with and without AgNPs were acquired via a traditional electrospinning equipment.These two nanofiber membranes were characterized by scanning electron microscope(SEM)images and mechanical testing.It could be noticed from the SEM images that there was a good morphology and random distribution for the nanofibers with an average fiber diameter of 180 nm.The mechanical property results showed that the addition of AgNPs decreased the mechanical strength significantly but the mechanical strength could still support wound dressing application.
文摘AIM:To study the expression of human insulin gene in gastrointestinal tracts of diabetic rats. METHODS: pCMV.Ins, an expression plasmid of the human insulin gene, wrapped with chitosan nanoparticles, was transfected to the diabetic rats through lavage and coloclysis, respectively. Fasting blood glucose and plasma insulin levels were measured for 7 d. Reverse transcription polymerase chain reaction (RT-PCR) analysis and Western blot analysis were performed to confirm the expression of human insulin gene. RESULTS: Compared with the control group, the fasting blood glucose levels in the lavage and coloclysis groups were decreased significantly in 4 d (5.63 ± 0.48 mmol/L and 5.07 ± 0.37 mmol/L vs 22.12 ± 1.31 mmol/L, respectively, P < 0.01), while the plasma insulin levels were much higher (32.26 ± 1.81 μIU/mL and 32.79 ± 1.84 μIU/mL vs 14.23 ± 1.38 μIU/mL, respectively, P < 0.01). The human insulin gene mRNA and human insulin were only detected in the lavage and coloclysis groups. CONCLUSION: Human insulin gene wrapped with chitosan nanoparticles can be successfully transfected to rats through gastrointestinal tract, indicating that chitosan is a promising non-viral vector.
基金Projects(31201074,81371013) supported by the National Natural Science Foundation of ChinaProject(2011105102016) supported by the Key Program of Medical Health of Dongguan City,Guangdong Province,ChinaProject(2011108102026) supported by Dongguan Universities Program,China
文摘Folic acid conjugated chitosan was prepared by cross-linking reaction with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC), and then used as a template to prepare folic acid-chitosan(FA-CS) conjugated nanoparticles and load mitoxantrone nanoparticles(FA-CSNP/MTX). Drug dissolution testing, CCK-8 method, and confocal microscopy were used to detect their controlled-release capability in different situations and the specific uptake by HONE1 cells. The experimental results show that the nanoparticles have uniform size distribution of 48-58 nm. The highest encapsulation rate of the particles on mitoxantrone hydrochloride(MTX) is(77.5±1.9)%, and the drug loading efficiency is(18.4±0.4)%. The sustained release effect, cell growth inhibition activity and targeting effect of the FA-CS/MTX nanoparticles are good in artificial gastric fluid and intestinal fluid. It is demonstrated that the FA-CSNP system is a potentially useful system for the targeted delivery of anticancer drug MTX.