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Background: Irrigation is a crucial aspect of root canal treatment, and it is imperative to employ chelating agents to eliminate the smear layer during biomechanical preparation. They in turn react with mineral content of dentin, leading to decreased strength and increased susceptibility to fracture. Aims: This study aimed to assess and compare mineral loss and microhardness from primary root canal dentin following the usage of different irrigating solutions and determine the least detrimental irrigant among the tested solutions. Materials and Methods: Sixty-six primary anterior teeth were divided into three groups with 22 in each– Group I: 17% ethylenediaminetetraacetic acid (EDTA) Group II: 0.2% Nano chitosan Group III: Pomegranate extract. The decoronated teeth were split longitudinally. Half of it was directly subjected to Vickers test, and the other half was immersed in a magnetic stirrer bath containing test solution to record the mineral loss from solution. Postmicrohardness values were recorded on the specimen and compared with initial values. Statistical Analysis Used: Statistical analysis was done using SPSS software (Version 20, SPSS, IBM, Armonk, NY, U. S. A). Results: Descriptive statistics were calculated, and the groups were compared using analysis of variance test and post hoc Tukey test. Pomegranate extract showed least effect on mineral content and microhardness compared to 17% EDTA and 0.2% nano chitosan. P ≤ 0.05 was considered statistically significant. Conclusion: Pomegranate aril extract showed better results with selected parameters.
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@#The present study compares the in vitro effects of nanoparticles loaded pentamidine drug and conventional pentamidine on Leishmania tropica. Herein, pentamidine-loaded chitosan nanoparticles (PTN-CNPs) have been synthesized through an ionic gelation method with sodium tripolyphosphate (TPP). Next, the physical characteristics of PTN-CNPs were determined through the surface texture, zeta potential, in vitro drug release, drug loading content (DLC), and encapsulation efficacy (EE) and compared its efficacy with free pentamidine (PTN) drug against promastigotes and axenic amastigotes forms of L. tropica in vitro. The PTN-CNPs displayed a spherical shape having a size of 88 nm, an almost negative surface charge (-3.09 mV), EE for PTN entrapment of 86%, and in vitro drug release of 92% after 36 h. In vitro antileishmanial activity of PTN-CNPs and free PTN was performed against Leishmania tropica KWH23 promastigote and axenic amastigote using 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyletetrazolium bromide (MTT) assay. It was observed that the effect of PTN-CNPs and free PTN on both forms of the parasite was dose and time dependent. Free PTN presented low efficacy even at higher dose (40 µg/ml) with 25.6 ± 1.3 and 26.5 ±1.4 mean viability rate of the promastigotes and axenic amastigotes, respectively after 72 hrs incubation. While PTN-CNPs showed strong antileishmanial effects on both forms of parasite with 16 ± 0.4 and 19 ± 0.7 mean viability rate at the same higher concentration (40 µg/ml) after 72 hrs incubation. Half maximal inhibitory concentration (IC50) values of PTN-CNPs toward promastigotes and amastigotes were obtained as 0.1375 µg/ml and 0.1910 µg/ml, respectively. In conclusion, PTN-CNPs effectively inhibited both forms of the L. tropica; however, its effect was more salient on promastigotes. This data indicates that the PTN-CNPs act as a target drug delivery system. However, further research is needed to support its efficacy in animal and human CL.
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Aims@#This study was designed to evaluate the effectiveness of the synthesised carvacrol loaded chitosan nanoparticles (CLCNPs) on the growing and pre-formed biofilms of Listeria monocytogenes isolated from slaughterhouses.@*Methodology and results@#The swab samples were collected from knives, hocks and cutting tables representing slaughterhouses meat contact surfaces (MCS), while those samples from walls and floors represent slaughterhouses meat non-contact surfaces (MNCS). The bacteriological analysis revealed the existence of L. monocytogenes with a prevalence rate of 3.3, 10 and 6.7% for knives, hocks and cutting tables, respectively and 2.2 and 6.6% for walls and floors, respectively. The isolates L. monocytogenes were assayed for biofilm production by the crystal violet binding assay method. Among the 10 L. monocytogenes isolates, 10%, 50% and 30% of the isolates were found to be strong, moderate and weak biofilm producers, respectively. The activities of carvacrol, chitosan nanoparticles (NPs) and CLCNPs against the only strong biofilm producer strain of L. monocytogenes were tested by microtiter plate assay. The minimum inhibitory concentrations (MIC) values were 3.75 mg/mL for CAR, 5 mg/mL for chitosan NPs and 0.62 mg/mL for CLCNPs. CLCNPs inhibit the produced biofilm by 35.79, 73.37 and 77.76%, when 0.5 MIC, 1 MIC and 2 MIC were used, respectively. Furthermore, the pre-formed L. monocytogenes biofilms were significantly reduced from 1.01 (control) OD570 to 0.40 and 0.29 OD570 by applying 2 MIC and 4 MIC doses, respectively.@*Conclusion, significance and impact of study@#The data generated is promising to develop bio-green disinfectants to inhibit biofilm formation by L. monocytogenes in the food processing environment and control its adverse effects for consumers.
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Listeria monocytogenes , NanoparticlesABSTRACT
Objective: The purpose of this study was to evaluate the effect of chitosan nanoparticles on microtensile bond strength of resin composite to dentin using self etch adhesive after aging. Material and Methods: A total number of 90 freshly extracted, sound human molar teeth. Flat tooth surface was gained after cut of the occlusal surface. Three main groups according to pretreatment of dentin before adhesive application; 0.2 % chitosan, 2.5 % chitosan and no treatment control group. Universal self etch adhesive were applied according to manufacture instruction and 4 mm of Feltik Z250 xt composite. Storage of specimens for 1 day, 3 months and 6 months in 37O C distilled water. After that, the tooth was sectioned to beams of 1 mm x8 mm sticks for microtensile bond strength test using universal testing machine. Scanning electron microscope (SEM) was used to evalute the effect of chitosan nanoparticles on dentin and smear layer. Kruskal-Wallis test was used to compare between the three groups as well as the three aging periods. Dunn's test was used for pair-wise comparisons. The significance level was set at P ≤ 0.05. Results: chitosan 0.2% is statistically significant increase in bond strength than chitosan 2.5% and control in one day group. Three months chitosan 0.2 % groups have statistically significant increase in bond strength than chitosan 2.5%. It was found in 6 months that control and chitosan 0.2 % have statistically significant increase in bond strength than chitosan 2.5%. There was statistically significant difference found between the three studied groups regarding bond strength at different storage times . Conclusion: Microtensile bond strength was influenced by different chitosan concentration. Different aging periods had no effect on the microtensile bond strength without application of chitosan and with application of 2.5% chitosan concentration. (AU)
Introdução: O objetivo deste estudo foi avaliar o efeito das nanopartículas de quitosana na resistência da microtração de união do compósito de resina à dentina usando adesivo autocondicionante após o envelhecimento. Material e Métodos: Foram utilizados um total de 90 dentes molares humanos extraídos e sadios. A superfície plana do dente foi obtida após o corte da superfície oclusal. Os dentes foram divididos em três grupos principais de acordo com o pré-tratamento da dentina e antes da aplicação do adesivo: 0,2% de quitosana, 2,5% de quitosana e nenhum tratamento foi utilizado no grupo controle. O adesivo autocondicionante universal foi aplicado de acordo com as instruções do fabricante e 4 mm de composito Feltik Z250 xt foi inserido. O armazenamento de amostras foi realizado por 1 dia, 3 meses e 6 meses em água destilada a 37 °C. Depois disso, o dente foi seccionado em peças de 1 mm x 8 mm para teste de resistência de união por microtração, utilizando máquina de teste universal. Microscópio eletrônico de varredura (MEV) foi usado para avaliar o efeito das nanopartículas de quitosana na dentina e na camada de smear layer. O teste de Kruskal-Wallis foi utilizado para comparar os três grupos e os três períodos de envelhecimento. O teste de Dunn foi usado para comparação pareada dos grupos. O nível de significância foi estabelecido em P ≤ 0,05. (AU)
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Humans , Matrix Metalloproteinases , Dentin , Chitosan , MolarABSTRACT
Objective: To evaluate the effect of low molecular weight chitosan (LMW-CTS) and its nanoparticles (LMW-CTS-NPs) on the intestinal permeability of Panax notoginseng saponins (PNS) by using Caco-2 cell model. Methods: LMW-CTS was prepared by combining chitosanase hydrolysis combined with ultrafiltration separation technology, and molecular weight of LMW-CTS was determined by using permeation gel chromatography (GPC). LMW-CTS-NPs were prepared by ionic gel method, and characterized by scanning electron microscopy, nano particle sizer, and flourier transformation infrared spectroscopy. Caco-2 cell model was established and validated to evaluate the effects of LMW-CTS and LMW-CTS-NPs on the intestinal permeability of PNS. Results: LMW-CTS has a molecular weight of 5 760 and a polydispersity coefficient of 1.42. LMW-CTS-NPs have a round shape and narrow particle size distribution, with an average particle size of 115.5 nm and zeta potential of +37.1 mV. The apparent permeability coefficients (Papp, AB→BL) of PNS was less than 1 × 10-6 cm/s, indicating a poor permeability. In LMW-CTS group, the Papp of R1 and Rg1 was increased by 17.83% and 20.29%, respectively, but no significant effect of promotion was observed on other components. However, the Papp of R1, Rg1, Re, Rb1, and Rd in LMW-CTS-NPs group was increased by 35.66%, 23.28%, 29.41%, 37.99%, and 36.00%, respectively, compared tothe control group. Conclusion: LMW-CTS can significantly promote the intestinal mucosal permeability of R1 and Rg1 in PNS, but has no significant effect on Re, Rb1, and Rd. LMW-CTS-NPs significantly increased the permeability of the major monomer saponin components in PNS. Namely, the intestinal permeability of PNS can be further improved by transforming LMW-CTS into LMW-CTS-NPs.
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PURPOSE: Escherichia coli O157:H7 is one of the most important pathogens which create hemorrhagic colitis and hemolytic uremic syndrome in human. It is one of the most prevalent causes of diarrhea leading to death of many people every year. The first diagnosed gene in the locus of enterocyte effacement pathogenicity island is eae gene. The product of this gene is a binding protein called intimin belonging to the group of external membrane proteins regarded as a good stimulants of the immune system. Chitosan with its lipophilic property is an environmentally friendly agent able to return to the environment. MATERIALS AND METHODS: Intimin recombinant protein was expressed in pET28a vector with eae gene and purification was performed using Ni-NTA and finally the recombinant protein was approved through western blotting. This protein was encapsulated using chitosan nanoparticles and the size of nanoparticles was measured by Zetasizer. Intimin encapsulated was prescribed for three sessions among three groups of oral, injection, and oral-injection using Chitosan nanoparticles. Challenge was performed for all three groups with 108 E. coli O157:H7 bacteria. RESULTS: Intimin produced by chitosan nanoparticles improves immunological responses through the adjuvant nature of chitosan nanoparticles. Chitosan may be used as a carrier for transportation of the prescribed vaccine. Among the mice, encapsulated intimin could be able to provide suitable titers of IgG and IgA by the aid of chitosan nanoparticles. Results of mice challenge showed that decreased the bacterial shedding significantly. CONCLUSION: Results showed that the chitosan nanovaccine with intimin protein may be used as a suitable candidate vaccine against E. coli O157:H7.
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Animals , Humans , Mice , Bacteria , Bacterial Shedding , Blotting, Western , Carrier Proteins , Chitosan , Colitis , Diarrhea , Enterocytes , Escherichia coli , Genomic Islands , Hemolytic-Uremic Syndrome , Immune System , Immunoglobulin A , Immunoglobulin G , Membrane Proteins , Nanoparticles , TransportationABSTRACT
In this study, an attempt was made to develop bi-functional constructs serving both as scaffolds and potential delivery systems for application in neural tissue engineering. The constructs were prepared in two steps. In the first step, the bulks of poly (L-lactic acid) (PLLA) in 1, 4-dioxane/water (87:13) were fabricated using liquid-liquid thermally induced phase separation technique. In the next step, the prepared bulks were coated with chitosan nanoparticles produced by two different techniques of ultrasonication and ionic gelation by grafting-coating technique. In ultrasonication technique, the chitosan solution (2 mg/mL) in acetic acid/sodium acetate buffer (90:10) was irradiated by an ultrasound generator at 20 kHz and power output of 750 W for 100 s. In ionic gelation technique, the tripolyphosphate in water solution (1 mg/mL) was added to the same chitosan solution. The physicochemical properties of the products were characterized by Scanning Electron Microscopy, Attenuated Total Reflection Fourier Transform-Infrared, liquid displacement technique, contact angle measurement, compressive and tensile tests, as well as zeta potential and particle size analysis using dynamic light scattering. Moreover, the cell proliferation and attachment on the scaffolds were evaluated through human glioblastoma cell line (U-87 MG) and human neuroblastoma cell line [BE (2)-C] culture respectively. The results showed that the samples coated with chitosan nanoparticles prepared by ultrasonication possessed enhanced hydrophilicity, biodegradation and cytocompatibility compared with pure PLLA and PLLA coated with chitosan nanoparticles prepared by ionic gelation. This study suggests successful nanoparticles-scaffold systems which can act simultaneously as potential delivery systems and tissue engineering scaffolds.
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Humans , Cell Line , Cell Proliferation , Chitosan , Dynamic Light Scattering , Glioblastoma , Hydrophobic and Hydrophilic Interactions , Microscopy, Electron, Scanning , Nanoparticles , Neuroblastoma , Particle Size , Tissue Engineering , Ultrasonography , WaterABSTRACT
Objective: To prepare the epigallocatechin-3-gallate (EGCG) chitosan nanoparticles (CS-NPs) and investigate their physicochemical properties. Methods: The EGCG-CS-NPs were prepared by ion gelation method. The formulation variables were optimized by Box-Behnken Design (BBD) of response surface methodology (RSM) of CS concentration (X1), sodium tripolyphosphate concentration (X2), and EGCG concentration (X3) as independent variables and encapsulation efficiency (Y1,%) and particle size (Y2, nm) as dependent variables. The optimized CS-NPs were characterized for encapsulation efficiency (EE), particle size, Zeta potential, morphology, and in vitro drug release behavior of EGCG-CS-NPs were studied. Results: An optimal EGCG-CS-NPs consisting of CS concentration as 2.6 g/L, sodium tripolyphosphate concentration as 1.5 g/L and EGCG concentration as 2.7 g/L. For EE, particle size, Zeta potential of EGCG-CS-NPs were found to be (85.8±3.1)%, (102.2±27.1) nm, and (25.5±4.1) mV, respectively. The CS-NPs were found to be small and spherical as seen in transmission electron microscopy (TEM). The in vitro release data proved that the drug release was steady within 24 h (pH 4.5 PBS). Conclusion: Through optimizing the formulation, we obtain the uniform EGCG-CS-NPs with in vitro sustained-release behavior. This work is useful for the further research on pharmacodynamics of EGCG-CS-NPs.
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Objective: To investigate the effect of a method for imaging lung cancer cells using nanotechnology and molecular beacon (MB) that identifies miR-155 and is delivered by chitosan nanoparticles (CS). Methods: The miR-155 MB modified by locked nucleic acids (LNAs) was designed and synthesized. The CS-MB complex was synthesized by self-assembly method and tested for its physicochemical properties including anti-DNase I features, particle size, zeta potential and so on. The miR-155 MB was transfected with CS as vectors. The abilities of miR-155 MB to identify miR-155 and thus to image lung cancer cells were determined by confocal microscopy. Furthermore, the miR-155 expression levels were detected by qRT-PCR to validated the effect of miR-155 MB. The random sequence molecular beacon (RS MB) was set as a negative control. Results: The CS-MB complex at the weight ratio of 7:1 was best suited for transfection due to its high encapsulation rate, resistance to the degradation by DNase I, small particle size and positive charge. Relatively strong red fluorescence could be detected in the lung cancer cells after transfection of miR155 MB, while that could not be detected in the RS MB group (P<0.05). Moreover, the changing trend in the fluorescence intensity was consistent with that in the miR-155 expression levels. Conclusion: CS nanoparticles can be used as vectors to deliver miR-155 MB for miR-155 identification and lung cancer cell imaging, thus providing new ideas and novel technique for lung cancer diagnosis.
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Herein we describe the preparation, characterization and utilization of chitosan nanoparticles for the intracellular delivery of the poorly cell-penetrating antibiotic e.g. Ciprofloxacin, Chlortetracycline hydrochloride and Gentamycin sulfate to improve their treatment of bacterial infections. Chitosan nanoparticles were prepared via the ionic gelation of chitosan with tri polyphosphate anions. Several parameters were studied to optimize the particle size of chitosan nanoparticles, here we select the concentration of chitosan and the concentrations of sodium tri poly phosphate (TPP) as optimizing parameters and the other factors stay constant such as pH of solution and ultrasonication time. Chitosan nanoparticles formed characterized by using FT-IR and transmission electron microscope (TEM). Results show that chitosan nanoparticles and its loaded antibiotics kill and inhibits the growth of gram (+) and gram (-) bacteria tested due to nanoparticles structures, and the antibacterial activity increased with increasing the anti biotic content.
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Objective To prepare self-assembled thiolated chitosan derivatives gold nanoparticles (CS-GNRs) and carry out the feature tests.Methods CS-GNRs was prepared by chitosan derivatives and GNRs through strong metal sulfur chemical bond between thiols and gold on GNRs surface.Morphology features was tested by transmission electron microscope,dynamic light scattering was adopted to observe the size of nanoparticles.Ultraviolet-visible spectrophotometer was used to detect the optical properties and the property change.Meanwhile,the surface-enhanced Raman scattering (SERS) activity of CS-GNRs was investigated by using crystal violet (CV) as a probe molecular.Results CS-GNRs were in good shape,uniform particle size and good dispersion.The SERS of CV was enhanced,and the enhancement factor of CV adsorbed on CS-GNRs was up to 2×103.Conclusions The nanoparticles have potential application in molecular detection and Raman spectra detection.
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Objective To evaluate the internal irradiation biological effects of 125I-UdR chitosan nanoparticles in hepatoma cells.Methods The accumulation and distribution of 125I-UdR-CS-DLN in hepatoma cells HepG2 and human liver tissue cells HL-7702 were observed with a confocal microscopy.The internal irradiation biological effects were evaluated by MTT assay,flow cytometry and single cell gel electrophoresis.The apoptosis of in situ rabbit liver tumor treated with 125I-UdR-CS-DLN was assayed by TUNEL staining technique.Results After 30 min of nano-particle treatment,its accumulation in the cytoplasm of HepG2 cells was significantly greater than that in HL-7702 cells.When the concentrations of 125I-UdR-CS-DLN was higher than 37 kBq/ml,the cell viability of HepG2 was significantly lower than that of lL-7702 at 24 and 48 h post-treatment(t =-4.46-6.31,P＜0.05),and the HepG2 cells were arrested at G1 phase and significantly impaired at G2/M phase.In addition,the degrees of DNA doublestrand break of both cell lines irradiated by 125I-UdR-CS-DLN were significantly higher than those treated with 125I-UdR,and the DNA repair capacity of HepG2 cells was significantly lower than that of HL-7702 cells(OTM:t =2.94,P ＜0.05;TDNA％:t =10.64,P ＜0.01).TUNEL staining showed that cell apoptosis could be induced in the rabbit liver carcinoma by 125I-UdR-CS-DLN but not by 125I-UdR.Conclusions The amount of 125I-UdR-CS-DLN absorbed by hepatoma cells is significantly higher than that of 125I-UdR,which suggests that 125 I-UdR-CS-DLN induces more stronger internal radiation biological effects of apoptosis and DNA damage on hepatoma cells.
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The objective of the research was to formulate and evaluate selegiline hydrochloride loaded chitosan nanoparticles for the Parkinson's therapy in order to improve its therapeutic effect and reducing dosing frequency. Taguchi method of design of experiments (L9 orthogonal array) was used to get optimized formulation. The selegiline hydrochloride loaded chitosan nanoparticles (SHPs) were prepared by ionic gelation of chitosan with tripolyphosphate anions (TPP) and tween 80 as surfactant. The SHPs had a mean size of (303.39 ± 2.01) nm, a zeta potential of +32.50mV, and entrapment efficiency of SHPs was 86.200 ± 1.38%. The in vitro drug release of SHPs was evaluated in phosphate buffer saline (pH 5.5) using goat nasal mucosa and found to be 82.529% ± 1.308 up to 28 h. Release kinetics studies showed that the release of drug from nanoparticles was anomalous (non-fickian) diffusion indicating the drug release is controlled by more than one process i.e. superposition of both phenomenon, the diffusion controlled as well as swelling controlled release. SHPs showed good stability results as found during stability studies at different temperatures as mentioned in ICH guidelines. The results revealed that selegiline hydrochloride loaded chitosan nanoparticles are most suitable mode of delivery of drug for promising therapeutic action.
O objetivo da pesquisa foi formular e avaliar nanopartículas de quitosana contendo cloridrato de selegilina para terapia do Parkinson, a fim de melhorar o seu efeito terapêutico e reduzir a frequência de dosagem. Método de Taguchi, de planejamento experimental, (L9 matriz ortogonal) foi usado para obter a formulação otimizada. As nanopartículas de quitosana contendo cloridrato de selegilina (PCHs) foram preparadas por gelificação iônica de quitosana com ânions tripolifosfato (TPP) e Tween 80 como tensoativo. As PCHs apresentaram tamanho médio de (303.39 ± 2,01) nm, potencial zeta de +32.50 mV e eficiência de encapsulação de 86.200±1,38%. A liberação do fármaco in vitro foi avaliada em solução salina de tampão fosfato (pH 5,5), usando a mucosa nasal de cabra e o resultado encontrado foi de 82.529% ± 1.308, acima de 28 h. Estudos de cinética de liberação mostraram que a liberação do fármaco das nanopartículas foi por difusão anômala (não fickiana), indicando que é controlada por mais de um processo, ou seja, a superposição dos fenômenos de difusão controlada e intumescimento. As PCHs mostraram resultados de boa estabilidade, encontrada durante os estudos de estabilidade em temperaturas diferentes, como mencionado em diretrizes do ICH. Os resultados revelaram que o sistema de nanopartículas de quitosana contendo cloridrato de selegilina é o mais adequado sistema de liberação de fármacos de ação terapêutica promissora.
Subject(s)
Parkinson Disease/therapy , Nanoparticles , Selegiline/analysis , Chemistry, Pharmaceutical , Chitosan/analysis , Drug LiberationABSTRACT
Objective Using surface plasmon resonance (SPR)technology to monitor the interactions between chitosan nanoparticles (CS-NP)and biomembrane. Methods SPR signals induced by CS-NP,chitosan molecules and thymopentin(TP5) molecules in living C6 cells were detected and analyzed by the SPR-based cytosensor. Results The signal began to increase within 30 min after the addition of CS-NP or chitosan molecules. These responses were strongly dependent on the concentrations of CS. However,the signal did not change after the addition of TP5 molecules. Conclusion SPR technology can clarify the interaction between nanoparticles and biomembrane in real time.
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Objective To prepare and characterize nanoparticles in-loaded one-way release-controlled chitosan membrane,and to explore the release-controlled rule of the film in vitro.Methods The chitosan nanoparticles were prepared by inverse crosslinking-emulsion method.The one-way release-controlled membrane was prepared by a casting method.Transmission electron microscope (TEM) was used to evaluate the morphological properties and particle size analyzer was used to analyze particle size distribution.The morphology of the membrane was inspected through scanning electron microscope (SEM).MTT assay was applied to determine the biological safety of chitosan nanoparticles.The distribution of the nanoparticles was observed by fluorescence microscope.The in vitro release studies were adopted to evaluate the release-controlled rule.Results The four kinds of nanoparticles had spherical shapes and uniform particle size.The size of the hyaluronic acid-coated chitosan nanoparticle was (255.40±39.10) nm.Hyaluronic acid-coated chitosan nanoparticles showed the best property of sustained release and biocompatibility.The membrane had a loose inner layer and a dense outer layer,and the distribution of the nanoparticles was uniform in the inner layer of the membrane.The release of protein from membrane was unidirectional and the membrane displayed good controlled release property.Conclusions The nanoparticles in-loaded one-way release-controlled chitosan membrane presents good one-way sustained release performance.It is potentially useful in delivery system of growth factors.
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Objective: Using surface plasmon resonance(SPR) technology to monitor the interactions between chitosan nanoparticles(CS-NP) and biomembrane. Methods: SPR signals induced by CS-NP, chitosan molecules and thymopentin (TP5) molecules in living C6 cells were detected and analyzed by the SPR-based cytosensor. Results: The signal began to increase within 30 min after the addition of CS-NP or chitosan molecules. These responses were strongly dependent on the concentrations of CS. However, the signal did not change after the addition of TP5 molecules. Conclusion: SPR technology can clarify the interaction between nanoparticles and biomembrane in real time.
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Objective: To prepare salidroside-chitosan nanoparticles (SA-CS-NPs) and to evaluate the properties of in vitro drug release. Methods: SA-CS-NPs were firstly prepared by solvent diffusion-ionic crosslinking method. The particle size and polydispersity of SA-CS-NPs were determined and the morphology of nanoparticles was evaluated. The properties of encapsulation efficiency (EE), load efficiency (LE), and in vitro release of SA-CS-NPs were also evaluated using UPLC method. Results: The nanoparticles were successfully prepared with the spherical shape or para-spherical shape. The mean particle size was (247.5 ± 23.8) nm with the polydispersity index (PDI) of 0.265 ± 0.071, and the Zeta potential was (23.4 ± 2.7) mV (n = 3). The EE was (70.15 ± 1.60)% and the LE was (14.03 ± 0.32)% (n = 3). The cumulative release rate of SA-CS-NPs within 24 h was over 85%. Conclusion: SA-CS-NPs prepared by solvent diffusion-ionic crosslinking method show appropriate particle size and EE, and could exhibit sustained release properties in vitro.
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OBJECTIVE@#To explore the effect of sustained-release recombinant human bone morphogenetic protein-2 (rhBMP-2) on ectopic osteogenesis in the muscle pouches of rats through preparing rhBMP-2 sustained-release capsules by wrapping morphogenesis protein bones-2 (BMP-2) using chitosan nanoparticles, and compositing collagen materials.@*METHODS@#Twenty four Sprague-Dawley rats were randomly divided into four groups with six rats in each group, that is Group A (control group), Group B (only treated with collagen), Group C (rhBMP-2+collagen treated group) and Group D (rhBMP-2/cs+collagen treated group). The composite materials for each group were implanted in the bilateral peroneal muscle pouches in rats. The peroneal muscles were only separated without implanting any materials in control group. Rats were sacrificed 2 weeks and 4 weeks post treatment and samples were cut off for general observation, Micro CT scans and histological observation.@*RESULTS@#General observation showed no new bone formation in Groups A and B mice, while new bones were formed in Groups C and D mice. Two weeks after treatment Micro CT scans showed that The bone volume fraction (BVF), trabecular thickness (Tb.Th), bone mineral density (BMD) in Group C mice were all higher than that in Group D (P<0.05). At the fourth week, the BVF, Tb.Th and BMD were significantly higher than that at the second week (P<0.01).@*CONCLUSIONS@#The slow-release effect of rhBMP-2/cs sustained-release capsules can significantly promote ectopic osteogenesis. Its bone formation effect is better than that of rhBMP-2 burst-release group.
Subject(s)
Animals , Rats , Bone Morphogenetic Protein 2 , Pharmacology , Collagen , Pharmacology , Delayed-Action Preparations , Pharmacology , Drug Carriers , Pharmacology , Intercellular Signaling Peptides and Proteins , Muscle, Skeletal , Nanocapsules , Osteogenesis , Rats, Sprague-Dawley , Recombinant Proteins , Pharmacology , Tissue Engineering , Methods , Transforming Growth Factor beta , Pharmacology , X-Ray MicrotomographyABSTRACT
Objective To investigate the immune response of mucosal immunization of new chitosan(CS) nanoparticles coating DNA vaccine. Methods The chitosan nanoparticles containing plasmid DNA encoding H. pylori lipoprotein Lpp20 gene were prepared using a complex coacervation method and then its speciality were analyzed. We then administered the naked plasmid DNA and chitosan-DNA nanoparticles to 6-week-old female BALB/c mice by intranasal or oral mucosal routes to observe the humoral and cellular immune responses. Results Naked plasmid pcDNA3.1 ( + )/Lpp20 and chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles both induced effective immune response in mice through mucosal vaccination. Specific IgG and sIgA antibodies of chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles groups were higher than that of naked pcDNA3.1 ( + )/Lpp20 group. The concentration of cytokines IFN-γ and IL-4 in cultural supernatant of T lymphocytes from chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles immunized mice increased greatly than that of control groups. After stimulated by corresponding antigen, the stimulation index of intranasal or oral delivery of chitosan-pcDNA3. 1 ( + )/Lpp20 nanoparticles group was significantly higher than that of pcDNA3.1( + )/Lpp20 group, CS group and PBS control group. Moreover, systemic and mucosal immune responses in mice induced by intranasal immunization were stronger than that of oral immunization. Conclusion Chitosan nanoparticles enhanced the immune response of pcDNA3.1 ( + )/Lpp20 DNA vaccine by intranasal or oral administration in BALB/c mice. Compared to oral administration, intranasal delivery of chitosan-pcDNA3.1 ( + )/Lpp20 DNA nanoparticles could induce stronger cellular and humoral immune responses in BALB/c mice.
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OBJECTIVE: To investigate the anti-tumor activity of adriamycin-loaded chitosan nanoparticles surface-modified with glycyrrhizin on human hepatoma cell line SMMC-7721. METHODS: Inhibition effect of drug-loaded nanoparticle on the proliferation of SMMC-7721 cells was determined with MTT method. Intra-cellular distribution of nanoparticles and morphological change of SMMC-7721 cells before and after treatment were observed by confocal laser scanning microscope (CLSM) and the apoptosis induced by adriamycin was measured by TUNEL assay. RESULTS: The inhibition effect of adriamycin-loaded chitosan nanoparticles surface-modified with glycyrrhizin on the proliferation of SMMC-7721 cells increased 6.36 times lower in comparison with free drugs. The inhibition effect of adriamycin-loaded chitosan nanoparticles had no obvious improvement. Nanoparticles surface-modified with glycyrrhizin promoted the distrubition of adriamycin in the nucleus, so as to promote the anti-tumor activity of adriamycin. TUNEL assay indicated that nanoparticles surface-modified with glycyrrhizin induce DNA fragmentation and nucleus breakdown to induce the apoptosis of SMMC-7721 cells. CONCLUSION: Adriamycin-loaded chitosan nanoparticles surface-modified with glycyrrhizin can used as potential active hepatocyte-targeted delivery carrier. Pharmacokinetic evaluation of it is worthy of further studing.