ABSTRACT
Abstract Acne Vulgaris is a common skin disease caused by Propionibacterium acnes, an anaerobic microbiota of human skin that plays a vital role in the pathology of acne. The aim of this study was to prepare nanoparticles containing an acne recombinant protein and determine its ability as an oral acne vaccine in mice. The recombinant Sialidase-CAMP gene was expressed and purified in a prokaryotic host. The chitosan nanoparticles containing the recombinant protein were prepared, encapsulated, and administered by both oral and subcutaneous routes to Balb/c mice. Sera IgA and IgG and stool IgA titers were measured by ELISA, and the immunized mice were challenged against P. acnes. A 65 kDa recombinant protein was confirmed by SDS-PAGE and western blot. The size and zeta potential of nanoparticles were 80 nm and +18 mV, respectively. After oral immunization, the serum IgG and IgA titers were 1:3200 and 1:16, respectively, and the stool IgA titer was 1:8. In the subcutaneous route, the serum IgG titer was 1:51200. Immunized mice showed no inflammation in the ear of challenged mice. It is the first study that examines a chitosan-nanoparticulated acne fusion protein as an applicable acne vaccine candidate with appropriate immunogenicity potential. Further studies are required to validate the clinical usefulness of this vaccine candidate.
Subject(s)
Animals , Female , Mice , Propionibacterium acnes/drug effects , Acne Vulgaris/prevention & control , Chitosan/administration & dosage , Nanoparticles/administration & dosage , Recombinant Proteins , Enzyme-Linked Immunosorbent Assay , Blotting, Western , Immunization/methods , Disease Models, Animal , Electrophoresis, Polyacrylamide Gel , Mice, Inbred BALB C , NeuraminidaseABSTRACT
Objective To study and optimize the preparation condition of pVAX1‐wapA‐loaded nanoparticles and deter‐mine the transfection efficiency .Methods The related effects of the crucial factors for the formation of nanoparticles:concen‐tration of chitosan and TPP ,pH value ,N/P ratio were studied by single‐factor experiment ,with nanoparticles size and zeta potential as index .Cell transfection test was carried out to indicate that enhancement of cell transfection efficiency of nano‐car‐rier .Results Nanoparticles loaded DNA vaccine were nearly spherical shape with uniform particle size chitosan nanoparticle (CS) ,(219 .2 ± 18 .2) nm ;quaternary ammonium chitosan nanoparticles(CSTM) ,(222 .5 ± 15 .6) nm .Zeta potential of CS and CSTM was (24 .7 ± 3 .5) mV ,(19 .6 ± 1 .2) mV and encapsulation efficiency was 91 .24% ,87 .66% ,respectively .CSTM nano‐particle could enhance cellular uptake of pVAX1‐wapA obviously .Conclusion CSTM nanoparticle was proved to be an efficient DNA vaccine delivery vector .
ABSTRACT
We designed two novel polymer materials N-glycyrrhetinic acid-polyethylene glycol-chitosan derivatives (NGPC) and N-quaternary ammonium-chitosan derivatives (NQC). We prepared three kinds of drug loaded chitosan nanoparticles (brucine/NGPC-NPs, brucine/NQC-NPs, brucine/MNPs) by ionic crosslinking method with brucine as a model drug and chitosan nanoparticles (brucine/NGPC-NPs, brucine/NQC-NPs) as the reference formulation. Using high content analysis, flow cytometry, immunofluorescence, transmission electron microscopy and other advanced technology, we tested the effect of 20 μg·mL-1 concentration of brucine solution and brucine/chitosan nanoparticles (brucine/CTS-NPs) in hepatocarcinoma (HepG2) cells and evaluated the apoptosis induced by the treatment. The results suggested that brucine-CTS/NPs had a strongest activity in killing tumor cells, and increased the total cell apoptosis rate with a significant formation of "crescent-shaped" body, swelling mitochondria, mitochondria cristae missing, decreased mitochondrial membrane potential and release of cytochrome C. The activity was enhanced by multifunctional nanocomposite particles that increased the cumulative amount of drug in the mitochondria for the anti-tumor effect.
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The current study was undertaken in order to evaluate the efficacy of BSA and chitosan nanoparticle for sustainable delivery of Quercetin. BSA and chitosan nanoparticles were prepared by coacervation process and Ionic gelation method respectively and were characterized by Scanning Electron Microscope. The average size of BSA and chitosan nanoparticles was found to be 96nm and 42.7nm respectively. Quercetin loaded BSA and chitosan nanoparticles were smooth and spherical with the average size of 317.1 nm and 44.1nm respectively. The encapsulation efficiency of the BSA and chitosan nanoparticles were calculated and it was found that the drug encapsulation efficiency was more in chitosan than in BSA. The in vitro analysis of Quercetin loaded BSA and chitosan nanoparticles were studied using dialysis bag. The nanoformulation of Quercetin loaded BSA nanoparticles shows a sustainable release of drug in which 56% of the drug was released till 24 hours and it did not change further. For Quercetin loaded chitosan nanoparticles it shows 72% of in vitro drug release for 24 hours and found to be better than BSA nanoparticle. Hence both Quercetin loaded BSA and chitosan nanoparticles were effective in encapsulation and sustainable release which also paves way for the bio availability of Quercetin
ABSTRACT
Background Posterior capsular opacification (PCO) following cataract extracapsular extraction is a major cause of the reduction of visual acuity.Topical administration of eye drops is a research hotspot for the prevention of PCO.Objective This study was to evaluate the release of cyclosporine A-loaded chitosan nanoparticles (CyA-CS-NP) by ionic gelation in vitro and its feasibility of modification of the surface of polymethylmethacrylate intraocular lens (PMMA IOL) with CyA-CS-NP.Methods The CS-NP and CyA-CS-NP were prepared by ionic gelation of CS with sodium tripolyphosphate.The characteristics of CS-NP,such as the appearance and mean size,and drug entrapment efficient (EE),loading capacity (LC),and the drug release were studied ; the CyA content on the IOL surface was detected by high performance liquid chromatography (HPLC).The IOL surface modified with CyA-CS-NP was observed by scanning electron microscope and X-ray photoelectron spectroscopy technique (XPS),the changes of elements and chemical bond types between simple plasma processed IOL and CyA-CS-NP modified IOL were analyzed.The transmittance at the wavelength of 360-490 nm and refraction of IOL were detected using back focal length method and spectrophotometer,and the effective resolution of IOL was evaluated according to the instruction of ISO/CD 11979-2.Loops anti fatigue test of IOL referred to the criteria of ISO/CD 11979-3.Results The CS-NP and CyA-CS-NP showed monodisperse,uniform appearance similar to spherical shape with a mean size of (158±18) nm and (219±29) nm,respectively.The CyA-CS-NP had high CyA association efficiency and loading capacity (64.2％ and 7.6％).In vitro release study revealed a fast release on the first day followed by a increased drug release during an 11-day following up.The sustained release was approximately 46.6％ at day 1 and 77.7％ at day 12,respectively.The surface of IOLs with cling film was smooth without CS-NP;while the edge of IOLs appeared a layer of CyA-CS-NP after modification.XPS analysis displayed some elements such as phosphonium,CNH2 and O =CN that appeared on the modified surface,indicating that CyA-CS-NP existed on the surface of IOLs edge.The mean quality of CyA on three IOLs surface after modification was 171.88 μg.The diopter,distinguishing ability and transmittance of modified IOL were (16.64±0.23) D,(90.28 ± 0.25) ％ and (73.57 ±0.62) ％,and those of unmodified IOL were (16.62±0.29) D,(90.28±0.21) ％,(73.61±0.60)％,without significant differences between them (t =0.381,0.078,2.291,all at P ＞ 0.05).The antifatigue ability of loops complied with the criteria of ISO/CD 11979-3.Conclusions The optical property and antifatigue ability of loops of the edge-modified IOLs by CyA-CS-NP reach the normal standard and meet the requirement of clinic application.The edge-modified IOLs by CyA-CS-NP can be a delivery system for intraocular drug release.
ABSTRACT
Aim: To prepare chitosan nanoparticlescarrying gene and study its characteristics in vitro and in vivo.Methods:12-ACSs was dissolved in 0.05mol/L sodium acetate buffer to form a solution of 1mg/ml and a DNA(plasmid-encoding antisense ECE ) solution of 0.1mg/ml was dissolved in 25mmol/L Na2SO4.The charge ratio of components is selected as 2/1 for 12-ACSs /DNA complex.The complex was prepared by mixing 12-ACSs solution with DNA solution prior to observation by using transmission electron microscopy.Using Electrophoretic Retardation of DNA Migration detection,DNA precipitation,Binding Equilibration and DNase Resistivity Assay,the formation of 12-ACSs /DNA complex was determined and its stability was simultaneously evaluated.MTT Cell Proliferation Assays was performed on investigation of the cytotoxicity of 12-ACSs /DNA nanoparticles in bronchial epithelial cells (16HBE).The transfection efficiency of 12-ACSs /DNA nanoparticles in vitro and in vivo was investigated in 16HBE cells and Balb/c mice.Results: 12-ACSs /DNA nanoparticles (100~150nm) were observed by transmission electron microscopy.12-ACSs can protect the plasmid encoding antisence-ECE from degradation by DNase I.12-ACSs can transfer plasmid-encoding antisense ECE into 16HBE cells and into the airway epithelium in mice.As shown by fluorescent microscopy,green fluorescent protein reporter can be observed in the transfected cells as well as in the airway epithelium of the treated mice.And it showed a lower cell cytotocixity in cultured 16HBE cells and in mice treated with12-ACSs /DNA nanoparticles.Conclusion: In summery,the chitosan can be used as an effective protectant for DNA as well as an enhancer for in vitro gene transfection.