Distribution of Melaleuca alternifolia essential oil in liposomes with Tween 80 addition and enhancement of in vitro antimicrobial effect

Tea tree oil (TTO) exhibits excellent broad-spectrum antimicrobial activity. In order to preserve it from the degradation in the presence of oxygen, light and temperature, TTO was encapsulated in liposomes (LTTOs) using the thin-membrane hydration and sonication method, and characterised by Zetasizer for size and size distribution, transmission electron microscope for morphology, zeta-potential for surface charge, entrapment efficiency and TTO release from the nanoparticles. The antimicrobial activities of phosphate-buffered saline solution containing TTO, unloaded liposomes and LTTOs suspension were determined by twofold serial broth dilution technique. The size of LTTOs was 75 nm and the encapsulation efficiency of 96.08% was obtained. LTTOs exhibited slow release of TTO and superior broad-spectrum antimicrobial effects compared with free TTO. Liposomes not only effectively encapsulated TTO to form a stable liposome suspension, but also enhanced inhibition and bactericidal effect on the TTO-tolerant strain. The liposomal systems carrying TTO may be a potential alternative for effective antimicrobial agents.     

Formulation of chitosan-TPP-pDNA nanocapsules for gene therapy applications

The encapsulation of DNA inside nanoparticles meant for gene delivery applications is a challenging process where several parameters need to be modulated in order to design nanocapsules with specific tailored characteristics. The purpose of this study was to investigate and improve the formulation parameters of plasmid DNA (pDNA) loaded in chitosan nanocapsules using tripolyphosphate (TPP) as polyanionic crosslinker. Nanocapsule morphology and encapsulation efficiency were analyzed as a function of chitosan degree of deacetylation and chitosan-TPP ratio. The manipulation of these parameters influenced not only the particle size but also the encapsulation and release of pDNA. Consequently the transfection efficiency of the nanoparticulated systems was also enhanced with the optimization of the particle characteristics. Overall, the differently formulated nanoparticulated systems possess singular properties that can be employed according to the desired gene delivery application.     

Novel drug delivery system by surface modified magnetic nanoparticles

In the recent progress of gene and cell therapy, novel drug delivery system (DDS) has been required for efficient delivery of small molecules/drugs and also the safety for clinical usage. We have already developed the unique transfection technique by preparing magnetic vector and using permanent magnet. This technique can improve the transfection efficiency. In this study, we directly associated plasmid DNA with magnetic nanoparticles, which can potentially enhance their transfection efficiency by magnetic force. Magnetic nanoparticle, such as magnetite, its average size of 18.7 nm, can be navigated by magnetic force and is basically consisted with oxidized Fe that is commonly used as the supplement drug for anemia. The magnetite particles coated with protamine sulfate, which gives a cationic surface charge onto the magnetite particle, significantly enhanced the transfection efficiency in vitro cell culture system. The magnetite particles coated with protamine sulfate also easily associated with cell surface, leading to high magnetic seeding percentage. From these results, it was found that the size and surface chemistry of magnetic particles would be tailored to meet specific demands on physical and biological characteristics accordingly. Overall, magnetic nanoparticles with different surface modification enhance the association with plasmid DNA and cell surface as well as HVJ-E, which potentially help to improve the drug delivery system.     

Preparation of PLGA nanoparticles using TPGS in the spontaneous emulsification solvent diffusion method

D-alpha-tocopheryl poly (ethylene glycol) 1000 succinate (TPGS) is a widely used form of vitamin E that has been used as a solubilizer, an emulsifier and as a vehicle for drug delivery formulations. In this study, poly lactide-co-glycolide (PLGA) nanoparticles were prepared by spontaneous emulsification solvent diffusion (SESD) method. TPGS as an emulsifier and further as a matrix material blended with PLGA was used to enhance the encapsulation efficiency and improve the drug release profile of nanoparticles. Rifampicin and estradiol valerate were used as model drugs with different water solubility. The effect of formulation parameters such as drug/polymer ratio, oil phase combination, volume and surfactant content was evaluated. The surface morphology and size of the nanoparticles were studied by scanning electron microscopy (SEM) and laser light scattering. Drug encapsulation efficiency and in vitro drug release profiles of nanoparticles were determined using high performance liquid chromatography (HPLC). The nanoparticles prepared in this study were spherical with size range of 150–250?nm. It was shown that TPGS was a good emulsifier for producing nanoparticles of hydrophobic drugs and improving the encapsulation efficiency and drug loading and drug release profile of nanoparticles. However, the drug loading efficiency of rifampicin, a slightly water-soluble molecule, was significantly lower than that of estradiol valerate, a water insoluble molecule.     

Preparation of dexamethasone-based cationic liposome and its application to gene delivery in vitro

In this study, dexamethasone was conjugated to PAMAM dendrimer (generation 0) and its gene transfection efficiency was investigated. To make a liposomal solution for gene delivery, DOPE was used as a fusogenic helper lipid. In gel retardation assay, PAMAM-dexamethasone conjugate (PAM-Dex)/DOPE liposome/DNA complex was completely retarded at 8:1 N/P (nitrogen/phosphate) ratio. The physicochemical characteristics are studied by measuring the average size distribution and zeta-potential values of the complexes. In vitro transfection assay showed that the PAM-Dex/DOPE liposome/DNA complex displayed higher gene delivery efficiency compared to PAMAM/DNA complex. In addition, PAM-Dex/DOPE liposome showed the lowest toxicity compared to PAMAM, PEI 25 kD and Lipofectamine. These results indicate that PAM-Dex/DOPE liposome has a potential to be used as an efficient gene carrier for gene therapy.     

In vitro antifungal activity and toxicity of itraconazole in DMSA-PLGA nanoparticles

Itraconazole (ITZ) is a drug used to treat various fungal infections and may cause side effects. The aim of this study was to develop and evaluate the in vitro activity of DMSA-PLGA nanoparticles loaded with ITZ against Paracoccidioides brasiliensis, as well as their cytotoxicity. Nanoparticles were prepared using the emulsification-evaporation technique and characterized by their encapsulation efficiency, morphology (TEM), size (Nanosight) and charge (zeta potential). Antifungal efficacy in P. brasiliensis was determined by minimal inhibition concentration (MIC), and cytotoxicity using MTT assay. ITZ was effectively incorporated in the PLGA-DMSA nanoparticles with a loading efficiency of 72.8 +/- 3.50%. The shape was round with a solid polymeric structure, and a size distribution of 174 +/- 86 nm (Average +/- SD). The particles were negatively charged. ITZ-NANO presented antifungal inhibition (MIC = 6.25 ug/mL) against P. brasiliensis and showed lower in vitro cytotoxicity than free drug (ITZ).     

Synthesis of water soluble quantum dots for monitoring carrier-DNA nanoparticles in plant cells

Fluorescent quantum dots (QDs) have shown great promise for use as biolabels in cell and animal biology and more recently in plant sciences. An important use of QDs is for monitoring the dynamics, intracellular trafficking, and fate of carrier-DNA nanocomplexes in cell transfection and potentially in plant transformation. In this study, a low cost aqueous procedure has been developed to efficiently prepare biocompatible QDs for monitoring nanoparticle-mediated gene transfer in conjunction with molecular breeding of Jatropha curcas. Water-soluble CdSe nanoparticles were synthesized by self-assembly using L-Cysteine as stabilizer and optimal synthesis scheme established by fluorescence spectroscopy. The QDs were used to label chitosan-DNA nanoparticles via electrostatic interaction and the resultant QD-labeled chitosan-DNA complexes were shown to have superior fluorescence properties with red shift of emission and absorption spectra relative to the CdSe QDs alone. This system is being explored as a superior alternative to Agrobacterium-mediated genetic transformation of Jatropha curcas cells. PCR amplification of the full length of the carried reporter gene (GFP) suggests that the DNA was not digested in Jatropha curcas cells transfected with CdSe/CS-DNA complexes. Furthermore, GFP gene expression in the transfected callus cells, as evidenced by fluorescence detection, suggests that the target DNA was integrated into the plant genome.     

The influence of physicochemical parameters on the efficacy of non-viral DNA transfection complexes: a comparative study

Various polycationic vehicles have been developed to facilitate the transfer of foreign DNA into mammalian cells. Structure-activity studies suggested that biophysical properties, such as size, charge, and morphology of the resulting DNA complexes determine transfection efficiency within one class of vector. To investigate the general validity of these criteria, we studied the efficacy of a variety of DNA delivery vehicles including liposomes (DOTAP, SAINT2) with and without helper lipid (DOPE), the polymer polyethyleneimine (PEI), and cationic nanoparticles (Si26H, PLGA/chitosan) in a comparative manner. Sizes of the DNA complexes varied between 100 and 500 nm for PEI polyplexes and DOTAP/DOPE lipoplexes, respectively. The zeta potential was positive for PEI, Si26H, and DOTAP based complexes, while it was neutral for SAINT2-DNA complexes and negative for PLGA/chitosan-DNA complexes. The latter finding was elucidated by AFM, showing a layer of DNA adsorbed onto the nanoparticles. Transfection activity was negligible for PLGA/chitosan nanospheres, moderate for Si26H nanospheres and high for all other complexes, PEI being the most active carrier. The liposomal preparations were of low (DOTAP) or moderate (SAINT2) stability in serum, resulting in a pronounced reduction of gene expression, which was partially restored by the addition of chloroquine. In conclusion, transfection efficiency (i) seems to require a positive or neutral zeta potential, (ii) is depending on size, e.g., is higher for smaller particles, and (iii) requires a vector that is stable in serum.     

Encapsulation of hydrophilic and lipophilic drugs in PLGA nanoparticles by the nanoprecipitation method

The purpose of this study was to assess the relative advantages and drawbacks of the nanoprecipitation-solvent displacement method for a range of drugs with respect to the particle size and drug encapsulation in polylactic-co-glycolic acid (PLGA) nanoparticles. The particle size analysis indicated a unimodal particle size distribution in all systems, with a mean diameter of 160-170 nm, except for insulin nanoparticles, which showed a smaller particle size. The results of the encapsulation efficiency analysis demonstrated that more lipophilic drugs, such as cyclosporin and indomethacin, do not suffer from the problems of drug leakage to the external medium, resulting in improved drug content in the nanoparticles. In spite of the fact that valproic acid is a liquid that is very sparingly soluble in water, very low encapsulation efficiency was obtained. Ketoprofen, a drug sparingly soluble in water, demonstrated intermediate values of encapsulation that were well correlated with its intermediate lipophilicity. More hydrophilic drugs, such as vancomycin and phenobarbital, were poorly encapsulated in PLGA nanoparticles. Insulin was preferentially surface bound on the PLGA nanoparticles. However, a strong hypoglycemic effect of the insulin was observed after administration of the suspension of PLGA nanoparticles with surface-bound insulin to the ileum loop of male Wistar rats.     

单分散γ-Al_2O_3纳米颗粒制备工艺参数的调控

采用均相沉淀法,在950℃煅烧碱式硫酸铝(Al(SO_4)_x(OH)_y)纳米颗粒2h,制备出单分散的γ-Al_2O_3纳米颗粒。研究了阴离子、浓度和沉淀剂/铝盐摩尔比对Al(SO_4)_x(OH)_y颗粒的形貌、尺寸和分散性的影响。结果表明,Al(SO_4)_x(OH)_y颗粒的最佳制备条件为:二甲胺硼烷(DMAB)作沉淀剂、Al_2(SO_4)_3.18H_2O作铝源、[Al~(3+)]=0.0005mol/L、n(DMAB)/n(Al_2(SO_4)_3·18H_2O)=40。制备出的Al(SO_4)_x(OH)_y具有球形、平均颗粒尺寸为98nm、粒径分布为75~130nm、单分散性的特点;获得了球形、平均颗粒尺寸为90nm、粒径分布范围为50~105nm、单分散的γ-Al_2O_3颗粒。     

去甲斑蝥素PLA-PEG纳米微球的制备研究

采用复乳法和相分离法两种方法制备去甲斑蝥素的聚乳酸-聚乙二醇嵌段共聚物(PLA-PEG)纳米微球.对比了两种不同方法对制得的含药微球在粒径、包封率以及缓释性能方面的差异.用激光粒度分析仪表征了微球的粒径及其分布,并用透射电镜观察了微球的形貌,其结果表明:复乳法与相分离法制备的微球粒径均在100nm左右,并且成球性好;相对于复乳法,相分离法制备的微球分布较宽,包封率较高,可达到50%左右;体外释放实验表明两种方法制备的微球都具有缓释作用.     

Preparation of embolic NEMs loading capecitabine

The nanoparticles-embedded microcapsules (NEMs) with smooth surface, good sphericity, excellent dispersivity and uniform particle size distribution were prepared by emulsification combined with electrospraying to extend the sustained release performance of the embolic microcapsules loading capecitabine (CAP). The sodium alginate and chitosan with good biocompatibility were used as the materials and CAP as a small-molecule model drug. The drug loading, encapsulation efficiency and drug release of CAP in the NEMs were investigated. The results showed that the drug-loading and encapsulation efficiency both increased with the increment of chitosan and CAP concentration. The maximum values of drug loading and encapsulation efficiency were 1.97 and 18.01 % respectively when initial CAP concentration was 5.0 g/L and chitosan molecular weight 100 kDa. The cumulative release rate of CAP released from the NEMs was lower than 30 % in 0.5 h, which indicated that there was no obvious initial burst release behavior. In the subsequent 240 h, the release results confirmed that the NEMs had better sustained release properties compared to pure microcapsules, and it might be a new anticancer drug delivery system in the future studies.     

Novel gelatin-siloxane nanoparticles decorated by Tat peptide as vectors for gene therapy

In principle, the technique of gene delivery involves taking complete or parts of genes that can code specific messages and delivering them to selected cells in the body. Such a transfer of plasmid DNA into mammalian cells has posed major challenges for gene therapy. A series of gelatin-siloxane nanoparticles (GS NPs) with controlled size and surface charge were synthesized through a two-step sol-gel process. In order to increase the efficiency of cellular uptake, HIV-derived Tat peptide was further grafted to GS NPs. In vitro co-location and endocytosis inhibition experiments suggested that the as-synthesized TG NPs may enter HeLa cells via a combined pathway of lipid-raft-?and receptor-dependent endocytosis, and only cause little cell damage. Moreover, this study shows the encapsulation of a plasmid DNA in TG NPs to be obtained as a non-viral gene vector. This kind of encapsulation provides complete protection to the plasmid DNA from the external DNase and serum environment, and generates the hope that the resulting formulation can be developed into a potential vector for effective gene delivery. In order to check this potential, the reporter gene pSVβ-gal was encapsulated, and in vitro transfection efficiency of this system was found to be nearly 130% compared to the commercially available transfection reagent Lipofectamine?.     

玉米醇溶蛋白–香芹酚纳米颗粒的制备及其性能研究

目的 利用天然抗菌剂来保护食品品质和延长食品的货架期。方法 研究利用玉米醇溶蛋白自组装的特性包覆香芹酚植物精油,然后用酪蛋白酸钠作为稳定剂来制备载香芹酚复合纳米粒子。结果 玉米醇溶蛋白纳米颗粒对香芹酚具有良好的包封率(71.52%～80.09%)。扫描电子显微镜(SEM)显示香芹酚复合纳米粒子分布均匀,呈球形,粒径为80～220 nm。同时,包覆香芹酚的纳米粒子具有良好的复溶性、储存稳定性和抗氧化性能,并且对金黄色葡萄球菌、大肠杆菌表现出良好的抗菌性能。结论 将该复合纳米颗粒应用于食品包装中,可有效地提高食品的抗氧化特性和抗菌性,抑制食品腐败变质、延长食品货架期,在食品工业中具有潜在的应用前景。     

Thymopentin-loaded pH-sensitive chitosan nanoparticles for oral administration: preparation, characterization, and pharmacodynamics

Thymopentin, a potent immunomodulating drug, was incorporated into pH-sensitive chitosan nanoparticles prepared by ionic gelation of chitosan with tripolyphosphate anions and then coated with Eudragit S100 to improve the stability and the oral bioavailability. Nanoparticles particle size and zeta potential were measured by photo correction spectroscopy and laser Dopper anemometry. Its morphology was examined by environment scan electron microscope. The encapsulation efficiency and the release in vitro were determined by HPLC. Enzymatic stabilization was expressed by the enzymatic degradation of aminopeptidase. Biological activity of TP5 loaded in nanoparticles was assayed by lymphocyte proliferation test in vitro and the immune function (CD4+/CD8+) of irradiated rat in vivo. The results obtained demonstrated that the average sizes of pH-sensitive chitosan nanoparticles were 175.6 +/- 17 nm, the zeta potential was 28.44 +/- 0.5 mV and the encapsulation efficiency was 76.70 +/- 2.6%. The cumulative release percentages of thymopentin from the pH-sensitive nanoparticles were 24.65%, 41.01%, and 81.44% incubated in different medium, 0.1 N HCl, pH 5.0 PBS, and pH 7.4 PBS, respectively. The pH-sensitive chitosan nanoparticles could efficiently protect TP5 from enzymatic degradation and prolong the degradation half-time of TP5 from 1.5 min to 15 min. It was demonstrated from the lymphocyte proliferation test that the nanoparticle-encapsulated TP5 still kept its biological activity. In immunosuppression rats, the lowered T-lymphocyte subsets values were significantly increased and the raised CD4+/CD8+ ratio was evidently reduced. These results indicated that pH-sensitive chitosan nanoparticles may be used as the vector in oral drug delivery system for TP5.     

Controlled release of insulin from PLGA nanoparticles embedded within PVA hydrogels

A simple and versatile delivery platform for peptide and protein based on physically cross-linked poly (vinyl alcohol) (PVA) hydrogels containing insulin-loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles was successfully fabricated. The particle morphology and size were characterized by SEM and laser light scattering method, respectively. Results showed that these particles had a mean diameter of 615 nm with a narrow size distribution and homogeneous particle production. The protein encapsulation efficiency was 72.6%. When insulin-loaded PLGA nanoparticles were administered intraperitoneally as a single dose (20 U/kg) to streptozotocin-induced diabetic mouse, blood glucose levels of these mice decreased and it could be sustained at such levels over 24 h. In vitro release further indicated that entrapment of the nanoparticles into the PVA hydrogels causes a reduction in both the release rate and the total amount of insulin released, which suggesting that PLGA nanoparticles entrapped into the PVA hydrogels showed more suitable controlled release kinetics for protein delivery.     

Continuous Production of Discrete Plasmid DNA‐Polycation Nanoparticles Using Flash Nanocomplexation

Despite successful demonstration of linear polyethyleneimine (lPEI) as an effective carrier for a wide range of gene medicine, including DNA plasmids, small interfering RNAs, mRNAs, etc., and continuous improvement of the physical properties and biological performance of the polyelectrolyte complex nanoparticles prepared from lPEI and nucleic acids, there still exist major challenges to produce these nanocomplexes in a scalable manner, particularly for lPEI/DNA nanoparticles. This has significantly hindered the progress toward clinical translation of these nanoparticle‐based gene medicine. Here the authors report a flash nanocomplexation (FNC) method that achieves continuous production of lPEI/plasmid DNA nanoparticles with narrow size distribution using a confined impinging jet device. The method involves the complex coacervation of negatively charged DNA plasmid and positive charged lPEI under rapid, highly dynamic, and homogeneous mixing conditions, producing polyelectrolyte complex nanoparticles with narrow distribution of particle size and shape. The average number of plasmid DNA packaged per nanoparticles and its distribution are similar between the FNC method and the small‐scale batch mixing method. In addition, the nanoparticles prepared by these two methods exhibit similar cell transfection efficiency. These results confirm that FNC is an effective and scalable method that can produce well‐controlled lPEI/plasmid DNA nanoparticles.     

Synthesis of multilayer Nano-ZrO2 coated polystyrene spheres on fabrication of three-dimensional ordered macroporous structures

Monodispersed ZrO₂ precursor nanoparticles with a diameter of 25 nm were successfully synthesized by using diglycol as complexing reagent. The kinetic of particle growth as a function of concentration ratio of ZrOCl₂: diglycol was investigated. The as-synthesized ZrO₂ precursor nanoparticles were homogenously coated on the surface of polystyrene particles. Multilayer coating process was implemented by using poly (acrylic acid) (PAA) to modify the surface charges of the coated spheres, which was characterized by zeta-potential, particles size distribution and microstructural observation. The multilayer-coated polystyrene (PS) spheres have been used as templates to produce macroporous materials. Ordered macroporous ZrO₂ materials were obtained after the ZrO₂ precursor nanoparticles coated PS spheres were formed by centrifugation and calcined at 550°C for 3 h. The porous wall thickness could be well controlled by using the multilayer nano-ZrO₂ coated PS spheres with different coating thickness.     

Biocompatible nanoclusters of <Emphasis Type="Italic">O</Emphasis>-carboxymethyl chitosan-coated Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> nanoparticles: synthesis,characterization and magnetic heating efficiency

In this work, we developed a polymer encapsulation of Fe₃O₄ nanoparticles as a core–shell nanocluster with different sizes to investigate the cluster structure effect on their magnetic properties and magnetic heating behavior. Well-dispersed nanoclusters of O-carboxymethyl chitosan-coated Fe₃O₄ nanoparticles were synthesized by microwave-assisted co-precipitation. The cluster sizes were tunable by varying the concentration of polymers used during synthesis. Nanoclusters present superparamagnetic behavior at room temperature with a reduction in saturation magnetization as a consequence of coating layer. The shift of blocking temperature to the higher value with increasing clusters size shows the stronger magnetic interaction in larger magnetic clusters. In a low alternating magnetic field with frequency of 178 Hz and amplitude of 103 Oe, nanoclusters offer a high heating efficiency. A maximum specific absorption rate of 204 W/g is observed in the sample with hydrodynamic size of 53 nm. In vitro cytotoxicity analysis performed on HeLa cells verified that nanoclusters show a good biocompatibility and can be an excellent candidate for applications in hyperthermia cancer treatment.     

Conjugation of membrane-destabilizing peptide onto gelatin–siloxane nanoparticles for efficient gene expression

One of the crucial steps in gene delivery with non-viral vectors is the escape of DNA complexes from the endosome. In order to improve gene transfection efficiency, we designed a novel gene delivery vector gelatin–siloxane nanoparticles (GS NPs) conjugated with two different membrane-destabilizing peptides, octaarginine (R8) and a subunit of influenza virus haemagglutinin HA2. Both R8-GS NPs and HA2-GS NPs had high positive surface charges. They could condense and protect DNA against serum/DNase degradation. Results from flow cytometry and confocal laser scanning microscope respectively indicated that R8-GS NPs had higher uptake efficiency than HA2-GS NPs, whereas HA2-GS had higher endosome escaping efficiency. Furthermore, in vitro transfection displayed a higher gene expression level with HA2-modified GS NPs, which suggested that endosome escaping is the crucial step for nanoparticle mediated gene therapy.