On-demand preparation of quantum dot-encoded microparticles using a droplet microfluidic system

Optical barcoding technology based on quantum dot (QD)-encoded microparticles has attracted increasing attention in high-throughput multiplexed biological assays, which is realized by embedding different-sized QDs into polymeric matrixes at precisely controlled ratios. Considering the advantage of droplet-based microfluidics, producing monodisperse particles with precise control over the size, shape and composition, we present a proof-of-concept approach for on-demand preparation of QD-encoded microparticles based on this versatile new strategy. Combining a flow-focusing microchannel with a double T-junction in a microfluidic chip, biocompatible QD-doped microparticles were constructed by shearing sodium alginate solution into microdroplets and on-chip gelating these droplets into a hydrogel matrix to encapsulate CdSe/ZnS QDs. Size-controllable QD-doped hydrogel microparticles were produced under the optimum flow conditions, and their fluorescent properties were investigated. A novel multiplex optical encoding strategy was realized by loading different sized QDs into a single droplet (and thus a hydrogel microparticle) with different concentrations, which was triggered by tuning the flow rates of the sodium alginate solutions entrapped with different-colored QDs. A series of QD-encoded microparticles were controllably, and continuously, produced in a single step with the present approach. Their application in a model immunoassay demonstrated the potential practicability of QD-encoded hydrogel microparticles in multiplexed biomolecular detection. This simple and robust strategy should be further improved and practically used in making barcode microparticles with various polymer matrixes.     

Characterization and biological evaluation of paclitaxel-loaded poly(L-lactic acid) microparticles prepared by supercritical CO2

In this study, paclitaxel loaded poly( L-lactic acid) (PTX-PLLA) microparticles were prepared using solution enhanced dispersion by supercritical CO2(SEDS) technique. This supercritical antisolvent technique offers the advantage of negligible organic solvent residua in the drug loaded microparticles. Scanning electron microscopy (SEM) showed that microparticles exhibited rather spherical shape and small particle size with narrow particle size distribution. X-ray diffraction (XRD) and differential scanning calorimeter (DSC) indicated that PTX was amorphously dispersed in the PLLA matrix. The drug loading and encapsulation efficiency of PTX-PLLA microparticles were 14.33% and 62.68%, respectively. In vitro cytotoxicity evaluation of PTX-PLLA microparticles against nonsmall-cell lung cancer A549 and ovarian cancer SKOV3 cell lines indicated that PTX-PLLA had superior antiproliferation activity against the A549 and SKOV3 cell lines, compared with free PTX formulations. The cellular internalization of fluorescent microparticles was evidenced by fluorescence microscope and further confirmed by transmission electron microscopy (TEM). This was attributed to the efficient intracellular accumulation of PTX via cell phagocytosis and sustained release of PTX from PLLA matrix. The anticancer activity of PTX-PLLA was associated with PTX-induced cell apoptosis such as nuclear aberrations, condensation of chromatin and swelling damage in mitochondria. The cell apoptosis index detected by flow cytometry was higher in PTX-PLLA group than in free PTX. The PTX-PLLA formulation, which was obtained through micronization of PTX and encapsulation of micronized PTX into PLLA simultaneously in the SEDS process, significantly potentiated the anticancer activity of PTX.     

Comparison of molecular imprinted particles prepared using precipitation polymerization in water and chloroform for fluorescent detection of nitroaromatics

A comparative study was conducted to study the effects that two different polymerization solvents would have on the properties of imprinted polymer microparticles prepared using precipitation polymerization. Microparticles prepared in chloroform, which previous results indicated was the optimal solvent for molecular imprinting of nitroaromatic explosive compounds, were compared to water, which was hypothesized to decrease water swelling of the polymer and allow enhanced rebinding of aqueous template. The microparticles were characterized and were integrated into a fluorescence sensing mechanism for detection of nitroaromatic explosive compounds. The performance of the sensing mechanisms was compared to illustrate which polymerization solvent produced optimal imprinted polymer microparticles for detection of nitroaromatic molecules. Results indicated that the structures of microparticles synthesized in chloroform versus water varied greatly. Sensor performance studies showed that the microparticles prepared in chloroform had greater imprinting efficiency and higher template rebinding than those prepared in water. For detection of 2,4,6-trinitrotoluene, the chloroform-based fluorescent microparticles achieved a lower limit of detection of 0.1 μM, as compared to 100 μM for the water-based fluorescent microparticles. Detection limits for 2,4-dinitrotoluene, as well as time response studies, also demonstrated that the chloroform-based particles are more effective for detection of nitroaromatic compounds than water-based particles. These results illustrate that the enhanced chemical properties of using the experimentally determined optimal polymerization solvent overcome deformation of imprinted binding sites by water swelling and benefits of using the polymerization solvent for rebinding of the template.     

SiO2/聚甲基丙烯酸叔丁酯核壳复合微粒的制备与表征

以甲基丙烯酸-3-(三甲氧基硅基)丙酯(MPS)修饰的SiO2胶体粒子为种子,甲基丙烯酸叔丁酯(tBMA)为单体、十二烷基硫酸钠(SDS)为乳化剂,采用种子乳液聚合法制备了SiO2/聚甲基丙烯酸叔丁酯的核壳复合微粒。微粒经水解后形成具有pH敏感性的无机/有机复合微粒。研究了影响核壳复合微粒形态结构的因素,结果发现,控制SiO2种子乳液的质量分数在1.5%～2%,可避免聚合过程中生成纯聚甲基丙烯酸叔丁酯乳胶粒子;反应体系中乳化剂SDS的用量超过质量分数0.3%时,易形成纯聚合物乳胶粒子;SDS用量低于质量分数0.15%时,生成的核壳复合微粒易产生团聚;单体和交联剂用量升高,核壳复合微粒的壳层厚度增加,用量过高会导致核壳复合微粒出现团聚现象,并且有纯聚合物乳胶粒子生成。采用TEM、NMR和FTIR及接触角测试技术分析结果表明,复合微粒是由SiO和聚甲基丙烯酸叔丁酯组成的核壳结构微粒。     

Novel separation and preconcentration of trace amounts of copper(II) in water samples based on neocuproine modified magnetic microparticles

A novel, simple method based on magnetically assisted chemical separation (MACS) has been developed for analytical purposes. In this method, neocuproine modified magnetic microparticles was used for selective extraction and preconcentration of copper(II) ions from aqueous solutions. The advantages of this method include consumption of organic solvents almost eliminated and applications on unclear (containing suspended particles) samples without any preliminary filtration step. This method combines simplicity and selectivity of solvent extraction with easy separation of magnetic microparticles from solution with magnet. In addition, it can be considered as a simple method for determination of partition coefficient. The influence of different parameters, such as presence of extractant, amount of extractant loaded on the microparticles, reducing agent, pH, equilibrium time, ionic strength, type and least amount of stripping solution and limit of detection, were evaluated. Also, the effects of various cationic and anionic interferences on the percent recovery of copper were studied. Copper ions were extracted from solution at pH 6 and were stripped from microparticles with 0.5 M HNO₃. Extraction efficiencies for solutions with volumes up to 100 ml were >99%. Limit of detection was 1.5 μg/l. The method was applied to the recovery and determination of copper in different water samples.     

A continuous DC-insulator dielectrophoretic sorter of microparticles

A lab-on-a-chip device is described for continuous sorting of fluorescent polystyrene microparticles utilizing direct current insulating dielectrophoresis (DC-iDEP) at lower voltages than previously reported. Particles were sorted by combining electrokinetics and dielectrophoresis in a 250 μm wide PDMS microchannel containing a rectangular insulating obstacle and four outlet channels. The DC-iDEP particle flow behaviors were investigated with 3.18, 6.20 and 10 μm fluorescent polystyrene particles which experience negative DEP forces depending on particle size, DC electric field magnitude and medium conductivity. Due to negative DEP effects, particles are deflected into different outlet streams as they pass the region of high electric field density around the obstacle. Particles suspended in dextrose added phosphate buffer saline (PBS) at conductivities ranging from 0.50 to 8.50 mS/cm at pH 7.0 were compared at 6.85 and 17.1 V/cm. Simulations of electrokinetic and dielectrophoretic forces were conducted with COMSOL Multiphysics^® to predict particle pathlines. Experimental and simulation results show the effect of medium and voltage operating conditions on particle sorting. Further, smaller particles experience smaller iDEP forces and are more susceptible to competing nonlinear electrostatic effects, whereas larger particles experience greater iDEP forces and prefer channels 1 and 2. This work demonstrates that 6.20 and 10 μm particles can be independently sorted into specific outlet streams by tuning medium conductivity even at low operating voltages. This work is an essential step forward in employing DC-iDEP for multiparticle sorting in a continuous flow, multiple outlet lab-on-a-chip device.     

Silicone-modified starch/protein microparticles: protecting biopolymers with a hydrophobic coating

Silicone-coated starch/protein (human serum albumin, HSA) microparticles were prepared by precipitation of a starch/HSA/DMSO/water (water-in-oil) emulsion into acetone containing a silicone: the silicone polymer was either unfunctionalized (SiMe₃ terminated, PDMS) or functionalized at its termini with Si(OEt)₃ groups (TES-PDMS). The microparticles of approximate diameter 2–7 μm were highly hydrophobic with advancing contact angles 115°. Over several minutes, however, the contact angle decreased to ca. 40–70°. Soxhlet extraction with water led to degradation of the microparticles, irrespective of the nature of their silicone coating, as evidenced by release of the protein from them. Intraperitoneal (IP) or gastric administration of the two different particles to mice, however, showed a clear difference between the two silicones. The microparticles coated with either PDMS or TES-PDMS led to very different immune responses. Oral administration of the microparticles prepared with functionalized silicone elicited a significant production of antibodies, whereas the particles prepared with the unfunctionalized silicone (PDMS) were only weakly active. By contrast, the IP results demonstrated that particles coated with PDMS elicited an immune response that was established much more rapidly than with the particles modified with TES-PDMS. It is proposed that the TES-PDMS forms a physically adhering film or covalent bond to the protein molecules, which serves to protect the microparticle from biological degradation in the gut and/or facilitates the microparticle/protein interaction with the immune system.     

Caffeine loading into micro- and nanoparticles of mesoporous silicate materials: in vitro release kinetics

Caffeine was loaded into microparticles and nanoparticles of MCM-41 and MCM-48 to study its release into simulated body fluid at pH 7.4 and 37°C. The silicate systems were characterized by X-Rar Diffraction (XRD), scanning electron microscopy, and nitrogen adsorption/desorption isotherms. Loading of caffeine was confirmed by thermal gravimetric analysis and FTIR and the loading capacity was determined by high performance liquid chromatography (HPLC). The loading capacities for the microparticles are higher than that for the nanoparticles for both silicate systems, with the highest (56.8%) for the microparticles of MCM-48. However, for each system, the release from nanoparticle is faster than from microparticles. For all systems, diffusion is the major dissolution mechanism.     

Screening of mucoadhesive microparticles containing hydroxypropyl-beta-cyclodextrin for the nasal delivery of risperidone

Interaction of the antipsychotic drug risperidone with hydroxypropyl-beta-cyclodextrin (HPBCD) in solution and in the solid state was studied with the aim of overcoming the limitations associated with nasal administration of low solubility drugs. Risperidone solubility studies revealed inclusion complex formation with a 1:1 stoichiometry. Low concentrations (0.1 w/v %) of hydroxypropylmethyl cellulose (HPMC) and carbomer affected risperidone solubility in water. No formation of a ternary complex was detected. The solid inclusion complex was prepared by spray drying and was characterised by thermal (DSC) and spectral (FTIR) analyses. Risperidone and the inclusion complex were loaded into microparticles by spray drying using HPMC, carbomer and HPMC/carbomer interpolymer complex (IPC) as mucoadhesive components. The microparticles were characterised with respect to drug loading, particle size distribution, thermal analysis, and zeta potential measurements. Mucoadhesive properties of the microparticles were studied by measuring the work of adhesion. Carbomer and IPC based microparticles revealed superior mucoadhesive microparticles compared to HPMC based microparticles. Drug incorporation into microparticles reduced their mucoadhesive properties, while incorporation of the cyclodextrin complex caused no additional reduction in mucoadhesion. The in vitro dissolution studies showed that formation of the inclusion complex significantly increased the risperidone dissolution rate from the microparticles, thus providing sustained drug release.     

Composite Hydrogels for Sustained Release of Therapeutic Agents

The goal of this research is to develop a composite hydrogel system for sustained release of therapeutic agents. The hydrogel composites were prepared by embedding drug‐loaded, biodegradable poly (DL‐lactide‐co‐glycolide) (PLGA) microparticles in semicrystalline hydrogels of polyvinyl alcohol (PVA). The gels were physically cross‐linked by the formation of the crystallites. The presence of the crystallites and the composite nature of the structure were confirmed by using differential scanning calorimetry and ATR‐FTIR spectroscopy. The distribution of microparticles in the hydrogel matrix was evaluated by using confocal laser scanning microscopy with coumarin‐6 as a fluorescence marker. The numbers of particles in the hydrogel matrix increased along the scanning depth, indicating uneven distribution. The release behavior of a model therapeutic agent, hydrocortisone, was evaluated, and the hydrogel composite system provided for better control of release than the microparticles and hydrogels alone. The addition of outer layers of PVA to the original single‐layer composite further reduced the initial burst effect from the microparticles and allowed for a linear release profile for greater than 1 month.     

Acetal-derivatized dextran: an acid-responsive biodegradable material for therapeutic applications

Dextran, a biocompatible, water-soluble polysaccharide, was modified at its hydroxyls with acetal moieties such that it became insoluble in water but freely soluble in common organic solvents enabling its use in the facile preparation of acid-sensitive microparticles. These particles degrade in a pH-dependent manner: FITC-dextran was released with a half-life at 37 degrees C of 10 h at pH 5.0 compared to a half-life of approximately 15 days at pH 7.4. Both hydrophobic and hydrophilic cargoes were successfully loaded into these particles using single and double emulsion techniques, respectively. When used in a model vaccine application, particles loaded with the protein ovalbumin (OVA) increased the presentation of OVA-derived peptides to CD8+ T-cells 16-fold relative to OVA alone. Additionally, this dextran derivative was found to be nontoxic in preliminary in vitro cytotoxicity assays. Owing to its ease of preparation, processability, pH-sensitivity, and biocompatibility, this type of modified dextran should find use in numerous drug delivery applications.     

Tailoring of the external and internal morphology of poly-3-hydroxy butyrate microparticles

Some microencapsulation procedures such as oil-in-water (o/w) and water-in-oil-in-water (w/o/w) emulsions were selected in an attempt to produce tailored poly-3-hydroxybutyrate (PHB) microparticles. The effects of several processing parameters such as polymer precipitation, surfactant, solvent, stirring and solvent evaporation rates were also considered. As a rule, low stirring rates at 500 rpm yielded particles ranging between 100 to 250 μm and at rates over 8000 rpm, diameters around 5–10 μm. The surfaces of the bigger particles, observed by scanning electron microscopy (SEM), were rough and the smaller ones were even rougher, irregular, cauliflower like. The extraction of the chloroform under low pressure or to the open atmosphere did not produce any appreciable change in the morphology for either type of particle. Transmission electron microscopy (TEM) micrographs suggest that microparticles obtained by o/w emulsions are monolithics but those obtained by w/o/w emulsions are of capsule-like structure. Microencapsulation of a peptide material such as follicle stimulating hormone was carried out with success using a double emulsion technique. This biomaterial, dissolved in the inner aqueous phase, was able to stabilize the primary emulsion without using a surfactant.     

Layer-by-Layer Construction of Novel Biofunctional Fluorescent Microparticles for Immunoassay Applications

A novel class of biofunctional fluorescent microparticles for application in immunoassays was constructed by using the layer-by-layer self-assembly method to deposit multiple layers of fluorescently labeled polyelectrolytes onto colloidal particles, followed by deposition of a protein (immunoglobulin G, IgG) layer. Microelectrophoresis experiments revealed alternating negative and positive zeta-potentials with deposition of each successive polyelectrolyte layer, indicating that the alternate electrostatic adsorption of polyelectrolytes of opposite charge was successfully achieved. Transmission electron microscopy images showed a change of the particle surface texture after polyelectrolyte multilayer deposition. Fluorescence microscopy image (FMI) analysis provided direct measurement of the fluorescence intensity of single microparticles. The observed systematic increase of the fluorescence intensity of individual microparticles with increasing polyelectrolyte layer number from FMI analysis further demonstrated the controlled regular adsorption of polyelectrolyte layers onto the polystyrene (PS) particles. Protein immobilization onto the polyelectrolyte multilayer-coated particles was verified by the different surface properties of the microparticles with respect to surface charge under pH conditions above and below the isoelectric point of the proteins. The assembly of IgG and fluorescein isothiocyanate-labeled IgG onto polyelectrolyte multilayer-coated PS microparticles and their potential use was ultimately confirmed by a solid phase immunotest. Copyright 2001 Academic Press.     

Hydrothermal Transformations of Ascorbic Acid

Hydrothermal treatment of aqueous solution of ascorbic acid has resulted in the formation of soluble and insoluble products. The soluble product (carbon quantum dots) has exhibited pronounced fluorescent properties. The insoluble product consists of densely aggregated 1–2 μm particles. The increase in the hydrothermal treatment duration has resulted in continuous transformation of carbon quantum dots into the microparticles.     

Doxorubicin-loaded PLGA microparticles with internal pores for long-acting release in pulmonary tumor inhalation treatment

Doxorubicin(DOX) loaded poly(lactic-co-glycolic acid)(PLGA) microparticles with internal pores(MP-D) were developed for long-acting release in pulmonary inhalation treatment. The PLGA microparticles exhibited favorable aerodynamic properties for pulmonary delivery. In vitro drug release profile suggested that MP-D have the advantage of long-term maintenance of drug concentrations. MTT assay demonstrated the in vitro anti-tumor efficiency of the DOX loaded PLGA microparticles. Furthermore, melanoma lung metastasis model was established to determine the in vivo antitumor efficiency. The mice treated with MP-D showed significantly fewer lesions than the untreated ones. The survival analysis indicated that MP-D prolonged the survival time of tumor-bearing mice. These results suggested that DOX loaded PLGA microparticles with internal pores have the potential to be used as long-acting release carriers in clinical lung cancer treatment.     

Formulation optimization and in vitro antibacterial ability investigation of azithromycin loaded FDKP microspheres dry powder inhalation

Azithromycin loaded fumaryl diketopiperazine(FDKP) dry powder inhalation was designed and prepared for the treatment of community-acquired pneumonia.The solubility of FDKP and stability of azithromycin solution was investigated.Formulation of azithromycin loaded FDKP microparticle was investigated and optimized by the single factor experiment.High-pressure homogenization and spray drying conditions were also optimized to prepare the particles by spray drying azithromycin dissolved FDKP microparticle suspension at pH 4.5.The in vitro antibacterial efficiency and in vitro dispersion performance was also investigated to confirm the antibacterial efficiency,dispersion and deposition behavers.FDKP/azithromycin mass ratio(3:2) was the optimized formulation of azithromycin loaded FDKP microparticle with the maximal drug loading efficiency.High-pressure homogenization and spray drying conditions were also optimized.The in vitro antibacterial results indicated that only with the antibiotic concentration higher than mutant prevention concentration could totally inhibit the reproduction of bacteria.In vitro dispersion performance of azithromycin loaded FDKP microparticles(AZM@FDKP-MPs) also shows remarkable improvement of dispersion and deposition behavers of AZM.AZM@FDKP-MPs dry powder inhalation as a targeting delivery route has better potential for lung infection treatment.     

Preparation of microparticles composed of cinnamoyl gelatin and cinnamoyl alginate by spray-drying method and effect of UV irradiation and pH value on their release property

Microparticles composed of cinnamoyl gelatin (CinGel) and cinnamoyl alginate (CinAlg) were prepared as a UV irradiation- and pH-responsive carrier by a spray-drying method. CinGel and CinAlg were prepared by a condensation reaction. Using a colorimetric method, the gelatin to CA mass ratio and the alginate to CA mass ratio were calculated to be 1:0.04 and 1:0.02, respectively. The complexation of CinGel and CinAlg in aqueous solution took place when the medium was acidic (e.g., pH 4.0) and the maximum complexation was obtained when the CinGel/CinAlg mass ratio was 7:3. On SEM photographs, some spray-dried CinGel/CinAlg particles were spherical and others were irregularly shaped, and the diameter was a few to tens of micrometers. The release degrees of amaranth loaded in UV-treated microparticles were lower than those of the dye loaded in UV-untreated ones, possibly because of the photo cross-linkage of CinGel and CinAlg. Regardless of whether the microparticles were UV-treated, the release degree at pH 4.0 was significantly lower than the release degree at pH 6.0 and pH 8.0, possibly because of complexation of CinGel and CinAlg under the acidic condition.     

交联壳聚糖水凝胶填充层状微胶囊的制备与性能

在甲基丙烯酸和乳酸接枝修饰的水溶性壳聚糖(CML)存在下, 合成了尺寸均匀的球形CML杂化碳酸钙微粒. 通过层层组装(LBL)技术在该微粒表面形成了聚苯乙烯磺酸钠(PSS)/聚烯丙基胺盐酸盐(PAH)多层膜, 去除碳酸钙微粒后得到内部含有CML的聚电解质微胶囊. 进一步采用紫外光引发CML聚合, 将CML转化为CML微凝胶, 得到内部填充凝胶的微胶囊. 通过扫描电镜、光学显微镜和透射电镜等技术表征了微胶囊的结构. 与传统的LBL微胶囊不同, 凝胶填充的微胶囊干燥时尺寸收缩, 但仍可保持球形; 再次水化后, 能够膨胀恢复其原有尺寸和形态. 各种具有不同电荷性质、分子量和亲疏水性的染料分子及蛋白质均可有效地装载到微胶囊内.     

Evaluation of retention and release processes of two antibiotics from the biocompatible core-shell microparticles

By dropwise addition of a chitosan solution into different non-solvent, such as: 1 N and 2 N NaOH as well as 1 N NaOH: C₂H₅OH mixture (2:1, v/v) at temperature of 25 °C and 50 °C under stirring, the spherical pure chitosan microparticles were performed. As solvents for chitosan was used 0.1 N acetic acid or 0.1 N HCl. The immersion of the pure chitosan microparticles in hyaluronan solution led to complex microparticles, namely chitosan microparticles covered by a hyaluronan layer. For all the microparticles performed the behaviours in the retention process of two antibiotics: chloramphenicol succinate sodium salt and cefotaxime sodium salt were analyzed. Also, the study shows the release behaviour of cefotaxime sodium salt by the microparticles loaded with this drug. Among the microparticles performed a type of complex microparticles can be considered a suitable drug delivery system for cefotaxime. These microparticles were performed by dropwise addition of chitosan solution in 0.1 N acetic acid into the 1 N NaOH: C₂H₅OH (2:1, v/v) non-solvent at 20 °C for 3 h, followed by their washing up to alkalinity loss and the immersion in hyaluronan solution of 10 g/L concentration for 24 h.     

Synthesis, assembly and reaction of a nanocatalyst in microfluidic systems: a general platform

We present a successive microfluidic approach to create and characterize hierarchical catalyst structures consisting of metal-decorated nanoparticles that are assembled into porous microparticles ("supraball" catalysts). First, using a silicon microreactor, platinum nanoparticles with a very narrow size distribution are grown and immobilized uniformly onto iron oxide/silica core-shell nanospheres. The Pt-decorated silica nanospheres are then assembled into uniform, spherical, micron-sized particles by using emulsion templates generated with a microfluidic drop generator. Finally, the assembled supraballs are loaded into a packed-bed microreactor for characterization of the catalytic reactivity. The prepared material showed excellent catalytic activity for the oxidation of aldehyde with only ～1 mg of material (containing ～50 μg of platinum nanoparticles). After the reactions, all the supraball catalysts are recovered by using the magnetic property of the underlying iron oxide/silica core-shell nanospheres.