Scalable Production of Biomedical Microparticles via High-Throughput Microfluidic Step Emulsification

Drug microcarriers are widely used in disease treatment, and microfluidics is well established in the preparation of microcarrier particles. A proper design of the microfluidic platform toward scalable production of drug microcarriers can extend its application values in wound healing, where large numbers of microcarriers are required. Here, a microfluidic step emulsification method for the preparation of monodisperse droplets is presented. The droplet size depends primarily on the microchannel depth rather than flow rate, making the system robust for high-throughput production of droplets and hydrogel microparticles. Based on this platform, basic fibroblast growth factor (bFGF) is uniformly encapsulated in the microparticles, and black phosphorus (BP) is incorporated for controllable release via near-infrared (NIR) stimulation. The microparticles serve as drug carriers to be applied to the wound site, inducing angiogenesis and collagen deposition, thereby accelerating wound repair. These results indicate that the step emulsification technique provides a promising solution to scalable production of drug microcarriers for wound healing as well as tissue regeneration.     

Innovations in high throughput manufacturing of uniform emulsions and capsules

Membrane emulsification has been widely used in the manufacture of uniform soft and hard spherical particles. It is used to create uniform emulsion droplets whose sizes can be closely controlled. A disperse liquid phase is pressurised to permeate into the pores of a membrane, forming droplets in a drop-by-drop manner in a continuous phase on the other side of the membrane. The droplets formed are detached by applying well controlled detachment forces, which are result from the cross-flow of the continuous phase over the membrane surface, or the rotation of the membrane in the continuous phase. These two technologies are called cross-flow and rotating membrane emulsification, respectively. This paper presents examples of uniform complex spheres, sized from sub-micrometers to a few hundred micrometers, prepared using a pilot scale cross-flow membrane emulsification rig and a bentchtop rotating reactor. Emulsion stabilisation strategies vary from using small molecular surfactants, nanoparticles to surfactant free interfacial polymerisation. The examples demonstrate the advantages and versatility in formulation and manufacture of precisely size- and structure-controlled products using membranes.     

膜乳化过程研究进展

简要综述了膜乳化过程的机理、操作参数的影响和乳液性质等.操作参数包括膜材料、膜孔径及孔隙率、连续相速率、跨膜压差、乳化剂等.通过控制合适的操作参数,可制备出具有所需尺寸的单分散乳液.     

Analysis of rotating membrane emulsification performance for oil droplet production based on the Taylor vortices approach

Membrane emulsification processes generally employ a cross flow of the continuous phase in order to produce shear stress. Modification of membrane emulsification for the o/w emulsion using the rotating system has been introduced such as the use of stirred cells or a rotating tubular membrane in a stationary liquid. This paper presents an examination of membrane rotation speed on droplet characteristics. The performance of rotating membrane emulsification on o/w droplet size, the coefficient of variation, and size distribution was investigated. In addition, the operated flow regime in the rotating membrane emulsification is addressed. It has been found that overall the droplet size decreased with the increase of membrane rotation speed. The droplet size below the pore size could be produced when operating at a high rotation speed (1500?rpm). The decrease of droplet size was believed due to the action of Taylor vortices on droplet detachment. Analysis of membrane rotation speed proposed that action of Taylor vortices facilitates droplet detachment. Calculation of Taylor numbers (having a value of 0–629) confirmed that the rotating membrane emulsification produced laminar flow with vortices.     

Synthesis of composite emulsions and complex foams with the use of microfluidic flow-focusing devices

A method is described for the formation of stable, composite aqueous emulsions of 1) combinations of distinct families of bubbles of nitrogen, 2) combinations of distinct families of droplets of an organic fluid (either perfluoro(methyl)decalin or hexadecane), and 3) combinations of bubbles and droplets. A system of two or three microfluidic flow-focusing units is coupled to a single outlet channel. The composite emulsions can be precisely tuned, both in their composition and in the number fraction of components--either bubbles or droplets--of different types. The use of microfluidic technology, with closely coupled flow-focusing units, guarantees that the emulsions are mixed locally at a controlled local stoichiometry. The emulsions self-assemble in a nonequilibrium process to form a wide variety of highly organized periodic lattices.     

Influences of combustion improver content and motionless time on the stability of two-phase emulsions

The heavy molecular bonds of liquid fuels can be broken with the assistance of a nitromethane fuel additive by virtue of its explosive and flammable properties to obtain greater heat release. Because of the immiscibility between nitromethane and petro-diesel, two-phase emulsions of nitromethane dispersed in the oil phase of a mixture of diesel and biodiesel were prepared. The experimental results show that microwave irradiation produced an emulsion with a larger number of dispersed nitromethane droplets in the continuous oil mixture, a smaller mean droplet size, and lower turbidity than magnetic stirring, and thus was a better method for preparation of the two-phase emulsion. The increase in the nitromethane weight fraction increased the number of dispersed nitromethane droplets and the emulsion turbidity. In addition, allowing the emulsion preparation to remain motionless for a longer period of time after either method resulted in an obvious reduction in the emulsification stability (ES).     

乳状液制备新工艺——膜乳化过程实验研究

用膜乳化系统制备了Ｏ／Ｗ型乳状液,考察了乳化时间、平均膜孔径、壁面剪应力、膜两侧压差和乳化剂等因素对乳化效果的影响．实验显示,分散相液滴平均直径不随乳化时间而变化;在此条件下,该直径约是膜平均孔径的５～１２倍．随着连续相一侧壁面剪应力的增大液滴平均直径减小,但当壁面剪应力大到一定值后,减小的幅度变得很小．增大膜平均孔径和膜两侧压差都将增加分散相透过膜的通量．此外,乳化剂分子的吸附速度越快,分散相液滴平均直径越小．     

Forming Sticky Droplets from Slippery Polymer Zwitterions

Polymer zwitterions are generally regarded as hydrophilic and repellant or “slippery” materials. Here, a case is described in which the polymer zwitterion structure is tailored to decrease water solubility, stabilize emulsion droplets, and promote interdroplet adhesion. Harnessing the upper critical solution temperature of sulfonium‐ and ammonium‐based polymer zwitterions in water, adhesive droplets are prepared by adding organic solvent to an aqueous polymer solution at elevated temperature, followed by agitation to induce emulsification. Droplet aggregation is observed as the mixture cools. Variation of salt concentration, temperature, polymer concentration, and polymer structure modulates these interdroplet interactions, resulting in distinct changes in emulsion stability and fluidity. Under attractive conditions, emulsions encapsulating 50–75% oil undergo gelation. By contrast, emulsions prepared under conditions where droplets are nonadhesive remain fluid and, for oil fractions exceeding 0.6, coalescence is observed. The uniquely reactive nature of the selected zwitterions allows their in situ modification and affords a route to chemically trigger deaggregation and droplet dispersion. Finally, experiments performed in a microfluidic device, in which droplets are formed under conditions that either promote or suppress adhesion, confirm the salt‐responsive character of these emulsions and the persistence of adhesive interdroplet interactions under flow.     

Preparation of monodisperse W/O emulsions using a stainless-steel microchannel emulsification chip

In this study, hydrophobically treated stainless-steel microchannel (MC) array chips were used for preparing monodisperse W/O emulsions. A water-saturated decane containing 5?wt% tetraglycerin monolaurate condensed ricinoleic acid ester was used as the continuous phase. A Milli-Q water containing 5?wt% polyethylene glycol (molecular weight 20,000) and 5?wt% NaCl was used as the dispersed phase. The stainless-steel MC array chips used for MC emulsification had a sufficiently high contact angle of the dispersed phase to their surface in the continuous phase. The resultant uniform-sized aqueous droplets with a coefficient of variation of <5% had average diameters of 100–300?µm, depending on the MC cross-sectional size. The maximum productivity of uniform-sized aqueous droplets reached higher than 1?mL?h^?1. The difference in the critical capillary number of the dispersed phase that flows in a 100-µm depth MC was 1.5 times greater than that in a 30-µm depth MC.     

氧氟沙星PLGA微囊的制备、表征和影响包封率的因素

氧氟沙星在眼科临床广泛应用,本研究以氧氟沙星作为模型药物,采用水/油/水(w/o/w)的复乳化和溶剂扩散技术制备氧氟沙星聚乳酸-聚乙醇酸(PLGA)微囊,对影响包封率的工艺参数如药物浓度、PLGA使用量、初乳复乳的搅拌速率进行研究,并对微囊的粒径、表面电位和表面形态的理化性能进行了表征。测试结果表明,根据优化工艺制备的氧氟沙星PLGA微囊的平均粒径511.9±14.6nm,zeta电位-17.97±0.80mV,包封率54.2%,载药量1.94%。包封率随PLGA使用量、初乳搅拌速率的增加而上升,随内水相药物体积和浓度的增加而下降。通过优化的水/油/水(w/o/w)复乳化和溶剂扩散技术制备氧氟沙星PLGA载药微囊的粒度分布窄,载药量和包封率适中,具有较好的临床应用前景。     

双乳液体系在微胶囊制备中的应用研究进展

双乳液是一类多重乳状液体系,它具有保护物质并且可以控制这些物质从一个相释放到另一个相的能力。近年来这类乳液体系与传统微胶囊制备方法的结合在药物输送(如抗癌药物、激素等)、食品等领域得到了一定的应用,解决了传统微胶囊制备方法无法有效封装高度水溶性物质等问题。基于此,综述了双乳液体系与微胶囊制备方法结合延伸出的一些新方法,包括双乳液-复凝聚法、复乳溶剂挥发法以及膜乳化复乳法等;同时,评述了影响双乳液体系制备微胶囊的各种因素,展望了双乳液体系在农药微胶囊制备中的应用前景。     

Fabrication of advanced particles and particle-based materials assisted by droplet-based microfluidics

Recent advances in the fabrication of complex particles and particle-based materials assisted by droplet-based microfluidics are reviewed. Monodisperse particles with expected internal structures, morphologies, and sizes in the range of nanometers to hundreds of micrometers have received a good deal of attention in recent years. Due to the capability of generating monodisperse emulsions and of executing precise control and operations on the suspended droplets inside the microchannels, droplet-based microfluidic devices have become powerful tools for fabricating complex particles with desired properties. Emulsions and multiple-emulsions generated in the microfluidic devices can be composed of a variety of materials including aqueous solutions, gels, polymers and solutions containing functional nanoparticles. They are ideal microreactors or fine templates for synthesizing advanced particles, such as polymer particles, microcapsules, nanocrystals, and photonic crystal clusters or beads by further chemical or physical operations. These particles are promising materials that may be applicable for many fields, such as photonic materials, drug delivery systems, and bio-analysis. From simple to complex, from spherical to nonspherical, from polymerization and reaction crystallization to self-assembly, this review aims to help readers be aware of the many aspects of this field.     

Monodisperse, Submicrometer Droplets via Condensation of Microfluidic-Generated Gas Bubbles

Microfluidics (MFs) can produce monodisperse droplets with precise size control. However, the synthesis of monodisperse droplets much smaller than the minimum feature size of the microfluidic device (MFD) remains challenging, thus limiting the production of submicrometer droplets. To overcome the minimum micrometer-scale droplet sizes that can be generated using typical MFDs, the droplet material is heated above its boiling point (bp), and then MFs is used to produce monodisperse micrometer-scale bubbles (MBs) that are easily formed in the size regime where standard MFDs have excellent size control. After MBs are formed, they are cooled, condensing into dramatically smaller droplets that are beyond the size limit achievable using the original MFD, with a size decrease corresponding to the density difference between the gas and liquid phases of the droplet material. Herein, it is shown experimentally that monodisperse, submicrometer droplets of predictable sizes can be condensed from a monodisperse population of MBs as generated by MFs. Using perfluoropentane (PFP) as a representative solvent due to its low bp (29.2 °C), it is demonstrated that monodisperse PFP MBs can be produced at MFD temperatures >3.6 °C above the bp of PFP over a wide range of sizes (i.e., diameters from 2 to 200 μm). Independent of initial size, the generated MBs shrink rapidly in size from about 3 to 0 °C above the bp of PFP, corresponding to a phase change from gas to liquid, after which they shrink more slowly to form fully condensed droplets with diameters 5.0 ± 0.1 times smaller than the initial size of the MBs, even in the submicrometer size regime. This new method is versatile and flexible, and may be applied to any type of low-bp solvent for the manufacture of different submicrometer droplets for which precisely controlled dimensions are required.     

Development of flurbiprofen-loaded nanoparticles with a narrow size distribution using sucrose

Objective: A novel flurbiprofen-loaded nanoemulsion which gave uniform emulsion droplets with a narrow size distribution was previously reported to be prepared using membrane emulsification method. The purpose of this study is to develop a novel flurbiprofen-loaded nanoparticle with a narrow size distribution and improved bioavailability.

Method: The nanoparticle was prepared by solidifying nanoemulsion using sucrose as a carrier via spray drying method. Its physicochemical properties were investigated using SEM, DSC and PXRD. Furthermore, dissolution and bioavailability in rats were evaluated compared to a flurbiprofen-loaded commercial product.

Results: The flurbiprofen-loaded nanoparticles with flurbiprofen/sucrose/surfactant mixture (1/20/2, weight ratio) gave good solidification and no stickiness. They associated with about 70?000-fold improved drug solubility and had a mean size of about 300 nm with a narrow size distribution. Flurbiprofen was present in a changed amorphous state in these nanoparticles. Moreover, the nanoparticles gave significantly shorter T_max, and higher AUC and C_max of the drug compared to the commercial product (p?0.05). In particular, they showed about nine-fold higher AUC of the drug than did the commercial product

Conclusion: These flurbiprofen-loaded nanoparticles prepared with sucrose by the membrane emulsification and spray drying method would be a potential candidate for orally delivering poorly water-soluble flurbiprofen with enhanced bioavailability.     

Gravity-driven microfluidic particle sorting device with hydrodynamic separation amplification

This paper describes a simple microfluidic sorting system that can perform size profiling and continuous mass-dependent separation of particles through combined use of gravity (1 g) and hydrodynamic flows capable of rapidly amplifying sedimentation-based separation between particles. Operation of the device relies on two microfluidic transport processes: (i) initial hydrodynamic focusing of particles in a microchannel oriented parallel to gravity and (ii) subsequent sample separation where positional difference between particles with different mass generated by sedimentation is further amplified by hydrodynamic flows whose streamlines gradually widen out due to the geometry of a widening microchannel oriented perpendicular to gravity. The microfluidic sorting device was fabricated in poly(dimethylsiloxane), and hydrodynamic flows in microchannels were driven by gravity without using external pumps. We conducted theoretical and experimental studies on fluid dynamic characteristics of laminar flows in widening microchannels and hydrodynamic amplification of particle separation. Direct trajectory monitoring, collection, and post-analysis of separated particles were performed using polystyrene microbeads with different sizes to demonstrate rapid (<1 min) and high-purity (>99.9%) separation. Finally, we demonstrated biomedical applications of our system by isolating small-sized (diameter <6 microm) perfluorocarbon liquid droplets from polydisperse droplet emulsions, which is crucial in preparing contrast agents for safe, reliable ultrasound medical imaging, tracers for magnetic resonance imaging, or transpulmonary droplets used in ultrasound-based occlusion therapy for cancer treatment. Our method enables straightforward, rapid, real-time size monitoring and continuous separation of particles in simple stand-alone microfabricated devices without the need for bulky and complex external power sources. We believe that this system will provide a useful tool to separate colloids and particles for various analytical and preparative applications and may hold potential for separation of cells or development of diagnostic tools requiring point-of-care sample preparation or testing.     

Sequential Coalescence Enabled Two‐Step Microreactions in Triple‐Core Double‐Emulsion Droplets Triggered by an Electric Field

Advances in microfluidic emulsification have enabled the generation of exquisite multiple‐core droplets, which are promising structures to accommodate microreactions. An essential requirement for conducting reactions is the sequential coalescence of the multiple cores encapsulated within these droplets, therefore, mixing the reagents together in a controlled sequence. Here, a microfluidic approach is reported for the conduction of two‐step microreactions by electrically fusing three cores inside double‐emulsion droplets. Using a microcapillary glass device, monodisperse water‐in‐oil‐in‐water droplets are fabricated with three compartmented reagents encapsulated inside. An AC electric field is then applied through a polydimethylsiloxane chip to trigger the sequential mixing of the reagents, where the precise sequence is guaranteed by the discrepancy of the volume or conductivity of the inner cores. A two‐step reaction in each droplet is ensured by two times of core coalescence, which totally takes 20–40 s depending on varying conditions. The optimal parameters of the AC signal for the sequential fusion of the inner droplets are identified. Moreover, the capability of this technique is demonstrated by conducting an enzyme‐catalyzed reaction used for glucose detection with the double‐emulsion droplets. This technique should benefit a wide range of applications that require multistep reactions in micrometer scale.     

Synthesis of monodisperse, covalently cross-linked, degradable "smart" microgels using microfluidics

The development of a robust method for the synthesis of highly monodisperse microgels cross-linked with degradable covalent bonds offers the potential for fabricating microgels with the highly controllable porosities, cell interactions, and degradation half-lives required for biomedical applications. A microfluidic chip is designed that enables the on-chip mixing and emulsification of two reactive polymer solutions (hydrazide and aldehyde-functionalized carbohydrates) to form monodisperse, hydrazone cross-linked microgels in the size range of ≈40-100 μm. The device can be run continuously for at least 30 h without a significant drift in particle size. The resulting microgels have a homogeneous bulk composition and can swell and deswell as the solvent conditions change in predictable ways based on the chemistry of the reactive polymers used, thereby enabling improved control over both the chemistry and morphology of the resulting microgels relative to other reported approaches. The in situ gelation chemistry used facilitates rapid microgel formation within the droplets without requiring the use of UV light or heating to initiate polymerization, thus making this approach of particular potential utility in cell encapsulation or drug delivery (as demonstrated).     

Microfluidic fabrication of asymmetric giant lipid vesicles

We have developed a microfluidic technology for the fabrication of compositionally asymmetric giant unilamellar vesicles (GUVs). The vesicles are assembled in two independent steps. In each step, a lipid monolayer is formed at a water-oil interface. The first monolayer is formed inside of a microfluidic device with a multiphase droplet flow configuration consisting of a continuous oil stream in which water droplets are formed. These droplets are dispensed into a vessel containing a layer of oil over a layer of water. The second lipid monolayer is formed by transferring the droplets through this second oil-water interface by centrifugation. By dissolving different lipid compositions in the different oil phases, the composition of each leaflet of the resulting lipid bilayer can be controlled. We have demonstrated membrane asymmetry by showing differential fluorescence quenching of labeled lipids in each leaflet and by demonstrating that asymmetric GUVs will bind an avidin-coated surface only when biotinylated lipids are targeted to the outer leaflet. In addition, we have demonstrated the successful asymmetric targeting of phosphatidylserine lipids to each leaflet, producing membranes with a biomimetic and physiologically relevant compositional asymmetry.     

Osmocapsules for Direct Measurement of Osmotic Strength

Monodisperse microcapsules with ultra‐thin membranes are microfluidically designed to be highly sensitive to osmotic pressure, thereby providing a tool for the direct measurement of the osmotic strength. To make such osmocapsules, water‐in‐oil‐in‐water double‐emulsion drops with ultra‐thin shells are prepared as templates through emulsification of core–sheath biphasic flow in a capillary microfluidic device. When photocurable monomers are used as the oil phase, the osmocapsules are prepared by in‐situ photopolymerization of the monomers, resulting in semipermeable membranes with a relatively large ratio of membrane thickness to capsule radius, approximately 0.02. These osmocapsules are buckled by the outward flux of water when they are subjected to a positive osmotic pressure difference above 125 kPa. By contrast, evaporation‐induced consolidation of middle‐phase containing polymers enables the production of osmocapsules with a small ratio of membrane thickness to capsule radius of approximately 0.002. Such an ultra‐thin membrane with semi‐permeability makes the osmocapsules highly sensitive to osmotic pressure; a positive pressure as small as 12.5 kPa induces buckling of the capsules. By employing a set of distinct osmocapsules confining aqueous solutions with different osmotic strengths, the osmotic strength of unknown solutions can be estimated through observation of the capsules that are selectively buckled. This approach provides the efficient measurement of the osmotic strength using only a very small volume of liquid, thereby providing a useful alternative to other measurement methods which use complex setups. In addition, in‐vivo measurement of the osmotic strength can be potentially accomplished by implanting these biocompatible osmocapsules into tissue, which is difficult to achieve using conventional methods.     

微流控技术构建单分散微囊膜的研究新进展

单分散微囊膜在控制释放领域显示出许多优势而备受重视,而近期出现的微流控技术为制备单分散微囊膜提供了可靠的新方法.综述了微流控技术构建单分散微囊膜的研究新进展,着重介绍了微流控技术制备基于乙基纤维素、壳聚糖和海藻酸钙等生物相容性材料的单分散微囊膜以及基于聚N-异丙基丙烯酰胺的温敏型和离子识别型的单分散微囊膜的研究现状.