[show abstract][hide abstract] ABSTRACT: Incorporation of locally produced signaling molecules into cell-derived vesicles may serve as an endogenous mediator delivery system. We recently reported that levels alpha-2-macroglobulin (A2MG)-containing microparticles are elevated in plasma from patients with sepsis. Herein, we investigated the immunomodulatory actions of A2MG containing microparticles during sepsis. Administration of A2MG-enriched (A2MG-E)-microparticles to mice with microbial sepsis protected against hypothermia, reduced bacterial titers, elevated immunoresolvent lipid mediator levels in inflammatory exudates and reduced systemic inflammation. A2MG-E microparticles also enhanced survival in murine sepsis, an action lost in mice transfected with siRNA for LRP1, a putative A2MG receptor. In vitro, A2MG was functionally transferred onto endothelial cell plasma membranes from microparticles, augmenting neutrophil-endothelial adhesion. A2MG also modulated human leukocyte responses: enhanced bacterial phagocytosis, reactive oxygen species production, cathelicidin release, prevented endotoxin induced CXCR2 downregulation and preserved neutrophil chemotaxis in the presence of LPS. A significant association was also found between elevated plasma levels of A2MG-containing microparticles and survival in human sepsis patients. Taken together, these results identify A2MG enrichment in microparticles as an important host protective mechanism in sepsis.
EMBO Molecular Medicine 12/2013; · 7.80 Impact Factor
[show abstract][hide abstract] ABSTRACT: Abstract Biodegradable hollow capsules filled with fluorescently labelled bovine serum albumin (BSA) as a model drug were prepared via layer-by-layer (LbL) self-assembly of type-I collagen (COL) and hyaluronic acid (HA) using calcium carbonate micro-particles and co-precipitation method. Capsules loaded with fluorescein isothiocyanate (FITC)-BSA, tetramethylrhodamin isothiocyanate (TRITC)-BSA or Alex-Fluor-488-BSA, respectively, were characterised before and after core removal using Confocal Laser Scanning Microscopy (CLSM), whilst the morphologies of individual hollow capsules were assessed using Atomic Force Microscopy (AFM). The sustained release of the encapsulated FITC-BSA protein was attained using enzymatic degradation of the capsule shells by collagenase. The released profile of the fluorescently-labelled BSA indicated that it could be successfully controlled by modulating the number of layers and/or by collagen crosslinking either before or after the capsule's assembly.
Journal of Microencapsulation 10/2013; · 1.57 Impact Factor
[show abstract][hide abstract] ABSTRACT: By introducing UV-sensitive chemical groups causing different potential response as building blocks, fabricated LbL capsules can be endowed with dual UV-responsive properties in specific layers. One block is responsible for fast capsule sealing and the other for longer term capsule swelling and rupture. Therefore, the multifunction of these capsules could be activated selectively when exposed to external UV light with suitable wavelengths. In this work, dual-functional complex microcapsules (PDADMAC/PAZO)4-(DAR/Nafion)2 containing both diazonium and aozbenzene groups were proposed as clear examples to realize a time-dependent UV response for successive encapsulation and release. Upon exposure to UV light, the DAR/Nafion layers underwent a rapid in situ cross-linking and hence to seal the capsule shells through diazonium-related photolysis. Then further gradual shell swelling was followed by realignment of azobenzene molecules in PDADMAC/PAZO layers. Fluorescent polymers were consequently studied as cargo substances. Results indicated that continuous UV light triggered rapid cargo encapsulation over minutes time scale and gradual release with continuous irradiation over hours.
[show abstract][hide abstract] ABSTRACT: Layer-by-layer assembled shells are prospective candidates for encapsulation, stabilization, storage, and release of fragrances. A shell comprising four alternative layers of a protein and a polyphenol is employed to encapsulate the dispersed phase of a fragrance-containing oil-in-water emulsion. The model fragrance used in this work consists of 10 ingredients, covering a range of typically employed aroma molecules, all premixed in equal mass and with sunflower oil acting as the base. The encapsulated emulsion is stable after 2 months of storage at 4 °C as revealed by static light scattering and confocal laser scanning microscopy. Gas chromatography/mass spectrometry data show that the encapsulation efficiency of 8 out of 10 fragrance ingredients depends on the water solubility: the less water-soluble an ingredient, the more of it is encapsulated. The amount of these fragrance ingredients remaining encapsulated decreases linearly upon emulsion incubation at 40 °C and the multilayer shell does not hinder their release. The other two fragrance ingredients having the lowest saturation vapor pressure demonstrate sustained release over 5 days of incubation at 40 °C. The composition of released fragrance remains almost constant over 3 days of incubation, upon further incubation it becomes enriched with these two ingredients when others start to be depleted.
[show abstract][hide abstract] ABSTRACT: Layer-by-layer assembled microcapsules have the potential to be versatile cell delivery systems incorporating multiple activities and functions. However, it is necessary to determine the influence that different capsule locations have on activity of bioactive molecules in order to optimise delivery and for generation of multifunctional capsules. In this study we examine the influence that locating the bioluminescent enzyme luciferase in different microcapsule locations has on activity in intact synthetic and biodegradable microcapsules before and after cell delivery as well as its susceptibility to protease degradation. We also examine the effect of microcapsule position on cell transfection with plasmid DNA. Based on the findings of experiments in this study we also demonstrate co-delivery of luciferase protein and plasmid DNA encoding a fluorescent protein from two different locations within the same microcapsule. Our studies confirm that, the core, subouter layer, and outer layer are optimal for cell delivery but these positions offer least protection from protease activity. By contrast middle layer molecules remain entangled with capsule layers preventing their release which is inefficient for cell delivery but this provides better protection from protease degradation. The findings of this study will enable more rationale layer-by-layer assembly of microcapsules containing biologically active molecules for cell delivery and aid in the generation of multifunctional microcapsules.
Journal of Controlled Release 08/2013; · 7.63 Impact Factor
[show abstract][hide abstract] ABSTRACT: Novel microcapsule systems containing UV-responsive diazonium groups were fabricated as micro-containers for cargo substance encapsulation by using LbL assembly technique. Upon direct exposure to UV light with a wavelength of approximately 380 nm, the diazonium groups of DAR rapidly reacted with sulfonate or diazo-sulfonate groups of counterpart polyelectrolytes, which converted electrostatic interactions to covalent bonds, demonstrating as effective in-situ crosslinking within multilayers via photolysis. Such chemical transition eliminated the paired ionic groups, therefore generating more hydrophobic multilayer shells, offering a unique approach to seal the porous polyelectrolyte capsule shells. Fluorescent molecule RhB was consequently studied as a typical example for small molecule encapsulation. Results indicated that the dye was remarkably retained within the microcapsules after UV-triggered capsule shell sealing.
[show abstract][hide abstract] ABSTRACT: Calcium, cadmium and manganese carbonate crystals were used as core material for fabrication of hollow polyelectrolyte capsules by means of the Layer-by-Layer assembly. The use of inorganic templates is a way of fabrication of clean capsules, which is essential for basic research and is a significant step towards their biocompatibility. The ways of particle and capsule fabrication and characterization are described. Scanning electron microscopy (SEM)-Energy dispersive X-ray spectroscopy (EDX) measurements proved the purity of the hollow capsules from the core material. The capsules obtained were characterized by scanning force microscopy, and confocal fluorescence microscopy.
[show abstract][hide abstract] ABSTRACT: Layer-by-layer assembled microcapsules have potential applications as delivery and biosensing systems which make them attractive tools for use in various aspects of nanomedicine. We examined the effect of microcapsule location on activity of the bioluminescent enzyme luciferase in both intact capsules and following cell uptake. In intact capsules the rate of reaction of luciferase was greatest for luciferase in the outer layer and least in the core. Following cell uptake, luciferase in the outer layer was rapidly reactive, a similar rate of reaction and activity was observed for luciferase placed in capsule interior (core). By contrast there was minimal activity detected when microcapsules with luciferase sandwiched between polyelectrolytes in a middle layer were delivered to cells. This study informs us of the availability of bioactive molecules located in different positions within microcapsules and will enable better microcapsule construction in line with the intended application, particularly delivery of functional proteins to cells.
[show abstract][hide abstract] ABSTRACT: Patterned arrays of light-responsive microchambers are suggested as candidates for site-specific release of chemicals in small and precisely defined quantities on demand. A composite film is made of poly(allylammonium)-poly(styrene sulfonate) multilayers and gold nanoparticles incorporated between subsequent stacks of polyelectrolytes. The film shaped as microchambers is loaded with colloid particles or oil-soluble molecules. The microchambers are sealed onto a glass slide precoated with an adhesive poly(diallyldimethylammonium)-poly(styrene sulfonate) multilayer film. A focused laser beam is used for remote addressing the individual microchambers and site-specific release of the loaded cargo.
[show abstract][hide abstract] ABSTRACT: The concept of enzyme-assisted substrate sensing based on use of fluorescent markers to detect the products of enzymatic reaction has been investigated by fabrication of micron-scale polyelectrolyte capsules containing enzymes and dyes in one entity. Microcapsules approximately 5 μm in size entrap glucose oxidase or lactate oxidase, with peroxidase, together with the corresponding markers Tris(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dichloride (Ru(dpp)) complex and dihydrorhodamine 123 (DHR123), which are sensitive to oxygen and hydrogen peroxide, respectively. These capsules are produced by co-precipitation of calcium carbonate particles with the enzyme followed by layer-by-layer assembly of polyelectrolytes over the surface of the particles and incorporation of the dye in the capsule interior or in the multilayer shell. After dissolution of the calcium carbonate the enzymes and dyes remain in the multilayer capsules. In this study we produced enzyme-containing microcapsules sensitive to glucose and lactate. Calibration curves based on fluorescence intensity of Ru(dpp) and DHR123 were linearly dependent on substrate concentration, enabling reliable sensing in the millimolar range. The main advantages of using these capsules with optical recording is the possibility of building single capsule-based sensors. The response from individual capsules was observed by confocal microscopy as increasing fluorescence intensity of the capsule on addition of lactate at millimolar concentrations. Because internalization of the micron-sized multi-component capsules was feasible, they could be further optimized for in-situ intracellular sensing and metabolite monitoring on the basis of fluorescence reporting.
Analytical and Bioanalytical Chemistry 09/2012; · 3.66 Impact Factor
[show abstract][hide abstract] ABSTRACT: Microcapsules composed of weak polyelectrolytes modified with UV-responsive benzophenone (BP) groups were fabricated by the layer-by-layer (LbL) technique. Being exposed to UV lights, capsules shrunk in the time course of minutes at irradiation intensity of 5 mW/cm(2). The shrinkage adjusted the capsule permeability, providing a novel way to encapsulate fluorescence-labeled dextran molecules without heating. Cross-linking within the capsule shells based on hydrogen abstraction via excited benzophenone units by UV showed a reliable and swift approach to tighten and stabilize the capsule shell without losing the pH-responsive properties of the weak polyelectrolyte multilayers.
[show abstract][hide abstract] ABSTRACT: Basic fibroblast growth factor (FGF2) is an important protein for cellular activity and highly vulnerable to environmental conditions. FGF2 protected by heparin and bovine serum albumin was loaded into the microcapsules by a coprecipitation-based layer-by-layer encapsulation method. Low cytotoxic and biodegradable polyelectrolytes dextran sulfate and poly-L-arginine were used for capsule shell assembly. The shell thickness-dependent encapsulation efficiency was measured by enzyme-linked immunosorbent assay. A maximum encapsulation efficiency of 42% could be achieved by microcapsules with a shell thickness of 14 layers. The effects of microcapsule concentration and shell thickness on cytotoxicity, FGF2 release kinetics, and L929 cell proliferation were evaluated in vitro. The advantage of using microcapsules as the carrier for FGF2 controlled release for enhancing L929 cell proliferation was analyzed.
[show abstract][hide abstract] ABSTRACT: Polyelectrolyte multilayer (PEM) films with array of responsive microchambers are promising candidates for site-specific release of chemicals in small and precisely defined quantities on demand. It requires effective sealing of the microchambers toward a support to prevent leakage of a cargo. In this paper, we study the pressure-induced adhesion of poly(allylammonium)-poly(4-styrenesulfonate) (PAH-PSS) multilayers assembled on different templates toward the poly(4-styrenesulfonate)-poly(diallyldimethylammonium) multilayer. The tensile bond strength increases from 0.4 to 3.5 MPa upon the increase of PAH-PSS bilayers from 10 to 40, if assembled on a silicon template. Weaker tensile bond strength of 0.35 MPa between the PAH-PSS multilayer and a poly(methylmethacrylate) (PMMA) template results in adhesive break at this interface and allows mechanical removal of the template. The successful PEM transfer is demonstrated for templates of various geometrical patterns, while the tensile break of a multilayer film happens for the others.
[show abstract][hide abstract] ABSTRACT: Optical coherence tomography (OCT) is a new, non-invasive, noncontact in vivo imaging technology. We demon-strated that the OCT can be used as a sufficient technique for nanocomposite microcapsule visualization in a liquid medium. As a model system we choose a water/glycerol mixture with viscosity in a variable range from 1.01 to 1.41×10 3 mPa s, in-cluding viscosity of a blood plasma. We have found that to-mography spatial resolution is enough to visualize capsules and their aggregates in liquids and to estimate their concen-tration via two-dimensional (2D) tomography scan analysis. In our experimental conditions microcapsule concentration mea-sured by OCT was 3.9×10 7 cm −3 and this value correlated well with the concentration measured in a counting chamber (1.9×10 7 cm −3). We also demonstrated the possibility to con-trol capsule spatial distribution in glycerol solutions by external magnetic field and determined the dependence of capsule sedi-mentation time on the liquid medium viscosity. 50 dB 40 dB 30 dB 20 dB 10 dB (a) (b) (c) (d) N S O C T Two-dimensional tomographic scans of a cuvette filled with mi-crocapsules suspension in 50% glycerol in water solution c ⃝ 2012 by Astro, Ltd.
[show abstract][hide abstract] ABSTRACT: Light-responsive microcapsules constructed by layer-by-layer self-assembly are used as microcarriers to deliver different macromolecules inside cells. The microcapsules carry the macromolecules as cargo in their cavity, while their walls are modified with agglomerated gold nanoparticles. Microcapsules are incorporated by living cells and are then located in lysosomal compartments. Controlled release of the encapsulated material from the interior of the capsule to the cytosol is possible upon NIR-light irradiation. This is based on local heating of the gold nanoparticles upon NIR light and disruption of the capsule wall, what results on release of encapsulated materials. We illustrate several key advances in controlled release induced by light. First, we demonstrate that capsules can be opened individually, which allows for sequentially releasing cargo from different capsules within one single cell. Second, by using a pH-indicator as cargo the claim of release from the acidic lysosomal compartments to the neutral cytosol is experimentally evident which until now has been only speculated. Third, green fluorescent protein (GFP) is released to the cytosol while retaining its functionality. This demonstrates that proteins can be released without destruction by the local heating. Fourth, GFP is also administered in biodegradable capsules, which leads to a different release mechanism compared to externally triggering for light-responsive microcapsules.
Journal of Controlled Release 12/2011; 159(1):120-7. · 7.63 Impact Factor