Mucin coated protein-polyphenol microcarriers for daidzein delivery.

Daidzein, an isoflavone found abundantly in legumes, may benefit from bypassing upper gut absorption to reach the colon where it can be metabolized into the potent estrogen equol by the gut microbiome. To achieve this, we developed mucin coated protein-tannin multilayer microcarriers. Highly porous functionalized calcium carbonate (FCC) microparticles efficiently absorbed daidzein from a dimethyl sulfoxide solution, with a loading capacity of 21.6 ± 1.8 wt% as measured by ultra-high pressure liquid chromatography - mass spectrometry (UPLC-MS). Daidzein-containing FCC microparticles were then coated with a bovine serum albumin (BSA)-tannin n-layer film terminated with mucin ((BSA-TA)n-mucin) by layer-by-layer deposition from corresponding aqueous solutions followed by FCC decomposition with HCl. Raman spectroscopy confirmed mucin-tannin complexation involving both hydrophobic interactions and hydrogen bonding. The resulting multilayer microcarriers contained 54 wt% of nanocrystalline daidzein as confirmed by X-ray diffraction and UPLC-MS. Preliminary screening of several types of mucin coatings using an in vitro INFOGEST digestion model demonstrated that mucin type III from porcine stomach provided the highest protection against upper intestinal digestion. (BSA-TA)8-mucin and (BSA-TA)4-mucin microcarriers retained 71 ± 16.4% and 68 ± 4.6% of daidzein, respectively, at the end of the small intestinal phase. Mucin-free (BSA-TA)8 retained a lower daidzein amount of 46%. Daidzein release and further conversion into equol were observed during in vitro colonic studies with fecal microbiota from a healthy non-equol-producing donor and Slackia equolifaciens. The developed approach has potential for encapsulating other hydrophobic nutraceuticals or therapeutics, enhancing their bioaccessibility in the colon.

Surface-reacted calcium carbonate microparticles as templates for lactoferrin encapsulation.

Microencapsulation helps to improve bioavailability of a functional whey protein, lactoferrin (Lf), in adults. Herein, we report the Lf loading capacity (LC) and retention efficiency (RE) in the microparticles of surface-reacted calcium carbonate (SRCC) of different types and compare them to those of widely used vaterite microparticles. The LCs and REs are analyzed in connection to the total surface area and the volume of intraparticle pores. The best performing SRCC3 demonstrates Lf LC of 11.00 wt% achieved in a single absorption step and 74% RE after two cycles of washing with deionized water. A much larger surface area of SRCC templates and a lower pH required to release Lf do not affect its antitumor activity in MCF-7 assay. Layer-by-Layer assembly of pepsin-tannic acid multilayer shell around Lf-loaded microparticles followed by acidic decomposition of the inorganic core produces microencapsulated Lf with a yield ~36 times higher than from vaterite templates reported earlier, while the scale of encapsulated Lf production is ~12,000 times larger. In vitro digestion tests demonstrate the protection of ~65% of encapsulated Lf from gastric digestion. The developed capsules are prospective candidates for functional foods fortified with Lf.

Arrays of Biocompatible and Mechanically Robust Microchambers Made of Protein-Polyphenol-Clay Multilayer Films.

There is a growing demand for biocompatible and mechanically robust arrays of microcompartments loaded with minute amounts of active substances for sensing or controlled release applications. Here we report on a novel biocompatible composite material, protein-polyphenol-clay (PPC) multilayer film. The material is shown to be strong enough to make robust microchambers retaining the shape and dimensions of truncated square pyramids. We study the mechanical properties and biocompatibility of the PPC microchambers and compare them to those made of synthetic polyelectrolyte multilayer film, poly(styrenesulfonate)-poly(allylammonium) (PSS-PAH). The mechanical properties of the microchambers were characterized under uniaxial compression using nanoindentation with a flat-punch tip. The effective Young's modulus of PPC microchambers, 166 ± 53 MPa, is found to be lower than that of PSS-PAH microchambers, 245 ± 52 MPa. However, the capacity to elastically absorb the energy of the former, 2.4 ± 1.0 MPa, is marginally higher than of the latter, 2.0 ± 1.3 MPa. Arrays of microchambers were sealed onto a polyethylene film, loaded with a model oil-soluble drug, and their biocompatibility was tested using an ex vivo 3D human skin reconstruct model. We found no evidence for toxicity with the PPC microchambers; however, PSS-PAH microchambers stimulated reduced cell density in the epidermis and significantly affected epidermal-dermal attachment. Both materials do not alter skin cell proliferation but affect skin cell differentiation. We interpret that rather than affecting epidermal barrier function, these data suggest the applied plastic films with microchamber arrays affect transpiration, normoxia, and moisture exchange.

Origin of High Thermoelectric Performance in Earth-Abundant Phosphide-Tetrahedrite.

Phosphide-based thermoelectrics are a relatively less studied class of compounds, primarily due to the presence of light elements, which result in high thermal conductivity and inherent stability problems. In this work, we present a stable phosphide-tetrahedrite, Ag6Ge10P12, which possesses the highest zT (∼0.7) among all known phosphides at intermediate temperatures (750 K). We examine the intrinsic electronic and thermal transport properties of this compound by expressing the transport properties in terms of weighted mobility (µW), transport coefficient (σE0), and material quality factor (B), from which we are able to elucidate that the origin of its high zT can be attributed to the platelike Fermi surface and high level of band multiplicity related to its complex band structure. Finally, we discuss the origin of the low lattice thermal conductivity observed in this compound using experimental sound velocity, elastic properties, and Debye-Callaway model, thus laying the foundation for similar stable phosphides as potentially earth-abundant and nontoxic intermediate-temperature thermoelectric materials.

Polyelectrolyte-Graphene Oxide Multilayer Composites for Array of Microchambers which are Mechanically Robust and Responsive to NIR Light.

Development of composite polymer/graphene oxide (GO) materials attracts significant attention due to their unique properties. In this work, highly ordered arrays of hollow microchambers made of composite polyelectrolyte/GO multilayers (PEGOMs) are successfully fabricated via layer-by-layer assembly on sacrificial or sustainable templates having imprinted patterns of microwells on their surface. Mechanical and optical properties of PEGOMs are studied by nanoindentation and near-infrared (NIR) absorption spectroscopy. Incorporation of three GO layers in between the polyelectrolyte multilayer stacks increases Young's modulus and critical stress of the microchambers by a factor of 5.6 and 2.6, respectively. Optical density of this PEGOM film is found to decrease gradually from 0.14 at λ = 800 nm to 0.06 at λ = 1500 nm. Remote opening of PEGOM microchambers with NIR laser beam is also demonstrated. One of the possible applications of the developed structures includes micropackaging and delivery systems in biological tissues with remote triggering.

Protein-tannic acid multilayer films: A multifunctional material for microencapsulation of food-derived bioactives.

The benefits of various functional foods are often negated by stomach digestion and poor targeting to the lower gastrointestinal tract. Layer-by-Layer assembled protein-tannic acid (TA) films are suggested as a prospective material for microencapsulation of food-derived bioactive compounds. Bovine serum albumin (BSA)-TA and pepsin-TA films demonstrate linear growth of 2.8±0.1 and 4.2±0.1nm per bi-layer, correspondingly, as shown by ellipsometry. Both multilayer films are stable in simulated gastric fluid but degrade in simulated intestinal fluid. Their corresponding degradation constants are 0.026±0.006 and 0.347±0.005nm-1min-1. Milk proteins possessing enhanced adhesion to human intestinal surface, Immunoglobulin G (IgG) and ß-Lactoglobulin (BLG), are explored to tailor targeting function to BSA-TA multilayer film. BLG does not adsorb onto the multilayer while IgG is successfully incorporated. Microcapsules prepared from the multilayer demonstrate 2.7 and 6.3 times higher adhesion to Caco-2 cells when IgG is introduced as an intermediate and the terminal layer, correspondingly. This developed material has a great potential for oral delivery of numerous active food-derived ingredients.

Formulation for Oral Delivery of Lactoferrin Based on Bovine Serum Albumin and Tannic Acid Multilayer Microcapsules.

Lactoferrin (Lf) has considerable potential as a functional ingredient in food, cosmetic and pharmaceutical applications. However, the bioavailability of Lf is limited as it is susceptible to digestive enzymes in gastrointestinal tract. The shells comprising alternate layers of bovine serum albumin (BSA) and tannic acid (TA) were tested as Lf encapsulation system for oral administration. Lf absorption by freshly prepared porous 3 µm CaCO3 particles followed by Layer-by-Layer assembly of the BSA-TA shells and dissolution of the CaCO3 cores was suggested as the most efficient and harmless Lf loading method. The microcapsules showed high stability in gastric conditions and effectively protected encapsulated proteins from digestion. Protective efficiency was found to be 76 ± 6% and 85 ± 2%, for (BSA-TA)4 and (BSA-TA)8 shells, respectively. The transit of Lf along the gastrointestinal tract (GIT) of mice was followed in vivo and ex vivo using NIR luminescence. We have demonstrated that microcapsules released Lf in small intestine allowing 6.5 times higher concentration than in control group dosed with the same amount of free Lf. Significant amounts of Lf released from microcapsules were then absorbed into bloodstream and accumulated in liver. Suggested encapsulation system has a great potential for functional foods providing lactoferrin.

Flexible palladium-based H2 sensor with fast response and low leakage detection by nanoimprint lithography.

Flexible palladium-based H2 sensors have a great potential in advanced sensing applications, as they offer advantages such as light weight, space conservation, and mechanical durability. Despite these advantages, the paucity of such sensors is due to the fact that they are difficult to fabricate while maintaining excellent sensing performance. Here, we demonstrate, using direct nanoimprint lithography of palladium, the fabrication of a flexible, durable, and fast responsive H2 sensor that is capable of detecting H2 gas concentration as low as 50 ppm. High resolution and high throughput patterning of palladium gratings over a 2 cm × 1 cm area on a rigid substrate was achieved by heat-treating nanoimprinted palladium benzyl mercaptide at 250 °C for 1 h. The flexible and robust H2 sensing device was fabricated by subsequent transfer nanoimprinting of these gratings into a polycarbonate film at its glass transition temperature. This technique produces flexible H2 sensors with improved durability, sensitivity, and response time in comparison to palladium thin films. At ambient pressure and temperature, the device showed a fast response time of 18 s at a H2 concentration of 3500 ppm. At 50 ppm concentration, the response time was found to be 57 s. The flexibility of the sensor does not appear to compromise its performance.

Metal hierarchical patterning by direct nanoimprint lithography.

Three-dimensional hierarchical patterning of metals is of paramount importance in diverse fields involving photonics, controlling surface wettability and wearable electronics. Conventionally, this type of structuring is tedious and usually involves layer-by-layer lithographic patterning. Here, we describe a simple process of direct nanoimprint lithography using palladium benzylthiolate, a versatile metal-organic ink, which not only leads to the formation of hierarchical patterns but also is amenable to layer-by-layer stacking of the metal over large areas. The key to achieving such multi-faceted patterning is hysteretic melting of ink, enabling its shaping. It undergoes transformation to metallic palladium under gentle thermal conditions without affecting the integrity of the hierarchical patterns on micro- as well as nanoscale. A metallic rice leaf structure showing anisotropic wetting behavior and woodpile-like structures were thus fabricated. Furthermore, this method is extendable for transferring imprinted structures to a flexible substrate to make them robust enough to sustain numerous bending cycles.

Direct nanoimprint lithography of Al2O3 using a chelated monomer-based precursor.

Nanostructuring of Al2O3 is predominantly achieved by the anodization of aluminum film and is limited to obtaining porous anodized aluminum oxide (AAO). One of the main restrictions in developing approaches for direct fabrication of various types of Al2O3 patterns, such as lines, pillars, holes, etc, is the lack of a processable aluminum-containing resist. In this paper, we demonstrate a stable precursor prepared by reacting aluminum tri-sec-butoxide with 2-(methacryloyloxy)ethyl acetoacetate, a chelating monomer, which can be used for large area direct nanoimprint lithography of Al2O3. Chelation in the precursor makes it stable against hydrolysis whilst the presence of a reactive methacrylate group renders it polymerizable. The precursor was mixed with a cross-linker and their in situ thermal free-radical co-polymerization during nanoimprinting rigidly shaped the patterns, trapped the metal atoms, reduced the surface energy and strengthened the structures, thereby giving a ~100% yield after demolding. The imprinted structures were heat-treated, leading to the loss of organics and their subsequent shrinkage. Amorphous Al2O3 patterns with line-widths as small as 17 nm were obtained. Our process utilizes the advantages of sol-gel and methacrylate routes for imprinting and at the same time alleviates the disadvantages associated with both these methods. With these benefits, the chelating monomer route may be the harbinger of the universal scheme for direct nanoimprinting of metal oxides.

Direct patterning of TiO2 using step-and-flash imprint lithography.

Although step-and-flash imprint lithography, or S-FIL, has brought about tremendous advancement in wafer-scale fabrication of sub-100 nm features of photopolymerizable organic and organo-silicon-based resists, it has not been successful in direct patterning of inorganic materials such as oxides because of the difficulties associated with resist formulation and its dispensing. In this paper, we demonstrate the proof-of-concept S-FIL of titanium dioxide (TiO(2)) carried by an acrylate-based formulation containing an allyl-functionalized titanium complex. The prepolymer formulation contains 48 wt % metal precursor, but it exhibits low enough viscosity (∼5 mPa·s) to be dispensed by an automatic dispensing system, adheres and spreads well on the substrate, is insensitive to pattern density variations, and rapidly polymerizes when exposed to broadband UV radiation to give a yield close to 95%. Five fields, each measuring 1 cm × 1 cm, consisting of 100 nm gratings were successively imprinted. Heat-treatment of the patterned structures at 450 °C resulted in the loss of organics and their subsequent shrinkage without the loss of integrity or aspect ratio and converted them to TiO(2) anatase nanostructures as small as 30 nm wide. With this approach, wafer-scale direct patterning of functional oxides on a sub-100 nm scale using S-FIL can become a distinct possibility.

Direct imprinting of high resolution TiO(2) nanostructures.

We demonstrate a different approach to direct nanoimprint lithography of oxides, in particular TiO(2), using the metal methacrylate route which not only gives very high resolution ( approximately 20 nm) but also provides yields of approximately 100% over areas > 1 cm x 1 cm. TiO(2) was imprinted using a polymerizable liquid 'TiO(2) resin' consisting of a mixture of titanium methacrylate, ethylene glycol dimethacrylate, and azobis-(isobutyronitrile). The resin underwent free radical polymerization when imprinted using a silicon mold at 110 degrees C with pressures as low as 10 bar. Polymerization strengthens the imprinted structures, thereby giving approximately 100% yield after demolding. Heat-treatment of the imprinted structures at 400 degrees C resulted in the loss of organics and their subsequent shrinkage ( approximately 75%) without the loss of integrity or aspect ratio, and converted them to TiO(2) nanostructures as small as approximately 20 nm wide. Furthermore, our method demonstrates that large imprinted areas of sub-100-nm features can be achieved by sub-micron molds which translate into huge cost savings with the added flexibility of direct patterning of urinary as well as multi-component oxides.