Drug release from thin films encapsulated by a temperature-responsive hydrogel.

Control over drug delivery may be interestingly achieved by using temperature responsive encapsulants, which change their thickness and mesh size with temperature. The prototype N-isopropylacrylamide hydrogel cross-linked with di(ethylene glycol) divinyl ether p(NIPAAm-co-DEGDVE) swells at low temperature and collapses above the lower critical solution temperature (LCST), ∼29 °C in a buffer. It might be expected that drug release from such encapsulation is always favored below the LCST, due to the larger free volume present in the swollen polymer film. Recent results show contradicting behavior where some cases behave as expected and others release much less when the polymer layer is swollen. In this study, layers of the drugs phenytoin, clotrimazole and indomethacin were drop cast on glass and p(NIPAAM-co-DEGDVE) layers were then synthesized directly on top of these drug layers via initiated chemical vapor deposition (iCVD), a solvent-free and gentle polymerization technique. Dissolution experiments were then performed, in which the drug release through the hindrance of the hydrogel was measured at different pH values. The results show that not only the swelling but also the permeate (drug in this case)-polymer interaction plays an important role in the release.

Crystal alignment of caffeine deposited onto single crystal surfaces via hot-wall epitaxy.

Defined crystal growth is highly demanded for technological applications but also fundamental research. Within this work, the crystal growth of the asymmetric molecule caffeine was studied on single crystalline surfaces of muscovite mica, sodium chloride and potassium chloride. While elongated needle-like crystals grow on muscovite mica and sodium chloride, smaller individual "bird-like" structures were observed on potassium chloride. Depending on the surface type and temperature, the disk-shaped caffeine molecules prefer either an edge-on or flat-on orientation with respect to the surface, but in each case, a defined crystallographic relation between the surface and caffeine crystallites was determined by using the X-ray pole figure technique. On muscovite mica and sodium chloride, needle-like crystallites with edge-on oriented molecules aligned mainly with the unit cell c-axis (which coincides with the long needle axis) along the [1-10]mica, [100]mica, [110]mica and [110]NaCl, [1-10]NaCl directions, respectively. Crystals consisting of flat-on oriented molecules on KCl showed also defined alignments with respect to the substrate, but due to the altered molecule-substrate contact, the b-axis aligned along [110]KCl and [1-10]KCl. Growth at elevated temperatures enabled changes in the crystal growth whereby more defined structures formed on NaCl. On KCl, the bird-like structures remained very similar, while caffeine on the mica surface at elevated temperatures resulted in even additional texture forming with the caffeine molecules now also favoring a flat-on orientation with respect to the surface. The systematic variation of various system parameters demonstrates how sensitive the growth behavior of caffeine on this variety of substrates is.

Wrinkle formation in a polymeric drug coating deposited via initiated chemical vapor deposition.

Polymer encapsulation of drugs is conventionally used as a strategy for controlled delivery and enhanced stability. In this work, a novel encapsulation approach is demonstrated, in which the organic molecule clotrimazole is enclosed into wrinkles of defined sizes. Having defined wrinkles at the drug/encapsulant interface, the contact between the encapsulating polymer and the drug can be improved. In addition, this can also allow for some control on the drug delivery as the available surface area changes with the wrinkle size. For this purpose, thin films of clotrimazole were deposited onto silica substrates and were then encapsulated by crosslinked poly(2-hydroxyethyl methacrylate) (pHEMA) via initiated chemical vapor deposition (iCVD). The thickness and the solid state (crystalline or amorphous) of the clotrimazole layer were varied so that the conditions under which surface wrinkles emerge can be determined. A (critical) clotrimazole thickness of 76.6 nm was found necessary to induce wrinkles, whereby the wrinkle size is directly proportional to the thickness of the amorphous clotrimazole. When the pHEMA was deposited on top of crystalline clotrimazole instead, wrinkling was absent. The wrinkling effect can be understood in terms of elastic mismatch between the relatively rigid pHEMA film and the drug layer. In the case of amorphous clotrimazole, the relatively soft drug layer causes a large mismatch resulting in a sufficient driving force for wrinkle formation. Instead, the increased elastic modulus of crystalline clotrimazole reduces the elastic mismatch between drug and polymer, so that wrinkles do not form.

Thermal stability and molecular ordering of organic semiconductor monolayers: effect of an anchor group.

The thermal stability and molecular order in monolayers of two organic semiconductors, PBI-PA and PBI-alkyl, based on perylene derivatives with an identical molecular structure except for an anchor group for attachment to the substrate in PBI-PA, are reported. In situ X-ray reflectivity measurements are used to follow the stability of these monolayers in terms of order and thickness as temperature is increased. Films have thicknesses corresponding approximately to the length of one molecule; molecules stand upright on the substrate with a defined structure. PBI-PA monolayers have a high degree of order at room temperature and a stable film exists up to 250 °C, but decomposes rapidly above 300 °C. In contrast, stable physisorbed PBI-alkyl monolayers only exist up to 100 °C. Above the bulk melting point at 200 °C no more order exists. The results encourage using anchor groups in monolayers for various applications as it allows enhanced stability at the interface with the substrate.

Particular film formation of phenytoin at silica surfaces.

Given the increasing number of poorly soluble and thus poorly bioavailable active pharmaceutical materials, there is a demand for innovative formulation platforms for such molecules. Thus a focus on enhancing dissolution properties of poorly soluble drugs exists. Within this study, the spin coating of acetone solutions containing 5,5-diphenyl-2,4-imidazolidinedione (phenytoin) in various concentrations is evaluated. The results reveal strong variations of the morphology of deposited phenytoin crystals at silica surfaces. Individual separated particles are obtained on low phenytoin concentrations, and closely packed particular films form when the concentration is increased. As the material is isomorphic, these various morphologies have the same crystalline structure. Dissolution experiments reveal that both the apparent maximum solubility and as the dissolution rate are strongly enhanced compared to bulk powder, suggesting that formulation based on this preparative technique will allow overcoming the low solubility problematic for a variety of drugs.

Crystallographic textures and morphologies of solution cast Ibuprofen composite films at solid surfaces.

The preparation of thin composite layers has promising advantages in a variety of applications like transdermal, buccal, or sublingual patches. Within this model study the impact of the matrix material on the film forming properties of ibuprofen-matrix composite films is investigated. As matrix materials polystyrene, methyl cellulose, or hydroxyl-ethyl cellulose were used. The film properties were either varied by the preparation route, i.e., spin coating or drop casting, or via changes in the relative ratio of the ibuprofen and the matrix material. The resulting films were investigated via X-ray diffraction and atomic force microscope experiments. The results show that preferred (100) textures can be induced via spin coating with respect to the glass surface, while the drop casting results in a powder-like behavior. The morphologies of the films are strongly impacted by the ibuprofen amount rather than the preparation method. A comparison of the various matrix materials in terms of their impact on the dissolution properties show a two times faster zero order release from methyl cellulose matrix compared to a polystyrene matrix. The slowest rate was observed within the hydroxyl ethyl cellulose as the active pharmaceutical ingredients (APIs) release is limited by diffusion through a swollen matrix. The investigation reveals that the ibuprofen crystallization and film formation is only little effected by the selected matrix material than that compared to the dissolution. A similar experimental approach using other matrix materials may therefore allow to find an optimized composite layer useful for a defined application.

Dissolution testing of hardly soluble materials by surface sensitive techniques: clotrimazole from an insoluble matrix.

PURPOSE: The low aqueous solubility of many drugs impedes detailed investigation as the detection limit of standard testing routines is limited. This is further complicated within application relevant thin films typical used in patches or stripes for buccal or topical routes. METHODS: In this work a model system is developed based on spin - casting technique allowing defined clotrimazole and clotrimazole - polystyrene composite films preparation at a solid surface. Various highly sensitive techniques including quarz crystal microbalance (QCM), X-ray reflevtivity (XRR) and X-ray photon spectroscopy (XPS) are used to investigate the drug release over time into an aqueous media. RESULTS: The results reveal a steady drug release for both samples over the course of the experiments but with the release from the composite being significantly slower. In addition the dissolution rate of the clotrimazole sample initially increases up to 30 min after which a decrease is noted. XRR shows that this is a result of surface roughening together with film thickness reduction. The results for the composite show that the release in the composite film is a result of drug diffusion within the matrix and collapsing PS film thickness whereby XPS shows that the amount of clotrimazole at the surface after 800 min immersion is still high. CONCLUSION: It can be stated that the applied techniques allow following low mass drug release in detail which may also be applied to other systems like pellets or surface loaded nano-carriers providing information for processing and application relevant parameters.

Interfacial Engineering of Soft Matter Substrates by Solid-State Polymer Adsorption.

Polymer coating to substrates alters surface chemistry and imparts bulk material functionalities with a minute thickness, even in nanoscale. Specific surface modification of a substate usually requires an active substrate that, e.g., undergoes a chemical reaction with the modifying species. Here, we present a generic method for surface modification, namely, solid-state adsorption, occurring purely by entropic strive. Formed by heating above the melting point or glass transition and subsequent rinsing of the excess polymer, the emerging ultrathin (<10 nm) layers are known in fundamental polymer physics but have never been utilized as building blocks for materials and they have never been explored on soft matter substrates. We show with model surfaces as well as bulk substrates, how solid-state adsorption of common polymers, such as polystyrene and poly(lactic acid), can be applied on soft, cellulose-based substrates. Our study showcases the versatility of solid-state adsorption across various polymer/substrate systems. Specifically, we achieve proof-of-concept hydrophobization on flexible cellulosic substrates, maintaining irreversible and miniscule adsorption yet with nearly 100% coverage without compromising the bulk material properties. The method can be considered generic for all polymers whose Tg and Tm are below those of the to-be-coated adsorbed layer, and whose integrity can withstand the solvent leaching conditions. Its full potential has broad implications for diverse materials systems where surface coatings play an important role, such as packaging, foldable electronics, or membrane technology.

Dynamics of monolayer-island transitions in 2,7-dioctyl-benzothienobenzthiophene thin films.

The order in molecular monolayers is a crucial aspect for their technological application. However, the preparation of defined monolayers by spin-coating is a challenge, since the involved processes are far from thermodynamic equilibrium. In the work reported herein, the dynamic formation of dioctyl-benzothienobenzothiophene monolayers is explored as a function of temperature by using X-ray scattering techniques and atomic force microscopy. Starting with a disordered monolayer after the spin-coating process, post-deposition self-reassembly at room temperature transforms the initially amorphous layer into a well-ordered bilayer structure with a molecular herringbone packing, whereas at elevated temperature the formation of crystalline islands occurs. At the temperature of the liquid-crystalline crystal-smectic transition, rewetting of the surface follows resulting in a complete homogeneous monolayer. By subsequent controlled cooling to room temperature, cooling-rate-dependent kinetics is observed; at rapid cooling, a stable monolayer is preserved at room temperature, whereas slow cooling causes bilayer structures. Increasing the understanding and control of monolayer formation is of high relevance for achieving ordered functional monolayers with defined two-dimensional packing, for future applications in the field of organic electronics.

Interface induced crystal structures of dioctyl-terthiophene thin films.

Temperature dependent structural and morphological investigations on semiconducting dioctyl-terthiophene (DOTT) thin films prepared on silica surfaces reveals the coexistence of surface induce order and distinct crystalline/liquid crystalline bulk polymorphs. X-ray diffraction and scanning force microscopy measurements indicate that at room temperature two polymorphs are present: the surface induced phase grows directly on the silica interface and the bulk phase on top. At elevated temperatures the long-range order gradually decreases, and the crystal G (340 K), smectic F (348 K), and smectic C (360 K) phases are observed. Indexation of diffraction peaks reveals that an up-right standing conformation of DOTT molecules is present within all phases. A temperature stable interfacial layer close to the silica-DOTT interface acts as template for the formation of the different phases. Rapid cooling of the DOTT sample from the smectic C phase to room temperature results in freezing into a metastable crystalline state with an intermediated unit cell between the room temperature crystalline phase and the smectic C phase. The understanding of such interfacial induced phases in thin semiconducting liquid crystal films allows tuning of crystallographic and therefore physical properties within organic thin films.

Ionic liquid nanotribology: mica-silica interactions in ethylammonium nitrate.

Colloid probe atomic force microscopy has been used to study the nanotribological properties of the silica-ethylammonium nitrate (EAN)-mica system. Normal force curve measurements reveal a series of steps at separations that are consistent with the size of an EAN ion pair (0.5 nm) due to displacement of structured solvent layers as the two surfaces are brought together. At closest separations, two steps are measured with widths of 0.3 nm and 0.1 nm, which are too small to be due to an ion pair layer. The 0.3 nm step is attributed to a partial displacement of a silica-bound cation-rich layer, with residual cations being removed in the subsequent 0.1 nm step. Lateral force measurements reveal that the frictional response is dependent on the number of ion pair layers between the surfaces. At low forces, when there is more than a single layer of EAN between silica and mica, the lateral force increases relatively steeply with applied load, and is independent of the sliding speed. At intermediate forces, a single layer of cations in an intercalated bilayer structure is present between the surfaces. The friction coefficient (µ) increases logarithmically with sliding speed consistent with an activated, discontinuous sliding process. At high force, µ is small and once again, independent of sliding velocity. The adsorbed cation layer is bound primarily to mica and compressed by the high normal force. This robust layering with a well-defined sliding plane permits the colloid probe to slide easily over the mica surface.

Impact of sample misalignment on grazing incidence x-ray diffraction patterns and the resulting unit cell determination.

Grazing incidence x-ray diffraction (GIXD) is a frequently used tool for the crystallographic characterization of thin films in terms of polymorph identification and determination of the crystallographic lattice parameters. Even full structure solutions are possible. To obtain highly accurate diffraction patterns, the thin film sample has to be aligned carefully with the center of the goniometer, which allows a defined incidence of the primary x-ray beam relative to the sample surface. This work studies the effect of misalignment of a thin film sample on the acquired diffraction pattern. Three potential types of misalignments are considered: the deviation of the sample surface from the center of the goniometer, an error in the incidence angle of the primary beam, and an inclination of the goniometer rotation axis from the normal of the substrate surface. The consequence of these types of sample misalignments is the shift of diffraction peaks toward specific directions in reciprocal space. Mathematical equations are given that relate the error in positions of Bragg peaks for each type of sample misalignment. Experiments with intentionally misaligned samples confirm the given formulas. In a subsequent step, the errors in the peak positions are translated to systematic errors in the estimation of the unit cell parameters. Depending on the type of misalignment, some alignment errors can be reduced or even corrected; in particular, azimuthal sample rotations prove to be advantageous in these cases. The results in this work improve the quality of GIXD measurements, in general, enabling deeper analysis like the full structure solution from the GIXD pattern on everyday basis.

Compact poly(ethylene oxide) structures adsorbed at the ethylammonium nitrate-silica interface.

The adsorption of poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO) onto silica from ethylammonium nitrate (a protic ionic liquid) has been investigated using colloid probe AFM force curve measurements. Steric repulsive forces were measured for PEO, confirming that PEO can compete with the ethylammonium cation and adsorb onto silica. The range of the repulsion increases with polymer molecular weight (e.g., from 1.4 nm for 0.01 wt % 10 kDa PEO to 40 nm for 0.01 wt % 300 kDa PEO) and with concentration (e.g., from 16 nm at 0.001 wt % to 78 nm at 0.4 wt % for 300 kDa PEO). Fits to the force curve data could not be obtained using standard models for a polymer brush, but excellent fits were obtained using the mushroom model, suggesting the adsorbed polymer films are compressed and relatively poorly solvated. No evidence for adsorption of 3.5 kDa PPO could be detected up to its solubility limit.

Interactions of adsorbed poly(ethylene oxide) mushrooms with a bare silica-ionic liquid interface.

The interaction of adsorbed poly(ethylene oxide) (PEO) mushrooms with clean silica-ethylammonium nitrate (EAN, a protic ionic liquid) interfaces is investigated using atomic force microscopy (AFM). 10 kDa, 35 kDa and 100 kDa PEO was used to prepare polymer layers ex situ by drop casting from 0.01 wt% EAN solutions. AFM tapping mode measurements of dried, solvent free PEO layers revealed oblate structures, which increase in size with molecular weight. Colloid probe force curve measurements of these surfaces re-solvated with EAN suggest PEO adopts a mushroom morphology, with the interaction range (layer thickness) increasing with molecular weight. Attractive forces on approach and single strand stretching forces on retraction show PEO has a strong affinity for the silica-EAN interface. The single polymer strand stretching forces follow the freely jointed chain model under good solvent conditions. Contour lengths close to the theoretical limits of 120 nm for the 10 kDa, 290 nm for the 35 kDa and 1240 nm for the 100 kDa PEO samples are observed, while fitted Kuhn lengths are small, at 0.14 nm.

Fabrication, characterization and cytocompatibility assessment of gelatin nanofibers coated with a polymer thin film by initiated chemical vapor deposition.

The presence of various functional groups in the structure of gelatin nanofibers (GNFs) has made it a suitable candidate for biomedical applications, yet its fast dissolution in aqueous media has been a real challenge for years. In the present work, we propose an efficient procedure to improve the durability of the GNFs. The electrospun GNFs were coated with poly(ethylene glycol dimethacrylate) (pEGDMA) using initiated chemical vapor deposition (iCVD) as a completely dry polymerization method. Morphological and chemical analysis revealed that an ultrathin layer formed around nanofibers (iCVD-GNFs) which has covalently bonded to gelatin chains. Against the instant dissolution of GNFs, the in vitro biodegradability test showed the iCVD-GNFs, to a large extent, preserve their morphology after 14 days of immersion and did not lose its integrity even after 31 days. In vitro cell culture studies, also, revealed cytocompatibility of the iCVD-GNFs for human fibroblast cells (hFC), as well as higher cell proliferation on the iCVD-GNFs compared to control made from tissue culture plate (TCP). Furthermore, contact angle measurements indicated that the hydrophilic GNFs became hydrophobic after the iCVD, yet FE-SEM images of cell-seeded iCVD-GNFs showed satisfactory cell adhesion. Taken together, the proposed method paves a promising way for the production of water-resistant GNFs utilized in biomedical applications; for instance, tissue engineering scaffolds and wound dressings.

Wrinkling of an Enteric Coating Induced by Vapor-Deposited Stimuli-Responsive Hydrogel Thin Films.

In this contribution, we report on the thin-film synthesis of a thermoresponsive polymer onto another polymer used as an enteric coating in drug applications. In particular, we deposit cross-linked poly(N-vinylcaprolactam) (pNVCL) thin films by initiated chemical vapor deposition (iCVD) onto spin-coated Eudragit (EUD) layers. Already upon iCVD synthesis, the layered structure starts to form wrinkles at a minimum iCVD thickness of 30 nm. By changing the EUD layer thickness and the amount of cross-linking used during iCVD, the morphology of the wrinkles is demonstrated to be readily tunable. Laterally, the double-layer structures vary in morphology from being ultrasmooth to exhibiting up to a 3.5 µm wrinkle wavelength. The surface roughness and, thus, the wrinkles' height can be tailored from below 1 nm up to 100 nm. From the resulting wavelength of wrinkles, an estimation of the elastic modulus of pNVCL proves its tunability over a wide range of values thanks to the iCVD process. This study elucidates an uncomplicated way to tune the wrinkles' morphology and, thus, the surface area of a system that can be employed in drug delivery applications. Hence, an enteric coating of EUD together with an iCVD-synthesized thermoresponsive thin film is proposed as a promising composite encapsulation layer to outperform established systems in terms of tunability of the response to multiple external stimuli.

A Protocol To Characterize Peptide-Based Drug Delivery Systems for miRNAs.

Micro RNA (miRNA)-based medicines have attracted attention as new therapeutic strategies to treat genetic diseases and metabolic and immunological disorders. MiRNAs have emerged as key mediators of metabolic processes fulfilling regulatory functions in maintaining physiological conditions, while altered miRNA expression profiles are often associated with genetic diseases. However, naked miRNAs exhibit poor enzymatic stability, biomembrane permeation, and cellular uptake. To overcome these limitations, the development of appropriate drug delivery systems (DDS) is necessary. Herein, a DDS is characterized being assembled from miRNA-27a (negative regulator in fat metabolism) and the amphipathic N-TER peptide. Dynamic light scattering (DLS), electrophoretic light scattering, and atomic force microscopy (AFM) are used to investigate physicochemical properties (i.e., size, shape, and charge) of the DDS. Although surface charges should provide decent stabilization, the AFM results confirm a state of agglomeration, which is also suggested by DLS. Furthermore, AFM studies reveal adhesion on hydrophilic as well as hydrophobic substrates, which is related to the amphipathic properties of the N-TER peptide. Physicochemical properties of DDS are important parameters, which have an impact on cell internalization/uptake and have to be taken into account for in vitro studies to develop a successful peptide-based DDS for miRNA replacement therapy in metabolic diseases, such as obesity and others.

Interlink between Tunable Material Properties and Thermoresponsiveness of Cross-Linked Poly(N-vinylcaprolactam) Thin Films Deposited by Initiated Chemical Vapor Deposition.

In this contribution, we report on the thin-film synthesis of a novel thermoresponsive polymer, namely, poly(N-vinylcaprolactam) cross-linked by di(ethylene glycol)divinyl ether [p(NVCL-co-DEGDVE)] by initiated chemical vapor deposition (iCVD). Its transition between swollen and shrunken states in film thickness and the corresponding lower critical solution temperature (LCST) was investigated by spectroscopic ellipsometry in water. Water contact angle measurements and nano-indentation experiments reveal that the transition is accompanied by a change in wettability and elastic modulus. The amount of cross-linking was used to tune the thermoresponsive behavior of the thin films, resulting in higher swelling and LCST, increased surface rearrangement, and lower stiffness for less cross-linked polymers. For the first time, the filament temperature during iCVD synthesis was used to vary the chain length of the resulting polymeric systems and, thus, the position of their thermoresponsive transition. With that, swelling of up to 250% compared to the dry thickness and transition temperatures ranging from 16 to 40 °C could be achieved.

Biological Activity Of miRNA-27a Using Peptide-based Drug Delivery Systems.

BACKGROUND: Endogenously expressed microRNAs (miRNAs) have attracted attention as important regulators in post-transcriptionally controlling gene expression of various physiological processes. As miRNA dysregulation is often associated with various disease patterns, such as obesity, miRNA-27a might therefore be a promising candidate for miRNA mimic replacement therapy by inhibiting adipogenic marker genes. However, application of naked nucleic acids faces some limitations concerning poor enzymatic stability, bio-membrane permeation and cellular uptake. To overcome these obstacles, the development of appropriate drug delivery systems (DDS) for miRNAs is of paramount importance. METHODS: In this work, a triple combination of atomic force microscopy (AFM), brightfield (BF) and fluorescence microscopy was used to trace the cellular adhesion of N-TER peptide-nucleic acid complexes followed by time-dependent uptake studies using confocal laser scanning microscopy (cLSM). To reveal the biological effect of miRNA-27a on adipocyte development after transfection treatment, Oil-Red-O (ORO)- staining was performed to estimate the degree of in lipid droplets accumulated ORO in mature adipocytes by using light microscopy images as well as absorbance measurements. RESULTS: The present findings demonstrated that amphipathic N-TER peptides represent a suitable DDS for miRNAs by promoting non-covalent complexation through electrostatic interactions between both components as well as cellular adhesion of the N-TER peptide - nucleic acid complexes followed by uptake across cell membranes and intracellular release of miRNAs. The anti-adipogenic effect of miRNA-27a in 3T3-L1 cells could be detected in mature adipocytes by reduced lipid droplet formation. CONCLUSION: The present DDS assembled from amphipathic N-TER peptides and miRNAs is capable of inducing the anti-adipogenic effect of miRNA-27a by reducing lipid droplet accumulation in mature adipocytes. With respect to miRNA mimic replacement therapies, this approach might provide new therapeutic strategies to prevent or treat obesity and obesity-related disorders.

Surface Induced Phenytoin Polymorph. 2. Structure Validation by Comparing Experimental and Density Functional Theory Raman Spectra.

A method for structure solution in thin films that combines grazing incidence X-ray diffraction data analysis and crystal structure prediction was presented in a recent work (Braun et al. Cryst. Growth Des.2019, DOI: 10.1021/acs.cgd.9b00857). Applied to phenytoin form II, which is only detected in films, the approach gave a very reasonable, but not fully confirmed, candidate structure with Z = 4 and Z' = 2. In the present work, we demonstrate how, by calculating and measuring the crystal Raman spectrum in the low wavenumber energy region with the aim of validating the candidate structure, this can be further refined. In fact, we find it to correspond to a saddle point of the energy landscape of the system, from which a minimum of lower symmetry may be reached. With the new structure, with Z = 4 and Z' = 2, we finally obtain an excellent agreement between experimental and calculated Raman spectra.