Extracellular matrix-inspired surface coatings functionalized with dexamethasone-loaded liposomes to induce osteo- and chondrogenic differentiation of multipotent stem cells.

Biomimetic surface coatings can be combined with conventional implants to mimic the extracellular matrix (ECM) of the surrounding tissue to make them more biocompatible. Layer-by-layer technique (LbL) can be used for making surface coatings by alternating adsorption of polyanions and polycations from aqueous solutions without need of chemical reactions. Here, polyelectrolyte multilayer (PEM) systems is made of hyaluronic acid (HA) as polyanion and Collagen I (Col) as polycation to mimic the ECM of connective tissue. The PEM are combined with dexamethasone (Dex)-loaded liposomes to achieve a local delivery and protection of this drug for stimulation of osteo- and chondrogenic differentiation of multipotent stem cells. The liposomes possess a positive surface charge that is required for immobilization on the PEM. The surface properties of PEM system show a positive zeta potential after liposome adsorption and a decrease in wettability, both promoting cell adhesion and spreading of C3H10T1/2 multipotent embryonic mouse fibroblasts. Differentiation of C3H10T1/2 was more prominent on the PEM system with embedded Dex-loaded liposomes compared to the basal PEM system and the use of free Dex-loaded liposomes in the supernatant. This was evident by immunohistochemical staining and an upregulation of the expression of genes, which play a key role in osteogenesis (RunX2, ALP, Osteocalcin (OCN)) and chondrogenesis (Sox9, aggrecan (ACAN), collagen type II), determined by quantitative Real-time polymerase chain reaction (qRT-PCR) after 21 days. These findings indicate that the designed liposome-loaded PEM system have high potential for use as drug delivery systems for implant coatings that can induce bone and cartilage differentiation needed for example in osteochondral implants.

Investigation of Integrated Reactive Multilayer Systems for Bonding in Microsystem Technology.

For the integration of a reactive multilayer system (iRMS) with a high exothermic reaction enthalpy as a heat source on silicon wafers for low-temperature bonding in the 3D integration and packaging of microsystems, two main conflicting issues should be overcome: heat accumulation arising from the layer interface pre-intermixing, which causes spontaneous self-ignition during the deposition of the system layers, and conductive heat loss through the substrate, which leads to reaction propagation quenching. In this work, using electron beam evaporation, we investigated the growth of a high exothermic metallic Pd/Al reactive multilayer system (RMS) on different Si-wafer substrates with different thermal conduction, specifically a bare Si-wafer, a RuOx or PdOx layer buffering Si-wafer, and a SiO2-coated Si-wafer. With the exception of the bare silicon wafer, the RMS grown on all other coated wafers underwent systematic spontaneous self-ignition surging during the deposition process once it reached a thickness of around 1 µm. This issue was surmounted by investigating a solution based on tuning the output energy by stacking alternating sections of metallic reactive multilayer Pd/Al and Ni/Al systems that have a high and medium enthalpy of exothermic reactions, respectively. This heterostructure with a bilayer thickness of 100 nm was successfully grown on a SiO2-coated Si-wafer to a total thickness of 3 µm without any spontaneous upsurge of self-ignition; it could be electrically ignited at room temperature, enabling a self-sustained propagating exothermic reaction along the reactive patterned track without undergoing quenching. The results of this study will promote the growth of reactive multilayer systems by electron beam evaporation processing and their potential integration as local heat sources on Si-wafer substrates for bonding applications in microelectronics and microsystems technology.

Engineering osteogenic microenvironments by combination of multilayers from collagen type I and chondroitin sulfate with novel cationic liposomes.

Cationic liposomes composed of a novel lipid (N-{6-amino-1-[N-(9Z) -octadec9-enylamino] -1-oxohexan-(2S) -2-yl} -N'- {2- [N, N-bis(2-aminoethyl) amino] ethyl} -2-hexadecylpropandiamide) (OO4) and dioleoylphosphatidylethanolamine (DOPE) possess high amounts of amino groups and are promising systems for lipofection. Moreover, these cationic liposomes can also be used as a polycationic entity in multilayer formation using layer-by-layer technique (LbL), which is a method to fabricate surface coatings by alternating adsorption of polyanions and polycations. Since liposomes are suitable for endocytosis by or fusion with cells, controlled release of their cargo on site is possible. Here, a polyelectrolyte multilayer (PEM) system was designed of chondroitin sulfate (CS) and collagen type I (Col I) by LbL technique with OO4/DOPE liposomes embedded in the terminal layers to create an osteogenic microenvironment. Both, the composition of PEM and cargo of the liposomes were used to promote osteogenic differentiation of C2C12 myoblasts as in vitro model. The internalization of cargo-loaded liposomes from the PEM into C2C12 cells was studied using lipophilic (Rhodamine-DOPE conjugate) and hydrophilic (Texas Red-labeled dextran) model compounds. Besides, the use of Col I and CS should mimic the extracellular matrix of bone for future applications such as bone replacement therapies. Physicochemical studies of PEM were done to characterize the layer growth, thickness, and topography. The adhesion of myoblast cells was also evaluated whereby the benefit of a cover layer of CS and finally Col I above the liposome layer was demonstrated. As proof of concept, OO4/DOPE liposomes were loaded with dexamethasone, a compound that can induce osteogenic differentiation. A successful induction of osteogenic differentiation of C2C12 cells with the novel designed liposome-loaded PEM system was shown. These findings indicate that designed OH4/DOPE loaded PEMs have a high potential to be used as drug delivery or transfection system for implant coating in the field of bone regeneration and other applications.

The Effect of Heat Treatment on the Microstructure and Mechanical Properties of 2D Nanostructured Au/NiFe System.

Nanostructured NiFe film was obtained on silicon with a thin gold sublayer via pulsed electrodeposition and annealed at a temperature from 100 to 400 °C in order to study the effect of heat treatment on the surface microstructure and mechanical properties. High-resolution atomic force microscopy made it possible to trace stepwise evolving microstructure under the influence of heat treatment. It was found that NiFe film grains undergo coalescence twice-at ~100 and ~300 °C-in the process of a gradual increase in grain size. The mechanical properties of the Au/NiFe nanostructured system have been investigated by nanoindentation at two various indentation depths, 10 and 50 nm. The results showed the opposite effect of heat treatment on the mechanical properties in the near-surface layer and in the material volume. Surface homogenization in combination with oxidation activation leads to abnormal strengthening and hardening-up of the near-surface layer. At the same time, a nonlinear decrease in hardness and Young's modulus with increasing temperature of heat treatment characterizes the internal volume of nanostructured NiFe. An explanation of this phenomenon was found in the complex effect of changing the ratio of grain volume/grain boundaries and increasing the concentration of thermally activated diffuse gold atoms from the sublayer to the NiFe film.

In vitro evaluation of drug release and antibacterial activity of a silver-loaded wound dressing coated with a multilayer system.

The goal of the study was to elaborate an antibacterial silver wound dressing covered by a protective coating that would prevent silver diffusion toward skin without losing its biocide properties. Therefore, non woven polyethyleneterephtalate (PET) textiles were pre-treated by two types of polysaccharides - chitosan and cyclodextrin - both crosslinked with citric acid by a pad/dry/cure process. Both types of resulting thermofixed textiles carrying the citrate crosslinks were then impregnated in silver solution followed by a thermal treatment and were finally coated by Layer-by-Layer (L-b-L) deposition of a polyelectrolyte multilayer (PEM) film consisting of anionic water-soluble poly-cyclodextrin and cationic chitosan. The influence of the process parameters was investigated in terms of silver adsorption capacity, PEM system build-up, silver kinetics of release and antibacterial activity. We demonstrate i) the utility of the intermediate thermal treatment step in the reduction of silver leakage in the polyelectrolyte solutions used in the L-b-L process, ii) that silver adsorption on the preliminary thermofixed layers did not affect the PEM system build-up, iii) the slowing down of silver release kinetic thanks to the PEM coating, iv) the preservation of the antibacterial activity despite the PEM coating.

THz Pulse Detection by Multilayered GeTe/Sb2Te3.

We proposed and demonstrated terahertz (THz) pulse detection by means of multilayered GeTe/Sb2Te3 phase-change memory materials that are also known as a multilayer topological insulator-normal insulator (MTN) system. THz time-domain spectroscopy measurement was performed for MTN films with different multilayer repetitions as well as a conventional as-grown Ge-Te-Sb (GST) alloy film. It was found that MTNs absorb THz waves and that the absorption coefficient depends on the number of layers, while the as-grown GST alloy film was almost transparent for THz waves. Simple MTN-based THz detection devices were fabricated, and the THz-induced change in the current signal was measured when a DC bias voltage was applied between the electrodes. We confirmed that irradiation of THz pulse causes a decrease in the resistance of the MTNs. This result indicates that our devices are capable of THz detection.

Cationic ß-cyclodextrin polymer applied to a dual cyclodextrin polyelectrolyte multilayer system.

A polyelectrolyte multilayer film (PEM) based on cationic and anionic ß-cyclodextrin polyelectrolytes was coated onto a textile substrate for future drug delivery purposes. We firstly synthesized a novel cationic ß-cyclodextrin polymer (polyEPG-CD) by crosslinking ß-cyclodextrin (ßCD) with epichlorohydrin (EP) under basic conditions, in the presence of glycidyltrimetrylammonium chloride (GTMAC) as cationizing group. The influence of preparation conditions has been investigated in order to preferably obtain a water soluble fraction whose charge density and molecular weights were optimal for the layer-by-layer (LbL) deposition process. The different cationic cyclodextrin polymers obtained were characterized by FTIR, NMR, colloidal titration, conductimetry, thermogravimetric analysis and size exclusion chromatography. Besides, the counterpart polyelectrolyte was a ß-cyclodextrin polymer crosslinked with citric acid, polyCTR-CD, whose synthesis and characterization have been previously reported. Finally we realized the Layer by Layer (LbL) build-up of the PEM coating onto the textile support, using the dip coating method, by alternatively soaking it in cationic polyEPG-CD and anionic polyCTR-CD solutions. This multilayer self-assembly was monitored by SEM, gravimetry and OWLS in function of both polyelectrolytes concentrations and ratios. Solutions parameters such as pH, ionic strenght were also discussed.