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The present publication reports the purification effort of two natural bone blocks, that is, an allogeneic bone block (maxgraft®, botiss biomaterials GmbH, Zossen, Germany) and a xenogeneic block (SMARTBONE®, IBI S.A., Mezzovico Vira, Switzerland) in addition to previously published results based on histology. Furthermore, specialized scanning electron microscopy (SEM) and in vitro analyses (XTT, BrdU, LDH) for testing of the cytocompatibility based on ISO 10993-5/-12 have been conducted. The microscopic analyses showed that both bone blocks possess a trabecular structure with a lamellar subarrangement. In the case of the xenogeneic bone block, only minor remnants of collagenous structures were found, while in contrast high amounts of collagen were found associated with the allogeneic bone matrix. Furthermore, only island-like remnants of the polymer coating in case of the xenogeneic bone substitute seemed to be detectable. Finally, no remaining cells or cellular remnants were found in both bone blocks. The in vitro analyses showed that both bone blocks are biocompatible. Altogether, the purification level of both bone blocks seems to be favorable for bone tissue regeneration without the risk for inflammatory responses or graft rejection. Moreover, the analysis of the maxgraft® bone block showed that the underlying purification process allows for preserving not only the calcified bone matrix but also high amounts of the intertrabecular collagen matrix.
Background/Aim: The aim of this study was the development of a new osteoconductivity index to determine the bone healing capacities of bone substitute materials (BSM) on the basis of 3D microcomputed tomographic (μ-CT) data. Materials and Methods: Sinus biopsies were used for the comparative analysis of the integration behavior of two xenogeneic BSM (cerabone® and Bio Oss®). 3D μ-CT and data sets from histomorphometrical measurements based on 2D histological slices were used to measure the bone-material-contact and the tissue distribution within the biopsies. The tissue reactions to both BSM were microscopically analyzed. Results: The 3D and 2D results of the osteoconductivity measurements showed comparable material-bone contacts for both BSM, but the 2D data were significantly lower. The same results were found when tissue distribution was measured in both groups. The histopathological analysis showed comparative tissue reactions in both BSM. Conclusion: Osteoconductivity index is a reliable measurement parameter for determining the healing capacities of BSM. The observed differences between both measurement methods could be assigned to the resolution capacity of μ-CT data that did not allow for a precise interface distinction between both BSM and bone tissue. Histomorphometrical data based on histological slides still allow for a more exact evaluation.
Surface topographies are often discussed as an important parameter influencing basic cell behavior. Whereas most in vitro studies deal with microstructures with sharp edges, smooth, curved microscale topographies might be more relevant concerning in-vivo situations. Addressing the lack of highly defined surfaces with varying curvature, we present a topography chip system with 3D curved features of varying spacing, curvature radii as well as varying overall dimensions of curved surfaces. The CurvChip is produced by low-cost photolithography with thermal reflow, subsequent (repetitive) PDMS molding and hot embossing. The platform facilitates the systematic in-vitro investigation of the impact of substrate curvature on cell types like epithelial, endothelial, smooth muscle cells, or stem cells. Such investigations will not only help to further understand the mechanism of curvature sensation but may also contribute to optimize cell-material interactions in the field of regenerative medicine.
In this study, a novel strategy has been developed for the assembly of polyelectrolyte multilayer (PEM) on CaCO3 templates in acidic pH solutions, where consecutive polyelectrolyte layers (heparin/poly(allylamine hydrochloride) or heparin/chitosan) were deposited on PEM hollow microcapsules established previously on CaCO3 templates. The PEM build-up, hollow capsule characterization and successful encapsulation of fluorescein 5(6)-isothiocyanate (FITC)-Dextran by coprecipitation with CaCO3 are demonstrated. Improvement by the removal of CaCO3 core was achieved while the depositions. In the course of the release profile, high retardation for encapsulated FITC-Dextran was observed. The combined shell capsules system is a significant trait that has potential use in tailoring functional layer-by-layer capsules as intelligent drug delivery vehicles where the preliminary in vitro tests showed the responsiveness on the enzymes.
In this article, liposome-based coatings aiming to control drug release from therapeutic soft contact lenses (SCLs) materials are analyzed. A PHEMA based hydrogel material loaded with levofloxacin is used as model system for this research. The coatings are formed by polyelectrolyte layers containing liposomes of 1,2-dimyristoyl-sn-glycero-3- phosphocholine (DMPC) and DMPC1cholesterol (DMPC1 CHOL). The effect of friction and temperature on the drug release is investigated. The aim of the friction tests is to simulate the blinking of the eyelid in order to verify if the SCLs materials coated with liposomes are able to keep their properties, in particular the drug release ability. It was observed that under the study conditions, friction did not affect significantly the drug release from the liposome coated PHEMA material. In contrast, increasing the temperature of release leads to an increase of the drug diffusion rate through the hydrogel. This phenomenon is recorded both in the control and in the coated samples.
Polyelectrolyte multilayer (PEM) are thin polymeric films produced by alternating adsorption of positively and negatively charged polyelectrolytes (PE) on a substrate. These films are considered drug delivery agents as well as coating material for implants, due to their antibiofouling and biologically benign properties. For these reasons the film mechanical properties as well as response to mechanical stress are important measurement parameters. Especially intriguing is the correlation of the mechanical properties of PEM on macroscopic level with the structure of PEM on molecular level, which is addressed here for the first time. This study investigates PEM from PDADMA/PSS produced by spraying technique with neutron and X-ray reflectometry. Reflectometry technique provides precise information on thickness and density (i.e., electron density or scattering length density, respectively), and, this way, allows to conclude on changes in film composition. Thus, neutron and X-ray reflectometry technique is suitable to investigate the overall and the internal transformations, which PEM films might undergo upon exposure to mechanical load. During uniaxial elongation two regimes of PEM deformation can be observed: An elastic regime at small elongations (below ca. 0.2%), which is characterized by a reversible change of film thickness, and a plastic regime with a permanent change above this limit. Both regimes have in common, that the mechanical load induces an increase of the film thickness, which is accompanied by an uptake of water from the surrounding atmosphere. The strain causes a molecular rearrangement within the PEM-structure of stratified layers, which, even in elastic regime, is permanent, although the thickness change remains reversible.
Polyethylene glycol (PEG) is a widely used modification for drug delivery systems. It reduces undesired interaction with biological components, aggregation of complexes and serves as a hydrophilic linker of ligands for targeted drug delivery. However, PEGylation can also lead to undesired changes in physicochemical characteristics of chitosan/siRNA nanoplexes and hamper gene silencing.
To address this conflicting issue, PEG-chitosan copolymers were synthesized with stepwise increasing degrees of PEG substitution (1.5% to 8.0%). Subsequently formed PEG-chitosan/siRNA nanoplexes were characterized physicochemically and biologically. The results showed that small ratios of chitosan PEGylation did not affect nanoplex stability and density. However, higher PEGylation ratios reduced nanoplex size and charge, as well as cell uptake and final siRNA knockdown efficiency.
Therefore, we recommend fine-tuning of PEGylation ratios to generate PEG-chitosan/siRNA delivery systems with maximum bioactivity. The degree of PEGylation for chitosan/siRNA nanoplexes should be kept low in order to maintain optimal nanoplex efficiency.
Foam has been employed as an improved or enhanced oil recovery method to overcome gravity override and the channeling and fingering of the injected gas, which arises because of the low density and viscosity of the injected fluid combined with the rock heterogeneity. A major challenge, however, is the stability of the generated foam when it contacts the oil. In this study we investigate the feasibility of using inexpensive nanoparticles made of coal fly ash, an abundantly available waste product of coal power plants, as a foam booster. We investigate the viability of reducing the size of fly ash particles to 100−200 nm using high-frequency ultrasonic grinding. We also study the foaminess (foamability), strength, and stability of the foams made with minor concentrations of fly ash nanoparticles and surfactant, both in bulk and porous media. The effect of monovalent and divalent ion concentration on the foaminess of the nanoash suspension combined with very low concentrations of a commercial alpha olefin sulfonate (AOS) surfactant, in the presence and absence of oil, is studied. We observe that bulk foam that contains very small amounts of nanoash particles shows a higher stability in the presence of model oils. Furthermore, experiments in porous media exhibit remarkably stronger foam with mixtures of nanoash and surfactant, such that the amount of produced liquids from the cores significantly increases. For the first time we show that nanoash can be used to stabilize nitrogen foam in the presence of crude oil at high temperature and pressure. In the presence of oil, the nanoash−AOS foam shows a higher stability, although crude oil tends to form stable emulsions in water in the presence of nanoash.
The effect of Hofmeister anions on the surface properties of polyelectrolyte multilayers built from hyaluronan and chitosan by layer-by-layer deposition is studied by ellipsometry and atomic force microscopy. The thickness, roughness and morphology of the resulting coatings were found to depend on the type of the anion. Relationship between the surface properties and the biological response of the polyelectrolyte multilayers is established by assessing the degree of protein (albumin) adsorption.
The interaction between lipid bilayers in water has been intensively studied over the last decades. Osmotic stress was applied to evaluate the forces between two approaching lipid bilayers in aqueous solution. The force–distance relation between lipid mono- or bilayers deposited on mica sheets using a surface force apparatus (SFA) was also measured. Lipid stabilised foam films offer another possibility to study the interactions between lipid monolayers. These films can be prepared comparatively easy with very good reproducibility. Foam films consist usually of two adsorbed surfactant monolayers separated by a layer of the aqueous solution from which the film is created. Their thickness can be conveniently measured using microinterferometric techniques. Studies with foam films deliver valuable information on the interactions between lipid membranes and especially their stability and permeability. Presenting inverse black lipid membrane (BLM) foam films supply information about the properties of the lipid self-organisation in bilayers. The present paper summarises results on microscopic lipid stabilised foam films by measuring their thickness and contact angle. Most of the presented results concern foam films prepared from dispersions of the zwitterionic lipid 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC) and some of its mixtures with the anionic lipid — 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DMPG).
The strength of the long range and short range forces between the lipid layers is discussed. The van der Waals attractive force is calculated. The electrostatic repulsive force is estimated from experiments at different electrolyte concentrations (NaCl, CaCl2) or by modification of the electrostatic double layer surface potential by incorporating charged lipids in the lipid monolayers. The short range interactions are studied and modified by using small carbohydrates (fructose and sucrose), ethanol (EtOH) or dimethylsulfoxide (DMSO). Some results are compared with the structure of lipid monolayers deposited at the liquid/air interface (monolayers spread in Langmuir trough), which are one of most studied biomembrane model system. The comparison between the film thickness and the free energy of film formation is used to estimate the contribution of the different components of the disjoining pressure to the total interaction in the film and their dependence on the composition of the film forming solution.