Alexey Kondyurin

Substrate elasticity provides mechanical signals for the expansion of hemopoietic stem and progenitor cells.

Surprisingly little is known about the effects of the physical microenvironment on hemopoietic stem and progenitor cells. To explore the physical effects of matrix elasticity on well-characterized primitive hemopoietic cells, we made use of a uniquely elastic biomaterial, tropoelastin. Culturing mouse or human hemopoietic cells on a tropoelastin substrate led to a two- to threefold expansion of undifferentiated cells, including progenitors and mouse stem cells. Treatment with cytokines in the presence of tropoelastin had an additive effect on this expansion. These biological effects required substrate elasticity, as neither truncated nor cross-linked tropoelastin reproduced the phenomenon, and inhibition of mechanotransduction abrogated the effects. Our data suggest that substrate elasticity and tensegrity are important mechanisms influencing hemopoietic stem and progenitor cell subsets and could be exploited to facilitate cell culture.

Free radical functionalization of surfaces to prevent adverse responses to biomedical devices

Immobilizing a protein, that is fully compatible with the patient, on the surface of a biomedical device should make it possible to avoid adverse responses such as inflammation, rejection, or excessive fibrosis. A surface that strongly binds and does not denature the compatible protein is required. Hydrophilic surfaces do not induce denaturation of immobilized protein but exhibit a low binding affinity for protein. Here, we describe an energetic ion-assisted plasma process that can make any surface hydrophilic and at the same time enable it to covalently immobilize functional biological molecules. We show that the modification creates free radicals that migrate to the surface from a reservoir beneath. When they reach the surface, the radicals form covalent bonds with biomolecules. The kinetics and number densities of protein molecules in solution and free radicals in the reservoir control the time required to form a full protein monolayer that is covalently bound. The shelf life of the covalent binding capability is governed by the initial density of free radicals and the depth of the reservoir. We show that the high reactivity of the radicals renders the binding universal across all biological macromolecules. Because the free radical reservoir can be created on any solid material, this approach can be used in medical applications ranging from cardiovascular stents to heart-lung machines.

A Comparison of Covalent Immobilization and Physical Adsorption of a Cellulase Enzyme Mixture

This paper reports the first use of a linker-free covalent approach for immobilizing an enzyme mixture. Adsorption from a mixture is difficult to control due to varying kinetics of adsorption, variations in the degree of unfolding and competitive binding effects. We show that surface activation by plasma immersion ion implantation (PIII) produces a mildly hydrophilic surface that covalently couples to protein molecules and avoids these issues, allowing the attachment of a uniform monolayer from a cellulase enzyme mixture. Atomic force microscopy (AFM) showed that the surface layer of the physically adsorbed cellulase layer on the mildly hydrophobic surface (without PIII) consisted of aggregated enzymes that changed conformation with incubation time. The evolution observed is consistent with the existence of transient complexes previously postulated to explain the long time constants for competitive displacement effects in adsorption from enzyme mixtures. AFM indicated that the covalently coupled bound layer to the PIII-treated surface consisted of a stable monolayer without enzyme aggregates, and became a double layer at longer incubation times. Light scattering analysis showed no indication of aggregates in the solution at room temperature, which indicates that the surface without PIII-treatment induced enzyme aggregation. A model for the attachment process of a protein mixture that includes the adsorption kinetics for both surfaces is presented.

Attachment of horseradish peroxidase to polytetrafluorethylene (teflon) after plasma immersion ion implantation

The aim of this work was to investigate the potential of polytetrafluorethylene (PTFE) as a surface for biologically active protein attachment. A plasma immersion ion implantation (PIII) treatment was applied to PTFE to produce an activated surface for the functional attachment of the enzyme, horseradish peroxidase (HRP). Fourier transform infrared-attenuated total reflectance spectra show oxidation and carbonization of the surface layer as a function of ion fluence. The PIII treatment increases by threefold the amount of attached HRP and the activity of HRP on the modified surface is about seven times higher than that on an untreated PTFE surface. This result indicates that the PIII surface modification improves both the polymers protein binding capacity and its ability to retain the protein in a bioactive state.

Surface plasma modification and tropoelastin coating of a polyurethane co-polymer for enhanced cell attachment and reduced thrombogenicity.

Polymers currently utilized for dermal and vascular applications possess sub-optimal biocompatibility which reduces their efficacy. Improving the cell-binding and blood-contacting properties of these polymers would substantially improve their clinical utility. Tropoelastin is a highly extensible extracellular matrix protein with beneficial cell interactive and low thrombogenic properties. We transferred these benefits to the polyurethane block copolymer Elast-Eon E2A through a specific combination of surface plasma modifications and coating with human tropoelastin. The cell-binding activity of bound tropoelastin was modulated by ion implantation of the underlying polymer, and correlated with surface hydrophobicity, carbon and oxygen content. This combined treatment enhanced human dermal fibroblast (HDF) and human umbilical vein endothelial cell (HUVEC) attachment, cytoskeletal assembly and viability, combined with elevated PECAM-1 staining of HUVEC cell junctions. The thrombogenicity of the polymer was ameliorated by tropoelastin coating. We propose that a combination of metered plasma treatment and tropoelastin coating of Elast-Eon can serve to improve the biological performance of implantable devices such as vascular conduits.

Oxygen incorporation in Ti2AlC thin films

Thin films of Ti2AlC MAX phase have been deposited using a multiple cathode pulsed cathodic arc. Evidence for substantial oxygen incorporation in the MAX phase is presented, likely originating from residual gas present in the vacuum chamber during deposition. The characteristic MAX phase crystal structure is maintained, in agreement with ab initio calculations, supporting substitutional O in C lattice positions. On the basis of these results, we propose the existence of a MAX phase-like material with material properties tuned by the incorporation of oxygen. Additionally, possible unintentional O incorporation in previously reported MAX phase materials is suggested.

Plasma-based biofunctionalization of vascular implants

Polymeric and metallic materials are used extensively in permanently implanted cardiovascular devices and devices that make temporary but often prolonged contact with body fluids and tissues. Foreign body responses are typically triggered by host interactions at the implant surface, making surface modifications to increase biointegration desirable. Plasma-based treatments are extensively used to modify diverse substrates; modulating surface chemistry, wettability and surface roughness, as well as facilitating covalent biomolecule binding. Each aspect impacts on facets of vascular compatibility including endothelialization and blood contact. These modifications can be readily applied to polymers such as Dacron and expanded polytetrafluoroethylene, which are widely used in bypass grafting and the metallic substrates of stents, valves and pacemaker components. Plasma modification of metals is more challenging given the need for coating deposition in addition to surface activation, adding the necessity for robust interface adhesion. This review examines the evolving plasma treatment technology facilitating the biofunctionalization of polymeric and metallic implantable cardiovascular materials.

Ion beam modification of polyethylene and adhesion to epoxy adhesive

Abstract Change of surface structure, interface of adhesion joint and adhesion of polyethylene after N+ ion beam treatment were studied. Increase of adhesion strength of polyethylene and epoxy adhesive after ion beam treatment is observed. Ion beam treatment can be used for adhesion ability increasing of polyethylene.

Biospectroscopy of Nanodiamond-Induced Alterations in Conformation of Intra- and Extracellular Proteins: A Nanoscale IR Study.

The toxicity of nanomaterials raises major concerns because of the impact that nanomaterials may have on health, which remains poorly understood. We need to explore the fate of individual nanoparticles in cells at nano and molecular levels to establish their safety. Conformational changes in secondary protein structures are one of the main indicators of impaired biological function, and hence, the ability to identify these changes at a nanoscale level offers unique insights into the nanotoxicity of materials. Here, we used nanoscale infrared spectroscopy and demonstrated for the first time that nanodiamond-induced alterations in both extra- and intracellular secondary protein structures lead to the formation of antiparallel β-sheet, β-turns, intermolecular β-sheet, and aggregation of proteins. These conformational changes of the protein structure may result in the loss of functionality of proteins and in turn lead to adverse effects.

Pulse and continuous ion beam treatment of polyethylene

Polyethylene (PE) films were treated by a nitrogen ion beam with an energy of 20 keV at a quasicontinuous regime with low current density and at a pulse-periodical regime with high current density. IR ATR spectra and UV spectra showed significant differences in structural changes of PE treated by pulse and continuous treatment at the same average current density. The changes in the molecular structure that are induced by ion beams, i.e. the appearance of aromatic cycles, unsaturated bonds and carbonyl groups in PE, have a similar qualitative character for all types of ion beam regimes. However, the same degree of structural changes is achieved at lower dose in the pulse regime than in the continuous regime. At equal treatment parameters (ion energy, dose treatment, average current density) the pulse regime leads to a higher concentration of unsaturated and oxygen-containing groups then the continuous regime. This effect at the pulse regime can be explained by the high current density in the single pulse, at which a high local temperature is generated in the ion track field of the polymer. Probably this leads to a wave of internal stress, and these effects additionally stimulate structural changes in the polymer at pulsed ion beam treatment.