Effect of the Polymer Modification on the Cytocompatibility of Human and Rat Cells

Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable, semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H2O and CO2. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells

Polymer carriers modified by plasma and functionalized with Au nanoparticles as substrates for mouse 3T3 fibroblasts

Polymers have been o f ten applied in biology and medicine for construction of tissue replacements. However, the inert surface o f the most polymers is not able to support and control cell adhesion, migration, proliferation, differentiation and other cell functions. Hence, the modification of polymer surface led to achieve appropriate properties. The polymer surface can be modified by plasma discharge by which the polymer surface chemistry and morphology is changed. Plasma treatment leads to creation of radicals, unsaturated bonds and new chemical groups, mainly oxygen containing groups. Oxidized groups increase the wettability of polymers, which supports adsorption of cell adhesion-mediating extracellular matrix (ECM) molecules in appropriate spatial conformation increasing accessibility of specific sites in these molecules by cell adhesion receptors. In addition, other surface properties of polymers are altered by plasma etching which strongly influence cell-material interaction. Radicals and unsaturated chemical bonds which are created by plasma can be utilized for grafting new chemical groups, biomolecules and nanoparticles. The biomolecules grafted on the polymer surface, such as amino acids, RGD-containing oligopeptides (i.e., ligands for integrin receptors), ECM molecules, enazymes, hormones, and also carbon and gold nanoparticles, not only have specific biological effects on cells but also change physical and chemical properties of the polymer surface, and by this way they support its bioactivity. This study is focused on physiochemical properties and biocompatibility of modified polymers. The studied materials were poly(L-lactide) (PLLA) foils, nanofibrous PLLA meshes and polyethylene terephtalate (PTFE) foils. PLLA and PTFE foils were modified in plasma with Ar + ions for time intervals of 50, 10 0 and 300 s with power 8 W, and then grafted with Au nanoparticles. Changes in the surface wettability were determined by reflection goniometry . The presence an d concentration o f Au nanoparticles were examine d by X-ray Photoelectron Spectroscopy (XPS). For the biocompatibility testing, the polymers were seeded by mouse embryonic fibroblasts of the line 3T3, i.e., t he cells of ten utilized as a feeder for keratinocytes. The cell adhesion and growth was evaluated by the number of cells, their morphology and the size of cell adhesion area in the 1st, 3rd and 6th day after seeding. The results indicate that the water drop contact angle increases with the time of exposure to plasma, which means that the vettability decreases. However, the following exposure of plasma-irradiated polymers to a sodium citrate solution (i.e., a storage solution for Au nanoparticles) and grafting with Au nanoparticles decrease the contact angle, i.e., increase the material surface wettability. Our tests of biocompatibility indicate that the modification of the polymer surface in fluences positively the cell behavior. The cells adhered at higher numbers and by a larger cell adhesion area on modified polymers; it was mainly manifested on PTFE

Surface characterization of plasma treated polymers for applications as biocompatible carriers

The objective of this work was to determine surface properties of polymer surfaces after plasma treatment with the aim of further cytocompatibility tests. Examined polymers were poly(ethyleneterephthalate) (PET), high-density polyethylene (HDPE), poly(tetrafluoro-ethylene) (PTFE) and poly(L-lactic acid) (PLLA). Goniometry has shown that the plasma treatment was immediately followed by a sharp decrease of contact angle of the surface. In the course of ageing the contact angle increased due to the reorientation of polar groups into the surface layer of polymer. Ablation of polymer surfaces was observed during the degradation. Decrease of weight of polymer samples was measured by gravimetry. Surface morphology and roughness was studied by atomic force microscopy (AFM). The PLLA samples exhibited saturation of wettability (aged surface) after approximately 100 hours, while the PET and PTFE achieved constant values of contact angle after 336 hours. Irradiation by plasma leads to polymer ablation, the highest mass loss being observed for PLLA. The changes in the surface roughness and morphology were observed, a lamellar structure being induced on PTFE. Selected polymer samples were seeded with VSMC (vascular smooth muscle cells) and the adhesion and proliferation of cells was studied. It was proved that certain combination of input treatment parameters led to improvement of polymer cytocompatibility. The plasma exposure was confirmed to significantly improve the PTFE biocompatibility

Submicron Laser-Induced Dot, Ripple and Wrinkle Structures and Their Applications

Polymers exposed to laser or plasma treatment or modified with different wet methods which enable the introduction of nanoparticles or biologically active species, such as amino-acids, may find many applications both as biocompatible or anti-bacterial materials or on the contrary, can be applied for a decrease in the number of cells on the treated surface which opens application in single cell units. For the experiments, two types of materials were chosen, a representative of non-biodegradable polymers, polyethersulphone (PES) and polyhydroxybutyrate (PHB) as biodegradable material. Exposure of solid substrate to laser well below the ablation threshold can lead to formation of various surface structures. The ripples have a period roughly comparable to the wavelength of the incident laser radiation, and their dimensions depend on many factors, such as chemical composition of the polymer substrate, laser wavelength and the angle of incidence. On the contrary, biopolymers may significantly change their surface roughness and thus influence cell compatibility. The focus was on the surface treatment of PES and PHB by pulse excimer KrF laser with wavelength of 248 nm. The changes of physicochemical properties, surface morphology, surface chemistry and ablation of exposed polymers were studied both for PES and PHB. Several analytical methods involving atomic force microscopy, gravimetry, scanning electron microscopy and others were used for the analysis of the treated surface. It was found that the combination of certain input parameters leads not only to the formation of optimal narrow pattern, but to the combination of a ripple and a wrinkle-like structure, which could be an optimal candidate for cell attachment. The interaction of different types of cells and their interactions with the laser exposed surface were studied. It was found that laser treatment contributes as a major factor for wettability/contact angle change. The combination of optimal laser energy and pulse number was used for the construction of a surface with an anti-cellular response. Due to the simple laser treatment, we were able to prepare a biopolymer surface with higher roughness and thus significantly influence the area of growth of different types of cells (U-2 OS cells)