Improved Adhesion, Growth and Maturation of Vascular Smooth Muscle Cells on Polyethylene Grafted with Bioactive Molecules and Carbon Particles

High-density polyethylene (PE) foils were modified by an Ar+ plasma discharge and subsequent grafting with biomolecules, namely glycine (Gly), polyethylene glycol (PEG), bovine serum albumin (BSA), colloidal carbon particles (C) or BSA and C (BSA + C). As revealed by atomic force microscopy (AFM), goniometry and Rutherford Backscattering Spectroscopy (RBS), the surface chemical structure and surface morphology of PE changed dramatically after plasma treatment. The contact angle decreased for the samples treated by plasma, mainly in relation to the formation of oxygen structures during plasma irradiation. A further decrease in the contact angle was obvious after glycine and PEG grafting. The increase in oxygen concentration after glycine and PEG grafting proved that the two molecules were chemically linked to the plasma-activated surface. Plasma treatment led to ablation of the PE surface layer, thus the surface morphology was changed and the surface roughness was increased. The materials were then seeded with vascular smooth muscle cells (VSMC) derived from rat aorta and incubated in a DMEM medium with fetal bovine serum. Generally, the cells adhered and grew better on modified rather than on unmodified PE samples. Immunofluorescence showed that focal adhesion plaques containing talin, vinculin and paxillin were most apparent in cells on PE grafted with PEG or BSA + C, and the fibres containing α-actin, β-actin or SM1 and SM2 myosins were thicker, more numerous and more brightly stained in the cells on all modified PE samples than on pristine PE. An enzyme-linked immunosorbent assay (ELISA) revealed increased concentrations of focal adhesion proteins talin and vinculin and also a cytoskeletal protein β-actin in cells on PE modified with BSA + C. A contractile protein α-actin was increased in cells on PE grafted with PEG or Gly. These results showed that PE activated with plasma and subsequently grafted with bioactive molecules and colloidal C particles, especially with PEG and BSA + C, promotes the adhesion, proliferation and phenotypic maturation of VSMC

Wettability and Other Surface Properties of Modified Polymers

Surface wettability is one of the crucial characteristics for determining of a material’s use in specific application. Determination of wettability is based on the measurement of the material surface contact angle. Contact angle is the main parameter that characterizes the drop shape on the solid surface and is also one of the directly measurable properties of the phase interface. In this chapter, the wettability and its related properties of pristine and modified polymer foils will be described. The wettability depends on surface roughness and chemical composition. Changes of these parameters can adjust the values of contact angle and, therefore, wettability. In the case of pristine polymer materials, their wettability is unsuitable for a wide range of applications (such as tissue engineering, printing, and coating). Polymer surfaces can easily be modified by, e.g., plasma discharge, whereas the bulk properties remain unchanged. This modification leads to oxidation of the treated layer and creation of new chemical groups that mainly contain oxygen. Immediately after plasma treatment, the values of the contact angles of the modified polymer significantly decrease. In the case of a specific polymer, the strongly hydrophilic surface is created and leads to total spreading of the water drop. Wettability is strongly dependent on time from modification

Substrate Effects of Noble Metal Nanostructures Prepared by Sputtering

Cathode sputtering is a well-established technique for preparation of metal nanostructures. However, the substrate properties are very important in this process. On glass substrates, there is a difficulty with poor adhesion of the metal layers, but thanks to this, metal nanostructures can be produced using solid state dewetting process. Thin metal films on polymer substrates are strongly influenced by the surface properties of the polymers, which originate in the method of their preparation. A recent focus is direct sputtering of metal nanoparticles (NPs) into liquid substrates and their characterizations and applications. Polyethylene glycol (PEG) is one of the most commonly used liquid, which provides “stealth” character to nanostructures. Recent results in this area are reviewed in this chapter. PEGylated NPs could find application in drug delivery systems, therapy, imaging, biosensing, and tissue regeneration

Surface modification of starch based biomaterials by oxygen plasma or UV-irradiation

Radiation is widely used in biomaterials science for surface modification and sterilization. Herein, we describe the use of plasma and UV-irradiation to improve the biocompatibility of different starch-based blends in terms of cell adhesion and proliferation. Physical and chemical changes, introduced by the used methods, were evaluated by complementary techniques for surface analysis such as scanning electron microscopy, atomic force microscopy, contact angle analysis and X-ray photoelectron spectroscopy. The effect of the changed surface properties on the adhesion of osteoblast-like cells was studied by a direct contact assay. Generally, both treatments resulted in higher number of cells adhered to the modified surfaces. The importance of the improved biocompatibility resulting from the irradiation methods is further supported by the knowledge that both UV and plasma treatments can be used as cost-effective methods for sterilization of biomedical materials and devices.I. P. thanks the FCT for providing her a postdoctoral scholarship (SFRH/BPD/8491/2002). This work was partially supported by FCT, through funds from the POCTI and/or FEDER programs, The European Union funded STREP Project HIPPOCRATES (NNM-3-CT-2003-505758) and the European NoE EXPERTISSUES (NMP3-CT-2004-500283)

Hydroxyapatite reinforcement of different starch-based polymers affects osteoblast-like cells adhesion/spreading and proliferation

The aim of this study was to determine which, from a range of the starch-based biomaterials, would be more suitable to be used in orthopaedic applications. This included blends of corn starch and ethylene vinyl alcohol (SEVA-C), corn starch and cellulose acetate (SCA), corn starch and polycaprolactone (SPCL) and its composites with increasing percentages of hydroxyapatite (HA). Osteoblast-like cells (SaOs-2) were cultured in direct contact with the polymers and composites and the effect of the incorporation and of increasing percentages of the ceramic in osteoblast adhesion/proliferation was assessed. In the evaluation of cell adhesion and proliferation rate, two variables were considered; cells adhered to the bottom of the tissue culture polystyrene wells (TCPS) and cells adhered to the surface of the materials, in order to distinguish, respectively: (i) the effect of possible degradation products released from the materials to the culture medium and (ii) the effect of the surface properties on the osteoblast-like cells. In addition, the morphology of cells adherent to the surface of the starch-based polymers was analysed and correlated with their topography and with other chemical properties previously evaluated. The proliferation rate was found to differ from blend to blend as well as with the time of culture and with the presence of HA depending on the material. SEVA-C and respective composites systematically presented the higher number of cells comparatively to the other two blends. SPCL composites were found to be less suitable for cell proliferation. The amount of cells quantified after 7 days of culture, both on the surface and on the wells showed a delay in the proliferation of the cells cultured with SPCL composites comparatively to other materials and to TCPS. SCA composites, however, did support cell adhesion but also induce a slight level of toxicity, which results in delayed proliferation on the cells adhered to the wells. Cell morphology on the surface of the materials was also, in almost every case, found to be appropriate. In fact, cells were well adhered and spread on the majority of the surfaces. Thus, starch-based biomaterials can be seen as good substrates for osteoblast-adhesion and proliferation that demonstrates their potential to be used in orthopaedic applications and as bone tissue engineering scaffolds.Fundação para a Ciência e a Tecnologia (FCT

Physicochemical Properties of Gold Nanostructures Deposited on Glass

Properties of gold films sputtered onto borosilicate glass substrate were studied. UV-Vis absorption spectra were used to investigate optical parameters. XRD analysis provided information about the gold crystalline nanostructure, the texture, and lattice parameter and biaxial tension was also determined by the XRD method. The surface morphology was examined by atomic force microscopy (AFM); chemical structure of sputtered gold nanostructures was examined by X-ray photoelectron spectroscopy (ARXPS). The gold crystallites are preferentially [111] oriented on the sputtered samples. Gold deposition leads to dramatic changes in the surface morphology in comparison to pristine glass substrate. Oxygen is not incorporated into the gold layer during gold deposition. Experimental data on lattice parameter were also confirmed by theoretical investigations of nanoclusters using tight-binding potentials

Plasma activated PDMS microstructured pattern with collagen for improved myoblast cell guidance

We focused on polydimethylsiloxane (PDMS) as a substrate for replication, micropatterning, and construction of biologically active surfaces. The novelty of this study is based on the combina-tion of argon plasma exposure of micropatterned PDMS scaffold, where the plasma served as a strong tool for subsequent grafting of collagen coating and their application as cell growth scaf-folds, where the standard has been significantly exceeded. As part of scaffold design, templates with a patterned microstructure of different dimensions (50 x 50, 50 x 20 and 30 x 30 microns were created by photolithography followed by pattern replication on a PDMS polymer substrate. Subsequently, the prepared microstructured PDMS replicas were coated with a type I collagen layer. The sample preparation was followed by the characterization of material surface properties through various analytical techniques, including scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray photoelectron spectroscopy (XPS). To evaluate the biocompatibility of the produced samples, we conducted studies on the interactions between selected polymer replicas with micro- and nanostructures and mammalian cells. Specif-ically, we utilized mouse myoblasts (C2C12) and our results demonstrate that we achieved excellent cell alignment in conjunction with the development of a cytocompatible surface. Consequently, the outcomes of this research contribute to an enhanced comprehension of surface properties and interactions between structured polymers and mammalian cells. The use of periodic microstructures holds the potential for advancing the creation of novel materials and scaffolds in tissue engineering. These materials exhibit exceptional biocompatibility and possess the capacity to promote cell adhesion and growth

Surface activation of Hastalex by vacuum argon plasma for cytocompatibility enhancement

Here, we present surface analysis and biocompatibility evaluation of novel composite material based on graphene oxide traded as Hastalex. First, the surface morphology and elemental analysis of the pristine material were examined by atomic force and scanning electron microscopies, and by energy-dispersive and X-ray photoelectron spectroscopies, respectively. The Hastalex surface was then modified by plasma, 3 and 8 W with exposure times up to 240 s, the impact of which on the material surface wettability and morphology was further evaluated. In addition, the material aging was studied at room and elevated temperatures. Significant changes in surface roughness, morphology, and area were detected at the nanometre scale after plasma exposure. An increase in oxygen content due to the plasma exposure was observed both for 3 and 8 W. The plasma treatment had an outstanding effect on the cytocompatibility of Hastalex foil treated at both input powers of 3 and 8 W. The cell number of human MRC 5 fibroblasts on Hastalex foils exposed to plasma increased significantly compared to pristine Hastalex and even to tissue culture polystyrene. The plasma exposure also affected the fibroblasts cell growth and shape

LIPSS pattern induced by polymer surface instability for myoblast cell guidance

The presented study highlights the efficiency of employing a KrF excimer laser to create diverse types of periodic nanostructures (LIPSS - laser induced periodic surface structures) on polyether ether ketone (PEEK) and polyethylene naphthalate (PEN) substrates. By exposing the polymer films below their ablation threshold to laser fluence ranging from 4 to 16 mJcm-2 at 6,000 pulses, we studied both single-phase exposure at beam incidence angles of 0deg and 45deg, and two-phase exposure. Atomic force microscopy analysis revealed that the laser-treated samples contained distinctive periodic patterns such as waves, globules, and pod-like structures each exhibiting unique surface roughness. Moreover, using analytical methods like EDS and XPS shed light on the changes in the atomic composition, specifically focusing on the C and O elements, as a result of laser exposure. Notably, in almost all cases, we observed an increase in oxygen percentage on the sample surfaces. This increase not only led to a decrease in the contact angle with water but also lowered the zeta potential value, thus showing that the modified samples have enhanced hydrophilicity of the surface and altered electrostatic properties. Last but not least, the samples were assessed for biocompatibility; we studied the interaction of the prepared replicates with mouse myoblasts (C2C12). The impact of globular/dot structures on the cell growth in comparison to pristine or linear LIPSS-patterned surfaces was determined. The linear pattern (LIPSS) induced the myoblast cell alignment along the pattern direction, while dot/globular pattern even enhanced the cytocompatibility compared to LIPSS samples