Surface Modification of PLLA, PTFE and PVDF with Extreme Ultraviolet (EUV) to Enhance Cell Adhesion.

Recently, extreme ultraviolet (EUV) radiation has been increasingly used to modify polymers. Properties such as the extremely short absorption lengths in polymers and the very strong interaction of EUV photons with materials may play a key role in achieving new biomaterials. The purpose of the study was to examine the impact of EUV radiation on cell adhesion to the surface of modified polymers that are widely used in medicine: poly(tetrafluoroethylene) (PTFE), poly (vinylidene fluoride) (PVDF), and poly-L-(lactic acid) (PLLA). After EUV surface modification, which has been performed using a home-made laboratory system, changes in surface wettability, morphology, chemical composition and cell adhesion polymers were analyzed. For each of the three polymers, the EUV radiation differently effects the process of endothelial cell adhesion, dependent of the parameters applied in the modification process. In the case of PVDF and PTFE, higher cell number and cellular coverage were obtained after EUV radiation with oxygen. In the case of PLLA, better results were obtained for EUV modification with nitrogen. For all three polymers tested, significant improvements in endothelial cell adhesion after EUV modification have been demonstrated.

Calculation of water equivalent ratios for various materials at proton energies ranging 10-500 MeV using MCNP, FLUKA, and GEANT4 Monte Carlo codes.

Dosimetry of proton beams is generally evaluated in liquid water, or alternatively in solid phantoms via water equivalent ratios (WER). WER is defined as the ratio of proton range in liquid water to that in a phantom of certain material. Presently, WER is not available in the literature neither for a wide range of energies nor for variety of relevant materials. Thus, the goal of this study is to provide such data through Monte Carlo simulations. WER is calculated for 10-500 MeV energies for compact bone, adipose tissue, polymethyl methacrylate (PMMA), PTFE (teflon), graphite (C), aluminum (Al), copper (Cu), titanium (Ti), and gold (Au) using MCNPX.2.70, GEANT4, and FLUKA Monte Carlo (MC) codes. The MCNPX code was considered as the reference to which other codes were compared. The mean values of WER obtained through the MCNPX simulations for Au, Cu, Ti, Al, PTFE, graphite, PMMA, bone, and adipose tissue were 8.83, 5.40, 3.18, 2.03, 1.87, 1.52, 1.13, 1.71, and 0.96, respectively, for 10-500 MeV energy range. The maximum deviations of WER values between MCNPX and GEANT4 results were about 6.85% for adipose tissue at energies <20 MeV, whereas they were about 7.74%, 7.74% between MCNPX and FLUKA, for adipose and Al, respectively. This inter-code uncertainties are mainly due to different physic models and stopping powers in each code. Comparing the results to that in the literature, the range of discrepancy was found to be 0-8% with greatest discrepancy for Au. Based on the materials evaluated, the PMMA remained the closest to water, for a non-tissue solid material, with an average WER of 1.13, for proton energy ranging 10-500 MeV.

Mechanisms and Characterization of the Pulsed Electron-Induced Grafting of Styrene onto Poly(tetrafluoroethylene-co-hexafluoropropylene) to Prepare a Polymer Electrolyte Membrane.

During the pulsed-electron beam direct grafting of neat styrene onto poly(tetrafluoroethylene-co-hexafluoropropylene) (FEP) substrate, the radiolytically-produced styryl and carbon-centered FEP radicals undergo various desired and undesired competing reactions. In this study, a high-dose rate is used to impede the undesired free radical homopolymerization of styrene and ensure uniform covalent grafting through 125-µm FEP films. This outweighs the enhancement of the undesired crosslinking reactions of carbon-centered FEP radicals and the dimerization of the styryl radicals. The degree of uniform grafting through 125-µm FEP films increases from ≈8%, immediately after pulsed electron irradiation to 33% with the subsequent thermal treatment exceeding the glass transition temperature of FEP of 39°C. On the contrary, steady-state radiolysis using 60Co gamma radiolysis, shows that the undesired homopolymerization of the styrene has become the predominant reaction with a negligible degree of grafting. Time-resolved fast kinetic measurements on pulsed neat styrene show that the styryl radicals undergo fast decays via propagation homopolymerization and termination reactions at an observed reaction rate constant of 5 × 108 l · mol-1 · s-1. The proton conductivity of 25-µm film at 80°C is 0.29 ± 0.01 s cm-1 and 0.007 s cm-1 at relative humidity of 92% and 28%, respectively. The aims of this work are: 1. electrolyte membranes are prepared via grafting initiated by a pulsed electron beam; 2. postirradiation heat-treated membranes are uniformly grafted, ideal for industry; 3. High dose rate is the primary parameter to promote the desired reactions; 4. measurement of kinetics of undesired radiation-induced styrene homopolymerization; and 5. The conductivity of prepared membranes is on par or higher than industry standards.

Friction coefficient dependence on electrostatic tribocharging.

Friction between dielectric surfaces produces patterns of fixed, stable electric charges that in turn contribute electrostatic components to surface interactions between the contacting solids. The literature presents a wealth of information on the electronic contributions to friction in metals and semiconductors but the effect of triboelectricity on friction coefficients of dielectrics is as yet poorly defined and understood. In this work, friction coefficients were measured on tribocharged polytetrafluoroethylene (PTFE), using three different techniques. As a result, friction coefficients at the macro- and nanoscales increase many-fold when PTFE surfaces are tribocharged, but this effect is eliminated by silanization of glass spheres rolling on PTFE. In conclusion, tribocharging may supersede all other contributions to macro- and nanoscale friction coefficients in PTFE and probably in other insulating polymers.

Analysis of thickness of a hydrophobic fluoropolymer film based on electrowetting.

The electrowetting of water drops on a dielectric fluoropolymer film was studied experimentally. The dependence of the contact angles of the water drops on the applied voltage has been well explained in the low-voltage limit by using the classical Young-Lippmann theory. With this theory, the thicknesses of films coated on glass substrates by using a spin-coater were obtained indirectly by fitting the contact angle data and were confirmed by using X-ray reflectometry. The two sets of results showed a good agreement. In addition, we confirmed that the contact angle saturation at high voltage were consistent with Peykov's model.

External beam radiation therapy for PTFE dialysis grafts: a pilot study.

PURPOSE: The aim of this study was to identify the effects of external beam radiation on PTFE dialysis graft dysfunction. METHODS: Seven patients who underwent PTFE dialysis graft angioplasty were randomized to receive either two 8 Gy doses of external beam radiation or no radiation. The primary endpoint was time to graft thrombosis with a secondary endpoint of time to first intervention. RESULTS: There was no statistically significant difference between the two groups in either of the endpoints, although grafts in the radiation group had a shorter time to thrombosis or intervention. CONCLUSIONS: Our results demonstrate technical feasibility for use of external beam radiation in the setting of dialysis vascular access graft dysfunction. Larger randomized studies are required to identify whether there is a clinical benefit from this intervention.

UVC fluencies for preventative treatment of Pseudomonas aeruginosa contaminated polymer tubes.

Exposing Pseudomonas aeruginosa biofilm grown on the inner surface of Teflon and silicone tubes to UVC light (265 nm) from light emitting diodes (LED) has previously been shown to substantially reduce biofilm growth. Smaller UVC fluencies were required to disinfect Teflon tubes compared to silicone tubes. Light propagation enhancement in tubes can be obtained if the refractive index of the intra-luminal saline solution is higher than that of the polymer. This condition is achieved by using Teflon tubes with a low refractive index (1.34) instead of the polymers with a high refractive index (1.40-1.50) normally used for tubing in catheter production. Determining whether or not UVC light exposure can disinfect and maintain the intra-luminal number of colony forming units (CFUs) at an exceedingly low level and thus avoid the growth and establishment of biofilm is of interest. The use of UVC diodes is demonstrated to be a preventative disinfection treatment on tubes made of Teflon, which enhances the UVC light propagation, and on tubes made of a softer material, ethylene vinyl acetate (EVA), which is suitable for catheters but much less suitable for UVC light propagation. Simulating an aseptic breach (∼10(3)-10(4) CFU ml(-1)), the UVC disinfection set-up was demonstrated using tubes contaminated with planktonic P. aeruginosa. After the tubes (10-20 cm) were inoculated with the bacterial solution for 3 h, they were emptied and filled with saline solutions (0.9-20%). Next UVC fluencies (0-21 mJ cm(-2)) were applied to the tubes 3 h after inoculation. Colony counts were carried out on liquid samples drawn from the tubes the first day after UVC treatment and liquid and surface samples were collected and analyzed 3-4 days later. A fluence of approximately 1.0 mJ cm(-2) was noted as being sufficient for no growth for a period of 3-4 days for the Teflon tubes. Determining the fluence threshold for the EVA tubes was not possible. Almost all of the UVC-treated EVA tubes were disinfected simply by filling the tubes with a saline solution. Direct UVC treatment of the contaminated EVA tubes revealed, however, that a fluence of 21 mJ cm(-2) killed the bacteria present in the tubes and kept them disinfected for a period of 3-4 days.

Effectiveness of different substrate materials for in vitro sunscreen tests.

BACKGROUND: In vitro measurements of sunscreen products are used to assess their reliability in terms of photoprotection and photo-stability. OBJECTIVE: In this study, several substrates have been fully characterized for in vitro sunscreen testing. METHODS: The following different substrates have been utilised in the study: roughened PMMA plates, Transpore, Vitro-Skin, roughened quartz plate and a laboratory prepared roughened PTFE plate. The suitability of these substrates for SPF evaluation has been investigated by performing total absorbance measurements of seven sunscreen products with different SPF values produced by the same manufacturer. RESULTS AND CONCLUSIONS: Application of sunscreen products on Transpore, roughened quartz plate, PMMA, PTFE and Vitro-Skin was performed efficiently. With regard to photo-stability of the substrate materials, only PMMA plate, PTFE and roughened quartz plate showed to be photo-stable after UV irradiation. With regard to SPF tests, our results indicate that Transpore, roughened quartz plate and Vitro-Skin are preferable to assess SPF values because of a better correlation between in vitro and in vivo measurements. Our study also confirms that an initial calibration must be performed for sunscreen products labelled with different SPF values. Finally, the results of our measurements demonstrate that, although a correlation between in vitro and in vivo SPF results can be established, it is never exactly 1:1.

Beam-surface scattering studies of the individual and combined effects of VUV radiation and hyperthermal O, O2, or Ar on FEP Teflon surfaces.

Beam-surface scattering experiments were used to probe products that scattered from FEP Teflon surfaces during bombardment by various combinations of atomic and molecular oxygen, Ar atoms, and vacuum ultraviolet (VUV) light. A laser-breakdown source was used to create hyperthermal (translational energies in the range 4-13 eV) beams of argon and atomic/molecular oxygen. The average incidence energy of these beams was tunable and was controlled precisely with a synchronized chopper wheel. A filtered deuterium lamp provided a source of VUV light in a narrow-wavelength range centered at 161 nm. Volatile products that exited the surfaces were monitored with a rotatable mass spectrometer detector. Hyperthermal O atoms with average translational energies above approximately 4 eV may react directly with a pristine FEP Teflon surface, and the reactivity appears to increase with the translational energy of the incident O atoms. VUV light or highly energetic collisions of O2 or Ar may break chemical bonds and lead to the ejection of volatile products; the ejection of volatile products is enhanced when the surface is subjected to VUV light and energetic collisions simultaneously. Exposure to VUV light or to hyperthermal O2 or Ar may increase the reactivity of an FEP Teflon surface to O atoms.

Technical report: effects of PUVA treatment on the optical properties of blood/tissue storage bags during extracorporeal photochemotherapy.

Extracorporeal photochemotherapy (photopheresis, ECP) is a novel therapeutic method for patients who do not respond to immunosuppressive medications, and gaining interest in the treatment of Graft-vs-Host Disease. This paper is focused on the optical transmission properties of plastic bags which can be used in an independent (off-line) method of ECP, and reports the results of spectral measurements on various bags of different chemical compositions, with and without PUVA treatment. Regarding their higher and more uniform UVA transmission values, FEP based bags perform superior to the others. Considering its UVB absorption and UVA transmission properties, the EVA bag is a good choice, while Polyimide Kapton-FEP plastic film should not be considered for use in ECP. PUVA treatment of blood bags may affect their optical behaviour, and causes reduction of transmission of the material in UV range of the spectrum.

Analysis of photoinitiated polymerization in a membrane mimetic film using infrared spectroscopy and near-IR Raman microscopy.

A method has been developed to investigate the extent of polymer cross-linking that results following in situ photopolymerization of an acrylate-functionalized phospholipid assembly adsorbed onto a stabilized, membrane-mimetic film produced from a polyelectrolyte multilayer (PEM) on polytetrafluoroethylene (PTFE) grafts. The acrylate phospholipid monomer was synthesized, prepared as a unilamellar vesicle, and fused onto closed-packed acyl chains that make up the PEM membrane-mimetic barrier on the PTFE graft. Both broad band white light and 514.5 nm laser radiation were used as excitation sources for photoinitiation; eosin Y was used as the photoinitiator. The use of 514.5 nm excitation reduced the time for maximum polymerization of the acrylate lipid from 60 min to 240 s. Infrared spectroscopy was successfully used to analyze the extent of photopolymerization in simplified model acrylate lipid systems; however, this method could not be used to analyze acrylate polymerization in heterogeneous, multicomponent PEM membrane-mimetic barriers on PTFE grafts. A near-infrared Raman microscopy method based on the ratio of the integrated areas of the CC and CN vibrations was shown to provide equivalent information to the IR method for analysis of the extent of polymerization efficiency in acrylate lipids. In addition, it proved feasible to extend this near-IR Raman method to the in situ analysis of the extent of polymerization in a stabilized acrylate lipid membrane on a PEM film in a PTFE vascular graft. This work describes a new approach for generating and analyzing the robustness of a membrane-mimetic coating on biomaterial surfaces, and may improve our ability to predict the long-term stability of polymeric membrane-mimetic films on implantable medical devices.

Cell microarrays on photochemically modified polytetrafluoroethylene.

We studied the adhesion, proliferation, and viability of human umbilical vein endothelial cells (HUVEC) and human embryonic kidney cells (HEK) on modified spots at polytetrafluoroethylene (PTFE) surfaces. The viability of the cells was assessed using an aqueous non-radioactive cell proliferation assay. Round spots with a diameter of 100 microm were modified by exposure to the ultraviolet (UV) light of a Xe(2)(*)-excimer lamp at a wavelength of 172 nm in an ammonia atmosphere employing a contact mask. The spots were arranged in a quadratic pattern with 300 microm center-to-center spot distances. With optimized degree of modification, the cells adhered to the modified spots with a high degree of selectivity (70-90%). The adhered cells on the spots proliferated. This resulted in a significant increase in the number of adhering HUVECS or HEK cells after seeding and in the formation of confluent cell clusters after 3-4 days. With higher start seeding density, these clusters were not only confined to the modified spots but extended several micrometer to the neighborhood. The high potential of the cell microarrays for gene analysis in living cells was demonstrated with HEK cells transfected by yellow fluorescent protein (YFP).

Immobilization of streptavidin-horseradish peroxidase onto a biotinylated poly(acrylic acid) backbone that had been radiation-grafted to a PTFE film.

Films of polytetrafluoroethylene (PTFE) were modified by radiation graft polymerization of acrylic acid (AAc). Optimal conditions for efficient AAc grafting were studied, including pre-irradiation dose in air, monomer concentration, temperature and time of the grafting process. Carboxylic groups of the grafted polyAAc were activated with carbodiimide (EDC) for biotinylation by reaction with 5-(biotinamido) pentylamine. Streptavidin-horseradish peroxidase (SA-HRP) was immobilized by affinity complexation of the SA with the biotin groups on the PTFE surface. The amount of active HRP immobilized on the PTFE films was determined as a function of the extent of polyAAc grafting. This study has demonstrated the utility of combining the processes of (a) radiation grafting of polymers with reactive groups onto inert polymers such as PTFE, (b) biotinylation of the graft polymer reactive groups, (c) immobilization of streptavidin on the biotinylated surface sites, followed by (d) immobilization of biotinylated, biologically active molecules via complexation of their conjugates with streptavidin. In this study, the last two steps were combined by immobilizing the complex of streptavidin and biotinylated HRP onto the biotinylated surface sites. The unique nature of this process is the ability to immobilize biotinylated molecules on an inert surface as PTFE.

[Ion-beam irradiated ePTFE for the therapy of intracranial aneurysms].

Intracranial aneurysms are frequently treated with either microsurgical clipping or endovascular coiling. However, as so called broad-neck aneurysms are not suitable for these treatment options, a wrapping technique using muslin gauze, muscle piece, ePTFE is applied for such cases. The material for aneurysmal wrapping demands both stable adherence and no reactive inflammatory response such as inert artificial wall. Authors have developed a new improved ePTFE by ion-beam irradiation technique that is biologically inert and able to adhere firmly to surrounding tissue. Based on the last studies, Ar+ ion at an energy of 150 keV with a fluence of 5 x 10(14) ions/cm2 was chosen to irradiate ePTFE. A cell adhesion test and direct implantation of ion-beam irradiated ePTFE as wrapping material to rabbit common carotid arteries (CCA) were examined. It was demonstrated that the surface of ion-beam irradiated ePTFE exhibits remarkably greater adhesion and promotes cell proliferation on the surface more effectively than that of non-irradiated ePTFE. The carotid artery well-wrapped by ion-beam irradiated ePTFE strongly adhered to the mural wall and induced little inflammatory reaction. The results of this investigation indicate that application of this technology would offer the best means for aneurysm wrapping.

[Biocompatibility of ePTFE modified by ion beam irradiation].

Expanded polytetrafluoroethylene (ePTFE) is a stable polymer and widely used as a prosthesis because of its chemical inertness. However, ePTFE as an artificial dura mater is often associated with postoperative leakage of the cerebrospinal fluid (CSF) due to its very low adhesiveness to fibrin glue and surrounding tissue. To overcome this shortcoming the authors examined the effects of ion beam irradiation to ePTFE surface. The surfaces of ePTFE were irradiated with 150keV--He+, Ne-, Ar+ and Kr+ ions with fluences of 1 x 10(14), 5 x 10(14) and 1 x 10(15) ions/cm2. L929 fibroblasts were cultured for 24 hours on ePTFE sheets that had both ion beam irradiated and un-irradiated regions. It was confirmed that fibroblasts still adhered to the ion beam irradiated area. This phenomenon was observed under different condition of ion beam irradiation. Adhesive strength of fibrin glue to ion beam irradiated ePTFE was evaluated by tensile strength and burst pressure test using ePTFE patch. Both tensile strength and burst pressure were remarkably enhanced by ion beam irradiation. SEM study indicated that fibrin glue infiltrated and anchored into the gaps induced by the ion beam irradiation. Thus ion beam irradiation improved biocompatibility of the surface of ePTFE.

[Ion-beam irradiated ePTFE for an artificial dura mater].

Expanded polytetrafluoroethylene (ePTFE) is a stable polymer and widely used as a prosthesis because of its chemical inertness. However, ePTFE as an artificial dura mater is often associated with postoperative leakage of the cerebrospinal fluid (CSF) due to its very low adhesiveness to fibrin glue and surrounding tissue. The surfaces of ePTFE were modified by ion-beam irradiation in order to improve tissue adhesiveness. The morphological changes of ePTFE that was irradiated with 150 keV-He+, Net, Ar+ and Kr+ ions applied at a rate of 1 x 10(14), 5 x 10(14) and 1 x 10(15) ions/cm2 were examined with scanning electron microscopy. Next, surrounding tissue reactions to the implanted ePTFE into the rabbit calvarial defects were analyzed at intervals from 1 week up to 1 month. The micromesh structures of the ePTFE were destroyed by ion-beam irradiation, and the depth and width of the gaps increased with heavy particle ion and irradiated fluence. In an implantation study using experimental rabbits, a dura defect was patched with four types of ion-beam irradiated ePTFE with different in irradiation fluence. A CSF leakage was observed in the rabbit covered with non-irradiated ePTFE, but, CSF leakage did not occur in any of the rabbits covered with ion-beam irradiated ePTFE that adhered in a water-tight manner to surrounding tissues. Histological study indicated that fibroblast-like cells had infiltrated and anchored the gaps in the ion-beam irradiated ePTFE. Ar+ or Kr+ with 1 x 10(15) ions/cm2 irradiated ePTFE had the highest adhesiveness and produced the best healing of dura defects.

Cell adhesion on polytetrafluoroethylene modified by UV-irradiation in an ammonia atmosphere.

We report on the modification of polytetrafluoroethylene (PTFE) by exposure to the ultraviolet (UV) light of a Xe(2)*-excimer lamp at a wavelength of 172 nm in an ammonia atmosphere. Typical treatment times were up to 30 min. Subsequently, the samples were grafted with the amino acid alanine from an aqueous solution. The samples were characterized by means of optical transmission spectroscopy, laser-induced fluorescence and contact-angle measurements. We studied the adhesion of rat aortic smooth muscle cells (SMC) and mouse fibroblasts (3T3 cells) to the modified polymer samples using an in vitro technique, where the population density and spread of adhering cells is determined 24 h after seeding by image analysis. For both cell types the exposure of PTFE to UV-light in an ammonia atmosphere resulted in a significant increase in the number of adhering cells and in the size of their spreading area. The grafting with alanine enhanced this effect. Additional experiments with human endothelial cells (HEC) also demonstrated improved adhesion to modified PTFE. Thus, PTFE modified by our method appears to be a promising material for fabrication of artificial vascular prostheses and implants or for cultivation of skin substitutes.

Adhesion and proliferation of human endothelial cells on photochemically modified polytetrafluoroethylene.

We studied the adhesion and proliferation of human endothelial cells on photochemically modified polytetrafluoroethylene samples. The polymer surfaces were modified by exposure to the ultraviolet light of a Xe(2)(*)-excimer lamp at a wavelength of 172 nm in an ammonia atmosphere. Treatment times were between 10 and 20 min. The endothelial cell density was determined 1, 3 and 8 days after seeding by image analysis. Surface modification of the samples resulted in a significant increase in the number of adhering cells and in the formation of a confluent cell layer after 3-8 days. The results were comparable than those obtained on polystyrene Petri dishes, which are used as standard substrates in cell cultivation. Thus modified PTFE appears to be a promising material for the fabrication of artificial vascular prostheses coated with endothelial cells.

Novel material properties via ionising radiation.

Ionising radiation is normally associated with the degradation of materials during radiation sterilisation. However, these same radiation sources can be used to create material properties that are unobtainable by other methods. This review of some novel properties that can be achieved also critically compares the relative merits and disadvantages of each radiation source.

High-speed MAS-NMR investigations on radiation-modified fluoropolymers.

Fluoropolymers possess an interesting combination of physical and chemical properties. In order to utilize these properties in combination with other materials, the compatibility of the fluoropolymers with those materials has to be improved by a suitable modification process. As one modification process the irradiation with high-energy electrons has been applied. High-speed MAS solid-state NMR has been used to characterize the structural changes in fluoropolymers resulting from the irradiation. The high MAS rotation frequencies, applied here, average the anisotropy of the chemical shift and the homonuclear dipolar coupling. Despite the fact that the electron irradiation results in a low concentration of modifications, the dynamic range of the solid-state NMR experiment is sufficient to detect and characterize the modifications.