Ultraviolet photofunctionalization of nanostructured titanium surfaces enhances thrombogenicity and platelet response.

The purpose of this study was to evaluate blood and platelet response to nanostructured TiO2 coatings and to investigate the effect of Ultraviolet (UV) light treatment on blood clotting ability, platelet activation and protein adhesion. Ti-6Al-4V titanium alloy plates (n = 138) were divided into three groups; a sol-gel derived MetAliveTM coating (MA); hydrothermal coating (HT); and a non-coated group (NC). Sixty nine titanium substrates were further treated with UV light for 1 h. The thrombogenicity of the titanium substrates was assessed using fresh human blood with a whole blood kinetic clotting time method. The platelet adhesion test was conducted to evaluate the morphology and adhesion behavior of the platelets on the titanium substrates. Human diluted plasma and bovine fibronectin were used to evaluate protein adsorption. Total clotting time for the UV treated HT, MA and NC titanium substrates was almost 40 min compared to 60 min for non-UV substrates, the total clotting time for the UV treated groups were significantly lower than that of the non UV NC group (p < 0.05). UV light treatment had significantly enhanced coagulation rates. The HT and MA substrates presented more platelet aggregation, spreading and pseudopod formation in comparison with the NC substrates. UV treatment did not affect the platelet activation and protein adsorption. This in vitro study concluded that nanostructured titanium dioxide implant surfaces obtained by sol-gel and hydrothermal coating methods increased coagulation rates and enhanced platelet response when compared with non-coated surfaces. UV light treatment clearly improved thrombogenicity of all examined Ti-6Al-4V surfaces.

UV-Curable Aliphatic Silicone Acrylate Organic-Inorganic Hybrid Coatings with Antibacterial Activity.

The most effective means to protect against bacterial invasion and to reduce the risk of healthcare-associated infections are antibacterial components synthesis. In this study, a novel process for the synthesis of organic-inorganic hybrid coatings containing silver nanoparticles is presented. Silver nanoparticles and polymer formation proceeds simultaneously through the in situ photoreduction of silver salt to silver nanoparticles and UV-crosslinking of bifunctional aliphatic silicone acrylate. The nanocomposite films with 0.5-1.43 wt % of silver nanoparticles concentration were obtained and investigated. The formation of silver nanoparticles in polymer matrix was confirmed via UV-visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy, scanning electron spectroscopy, and energy dispersive spectroscopy. Our investigations clearly show the formation of silver nanoparticles in silicone acrylate network. Direct photoreduction of silver salt by UV-radiation in the organic media produced silver nanoparticles exhibiting cubic crystal structure. The size of nanoparticles was determined to be near 20 ± 5 nm. The antibacterial activities of coatings were determined using the disc diffusion and direct contact methods. UV-curable silicone acrylate hybrid coatings exhibited antibacterial activity against harmful bacteria strains.

Effective protection of biological membranes against photo-oxidative damage: Polymeric antioxidant forming a protecting shield over the membrane.

We have prepared a chitosan polymer modified with gallic acid in order to develop an efficient protection strategy biological membranes against photodamage. Lipid bilayers were challenged with photoinduced damage by photosensitization with methylene blue, which usually causes formation of hydroperoxides, increasing area per lipid, and afterwards allowing leakage of internal materials. The damage was delayed by a solution of gallic acid in a concentration dependent manner, but further suppressed by the polymer at very low concentrations. The membrane of giant unilamellar vesicles was covered with this modified macromolecule leading to a powerful shield against singlet oxygen and thus effectively protecting the lipid membrane from oxidative stress. The results have proven the discovery of a promising strategy for photo protection of biological membranes.

Design of Albumin-Coated Microbubbles Loaded With Polylactide Nanoparticles.

OBJECTIVES: A protocol was designed to produce albumin-coated microbubbles (MBs) loaded with functionalized polylactide (PLA) nanoparticles (NPs) for future drug delivery studies. METHODS: Microbubbles resulted from the sonication of 5% bovine serum albumin and 15% dextrose solution. Functionalized NPs were produced by mixing fluorescent PLA and PLA-polyethylene glycol-carboxylate conjugates. Nanoparticle-loaded MBs resulted from the covalent conjugation of functionalized NPs and MBs. Three NP/MB volume ratios (1/1, 1/10, and 1/100) and unloaded MBs were produced and compared. Statistical evaluations were based on quantitative analysis of 3 parameters at 4 time points (1, 4, 5, and 6 days post MB fabrication): MB diameter using a circle detection routine based on the Hough transform, MB number density using a hemocytometer, and NP-loading yield based on MB counts from fluorescence and light microscopic images. Loading capacity of the albumin-coated MBs was evaluated by fluorescence. RESULTS: Loaded MB sizes were stable over 6 days after production and were not significantly different from that of time-matched unloaded MBs. Number density evaluation showed that only 1/1 NP/MB volume ratio and unloaded MB number densities were stable over time, and that the 1/1 MB number density evaluated at each time point was not significantly different from that of unloaded MBs. The 1/10 and 1/100 NP/MB volume ratios had unstable number densities that were significantly different from that of unloaded MBs (P < .05). Fluorescence evaluation suggested that 1/1 MBs had a higher NP-loading yield than 1/10 and 1/100 MBs. Quantitative loading evaluation suggested that the 1/1 MBs had a loading capacity of 3700 NPs/MB. CONCLUSIONS: A protocol was developed to load albumin MBs with functionalized PLA NPs for further drug delivery studies. The 1/1 NP/MB volume ratio appeared to be the most efficient to produce stable loaded MBs with a loading capacity of 3700 NPs/MB.

Preventing bacterial growth on implanted device with an interfacial metallic film and penetrating X-rays.

Device-related infections have been a big problem for a long time. This paper describes a new method to inhibit bacterial growth on implanted device with tissue-penetrating X-ray radiation, where a thin metallic film deposited on the device is used as a radio-sensitizing film for bacterial inhibition. At a given dose of X-ray, the bacterial viability decreases as the thickness of metal film (bismuth) increases. The bacterial viability decreases with X-ray dose increases. At X-ray dose of 2.5 Gy, 98% of bacteria on 10 nm thick bismuth film are killed; while it is only 25% of bacteria are killed on the bare petri dish. The same dose of X-ray kills 8% fibroblast cells that are within a short distance from bismuth film (4 mm). These results suggest that penetrating X-rays can kill bacteria on bismuth thin film deposited on surface of implant device efficiently.

Biofunctionalization strategies on tantalum-based materials for osseointegrative applications.

The use of tantalum as biomaterial for orthopedic applications is gaining considerable attention in the clinical practice because it presents an excellent chemical stability, body fluid resistance, biocompatibility, and it is more osteoconductive than titanium or cobalt-chromium alloys. Nonetheless, metallic biomaterials are commonly bioinert and may not provide fast and long-lasting interactions with surrounding tissues. The use of short cell adhesive peptides derived from the extracellular matrix has shown to improve cell adhesion and accelerate the implant's biointegration in vivo. However, this strategy has been rarely applied to tantalum materials. In this work, we have studied two immobilization strategies (physical adsorption and covalent binding via silanization) to functionalize tantalum surfaces with a cell adhesive RGD peptide. Surfaces were used untreated or activated with either HNO3 or UV/ozone treatments. The process of biofunctionalization was characterized by means of physicochemical and biological methods. Physisorption of the RGD peptide on control and HNO3-treated tantalum surfaces significantly enhanced the attachment and spreading of osteoblast-like cells; however, no effect on cell adhesion was observed in ozone-treated samples. This effect was attributed to the inefficient binding of the peptide on these highly hydrophilic surfaces, as evidenced by contact angle measurements and X-ray photoelectron spectroscopy. In contrast, activation of tantalum with UV/ozone proved to be the most efficient method to support silanization and subsequent peptide attachment, displaying the highest values of cell adhesion. This study demonstrates that both physical adsorption and silanization are feasible methods to immobilize peptides onto tantalum-based materials, providing them with superior bioactivity.

A simple one pot purification of bacterial amylase from fermented broth based on affinity toward starch-functionalized magnetic nanoparticle.

Surface-functionalized adsorbant particles in combination with magnetic separation techniques have received considerable attention in recent years. Selective manipulation on such magnetic nanoparticles permits separation with high affinity in the presence of other suspended solids. Amylase is used extensively in food and allied industries. Purification of amylase from bacterial sources is a matter of concern because most of the industrial need for amylase is met by microbial sources. Here we report a simple, cost-effective, one-pot purification technique for bacterial amylase directly from fermented broth of Bacillus megaterium utilizing starch-coated superparamagnetic iron oxide nanoparticles (SPION). SPION was prepared by co-precipitation method and then functionalized by starch coating. The synthesized nanoparticles were characterized by transmission electron microscopy (TEM), a superconducting quantum interference device (SQUID, zeta potential, and ultraviolet-visible (UV-vis) and Fourier-transform infrared (FTIR) spectroscopy. The starch-coated nanoparticles efficiently purified amylase from bacterial fermented broth with 93.22% recovery and 12.57-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the molecular mass of the purified amylase was 67 kD, and native gel showed the retention of amylase activity even after purification. Optimum pH and temperature of the purified amylase were 7 and 50°C, respectively, and it was stable over a range of 20°C to 50°C. Hence, an improved one-pot bacterial amylase purification method was developed using starch-coated SPION.

Direct patterning of poly(acrylic acid) on polymer surfaces by ion beam lithography for the controlled adhesion of mammalian cells.

Poly(acrylic acid) (PAA)-patterned polystyrene (PS) substrates were prepared by ion beam lithography to control cell behaviors of mouse fibroblasts and human embryonic kidney cells. Thin PAA films spin-coated on non-biological PS substrates were selectively irradiated with energetic proton ions through a pattern mask. The irradiated substrates were developed with deionized water to generate negative-type PAA patterns. The surface characteristics of the resulting PAA-patterned PS surface, such as surface morphology, chemical structure and composition and wettability, were investigated. Well-defined 100 µm PAA patterns were effectively formed on relatively hydrophobic PS substrates by ion beam lithography at higher fluences than 5 × 10(14) ions/cm(2). Moreover, based on the in vitro cell culture test, cells were adhered and proliferated favorably onto hydrophilic PAA regions separated by hydrophobic PS regions on the PAA-patterned PS substrates, and thereby leading to the formation of well-defined cell patterns.

Tuning DNA binding kinetics in an optical trap by plasmonic nanoparticle heating.

We report on the tuning of specific binding of DNA attached to gold nanoparticles at the individual particle pair (dimer) level in an optical trap by means of plasmonic heating. DNA hybridization events are detected optically by the change in the plasmon resonance frequency due to plasmonic coupling of the nanoparticles. We find that at larger trapping powers (i.e., larger temperatures and stiffer traps) the hybridization rates decrease by more than an order of magnitude. This result is explained by higher temperatures preventing the formation of dimers with lower binding energies. Our results demonstrate that plasmonic heating can be used to fine tune the kinetics of biomolecular binding events.

Evaluation of experimental coating to improve the zirconia-veneering ceramic bond strength.

PURPOSE: To evaluate the shear bond strength (SBS) between zirconia and veneering ceramic following different surface treatments of zirconia. The efficacy of an experimental zirconia coating to improve the bond strength was also evaluated. MATERIALS AND METHODS: Zirconia strips were fabricated and were divided into four groups as per their surface treatment: polished (control), airborne-particle abrasion, laser irradiation, and application of the experimental coating. The surface roughness and the residual monoclinic content were evaluated before and after the respective surface treatments. A scanning electron microscope (SEM) analysis of the experimental surfaces was performed. All specimens were subjected to shear force in a universal testing machine. The SBS values were analyzed with one-way ANOVA followed by Bonferroni post hoc for groupwise comparisons. The fractured specimens were examined to observe the failure mode. RESULTS: The SBS (29.17 MPa) and roughness values (0.80) of the experimental coating group were the highest among the groups. The residual monoclinic content was minimal (0.32) when compared to the remaining test groups. SEM analysis revealed a homogenous surface well adhered to an undamaged zirconia base. The other test groups showed destruction of the zirconia surface. The analysis of failure following bond strength testing showed entirely cohesive failures in the veneering ceramic in all study groups. CONCLUSION: The experimental zirconia surface coating is a simple technique to increase the microroughness of the zirconia surface, and thereby improve the SBS to the veneering ceramic. It results in the least monoclinic content and produces no structural damage to the zirconia substructure.

The effect of Er:YAG laser irradiation on hydroxyapatite-coated implants and fluoride-modified TiO2-blasted implant surfaces: a microstructural analysis.

The purpose of this study was to evaluate the microscopic changes and surface roughness on hydroxyapatite (HA)-coated implants following exposure to different powers and durations of Er:YAG laser irradiation in order to determine the proper pulse energy level and irradiation time. Ten HA-coated implants and ten fluoride-modified TiO2 implants were used. The implants were divided into a control (one implant) and test group (nine implants) for each implant type. Implants in the test groups were sub-divided into three groups (three implants per group) based on the applied laser pulse energy and irradiation time. The measurement of surface roughness was performed on all implants in the test groups using a white light interferometer before and after laser irradiation. R a values were recorded and compared in order to evaluate changes in surface roughness. For HA-coated implants, the R a values increased in all test groups after laser irradiation. However, mean R a values in the fluoride-modified TiO2-blasted implant test group were decreased after irradiation. There was no statistical difference. Scanning electron microscope analysis revealed surface alterations in both the HA-coated and fluoridated TiO2-blasted implants irradiated for 1.5 min at 100 mJ/pulse, 10 Hz. When the pulse energy and irradiation time increased, greater surface alterations, including surface flattening and microfractures, were observed. In conclusion, the results of the current study suggest that no changes could be observed in both HA-coated implants and fluoride-modified TiO2-blasted implants after irradiation at an intensity of 100 mJ/pulse, 10 Hz for 1 min performed to achieve surface detoxification.

LiNbO3 coating on concrete surface: a new and environmentally friendly route for artificial photosynthesis.

The addition of a photocatalyst to ordinary building materials such as concrete creates environmentally friendly materials by which air pollution or pollution of the surface can be diminished. The use of LiNbO3 photocatalyst in concrete material would be more beneficial since it can produce artificial photosynthesis in concrete. In these research photoassisted solid-gas phases reduction of carbon dioxide (artificial photosynthesis) was performed using a photocatalyst, LiNbO3, coated on concrete surface under illumination of UV-visible or sunlight and showed that LiNbO3 achieved high conversion of CO2 into products despite the low levels of band-gap light available. The high reaction efficiency of LiNbO3 is explained by its strong remnant polarization (70 µC/cm(2)), allowing a longer lifetime of photoinduced carriers as well as an alternative reaction pathway. Due to the ease of usage and good photocatalytic efficiency, the research work done showed its potential application in pollution prevention.

Carbon-centered radicals in γ-irradiated bone substituting biomaterials based on hydroxyapatite.

Gamma irradiated synthetic hydroxyapatite, bone substituting materials NanoBone(®) and HA Biocer were examined using EPR spectroscopy and compared with powdered human compact bone. In every case, radiation-induced carbon centered radicals were recorded, but their molecular structures and concentrations differed. In compact bone and synthetic hydroxyapatite the main signal assigned to the CO(2) (-) anion radical was stable, whereas the signal due to the CO(3) (3-) radical dominated in NanoBone(®) and HA Biocer just after irradiation. However, after a few days of storage of these samples, also a CO(2) (-) signal was recorded. The EPR study of irradiated compact bone and the synthetic graft materials suggest that their microscopic structures are different. In FT-IR spectra of NanoBone(®), HA Biocer and synthetic hydroxyapatite the HPO(4) (2-) and CO(3) (2-) in B-site groups are detected, whereas in compact bone signals due to collagen dominate.

NIR-Reflective and Hydrophobic Bio-Inspired Nano-Holed Configurations on Titanium Alloy.

Near-infrared (NIR) radiation plays an important role in guided external stimulus therapies; its application in bone-related treatments is becoming more and more frequent. Therefore, metallic biomaterials that exhibit properties activated by NIR are promising for further orthopedic procedures. In this work, we present an adapted electroforming approach to attain a biomorphic nano-holed TiO2 coating on Ti6Al4V alloy. Through a precise control of the anodization conditions, structures revealed the formation of localized nano-pores arranged in a periodic assembly. This specific organization provoked higher stability against thermal oxidation and precise hydrophobic wettability behavior according to Cassie-Baxter's model; both characteristics are a prerequisite to ensure a favorable biological response in an implantable structure for guided bone regeneration. In addition, the periodically arranged sub-wavelength-sized unit cell on the metallic-dielectric structure exhibits a peculiar optical response, which results in higher NIR reflectivity. Accordingly, we have proved that this effect enhances the efficiency of the scattering processes and provokes a significant improvement of light confinement producing a spontaneous NIR fluorescence emission. The combination of the already favorable mechanical and biocompatibility properties of Ti6Al4V, along with suitable thermal stability, wetting, and electro-optical behavior, opens a promising path toward strategic bone therapeutic procedures.

Laser surface modification for synthesis of textured bioactive and biocompatible Ca-P coatings on Ti-6Al-4V.

A textured calcium phosphate based bio-ceramic coating was synthesized by continuous wave Nd:YAG laser induced direct melting of hydroxyapatite precursor on Ti-6Al-4V substrate. Two different micro-textured patterns (100 µm and 200 µm line spacing) of Ca-P based phases were fabricated by this technique to understand the alignment and focal adhesion of the bone forming cells on these surfaces. X-ray diffraction studies of the coated samples indicated the presence of CaTiO3, α-Ca3(PO4)2, Ca(OH)2, TiO2 (anatase) and TiO2 (rutile) phases as a result of the intermixing between the precursor and substrate material during laser processing. A two dimensional elemental mapping of the cross-section of the coated samples exhibited the presence of higher phosphorous concentration within the coating and a thin layer of calcium concentration only at the top of the coating. Improved in vitro bioactivity and in vitro biocompatibility was observed for the laser processed samples as compared to the control.

Universal Antifouling and Photothermal Antibacterial Surfaces Based on Multifunctional Metal-Phenolic Networks for Prevention of Biofilm Formation.

Biofilms formed from the pathogenic bacteria that attach to the surfaces of biomedical devices and implantable materials result in various persistent and chronic bacterial infections, posing serious threats to human health. Compared to the elimination of matured biofilms, prevention of the formation of biofilms is expected to be a more effective way for the treatment of biofilm-associated infections. Herein, we develop a facile method for endowing diverse substrates with long-term antibiofilm property by deposition of a hybrid film composed of tannic acid/Cu ion (TA/Cu) complex and poly(ethylene glycol) (PEG). In this system, the TA/Cu complex acts as a multifunctional building block with three different roles: (i) as a versatile "glue" with universal adherent property for substrate modification, (ii) as a photothermal biocidal agent for bacterial elimination under irradiation of near-infrared (NIR) laser, and (iii) as a potent linker for immobilization of PEG with inherent antifouling property to inhibit adhesion and accumulation of bacteria. The resulted hybrid film shows negligible cytotoxicity and good histocompatibility and could prevent biofilm formation for at least 15 days in vitro and suppress bacterial infection in vivo, showing great potential for practical applications to solve the biofilm-associated problems of biomedical materials and devices.

Frictional properties of light-activated antimicrobial polymers in blood vessels.

The adhesion of microbes to catheter surfaces is a serious problem and the resulting infections frequently lead to longer hospitalisation and higher risk for the patient. Several approaches have been developed to produce materials that are less susceptible to microbial colonisation. One such approach is the incorporation of photoactivated compounds, such as Toluidine Blue O (TBO), in the polymeric matrix resulting in 'light-activated antimicrobial materials'. The insertion and removal of catheters can cause tissue damage and patient discomfort through frictional forces; hence the lubricity of a catheter material is also very important. In this work the tribological performance of silicone and polyurethane containing TBO and gold nanoparticles were evaluated using two different surfaces, the inner part of the aorta and the superior vena cava of sheep. Static and kinetic friction coefficients of these materials were measured using a tribometric device developed for in vitro applications using dry materials and those lubricated with blood. It was found that neither the preparation process nor the presence of TBO or gold nanoparticles, had an effect on the friction factors in comparison to those of untreated materials. In all cases, static and kinetic friction coefficients on aorta tissue were higher than those on vena cava due to higher surface roughness of the aorta. The presence of blood as a lubricant resulted in lower friction coefficients.

Ultrasound-assisted powder-coating technique to improve content uniformity of low-dose solid dosage forms.

An ultrasound-assisted powder-coating technique was used to produce a homogeneous powder formulation of a low-dose active pharmaceutical ingredient (API). The powdered particles of microcrystalline cellulose (MCC; Avicel® PH-200) were coated with a 4% m/V aqueous solution of riboflavin sodium phosphate, producing a uniform drug layer on the particle surfaces. It was possible to regulate the amount of API in the treated powder. The thickness of the API layer on the surface of the MCC particles increased near linearly as the number of coating cycles increased, allowing a precise control of the drug content. The tablets (n = 950) prepared from the coated powder showed significantly improved weight and content uniformity in comparison with the reference tablets compressed from a physical binary powder mixture. This was due to the coated formulation remaining uniform during the entire tabletting process, whereas the physical mixture of the powders was subject to segregation. In conclusion, the ultrasound-assisted technique presented here is an effective tool for homogeneous drug coating of powders of irregular particle shape and broad particle size distribution, improving content uniformity of low-dose API in tablets, and consequently, ensuring the safe delivery of a potent active substance to patients.

Nano-coating of beta-galactosidase onto the surface of lactose by using an ultrasound-assisted technique.

We nano-coated powdered lactose particles with the enzyme beta-galactosidase using an ultrasound-assisted technique. Atomization of the enzyme solution did not change its activity. The amount of surface-attached beta-galactosidase was measured through its enzymatic reaction product D-galactose using a standardized method. A near-linear increase was obtained in the thickness of the enzyme coat as the treatment proceeded. Interestingly, lactose, which is a substrate for beta-galactosidase, did not undergo enzymatic degradation during processing and remained unchanged for at least 1 month. Stability of protein-coated lactose was due to the absence of water within the powder, as it was dry after the treatment procedure. In conclusion, we were able to attach the polypeptide to the core particles and determine precisely the coating efficiency of the surface-treated powder using a simple approach.

Spatiotemporal regulation of dynamic cell microenvironment signals based on an azobenzene photoswitch.

Dynamic biochemical and biophysical signals of cellular matrix define and regulate tissue-specific cell functions and fate. To recapitulate this complex environment in vitro, biomaterials based on structural- or degradation-tunable polymers have emerged as powerful platforms for regulating the "on-demand" cell-material dynamic interplay. As one of the most prevalent photoswitch molecules, the photoisomerization of azobenzene demonstrates a unique advantage in the construction of dynamic substrates. Moreover, the development of azobenzene-containing biomaterials is particularly helpful in elucidating cells that adapt to a dynamic microenvironment or integrate spatiotemporal variations of signals. Herein, this minireview, places emphasis on the research progress of azobenzene photoswitches in the dynamic regulation of matrix signals. Some techniques and material design methods have been discussed to provide some theoretical guidance for the rational and efficient design of azopolymer-based material platforms. In addition, considering that the UV-light response of traditional azobenzene photoswitches is not conducive to biological applications, we have summarized the recent approaches to red-shifting the light wavelength for azobenzene activation.