Prophylactic Treatment with Cerium Oxide Nanoparticles Attenuate Hepatic Ischemia Reperfusion Injury in Sprague Dawley Rats.

BACKGROUND: Hepatic ischemia reperfusion is one the main causes for graft failure following transplantation. Although, the molecular events that lead to hepatic failure following ischemia reperfusion (IR) are diverse and complex, previous studies have shown that excessive formation of reactive oxygen species (ROS) are responsible for hepatic IR injury. Cerium oxide (CeO2) nanoparticles have been previously shown to act as an anti-oxidant and anti-inflammatory agent. Here, we evaluated the protective effects of CeO2 nanoparticles on hepatic ischemia reperfusion injury. METHODS: Male Sprague Dawley rats were randomly assigned to one of the four groups: Control, CeO2 nanoparticle only, hepatic ischemia reperfusion (IR) group and hepatic ischemia reperfusion (IR) plus CeO2 nanoparticle group (IR+ CeO2). Partial warm hepatic ischemia was induced in left lateral and median lobes for 1h, followed by 6h of reperfusion. Animals were sacrificed after 6h of reperfusion and blood and tissue samples were collected and processed for various biochemical experiments. RESULTS: Prophylactic treatment with CeO2 nanoparticles (0.5mg/kg i.v (IR+CeO2 group)) 1 hour prior to hepatic ischemia and subsequent reperfusion injury lead to a decrease in serum levels of alanine aminotransaminase and lactate dehydrogenase at 6 hours after reperfusion. These changes were accompanied by significant decrease in hepatocyte necrosis along with reduction in several serum inflammatory markers such as macrophage derived chemokine, macrophage inflammatory protein-2, KC/GRO, myoglobin and plasminogen activator inhibitor-1. However, immunoblotting demonstrated no significant changes in the levels of apoptosis related protein markers such as bax, bcl2 and caspase 3 in IR and IR+ CeO2 groups at 6 hours suggesting necrosis as the main pathway for hepatocyte death. CONCLUSION: Taken together, these data suggest that CeO2 nanoparticles attenuate IR induced cell death and can be used as a prophylactic agent to prevent hepatic injury associated with graft failure.

Efficiency of Nanotube Surface-Treated Dental Implants Loaded with Doxycycline on Growth Reduction of Porphyromonas gingivalis.

PURPOSE: The prevalence of peri-implant infection in patients with dental implants has been shown to range from 28% to 56%. A nanotube-modified implant surface can deliver antibiotics locally and suppress periodontal pathogenic bacterial growth. The aim of this study was to evaluate the deliverability of antibiotics via a nanotube-modified implant. MATERIALS AND METHODS: Dental implants with a nanotube surface were fabricated and loaded with doxycycline. Afterward, each dental implant with a nanotube surface was placed into 2-mL tubes, removed from solution, and placed in a fresh solution daily for 28 days. Experimental samples from 1, 2, 4, 16, 24, and 28 days were used for this evaluation. The concentration of doxycycline was measured using spectrophotometric analysis at 273-nm absorbance. The antibacterial effect of doxycycline was evaluated by supplementing Porphyromonas gingivalis (P gingivalis) growth media with the solution collected from the dental implants at the aforementioned time intervals for a period of 48 hours under anaerobic conditions. A bacterial viability assay was used to evaluate P gingivalis growth at 550-nm absorbance. RESULTS: Doxycycline concentration varied from 0.33 to 1.22 µg/mL from day 1 to day 28, respectively. A bacterial viability assay showed the highest P gingivalis growth at day 1 (2 nm) and the lowest at day 4 (0.17 nm), with a gradual reduction from day 1 to day 4 of approximately 87.5%. The subsequent growth pattern was maintained and slightly increased from baseline in approximately 48.3% from day 1 to day 24. The final P gingivalis growth measured at day 28 was 29.4% less than the baseline growth. CONCLUSION: P gingivalis growth was suppressed in media supplemented with solution collected from dental implants with a nanotube surface loaded with doxycycline during a 28-day time interval.

Thermally oxidized titania nanotubes enhance the corrosion resistance of Ti6Al4V.

The negative impact of in vivo corrosion of metallic biomedical implants remains a complex problem in the medical field. We aimed to determine the effects of electrochemical anodization (60V, 2h) and thermal oxidation (600°C) on the corrosive behavior of Ti-6Al-4V, with serum proteins, at physiological temperature. Anodization produced a mixture of anatase and amorphous TiO2 nanopores and nanotubes, while the annealing process yielded an anatase/rutile mixture of TiO2 nanopores and nanotubes. The surface area was analyzed by the Brunauer-Emmett-Teller method and was estimated to be 3 orders of magnitude higher than that of polished control samples. Corrosion resistance was evaluated on the parameters of open circuit potential, corrosion potential, corrosion current density, passivation current density, polarization resistance and equivalent circuit modeling. Samples both anodized and thermally oxidized exhibited shifts of open circuit potential and corrosion potential in the noble direction, indicating a more stable nanoporous/nanotube layer, as well as lower corrosion current densities and passivation current densities than the smooth control. They also showed increased polarization resistance and diffusion limited charge transfer within the bulk oxide layer. The treatment groups studied can be ordered from greatest corrosion resistance to least as Anodized+Thermally Oxidized > Anodized > Smooth > Thermally Oxidized for the conditions investigated. This study concludes that anodized surface has a potential to prevent long term implant failure due to corrosion in a complex in-vivo environment.

A Novel Investigation of the Formation of Titanium Oxide Nanotubes on Thermally Formed Oxide of Ti-6Al-4V.

Traditionally, titanium oxide (TiO2) nanotubes (TNTs) are anodized on Ti-6Al-4V alloy (Ti-V) surfaces with native TiO2 (amorphous TiO2); subsequent heat treatment of anodized surfaces has been observed to enhance cellular response. As-is bulk Ti-V, however, is often subjected to heat treatment, such as thermal oxidation (TO), to improve its mechanical properties. Thermal oxidation treatment of Ti-V at temperatures greater than 200°C and 400°C initiates the formation of anatase and rutile TiO2, respectively, which can affect TNT formation. This study aims at understanding the TNT formation mechanism on Ti-V surfaces with TO-formed TiO2 compared with that on as-is Ti-V surfaces with native oxide. Thermal oxidation-formed TiO2 can affect TNT formation and surface wettability because TO-formed TiO2 is expected to be part of the TNT structure. Surface characterization was carried out with field emission scanning electron microscopy, energy dispersive x-ray spectroscopy, water contact angle measurements, and white light interferometry. The TNTs were formed on control and 300°C and 600°C TO-treated Ti-V samples, and significant differences in TNT lengths and surface morphology were observed. No difference in elemental composition was found. Thermal oxidation and TO/anodization treatments produced hydrophilic surfaces, while hydrophobic behavior was observed over time (aging) for all samples. Reduced hydrophobic behavior was observed for TO/anodized samples when compared with control, control/anodized, and TO-treated samples. A method for improved surface wettability and TNT morphology is therefore discussed for possible applications in effective osseointegration of dental and orthopedic implants.

Atomic Layer Deposition in Bio-Nanotechnology: A Brief Overview.

Atomic layer deposition (ALD) is a technique increasingly used in nanotechnology and ultrathin film deposition; it is ideal for films in the nanometer and Angstrom length scales. ALD can effectively be used to modify the surface chemistry and functionalization of engineering-related and biologically important surfaces. It can also be used to alter the mechanical, electrical, chemical, and other properties of materials that are increasingly used in biomedical engineering and biological sciences. ALD is a relatively new technique for optimizing materials for use in bio-nanotechnology. Here, after a brief review of the more widely used modes of ALD and a few of its applications in biotechnology, selected results that show the potential of ALD in bio-nanotechnology are presented. ALD seems to be a promising means for tuning the hydrophilicity/hydrophobicity characteristics of biomedical surfaces, forming conformal ultrathin coatings with desirable properties on biomedical substrates with a high aspect ratio, tuning the antibacterial properties of substrate surfaces of interest, and yielding multifunctional biomaterials for medical implants and other devices.

Fabrication of anti-aging TiO2 nanotubes on biomedical Ti alloys.

The primary objective of this study was to fabricate a TiO2 nanotubular surface, which could maintain hydrophilicity over time (resist aging). In order to achieve non-aging hydrophilic surfaces, anodization and annealing conditions were optimized. This is the first study to show that anodization and annealing condition affect the stability of surface hydrophilicity. Our results indicate that maintenance of hydrophilicity of the obtained TiO2 nanotubes was affected by anodization voltage and annealing temperature. Annealing sharply decreased the water contact angle (WCA) of the as-synthesized TiO2 nanotubular surface, which was correlated to improved hydrophilicity. TiO2 nanotubular surfaces are transformed to hydrophilic surfaces after annealing, regardless of annealing and anodization conditions; however, WCA measurements during aging demonstrate that surface hydrophilicity of non-anodized and 20 V anodized samples decreased after only 11 days of aging, while the 60 V anodized samples maintained their hydrophilicity over the same time period. The nanotubes obtained by 60 V anodization followed by 600 °C annealing maintained their hydrophilicity significantly longer than nanotubes which were obtained by 60 V anodization followed by 300 °C annealing.

Novel functionalization of Ti-V alloy and Ti-II using atomic layer deposition for improved surface wettability.

Surface wettability characteristics of commercially pure titanium (CP-Ti/Ti-II) and titanium Grade 5 alloy (Ti-6Al-4V/Ti-V) with 10nm-thick atomic layer deposited (ALD) TiO2 from Tetrakis DiEthyl Amino Titanium and water vapor were studied in conjunction with cleaning steps before and after the ALD treatment. The wettability characteristics of rough Ti-II and Ti-V samples were investigated after each step, that is, as received, after de-ionized (DI) water rinse followed by N2 drying, sonication in methanol, ALD treatment, and post-ALD DI water rinse. Samples without ALD or cleaning treatments were hydrophobic to variable extents, depending on exposure to different environments, surface impurities, roughness, and aging. Surface treatments reported in the literature resulted in hydrophilic/hydrophobic surfaces likely due to organic and/or inorganic impurities. In this study, (i) it is established that it is critically important to probe surface wettability after each substrate treatment; (ii) both Ti-II and Ti-V surfaces are found to become more hydrophilic after each one of the sequential treatments used; and (iii) independently of the initial wettability characteristics of Ti-II and Ti-V surfaces, the aforementioned treatments result in a water contact angle well below 10°, which is an important factor in cellular response. X-ray photoelectron spectroscopy of ALD titania films indicated trace impurities in them. Grazing incidence X-ray diffraction suggested amorphous ALD TiO2 at 200 °C; anatase TiO2 was obtained with as little as 5 min annealing at 600 °C in nitrogen.

Enhancing surface characteristics of Ti-6Al-4V for bio-implants using integrated anodization and thermal oxidation.

Modifications of Ti-6Al-4V surface roughness, wettability and composition are increasingly studied to improve cellular viability on biomedical implants involving Ti-6Al-4V. In this study, it is shown that modification of Ti-6Al-4V samples using anodization (for the formation of titania nanotubes) combined with thermal oxidation (TO) results in superior surface characteristics to those of a smooth, rough, anodized-smooth or anodized-rough surface alone. Surface characterization is performed using water contact angle (WCA) measurements, white-light interferometry, Fourier transform infrared spectroscopy (FTIRS), field emission scanning electron microscopy and grazing incidence X-ray diffraction (GIXRD). WCA measurements before TO indicate that anodized-smooth and anodized-rough samples are super-hydrophilic (WCA less than 5°); WCA of non-anodized smooth and rough surfaces are 57 ± 6° and 86 ± 7°, respectively. After TO at 450 °C for 3 hours, all samples become super-hydrophilic; however, three weeks after TO, smooth and rough surfaces become hydrophobic, while anodized-smooth and anodized-rough surfaces remain hydrophilic. FTIRS and GIXRD data show that the TO of anodized and non-anodized smooth samples results in anatase and rutile TiO2, of which anatase is favorable for cellular attachment. Micro-/nano-scale roughness and TO are discussed in the context of enhanced Ti-6Al-4V surface characteristics for improved cellular response.