Investigation of Liquid Collagen Ink for Three-Dimensional Printing.

Three-dimensional printing provides more versatility in the fabrication of scaffold materials for hard and soft tissue replacement, but a critical component is the ink. The ink solution should be biocompatible, stable, and able to maintain scaffold shape, size, and function once printed. This paper describes the development of a collagen ink that remains in a liquid pre-fibrillized state prior to printing. The liquid stability occurs due to the incorporation of ethylenediaminetetraacetic acid (EDTA) during dialysis of the collagen. Collagen inks were 3D-printed using two different printers. The resulting scaffolds were further processed using two different chemical crosslinkers, 1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride)/N-hydroxysuccinimide (EDC/NHS) and genipin; gold nanoparticles were conjugated to the scaffolds. The 3D-printed scaffolds were characterized to determine their extrudability, stability, amount of AuNP conjugated, and overall biocompatibility via cell culture studies using fibroblast cells and stroma cells. The results demonstrated that the liquid collagen ink was amendable to 3D printing and was able to maintain its 3D shape. The scaffolds could be conjugated with gold nanoparticles and demonstrated enhanced biocompatibility. It was concluded that the liquid collagen ink is a good candidate material for the 3D printing of tissue scaffolds.

Minimizing experimental artefacts in synchrotron-based X-ray analyses of Fe speciation in tissues of rice plants.

Iron (Fe) plays an important role within environmental systems. Synchrotron-based X-ray approaches, including X-ray absorption spectroscopy (XAS), provide powerful tools for in situ analyses of Fe speciation, but beam damage during analysis may alter Fe speciation during its measurement. XAS was used to examine whether experimental conditions affect the analysis of Fe speciation in plant tissues. Even when analyzed in a cryostat at 12 K, it was found that FeIII can rapidly (within 0.5-1 min) photoreduce to FeII, although the magnitude of photoreduction varied depending upon the hydration of the sample, the coordination chemistry of the Fe, as well as other properties. For example, photoreduction of FeIII was considerably higher for aqueous standard compounds than for hydrated plant-root tissues. The use of freeze-dried samples in the cryostat (12 K) markedly reduced the magnitude of this FeIII photoreduction, and there was no evidence that the freeze-drying process itself resulted in experimental artefacts under the current experimental conditions, such as through the oxidation of FeII, although some comparatively small differences were observed when comparing spectra of hydrated and freeze-dried FeII compounds. The results of this study have demonstrated that FeIII photoreduction can occur during X-ray analysis, and provides suitable conditions to preserve Fe speciation to minimize the extent of beam damage when analyzing environmental samples. All studies utilizing XAS are encouraged to include a preliminary experiment to determine if beam damage is occurring, and, where appropriate, to take the necessary steps (such as freeze drying) to overcome these issues.

Comparison of KP1019 and NAMI-A in tumour-mimetic environments.

NAMI-A and KP1019 are Ru(III)-based anti-metastatic and cytotoxic anti-cancer drugs, respectively, and have been proposed to be activated by reduction to Ru(II). The potential reduction of NAMI-A and KP1019 in the hypoxic environment of a tumour model of neuroblastoma was examined. Normoxic, hypoxic and necrotic tumour tissues were modelled by multicellular spheroids of SH-SY5Y human neuroblastoma cells of various diameters (50-800 µm). The variation in spheroid environment was confirmed with pimonidazole staining. Laser-ablation inductively-coupled plasma mass spectrometry showed KP1019 and NAMI-A penetration into the spheroid hypoxic region. XANES showed that the speciation of NAMI-A biotransformation products did not change significantly as hypoxia levels increased. KP1019 metabolites showed a correlation between the degree of spheroid hypoxia and the Ru K-edge energy consistent with either partial reduction of Ru(III) to Ru(II) in tumour microenvironments, increased S/Cl coordination or a reduced fraction of polynuclear Ru species. EXAFS spectroscopy was undertaken in an attempt to distinguish between these scenarios but was inconclusive.

Reduction of U(VI) by Fe(II) during the Fe(II)-accelerated transformation of ferrihydrite.

X-ray absorption spectroscopy has been used to study the reduction of adsorbed U(VI) during the Fe(II)-accelerated transformation of ferrihydrite to goethite. The fate of U(VI) was examined across a variety of pH values and Fe(II) concentrations, with results suggesting that, in all cases, it was reduced over the course of the Fe(III) phase transformation to a U(V) species incorporated in goethite. A positive correlation between U(VI) reduction and ferrihydrite transformation rate constants implies that U(VI) reduction was driven by the production of goethite under the conditions used in these studies. This interpretation was supported by additional experimental evidence that demonstrated the (fast) reduction of U(VI) to U(V) by Fe(II) in the presence of goethite only. Theoretical redox potential calculations clearly indicate that the reduction of U(VI) by Fe(II) in the presence of goethite is thermodynamically favorable. In contrast, reduction of U(VI) by Fe(II) in the presence of ferrihydrite is largely thermodynamically unfavorable within the range of conditions examined in this study.

Effect of solution and solid-phase conditions on the Fe(II)-accelerated transformation of ferrihydrite to lepidocrocite and goethite.

Aqueous ferrous iron (Fe(II)) accelerates the transformation of ferrihydrite into secondary, more crystalline minerals however the factors controlling the rate and, indeed, the underlying mechanism of this transformation process remain unclear. Here, we present the first detailed study of the kinetics of the Fe(II)-accelerated transformation of ferrihydrite to goethite, via lepidocrocite, for a range of pH and Fe(II) concentrations and, from the results obtained, provide insight into the factors controlling the transformation rate and the processes responsible for transformation. A reaction scheme for the Fe(II)-accelerated secondary mineralization of ferrihydrite is developed in which an Fe(II) atom attaches to the ferrihydrite surface where it is immediately oxidized to Fe(III) with the resultant electron transferred, sequentially, to other iron oxyhydroxide Fe(III) atoms before release to solution as Fe(II). This freshly precipitated Fe(III) forms the nuclei for the formation of secondary minerals and also facilitates the ongoing uptake of Fe(II) from solution by creation of fresh surface sites. The concentration of solid-associated Fe(II) and the rate of transport of Fe(II) to the oxyhydroxide surface appear to determine which particular secondary minerals form and their rates of formation. Lepidocrocite growth is enhanced at lower solid-associated Fe(II) concentrations while conditions leading to more rapid uptake of Fe(II) from solution lead to higher goethite growth rates.

Fate of ZnO nanoparticles in soils and cowpea (Vigna unguiculata).

The increasing use of zinc oxide nanoparticles (ZnO-NPs) in various commercial products is prompting detailed investigation regarding the fate of these materials in the environment. There is, however, a lack of information comparing the transformation of ZnO-NPs with soluble Zn(2+) in both soils and plants. Synchrotron-based techniques were used to examine the uptake and transformation of Zn in various tissues of cowpea ( Vigna unguiculata (L.) Walp.) exposed to ZnO-NPs or ZnCl2 following growth in either solution or soil culture. In solution culture, soluble Zn (ZnCl2) was more toxic than the ZnO-NPs, although there was substantial accumulation of ZnO-NPs on the root surface. When grown in soil, however, there was no significant difference in plant growth and accumulation or speciation of Zn between soluble Zn and ZnO-NP treatments, indicating that the added ZnO-NPs underwent rapid dissolution following their entry into the soil. This was confirmed by an incubation experiment with two soils, in which ZnO-NPs could not be detected after incubation for 1 h. The speciation of Zn was similar in shoot tissues for both soluble Zn and ZnO-NPs treatments and no upward translocation of ZnO-NPs from roots to shoots was observed in either solution or soil culture. Under the current experimental conditions, the similarity in uptake and toxicity of Zn from ZnO-NPs and soluble Zn in soils indicates that the ZnO-NPs used in this study did not constitute nanospecific risks.

In situ speciation and distribution of toxic selenium in hydrated roots of cowpea.

The speciation and spatial distribution of selenium (Se) in hydrated plant tissues is not well understood. Using synchrotron-based x-ray absorption spectroscopy and x-ray fluorescence microscopy (two-dimensional scanning [and associated mathematical model] and computed tomography), the speciation and distribution of toxic Se were examined within hydrated roots of cowpea (Vigna unguiculata) exposed to either 20 µM selenite or selenate. Based upon bulk solution concentrations, selenate was 9-fold more toxic to the roots than selenite, most likely due to increased accumulation of organoselenium (e.g. selenomethionine) in selenate-treated roots. Specifically, uptake of selenate (probably by sulfate transporters) occurred at a much higher rate than for selenite (apparently by both passive diffusion and phosphate transporters), with bulk root tissue Se concentrations approximately 18-fold higher in the selenate treatment. Although the proportion of Se converted to organic forms was higher for selenite (100%) than for selenate (26%), the absolute concentration of organoselenium was actually approximately 5-fold higher for selenate-treated roots. In addition, the longitudinal and radial distribution of Se in roots differed markedly: the highest tissue concentrations were in the endodermis and cortex approximately 4 mm or more behind the apex when exposed to selenate but in the meristem (approximately 1 mm from the apex) when exposed to selenite. The examination of the distribution and speciation of Se in hydrated roots provides valuable data in understanding Se uptake, transport, and toxicity.

Structure and cation ordering in spinel-type TcCo2O4. An example of a trivalent technetium oxide.

The structure of TcCo(2)O(4) has been determined using a combination of synchrotron X-ray and neutron powder diffraction methods. It has an inverse spinel structure where the Tc occupies the octahedral sites. Both the refined Tc-O distance and X-ray absorption spectra suggest the Tc is predominantly trivalent.

In situ distribution and speciation of toxic copper, nickel, and zinc in hydrated roots of cowpea.

The phytotoxicity of trace metals is of global concern due to contamination of the landscape by human activities. Using synchrotron-based x-ray fluorescence microscopy and x-ray absorption spectroscopy, the distribution and speciation of copper (Cu), nickel (Ni), and zinc (Zn) was examined in situ using hydrated roots of cowpea (Vigna unguiculata) exposed to 1.5 µm Cu, 5 µm Ni, or 40 µm Zn for 1 to 24 h. After 24 h of exposure, most Cu was bound to polygalacturonic acid of the rhizodermis and outer cortex, suggesting that binding of Cu to walls of cells in the rhizodermis possibly contributes to the toxic effects of Cu. When exposed to Zn, cortical concentrations remained comparatively low with much of the Zn accumulating in the meristematic region and moving into the stele; approximately 60% to 85% of the total Zn stored as Zn phytate within 3 h of exposure. While Ni concentrations were high in both the cortex and meristem, concentrations in the stele were comparatively low. To our knowledge, this is the first report of the in situ distribution and speciation of Cu, Ni, and Zn in hydrated (and fresh) plant tissues, providing valuable information on the potential mechanisms by which they are toxic.