| 1. |
- Arsana, Komang G.Y., et al.
(author)
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Laboratory Liquid-Jet X-ray Microscopy and X-ray Fluorescence Imaging for Biomedical Applications
- 2024
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In: International Journal of Molecular Sciences. - : MDPI AG. - 1661-6596 .- 1422-0067. ; 25:2
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Journal article (peer-reviewed)abstract
- Diffraction-limited resolution and low penetration depth are fundamental constraints in optical microscopy and in vivo imaging. Recently, liquid-jet X-ray technology has enabled the generation of X-rays with high-power intensities in laboratory settings. By allowing the observation of cellular processes in their natural state, liquid-jet soft X-ray microscopy (SXM) can provide morphological information on living cells without staining. Furthermore, X-ray fluorescence imaging (XFI) permits the tracking of contrast agents in vivo with high elemental specificity, going beyond attenuation contrast. In this study, we established a methodology to investigate nanoparticle (NP) interactions in vitro and in vivo, solely based on X-ray imaging. We employed soft (0.5 keV) and hard (24 keV) X-rays for cellular studies and preclinical evaluations, respectively. Our results demonstrated the possibility of localizing NPs in the intracellular environment via SXM and evaluating their biodistribution with in vivo multiplexed XFI. We envisage that laboratory liquid-jet X-ray technology will significantly contribute to advancing our understanding of biological systems in the field of nanomedical research.
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| 2. |
- Horneck, Gerda, et al.
(author)
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AstRoMap European Astrobiology Roadmap
- 2016
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In: Astrobiology. - : Mary Ann Liebert. - 1531-1074 .- 1557-8070. ; 16:3, s. 201-243
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Research review (peer-reviewed)abstract
- The European AstRoMap project (supported by the European Commission Seventh Framework Programme) surveyed the state of the art of astrobiology in Europe and beyond and produced the first European roadmap for astrobiology research. In the context of this roadmap, astrobiology is understood as the study of the origin, evolution, and distribution of life in the context of cosmic evolution; this includes habitability in the Solar System and beyond. The AstRoMap Roadmap identifies five research topics, specifies several key scientific objectives for each topic, and suggests ways to achieve all the objectives. The five AstRoMap Research Topics are• Research Topic 1: Origin and Evolution of Planetary Systems• Research Topic 2: Origins of Organic Compounds in Space• Research Topic 3: Rock-Water-Carbon Interactions, Organic Synthesis on Earth, and Steps to Life• Research Topic 4: Life and Habitability• Research Topic 5: Biosignatures as Facilitating Life DetectionIt is strongly recommended that steps be taken towards the definition and implementation of a European Astrobiology Platform (or Institute) to streamline and optimize the scientific return by using a coordinated infrastructure and funding system.
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| 3. |
- Kilic, Nuzhet I., et al.
(author)
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Two-Photon Polymerization Printing with High Metal Nanoparticle Loading
- 2023
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In: ACS Applied Materials and Interfaces. - : American Chemical Society (ACS). - 1944-8244 .- 1944-8252. ; 15:42, s. 49794-49804
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Journal article (peer-reviewed)abstract
- Two-photon polymerization (2PP) is an efficient technique to achieve high-resolution, three-dimensional (3D)-printed complex structures. However, it is restricted to photocurable monomer combinations, thus presenting constraints when aiming at attaining functionally active resist formulations and structures. In this context, metal nanoparticle (NP) integration as an additive can enable functionality and pave the way to more dedicated applications. Challenges lay on the maximum NP concentrations that can be incorporated into photocurable resist formulations due to the laser-triggered interactions, which primarily originate from laser scattering and absorption, as well as the limited dispersibility threshold. In this study, we propose an approach to address these two constraints by integrating metallic Rh NPs formed ex situ, purposely designed for this scope. The absence of surface plasmon resonance (SPR) within the visible and near-infrared spectra, coupled with the limited absorption value measured at the laser operating wavelength (780 nm), significantly limits the laser-induced interactions. Moreover, the dispersibility threshold is increased by engineering the NP surface to be compatible with the photocurable resin, permitting us to achieve concentrations of up to 2 wt %, which, to our knowledge, is significantly higher than the previously reported limit (or threshold) for embedded metal NPs. Another distinctive advantage of employing Rh NPs is their role as promising contrast agents for X-ray fluorescence (XRF) bioimaging. We demonstrated the presence of Rh NPs within the whole 2PP-printed structure and emphasized the potential use of NP-loaded 3D-printed nanostructures for medical devices.
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| 4. |
- Li, Yuyang, et al.
(author)
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Synthesis, Physicochemical Characterization, and Cytotoxicity Assessment of Rh Nanoparticles with Different Morphologies-as Potential XFCT Nanoprobes
- 2020
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In: Nanomaterials. - : MDPI AG. - 2079-4991. ; 10:11, s. 2129-2129
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Journal article (peer-reviewed)abstract
- Morphologically controllable synthesis of Rh nanoparticles (NPs) was achieved by the use of additives during polyol synthesis. The effect of salts and surfactant additives including PVP, sodium acetate, sodiumcitrate, CTAB,CTAC,andpotassiumbromideonRhNPsmorphologywasinvestigated. When PVP was used as the only additive, trigonal NPs were obtained. Additives containing Br− ions (CTAB and KBr) resulted in NPs with a cubic morphology, while those with carboxyl groups (sodium citrate and acetate) formed spheroid NPs. The use of Cl− ions (CTAC) resulted in a mixture of polygon morphologies. Cytotoxicity of these NPs was evaluated on macrophages and ovarian cancer cell lines. Membrane integrity and cellular activity are both influenced to a similar extent, for both the cell lines, with respect to the morphology of Rh NPs. The cells exposed to trigonal Rh NPs showed the highest viability, among the NP series. Particles with a mixed polygon morphology had the highest cytotoxic impact, followed by cubic and spherical NPs. The Rh NPs were further demonstrated as contrast agents for X-ray fluorescence computed tomography (XFCT) in a small-animal imaging setting. This work provides a detailed route for the synthesis, morphology control, and characterization of Rh NPs as viable contrast agents for XFCT bio-imaging.
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| 5. |
- Saladino, Giovanni, et al.
(author)
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Carbon Quantum Dots Conjugated Rhodium Nanoparticles as Hybrid Multimodal Contrast Agents
- 2021
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In: Nanomaterials. - : MDPI AG. - 2079-4991. ; 11:9
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Journal article (peer-reviewed)abstract
- Nanoparticle (NP)-based contrast agents enabling different imaging modalities are sought for non-invasive bio-diagnostics. A hybrid material, combining optical and X-ray fluorescence is presented as a bioimaging contrast agent. Core NPs based on metallic rhodium (Rh) have been demonstrated to be potential X-ray Fluorescence Computed Tomography (XFCT) contrast agents. Microwave-assisted hydrothermal method is used for NP synthesis, yielding large-scale NPs within a significantly short reaction time. Rh NP synthesis is performed by using a custom designed sugar ligand (LODAN), constituting a strong reducing agent in aqueous solution, which yields NPs with primary amines as surface functional groups. The amino groups on Rh NPs are used to directly conjugate excitation-independent nitrogen-doped carbon quantum dots (CQDs), which are synthesized through citrate pyrolysis in ammonia solution. CQDs provided the Rh NPs with optical fluorescence properties and improved their biocompatibility, as demonstrated in vitro by Real-Time Cell Analysis (RTCA) on a macrophage cell line (RAW 264.7). The multimodal characteristics of the hybrid NPs are confirmed with confocal microscopy, and X-ray Fluorescence (XRF) phantom experiments.
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| 6. |
- Saladino, Giovanni, et al.
(author)
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Click chemical assembly and validation of bio-functionalized superparamagnetic hybrid microspheres
- 2020
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In: Applied Nanoscience. - : Springer Nature. - 2190-5509 .- 2190-5517.
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Journal article (peer-reviewed)abstract
- Surface derivatized magnetic nanoparticles have been commonly used for magnetic separation. Facile mechanisms are needed to be developed for the design of bio-functionalized magnetic hybrid materials, where the surfaces can be re-generated for the re-use of the developed platforms. Superparamagnetic iron oxide nanoparticles with a diameter below 10 nm were synthesized via a novel microwave-assisted hydrothermal method in the presence of citrate ions, which allowed to obtain uniform and negatively charged nanoparticles. These were then coupled with Poly-l-lysine (PLL), forming micrometer-sized self-assembled spherical entities. Cross-linking the PLL within these microspheres with glutaraldehyde stabilized them chemically and mechanically. The active bio-functionality was introduced by a protein grafting methodology, using m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester (SMBS). The Moringa oleifera Coagulant Protein (MOCP) from a seed extract was employed for its characteristic coagulation activity. The performance of the MOCP functionalized microspheres was evaluated as a function of turbidity removal of problematic colloidal clay from water via magnetic separation, resulting in over 80% of activity within 15 min. Surface of these hybrid materials can be re-generated by treatment with alcohol, allowing their easy magnetic separation and re-use. The rapid and strong response with tunable magnetic property makes these hybrid microspheres a powerful tool for many potential applications, due to the general applicability of the developed methodology.
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| 7. |
- Saladino, Giovanni, et al.
(author)
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Iterative Nanoparticle Bioengineering Enabled by X-Ray Fluorescence Imaging
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Other publication (other academic/artistic)abstract
- Nanoparticles (NPs) are currently developed for drug delivery and molecular imaging. However, they often get intercepted before reaching their target, leading to low targeting efficacy and signal-to-noise ratio. They tend to accumulate in organs like lungs, liver, kidneys, and spleen. The remedy is to iteratively engineerNP surface properties and administration strategies, presently a time-consuming process that includes dissection at different time points. To improve this, we propose a rapid iterative approach using whole-animal X-ray fluorescence (XRF) imaging to systematically evaluate NP distribution in vivo. We applied this method to molybdenum-based NPs and clodronate liposomes for tumor targeting with transient macrophage depletion, leading to reduced accumulations in lungs and liver and eventual tumor detection.XRF computed tomography (XFCT) provided 3D insight into NP distribution within the tumor. We validated the results using a multiscale imaging approach with dye-doped NPs and gene expression analysis for nanotoxicological profiling. XRF imaging holds potential for advancing therapeutics and diagnostics in preclinical pharmacokinetic studies.
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| 8. |
- Saladino, Giovanni Marco, et al.
(author)
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A versatile strategy to synthesize sugar ligand coated superparamagnetic iron oxide nanoparticles and investigation of their antibacterial activity
- 2021
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In: Colloids and Surfaces A. - : Elsevier BV. - 0927-7757 .- 1873-4359. ; 613
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Journal article (peer-reviewed)abstract
- For the time being, a great attention has been given to the search of green and reusable materials with antibacterial properties. The present research focused on the design and synthesis of hybrid structures constituting superparamagnetic iron oxide nanoparticles (SPIONs) coated with sugar ligands (SL), synthesized using a green and efficient microwave (MW)-assisted hydrothermal synthesis. The sugar ligands were selectively engineered to obtain antibacterial characteristics towards multi-drug resistant bacterial strains, which are among the most problematic bacterial species in antibiotic development efforts. The superparamagnetic behavior was obtained by synthesizing core iron oxide nanoparticles with a diameter below twenty nm. The MW-assisted hydrothermal method yielded a uniform coating of SPIONs with several sugar ligands, granting strongly negative-charged surfaces, which have eventually contributed to their bactericidal activity. The research work allowed to get insights into the magnetic properties of the sugar ligand coated SPIONs, as well as on morphological and functional characteristics of the hybrid nanoparticles, by employing both spectroscopy and imaging techniques, such as FT-IR, Scanning/Transmission Electron Microscopy (S/TEM). Detailed characterizations of the nanoparticles' charge, using zeta potential analysis helped to identify the highly charged hybrids for antibacterial applications. Furthermore, studies on the bactericidal properties of selected SL-SPION hybrids highlighted a high selectivity towards both gram-negative and gram-positive bacteria along with improving bactericidal activity of streptomycin/penicillin mixture. Detailed studies done on Pseudomonas aeruginosa revealed that the SPIONs selectively downregulated the virulence factor pyoverdine and altered bacterial morphology depending on the SL chemistry. The synthesized materials with antibacterial activity pave the way for an effective path towards the design and development of nanostructures and coatings against antibiotic-resistant bacterial species.
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| 9. |
- Saladino, Giovanni Marco, et al.
(author)
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Functional Coatings for X-ray Fluorescent Nanoparticles
- 2022
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In: Proceedings of the 6th International Conference on Theoretical and Applied Nanoscience and Nanotechnology, TANN 2022. - : Avestia Publishing.
-
Conference paper (peer-reviewed)abstract
- In recent years, the design and synthesis of bio-compatible coatings leading to hybrid nanoparticles (NPs) as the contrast agents have gained substantial relevance. Furthermore, the addition of several functionalities for bio-imaging applications represents a key step for non-invasive bio-diagnostics. In this context, we design and utilize hybrid nanostructures for X-ray fluorescence computed tomography (XFCT). The combination of a ceramic or metallic core–based on MoO2, Rh or Ru–with a protective shell allows the generation of bio-compatible nanohybrids for dual mode bio-imaging, where the core NPs constitute the X-ray fluorescence (XRF) contrast agents [1]–[3]. Core NPs are synthesized via polyol, hydrothermal or microwave-assisted hydrothermal methods, yielding uniform shape and high dispersibility in aqueous media. Different approaches have been pursued for the fabrication of a bio-compatible shell coating. A modified sol-gel based silica coating process, doped with a commercial fluorophore (Cy5.5), was developed and shown to be applicable to both ceramic and metallic NPs [4], forming core-shell NPs with both optical and X-ray fluorescence properties. Alternatively, carbon quantum dots (CQDs) were synthesized via citrate pyrolysis using microwave-assisted hydrothermal method, exhibiting uniform size distribution (1.6±0.4 nm) and excitation-independent emission (440 nm). Conjugation of these CQDs, via cross-linking, with Rh NPs led to excitation-independent hybrid NPs, with a red-shifted emission wavelength (520 nm), attributed to the reduction of pyrrolic nitrogen on CQDs [5]. These hybrid NPs exhibit improved in vitro biocompatibility in comparison with bare XRF contrast agents. Furthermore, the optical fluorescence–provided by Cy5.5 or CQDs–allows the localization of the NPs in the intracellular environment while the XRF signal from the core NPs is utilized for XFCT, in small animals, leading to both a microscopic and macroscopic bio-imaging contrast agent.
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| 10. |
- Saladino, Giovanni Marco, et al.
(author)
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Iterative nanoparticle bioengineering enabled by x-ray fluorescence imaging
- 2024
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In: Science Advances. - : American Association for the Advancement of Science (AAAS). - 2375-2548. ; 10:12
-
Journal article (peer-reviewed)abstract
- Nanoparticles (NPs) are currently developed for drug delivery and molecular imaging. However, they often get intercepted before reaching their target, leading to low targeting efficacy and signal-to-noise ratio. They tend to accumulate in organs like lungs, liver, kidneys, and spleen. The remedy is to iteratively engineer NP surface properties and administration strategies, presently a time-consuming process that includes organ dissection at different time points. To improve this, we propose a rapid iterative approach using whole-animal x-ray fluorescence (XRF) imaging to systematically evaluate NP distribution in vivo. We applied this method to molybdenum-based NPs and clodronate liposomes for tumor targeting with transient macrophage depletion, leading to reduced accumulations in lungs and liver and eventual tumor detection. XRF computed tomography (XFCT) provided 3D insight into NP distribution within the tumor. We validated the results using a multiscale imaging approach with dye-doped NPs and gene expression analysis for nanotoxicological profiling. XRF imaging holds potential for advancing therapeutics and diagnostics in preclinical pharmacokinetic studies.
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