Stanford Advisors

All Publications

  • Iterative nanoparticle bioengineering enabled by x-ray fluorescence imaging. Science advances Saladino, G. M., Brodin, B., Kakadiya, R., Toprak, M. S., Hertz, H. M. 2024; 10 (12): eadl2267


    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.

    View details for DOI 10.1126/sciadv.adl2267

    View details for PubMedID 38517973

    View details for PubMedCentralID PMC11093098

  • Laboratory Liquid-Jet X-ray Microscopy and X-ray Fluorescence Imaging for Biomedical Applications. International journal of molecular sciences Arsana, K. G., Saladino, G. M., Brodin, B., Toprak, M. S., Hertz, H. M. 2024; 25 (2)


    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.

    View details for DOI 10.3390/ijms25020920

    View details for PubMedID 38255992

    View details for PubMedCentralID PMC10815599

  • Two-Photon Polymerization Printing with High Metal Nanoparticle Loading. ACS applied materials & interfaces Kilic, N. I., Saladino, G. M., Johansson, S., Shen, R., McDorman, C., Toprak, M. S., Johansson, S. 2023; 15 (42): 49794-49804


    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.

    View details for DOI 10.1021/acsami.3c10581

    View details for PubMedID 37816209

    View details for PubMedCentralID PMC10614202

  • Organ uptake, toxicity and skin clearance of ruthenium contrast agents monitored in vivo by x-ray fluorescence NANOMEDICINE Vogt, C., Saladino, G. M., Shaker, K., Arsenian-Henriksson, M., Hertz, H. M., Toprak, M. S., Brodin, B. A. 2023


    Aims: To investigate the distribution and toxicity of ruthenium nanoparticles (Ru NPs) injected intravenously in mice. Methods: We synthesized Ru NPs, followed their biodistribution by x-ray fluorescence (XRF) imaging and evaluated organ toxicity by histopathology and gene expression. Results: Ru NPs accumulated, mainly in liver and spleen, where they were phagocyted by tissue macrophages, giving a transient inflammation and oxidative stress response that declined after 2 weeks. Ru NPs gradually accumulated in the skin, which was confirmed by microscopic examination of skin biopsies. Conclusion: Ru NP toxicity in recipient organs is transient. Particles are at least partially excreted by the skin, supporting a role for the skin as a nanoparticle clearing organ.

    View details for DOI 10.2217/nnm-2023-0061

    View details for Web of Science ID 001061631900001

    View details for PubMedID 37665018

  • Magnetoresponsive fluorescent core-shell nanoclusters for biomedical applications. Nanoscale advances Saladino, G. M., Kakadiya, R., Ansari, S. R., Teleki, A., Toprak, M. S. 2023; 5 (5): 1323-1330


    Nowadays, superparamagnetic iron oxide nanoparticles (SPIONs) have a dominant role in many subfields of biomedicine. Owing to their peculiar properties, they can be employed for magnetic separation, drug delivery, diagnostics, and hyperthermia treatments. However, these magnetic nanoparticles (NPs) suffer from low unit magnetization due to size constraints (up to 20-30 nm) to exhibit superparamagnetic character. In this work, we have designed and synthesized superparamagnetic nanoclusters (SP-NCs) with diameters of up to 400 nm with high unit magnetization for enhanced loading capacity. These were synthesized with conventional or microwave-assisted solvothermal methods, in the presence of either of the two biomolecules (citrate or l-lysine) as the capping agent. Primary particle size, SP-NC size, surface chemistry, and the resultant magnetic properties were observed to be significantly influenced by the choice of synthesis route and capping agent. Selected SP-NCs were then coated with a fluorophore-doped silica shell to provide fluorescence properties, in the near-infrared spectrum region, while silica provided high chemical and colloidal stability. Heating efficiency studies were performed under alternating magnetic field on the synthesized SP-NCs, highlighting their potential in hyperthermia treatment. We envision that their enhanced magnetically-active content, fluorescence, magnetic property, and heating efficiency will pave the way to more effective uses in biomedical applications.

    View details for DOI 10.1039/d2na00887d

    View details for PubMedID 36866251

    View details for PubMedCentralID PMC9972542

  • XFCT-MRI hybrid multimodal contrast agents for complementary imaging. Nanoscale Saladino, G. M., Vogt, C., Brodin, B., Shaker, K., Kilic, N. I., Andersson, K., Arsenian-Henriksson, M., Toprak, M. S., Hertz, H. M. 2023; 15 (5): 2214-2222


    Multimodal contrast agents in biomedical imaging enable the collection of more comprehensive diagnostic information. In the present work, we design hybrid ruthenium-decorated superparamagnetic iron oxide nanoparticles (NPs) as the contrast agents for both magnetic resonance imaging (MRI) and X-ray fluorescence computed tomography (XFCT). The NPs are synthesized via a one-pot polyol hot injection route, in diethylene glycol. In vivo preclinical studies demonstrate the possibility of correlative bioimaging with these contrast agents. The complementarity allows accurate localization, provided by the high contrast of the soft tissues in MRI combined with the elemental selectivity of XFCT, leading to NP detection with high specificity and resolution. We envision that this multimodal imaging could find future applications for early tumor diagnosis, improved long-term treatment monitoring, and enhanced radiotherapy planning.

    View details for DOI 10.1039/d2nr05829d

    View details for PubMedID 36625091

  • Carbon Quantum Dots Conjugated Rhodium Nanoparticles as Hybrid Multimodal Contrast Agents. Nanomaterials (Basel, Switzerland) Saladino, G. M., Kilic, N. I., Brodin, B., Hamawandi, B., Yazgan, I., Hertz, H. M., Toprak, M. S. 2021; 11 (9)


    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.

    View details for DOI 10.3390/nano11092165

    View details for PubMedID 34578481

    View details for PubMedCentralID PMC8470909

  • Optical and X-ray Fluorescent Nanoparticles for Dual Mode Bioimaging. ACS nano Saladino, G. M., Vogt, C., Li, Y., Shaker, K., Brodin, B., Svenda, M., Hertz, H. M., Toprak, M. S. 2021; 15 (3): 5077-5085


    Nanoparticle (NP) based contrast agents detectable via different imaging modalities (multimodal properties) provide a promising strategy for noninvasive diagnostics. Core-shell NPs combining optical and X-ray fluorescence properties as bioimaging contrast agents are presented. NPs developed earlier for X-ray fluorescence computed tomography (XFCT), based on ceramic molybdenum oxide (MoO2) and metallic rhodium (Rh) and ruthenium (Ru), are coated with a silica (SiO2) shell, using ethanolamine as the catalyst. The SiO2 coating method introduced here is demonstrated to be applicable to both metallic and ceramic NPs. Furthermore, a fluorophore (Cy5.5 dye) was conjugated to the SiO2 layer, without altering the morphological and size characteristics of the hybrid NPs, rendering them with optical fluorescence properties. The improved biocompatibility of the SiO2 coated NPs without and with Cy5.5 is demonstrated in vitro by Real-Time Cell Analysis (RTCA) on a macrophage cell line (RAW 264.7). The multimodal characteristics of the core-shell NPs are confirmed with confocal microscopy, allowing the intracellular localization of these NPs in vitro to be tracked and studied. In situ XFCT successfully showed the possibility of in vivo multiplexed bioimaging for multitargeting studies with minimum radiation dose. Combined optical and X-ray fluorescence properties empower these NPs as effective macroscopic and microscopic imaging tools.

    View details for DOI 10.1021/acsnano.0c10127

    View details for PubMedID 33587608

    View details for PubMedCentralID PMC8028327

  • A versatile strategy to synthesize sugar ligand coated superparamagnetic iron oxide nanoparticles and investigation of their antibacterial activity COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS Saladino, G., Hamawandi, B., Demir, M., Yazgan, I., Toprak, M. 2021; 613
  • Synthesis, Physicochemical Characterization, and Cytotoxicity Assessment of Rh Nanoparticles with Different Morphologies-as Potential XFCT Nanoprobes. Nanomaterials (Basel, Switzerland) Li, Y., Saladino, G. M., Shaker, K., Svenda, M., Vogt, C., Brodin, B., Hertz, H. M., Toprak, M. S. 2020; 10 (11)


    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, sodium citrate, CTAB, CTAC, and potassium bromide on Rh NPs morphology was investigated. 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.

    View details for DOI 10.3390/nano10112129

    View details for PubMedID 33120889

    View details for PubMedCentralID PMC7692549

  • Click chemical assembly and validation of bio-functionalized superparamagnetic hybrid microspheres APPLIED NANOSCIENCE Saladino, G. M., Hamawandi, B., Vogt, C., Rajarao, G. K., Toprak, M. S. 2020; 10 (6): 1861-1869