TY  - JOUR
AB  - Nanoparticles are widely used in biomedical applications due to their small size, large surface area, and unique physicochemical properties. These characteristics make them ideal for drug delivery, diagnostic imaging, and therapeutic interventions. Their ability to interact with biological systems at the cellular and molecular levels enables targeted treatments. Understanding the biodistribution of nanoparticles at the cellular level within whole organisms is crucial for assessing their safety and effectiveness; however, proper technologies have been lacking to achieve this. For example, traditional imaging techniques like magnetic resonance imaging (MRI) and computed tomography (CT) often lack the resolution needed, while tissue-section-based methods miss the whole-body systemic view. Recent tissue clearing methods have emerged as a promising solution for 3D visualization of nanoparticles in entire organs, as they enable cellular-level imaging of whole organisms without the need for sectioning. This review explores advancements in diverse tissue clearing techniques and their application in studying nanoparticle biodistribution, providing insights crucial for the development of nanoparticle-based therapies.
AU  - Rong, Z.
AU  - Ertürk, A.
AU  - Tang, Y.*
AU  - Mai, H.J.*
C1  - 73282
C2  - 56979
CY  - Postfach 101161, 69451 Weinheim, Germany
TI  - Tissue clearing and its application in nanoparticle development.
JO  - Small
PB  - Wiley-v C H Verlag Gmbh
PY  - 2025
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system offers great opportunities for the treatment of numerous diseases by precise modification of the genome. The functional unit of the system is represented by Cas9/sgRNA ribonucleoproteins (RNP), which mediate sequence-specific cleavage of DNA. For therapeutic applications, efficient and cell-specific transport into target cells is essential. Here, Cas9 RNP nanocarriers are described, which are based on lipid-modified oligoamino amides and folic acid (FolA)-PEG to realize receptor-mediated uptake and gene editing in cancer cells. In vitro studies confirm strongly enhanced potency of receptor-mediated delivery, and the nanocarriers enable efficient knockout of GFP and two immune checkpoint genes, PD-L1 and PVR, at low nanomolar concentrations. Compared with non-targeted nanoparticles, FolA-modified nanocarriers achieve substantially higher gene editing including dual PD-L1/PVR gene disruption after injection into CT26 tumors in vivo. In the syngeneic mouse model, dual disruption of PD-L1 and PVR leads to CD8+ T cell recruitment and distinct CT26 tumor growth inhibition, clearly superior to the individual knockouts alone. The reported Cas9 RNP nanocarriers represent a versatile platform for potent and receptor-specific gene editing. In addition, the study demonstrates a promising strategy for cancer immunotherapy by permanent and combined immune checkpoint disruption.
AU  - Lin, Y.*
AU  - Wilk, U.*
AU  - Pöhmerer, J.*
AU  - Hörterer, E.*
AU  - Höhn, M.*
AU  - Luo, X.*
AU  - Mai, H.
AU  - Wagner, E.*
AU  - Lächelt, U.*
C1  - 66871
C2  - 53338
CY  - Postfach 101161, 69451 Weinheim, Germany
TI  - Folate receptor-mediated delivery of Cas9 RNP for enhanced immune checkpoint disruption in cancer cells.
JO  - Small
VL  - 19
IS  - 2
PB  - Wiley-v C H Verlag Gmbh
PY  - 2022
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - Inhaled nanoparticles constitute a potential health hazard due to their size-dependent lung deposition and large surface to mass ratio. Exposure to high levels contributes to the risk of developing respiratory and cardiovascular diseases, as well as of lung cancer. Particle-induced acute phase response may be an important mechanism of action of particle-induced cardiovascular disease. Here, the authors review new important scientific evidence showing causal relationships between inhalation of particle and nanomaterials, induction of acute phase response, and risk of cardiovascular disease. Particle-induced acute phase response provides a means for risk assessment of particle-induced cardiovascular disease and underscores cardiovascular disease as an occupational disease.
AU  - Hadrup, N.*
AU  - Zhernovkov, V.*
AU  - Jacobsen, N.R.*
AU  - Voss, C.
AU  - Strunz, M.
AU  - Schiller, H. B.
AU  - Halappanavar, S.*
AU  - Poulsen, S.S.*
AU  - Kholodenko, B.*
AU  - Stöger, T.
AU  - Saber, A.T.*
AU  - Vogel, U.*
C1  - 58762
C2  - 48308
CY  - Postfach 101161, 69451 Weinheim, Germany
TI  - Acute phase response as a biological mechanism-of-action of (nano)particle-induced cardiovascular disease.
JO  - Small
VL  - 16
IS  - 21
PB  - Wiley-v C H Verlag Gmbh
PY  - 2020
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - Here, amorphous silica nanoparticles (NPs), one of the most abundant nanomaterials, are used as an example to illustrate the utmost importance of surface coverage by functional groups which critically determines biocompatibility. Silica NPs are functionalized with increasing amounts of amino groups, and the number of surface exposed groups is quantified and characterized by detailed NMR and fluorescamine binding studies. Subsequent biocompatibility studies in the absence of serum demonstrate that, irrespective of surface modification, both plain and amine-modified silica NPs trigger cell death in RAW 264.7 macrophages. The in vitro results can be confirmed in vivo and are predictive for the inflammatory potential in murine lungs. In the presence of serum proteins, on the other hand, a replacement of only 10% of surface-active silanol groups by amines is sufficient to suppress cytotoxicity, emphasizing the relevance of exposure conditions. Mechanistic investigations identify a key role of lysosomal injury for cytotoxicity only in the presence, but not in the absence, of serum proteins. In conclusion, this work shows the critical need to rigorously characterize the surface coverage of NPs by their constituent functional groups, as well as the impact of serum, to reliably establish quantitative nanostructure activity relationships and develop safe nanomaterials.
AU  - Hsiao, I.L.*
AU  - Fritsch-Decker, S.*
AU  - Leidner, A.*
AU  - Al-Rawi, M.*
AU  - Hug, V.*
AU  - Diabaté, S.*
AU  - Grage, S.L.*
AU  - Meffert, M.*
AU  - Stöger, T.
AU  - Gerthsen, D.*
AU  - Ulrich, A.S.*
AU  - Niemeyer, C.M.*
AU  - Weiss, C.*
C1  - 55448
C2  - 46364
CY  - Postfach 101161, 69451 Weinheim, Germany
TI  - Biocompatibility of amine-functionalized silica nanoparticles: The role of surface coverage.
JO  - Small
VL  - 15
IS  - 10
PB  - Wiley-v C H Verlag Gmbh
PY  - 2019
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - Targeted delivery of nanomedicine/nanoparticles (NM/NPs) to the site of disease (e.g., the tumor or lung injury) is of vital importance for improved therapeutic efficacy. Multimodal imaging platforms provide powerful tools for monitoring delivery and tissue distribution of drugs and NM/NPs. This study introduces a preclinical imaging platform combining X-ray (two modes) and fluorescence imaging (three modes) techniques for time-resolved in vivo and spatially resolved ex vivo visualization of mouse lungs during pulmonary NP delivery. Liquid mixtures of iodine (contrast agent for X-ray) and/or (nano)particles (X-ray absorbing and/or fluorescent) are delivered to different regions of the lung via intratracheal instillation, nasal aspiration, and ventilator-assisted aerosol inhalation. It is demonstrated that in vivo propagation-based phase-contrast X-ray imaging elucidates the dynamic process of pulmonary NP delivery, while ex vivo fluorescence imaging (e.g., tissue-cleared light sheet fluorescence microscopy) reveals the quantitative 3D drug/particle distribution throughout the entire lung with cellular resolution. The novel and complementary information from this imaging platform unveils the dynamics and mechanisms of pulmonary NM/NP delivery and deposition for each of the delivery routes, which provides guidance on optimizing pulmonary delivery techniques and novel-designed NM for targeting and efficacy.
AU  - Yang, L.
AU  - Gradl, R.*
AU  - Dierolf, M.*
AU  - Möller, W.
AU  - Kutschke, D.
AU  - Feuchtinger, A.
AU  - Hehn, L.*
AU  - Donnelley, M.*
AU  - Günther, B.*
AU  - Achterhold, K.*
AU  - Walch, A.K.
AU  - Stöger, T.
AU  - Razansky, D.
AU  - Pfeiffer, F.*
AU  - Morgan, K.S.*
AU  - Schmid, O.
C1  - 57167
C2  - 47577
CY  - Postfach 101161, 69451 Weinheim, Germany
TI  - Multimodal precision imaging of pulmonary nanoparticle delivery in mice: Dynamics of application, spatial distribution, and dosimetry.
JO  - Small
VL  - 15
IS  - 59
PB  - Wiley-v C H Verlag Gmbh
PY  - 2019
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - Ambient inhalable particulate matter (PM) is a serious health concern worldwide, but especially so in China where high PM concentrations affect huge populations. Atmospheric processes and emission sources cause spatial and temporal variations in PM concentration and chemical composition, but their influence on the toxicological characteristics of PM are still inadequately understood.In this study, we report an extensive chemical and toxicological characterization of size-segregated urban air inhalable PM collected in August and October 2013 from Nanjing, and assess the effects of atmospheric processes and likely emission sources. A549 human alveolar epithelial cells were exposed to day- and nighttime PM samples (25, 75, 150, 200, 300 mu g/ml) followed by analyses of cytotoxicity, genotoxicity, cell cycle, and inflammatory response.PM10-2.5 and PM0.2 caused the greatest toxicological responses for different endpoints, illustrating that particles with differing size and chemical composition activate distinct toxicological pathways in A549 cells. PM10-2.5 displayed the greatest oxidative stress and genotoxic responses; both were higher for the August samples compared with October. In contrast, PM0.2 and PM2.5-1.0 samples displayed high cytotoxicity and substantially disrupted cell cycle; August samples were more cytotoxic whereas October samples displayed higher cell cycle disruption. Several components associated with combustion, traffic, and industrial emissions displayed strong correlations with these toxicological responses. The lower responses for PM1.0-0.2 compared to PM0.2 and PM2.5-1.0 indicate diminished toxicological effects likely due to aerosol aging and lower proportion of fresh emission particles rich in highly reactive chemical components in the PM1.0-0.2 fraction.Different emission sources and atmospheric processes caused variations in the chemical composition and toxicological responses between PM fractions, sampling campaigns, and day and night. The results indicate different toxicological pathways for coarse-mode particles compared to the smaller particle fractions with typically higher content of combustion-derived components. The variable responses inside PM fractions demonstrate that differences in chemical composition influence the induced toxicological responses.
AU  - Neuschmelting, V.
AU  - Harmsen, S.*
AU  - Bézière, N.
AU  - Lockau, H.*
AU  - Hsu, H.T.*
AU  - Huang, R.C.*
AU  - Razansky, D.
AU  - Ntziachristos, V.
AU  - Kircher, M.F.*
C1  - 53715
C2  - 44923
CY  - Po Box 211, 1000 Ae Amsterdam, Netherlands
TI  - Dual-modality surface-enhanced resonance raman scattering and multispectral optoacoustic tomography nanoparticle approach for brain tumor delineation.
JO  - Small
VL  - 14
IS  - 23
PB  - Elsevier Science Bv
PY  - 2018
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - Nanotechnology holds great promise for a plethora of potential applications. The interaction of engineered nanomaterials with living cells, tissues, and organisms is, however, only partly understood. Microscopic investigations of nano-bio interactions are mostly performed with a few model nanoparticles (NPs) which are easy to visualize, such as fluorescent quantum dots. Here the possibility to visualize nonfluorescent NPs with multiphoton excitation is investigated. Signals from silver (Ag), titanium dioxide (TiO2), and silica (SiO2) NPs in nonbiological environments are characterized to determine signal dependency on excitation wavelength and intensity as well as their signal stability over time. Ag NPs generate plasmon-induced luminescence decaying over time. TiO2 NPs induce photoluminescent signals of variable intensities and in addition strong third harmonic generation (THG). Optimal settings for microscopic detection are determined and then applied for visualization of these two particle types in living cells, in murine muscle tissue, and in the murine blood stream. Silica NPs produce a THG signal, but in living cells it cannot be discriminated sufficiently from endogenous cellular structures. It is concluded that multiphoton excitation is a viable option for studies of nano-bio interactions not only for fluorescent but also for some types of nonfluorescent NPs.
AU  - Dietzel, S.*
AU  - Hermann, S.
AU  - Kugel, Y.*
AU  - Sellner, S.*
AU  - Uhl, B.*
AU  - Hirn, S.*
AU  - Krombach, F.*
AU  - Rehberg, M.*
C1  - 48546
C2  - 41169
CY  - Weinheim
SP  - 3245-3257
TI  - Multiphoton microscopy of nonfluorescent nanoparticles in vitro and in vivo.
JO  - Small
VL  - 12
IS  - 24
PB  - Wiley-v C H Verlag Gmbh
PY  - 2016
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - On page 68, PEGylated gold nanoprisms are designed and prepared by J. M. de la Fuente, D. Cui, and co-workers, with the aim to study the feasibility of using them as a novel contrast agent for the hybrid technique of optoacoustic imaging. The nanoprisms are imaged at different scales using different imaging modalities. They are confirmed as biocompatible, and selected colon cancer HT-29 cells are used as research targets. Shown here is an in silico electron tomographic reconstruction of such gold nanostructures, which show promise for application in biomedical imaging, drug delivery, and photothermal therapy.
AU  - Bao, C.*
AU  - Bézière, N.
AU  - del Pino, P.*
AU  - Pelaz, B.*
AU  - Estrada, G.
AU  - Tian, F.
AU  - Ntziachristos, V.
AU  - de la Fuente, J.M.*
AU  - Cui, D.*
C1  - 11820
C2  - 30823
SP  - 67
TI  - Nanoprisms: Gold nanoprisms as optoacoustic signal nanoamplifiers for in vivo bioimaging of gastrointestinal cancers.
JO  - Small
VL  - 9
IS  - 1
PB  - Wiley-Blackwell
PY  - 2013
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - Early detection of cancer greatly increases the chances of a simpler and more effective treatment. Traditional imaging techniques are often limited by shallow penetration, low sensitivity, low specificity, poor spatial resolution or the use of ionizing radiation. Hybrid modalities, like optoacoustic imaging, an emerging molecular imaging modality, contribute to improving most of these limitations. However, this imaging method is hindered by relatively low signal contrast. Here, gold nanoprisms (AuNPrs) are used as signal amplifiers in multispectral optoacoustic tomography (MSOT) to visualize gastrointestinal cancer. PEGylated AuNPrs are successfully internalized by HT-29 gastrointestinal cancer cells in vitro. Moreover, the particles show good biocompatibility and exhibit a surface plasmon band centered at 830 nm, a suitable wavelength for optoacoustic imaging purposes. These findings extend well to an in vivo setting, in which mice are injected with PEGylated AuNPrs in order to visualize tumor angiogenesis in gastrointestinal cancer cells. Overall, both our in vitro and in vivo results show that PEGylated AuNPrs have the capacity to penetrate tumors and provide a high-resolution signal amplifier for optoacoustic imaging. The combination of PEGylated AuNPrs and MSOT represents a significant advance for the in vivo imaging of cancers.
AU  - Bao, C.*
AU  - Bézière, N.
AU  - del Pino, P.*
AU  - Pelaz, B.*
AU  - Estrada, G.
AU  - Tian, F.
AU  - Ntziachristos, V.
AU  - de la Fuente, J.M.*
AU  - Cui, D.*
C1  - 11320
C2  - 30610
SP  - 68-74
TI  - Gold nanoprisms as optoacoustic signal nanoamplifiers for in vivo bioimaging of gastrointestinal cancers.
JO  - Small
VL  - 9
IS  - 1
PB  - Wiley-VCH
PY  - 2013
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - Understanding the structure and function of glucose binding proteins (GBP) complexed with single walled carbon nanotubes (SWNTs) is important for the development of applications including fluorescent sensors and nanostructure particle tracking. Herein, circular dichroism (CD), thermal denaturation, photo-absorption spectroscopy and atomic force microscopy are used to study these nanostructures. The protein retains its glucose-binding activity after complexation and is thermally stable below 36 °C. However, the SWNT lowers the midpoint denaturation temperature (Tm) by 5 °C and 4 °C in the absence and presence of 10 mM glucose, respectively. This data highlights that using techniques such as CD and thermal denaturation may be necessary to fully characterize such protein-nanomaterial nanostructures.
AU  - McNicholas, T.P.*
AU  - Yum, K.*
AU  - Ahn, J.H.*
AU  - Mu, B.*
AU  - Plettenburg, O.*
AU  - Gooderman, A.*
AU  - Natesan, S.*
AU  - Strano, M.S.*
C1  - 49405
C2  - 0
SP  - 3510-3516
TI  - Structure and function of glucose binding protein-single walled carbon nanotube complexes.
JO  - Small
VL  - 8
IS  - 22
PY  - 2012
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - This study addresses the cellular uptake and intracellular trafficking of 15-nm gold nanoparticles (NPs), either plain (i.e., stabilized with citrate) or coated with polyethylene glycol (PEG), exposed to human alveolar epithelial cells (A549) at the air-liquid interface for 1, 4, and 24 h. Quantitative analysis by stereology on transmission electron microscopy images reveals a significant, nonrandom intracellular distribution for both NP types. No particles are observed in the nucleus, mitochondria, endoplasmic reticulum, or golgi. The cytosol is not a preferred cellular compartment for both NP types, although significantly more PEG-coated than citrate-stabilized NPs are present there. The preferred particle localizations are vesicles of different sizes (<150, 150-1000, >1000 nm). This is observed for both NP types and indicates a predominant uptake by endocytosis. Subsequent inhibition of caveolin- and clathrin-mediated endocytosis by methyl-beta-cyclodextrin (MbetaCD) results in a significant reduction of intracellular NPs. The inhibition, however, is more pronounced for PEG-coated than citrate-stabilized NPs. The latter are mostly found in larger vesicles; therefore, they are potentially taken up by macropinocytosis, which is not inhibited by MbetaCD. With prolonged exposure times, both NPs are preferentially localized in larger-sized intracellular vesicles such as lysosomes, thus indicating intracellular particle trafficking. This quantitative evaluation reveals that NP surface coatings modulate endocytotic uptake pathways and cellular NP trafficking. Other nonendocytotic entry mechanisms are found to be involved as well, as indicated by localization of a minority of PEG-coated NPs in the cytosol.
AU  - Brandenberger, C.*
AU  - Mühlfeld, C.*
AU  - Ali, Z.
AU  - Lenz, A.-G.
AU  - Schmid, O.
AU  - Parak, W.J.*
AU  - Gehr, P.*
AU  - Rothen-Rutishauser, B.*
C1  - 5179
C2  - 27610
SP  - 1669-1678
TI  - Quantitative evaluation of cellular uptake and trafficking of plain and polyethylene glycol-coated gold nanoparticles.
JO  - Small
VL  - 6
IS  - 15
PB  - Wiley-VCH
PY  - 2010
SN  - 1613-6810
ER  - 

TY  - JOUR
AB  - no Abstract
AU  - Semmler-Behnke, M.
AU  - Kreyling, W.G.
AU  - Lipka, J.*
AU  - Fertsch, S.
AU  - Wenk, A.
AU  - Takenaka, S.
AU  - Schmid, G.*
AU  - Brandau, W.*
C1  - 3189
C2  - 26026
SP  - 2108-2111
TI  - Biodistribution of 1.4- and 18nm Gold Particles in Rats.
JO  - Small
VL  - 4
IS  - 12
PB  - Wiley-VCH
PY  - 2008
SN  - 1613-6810
ER  -