Internalization mechanisms of cell-penetrating peptides

• Ivana Ruseska and
• Andreas Zimmer

Beilstein J. Nanotechnol. 2020, 11, 101–123, doi:10.3762/bjnano.11.10

• . However, in 2003, Richard et al. [38] showed that the results obtained might be a misconception due to the use of fixed cells. They postulated that the fixatives could change the intracellular distribution of the peptides. Additionally, it was shown that flow cytometry (a method frequently used for

Bombesin receptor-targeted liposomes for enhanced delivery to lung cancer cells

• Mohammad J. Akbar,
• Pâmela C. Lukasewicz Ferreira,
• Melania Giorgetti,
• Leanne Stokes and
• Christopher J. Morris

Beilstein J. Nanotechnol. 2019, 10, 2553–2562, doi:10.3762/bjnano.10.246

• , quantification of cell-uptake was judged by flow cytometry analysis of A549-GRPR cells exposed to fluorescently tagged target-lipo (FL-Target-lipo) or tagged control-lipo (FL-Control-lipo) formulations. Preliminary studies using FL-Target-lipo including 1 mol % targeting lipid showed marginal increases in

• that GRPR targeting with cystabn peptide increases cell uptake of liposomes, most likely through receptor-mediated uptake. Taken together the flow cytometry and microscopy data demonstrate that, using a fluorescently labelled model liposomal carrier, the relative increase in cell uptake afforded by

• human GRP receptor (3xHA-GRPR pcDNA3.1+ plasmid #GRPR00TN00, UMR cDNA Resource Center, USA) using FuGene 6 and selected using 750 μg/mL G418 over three weeks. Cells were subsequently maintained in 100 μg/mL G418. Flow cytometry A549-GRPR cells were washed with warm PBS and blocked for 5 h in 0.2% BSA in

Small protein sequences can induce cellular uptake of complex nanohybrids

• Jan-Philip Merkl,
• Malak Safi,
• Christian Schmidtke,
• Fadi Aldeek,
• Johannes Ostermann,
• Tatiana Domitrovic,
• Sebastian Gärtner,
• John E. Johnson,
• Horst Weller and
• Hedi Mattoussi

Beilstein J. Nanotechnol. 2019, 10, 2477–2482, doi:10.3762/bjnano.10.238

• the overall hybrids, the QD-to-AuNP molar ratio in the hybrids, the incubation time to 30 min, resulted in significantly lower levels of intracellular QD staining. Flow cytometry measurements showed that under these modified conditions approx. 20% of the cells are labelled with the nanohybrids. In

• culture, epifluorescence z-stack, epifluorescence control experiments, flow cytometry, further instrumentation. Supporting Information File 300: Additional experimental data. Acknowledgements This work was supported by the State Excellence Initiative “Nanotechnology in Medicine” from the Free and

Synthesis and potent cytotoxic activity of a novel diosgenin derivative and its phytosomes against lung cancer cells

• Liang Xu,
• Dekang Xu,
• Ziying Li,
• Yu Gao and
• Haijun Chen

Beilstein J. Nanotechnol. 2019, 10, 1933–1942, doi:10.3762/bjnano.10.189

• late apoptosis in human cervical HeLa cancer cells [26]. In order to explore the death mechanism of P2, A549 and PC9 cells were treated with P2 and Di for 48 h and the proportion of cells undergone apoptosis was counted through Annexin V-FITC/PI staining and followed by flow cytometry analysis. As

• ModFit software (Verity Software House, Topsham, ME) [41]. The cell amounts for each group which are analyzed by the flow cytometry are 10000 events. The representative gating for flow cytometry was shown in Supporting Information File 1, Figure S3. Cell apoptosis For analysis of apoptosis, A549 and PC-9

• dark. Subsequently, the apoptotic stages were quantitatively determined by flow cytometry [42]. Statistical analysis All data shown in this article were expressed as the mean ± SD for at least three separate experiments. Statistical analysis was performed using the Student's t-test. (A) The structure

Engineered superparamagnetic iron oxide nanoparticles (SPIONs) for dual-modality imaging of intracranial glioblastoma via EGFRvIII targeting

• Xianping Liu,
• Chengjuan Du,
• Haichun Li,
• Ting Jiang,
• Zimiao Luo,
• Zhiqing Pang,
• Daoying Geng and
• Jun Zhang

Beilstein J. Nanotechnol. 2019, 10, 1860–1872, doi:10.3762/bjnano.10.181

• -EGFRvIII cells was carried out on cells digested and gathered through centrifugation at 1000 rpm for 4 min. Finally, 0.5 mL of PBS (0.01 M, pH 7.4) was used to suspend the cells and the fluorescence intensity of the cells was ascertained by flow cytometry (BD, USA). U87MG and U87MG-EGFRvIII cells were

• centrifugation was performed at 1000 rpm for 4 min. Finally, 0.5 mL of PBS (0.01 M, pH 7.4) was used to suspend the cells and the fluorescence intensity of the cells was ascertained by flow cytometry (BD, USA). Cellular uptake of PEG-SPIONs The cellular uptake of NPs and PNPs were investigated on U87MG and U87MG

• digestion and suspended in 0.5 mL of PBS (0.01 M, pH 7.4) and analyzed with flow cytometry (BD, USA) at 488 nm. In vivo T2-weighted magnetic resonance imaging (MRI) of intracranial glioblastoma with PEG-SPIONs Intracranial glioblastoma models were established as previously described [40][41]. In brief, 5

Nanoarchitectonics meets cell surface engineering: shape recognition of human cells by halloysite-doped silica cell imprints

• Elvira Rozhina,
• Ilnur Ishmukhametov,
• Svetlana Batasheva,
• Farida Akhatova and
• Rawil Fakhrullin

Beilstein J. Nanotechnol. 2019, 10, 1818–1825, doi:10.3762/bjnano.10.176

• occurs. However, flow cytometry-based cell proliferation monitoring performed with cells stained with 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) dye has confirmed that cells coated with pure silica or silica/halloysite display a similar cell proliferation pattern as intact HeLa cells

• nanotubes. Optical microscopy images demonstrating the cultivation for 24 h of (A, B) substrate-attached intact HeLa cells and (C, D) HeLa cells coated with halloysite-doped silica shells; flow cytometry monitoring of cell division for three days of CFSE-stained (E) intact HeLa cells, (F) silica-coated HeLa

• equipped with a fluorite 100× objective and Dage xL (Dage-MTI) CCD camera. Images were obtained using Exponent 7 software (Dage-MTI). The dark-field images were overlapped with transmission fluorescence images using the image processing software GIMP. Cell proliferative activity was examined by flow

Toxicity and safety study of silver and gold nanoparticles functionalized with cysteine and glutathione

• Barbara Pem,
• Igor M. Pongrac,
• Lea Ulm,
• Ivan Pavičić,
• Valerije Vrček,
• Darija Domazet Jurašin,
• Marija Ljubojević,
• Adela Krivohlavek and
• Ivana Vinković Vrček

Beilstein J. Nanotechnol. 2019, 10, 1802–1817, doi:10.3762/bjnano.10.175

• Ag L−1). The behavior of GSH AuNPs deviates considerably from the others and did not show any significant change in the side scattered light (SSC) ratio even at the highest dose applied, indicating no cell accumulation. The flow cytometry results on NP uptake were confirmed by visualization performed

• apoptotic/dead cells were determined by flow cytometry analysis after staining with CAM and EthD dyes. The doses that caused low or no significant reduction in cell viability (Figure 4a) showed apoptotic processes in a dose-response manner for all tested species (Figure 4b). In accordance with the MTT data

• shaking the plates. The absorbance was recorded at 530 nm using a VictorTM multilabel reader (Perkin Elmer, Massachusetts, USA). The ratio of live, apoptotic and dead cells after treatment with NPs or respective metal salts was determined by flow cytometry experiments using Annexin V and 7

Scavenging of reactive oxygen species by phenolic compound-modified maghemite nanoparticles

• Małgorzata Świętek,
• Yi-Chin Lu,
• Rafał Konefał,
• Liliana P. Ferreira,
• M. Margarida Cruz,
• Yunn-Hwa Ma and
• Daniel Horák

Beilstein J. Nanotechnol. 2019, 10, 1073–1088, doi:10.3762/bjnano.10.108

• -diphenyl-1-picrylhydrazyl (DPPH) assay. Cellular uptake and intracellular antioxidant activity of the particles were evaluated by an iron assay and flow cytometry, respectively, using L-929 and LN-229 cells. Compared to the control, the phenolic modification significantly reduced intracellular reactive

• phenolic compound-modified nanoparticles (100 μg/mL), they were incubated with L-929 and LN-229 cells for 3 h, and 2 mM H2O2 was added for 30 min, followed by staining with CM-H2DCFDA for 1 h. Figure 6 shows the representative flow cytometry results of nanoparticle internalization and the intracellular ROS

• most potent free radical scavenging activity measured by using the DPPH assay, all three phenolic compound-modified nanoparticles reduced intracellular ROS levels in a similar manner as the control, as indicated by flow cytometry. Because intracellular oxidative stress has been shown to be elevated in

The nanoscaled metal-organic framework ICR-2 as a carrier of porphyrins for photodynamic therapy

• Jan Hynek,
• Sebastian Jurík,
• Martina Koncošová,
• Jaroslav Zelenka,
• Ivana Křížová,
• Tomáš Ruml,
• Kaplan Kirakci,
• Ivo Jakubec,
• František Kovanda,
• Kamil Lang and
• Jan Demel

Beilstein J. Nanotechnol. 2018, 9, 2960–2967, doi:10.3762/bjnano.9.275

• Essential Medium (EMEM) without foetal bovine serum to avoid modification of the particle surface properties by nonspecific binding of serum albumin. The cellular uptake of the nanoparticles was quantified by flow-cytometry analysis of porphyrin fluorescence associated with the cells. Figure 6A shows the

• were 405 nm and 488 nm, respectively. During the confocal microscopy, the cells were maintained at 37 °C and 5% CO2 atmosphere. Flow cytometry: The cells were plated onto 6-well plates in full culture medium. The next day, they were treated with indicated amount of nanoparticles for the indicated

• period of time in the fresh medium without serum. Then, the plates were washed with PBS and trypsinized. Uptake of MOFs was measured by flow cytometry analysis with excitation and emission recorded at 405 nm and 655–685 nm, respectively (BD FACSaria III). Structure of ICR-2 viewed along the c axis (left

Enhanced antineoplastic/therapeutic efficacy using 5-fluorouracil-loaded calcium phosphate nanoparticles

• Shanid Mohiyuddin,
• Saba Naqvi and
• Gopinath Packirisamy

Beilstein J. Nanotechnol. 2018, 9, 2499–2515, doi:10.3762/bjnano.9.233

• observed in FE-SEM micrographs. The up-regulated proapoptotic and down-regulated antiapoptotic gene expressions were further confirmed with semiquantitative reverse transcriptase polymerase chain reaction (PCR). The increased intracellular reactive oxygen species (ROS) were quantified via flow cytometry

• oxygen species quantification via flow cytometry The apoptotic induction was experimentally determined by staining techniques, as well as fluorescence and electron microscopy. We further investigated the probable cause of apoptotic induction in cells by CaP@5-FU NPs. It was reported that 5-FU-mediated

• . In its reduced state, CellROX deep red has no or little fluorescence. Upon oxidation by reactive oxygen species, the increased bright red fluorescence emission of CellROX deep red dye can be easily quantified by fluorescence via flow cytometry [46]. The finding was quite interesting whereby 5.1% of

Nanoparticle delivery to metastatic breast cancer cells by nanoengineered mesenchymal stem cells

• Liga Saulite,
• Karlis Pleiko,
• Ineta Popena,
• Dominyka Dapkute,
• Ricardas Rotomskis and
• Una Riekstina

Beilstein J. Nanotechnol. 2018, 9, 321–332, doi:10.3762/bjnano.9.32

• study was to demonstrate that nanoengineered MSCs can serve as a delivery vehicle to target breast cancer cells in a 3D co-culture model. Results Optimal quantum dot labelling conditions in mesenchymal stem cells The flow cytometry data revealed that MSCs incubated in serum-free conditions accumulated

• temperature. Flow cytometry data were acquired on a Guava EasyCyte 8HT flow cytometer and analysed by ExpressPro software (Millipore, MA, USA). Quantum dots Qdot® 655 ITK™ non-targeted carboxyl-coated quantum dots were purchased from Thermo Fisher Scientific, USA. The QDs are composed of a CdSe core and ZnS

• dissociation of the spheroids into a single cell suspension. The cells were washed, stained with CD90 FITC or EpCAM FITC and analysed by flow cytometry. For fluorescence imaging of QD transfer, the cancer cell and MSC mono- or co-culture spheroids were harvested after 24 h of propagation on the polyHEMA

A nanocomplex of C₆₀ fullerene with cisplatin: design, characterization and toxicity

• Svitlana Prylutska,
• Svitlana Politenkova,
• Kateryna Afanasieva,
• Volodymyr Korolovych,
• Kateryna Bogutska,
• Andriy Sivolob,
• Larysa Skivka,
• Maxim Evstigneev,
• Viktor Kostjukov,
• Yuriy Prylutskyy and
• Uwe Ritter

Beilstein J. Nanotechnol. 2017, 8, 1494–1501, doi:10.3762/bjnano.8.149

• compounds. The genotoxicity of С60 fullerene, Cis and their complex was evaluated in vitro with the comet assay using human resting lymphocytes and lymphocytes after blast transformation. The cytotoxicity of the mentioned compounds was estimated by Annexin V/PI double staining followed by flow cytometry

• lymphocytes and reduces the fraction of necrotic cells. Keywords: atomic force microscopy; C60 fullerene; cisplatin; comet assay; computer simulation; dynamic light scattering; flow cytometry; human lymphocytes; toxicity in vitro; Introduction The water-soluble inorganic bi-valent platinum derivative

• with mentioned compounds at the concentration of 0.15 mg/mL for 24 h. After culturing, cells were stained with annexin V (AnnV)/propidium iodide (PI) and analyzed by flow cytometry. Control: cells were incubated without any additional agents. The average values for four independent experiments are

Low uptake of silica nanoparticles in Caco-2 intestinal epithelial barriers

• Dong Ye,
• Mattia Bramini,
• Delyan R. Hristov,
• Sha Wan,
• Anna Salvati,
• Christoffer Åberg and
• Kenneth A. Dawson

Beilstein J. Nanotechnol. 2017, 8, 1396–1406, doi:10.3762/bjnano.8.141

• combination of imaging, flow cytometry and transport studies. Compared to typical observations in standard cell lines commonly used for in vitro studies, silica nanoparticle uptake into well-developed Caco-2 cellular barriers was found to be very low. Instead, nanoparticle association to the apical outer

• is not in direct contact with blood serum, since the cells are normally cultured in vitro in the presence of foetal bovine serum, we used foetal bovine serum as a starting point of our work. Uptake into and transport across the cellular barriers were investigated with a combination of flow cytometry

• overall barrier integrity. Nanoparticle uptake in Caco-2 barriers cultured for 4 and 21 days Flow cytometry was used to quantify cell fluorescence intensity due to association of the fluorescent SiO2-NPs with the cells. In order to test if cell polarisation/differentiation plays a role in controlling

Carbon nanomaterials sensitize prostate cancer cells to docetaxel and mitomycin C via induction of apoptosis and inhibition of proliferation

• Kati Erdmann,
• Jessica Ringel,
• Silke Hampel,
• Manfred P. Wirth and
• Susanne Fuessel

Beilstein J. Nanotechnol. 2017, 8, 1307–1317, doi:10.3762/bjnano.8.132

• Detection Kit I (BD Biosciences, Heidelberg, Germany) was used. The cells were prepared according to the recommendation of the provider followed by the analysis of 20,000 cells by flow cytometry (FACScan, BD Biosciences). Using the quadrant analysis mode of WinMDI 2.8 software (The Scripps Research

• ) served as control treatment. ap < 0.05 treatment versus control; bp < 0.05 treatment versus carbon nanomaterial alone, cp < 0.05 treatment versus chemotherapeutic alone. Exemplary flow cytometry analysis for cell death of DU-145 cells following treatment with carbon nanomaterials and chemotherapeutics

• program for women. Furthermore, the authors are grateful to Dr. Matthias Kotzsch for his technical assistance with the flow cytometry analyses.

Evaluation of quantum dot conjugated antibodies for immunofluorescent labelling of cellular targets

• Jennifer E. Francis,
• David Mason and
• Raphaël Lévy

Beilstein J. Nanotechnol. 2017, 8, 1238–1249, doi:10.3762/bjnano.8.125

• , permeabilised cells that were blocked with BSA (negatively charged). One way to assess the non-specific binding of these Qdot-Abs to cells would be to use flow cytometry. Beyond those commercially available Qdots, there have been a number of ICC protocols published for the labelling of different proteins with

Nano-engineered skin mesenchymal stem cells: potential vehicles for tumour-targeted quantum-dot delivery

• Liga Saulite,
• Dominyka Dapkute,
• Karlis Pleiko,
• Ineta Popena,
• Simona Steponkiene,
• Ricardas Rotomskis and
• Una Riekstina

Beilstein J. Nanotechnol. 2017, 8, 1218–1230, doi:10.3762/bjnano.8.123

• quantum dots QD655 were studied in human skin MSCs. The effect of QD on MSCs was examined using a cell viability assay, Ki67 expression analysis, and tri-lineage differentiation assay. The optimal conditions for QD uptake in MSCs were determined using flow cytometry. The QD uptake route in MSCs was

• markers was performed by flow cytometry. The cells were stained with CD34-PE and CD45-FITC (all from BD Biosciences, USA), CD90-FITC (Dako, USA), CD73 PE (Abcam, USA) and isotype controls IgG1-FITC (Dako, USA), IgG1-PE (BD Biosciences, USA), and IgG2A-APC (BD Biosciences, USA). Flow cytometry data were

• potential of −35.1 mV [60]. The stock solution is 8 µM in 50 mM borate, pH 9.0. Further preparations of the QD solution are described in each methodological part separately. QD uptake dynamics using flow cytometry To estimate the optimal QD concentration for uptake experiments, MSCs were seeded at a density

Surface-enhanced Raman spectroscopy of cell lysates mixed with silver nanoparticles for tumor classification

• Mohamed Hassoun,
• Iwan W.Schie,
• Tatiana Tolstik,
• Sarmiza E. Stanca,
• Christoph Krafft and
• Juergen Popp

Beilstein J. Nanotechnol. 2017, 8, 1183–1190, doi:10.3762/bjnano.8.120

• cells with subsequent microscopic assessment by pathologists. Complementary tools are immunocytochemistry, which uses fluorescence-labeled antibodies against cellular antigens, and flow cytometry, which combines several detection channels based on light scattering, absorption and fluorescence with

Uptake of the proteins HTRA1 and HTRA2 by cells mediated by calcium phosphate nanoparticles

• Olga Rotan,
• Katharina N. Severin,
• Simon Pöpsel,
• Alexander Peetsch,
• Melisa Merdanovic,
• Michael Ehrmann and
• Matthias Epple

Beilstein J. Nanotechnol. 2017, 8, 381–393, doi:10.3762/bjnano.8.40

• flow cytometry. All proteins were readily transported into the cells by cationic calcium phosphate nanoparticles. Notably, only HTRA1 was able to penetrate the cell membrane of MG-63 cells in dissolved form. However, the application of endocytosis inhibitors revealed that the uptake pathway was

• with protein or the protein alone were added to the corresponding cell samples and incubated for another 3 h. The cells were then harvested for flow cytometry analysis as follows. All samples were washed three times with PBS, detached with trypsin/EDTA (3 min at 37 °C) and transferred into 15 mL Falcon

• °C. The prepared cell samples were then transferred to flow cytometry tubes for measurements. Flow cytometry analysis Flow cytometry analysis was performed using a BD FACSCalibur™ instrument (BD Biosciences) and analysis was done using the FlowJo analysis software. Side-scatter height versus side

On the pathway of cellular uptake: new insight into the interaction between the cell membrane and very small nanoparticles

• Claudia Messerschmidt,
• Daniel Hofmann,
• Anja Kroeger,
• Katharina Landfester,
• Volker Mailänder and
• Ingo Lieberwirth

Beilstein J. Nanotechnol. 2016, 7, 1296–1311, doi:10.3762/bjnano.7.121

• to have a detailed look at the very moment of particle uptake, revealing a remarkable dependency of the observed morphologies on particle size. Alongside the EM analysis we applied confocal laser scanning microscopy (CLSM), biochemical assays for LDH and ATP, flow cytometry, Caspase-3 western

Improved biocompatibility and efficient labeling of neural stem cells with poly(L-lysine)-coated maghemite nanoparticles

• Igor M. Pongrac,
• Marina Dobrivojević,
• Lada Brkić Ahmed,
• Michal Babič,
• Miroslav Šlouf,
• Daniel Horák and
• Srećko Gajović

Beilstein J. Nanotechnol. 2016, 7, 926–936, doi:10.3762/bjnano.7.84

• was used. The quantification of cell labeling by cytometry was considered superior to Prussian blue image quantification due to the possible adherence of stain both to the cells and to the coated dish surface. By using flow cytometry and measuring the increase of the side scattered light (SSC) of the

• ®-D-spio nanoparticles, NSCs were treated with different endocytotic inhibitors, incubated with nanoparticles and subsequently, flow cytometry analysis of the labeled cells was performed (Figure 7A,B). The inhibitors were cytochalasine D (blocks actin-dependent process such as macropinocytosis

• ), nocodazole (inhibits microtubule function involved in intracellular vesicle trafficking), phenylarsine oxide (inhibits the clathrin-mediated endocytotic pathway) and filipin (inhibits caveolae pathways). Flow cytometry analysis showed that NSCs treated with cytochalasine D and incubated with PLL-γ-Fe2O3 or

PLGA nanoparticles as a platform for vitamin D-based cancer therapy

• Maria J. Ramalho,
• Joana A. Loureiro,
• Bárbara Gomes,
• Manuela F. Frasco,
• Manuel A. N. Coelho and
• M. Carmo Pereira

Beilstein J. Nanotechnol. 2015, 6, 1306–1318, doi:10.3762/bjnano.6.135

• form of calcitriol. This was the cell line where the encapsulation of calcitriol in NPs proved to be more advantageous. Cell cycle arrest by calcitriol-loaded PLGA NPs To assess whether the cytotoxic effects of calcitriol are due to cell cycle inhibition, cell cycle analysis by flow cytometry was

• the increased inhibition of cell growth in the 72 h assay, as compared with the 48 h assay. We conclude that a longstanding treatment presents more pronounced, deleterious effects since these NPs are able to maintain drug concentrations. Furthermore, flow cytometry analysis demonstrated that

• cycle analysis was conducted by flow cytometry (FCM). The cells were seeded in T75 flasks at a density of 1 × 105 cells/mL for 24 h prior to the experiment. The cells were then treated with 1.2 µM of free calcitriol and entrapped in PLGA NPs for 72 h. Due to the short half-life of calcitriol in cell

Silica micro/nanospheres for theranostics: from bimodal MRI and fluorescent imaging probes to cancer therapy

• Shanka Walia and
• Amitabha Acharya

Beilstein J. Nanotechnol. 2015, 6, 546–558, doi:10.3762/bjnano.6.57

• the synthesized nanocomposites exhibited a signal enhancement in the T1-weighted MRI images with increasing Mn concentration. The in vitro studies performed on HeLa cells suggested cell viability of more than 80% even at a Mn concentration of 50 mg·mL−1. The combination of results obtained from flow

• cytometry, confocal microscopy and MRI studies suggested that the prepared nanocomposites can be used for targeting cancer cells that overexpress folic acid. Similar strategies were also used by Peng et al. [22] by using an iridium(III) complex as fluorescent agent. Hu et al. [23] reported the synthesis of

Hematopoietic and mesenchymal stem cells: polymeric nanoparticle uptake and lineage differentiation

• Ivonne Brüstle,
• Thomas Simmet,
• Gerd Ulrich Nienhaus,
• Katharina Landfester and
• Volker Mailänder

Beilstein J. Nanotechnol. 2015, 6, 383–395, doi:10.3762/bjnano.6.38

• differentiation studies, cellular uptake and cytotoxicity of the particles were quantitatively determined by flow cytometry. After incubation with 300 µg/mL nanoparticles for 24 h, hMSCs showed a reasonable uptake of polystyrene nanoparticles (PS, Figure 1A). Since here only one population was detected in flow

• impressive and may not be strong enough to divert further development. All other expression values are not affected by the presence of these particles (Figure 7). Differentiation in flow cytometry for hHSCs Differentiation of hHSCs in three lineages was determined after incubation with 300 µg/mL

• reasonable amount by hHSCs. This is itself interesting as hHSCs have no phagocytotic properties, and the endocytotic processes in hHSCs are not known for a high turnover rate. Flow cytometry analysis showed that the particle uptake differs between the two materials, polystyrene and polylactide. Polylactide

Functionalized polystyrene nanoparticles as a platform for studying bio–nano interactions

• Cornelia Loos,
• Tatiana Syrovets,
• Anna Musyanovych,
• Volker Mailänder,
• Katharina Landfester,
• G. Ulrich Nienhaus and
• Thomas Simmet

Beilstein J. Nanotechnol. 2014, 5, 2403–2412, doi:10.3762/bjnano.5.250

• -COOH or PS-NH2 nanoparticles (each 100 µg/mL) and analyzed by flow cytometry or confocal microscopy (24 h). Results are given as mean ± SEM, n = 3, *p < 0.05, **p < 0.01. Nuclei are labeled with HCS NuclearMask™ (blue), nanoparticles are stained with PMI (green), lysosomes are labeled with LysoTracker

• stained with acridine orange (AO) and analyzed by flow cytometry. M1 gating was used to assess the number of AOlow cells with leaky lysosomes. (B) Analysis of cell viability. Cells were treated as in (A) and analyzed by XTT assay. Results are mean ± SEM, n = 3, *p < 0.05, **p < 0.01. Acknowledgements

Interaction of dermatologically relevant nanoparticles with skin cells and skin

• Annika Vogt,
• Fiorenza Rancan,
• Sebastian Ahlberg,
• Berouz Nazemi,
• Chun Sik Choe,
• Maxim E. Darvin,
• Sabrina Hadam,
• Ulrike Blume-Peytavi,
• Kateryna Loza,
• Jörg Diendorf,
• Matthias Epple,
• Christina Graf,
• Eckart Rühl,
• Martina C. Meinke and
• Jürgen Lademann

Beilstein J. Nanotechnol. 2014, 5, 2363–2373, doi:10.3762/bjnano.5.245

• cellular uptake, including optical microscopy, electron microscopy, X-ray microscopy on cells and tissue sections, flow cytometry of isolated skin cells as well as Raman microscopy on whole tissue blocks. In order to assess the biological relevance of such findings, cell viability and free radical

• with fixation and sectioning. The cells remain intact and can be analyzed by flow cytometry or single cell microscopy. For our studies on skin penetration of silica particles, we prepared single-cell suspensions of skin samples treated with fluorescent particles and performed flow cytometry and single

• /streptomycin, 2% glutamine and 10% fetal calf serum. The cells grown in an incubator with 5% CO2, 100% humidity at 37 °C and incubated with the different silica particles (10 μg/mL) for 2 h. Analysis was performed by using flow cytometry and confocal laser scanning microscopy. Figure 3 was modified with