Influence of gold, silver and gold–silver alloy nanoparticles on germ cell function and embryo development

• Ulrike Taylor,
• Daniela Tiedemann,
• Christoph Rehbock,
• Wilfried A. Kues,
• Stephan Barcikowski and
• Detlef Rath

Beilstein J. Nanotechnol. 2015, 6, 651–664, doi:10.3762/bjnano.6.66

• traditionally used in somatic cell culture. Table 1 gives examples of common reprotoxicological in vitro tests and their predictive values. However, despite its obvious importance, reprotoxicological testing of nanoparticles has so far been frequently neglected. While in the last two years approximately 1000

Novel ZnO:Ag nanocomposites induce significant oxidative stress in human fibroblast malignant melanoma (Ht144) cells

• Syeda Arooj,
• Samina Nazir,
• Akhtar Nadhman,
• Nafees Ahmad,
• Bakhtiar Muhammad,
• Ishaq Ahmad,
• Kehkashan Mazhar and
• Rashda Abbasi

Beilstein J. Nanotechnol. 2015, 6, 570–582, doi:10.3762/bjnano.6.59

• of the nanocomposites were calculated by the Kubelka–Munk function F(R) by using the following equation [30]. where F(R) is the Kubelka–Munk function; R represents the absolute value of reflectance. Cell lines and cell culture Human malignant melanoma (HT144, ATCC HTB-63) cells were cultured in RPMI

• in de-ionized water and dispersed by sonication on sonifier cell disrupter (40 W, 25 kHz) for 30 min in a water bath. The nanoparticle serial dilutions were prepared as required by using the appropriate cell culture medium. To screen the photo-oxidation-mediated effects of the NPs, briefly, 1.5 × 105

Pulmonary surfactant augments cytotoxicity of silica nanoparticles: Studies on an in vitro air–blood barrier model

• Jennifer Y. Kasper,
• Lisa Feiden,
• Maria I. Hermanns,
• Christoph Bantz,
• Michael Maskos,
• Ronald E. Unger and
• C. James Kirkpatrick

Beilstein J. Nanotechnol. 2015, 6, 517–528, doi:10.3762/bjnano.6.54

• nanoparticles. Particle size was determined using the method of dynamic light scattering (DLS). Thus, the reported sizes are z-weighted mean values of the hydrodynamic diameter. Particle diameters were measured in cell culture medium (RPMI 1640) and, for reference, in water (containing 2 mmol/L sodium bromide

• change in particle size could be detected for any of the particle types in water as well as in RPMI 1640 cell culture medium. Furthermore, no significantly different particle sizes were measured for any of the different surface modifications. As the particles exhibit comparable sizes and an identical

• nanometers in diameter were found in cell culture medium (data not shown). However, the analysis of the pure Alveofact® dispersion revealed that agglomerates of the same size were already present without nanoparticles. This leads to the conclusion that the standard procedure proposed by the manufacturer for

A surface acoustic wave-driven micropump for particle uptake investigation under physiological flow conditions in very small volumes

• Florian G. Strobl,
• Dominik Breyer,
• Phillip Link,
• Adriano A. Torrano,
• Christoph Bräuchle,
• Matthias F. Schneider and
• Achim Wixforth

Beilstein J. Nanotechnol. 2015, 6, 414–419, doi:10.3762/bjnano.6.41

• uptake of nanoparticles. Here, we present a versatile microfluidic device based on acoustic streaming induced by surface acoustic waves (SAWs). The device offers a convenient method for introducing fluid motion in standard cell culture chambers and for mimicking capillary blood flow. We show that shear

• to the aforementioned problems is to perform experiments under realistic flow conditions. In the following, we introduce a novel device for inducing high shear rates at the bottom of an arbitrary cell culture chamber. The device is based on SAW-driven acoustic streaming. Many different applications

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 was induced using NH Osteo Diff Medium/NH Chrondro Diff Medium (Miltenyi Biotec Bergisch Gladbach, Germany); the medium was changed twice a week. Analysis was performed after 24 days of cell culture. Before starting the differentiation, all cells were incubated with 300 µg/mL nanoparticles for 24

• the d0 sample as 1. The relative expression was used if d0 samples did not show expression of the used marker gene and was calculated with the ΔCT method, setting the expression in the sample without particles to 1. HTRF Assay The levels of IL-8 in the cell culture supernatant were quantitatively

• performed after 5 days of cultivation with a HTRF assay by analyzing the cell culture supernatant. p > 0.05 ns, p < 0.001 ***. Cytochemical staining to determine the differentiation of hMSCs incubated with different nanoparticles (300 µg/mL nanoparticles, incubation time 24 h, before inducing

Comparative evaluation of the impact on endothelial cells induced by different nanoparticle structures and functionalization

• Lisa Landgraf,
• Ines Müller,
• Peter Ernst,
• Miriam Schäfer,
• Christina Rosman,
• Isabel Schick,
• Oskar Köhler,
• Hartmut Oehring,
• Vladimir V. Breus,
• Thomas Basché,
• Carsten Sönnichsen,
• Wolfgang Tremel and
• Ingrid Hilger

Beilstein J. Nanotechnol. 2015, 6, 300–312, doi:10.3762/bjnano.6.28

• according to the experiment and the cell line. Cell culture The mouse endothelial (SVEC4-10) cell line was obtained from American Type Culture Collection (LGC Standards GmbH, Germany). Cells were grown in Dulbecco´s modified eagle´s medium (DMEM with Glutamax; Invitrogen, Germany) supplemented with 10% heat

The effect of surface charge on nonspecific uptake and cytotoxicity of CdSe/ZnS core/shell quantum dots

• Vladimir V. Breus,
• Anna Pietuch,
• Marco Tarantola,
• Thomas Basché and
• Andreas Janshoff

Beilstein J. Nanotechnol. 2015, 6, 281–292, doi:10.3762/bjnano.6.26

• (Figure 3a). The growth of CA–QDs aggregates over time could be caused by opsonization (e.g., the adsorption of proteins from the cell culture medium on the nanoparticles). Serum-induced aggregation was reported earlier for CdSe/ZnS QDs with cationic charge [19]. Some very large fluorescent aggregates can

• early stages. This work highlights the importance of the combined use of ECIS and MTS (or other biochemical) assays for the characterization of nanoparticle cytotoxicity. Experimental Cell culture MDCKII cells were maintained in Earle’s minimum essential medium supplemented with 4 mM glutamine, 100 g/mL

• , followed by trypsinization and centrifugation at 110g. Counting was carried out using a Neubauer chamber, and viability was determined using trypan blue exclusion. For fluorescence microscopy measurements, the cells were grown in 2 mL of cell culture medium in a ibiTreat µ-Dish (Ibidi, Martinsried, Germany

Release behaviour and toxicity evaluation of levodopa from carboxylated single-walled carbon nanotubes

• Julia M. Tan,
• Jhi Biau Foo,
• Sharida Fakurazi and
• Mohd Zobir Hussein

Beilstein J. Nanotechnol. 2015, 6, 243–253, doi:10.3762/bjnano.6.23

• Mytilineou et al. [30], where the LD exposed to mesencephalic cultures at 72 h of treatment was associated with a dose-dependent reduction in cell survival. The authors attributed the potential toxicity of LD to the decreased antioxidant capacity in the limited environment of the cell culture that makes the

Tailoring the ligand shell for the control of cellular uptake and optical properties of nanocrystals

• Johannes Ostermann,
• Christian Schmidtke,
• Christopher Wolter,
• Jan-Philip Merkl,
• Hauke Kloust and
• Horst Weller

Beilstein J. Nanotechnol. 2015, 6, 232–242, doi:10.3762/bjnano.6.22

• copolymers. Tested samples and the observed interaction with A549 cells. Supporting Information Supporting Information File 96: Additional information about the conducted cell culture experiments available. Acknowledgements This work was supported by the Deutsche Forschungsgemeinschaft (DFG, SPP 1313), the

Caveolin-1 and CDC42 mediated endocytosis of silica-coated iron oxide nanoparticles in HeLa cells

• Nils Bohmer and
• Andreas Jordan

Beilstein J. Nanotechnol. 2015, 6, 167–176, doi:10.3762/bjnano.6.16

• , the fluorescent dye Alexa Fluor® 555 was embedded into their shells. Further details of synthesis and characterization have been described elsewhere [29][30]. Cell culture HeLa cells (human cervix carcinoma) were provided by the group of Professor Haucke (Freie Universität, Berlin, Germany). They were

• 70–80% confluence. Before every single experiment, nanoparticles were prewarmed to 37 °C and treated with ultrasound for 10 min to avoid sedimentation and aggregation. SPIONs were diluted with cell culture media to a concentration of 50 µg Fe/mL while SCIONs were diluted to 5 µg/mL. The concentration

• of the SCIONs was chosen after preliminary experiments. It could be shown that 5 µg Fe/mL provides the lowest background fluorescence combined with a good intracellular signal. In both setups the cell culture media contains SCIONs in excess to the internalization rate of the cells. Afterward cells

Increasing throughput of AFM-based single cell adhesion measurements through multisubstrate surfaces

• Miao Yu,
• Nico Strohmeyer,
• Jinghe Wang,
• Daniel J. Müller and
• Jonne Helenius

Beilstein J. Nanotechnol. 2015, 6, 157–166, doi:10.3762/bjnano.6.15

• hydrophilic during plasma treatment, and thus, liquid coating drops spread over the masks. Storing the PDMS in air will restore the hydrophobicity of the surface. Cell culture PC3 cells were maintained in RPMI-1640-supplemented (Gibco-Life technologies) 1 mM sodium pyruvate; HeLa cells (Kyoto) and mouse

The distribution and degradation of radiolabeled superparamagnetic iron oxide nanoparticles and quantum dots in mice

• Denise Bargheer,
• Artur Giemsa,
• Barbara Freund,
• Markus Heine,
• Christian Waurisch,
• Gordon M. Stachowski,
• Stephen G. Hickey,
• Alexander Eychmüller,
• Jörg Heeren and
• Peter Nielsen

Beilstein J. Nanotechnol. 2015, 6, 111–123, doi:10.3762/bjnano.6.11

• various Qdots in cell culture [7][8][9][10][11][12]. However, when dosing was applied under physiological conditions in mice and rats, no abnormal behavior or tissue damage was observed over the period of months after systemic administration of Qdots [13][14][15]. Therefore, the relevancy of the in vitro

• cellular internalization and processing of polymer-coated Qdots and SPIOs was gained by cell culture. For comparison of the in vivo data, these studies were made with J774 cells (a murine macrophage cell line). The cells were incubated with polymer-coated Qdots for 2 h, then the nanoparticles were removed

• µm thicknesses were cut from frozen specimens embedded in an optimum cutting temperature formulation solution (Sakura Finetek Europe). Cryosections were mounted on a slide (Superfrost/Plus, Glaswarenfabrik Karl Hecht KG, Sondheim, Germany) and dried overnight before immunostaining. Cell culture J774

The fate of a designed protein corona on nanoparticles in vitro and in vivo

• Denise Bargheer,
• Julius Nielsen,
• Gabriella Gébel,
• Markus Heine,
• Sunhild C. Salmen,
• Roland Stauber,
• Horst Weller,
• Joerg Heeren and
• Peter Nielsen

Beilstein J. Nanotechnol. 2015, 6, 36–46, doi:10.3762/bjnano.6.5

• transferrin receptor binding on the surface of cells [23]. It should be noted that most of the mentioned work was performed in in vitro experiments using partly also cell culture models. To date, much less information is available on the consequences of protein adsorption on nanoparticles in vivo. This is not

• charged SPIOs had no preformed transferrin corona but must have been also coated by plasma proteins before liver uptake took place implicating a similar biodistribution. This confirms the stability of a corona in a recent study by Wang et al. using a cell culture model [22]. Conclusion This pilot study

Mammalian cell growth on gold nanoparticle-decorated substrates is influenced by the nanoparticle coating

• Christina Rosman,
• Sebastien Pierrat,
• Marco Tarantola,
• David Schneider,
• Eva Sunnick,
• Andreas Janshoff and
• Carsten Sönnichsen

Beilstein J. Nanotechnol. 2014, 5, 2479–2488, doi:10.3762/bjnano.5.257

• ]). Cell culture In our studies, we used epithelial MDCK (type II) cells and performed cell culture as described in [20]. Cells were cultured in Earle’s minimum essential medium supplemented with glutamine (4 mM), penicillin and streptomycin (100 µg/mL for both), fetal calf serum (10% v/v) and stored in an

• optical dark field microscopy measurements. Finally, 10 μL of the cell suspension (≈6000 cells) were seeded and samples were kept under cell culture conditions. Live cell imaging Optical dark field microscopy was performed after 4, 24, 48, and 72 h while maintaining 37 °C and a 5% CO2 environment. In

• order to improve contrast for imaging, the cell culture medium was replaced by a version without a phenol red pH indicator or any nutrients. As soon as imaging was finished, the medium was changed back to the full culture medium and the samples were kept in a cell culture incubator. Electric cell

Proinflammatory and cytotoxic response to nanoparticles in precision-cut lung slices

• Stephanie Hirn,
• Nadine Haberl,
• Kateryna Loza,
• Matthias Epple,
• Wolfgang G. Kreyling,
• Barbara Rothen-Rutishauser,
• Markus Rehberg and
• Fritz Krombach

Beilstein J. Nanotechnol. 2014, 5, 2440–2449, doi:10.3762/bjnano.5.253

• testing the toxicity of nanoparticles. In vitro test systems, offering the advantages of easy handling and high-trough-put screening, can provide a first estimation of possible toxic effects. However, cell culture systems do not adequately reflect the real in vivo situation, whereas animal experiments are

• material [38][39]. Dissolution and therefore release of Zn2+ ions in the culture medium has been already described for the ZnO-NPs used in the present study [40]. Moreover, the dissolution (50 to 60% within 24 h) of ZnO-NPs in cell culture medium facilitates an interaction of Zn2+ ions with cells [27]. Ag

• ], viability was determined as ratio of nuclei of dead cells per volume of live cells (spots (diameter ≤ 4 µm)/volume (105 µm3). WST-1 reduction in PCLS culture medium The WST-1 assay (Roche Diagnostics, Mannheim, Germany) was used for spectrometric quantification of cellular viability. After removing cell

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

• about four times more negatively charged nanoparticles in cell culture medium. By contrast, monocytic leukemia cells, internalized more rapidly positively charged nanoparticles independently of the assay media. The ability of macrophages to preferentially internalize negatively charged nanoparticles

• , positively charged particles were found mostly in leukemia xenografts [43]. With particles left in the cell culture media, the cellular uptake reaches equilibrium within 24 h. When particles were removed from the media, there was virtually an exponential decrease of the amount of particles in proliferating

• % of PS-NH2 particles left. Neither THP-1, nor macrophages release nanoparticles back into the culture media, as measured during the six days of cell culture. This indicates that in macrophages, which do not proliferate, the amount of internalized non-biodegradable nanoparticles is not reduced with

Nanoparticle interactions with live cells: Quantitative fluorescence microscopy of nanoparticle size effects

• Li Shang,
• Karin Nienhaus,
• Xiue Jiang,
• Linxiao Yang,
• Katharina Landfester,
• Volker Mailänder,
• Thomas Simmet and
• G. Ulrich Nienhaus

Beilstein J. Nanotechnol. 2014, 5, 2388–2397, doi:10.3762/bjnano.5.248

• -(2,6-diisopropylphenyl)-perylene-3,4-dicarbonacidimide (PMI, BASF, Ludwigshafen, Germany), were synthesized by a miniemulsion polymerization approach [30]. Cell culture HeLa cells were cultured in Dulbecco’s modified eagle medium (DMEM), supplemented with 10% fetal bovine serum (FBS), 60 µg/mL

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

• Temperature Keratinocytes) and primary human keratinocytes showed an increased uptake of silica particles with positive surface charge under cell culture conditions, which was due to functionalization, e.g., through (3-aminopropyl)triethoxysilane (APS) (Figure 3) [3]. In the case of these silica particles

• heavily aggregated but still taken up into cells in large numbers. However, N-(6-aminohexyl)-aminopropyltrimethoxysilane (AHAPS)-functionalized particles, which had also a highly positive zeta-potential due to the amino groups but did not aggregate in cell culture media were also found in large numbers in

• the cells [35]. In our studies on the stability of differently functionalized silica particles, even different standard cell culture media compositions resulted in different aggregation behaviors of nanoparticle preparations [35]. The multitude of possible interactions on the skin surface and in the

Coating with luminal gut-constituents alters adherence of nanoparticles to intestinal epithelial cells

• Heike Sinnecker,
• Katrin Ramaker and
• Andreas Frey

Beilstein J. Nanotechnol. 2014, 5, 2308–2315, doi:10.3762/bjnano.5.239

• encounter in the intestinal tract may have on particle adherence to epithelial cells. The study using an in vitro cell culture as model system was intended to reveal at least some initial information about this very complex situation. Results and Discussion To investigate the interaction of NPs with

• GmbH, Darmstadt, Germany). Cell culture medium and supplements were obtained from PAA Laboratories GmbH (Pasching, Austria) and Roche Applied Science (Mannheim, Germany). Native intestinal fluid was harvested from the dissected small intestine of Balb/c mice as described previously [30]. Cell culture

Anticancer efficacy of a supramolecular complex of a 2-diethylaminoethyl–dextran–MMA graft copolymer and paclitaxel used as an artificial enzyme

• Yasuhiko Onishi,
• Yuki Eshita,
• Rui-Cheng Ji,
• Masayasu Onishi,
• Takashi Kobayashi,
• Masaaki Mizuno,
• Jun Yoshida and
• Naoji Kubota

Beilstein J. Nanotechnol. 2014, 5, 2293–2307, doi:10.3762/bjnano.5.238

• expressed by enzymatic kinetic parameters. Relationship between Cd and paclitaxel concentration, [E]0, after 24 h and 48 h. After 24 h: Cd = 0.1062 [E]0 + 0.0481; after 48 h: Cd = 0.1156 [E]0 + 0.1082. (c) Apparatus constant (C1) by cell culture time in WST8 vs time (h) for DDMC/PTX complex. Reprinted from

The gut wall provides an effective barrier against nanoparticle uptake

• Heike Sinnecker,
• Thorsten Krause,
• Sabine Koelling,
• Ingmar Lautenschläger and
• Andreas Frey

Beilstein J. Nanotechnol. 2014, 5, 2092–2101, doi:10.3762/bjnano.5.218

• model the structural barrier of a healthy gut is retained without the complexity of a whole animal model or the simplifications of a mere cell culture system. Our results obtained with this ex vivo system show that the multiple fencings of the intestinal mucosa combine into an effective barrier against

Effect of silver nanoparticles on human mesenchymal stem cell differentiation

• Christina Sengstock,
• Jörg Diendorf,
• Matthias Epple,
• Thomas A. Schildhauer and
• Manfred Köller

Beilstein J. Nanotechnol. 2014, 5, 2058–2069, doi:10.3762/bjnano.5.214

• microscopy, photometry and by analyzing the secretion of typical biomarkers. Results Uptake and intracellular distribution of nano-silver in hMSCs Human MSCs were cultured in the presence of 20 µg·mL−1 Ag-NP at 37 °C for 24 h under cell culture conditions, and the cell nucleus and endo-lysosomes were labeled

• with specific organelle markers. Laser scanning microscopy and phase-contrast microscopy were performed in parallel on identical cell areas (Figure 1A). In cells cultured in the presence of Ag-NPs, agglomerated nanoparticles were visible in a region close to the cell nucleus but not in the cell culture

• dihydrate (Fluka, p.a.), silver nitrate (Fluka, p.a.), and D-(+)-glucose (Baker) were used. Ultrapure water was prepared with an ELGA Purelab ultra instrument. Ag-NP were stored under argon to prevent partial oxidative dissolution (which drastically influences nanoparticle toxicity) prior to cell culture

Rapid degradation of zinc oxide nanoparticles by phosphate ions

• Rudolf Herrmann,
• F. Javier García-García and
• Armin Reller

Beilstein J. Nanotechnol. 2014, 5, 2007–2015, doi:10.3762/bjnano.5.209

• nanoparticles are highly sensitive towards phosphate ions even at pH 7. Buffer solutions and cell culture media containing phosphate ions are able to destroy ZnO nanoparticles within a time span from less than one hour to one day. The driving force of the reaction is the formation of zinc phosphate of very low

• visible range when excited in the UV region [1]. During first tests of ZnO-NP for their interaction with biological systems in the SPP 1313 priority programme (BioNanoResponses) we observed that this fluorescence is rapidly (within seconds) quenched in some cell culture media. Tracing the effect back to

• cosmetics, studies on interactions with biological systems are necessary. Cell culture media contain varying amounts of phosphate ions, from low (less than 100 mg/L, as in human blood) to high (more than 1000 mg/L for CO2-independent media). One can therefore expect that the choice of the cell culture

PVP-coated, negatively charged silver nanoparticles: A multi-center study of their physicochemical characteristics, cell culture and in vivo experiments

• Sebastian Ahlberg,
• Alexandra Antonopulos,
• Jörg Diendorf,
• Ralf Dringen,
• Matthias Epple,
• Rebekka Flöck,
• Wolfgang Goedecke,
• Christina Graf,
• Nadine Haberl,
• Jens Helmlinger,
• Fabian Herzog,
• Frederike Heuer,
• Stephanie Hirn,
• Christian Johannes,
• Stefanie Kittler,
• Manfred Köller,
• Katrin Korn,
• Wolfgang G. Kreyling,
• Fritz Krombach,
• Jürgen Lademann,
• Kateryna Loza,
• Eva M. Luther,
• Marcelina Malissek,
• Martina C. Meinke,
• Daniel Nordmeyer,
• Anne Pailliart,
• Jörg Raabe,
• Fiorenza Rancan,
• Barbara Rothen-Rutishauser,
• Eckart Rühl,
• Carsten Schleh,
• Andreas Seibel,
• Christina Sengstock,
• Lennart Treuel,
• Annika Vogt,
• Katrin Weber and
• Reinhard Zellner

Beilstein J. Nanotechnol. 2014, 5, 1944–1965, doi:10.3762/bjnano.5.205

• silver nanoparticles in vivo induced a moderate pulmonary toxicity, however, only at rather high concentrations. The same was found in precision-cut lung slices of rats in which silver nanoparticles remained mainly at the tissue surface. In a human 3D triple-cell culture model consisting of three cell

• as reducing sugar, and cysteine as a model compound for sulfur-containing proteins. H2O2 was used as strongly oxidizing compound. Furthermore, we analyzed the behavior of silver nanoparticles after their immersion in cell culture media (DMEM, RPMI and LB medium) [20]. Figure 3A shows that the

• under cell culture conditions. This distinction was made in order to separate the effect of particles from that of ions. In the presence of silver nanoparticles, agglomerated nanoparticles were detectable in a region close to the cell nucleus (Figure 6B). In contrast, agglomerates were not observed

Imaging the intracellular degradation of biodegradable polymer nanoparticles

• Anne-Kathrin Barthel,
• Martin Dass,
• Melanie Dröge,
• Jens-Michael Cramer,
• Daniela Baumann,
• Markus Urban,
• Katharina Landfester,
• Volker Mailänder and
• Ingo Lieberwirth

Beilstein J. Nanotechnol. 2014, 5, 1905–1917, doi:10.3762/bjnano.5.201

• : decomposition of the PLLA nanoparticles and mitosis. The latter process will reduce the average fluorescence intensity of the cell culture by a factor of two for every cell division. The individual PLLA nanoparticle distribution among the two daughter cells, however, might be highly asymmetric [29]. Hence, the

• scanning microscopy (CLSM) In order to investigate the intracellular localization of the PLLA nanoparticles, CLSM investigation of the cell culture was conducted concurrently with the flow cytometry measurements. Figure 2 shows the CLSM micrographs of the MSCs at different times after starting the