Publications by authors named "Nathalie Luciani"

36 Publications

Massive Intracellular Remodeling of CuS Nanomaterials Produces Nontoxic Bioengineered Structures with Preserved Photothermal Potential.

ACS Nano 2021 06 25;15(6):9782-9795. Epub 2021 May 25.

Laboratoire Matière et Systèmes Complexes MSC, UMR 7057, CNRS and University of Paris, 75205, Paris Cedex 13, France.

Despite efforts in producing nanoparticles with tightly controlled designs and specific physicochemical properties, these can undergo massive nano-bio interactions and bioprocessing upon internalization into cells. These transformations can generate adverse biological outcomes and premature loss of functional efficacy. Hence, understanding the intracellular fate of nanoparticles is a necessary prerequisite for their introduction in medicine. Among nanomaterials devoted to theranostics is copper sulfide (CuS), which provides outstanding optical properties along with easy synthesis and low cost. Herein, we performed a long-term multiscale study on the bioprocessing of hollow CuS nanoparticles (CuS NPs) and rattle-like iron oxide [email protected] core-shell hybrids ([email protected] NPs) when inside stem cells and cancer cells, cultured as spheroids. In the spheroids, both CuS NPs and [email protected] NPs are rapidly dismantled into smaller units (day 0 to 3), and hair-like nanostructures are generated (day 9 to 21). This bioprocessing triggers an adaptation of the cellular metabolism to the internalized metals without impacting cell viability, differentiation, or oxidative stress response. Throughout the remodeling, a loss of IONF-derived magnetism is observed, but, surprisingly, the CuS photothermal potential is preserved, as demonstrated by a full characterization of the photothermal conversion across the bioprocessing process. The maintained photothermal efficiency correlated well with synchrotron X-ray absorption spectroscopy measurements, evidencing a similar chemical phase for Cu but not for Fe over time. These findings evidence that the intracellular bioprocessing of CuS nanoparticles can reshape them into bioengineered nanostructures without reducing the photothermal function and therapeutic potential.
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http://dx.doi.org/10.1021/acsnano.1c00567DOI Listing
June 2021

Gold-based therapy: From past to present.

Proc Natl Acad Sci U S A 2020 09 8;117(37):22639-22648. Epub 2020 Sep 8.

Laboratoire Matière et Systèmes Complexes, CNRS, Université de Paris, Paris 75205 Cedex 13, France;

Despite an abundant literature on gold nanoparticles use for biomedicine, only a few of the gold-based nanodevices are currently tested in clinical trials, and none of them are approved by health agencies. Conversely, ionic gold has been used for decades to treat human rheumatoid arthritis and benefits from 70-y hindsight on medical use. With a view to open up new perspectives in gold nanoparticles research and medical use, we revisit here the literature on therapeutic gold salts. We first summarize the literature on gold salt pharmacokinetics, therapeutic effects, adverse reactions, and the present repurposing of these ancient drugs. Owing to these readings, we evidence the existence of a common metabolism of gold nanoparticles and gold ions and propose to use gold salts as a "shortcut" to assess the long-term effects of gold nanoparticles, such as their fate and toxicity, which remain challenging questions nowadays. Moreover, one of gold salts side effects (i.e., a blue discoloration of the skin exposed to light) leads us to propose a strategy to biosynthesize large gold nanoparticles from gold salts using light irradiation. These hypotheses, which will be further investigated in the near future, open up new avenues in the field of ionic gold and gold nanoparticles-based therapies.
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http://dx.doi.org/10.1073/pnas.2007285117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7502769PMC
September 2020

Unexpected intracellular biodegradation and recrystallization of gold nanoparticles.

Proc Natl Acad Sci U S A 2020 01 18;117(1):103-113. Epub 2019 Dec 18.

Laboratoire Matière et Systèmes Complexes, CNRS, Université de Paris, Paris 75205 Cedex 13, France;

Gold nanoparticles are used in an expanding spectrum of biomedical applications. However, little is known about their long-term fate in the organism as it is generally admitted that the inertness of gold nanoparticles prevents their biodegradation. In this work, the biotransformations of gold nanoparticles captured by primary fibroblasts were monitored during up to 6 mo. The combination of electron microscopy imaging and transcriptomics study reveals an unexpected 2-step process of biotransformation. First, there is the degradation of gold nanoparticles, with faster disappearance of the smallest size. This degradation is mediated by NADPH oxidase that produces highly oxidizing reactive oxygen species in the lysosome combined with a cell-protective expression of the nuclear factor, erythroid 2. Second, a gold recrystallization process generates biomineralized nanostructures consisting of 2.5-nm crystalline particles self-assembled into nanoleaves. Metallothioneins are strongly suspected to participate in buildings blocks biomineralization that self-assembles in a process that could be affected by a chelating agent. These degradation products are similar to aurosomes structures revealed 50 y ago in vivo after gold salt therapy. Overall, we bring to light steps in the lifecycle of gold nanoparticles in which cellular pathways are partially shared with ionic gold, revealing a common gold metabolism.
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http://dx.doi.org/10.1073/pnas.1911734116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6955300PMC
January 2020

[Human stem cells can neo-biosynthesize magnetic nanoparticles after degrading man-made nanoparticles].

Med Sci (Paris) 2019 Oct 18;35(10):725-727. Epub 2019 Oct 18.

Laboratoire matière et systèmes complexes, CNRS UMR 7057, Université Paris Diderot, 10 rue Alice Domon et Léonie Duquet, 75205 Paris Cedex 13, France.

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http://dx.doi.org/10.1051/medsci/2019157DOI Listing
October 2019

Biosynthesis of magnetic nanoparticles from nano-degradation products revealed in human stem cells.

Proc Natl Acad Sci U S A 2019 03 13;116(10):4044-4053. Epub 2019 Feb 13.

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS and University Paris Diderot, 75205 Paris Cedex 13, France;

While magnetic nanoparticles offer exciting possibilities for stem cell imaging or tissue bioengineering, their long-term intracellular fate remains to be fully documented. Besides, it appears that magnetic nanoparticles can occur naturally in human cells, but their origin and potentially endogenous synthesis still need further understanding. In an effort to explore the life cycle of magnetic nanoparticles, we investigated their transformations upon internalization in mesenchymal stem cells and as a function of the cells' differentiation status (undifferentiated, or undergoing adipogenesis, osteogenesis, and chondrogenesis). Using magnetism as a fingerprint of the transformation process, we evidenced an important degradation of the nanoparticles during chondrogenesis. For the other pathways, stem cells were remarkably "remagnetized" after degradation of nanoparticles. This remagnetization phenomenon is the direct demonstration of a possible neosynthesis of magnetic nanoparticles and could lay some foundation to understand the presence of magnetic crystals in human cells. The neosynthesis was shown to take place within the endosomes and to involve the H-subunit of ferritin. Moreover, it appeared to be the key process to avoid long-term cytotoxicity (impact on differentiation) related to high doses of magnetic nanoparticles within stem cells.
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http://dx.doi.org/10.1073/pnas.1816792116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6410821PMC
March 2019

Role of growth factors and oxygen to limit hypertrophy and impact of high magnetic nanoparticles dose during stem cell chondrogenesis.

Comput Struct Biotechnol J 2018 30;16:532-542. Epub 2018 Oct 30.

Laboratoire Matière et Systèmes Complexes (MSC), UMR 7057 CNRS, University Paris Diderot, 75205 Paris cedex 13, France.

Due to an unmet clinical need of curative treatments for osteoarthritic patients, tissue engineering strategies that propose the development of cartilage tissue replacements from stem cells have emerged. Some of these strategies are based on the internalization of magnetic nanoparticles into stem cells to then initiate the chondrogenesis via magnetic compaction. A major difficulty is to drive the chondrogenic differentiation of the cells such as they produce an extracellular matrix free of hypertrophic collagen. An additional difficulty has to be overcome when nanoparticles are used, knowing that a high dose of nanoparticles can limit the chondrogenesis. We here propose a gene-based analysis of the effects of chemical factors (growth factors, hypoxia) on the chondrogenic differentiation of human mesenchymal stem cells both with and without nanoparticles. We focus on the synthesis of two of the most important constituents present in the cartilaginous extracellular matrix (Collagen II and Aggrecan) and on the expression of collagen X, the signature of hypertrophic cartilage, in order to provide a quantitative index of the type of cartilage produced (i.e. hyaline, hypertrophic). We demonstrate that by applying specific environmental conditions, gene expression can be directed toward the production of hyaline cartilage, with limited hypertrophy. Besides, a combination of the growth factors IGF-1, TGF-β3, with a hypoxic conditioning remarkably reduced the impact of high nanoparticles concentration.
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http://dx.doi.org/10.1016/j.csbj.2018.10.014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6260287PMC
October 2018

A 3D magnetic tissue stretcher for remote mechanical control of embryonic stem cell differentiation.

Nat Commun 2017 09 12;8(1):400. Epub 2017 Sep 12.

Laboratoire Matière et Systèmes Complexes (MSC), UMR 7057, CNRS and Université Paris Diderot, 75205, Paris Cedex 13, France.

The ability to create a 3D tissue structure from individual cells and then to stimulate it at will is a major goal for both the biophysics and regenerative medicine communities. Here we show an integrated set of magnetic techniques that meet this challenge using embryonic stem cells (ESCs). We assessed the impact of magnetic nanoparticles internalization on ESCs viability, proliferation, pluripotency and differentiation profiles. We developed magnetic attractors capable of aggregating the cells remotely into a 3D embryoid body. This magnetic approach to embryoid body formation has no discernible impact on ESC differentiation pathways, as compared to the hanging drop method. It is also the base of the final magnetic device, composed of opposing magnetic attractors in order to form embryoid bodies in situ, then stretch them, and mechanically stimulate them at will. These stretched and cyclic purely mechanical stimulations were sufficient to drive ESCs differentiation towards the mesodermal cardiac pathway.The development of embryoid bodies that are responsive to external stimuli is of great interest in tissue engineering. Here, the authors culture embryonic stem cells with magnetic nanoparticles and show that the presence of magnetic fields could affect their aggregation and differentiation.
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http://dx.doi.org/10.1038/s41467-017-00543-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5596024PMC
September 2017

3D Magnetic Stem Cell Aggregation and Bioreactor Maturation for Cartilage Regeneration.

J Vis Exp 2017 04 27(122). Epub 2017 Apr 27.

Laboratoire Matière et Systèmes Complexes (MSC), UMR 7057 CNRS and University Paris Diderot;

Cartilage engineering remains a challenge due to the difficulties in creating an in vitro functional implant similar to the native tissue. An approach recently explored for the development of autologous replacements involves the differentiation of stem cells into chondrocytes. To initiate this chondrogenesis, a degree of compaction of the stem cells is required; hence, we demonstrated the feasibility of magnetically condensing cells, both within thick scaffolds and scaffold-free, using miniaturized magnetic field sources as cell attractors. This magnetic approach was also used to guide aggregate fusion and to build scaffold-free, organized, three-dimensional (3D) tissues several millimeters in size. In addition to having an enhanced size, the tissue formed by magnetic-driven fusion presented a significant increase in the expression of collagen II, and a similar trend was observed for aggrecan expression. As the native cartilage was subjected to forces that influenced its 3D structure, dynamic maturation was also performed. A bioreactor that provides mechanical stimuli was used to culture the magnetically seeded scaffolds over a 21-day period. Bioreactor maturation largely improved chondrogenesis into the cellularized scaffolds; the extracellular matrix obtained under these conditions was rich in collagen II and aggrecan. This work outlines the innovative potential of magnetic condensation of labeled stem cells and dynamic maturation in a bioreactor for improved chondrogenic differentiation, both scaffold-free and within polysaccharide scaffolds.
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http://dx.doi.org/10.3791/55221DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5565124PMC
April 2017

Physiological Remediation of Cobalt Ferrite Nanoparticles by Ferritin.

Sci Rep 2017 01 9;7:40075. Epub 2017 Jan 9.

Laboratoire Matière et Systèmes Complexes, UMR 7057 CNRS/Université Paris Diderot, Sorbonne Paris Cité, 10 rue Alice Domon et Léonie Duquet, 75205 Paris Cedex 13, France.

Metallic nanoparticles have been increasingly suggested as prospective therapeutic nanoplatforms, yet their long-term fate and cellular processing in the body is poorly understood. Here we examined the role of an endogenous iron storage protein - namely the ferritin - in the remediation of biodegradable cobalt ferrite magnetic nanoparticles. Structural and elemental analysis of ferritins close to exogenous nanoparticles within spleens and livers of mice injected in vivo with cobalt ferrite nanoparticles, suggests the intracellular transfer of degradation-derived cobalt and iron, entrapped within endogenous protein cages. In addition, the capacity of ferritin cages to accommodate and store the degradation products of cobalt ferrite nanoparticles was investigated in vitro in the acidic environment mimicking the physiological conditions that are present within the lysosomes. The magnetic, colloidal and structural follow-up of nanoparticles and proteins in the lysosome-like medium confirmed the efficient remediation of nanoparticle-released cobalt and iron ions by ferritins in solution. Metal transfer into ferritins could represent a quintessential process in which biomolecules and homeostasis regulate the local degradation of nanoparticles and recycle their by-products.
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http://dx.doi.org/10.1038/srep40075DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5220348PMC
January 2017

Ferritin Protein Regulates the Degradation of Iron Oxide Nanoparticles.

Small 2017 Jan 7;13(2). Epub 2016 Nov 7.

Laboratoire Matières et Systèmes Complexes, UMR 7057 CNRS/Université Paris Diderot, Sorbonne Paris Cité, 10 rue Alice Domon et Léonie Duquet, 75205, Paris Cedex 13, France.

Proteins implicated in iron homeostasis are assumed to be also involved in the cellular processing of iron oxide nanoparticles. In this work, the role of an endogenous iron storage protein-namely the ferritin-is examined in the remediation and biodegradation of magnetic iron oxide nanoparticles. Previous in vivo studies suggest the intracellular transfer of the iron ions released during the degradation of nanoparticles to endogenous protein cages within lysosomal compartments. Here, the capacity of ferritin cages to accommodate and store the degradation products of nanoparticles is investigated in vitro in the physiological acidic environment of the lysosomes. Moreover, it is questioned whether ferritin proteins can play an active role in the degradation of the nanoparticles. The magnetic, colloidal, and structural follow-up of iron oxide nanoparticles and proteins in lysosome-like medium confirms the efficient remediation of potentially harmful iron ions generated by nanoparticles within ferritins. The presence of ferritins, however, delays the degradation of particles due to a complex colloidal behavior of the mixture in acidic medium. This study exemplifies the important implications of intracellular proteins in processes of degradation and metabolization of iron oxide nanoparticles.
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http://dx.doi.org/10.1002/smll.201602030DOI Listing
January 2017

Massive release of extracellular vesicles from cancer cells after photodynamic treatment or chemotherapy.

Sci Rep 2016 10 18;6:35376. Epub 2016 Oct 18.

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS and Université Paris Diderot, 10 rue Alice Domon et Léonie Duquet, 75205 Paris cedex 13, France.

Photodynamic therapy is an emerging cancer treatment that is particularly adapted for localized malignant tumor. The phototherapeutic agent is generally injected in the bloodstream and circulates in the whole organism as a chemotherapeutic agent, but needs light triggering to induce localized therapeutic effects. We found that one of the responses of in vitro and in vivo cancer cells to photodynamic therapy was a massive production and emission of extracellular vesicles (EVs): only 1 hour after the photo-activation, thousands of vesicles per cell were emitted in the extracellular medium. A similar effect has been found after treatment with Doxorubicin (chemotherapy), but far less EVs were produced, even 24 hours after the treatment. Furthermore, we found that the released EVs could transfer extracellular membrane components, drugs and even large intracellular objects to naive target cells. In vivo, photodynamic treatment and chemotherapy increased the levels of circulating EVs several fold, confirming the vast induction of cancer cell vesiculation triggered by anti-cancer therapies.
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http://dx.doi.org/10.1038/srep35376DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5067517PMC
October 2016

Massive Intracellular Biodegradation of Iron Oxide Nanoparticles Evidenced Magnetically at Single-Endosome and Tissue Levels.

ACS Nano 2016 08 22;10(8):7627-38. Epub 2016 Jul 22.

Laboratoire Matière et Systèmes Complexes (MSC), UMR 7057, CNRS and Université Paris Diderot , 75205 Cedex 05 Paris, France.

Quantitative studies of the long-term fate of iron oxide nanoparticles inside cells, a prerequisite for regenerative medicine applications, are hampered by the lack of suitable biological tissue models and analytical methods. Here, we propose stem-cell spheroids as a tissue model to track intracellular magnetic nanoparticle transformations during long-term tissue maturation. We show that global spheroid magnetism can serve as a fingerprint of the degradation process, and we evidence a near-complete nanoparticle degradation over a month of tissue maturation, as confirmed by electron microscopy. Remarkably, the same massive degradation was measured at the endosome level by single-endosome nanomagnetophoretic tracking in cell-free endosomal extract. Interestingly, this spectacular nanoparticle breakdown barely affected iron homeostasis: only the genes coding for ferritin light chain (iron loading) and ferroportin (iron export) were up-regulated 2-fold by the degradation process. Besides, the magnetic and tissular tools developed here allow screening of the biostability of magnetic nanomaterials, as demonstrated with iron oxide nanocubes and nanodimers. Hence, stem-cell spheroids and purified endosomes are suitable models needed to monitor nanoparticle degradation in conjunction with magnetic, chemical, and biological characterizations at the cellular scale, quantitatively, in the long term, in situ, and in real time.
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http://dx.doi.org/10.1021/acsnano.6b02876DOI Listing
August 2016

Successful chondrogenesis within scaffolds, using magnetic stem cell confinement and bioreactor maturation.

Acta Biomater 2016 06 7;37:101-10. Epub 2016 Apr 7.

Laboratoire Matière et Systèmes Complexes (MSC), UMR 7057 CNRS & University Paris Diderot, Paris F-75205 Cedex 13, France.

Unlabelled: Tissue engineering strategies, such as cellularized scaffolds approaches, have been explored for cartilage replacement. The challenge, however, remains to produce a cartilaginous tissue incorporating functional chondrocytes and being large and thick enough to be compatible with the replacement of articular defects. Here, we achieved unprecedented cartilage tissue production into a porous polysaccharide scaffold by combining of efficient magnetic condensation of mesenchymal stem cells, and dynamic maturation in a bioreactor. In optimal conditions, all the hallmarks of chondrogenesis were enhanced with a 50-fold increase in collagen II expression compared to negative control, an overexpression of aggrecan and collagen XI, and a very low expression of collagen I and RUNX2. Histological staining showed a large number of cellular aggregates, as well as an increased proteoglycan synthesis by chondrocytes. Interestingly, electron microscopy showed larger chondrocytes and a more abundant extracellular matrix. In addition, the periodicity of the neosynthesized collagen fibers matched that of collagen II. These results represent a major step forward in replacement tissue for cartilage defects.

Statement Of Significance: A combination of several innovative technologies (magnetic cell seeding, polysaccharide porous scaffolds, and dynamic maturation in bioreactor) enabled unprecedented successful chondrogenesis within scaffolds.
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http://dx.doi.org/10.1016/j.actbio.2016.04.009DOI Listing
June 2016

The One Year Fate of Iron Oxide Coated Gold Nanoparticles in Mice.

ACS Nano 2015 Aug 21;9(8):7925-39. Epub 2015 Jul 21.

Laboratoire Matières et Systèmes Complexes, UMR 7057 CNRS/Université Paris Diderot , 10 rue Alice Domon et Léonie Duquet, Paris F-75205 Cedex 13, France.

Safe implementation of nanotechnology and nanomedicine requires an in-depth understanding of the life cycle of nanoparticles in the body. Here, we investigate the long-term fate of gold/iron oxide heterostructures after intravenous injection in mice. We show these heterostructures degrade in vivo and that the magnetic and optical properties change during the degradation process. These particles eventually eliminate from the body. The comparison of two different coating shells for heterostructures, amphiphilic polymer or polyethylene glycol, reveals the long lasting impact of initial surface properties on the nanocrystal degradability and on the kinetics of elimination of magnetic iron and gold from liver and spleen. Modulation of nanoparticles reactivity to the biological environment by the choice of materials and surface functionalization may provide new directions in the design of multifunctional nanomedicines with predictable fate.
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http://dx.doi.org/10.1021/acsnano.5b00042DOI Listing
August 2015

Combining magnetic nanoparticles with cell derived microvesicles for drug loading and targeting.

Nanomedicine 2015 Apr 14;11(3):645-55. Epub 2015 Jan 14.

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS and Université Paris Diderot, France. Electronic address:

Inspired by microvesicle-mediated intercellular communication, we propose a hybrid vector for magnetic drug delivery. It consists of macrophage-derived microvesicles engineered to enclose different therapeutic agents together with iron oxide nanoparticles. Here, we investigated in vitro how magnetic nanoparticles may influence the vector effectiveness in terms of drug uptake and targeting. Human macrophages were loaded with iron oxide nanoparticles and different therapeutic agents: a chemotherapeutic agent (doxorubicin), tissue-plasminogen activator (t-PA) and two photosensitizers (disulfonated tetraphenyl chlorin-TPCS2a and 5,10,15,20-tetra(m-hydroxyphenyl)chlorin-mTHPC). The hybrid cell microvesicles were magnetically responsive, readily manipulated by magnetic forces and MRI-detectable. Using photosensitizer-loaded vesicles, we showed that the uptake of microvesicles by cancer cells could be kinetically modulated and spatially controlled under magnetic field and that cancer cell death was enhanced by the magnetic targeting. From the clinical editor: In this article, the authors devised a biogenic method using macrophages to produce microvesicles containing both iron oxide and chemotherapeutic agents. They showed that the microvesicles could be manipulated by magnetic force for targeting and subsequent delivery of the drug payload against cancer cells. This smart method could provide a novel way for future fight against cancer.
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http://dx.doi.org/10.1016/j.nano.2014.11.009DOI Listing
April 2015

[Life cycle of magnetic nanoparticles in the organism].

Biol Aujourdhui 2014 8;208(2):177-90. Epub 2014 Sep 8.

Laboratoire Matière et Systèmes Complexes, CNRS - Université Paris Diderot, 75205 Paris Cedex 13, France.

The use of nanomaterials drastically increases and yet their behavior in living organisms remains poorly examined. At the same time a better comprehension of the interactions between nanoparticles and the biological environment would allow us to limit potential nanoparticle-based toxicity and fully exploit nanoparticles medical applications. In this perspective, it is high time we develop methods to detect, quantify and follow the evolution of nanoparticles in the complex biological environment, spanning all relevant scales from the nanometer up to the tissue level. In this work we follow the life cycle of magnetic nanoparticles in vivo, focusing on their transformations over time from administration to elimination. As opposed to traditional nano-toxicological approaches, we herein take the nanoparticle perspective and try to establish how biological environment might impact the particles properties and their fate (interaction with proteins, cell confinement, degradation...) from their initial state to a series of changes a nanoparticle might undergo on its journey throughout the organism.
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http://dx.doi.org/10.1051/jbio/2014021DOI Listing
June 2015

Heat-generating iron oxide nanocubes: subtle "destructurators" of the tumoral microenvironment.

ACS Nano 2014 May 21;8(5):4268-83. Epub 2014 Apr 21.

Laboratoire Matière et Systèmes Complexes, UMR 7057 CNRS/ Université Paris Diderot , 10 rue Alice Domon et Léonie Duquet, F-75205 Paris Cedex 13, France.

Several studies propose nanoparticles for tumor treatment, yet little is known about the fate of nanoparticles and intimate interactions with the heterogeneous and ever-evolving tumor environment. The latter, rich in extracellular matrix, is responsible for poor penetration of therapeutics and represents a paramount issue in cancer therapy. Hence new strategies start aiming to modulate the neoplastic stroma. From this perspective, we assessed the efficacy of 19 nm PEG-coated iron oxide nanocubes with optimized magnetic properties to mediate mild tumor magnetic hyperthermia treatment. After injection of a low dose of nanocubes (700 μg of iron) into epidermoid carcinoma xenografts in mice, we monitored the effect of heating nanocubes on tumor environment. In comparison with the long-term fate after intravenous administration, we investigated spatiotemporal patterns of nanocube distribution, evaluated the evolution of cubes magnetic properties, and examined nanoparticle clearance and degradation processes. While inside tumors nanocubes retained their magnetic properties and heating capacity throughout the treatment due to a mainly interstitial extracellular location, the particles became inefficient heaters after cell internalization and transfer to spleen and liver. Our multiscale analysis reveals that collagen-rich tumor extracellular matrix confines the majority of nanocubes. However, nanocube-mediated hyperthermia has the potential to "destructure" this matrix and improve nanoparticle and drug penetration into neoplastic tissue. This study provides insight into dynamic interactions between nanoparticles and tumor components under physical stimulation and suggests that nanoparticle-mediated hyperthermia could be used to locally modify tumor stroma and thus improve drug penetration.
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http://dx.doi.org/10.1021/nn405356rDOI Listing
May 2014

Localization and relative quantification of carbon nanotubes in cells with multispectral imaging flow cytometry.

J Vis Exp 2013 Dec 12(82):e50566. Epub 2013 Dec 12.

Laboratoire Matière et Systèmes Complexes (MSC), CNRS/Université Paris Diderot.

Carbon-based nanomaterials, like carbon nanotubes (CNTs), belong to this type of nanoparticles which are very difficult to discriminate from carbon-rich cell structures and de facto there is still no quantitative method to assess their distribution at cell and tissue levels. What we propose here is an innovative method allowing the detection and quantification of CNTs in cells using a multispectral imaging flow cytometer (ImageStream, Amnis). This newly developed device integrates both a high-throughput of cells and high resolution imaging, providing thus images for each cell directly in flow and therefore statistically relevant image analysis. Each cell image is acquired on bright-field (BF), dark-field (DF), and fluorescent channels, giving access respectively to the level and the distribution of light absorption, light scattered and fluorescence for each cell. The analysis consists then in a pixel-by-pixel comparison of each image, of the 7,000-10,000 cells acquired for each condition of the experiment. Localization and quantification of CNTs is made possible thanks to some particular intrinsic properties of CNTs: strong light absorbance and scattering; indeed CNTs appear as strongly absorbed dark spots on BF and bright spots on DF with a precise colocalization. This methodology could have a considerable impact on studies about interactions between nanomaterials and cells given that this protocol is applicable for a large range of nanomaterials, insofar as they are capable of absorbing (and/or scattering) strongly enough the light.
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http://dx.doi.org/10.3791/50566DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4048057PMC
December 2013

High-resolution cellular MRI: gadolinium and iron oxide nanoparticles for in-depth dual-cell imaging of engineered tissue constructs.

ACS Nano 2013 Sep 20;7(9):7500-12. Epub 2013 Aug 20.

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS and Université Paris Diderot , France.

Recent advances in cell therapy and tissue engineering opened new windows for regenerative medicine, but still necessitate innovative noninvasive imaging technologies. We demonstrate that high-resolution magnetic resonance imaging (MRI) allows combining cellular-scale resolution with the ability to detect two cell types simultaneously at any tissue depth. Two contrast agents, based on iron oxide and gadolinium oxide rigid nanoplatforms, were used to "tattoo" endothelial cells and stem cells, respectively, with no impact on cell functions, including their capacity for differentiation. The labeled cells' contrast properties were optimized for simultaneous MRI detection: endothelial cells and stem cells seeded together in a polysaccharide-based scaffold material for tissue engineering appeared respectively in black and white and could be tracked, at the cellular level, both in vitro and in vivo. In addition, endothelial cells labeled with iron oxide nanoparticles could be remotely manipulated by applying a magnetic field, allowing the creation of vessel substitutes with in-depth detection of individual cellular components.
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http://dx.doi.org/10.1021/nn401095pDOI Listing
September 2013

Cell-derived vesicles as a bioplatform for the encapsulation of theranostic nanomaterials.

Nanoscale 2013 Dec 5;5(23):11374-84. Epub 2013 Jul 5.

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS and Université Paris Diderot, 10 rue Alice Domon et Léonie Duquet, 75205 Paris cedex 13, France.

There is a great deal of interest in the development of nanoplatforms gathering versatility and multifunctionality. The strategy reported herein meets these requirements and further integrates a cell-friendly shell in a bio-inspired approach. By taking advantage of a cell mechanism of biomolecule transport using vesicles, we engineered a hybrid biogenic nanoplatform able to encapsulate a set of nanoparticles regardless of their chemistry or shape. As a proof of versatility, different types of hybrid nanovesicles were produced: magnetic, magnetic-metallic and magnetic-fluorescent vesicles, either a single component or multiple components, combining the advantageous properties of each integrant nanoparticle. These nanoparticle-loaded vesicles can be manipulated, monitored by MRI and/or fluorescence imaging methods, while acting as efficient nano-heaters. The resulting assets for targeting, imaging and therapy converge for the outline of a new generation of nanosystems merging versatility and multifunctionality into a bio-camouflaged and bio-inspired approach.
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http://dx.doi.org/10.1039/c3nr01541fDOI Listing
December 2013

Magnetic and photoresponsive theranosomes: translating cell-released vesicles into smart nanovectors for cancer therapy.

ACS Nano 2013 Jun 17;7(6):4954-66. Epub 2013 May 17.

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS and Université Paris Diderot , 10 Rue Alice Domon et Léonie Duquet, 75205 Paris Cedex 13, France.

Cell-released vesicles are natural carriers that circulate in body fluids and transport biological agents to distal cells. As nature uses vesicles in cell communication to promote tumor progression, we propose to harness their unique properties and exploit these biogenic carriers as Trojan horses to deliver therapeutic payloads to cancer cells. In a theranostic approach, cell-released vesicles were engineered by a top-down procedure from precursor cells, previously loaded with a photosensitizer and magnetic nanoparticles. The double exogenous cargo provided vesicles with magnetic and optical responsiveness allowing therapeutic and imaging functions. This new class of cell-derived smart nanovectors was named "theranosomes". Theranosomes enabled efficient photodynamic tumor therapy in a murine cancer model in vivo. Moreover the distribution of this biogenic vector could be monitored by dual-mode imaging, combining fluorescence and MRI. This study reports the first success in translating a cell communication mediator into a smart theranostic nanovector.
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http://dx.doi.org/10.1021/nn400269xDOI Listing
June 2013

Real-time high-resolution magnetic resonance tracking of macrophage subpopulations in a murine inflammation model: a pilot study with a commercially available cryogenic probe.

Contrast Media Mol Imaging 2013 Mar-Apr;8(2):193-203

Laboratoire Matière et Systèmes Complexes MSC, CNRS UMR7057, Université Paris Diderot, Paris, France.

Macrophages present different polarization states exhibiting distinct functions in response to environmental stimuli. However, the dynamic of their migration to sites of inflammation is not fully elucidated. Here we propose a real-time in vivo cell tracking approach, using high-resolution (HR)-MRI obtained with a commercially available cryogenic probe (Cryoprobe™), to monitor trafficking of differently polarized macrophages after systemic injection into mice. Murine bone marrow-derived mononuclear cells were differentiated ex vivo into nonpolarized M0, pro-inflammatory M1 and immunomodulator M2 macrophage subsets and labeled with citrate-coated anionic iron oxide nanoparticles (AMNP). These cells were subsequently intravenously injected to mice bearing calf muscle inflammation. Whole body migration dynamics of macrophage subsets was monitored by MRI at 4.7 T with a volume transmission/reception radiofrequency coil and macrophage infiltration to the inflamed paw was monitored with the cryogenic probe, allowing 3D spatial resolution of 50 µm with a scan time of only 10 min. Capture of AMNP was rapid and efficient regardless of macrophage polarization, with the highest uptake in M2 macrophages. Flow cytometry confirmed that macrophages preserved their polarization hallmarks after labeling. Migration kinetics of labeled cells differed from that of free AMNP. A preferential homing of M2-polarized macrophages to inflammation sites was observed. Our in vivo HR-MRI protocol highlights the extent of macrophage infiltration to the inflammation site. Coupled to whole body imaging, HR-MRI provides quantitative information on the time course of migration of ex vivo-polarized intravenously injected macrophages.
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http://dx.doi.org/10.1002/cmmi.1516DOI Listing
July 2013

Managing magnetic nanoparticle aggregation and cellular uptake: a precondition for efficient stem-cell differentiation and MRI tracking.

Adv Healthc Mater 2013 Feb 1;2(2):313-25. Epub 2012 Nov 1.

Laboratoire Matière et Systèmes Complexes, UMR 7057 CNRS & University Paris Diderot, Paris, France.

The labeling of stem cells with iron oxide nanoparticles is increasingly used to enable MRI cell tracking and magnetic cell manipulation, stimulating the fields of tissue engineering and cell therapy. However, the impact of magnetic labeling on stem-cell differentiation is still controversial. One compromising factor for successful differentiation may arise from early interactions of nanoparticles with cells during the labeling procedure. It is hypothesized that the lack of control over nanoparticle colloidal stability in biological media may lead to undesirable nanoparticle localization, overestimation of cellular uptake, misleading MRI cell tracking, and further impairment of differentiation. Herein a method is described for labeling mesenchymal stem cells (MSC), in which the physical state of citrate-coated nanoparticles (dispersed versus aggregated) can be kinetically tuned through electrostatic and magnetic triggers, as monitored by diffusion light scattering in the extracellular medium and by optical and electronic microscopy in cells. A set of statistical cell-by-cell measurements (flow cytometry, single-cell magnetophoresis, and high-resolution MRI cellular detection) is used to independently quantify the nanoparticle cell uptake and the effects of nanoparticle aggregation. Such aggregation confounds MRI cell detection as well as global iron quantification and has adverse effects on chondrogenetic differentiation. Magnetic labeling conditions with perfectly stable nanoparticles-suitable for obtaining differentiation-capable magnetic stem cells for use in cell therapy-are subsequently identified.
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http://dx.doi.org/10.1002/adhm.201200294DOI Listing
February 2013

Intercellular carbon nanotube translocation assessed by flow cytometry imaging.

Nano Lett 2012 Sep 4;12(9):4830-7. Epub 2012 Sep 4.

CNRS/Université Paris Diderot, PRES Sorbonne-Paris Cité, Laboratoire Matière et Systèmes Complexes (MSC), UMR7057, 75205 Paris cedex 13, France.

The fate of carbon nanotubes in the organism is still controversial. Here, we propose a statistical high-throughput imaging method to localize and quantify functionalized multiwalled carbon nanotubes in cells. We give the first experimental evidence of an intercellular translocation of carbon nanotubes. This stress-induced longitudinal transfer of nanomaterials is mediated by cell-released microvesicles known as vectors for intercellular communication. This finding raises new critical issues for nanotoxicology, since carbon nanotubes could be disseminated by circulating extracellular cell-released vesicles and visiting several cells in the course of their passage into the organism.
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http://dx.doi.org/10.1021/nl302273pDOI Listing
September 2012

Adipose tissue macrophages: MR tracking to monitor obesity-associated inflammation.

Radiology 2012 Jun 20;263(3):786-93. Epub 2012 Apr 20.

INSERM, U955, Equipe 17, Créteil, 94000, France.

Purpose: To investigate whether cellular imaging by using ultrasmall superparamagnetic iron oxide (USPIO)-enhanced magnetic resonance (MR) imaging can allow detection and quantification of adipose tissue macrophage-related inflammation within adipose tissue in a mouse model.

Materials And Methods: Experimental protocols were conducted in accordance with French government policies. Adipose tissue macrophages were detected and quantified with a 4.7-T MR imager in ob/ob obese mice on the basis of the signal variance of adipose tissue triggered by injection of P904 iron oxide nanoparticles (USPIO). Mice were either intravenously injected with 1000 μmol of iron per kilogram of body weight of P904 (10 ob/ob and 11 ob/+) or used as noninjected control animals (seven ob/ob and six ob/+). Three-dimensional T2*-weighted gradient-echo MR images were acquired 10 days after intravenous injection. MR imaging signal variance in mice was correlated to adipose tissue macrophage quantification by using monoclonal antibody to F4/80 immunostaining, to proinflammatory marker quantification by using reverse transcription polymerase chain reaction (CCl2, Tnfα, Emr1), and to P904 quantification by using electron paramagnetic resonance imaging. Quantitative data were compared by using the Mann-Whitney or Student t test, and correlations were performed by using the Pearson correlation test.

Results: MR imaging measurements showed a significant increase in adipose tissue signal variance in ob/ob mice compared with ob/+ controls or noninjected animals (P < .0001), which was consistent with increased P904 uptake by adipose tissue in ob/ob mice. There was a significant and positive correlation between adipose tissue macrophage quantification at MR imaging and P904 iron oxide content (r = 0.87, P < .0001), adipose tissue macrophage-related inflammation at immunohistochemistry (r = 0.60, P < .01), and adipose tissue proinflammatory marker expression (r = 0.55, 0.56, and 0.58 for CCl2, Tnfα, and Emr1, respectively; P < .01).

Conclusion: P904 USPIO-enhanced MR imaging is potentially a tool for noninvasive assessment of adipose tissue inflammation during experimental obesity. These results provide the basis for translation of MR imaging into clinical practice as a marker of patients at risk for metabolic syndrome.
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http://dx.doi.org/10.1148/radiol.12111957DOI Listing
June 2012

Nanomagnetic sensing of blood plasma protein interactions with iron oxide nanoparticles: impact on macrophage uptake.

ACS Nano 2012 Mar 16;6(3):2665-78. Epub 2012 Feb 16.

Laboratoire Matière et Systèmes Complexes (MSC), UMR 7057 CNRS/Université Paris-Diderot, PRES Sorbonne Paris Cité, 75205 Paris cedex 13, France.

One of the first biointeractions of magnetic nanoparticles with living systems is characterized by nanoparticle-protein complex formation. The proteins dynamically encompass the particles in the protein corona. Here we propose a method based on nanomagnetism that allows a specific in situ monitoring of interactions between iron oxide nanoparticles and blood plasma. Tracking the nanoparticle orientation through their optical birefringence signal induced by an external magnetic field provides a quantitative real-time detection of protein corona at the surface of nanoparticles and assesses eventual onset of particle aggregation. Since some of the plasma proteins may cause particle aggregation, we use magnetic fractionation to separate the nanoparticle clusters (induced by "destabilizing proteins") from well-dispersed nanoparticles, which remain isolated due to a stabilizing corona involving other different types of proteins. Our study shows that the "biological identity" (obtained after the particles have interacted with proteins) and aggregation state (clustered versus isolated) of nanoparticles depend not only on their initial surface coating, but also on the concentration of plasma in the suspension. Low plasma concentrations (which are generally used in vitro) lead to different protein/nanoparticle complexes than pure plasma, which reflects the in vivo conditions. As a consequence, by mimicking in vivo conditions, we show that macrophages can perceive several different populations of nanoparticle/protein complexes (differing in physical state and in nature of associated proteins) and uptake them to a different extent. When extrapolated to what would happen in vivo, our results suggest a range of cell responses to a variety of nanoparticle/protein complexes which circulate in the body, thereby impacting their tissue distribution and their efficiency and safety for diagnostic and therapeutic use.
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http://dx.doi.org/10.1021/nn300060uDOI Listing
March 2012

Cellular transfer of magnetic nanoparticles via cell microvesicles: impact on cell tracking by magnetic resonance imaging.

Pharm Res 2012 May 21;29(5):1392-403. Epub 2012 Jan 21.

Laboratoire Matière et Systèmes Complexes, UMR 7057, CNRS/Université Paris Diderot, Sorbonne Paris Cité, 75205 Paris cedex 13, France.

Purpose: Cell labeling with magnetic nanoparticles can be used to monitor the fate of transplanted cells in vivo by magnetic resonance imaging. However, nanoparticles initially internalized in administered cells might end up in other cells of the host organism. We investigated a mechanism of intercellular cross-transfer of magnetic nanoparticles to different types of recipient cells via cell microvesicles released under cellular stress.

Methods: Three cell types (mesenchymal stem cells, endothelial cells and macrophages) were labeled with 8-nm iron oxide nanoparticles. Then cells underwent starvation stress, during which they produced microvesicles that were subsequently transferred to unlabeled recipient cells.

Results: The analysis of the magnetophoretic mobility of donor cells indicated that magnetic load was partially lost under cell stress. Microvesicles shed by stressed cells participated in the release of magnetic label. Moreover, such microvesicles were uptaken by naïve cells, resulting in cellular redistribution of nanoparticles. Iron load of recipient cells allowed their detection by MRI.

Conclusions: Cell microvesicles released under stress may be disseminated throughout the organism, where they can be uptaken by host cells. The transferred cargo may be sufficient to allow MRI detection of these secondarily labeled cells, leading to misinterpretations of the effectiveness of transplanted cells.
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http://dx.doi.org/10.1007/s11095-012-0680-1DOI Listing
May 2012

Nanomagnetism reveals the intracellular clustering of iron oxide nanoparticles in the organism.

Nanoscale 2011 Oct 20;3(10):4402-10. Epub 2011 Sep 20.

Laboratoire Matière et Systèmes Complexes, UMR 7057 CNRS/Université Paris - Diderot, 10 rue Alice Domon et Léonie Duquet, 75013 Paris, France.

There are very few methods to investigate how nanoparticles (NPs) are taken up and processed by cells in the organism in the short and long terms. We propose a nanomagnetism approach, in combination with electron microscopy, to document the magnetic outcome of iron oxide-based P904 NPs injected intravenously into mice. The NP superparamagnetic properties are shown to be modified by cell internalization, due to magnetic interactions between NPs sequestered within intracellular organelles. These modifications of magnetic behaviour are observed in vivo after NP uptake by resident macrophages in spleen and liver or by inflammatory macrophages in adipose tissue as well as in vitro in monocyte-derived macrophages. The dynamical magnetic response of cell-internalized NPs is theoretically and experimentally evidenced as a global signature of their local organization in the intracellular compartments. The clustering of NPs and their magnetism become dependent on the targeted organ, on the dose administrated and on the time elapsed since their injection. Nanomagnetism probes the intracellular clustering of iron-oxide NPs and sheds light on the impact of cellular metabolism on their magnetic responsivity.
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http://dx.doi.org/10.1039/c1nr10778jDOI Listing
October 2011

In vivo biodistribution and biological impact of injected carbon nanotubes using magnetic resonance techniques.

Int J Nanomedicine 2011 15;6:351-61. Epub 2011 Feb 15.

Université Lyon 1, Créatis-LRMN, Lyon, France.

Background: Single-walled carbon nanotubes (SWCNT) hold promise for applications as contrast agents and target delivery carriers in the field of nanomedicine. When administered in vivo, their biodistribution and pharmacological profile needs to be fully characterized. The tissue distribution of carbon nanotubes and their potential impact on metabolism depend on their shape, coating, and metallic impurities. Because standard radiolabeled or fluorescently-labeled pharmaceuticals are not well suited for long-term in vivo follow-up of carbon nanotubes, alternative methods are required.

Methods: In this study, noninvasive in vivo magnetic resonance imaging (MRI) investigations combined with high-resolution magic angle spinning (HR-MAS), Raman spectroscopy, iron assays, and histological analysis ex vivo were proposed and applied to assess the biodistribution and biological impact of intravenously injected pristine (raw and purified) and functionalized SWCNT in a 2-week longitudinal study. Iron impurities allowed raw detection of SWCNT in vivo by susceptibility-weighted MRI.

Results: A transitional accumulation in the spleen and liver was observed by MRI. Raman spectroscopy, iron assays, and histological findings confirmed the MRI readouts. Moreover, no acute toxicological effect on the liver metabolic profile was observed using the HR-MAS technique, as confirmed by quantitative real-time polymerase chain reaction analysis.

Conclusion: This study illustrates the potential of noninvasive MRI protocols for longitudinal assessment of the biodistribution of SWCNT with associated intrinsic metal impurities. The same approach can be used for any other magnetically-labeled nanoparticles.
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http://dx.doi.org/10.2147/IJN.S16653DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3075901PMC
August 2011

Long term in vivo biotransformation of iron oxide nanoparticles.

Biomaterials 2011 Jun;32(16):3988-99

Laboratoire Matières et Systèmes Complexes (MSC), UMR 7057 CNRS/Université Paris-Diderot, 10 Rue Alice Domon et Léonie Duquet, 75205 Paris Cedex 13, France.

The long term outcome of nanoparticles in the organism is one of the most important concerns raised by the development of nanotechnology and nanomedicine. Little is known on the way taken by cells to process and degrade nanoparticles over time. In this context, iron oxide superparamagnetic nanoparticles benefit from a privileged status, because they show a very good tolerance profile, allowing their clinical use for MRI diagnosis. It is generally assumed that the specialized metabolism which regulates iron in the organism can also handle iron oxide nanoparticles. However the biotransformation of iron oxide nanoparticles is still not elucidated. Here we propose a multiscale approach to study the fate of nanomagnets in the organism. Ferromagnetic resonance and SQUID magnetization measurements are used to quantify iron oxide nanoparticles and follow the evolution of their magnetic properties. A nanoscale structural analysis by electron microscopy complements the magnetic follow-up of nanoparticles injected to mice. We evidence the biotransformation of superparamagnetic maghemite nanoparticles into poorly-magnetic iron species probably stored into ferritin proteins over a period of three months. A putative mechanism is proposed for the biotransformation of iron-oxide nanoparticles.
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http://dx.doi.org/10.1016/j.biomaterials.2011.02.031DOI Listing
June 2011
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