Nanoparticles have become popular in life sciences in the last few years. They have been produced in many variants and have recently been used in both biological experiments and in clinical applications. Due to concerns over nanomaterial risks, there has been a dramatic increase in investigations focused on safety research. The aim of this paper is to present the advanced testing of rhodamine-derived superparamagnetic maghemite nanoparticles (SAMN-R), which are used for their nontoxicity, biocompatibility, biodegradability, and magnetic properties. Recent results were expanded upon from the basic cytotoxic tests to evaluate cell proliferation and migration potential. Two cell types were used for the cell proliferation and tracking study: mouse embryonic fibroblast cells (3T3) and human mesenchymal stem cells (hMSCs). Advanced microscopic methods allowed for the precise quantification of the function of both cell types. This study has demonstrated that a dose of nanoparticles lower than 20 µg·cm-2 per area of the dish does not negatively affect the cells' morphology, migration, cytoskeletal function, proliferation, potential for wound healing, and single-cell migration in comparison to standard CellTracker™ Green CMFDA (5-chloromethylfluorescein diacetate). A higher dose of nanoparticles could be a potential risk for cytoskeletal folding and detachment of the cells from the solid extracellular matrix.

• Klíčová slova
• cytotoxicity, fibroblast cells, magnetic nanoparticles, mesenchymal stem cells, single-cell migration, wound healing assay,
• MeSH
• biologické markery MeSH
• buněčné linie MeSH
• fibroblasty účinky léků   metabolismus   MeSH
• imunofenotypizace MeSH
• lidé MeSH
• magnetické nanočástice * chemie   MeSH
• mezenchymální kmenové buňky účinky léků   metabolismus   MeSH
• myši MeSH
• pohyb buněk účinky léků   MeSH
• proliferace buněk účinky léků   MeSH
• průtoková cytometrie MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• rhodaminy chemie   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• magnetické nanočástice * MeSH
• reaktivní formy kyslíku MeSH
• rhodaminy MeSH

Nano-ecotoxicology is extensively debated and nanomaterial surface reactivity is an emerging topic. Iron oxide nanoparticles are widely applied, with organic or inorganic coatings for stabilizing their suspensions. Surface active maghemite nanoparticles (SAMNs) are the unique example of naked iron oxide displaying high colloidal and structural stability in water and chemical reactivity. The colloidal behavior of SAMNs was studied as a function of the medium salinity and protocols of acute and chronic toxicity on Daphnia magna were consequently adapted. SAMN distribution into the crustacean, intake/depletion rates and swimming performances were evaluated. No sign of toxicity was detected in two model organisms from the first trophic level (P. subcapitata and L. minor). In D. magna, acute EC50 values of SAMN was assessed, while no sub-lethal effects were observed and the accumulation of SAMNs in the gut appeared as the sole cause of mortality. Fast depuration and absence of delayed effects indicated no retention of SAMNs within the organism. In spite of negligible toxicity on D. magna adults, SAMN surface reactivity was responsible of membrane bursting and lethality on embryos. The present study offers a contribution to the nascent knowledge concerning the impact of nanoparticle surface reactivity on biological interfaces.

• MeSH
• analýza přežití MeSH
• biotest MeSH
• chemické látky znečišťující vodu toxicita   MeSH
• Daphnia účinky léků   fyziologie   MeSH
• embryo nesavčí účinky léků   fyziologie   MeSH
• kovové nanočástice toxicita   MeSH
• lokomoce účinky léků   MeSH
• železité sloučeniny toxicita   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH
• ferric oxide MeSH Prohlížeč
• železité sloučeniny MeSH

In the last few years, magnetically labeled cells have been intensively explored, and non-invasive cell tracking and magnetic manipulation methods have been tested in preclinical studies focused on cell transplantation. For clinical applications, it is desirable to know the intracellular pathway of nanoparticles, which can predict their biocompatibility with cells and the long-term imaging properties of labeled cells. Here, we quantified labeling efficiency, localization, and fluorescence properties of Rhodamine derivatized superparamagnetic maghemite nanoparticles (SAMN-R) in mesenchymal stromal cells (MSC). We investigated the stability of SAMN-R in the intracellular space during a long culture (20 days). Analyses were based on advanced confocal microscopy accompanied by atomic absorption spectroscopy (AAS) and magnetic resonance imaging. SAMN-R displayed excellent cellular uptake (24 h of labeling), and no toxicity of SAMN-R labeling was found. 83% of SAMN-R nanoparticles were localized in lysosomes, only 4.8% were found in mitochondria, and no particles were localized in the nucleus. On the basis of the MSC fluorescence measurement every 6 days, we also quantified the continual decrease of SAMN-R fluorescence in the average single MSC during 18 days. An additional set of analyses showed that the intracellular SAMN-R signal decrease was minimally caused by fluorophore degradation or nanoparticles extraction from the cells, main reason is a cell division. The fluorescence of SAMN-R nanoparticles within the cells was detectable minimally for 20 days. These observations indicate that SAMN-R nanoparticles have a potential for application in transplantation medicine.

• Klíčová slova
• Confocal microscopy, Dual contrast agents, Intracellular fluorescent labels, Iron oxide nanoparticles, Mesenchymal stromal cells, Rhodamine, Stem cell tracking,
• MeSH
• dextrany metabolismus   MeSH
• fluorescenční spektrometrie MeSH
• intracelulární prostor metabolismus   MeSH
• lidé MeSH
• magnetické nanočástice chemie   MeSH
• mezenchymální kmenové buňky cytologie   metabolismus   MeSH
• molekulární sondy chemie   metabolismus   MeSH
• molekulární zobrazování metody   MeSH
• rhodaminy chemie   MeSH
• tuková tkáň cytologie   MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• dextrany MeSH
• ferumoxides MeSH Prohlížeč
• magnetické nanočástice MeSH
• molekulární sondy MeSH
• rhodaminy MeSH