Magnetofection is superior to other chemical transfection methods in a microglial cell line

Smolders, Silke; Kessels, Sofie; Smolders, Sophie; Poulhès, Florent; Zelphati, Olivier; Sapet, Cédric; Brône, Bert

doi:10.1016/j.jneumeth.2017.09.017

Cited by 17 publications

(8 citation statements)

References 15 publications

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“…Furthermore, Ba/F3 cells used in this study stably express TLR4–MD-2 complex differently tagged (i.e., GFP and Flag), which allowed us to easily study LPS interaction with TLR4 and receptor dimerization, avoiding transfection of primary microglia. In fact, traditional methods of microglia transfection, such as chemical-based methods, viral transduction, and electroporation, exhibit low transfection efficiency, affect cell survival, and promote inflammatory responses [55]. Furthermore, the induction of TLR4 dimerization by LPS was already shown in this cell line [18, 27].…”

Section: Discussionmentioning

confidence: 99%

Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway

Zusso

Lunardi

Franceschini

et al. 2019

J Neuroinflammation

339

209

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Background Neuroinflammation is the response of the central nervous system to events that interfere with tissue homeostasis and represents a common denominator in virtually all neurological diseases. Activation of microglia, the principal immune effector cells of the brain, contributes to neuronal injury by release of neurotoxic products. Toll-like receptor 4 (TLR4), expressed on the surface of microglia, plays an important role in mediating lipopolysaccharide (LPS)-induced microglia activation and inflammatory responses. We have previously shown that curcumin and some of its analogues harboring an α,β-unsaturated 1,3-diketone moiety, able to coordinate the magnesium ion, can interfere with LPS-mediated TLR4–myeloid differentiation protein-2 (MD-2) signaling. Fluoroquinolone (FQ) antibiotics are compounds that contain a keto-carbonyl group that binds divalent ions, including magnesium. In addition to their antimicrobial activity, FQs are endowed with immunomodulatory properties, but the mechanism underlying their anti-inflammatory activity remains to be defined. The aim of the current study was to elucidate the molecular mechanism of these compounds in the TLR4/NF-κB inflammatory signaling pathway. Methods The putative binding mode of five FQs [ciprofloxacin (CPFX), levofloxacin (LVFX), moxifloxacin, ofloxacin, and delafloxacin] to TLR4–MD-2 was determined using molecular docking simulations. The effect of CPFX and LVFX on LPS-induced release of IL-1β and TNF-α and NF-κB activation was investigated in primary microglia by ELISA and fluorescence staining. The interaction of CPFX and LVFX with TLR4–MD-2 complex was assessed by immunoprecipitation followed by Western blotting using Ba/F3 cells. Results CPFX and LVFX bound to the hydrophobic region of the MD-2 pocket and inhibited LPS-induced secretion of pro-inflammatory cytokines and activation of NF-κB in primary microglia. Furthermore, these FQs diminished the binding of LPS to TLR4–MD-2 complex and decreased the resulting TLR4–MD-2 dimerization in Ba/F3 cells. Conclusions These results provide new insight into the mechanism of the anti-inflammatory activity of CPFX and LVFX, which involves, at least in part, the activation of TLR4/NF-κB signaling pathway. Our findings might facilitate the development of new molecules directed at the TLR4–MD-2 complex, a potential key target for controlling neuroinflammation. Electronic supplementary material The online version of this article (10.1186/s12974-019-1538-9) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway

Zusso

Lunardi

Franceschini

et al. 2019

J Neuroinflammation

339

209

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“…compared the efficiency of magnetofection to other transfection methods using a microglial cell line. They found that Glial-Mag magnetofection of BV2 cells greatly outperformed standard chemical transfection methods; calcium-phosphate precipitation, X-tremeGENE, and Lipofectamine 2000 with an efficiency of 34.95% compared to 0.34, 3.30, and 12.51% respectively (Smolders et al, 2018 ). In contrast to this study, however, Katebi et al found that a static magnetic field reduces the uptake of exogenous oligonucleotide by rooster spermatozoa (Katebi et al, 2016 ).…”

Section: Magnetofectionmentioning

confidence: 99%

Force-Mediating Magnetic Nanoparticles to Engineer Neuronal Cell Function

Gahl

Kunze

2018

Front. Neurosci.

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Cellular processes like membrane deformation, cell migration, and transport of organelles are sensitive to mechanical forces. Technically, these cellular processes can be manipulated through operating forces at a spatial precision in the range of nanometers up to a few micrometers through chaperoning force-mediating nanoparticles in electrical, magnetic, or optical field gradients. But which force-mediating tool is more suitable to manipulate cell migration, and which, to manipulate cell signaling? We review here the differences in forces sensation to control and engineer cellular processes inside and outside the cell, with a special focus on neuronal cells. In addition, we discuss technical details and limitations of different force-mediating approaches and highlight recent advancements of nanomagnetics in cell organization, communication, signaling, and intracellular trafficking. Finally, we give suggestions about how force-mediating nanoparticles can be used to our advantage in next-generation neurotherapeutic devices.

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“…Magnetoporation is a well‐established magnetically guided delivery combining either chemotherapeutics or nucleic acids (e.g., anionic DNA molecules) with the cationic magnetic nanoparticles (MNPs) (commonly superparamagnetic iron oxide nanoparticles (SPIONs)) via noncovalent bonds. [ 165–167 ] By this means, first cargoes are mixed with MNPs, which are coated and tightly bonded with polyelectrolytes (i.e., polyethyleneimine). Afterward, the field‐induced transport of magnetically labeled complexes takes place through the membrane to concentrate the cargo–MNP complexes in the target cells within a few minutes.…”

Section: Macroengineered Intracellular Deliverymentioning

confidence: 99%

“…Afterward, the field‐induced transport of magnetically labeled complexes takes place through the membrane to concentrate the cargo–MNP complexes in the target cells within a few minutes. [ 167 ] Polymer nanoparticles with iron oxide cores used in this approach are preferable as they enable real‐time noninvasive monitoring of DNA delivery via magnetic resonance imaging (MRI). Accordingly, Kievit and co‐workers illustrated that SPION core coated with a novel copolymer (CP‐PEI) transfection agent is a suitable contrast enhancing agent for MRI to provide earlier detection of lesions and enhanced spatial resolution.…”

Section: Macroengineered Intracellular Deliverymentioning

confidence: 99%

A Comprehensive Review on Intracellular Delivery

Rad

Bazaz

et al. 2021

Advanced Materials

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Intracellular delivery is considered an indispensable process for various studies, ranging from medical applications (cell‐based therapy) to fundamental (genome‐editing) and industrial (biomanufacture) approaches. Conventional macroscale delivery systems critically suffer from such issues as low cell viability, cytotoxicity, and inconsistent material delivery, which have opened up an interest in the development of more efficient intracellular delivery systems. In line with the advances in microfluidics and nanotechnology, intracellular delivery based on micro‐ and nanoengineered platforms has progressed rapidly and held great promises owing to their unique features. These approaches have been advanced to introduce a smorgasbord of diverse cargoes into various cell types with the maximum efficiency and the highest precision. This review differentiates macro‐, micro‐, and nanoengineered approaches for intracellular delivery. The macroengineered delivery platforms are first summarized and then each method is categorized based on whether it employs a carrier‐ or membrane‐disruption‐mediated mechanism to load cargoes inside the cells. Second, particular emphasis is placed on the micro‐ and nanoengineered advances in the delivery of biomolecules inside the cells. Furthermore, the applications and challenges of the established and emerging delivery approaches are summarized. The topic is concluded by evaluating the future perspective of intracellular delivery toward the micro‐ and nanoengineered approaches.

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Magnetofection is superior to other chemical transfection methods in a microglial cell line

Abstract: Transfection of BV2 cells using Glial-Mag magnetofection is superior compared to other chemical transfection methods and could be considered as the method of choice to chemically transfect microglial cell lines.

Cited by 17 publications

References 15 publications

Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway

Ciprofloxacin and levofloxacin attenuate microglia inflammatory response via TLR4/NF-kB pathway

Force-Mediating Magnetic Nanoparticles to Engineer Neuronal Cell Function

A Comprehensive Review on Intracellular Delivery

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