Biology subjects

Dobson, J.

Publications and source records attributed to Dobson, J..

3 recordsLinked to original sources

bioRxiv2019

Effect of PEI-coated MNPs on the Regulation of Cellular Focal Adhesions and Actin Stress Fibres

The biocompatibility of surface coated/functionalised magnetic nanoparticles (MNPs) is key to their successful incorporation and application in biological systems. Polyethylene imine (PEI) -coated MNPs provide improved in vitro transfection efficiency compared to conventional chemical methods such as Lipofectamine and cationic polymers, and are also safer than viral transduction. Commercial cell toxicity assays are useful for end-point and high-throughput screening, providing fast results and an overview of cell health. However these assays only take into account cells that have undergone an extreme toxic response leading to cell death. Cell toxicity is a complex process which can be expressed in many forms, through morphological, metabolic, and epigenetic changes. A common indicator of cell stress and toxic response is increased cell adhesion and stress fibre formation. It is important to identify these changes in cells as it may affect downstream results and applications in biomedicine. This study explores the effect of the nanomagnetic transfection agent PEI-coated MNPs (MNP-PEIs) and an external magnetic field on cell behaviour, by studying particle internalization, changes in cellular morphology, and cell adhesion. We found that MNP-PEIs induced cell stress through a dose-dependent increase in cell adhesion via the overexpression of vinculin and formation of actin stress fibres. While the presence of PEI was the main contributor to increased cell stress, free PEI polyplexes induced higher toxicity compared to PEI bound to MNPs. MNPs without PEI coating however did not adversely affect cells suggesting a chemical effect instead of a mechanical one. In addition, genes identified as being associated with actin fibre regulation and cell adhesion, showed significant increases in expression due to the internalization of the MNP-PEI complex. From these results, we identify anomalous cell behaviour, morphology, and gene expression after interaction with MNP-PEIs, as well as a safe dosage to reduce acute cell toxicity.

bioengineering↗

bioRxiv2018

Remote manipulation of magnetic nanoparticles using magnetic field gradient to promote cancer cell death

The manipulation of magnetic nanoparticles (MNPs) using an external magnetic field, has been demonstrated to be useful in various biomedical applications. Some techniques have evolved utilizing this non-invasive external stimulus but the scientific community widely adopts few, and there is an excellent potential for more novel methods. The primary focus of this study is on understanding the manipulation of MNPs by a time-varying static magnetic field and how this can be used, at different frequencies and displacement, to manipulate cellular function. Here we explore, using numerical modeling, the physical mechanism which underlies this kind of manipulation, and we discuss potential improvements which would enhance such manipulation with its use in biomedical applications, i.e., increasing the MNP response by improving the field parameters. From our observations and other related studies, we infer that such manipulation depends mostly on the magnetic field gradient, the magnetic susceptibility and size of the MNPs, the magnet array oscillating frequency, the viscosity of the medium surrounding MNPs, and the distance between the magnetic field source and the MNPs. Additionally, we demonstrate cytotoxicity in neuroblastoma (SH-SY5Y) and hepatocellular carcinoma (HepG2) cells in vitro. This was induced by incubation with MNPs, followed by exposure to a magnetic field gradient, physically oscillating at various frequencies and displacement amplitudes. Even though this technique reliably produces MNP endocytosis and/or cytotoxicity, a better biophysical understanding is required to develop the mechanism used for this precision manipulation of MNPs, in vitro.

bioengineering↗

bioRxiv2018

Practical bioinstrumentation developments for AC magnetic field-mediated magnetic nanoparticle heating applications.

Heat dissipation during magnetization reversal processes in magnetic nanoparticles (MNP), upon exposure to alternating magnetic fields (AMF), has been extensively studied in relation to applications in magnetic fluid hyperthermia (MFH). This current paper demonstrates the design, fabrication, and evaluation of an efficient instrument, operating on this principle, for use as (i) a non-contact, in vitro, real-time temperature monitor; (ii) a drug release analysis system (DRAS); (iii) a high flux density module for AMF-mediated MNP studies; and (iv) an in vivo coil setup for real-time, whole body thermal imaging. The proposed DRAS is demonstrated by an AMF-mediated drug release proof-of-principle experiment. Also, the technique described facilitates non-contact temperature measurements of specific absorption rate (SAR) as accurately as temperature measurements using a probe in contact with the sample. Numerical calculations estimating the absolute and root mean squared flux densities, and other MNP - AMF studies suggest that the proposed stacked planar coil module could be employed for calorimetry. Even though the proposed in vivo coil setup could be used for real-time, whole body thermal imaging (within the limitations due to issues of penetration depth), further design effort is required in order to enhance the energy transfer efficiency.

bioengineering↗