Nanoparticles engineered for biomedical applications are meant to be in contact with protein-rich physiological fluids. These proteins are usually adsorbed onto the nanoparticle's surface, forming a swaddling layer that has been described as a 'protein corona', the nature of which is expected to influence not only the physicochemical properties of the particles but also the internalization into a given cell type. We have investigated the process of protein adsorption onto different magnetic nanoparticles (MNPs) when immersed in cell culture medium, and how these changes affect the cellular uptake. The role of the MNPs surface charge has been assessed by synthesizing two colloids with the same hydrodynamic size and opposite surface charge: magnetite (Fe3O4) cores of 25-30 nm were in situ functionalized with (a) positive polyethyleneimine (PEI-MNPs) and (b) negative poly(acrylic acid) (PAA-MNPs). After few minutes of incubation in cell culture medium the wrapping of the MNPs by protein adsorption resulted in a 5-fold increase of the hydrodynamic size. After 24 h of incubation large MNP-protein aggregates with hydrodynamic sizes of ≈1500 nm (PAA-MNPs) and ≈3000 nm (PEI-MNPs) were observed, each one containing an estimated number of magnetic cores between 450 and 1000. These results are consistent with the formation of large protein-MNPs aggregate units having a 'plum pudding' structure of MNPs embedded into a protein network that results in a negative surface charge, irrespective of the MNP-core charge. In spite of the similar negative ζ-potential for both MNPs within cell culture, we demonstrated that PEI-MNPs are incorporated in much larger amounts than the PAA-MNPs units. Quantitative analysis showed that SH-SY5Y cells can incorporate 100% of the added PEI-MNPs up to ≈100 pg/cell, whereas for PAA-MNPs the uptake was less than 50%. The final cellular distribution showed also notable differences regarding partial attachment to the cell membrane. These results highlight the need to characterize the final properties of MNPs after protein adsorption in biological media, and demonstrate the impact of these properties on the internalization mechanisms in neural cells.

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Carbon nanotubes (CNTs) are widely used for biomedical applications as intracellular transporters of biomolecules owing to their ability to cross cell membranes. In this article, we survey the reported literature and results of our published work in an attempt to provide a rational view of the various CNT internalization mechanisms. Essentially three uptake mechanisms (phagocytosis, diffusion and endocytosis) have been reported in the literature. In addressing the subject of cellular internalization of CNTs, the unique physicochemical characteristics of CNTs that influence and drive the cell uptake pathway are considered. According to available evidence, the degree of dispersion, the formation of supramolecular complexes and the nanotube length are crucial factors in determining the exact mechanism of cellular uptake. In conclusion, phagocytosis appears to be the internalization pathway for CNT aggregates, bundles, cluster or single dispersed nanotubes 1 microm or more in length; endocytosis is the internalization mechanism for nanotubes forming supramolecular structures; and diffusion is the internalization mechanism for submicron CNTs that do not form supramolecular complexes. This information may be relevant to the rational design of CNT-based carriers for cell therapy.

We report a one-step synthesis protocol for obtaining polymer-coated magnetic nanoparticles (MNPs) engineered for uploading neural cells. Polyethyleneimine-coated Fe3O4 nanoparticles (PEI-MNPs) with sizes of 25 ± 5 nm were prepared by oxidation of Fe(OH)2 by nitrate in basic aqueous media and adding PEI in situ during synthesis. The obtained PEI-MNP cores displayed a neat octahedral morphology and high crystallinity. The resulting nanoparticles were coated with a thin polymer layer of about 0.7-0.9 nm, and displayed a saturation magnetization value M S = 58 A m2 kg-1 at 250 K (64 A m2 kg-1 for T = 10 K). Cell uptake experiments on a neuroblastoma- derived SH-SY5Y cell line were undertaken over a wide time and MNP concentration range. The results showed a small decrease in cell viability for 24 h incubation (down to 70% viability for 100 μg ml-1), increasing the toxic effects with incubation time (30% cell survival at 100 μg ml -1 for 7 days of incubation). On the other hand, primary neuronal cells displayed higher sensitivity to PEI-MNPs, with a cell viability reduction of 44% of the control cells after 3 days of incubation with 50 μg ml -1. The amount of PEI-MNPs uploaded by SH-SY5Y cells was found to have a linear dependence on concentration. The intracellular distribution of the PEI-MNPs analyzed at the single-cell level by the dual-beam (FIB/SEM) technique revealed the coexistence of both fully incorporated PEI-MNPs and partially internalized PEI-MNP-clusters crossing the cell membrane. The resulting MNP-cluster distributions open the possibility of using these PEI-MNPs for magnetically driven axonal re-growth in neural cells

This review based on the Wickham lecture given by AC at the 2009 SMIT meeting in Sinaia outlines the progress made in nano-technology for healthcare. It describes in brief the nature of nano-materials and their unique properties which accounts for the significant research both in scientific institutions and industry for translation into new therapies embodied in the emerging field of nano-medicine. It stresses that the potential of nano-medicine to make significant inroads for more effective therapies both for life-threatening and life-disabling disorders will only be achieved by high-quality life science research. The first generation of passive nano-diagnostics based on nanoparticle contrast agents for magnetic resonance imaging is well established in clinical practice and new such contrast agents are undergoing early clinical evaluation. Likewise active (second generation) nano-therapies, exemplified by targeted control drug release systems are undergoing early clinical evaluation. The situation concerning other nano-materials such as carbon nanotubes (CNTs) and boron nitride nanotubes (BNNTs) is less advanced although considerable progress has been made on their coating for aqueous dispersion and functionalisation to enable carriage of drugs, genes and fluorescent markers. The main problem related to the clinical use of these nanotubes is that there is no consent among scientists on the fate of such nano-materials following injection or implantation in humans. Provided carbon nanotubes are manufactured to certain medical criteria (length around 1 mm, purity of 97-99% and low Fe content) they exhibit no cytotoxicity on cell cultures and demonstrate full bio-compatibility on in vivo animal studies. The results of recent experimental studies have demonstrated the potential of technologies based on CNTs for low voltage wireless electrochemotherapy of tumours and for electro-stimulation therapies for cardiac, neurodegenerative and skeletal and visceral muscle disorders.

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Background: It has recently been demonstrated that zinc oxide nanoparticles (ZnO NPs) induce death of cancerous cells whilst having no cytotoxic effect on normal cells. However, there are several issues which need to be resolved before translation of zinc oxide nanoparticles into medical use, including lack of suitable biocompatible dispersion protocols and a better understanding being needed of the mechanism of their selective cytotoxic action. \ud Methods: Nanoparticle dose affecting cell viability was evaluated in a model of proliferating cells both experimentally and mathematically. The key issue of selective toxicity of ZnO NPs toward proliferating cells was addressed by experiments using a biological model of noncancerous cells, ie, mesenchymal stem cells before and after cell differentiation to the osteogenic lineage.\ud Results: In this paper, we report a biocompatible protocol for preparation of stable aqueous solutions of monodispersed zinc oxide nanoparticles. We found that the threshold of intracellular ZnO NP concentration required to induce cell death in proliferating cells is 0.4 ± 0.02 mM. Finally, flow cytometry analysis revealed that the threshold dose of zinc oxide nanoparticles was lethal to proliferating pluripotent mesenchymal stem cells but exhibited negligible cytotoxic effects to osteogenically differentiated mesenchymal stem cells.\ud Conclusion: Results confirm the ZnO NP selective cytotoxic action on rapidly proliferating cells, whether benign or malignant

Irreversible lethal electroporation (IRE) is a new non-thermal ablation modality that usesshort pulses of high amplitude static electric fields (up 1000 V/cm) to create irreversible pores in the cell membrane, thus, causing cell death. Recently, IRE has emerged as a promising clinical modality for cancer disease treatment. Here, we investigated the responses of tumour human HeLa cells when subjected to IRE in the presence of BNNTs. These consist of tiny tubes of B and N atoms (arranged in hexagons) with diameters ranging from a 1 to 3 nanometres and lengths ,2 mm. BNNTs have attracted wide attention because of their unique electrical properties. We speculate that BNNTs, when interacting with cells exposed to static electrical fields, amplify locally the electric field, leading to cell death. In this work, electroporation assays were performed with a commercial electroporator using the cellspecific protocol suggested by the supplier (exponential decay wave, time constant 20 ms) with the specific aim to compare IRE in absence and in presence of BNNTs. We observed that BNNTs have the capacity to decrease substantially the voltage required for IRE. When cells were pulsed at 800 V/cm, we observed a 2,2-fold reduction in cell survival in the presence of BNNTs compared to controls. We conclude that the death of the tumour cells exposed to IRE is strongly enhanced in the presence of BNNTs, indicating their potential therapeutic application.