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differences in biological activity observed, given the relative similarity of physical characteristics (Table 9-1). FLG became highly cytolytic by prior association with lipids common in biomembranes (phosphatidyl choline/cholesterol). The association was relatively weak with no lipid clusters on graphene surfaces discerned by AFM after washing. The lipid-FLG association was also unstable, with eventual phase separation and agglomeration, which instability may promote penetration of cell membranes, with localised regions of high curvature and disorder in the lipid bilayer sufficient to cause the observed lysis and release of haemoglobin. Despite the interaction between the hydrocarbon region of lipids and graphene, the entry barrier of polar lipid head groups may repel pristine graphene away from lipid bilayers, which barrier may be lessened in the lytic action of GO by its edge polar regions.230, 452 The sharp edges of exfoliated pristine graphene are also proposed to slice through the polar entry barrier and allow graphene to penetrate through the hydrocarbon of lipid bilayers without causing lytic disorder in biomembranes and loss of viability.230 However, the microscopic evidence for sharp edge penetration of cell membranes was not from FLG per se. Though not well characterised, the larger flakes were first dispersed in a similar lipid (dipalmitoyl-phosphatidylcholine230), but were only considered at low graphene concentrations (~10 g/ml). Such graphene levels were only weakly cytolytic for the smaller FLG flakes used here, and may be less so for larger flakes, given lower particle concentrations for the same mass concentration. The pristine FLG here was smaller with similarly sharp edges, when any such interaction with lipid bilayers was not cytolytic, even for high concentrations of FLG with only traces of organic solvent, nor when dispersed in bile salt surfactant to facilitate entry through the polar barrier into the hydrocarbon region. Only lipid-associated FLG seriously inhibited reductive metabolism at low concentrations when tested on a breast cancer cell line (MCF-7). Though there have been reports of graphene interference with some assays452, the alamarBlue assay used here did not suffer from such underestimates of cell viability compared to other assays453, 454. However, very low concentrations of GO and FLG, particularly with traces of DMF, though not albumin nor NMP-exfoliated FLG, initially increased alamarBlue reduction in macrophages. Stimulated mitochondrial activity in response to weak uncoupling or repair Appendix 252PDF Image | PRODUCTION AND APPLICATIONS OF GRAPHENE AND ITS COMPOSITES
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