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Absorption curve and fluorescence curve (following excitation with 488 nm light) for fluorescent nanoparticles (NPs)
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The toxicology of nanomaterials is a blooming field of study, yet it is difficult to keep pace with the innovations in new materials and material applications. Those applications are quickly being introduced in research, industrial, and consumer settings. Even though the cytotoxicity of many types of nanoparticles has been demonstrated, the behavio...
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Context 1
... analysis of the fluorescent NPs showed the wave- length range in which they absorbed, which in turn revealed useful fluorescence excitation wavelengths. Figure 1 shows the absorption curve. Note the shoulder at approximately 490 nm, indicating monodispersity of NP size [24,38]. ...
Context 2
... fluorescence excitation wavelengths. Figure 1 shows the absorption curve. Note the shoulder at approximately 490 nm, indicating monodispersity of NP size [24,38]. The fluorescence was measured following excitation at a number of wavelengths (data not shown), and the emission signal following excitation with 488 nm light is shown as an example in Fig. 1. Emission after excitation at all excitation wavelengths was centered at approximately 555 nm, and the signal intensity decreased as the excitation wavelength increased. No fluorescence was detected by the fluorometer when excitation with wavelengths greater than 550 nm was used. The fluorophore excitation wavelength centered at 482 nm ...
Citations
... Certainly, it has been authenticated that lengthier periods of exposure to silica particles could improve their uptake in HepG2 cells [82]. Also, they can instigate an increment in the local dose, exposure to the product of dissolution of the exocytosis or particles and cell-to-cell particles transfer [83]. Non-porous fluorescent silica nanoparticles with different surface charges and sizes (10 nm to 200 nm) were created by Quignard et al. [84], and their prolonged contact with human dermal fibroblasts was studied. ...
Cement is utilized extensively in the manufacturing of concrete, which makes it the most common material used in building construction. However, the usage of a great deal of cement results in a great deal of CO2 emissions, which leads to the greenhouse effect. Numerous studies have developed the use of nano-SiO2 in concrete materials to lower the cement content of concrete mixtures while improving mechanical properties. Additionally, a number of studies have demonstrated that silica NPs trigger an inflammatory response in pulmonary fibroblasts. The main cells that produce and maintain the extracellular matrix (ECM) in the connection of the tissue are fibroblasts. Fibroblasts are involved in processes including tissue regeneration and wound healing. Similar to angiogenesis, inflammation, cancer, and pathological and physiological tissue fibrosis, fibroblasts act as intermediaries. The effect of silica nanoparticles on the mechanical properties of concrete (compressive strength, split tensile strength, and flexural strength) was succinctly presented in this paper. Likewise, a number of studies on the reaction of human fibroblasts to silica nanoparticles were evaluated. Numerous research on the addition of silica nanoparticles to concrete revealed that doing so significantly enhanced the material's mechanical properties. The controlled interaction of silica nanoparticles with human fibroblast cells was demonstrated to have potential in a number of applications, including aesthetics, intracellular drug release systems, improving scar tissue, determining the fate of biomaterials in vivo, and designing potential prosthetics and implant surfaces to reduce bacterial adhesion.
... The ability of the nanomaterial's to adsorb the proteins from the culture medium in the absence of cells was evaluated using PierceTM bicinchoninic acid (BCA) Protein Assay Kit (Thermo Scientific, Rockford, IL, USA). The BCA Protein Assay was performed according to the manufacturer's protocol [14]. The albumin standards (BSA) were prepared in the range suggested in protocol 0, 25, 125, 250, 500, 750, 1000, 1500, and 2000 mg/mL. ...
Magnetic nanoparticles for thermotherapy must be biocompatible and possess high thermal efficiency as heating elements. The biocompatibility of Mg 0.8 Ni 0.2 Fe 2 O 4 nanoparticles was studied using a cytotoxicity colony formation assay and a cell viability assay. HeLa cells exhibited cytotoxic effects when exposed to three different concentrations of 150 μg /ml, 100 μg /ml, and 50 μg /ml nanoparticles. Therefor e, c oncentrations of 50 μg /ml showed the lowest cytotoxic activity and the lowest toxicity to living cells. In vitro cytotoxicity of samples was then investigated by two methods, colony formation assay and cell viability assay. The Hela inhibited cell growth as 16.8% during heating by magnetic field generators.
... Wang et al. [154] measured the intracellular uptake and excretion of CuO NPs in A549 cells and found that a portion of NPs, which were located in mitochondria and the nucleus, could not be excreted by the cells. Nanoparticles that leave the endocytic vesicles or lysosomes and translocate into the cytoplasm are retained for longer periods of time and removed by exocytosis less rapidly [155]. In a typical exocytotic process, the NPs are initially trapped in lysosomes before transportation to the cell membrane for excretion. ...
Metal oxide nanoparticles (NPs) have received a great deal of attention as potential theranostic agents. Despite extensive work on a wide variety of metal oxide NPs, few chemically active metal oxide NPs have received Food and Drug Administration (FDA) clearance. The clinical translation of metal oxide NP activity, which often looks so promising in preclinical studies, has not progressed as rapidly as one might expect. The lack of FDA approval for metal oxide NPs appears to be a consequence of the complex transformation of NP chemistry as any given NP passes through multiple extra- and intracellular environments and interacts with a variety of proteins and transport processes that may degrade or transform the chemical properties of the metal oxide NP. Moreover, the translational models frequently used to study these materials do not represent the final therapeutic environment well, and studies in reduced preparations have, all too frequently, predicted fundamentally different physico-chemical properties from the biological activity observed in intact organisms. Understanding the evolving pharmacology of metal oxide NPs as they interact with biological systems is critical to establish translational test systems that effectively predict future theranostic activity.
... Attempts to directly measure attached protein by conventional determinations, as used for protein solutions, have been reported. Since protein content is measured using a UV-vis absorption method, a control for light absorbance by the suspended solid particles is needed, reducing the accuracy of the determination due to the not negligible source of interference [26]. ...
A new accurate spectrophotometric method for protein determination on nanoparticles is described. The method is based on the Coomassie blue dye that binds to the basic and aromatic amino acid residues of proteins, especially arginine and lysine. A known amount of reagent dye was mixed with a variety of protein-loaded nanoparticles. Thereafter the unconjugated reagent was mixed with excess protein (bovine serum albumin) and titrated. In this method, the reacted dye on the protein coating of nanoparticle is directly determined, in opposite to the conventional method, in which the conjugated protein is determined as the difference between the non-conjugated protein found in the supernatant after centrifugation, and the total amount of protein originally used. This method is able to measure amounts of coated protein lower than 1 ppm. •Simple and accurate method especially adapted for protein-coated nanoparticles.•The amino acid residues of protein in the nanoparticle surface react with Coomassie brilliant blue dye.•The unreacted dye is titrated with an excess of a standard protein.
... Drescher and coworkers 50 observed an agglomeration tendency of silica nanoparticles in all FCS-containing medium, the same supplement used in this study, and this condition caused a decrease in toxicity. Such behavior may be explained by the organic components of treatment media 51 , considering that the adsorption of serum proteins into silica surface can change particle compatibility, membrane contact and uptake into the cells 52 . Although micronucleus (MN) test has been considered more appropriate for evaluating the genotoxicity of the AgNP 33 , there is an issue pertaining the use of cytochalasin B in MN assay. ...
The increasing concern over the spread of diseases has lead to a high consumption of antimicrobial additives in the
medical and industrial fields. Since these particles can lixiviate from loaded materials, the contact between this
additive and mammalian cells can occur during manufacture, use and disposal of the products. Silver on fumed
silica (AgNP_SiO2) and titanium dioxide (TiO2) can be used as antimicrobial additives that are applied in polymeric
formulation. While these additives can inhibit bacteria, fungus and virus proliferation; they may also be harmful to
humans. Standard toxicological studies were undertaken using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5
diphenyltetrazolium bromide), CBPI (cytokinesis-block proliferation index) and micronucleus assay using different
sets of additive concentrations. The nanosize of the samples evaluated was confirmed by transmission electronic
microscopy. No significant micronucleus frequency increase or cell viability reduction were observed with the
exposure of L-929 murine fibroblast cells to AgNP_SiO2 and TiO2 particles at any of the tested concentrations. The
non toxic effect of the analyzed particles can be explained by considering its agglomeration tendency, composition,
and crystalline form. Further investigations should be done to understand the interference of agglomeration and
how it affects the toxicological study.
... Although a laser power of 1.5 W/cm 2 caused a certain amount of damage to the endo/lysosome, it only regulated the heat to a moderate level, which was not enough to cause extensive cell apoptosis. Since the nanoparticles that are transported from endocytotic vesicles or lysosomes to the cytoplasm have a greater difficulty undergoing exocytosis [34], the inhibition of lysosome-mediated exocytosis was attributed to the particle escape induced by moderate photothermal effect rather than cell apoptosis. ...
... and the other via a nonspecific pathway [90]. The exocytosis of nanoparticles was energy and size dependent [91][92][93][94][95]. ...
Traditional in vitro and in vivo viability assay, toxicity mechanism analysis, pathology/histopathology assays, hematology, and serum biochemistry assays have been successfully employed in nanotoxicology studies. In this chapter, the authors summarize the application of synchrotron radiation (SR) techniques in the characterization of nanomaterials as manufactured or in simulated biological systems, in the study of their in vitro and in vivo behaviors, and in the study of their ecotoxicological effects both in natural environment and in model organisms. The application of SR-based X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), small-angle X-ray scattering (SAXS), Raman spectroscopy, Fourier transformed infrared spectroscopy (FTIR), micro/nano-CT, and so on provides better resolution and sensitivity, less acquisition time, and in situ measurement compared to instruments using conventionally incident photons in the physicochemical characterization of pristine nanomaterials. Cadmium selenide quantum dots (QDs) capped with zinc sulfide have been used in the semiconductor industry and in cellular imaging.
... 54 Furthermore, nanoparticles that translocate into the cytoplasm show reduced exocytosis compared those in an endo-lysosomal pathway. 55 Activity dependent uptake of CTB-protocells ...
Cholera toxin B (CTB) modified mesoporous silica nanoparticle supported lipid bilayers (CTB-protocells) are a promising, customizable approach for targeting therapeutic cargo to motoneurons. In the present study, the endocytic mechanism and intracellular fate of CTB-protocells in motoneurons was examined to provide information for the development of therapeutic application and cargo delivery. Pharmacological inhibitors elucidated CTB-protocells endocytosis to be dependent on the integrity of lipid rafts and macropinocytosis. Using immunofluorescence techniques, live confocal and transmission electron microscopy, CTB-protocells were primarily found in the cytosol, membrane lipid domains and Golgi. There was no difference in the amount of motoneuron activity dependent uptake of CTB-protocells in neuromuscular junctions, consistent with clathrin activation at the axon terminals during low frequency activity. In conclusion, CTB-protocells uptake is mediated principally by lipid rafts and macropinocytosis. Once internalized, CTB-protocells escape lysosomal degradation, and engage biological pathways that are not readily accessible by untargeted delivery methods.
... Interestingly, in the case of bare 40 and 80 nm BPEI-AuNP, time-dependent increase in the amount of Au per cell increased up to 6 h, which then dropped and maintained an amount very similar to 3-h time point. This pattern is in line with reported literature suggesting exocytosis triggered by saturation of intracellular Au load to achieve equilibrium (Stayton et al., 2009). However, LA-and PEG-AuNP exhibited a time-dependent uptake of AuNP up to 24 h, without any numerical indications of exocytosis, possibly due to the intracellular Au load not reaching saturation threshold. ...
This study investigated the role of nanoparticle size and surface chemistry on biocorona composition and its effect on uptake, toxicity and cellular responses in human umbilical vein endothelial cells (HUVEC), employing 40 and 80 nm gold nanoparticles (AuNP) with branched polyethyleneimine (BPEI), lipoic acid (LA) and polyethylene glycol (PEG) coatings. Proteomic analysis identified 59 hard corona proteins among the various AuNP, revealing largely surface chemistry-dependent signature adsorbomes exhibiting human serum albumin (HSA) abundance. Size distribution analysis revealed the relative instability and aggregation inducing potential of bare and corona-bound BPEI-AuNP, over LA- and PEG-AuNP. Circular dichroism analysis showed surface chemistry-dependent conformational changes of proteins binding to AuNP. Time-dependent uptake of bare, plasma corona (PC) and HSA corona-bound AuNP (HSA-AuNP) showed significant reduction in uptake with PC formation. Cell viability studies demonstrated dose-dependent toxicity of BPEI-AuNP. Transcriptional profiling studies revealed 126 genes, from 13 biological pathways, to be differentially regulated by 40 nm bare and PC-bound BPEI-AuNP (PC-BPEI-AuNP). Furthermore, PC formation relieved the toxicity of cationic BPEI-AuNP by modulating expression of genes involved in DNA damage and repair, heat shock response, mitochondrial energy metabolism, oxidative stress and antioxidant response, and ER stress and unfolded protein response cascades, which were aberrantly expressed in bare BPEI-AuNP-treated cells. NP surface chemistry is shown to play the dominant role over size in determining the biocorona composition, which in turn modulate cell uptake, and biological responses, consequently defining the potential safety and efficacy of nanoformulations.
... Interestingly, in the case of bare 40 and 80 nm BPEI-AuNP, time-dependent increase in the amount of Au per cell increased up to 6 h, which then dropped and maintained an amount very similar to 3-h time point. This pattern is in line with reported literature suggesting exocytosis triggered by saturation of intracellular Au load to achieve equilibrium (Stayton et al., 2009). However, LA-and PEG-AuNP exhibited a time-dependent uptake of AuNP up to 24 h, without any numerical indications of exocytosis, possibly due to the intracellular Au load not reaching saturation threshold. ...
Gold nanoparticles (AuNP) adsorb macromolecules to form a protein corona (PC) after systemic delivery, to which the kidney as the primary excretory organ is constantly exposed. The role of the PC on AuNP cell uptake and toxicity was investigated in vitro in human proximal tubule cells (HPTC) using 40 and 80 nm branched polyethylenimine (BPEI), lipoic acid (LA) and polyethylene glycol (PEG) coated AuNP with or without (bare) PCs composed of human plasma (HP) or human serum albumin (HSA) for 0.25 to 24 h. Time-dependent intracellular uptake, assessed by ICP-MS showed PC modulated cell uptake and cytotoxicity; with bare 40 nm BPEI-AuNP showing the greatest responses. All AuNP showed minimal to no cytokine release. At the nontoxic dose, 40 nm bare BPEI-AuNP significantly modified gene expression related to immunotoxicity, steatosis, and mitochondrial metabolism; while at the high dose, pathways of DNA damage and repair, apoptosis, fatty acid metabolism and heat shock response were modulated. HP corona BPEI-AuNP response was comparable to control. These studies clearly showed reduced uptake and cytotoxicity, as well as differentiated gene expression of AuNP with PCs, questioning the utility of in vitro studies using bare NP to assess in vivo effects. Significantly, only cationic bare BPEI-AuNP had HPTC uptake or cytotoxicity suggesting the relative safety of PEG and LA-AuNP as nanomedicine constructs.
