Lingna Zheng

Broad‐Spectrum Antibacterial Activity of Carbon Nanotubes to Human Gut Bacteria

Carbon nanotubes (CNTs) hold promise in manufacturing, environmental, and biomedical applications, as well as food and agricultural industries. Previous observations have shown that CNTs have antimicrobial activity; however, the impact of CNTs to human gut microbes has not been investigated. Here, the antibacterial activity of CNTs against the microbes commonly encountered in the human digestion system--L. acidophilus, B. adolescentis, E. coli, E. faecalis, and S. aureus--are evaluated. The bacteria studied include pathogenic and non-pathogenic, gram-positive and negative, and both sphere and rod strains. In this study, CNTs, including single-walled CNTs (SWCNTs, 1-3 μm), short and long multi-walled CNTs (s-MWCNTs: 0.5-2 μm; l-MWCNTs: >50 μm), and functionalized multi-walled CNTs (hydroxyl- and carboxyl-modification, 0.5-2 μm), all have broad-spectrum antibacterial effects. Notably, CNTs may selectively lyse the walls and membranes of human gut microbes, depending on not only the length and surface functional groups of CNTs, but also the shapes of the bacteria. The mechanism of antibacterial activity is associated with their diameter-dependent piercing and length-dependent wrapping on the lysis of microbial walls and membranes, inducing release of intracellular components DNA and RNA and allowing a loss of bacterial membrane potential, demonstrating complete destruction of bacteria. Thin and rigid SWCNT show more effective wall/membrane piercing on spherical bacteria than MWCNTs. Long MWCNT may wrap around gut bacteria, increasing the area making contact with the bacterial wall. This work suggests that CNTs may be broad-spectrum and efficient antibacterial agents in the gut, and selective application of CNTs could reduce the potential hazard to probiotic bacteria.

Quantitative Analysis of Gold Nanoparticles in Single Cells by Laser Ablation Inductively Coupled Plasma-Mass Spectrometry

Single cell analysis has become an important field of research in recent years reflecting the heterogeneity of cellular responses in biological systems. Here, we demonstrate a new method, based on laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS), which can quantify in situ gold nanoparticles (Au NPs) in single cells. Dried residues of picoliter droplets ejected by a commercial inkjet printer were used to simulate matrix-matched calibration standards. The gold mass in single cells exposed to 100 nM NIST Au NPs (Reference material 8012, 30 nm) for 4 h showed a log-normal distribution, ranging from 1.7 to 72 fg Au per cell, which approximately corresponds to 9 to 370 Au NPs per cell. The average result from 70 single cells (15 ± 13 fg Au per cell) was in good agreement with the result from an aqua regia digest solution of 1.2 × 10(6) cells (18 ± 1 fg Au per cell). The limit of quantification was 1.7 fg Au. This paper demonstrates the great potential of LA-ICPMS for single cell analysis and the beneficial study of biological responses to metal drugs or NPs at the single cell level.

Determination of quantum dots in single cells by inductively coupled plasma mass spectrometry

In order to assess cytotoxicity of quantum dots (QDs), new reliable analytical techniques that can provide comparative information at a single-cell level are required. In this study, a single cell ICP-MS (SC-ICP-MS) method was established to determine intracellular QDs in single cells after exposure. Uptake kinetics of QDs into cells was studied using the established method. The results were compared and validated by flow cytometry and cell digestion methods. In contrast to other methods, SC-ICP-MS can directly detect QDs and their degradation products via elements, and thus is a promising complement to available methods for single cell analysis and is expected to be a critical tool in the future.

Time-resolved ICP-MS analysis of mineral element contents and distribution patterns in single cells

Novel single cell techniques are attracting growing interest for clinical applications, because they can elucidate the cellular diversity and heterogeneity instead of the average masked by bulk measurements. Herein, time-resolved ICP-MS for the determination of essential mineral elements in single cells has been developed and is used to analyze the contents and distribution patterns of Fe, Cu, Zn, Mn, P and S in two types of cancer cells (HeLa and A549) and one type of normal cells (16HBE). The results show that there are obvious differences in contents and distribution patterns of the elements among the three types of cells. The mass of Fe, Zn, Cu, Mn, P, and S in individual HeLa cells is significantly higher and span a broader range of values than in the single 16HBE and A549 cells. The contents of Fe, Zn, and Cu follow log-normal distributions, and Mn, P, and S follow Poisson distributions with high λ values in single HeLa cells, indicating a large cell-to-cell variance. Comparatively, the contents of Cu, Zn, P, and S in 16HBE cells show the narrowest distribution range among the three tested cells, demonstrating the homogenous distribution of the elements in the cells. The method of single cell ICP-MS (SC-ICP-MS) provides potential applications for the monitoring of the variation of mineral elements at a single cell level.

Interrogating the variation of element masses and distribution patterns in single cells using ICP-MS with a high efficiency cell introduction system

AbstractCellular heterogeneity is an inherent condition of cell populations, which results from stochastic expression of genes, proteins, and metabolites. The heterogeneity of individual cells can dramatically influence cellular decision-making and cell fate. So far, our knowledge about how the variation of endogenous metals and non-metals in individual eukaryotic cells is limited. In this study, ICP-MS equipped with a high efficiency cell introduction system (HECIS) was developed as a method of single-cell ICP-MS (SC-ICP-MS). The method was applied to the single-cell analysis of Mn, Fe, Co, Cu, Zn, P, and S in human cancer cell lines (HeLa and A549) and normal human bronchial epithelial cell line (16HBE). The analysis showed obvious variation of the masses of Cu, Fe, Zn, and P in individual HeLa cells, and variation of Fe, Zn, and P in individual A549 cells. On the basis of the single-cell data, a multimodal distribution of the elements in the cell population was fitted, which showed marked differences among the various cell lines. Importantly, subpopulations of the elements were found in the cell populations, especially in the HeLa cancer cells. This study demonstrates that SC-ICP-MS is able to unravel the extent of variation of endogenous elements in individual cells, which will help to improve our fundamental understanding of cellular biology and reveal novel insights into human biology and medicine. Graphical abstractThe variations of masses and distribution patterns of elements Mn, Fe, Co, Cu, Zn, P, and S in single cells were successfully detected by ICP-MS coupled with a high efficiency cell introduction system (HECIS)

Coculture with Low-Dose SWCNT Attenuates Bacterial Invasion and Inflammation in Human Enterocyte-like Caco-2 Cells

Single walled carbon nanotubes (SWCNTs) have been shown to be highly effective against a wide range of bacteria. Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) infection is a well-known mediator to prolong hospitalization and initiate chronic inflammation, yet the biological effects of SWCNTs on the pathogen-infected enterocytes remain unclear. Herein, it is shown that the low-dose SWCNT treatment attenuates the human enterocyte-like Caco-2 cells from the damage of E. coli and S. aureus infection by suppressing NLRP3 inflammasome activation. The relatively low-dose (1 and 10 μg mL(-1) ) SWCNT treatments reduce the adhesion and invasion of E. coli and S. aureus to Caco-2 cells, increase the cell viability and proliferation, reduce the tight junction permeability, and restitute the integrity of cell surface microvilli structure, meanwhile has low cytotoxicity to the host cells. The low-dose SWCNT treatment further reduces the NLRP3-mediated IL-1β secretion in the infected cells. The results identify that a low-dose SWCNT treatment serves a protective function for the E. coli- and S. aureus-infected Caco-2 cells by negatively regulating mitochondrial reactive oxygen species-mediated NLRP3 inflammasome activation.

Peptide–Au Cluster Probe: Precisely Detecting Epidermal Growth Factor Receptor of Three Tumor Cell Lines at a Single-Cell Level

Alterations in protein (e.g., biomarkers) expression levels have a significant correlation with tumor development and prognosis; therefore, it is desired to develop precise methods to differentiate the expression level of proteins in tumor cell lines, especially at the single-cell level. Here, we report a precise and versatile approach of quantifying the protein expression levels of three tumor cell lines in situ using a peptide–Au cluster probe. The probe (Au5Peptide3) consists of a peptide with a specific cell membrane epidermal growth factor receptor (EGFR) targeting ability and an Au cluster for both cell membrane EGFR imaging using confocal microscopy and cell membrane EGFR counting by laser ablation inductively coupled plasma mass spectrometry. Utilizing the peptide–Au cluster probe, we successfully quantify the EGFR expression levels of SMMC-7721, KB, and HeLa cells at a single-cell level and differentiate the EGFR expression levels among these cell lines. The peptide–Au cluster probe, with the ability to differentiate the protein expression level of different cell lines, shows exceptional promise for providing reliable predictive and prognostic information of tumors at a single-cell level.

Elemental Bio-imaging of Biological Samples by Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry

Abstract Trace elements play very important roles in biological organism and are closely related to many diseases. New analytical methods are urgently needed for in situ determination of trace elements in biological tissues or single cells. A method based on laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) was developed here for in situ analysis. Line ablation mode was chosen and the output energy of the laser was optimized at a low fluence of less than 1 J cm −2 . Two kinds of elemental image of both a coronal section from mouse brain and single cells exposed to gold nanoparticles were obtained by the developed method. Due to the unique characteristic, such as good spatial resolution, excellent detection limit, and reasonable running cost, LA-ICP-MS will be applied widely and become a useful tool in biomedical research in the future.

Inhibition of Lysozyme Fibrillation by Gold Nanorods and Nanoparticles

Amyloid fibrillation has been implicated in many neurodegenerations, dialysis-related amyloidosis, type II diabetes and more than 30 other amyloid-related diseases. Nanomaterials as potential inhibitors of amyloid fibrillation have attracted increasing interests. In the present study, the effects of gold nanorods (AuNRs) and nanoparticles (AuNPs) on amyloid fibrillation were investigated using hen egg white lysozyme (HEWL) as a model system. Our results indicated that AuNRs and AuNPs, especially AuNRs, present significant inhibitory effects on HEWL amyloid fibril formation during all the kinetic processes, from nucleation to elongation and equilibration stages. The stronger adsorption capacity of HEWL on AuNRs surface is the key mechanism of inhibition of HEWL amyloid fibrillation. Furthermore, AuNRs lead to more stable α-helix conformation and hydrophobic microenvironment of aromatic side groups in HEWL molecules, which facilitate the system to form small amorphous aggregates rather than oligomer, profibril or mature fibril.