Nanoparticles traversing the extracellular matrix induce biophysical perturbation of fibronectin depicted by surface chemistry.

While the filtering and accumulation effects of the extracellular matrix (ECM) on nanoparticles (NPs) have been experimentally observed, the detailed interactions between NPs and specific biomolecules within the ECM remain poorly understood and pose challenges for in vivo molecular-level investigations. Herein, we adopt molecular dynamics simulations to elucidate the impacts of methyl-, hydroxy-, amine-, and carboxyl-modified gold NPs on the cell-binding domains of fibronectin (Fn), an indispensable component of the ECM for cell attachment and signaling. Simulation results show that NPs can specifically bind to distinct Fn domains, and the strength of these interactions depends on the physicochemical properties of NPs. NP-NH3+ exhibits the highest affinity to domains rich in acidic residues, leading to strong electrostatic interactions that induce severe deformation, potentially disrupting the normal functioning of Fn. NP-CH3 and NP-COO- selectively occupy the RGD/PHSRN motifs, which may hinder their recognition by integrins on the cell surface. Additionally, NPs can disrupt the dimerization of Fn through competing for residues at the dimer interface or by diminishing the shape complementarity between dimerized proteins. The mechanical stretching of Fn, crucial for ECM fibrillogenesis, is suppressed by NPs due to their local rigidifying effect. These results provide valuable molecular-level insights into the impacts of various NPs on the ECM, holding significant implications for advancing nanomedicine and nanosafety evaluation.

Silica nanoparticles inhibit cadmium uptake by the protozoan Tetrahymena thermophila without the need for adsorption.

The simultaneous presence of nanoparticles (NPs) and heavy metals in the environment may affect their mutual biological uptake. Although previous studies showed that NPs could alter the cellular uptake of heavy metals by their adsorption of heavy metals, whether they could affect metal uptake without the need for adsorption is unknown. This study examined the effects of silica (SiO2) NPs on the uptake of Cd ion by the protozoan Tetrahymena thermophila. We found that, even with negligible levels of adsorption, SiO2 NPs at concentrations of 3 to 100 mg/L inhibited Cd uptake. This inhibitory effect decreased as the ambient Cd concentration increased from 1 to 100 µg/L, suggesting the involvement of at least two transporters with different affinities for Cd. The transporters were subsequently identified by the specific protein inhibitors amiloride and tariquidar as NCX and ABCB1, which are responsible for the uptake of Cd at low and high Cd levels, respectively. RT-qPCR and molecular dynamics simulation further showed that the inhibitory effects of SiO2 NPs were attributable to the down-regulated expression of the genes Ncx and Abcb1, steric hindrance of Cd uptake by NCX and ABCB1, and the shrinkage of the central channel pore of the transporters in the presence of SiO2 NPs. SiO2 NPs more strongly inhibited Cd transport by NCX than by ABCB1, due to the higher binding affinity of SiO2 NPs with NCX. Overall, our study sheds new light on a previously overlooked influence of NPs on metal uptake and the responsible mechanism.

Microplastic fragmentation by rotifers in aquatic ecosystems contributes to global nanoplastic pollution.

The role of aquatic organisms in the biological fragmentation of microplastics and their contribution to global nanoplastic pollution are poorly understood. Here we present a biological fragmentation pathway that generates nanoplastics during the ingestion of microplastics by rotifers, a commonly found and globally distributed surface water zooplankton relevant for nutrient recycling. Both marine and freshwater rotifers could rapidly grind polystyrene, polyethylene and photo-aged microplastics, thus releasing smaller particulates during ingestion. Nanoindentation studies of the trophi of the rotifer chitinous mastax revealed a Young's modulus of 1.46 GPa, which was higher than the 0.79 GPa for polystyrene microparticles, suggesting a fragmentation mechanism through grinding the edges of microplastics. Marine and freshwater rotifers generated over 3.48 × 105 and 3.66 × 105 submicrometre particles per rotifer in a day, respectively, from photo-aged microplastics. Our data suggest the ubiquitous occurrence of microplastic fragmentation by different rotifer species in natural aquatic environments of both primary and secondary microplastics of various polymer compositions and provide previously unidentified insights into the fate of microplastics and the source of nanoplastics in global surface waters.

Heterogeneous aggregation of carbon and silicon nanoparticles with benzo[a]pyrene modulates their impacts on the pulmonary surfactant film.

Inhaled nanoparticles (NPs) can deposit in alveoli where they interact with the pulmonary surfactant (PS) and potentially induce toxicity. Although nano-bio interactions are influenced by the physicochemical properties of NPs, isolated NPs used in previous studies cannot accurately represent those found in atmosphere. Here we used molecular dynamics simulations to investigate the interplay between two types of NPs associated with benzo[a]pyrene (BaP) at the PS film. Silicon NPs (SiNPs), regardless of aggregation and adsorption, directly penetrated through the PS film with minimal disturbance. Meanwhile, BaPs adsorbed on SiNPs were rapidly solubilized by PS, increasing the BaP's bioaccessibility in alveoli. Carbon NPs (CNPs) showed aggregation and adsorption-dependent effects on the PS film. Compared to isolated CNPs, which extracted PS to form biomolecular coronas, aggregated CNPs caused more pronounced PS disruption, especially around irregularly shaped edges. SiNPs in mixture exacerbated the PS perturbation by piercing PS film around the site of CNP interactions. BaPs adsorbed on CNPs were less solubilized and suppressed PS extraction, but aggravated biophysical inhibition by prompting film collapse under compression. These results suggest that for proper assessment of inhalation toxicity of airborne NPs, it is imperative to consider their heterogeneous aggregation and adsorption of pollutants under atmospheric conditions.

Detection, distribution and environmental risk of metal-based nanoparticles in a coastal bay.

Metal-based nanoparticles (NPs) attract increasing concerns because of their adverse effects on aquatic ecosystems. However, their environmental concentrations and size distributions are largely unknown, especially in marine environments. In this work, environmental concentrations and risks of metal-based NPs were examined in Laizhou Bay (China) using single-particle inductively coupled plasma-mass spectrometry (sp-ICP-MS). First, separation and detection approaches of metal-based NPs were optimized for seawater and sediment samples with high recoveries of 96.7% and 76.3%, respectively. Spatial distribution results showed that Ti-based NPs had the highest average concentrations for all the 24 stations (seawater, 1.78 × 108 particles/L; sediments, 7.75 × 1012 particles/kg), followed by Zn-, Ag-, Cu-, and Au-based NPs. For all the NPs in seawater, the highest abundance occurred around the Yellow River Estuary, resulting from a huge input from Yellow River. In addition, the sizes of metal-based NPs were generally smaller in sediments than those in seawater (22, 20, 17, and 16 of 22 stations for Ag-, Cu-, Ti-, and Zn-based NPs, respectively). Based on the toxicological data of engineered NPs, predicted no-effect concentrations (PNECs) to marine species were calculated as Ag at 72.8 ng/L < ZnO at 2.66 µg/L < CuO at 7.83 µg/L < TiO2 at 72.0 µg/L, and the actual PNECs of the detected metal-based NPs may be higher due to the possible presence of natural NPs. Station 2 (around the Yellow River Estuary) was assessed as "high risk" for Ag- and Ti-based NPs with risk characterization ratio (RCR) values of 1.73 and 1.66, respectively. In addition, RCRtotal values for all the four metal-based NPs were calculated to fully assess the co-exposure environmental risk, with 1, 20, and 1 of 22 stations as "high risk", "medium risk", and "low risk", respectively. This study helps to better understand the risks of metal-based NPs in marine environments.

Molecular assembly of extracellular polymeric substances regulating aggregation of differently charged nanoplastics and subsequent interactions with bacterial membrane.

Extracellular polymeric substances (EPS) represent an interface between microbial cells and aquatic environment, where nanoplastics acquire coatings to alter their fate and toxicity. However, little is known about molecular interactions governing modification of nanoplastics at biological interfaces. Molecular dynamics simulations combining experiments were conducted to investigate assembly of EPS and its regulatory roles in the aggregation of differently charged nanoplastics and interactions with bacterial membrane. Driven by hydrophobic and electrostatic interactions, EPS formed micelle-like supramolecular structures with hydrophobic core and amphiphilic exterior. Different components, depending on their hydrophobicity and charge, were found to promote or suppress EPS assembly. Neutral and hydrophobic nanoplastics showed unbiased adsorption of EPS species, while cationic and anionic nanoplastics were distinct and attracted specific molecules of opposite charges. Compared with isolated EPS, assembled EPS concealed hydrophobic groups to be less adsorbed by nanoplastics. Aggregation of nanoplastics was alleviated by EPS due to electrostatic repulsion plus steric hindrance. ESP suppressed binding of cationic nanoplastics to the bacterial membrane through reducing the surface charge. Neutral and anionic nanoplastics showed weak membrane association, but their binding interactions were promoted by EPS. The structural details revealed here provided molecular level insights into modifications of nanoplastics at the eco-environment interface.

Dual role of pulmonary surfactant corona in modulating carbon nanotube toxicity and benzo[a]pyrene bioaccessibility.

Inhaled carbon nanotubes (CNTs) can deposit in the deep lung, where they interact with pulmonary surfactant (PS) to form coronas, potentially altering the fate and toxicity profile of CNTs. However, the presence of other contaminants in combination with CNTs may affect these interactions. Here, we used passive dosing and fluorescence-based techniques confirm the partial solubilization of BaPs adsorbed on CNTs by PS in simulated alveolar fluid. MD simulations were performed to elucidate the competition of interactions between BaPs, CNTs, and PS. We found that PS play two opposing roles in altering the toxicity profile of the CNTs. First, the formation of PS coronas reduce CNTs' toxicity by decreasing the hydrophobicity of the CNTs and decreasing their aspect ratio. Second, the interaction with PS increases the bioaccessibility of BaP through interactions with PS, which may exacerbate the inhalation toxicity of CNTs. These findings suggest that the inhalation toxicity of PS-modified CNTs should consider the bioaccessibility of coexisting contaminants, with the CNT size and aggregation state playing an important role.

Clec12a inhibits MSU-induced immune activation through lipid raft expulsion.

Monosodium uric acid (MSU) crystal, the etiological agent of gout, has been shown to trigger innate immune responses via multiple pathways. It is known that MSU-induced lipid sorting on plasma membrane promotes the phosphorylation of Syk and eventually leads to the activation of phagocytes. However, whether this membrane lipid-centric mechanism is regulated by other processes is unclear. Previous studies showed that Clec12a, a member of the C-type lectin receptor family, is reported to recognize MSU and suppresses this crystalline structure-induced immune activation. How this scenario is integrated into the lipid sorting-mediated inflammatory responses by MSU, and particularly, how Clec12a intercepts lipid raft-originated signaling cascade remains to be elucidated. Here, we found that the ITIM motif of Clec12a is dispensable for its inhibition of MSU-mediated signaling; instead, the transmembrane domain of Clec12a disrupts MSU-induced lipid raft recruitment and thus attenuates downstream signals. Single amino acid mutagenesis study showed the critical role of phenylalanine in the transmembrane region for the interactions between C-type lectin receptors and lipid rafts, which is critical for the regulation of MSU-mediated lipid sorting and phagocyte activation. Overall, our study provides new insights for the molecular mechanisms of solid particle-induced immune activation and may lead to new strategies in inflammation control.

Converting Nanotoxicity Data to Information Using Artificial Intelligence and Simulation.

Decades of nanotoxicology research have generated extensive and diverse data sets. However, data is not equal to information. The question is how to extract critical information buried in vast data streams. Here we show that artificial intelligence (AI) and molecular simulation play key roles in transforming nanotoxicity data into critical information, i.e., constructing the quantitative nanostructure (physicochemical properties)-toxicity relationships, and elucidating the toxicity-related molecular mechanisms. For AI and molecular simulation to realize their full impacts in this mission, several obstacles must be overcome. These include the paucity of high-quality nanomaterials (NMs) and standardized nanotoxicity data, the lack of model-friendly databases, the scarcity of specific and universal nanodescriptors, and the inability to simulate NMs at realistic spatial and temporal scales. This review provides a comprehensive and representative, but not exhaustive, summary of the current capability gaps and tools required to fill these formidable gaps. Specifically, we discuss the applications of AI and molecular simulation, which can address the large-scale data challenge for nanotoxicology research. The need for model-friendly nanotoxicity databases, powerful nanodescriptors, new modeling approaches, molecular mechanism analysis, and design of the next-generation NMs are also critically discussed. Finally, we provide a perspective on future trends and challenges.

Transformation, Absorption and Toxicological Mechanisms of Silver Nanoparticles in the Gastrointestinal Tract Following Oral Exposure.

Oral exposure is known as the primary way for silver nanoparticles (AgNPs), which are commonly used as food additives or antibacterial agents in commercial products, to enter the human body. Although the health risk of AgNPs has been a concern and extensively researched over the past few decades, there are still numerous knowledge gaps that need to be filled to disclose what AgNPs experience in the gastrointestinal tract (GIT) and how they cause oral toxicity. In order to gain more insight into the fate of AgNPs in the GIT, the main gastrointestinal transformation of AgNPs, including aggregation/disaggregation, oxidative dissolution, chlorination, sulfuration, and corona formation, is first described. Second, the intestinal absorption of AgNPs is presented to show how AgNPs interact with epithelial cells and cross the intestinal barrier. Then, more importantly, we make an overview of the mechanisms underlying the oral toxicity of AgNPs in light of recent advances as well as the factors affecting the nano-bio interactions in the GIT, which have rarely been thoroughly elaborated in published literature. At last, we emphatically discuss the issues that need to be addressed in the future to answer the question "How does oral exposure to AgNPs cause detrimental effects on the human body?".

A computational study of the influence of nanoparticle shape on clathrin-mediated endocytosis.

Nanoparticles have been widely used in biomedical applications such as gene/drug delivery, molecular imaging and diagnostics. Among the physicochemical properties, shape is a vital design parameter for tuning the cell uptake of nanoparticles. However, the regulatory mechanism remains elusive due to the complexity of the cell membrane and multiple pathways of cell uptake. Therefore, in this computational study, we design and clarify cell membrane wrapping on different shaped nanoparticles (sphere, rod and disk) with a clathrin assembly to model the clathrin-mediated endocytosis, which is an important pathway of nanoparticle cell uptake. Our simulations revealed that the clathrin-mediated endocytosis is shape sensitive for nanoparticles. Spherical nanoparticles are easier to be wrapped by the membrane with the self-assembly of clathrins than the other shaped nanoparticles with the same volume, and the efficiency declines with the increase in the nanoparticle shape anisotropy. Furthermore, simulation results showed clear evidence that rotation is one of the typical characteristics determining the kinetics of clathrin-mediated endocytosis of shaped nanoparticles. Especially for rod nanoparticles with high aspect ratios, nanoparticle rotation occurs in both the invagination and wrapping stages, which is different from the case without clathrins. The size and shape mismatch between the clathrin-mediated vesicle and the nanoparticle determines how the nanoparticle rotates and is wrapped by the membrane. In addition, the wrapping time of nanoparticles depends not only on the shape of the nanoparticle but also on its initial orientation and size, the rate of clathrin self-assembly and the surface tension of the membrane. These results provide insights into the interplay between cell membrane wrapping and clathrin assembly, where the nanoparticle shape matters. Understanding the dynamics mechanism of clathrin-mediated endocytosis of nanoparticles will help to design targeted nanomedicines with an improved efficacy.

Phosphatidylinositol 4,5-Bisphosphate Sensing Lipid Raft via Inter-Leaflet Coupling Regulated by Acyl Chain Length of Sphingomyelin.

Phosphatidylinositol 4,5-bisphosphate (PIP2) is an important molecule located at the inner leaflet of cell membrane, where it serves as anchoring sites for a cohort of membrane-associated molecules and as a broad-reaching signaling intermediate. The lipid raft is thought as the major platform recruiting proteins for signal transduction and also known to mediate PIP2 accumulation across the membrane. While the significance of this cross-membrane coupling is increasingly appreciated, it remains unclear whether and how PIP2 senses the dynamic change of the ordered lipid domains over the packed hydrophobic core of the bilayer. Herein, by means of molecular dynamic simulation, we reveal that inner PIP2 molecules can sense the outer lipid domain via inter-leaflet coupling, and the coupling manner is dictated by the acyl chain length of sphingomyelin (SM) partitioned to the lipid domain. Shorter SM promotes membrane domain registration, whereby PIP2 accumulates beneath the domain across the membrane. In contrast, the anti-registration is thermodynamically preferred if the lipid domain has longer SM due to the hydrophobic mismatch between the corresponding acyl chains in SM and PIP2. In this case, PIP2 is expelled by the domain with a higher diffusivity. These results provide molecular insights into the regulatory mechanism of correlation between the outer lipid domain and inner PIP2, both of which are critical components for cell signal transduction.

A zwitterionic solution for smart ionic liquids to evade cytotoxicity.

By linking the cation and anion motifs of ionic liquids (ILs), zwitterionic liquids (ZILs) exhibit at least 146-2740 and 112-1550 folds less cytotoxicity in human gastric and colon cells than those of the structurally related ILs. Computer simulation shows that ZIL molecules hardly penetrate the cell membranes in contrast to ILs. These findings reveal a novel mechanism for ZILs to evade cytotoxicity, establishing a structure-based design principle for the next generation of sustainable ZILs.

Sediment-seawater exchange altered adverse effects of ocean acidification towards marine microalgae.

Ocean acidification (OA) exhibits high threat to marine microalgae. However, the role of marine sediment in the OA-induced adverse effect towards microalgae is largely unknown. In this work, the effects of OA (pH 7.50) on the growth of individual and co-cultured microalgae (Emiliania huxleyi, Isochrysis galbana, Chlorella vulgaris, Phaeodactylum tricornutum, and Platymonas helgolandica tsingtaoensis) were systematically investigated in the sediment-seawater systems. OA inhibited E. huxleyi growth by 25.21 %, promoted P. helgolandica (tsingtaoensis) growth by 15.49 %, while did not cause any effect on the other three microalgal species in the absence of sediment. In the presence of the sediment, OA-induced growth inhibition of E. huxleyi was significantly mitigated, because the released chemicals (N, P and Fe) from seawater-sediment interface increased the photosynthesis and reduced oxidative stress. For P. tricornutum, C. vulgaris and P. helgolandica (tsingtaoensis), the growth was significantly increased in the presence of sediment in comparison with those under OA alone or normal seawater (pH 8.10). For I. galbana, the growth was inhibited when the sediment was introduced. Additionally, in the co-culturing system, C. vulgaris and P. tricornutum were the dominant species, while OA increased the proportions of dominant species and decreased the community stability as indicated by Shannon and Pielou's indexes. After the introduction of sediment, the community stability was recovered, but remained lower than that under normal condition. This work demonstrated the role of sediment in the biological responses to OA, and could be helpful for better understanding the impact of OA on marine ecosystems.

Selective membrane wrapping on differently sized nanoparticles regulated by clathrin assembly: A computational model.

Nanoparticles (NPs) enter cells via multiple pathways, all of which are NP size dependent. Previous studies indicated that the clathrin-mediated endocytosis has different selectivity for the NP size, but the regulatory mechanism remains unclear and difficult to study at the molecular scale in vivo. By means of computer simulation, here we design membrane wrapping on differently sized NPs with mimic clathrin assembly at the opposite membrane side. With relatively large NPs readily wrapped by a pure membrane as manifested, clathrin modulates the process and tunes the size selectivity. Although finite curvature can be generated by cage-like clathrin assembly to facilitate membrane wrapping on relatively small NPs, the clathrin assemblage has a certain range of size, which is mismatched with larger NPs. Besides, the local membrane patch is rigidified by clathrin to increase the barrier of membrane wrapping on larger NPs. Competition of these items determines whether membrane wrapping on NPs is promoted or suppressed, and can be tuned by the NP-membrane adhesion strength, clathrin concentration, and inter-NP distance. Our results highlight the significance of complex environment in altering the nature of NP interaction with cell membranes, and are expected to help design NPs for biomedical applications requiring precise control of NP uptake or cell membrane attachment.

Peptide Self-assembly into stable Capsid-Like nanospheres and Co-assembly with DNA to produce smart artificial viruses.

Smart artificial viruses have been successfully developed by co-assembly of de novo designed peptides with DNA, which achieved stimuli-responsibility and efficient gene transfection in cancer cells. The peptides were designed to incorporate several functional segments, including a hydrophobic aromatic segment to drive self-assembly, two or more cysteines to regulate the assemblage shape and stabilize the assembled nanostructures via forming disulfide bonds, several lysines to facilitate co-assembly with DNA and binding to cell membranes, and an enzyme-cleavable segment to introduce cancer sensitivity. The rationally designed peptides self-assembled into stable nanospheres with a uniform diameter of < 10 nm, which worked as capsid-like subunits to further interact with DNA to produce hierarchical virus-mimicking structures by encapsulating DNA in the interior. Such artificial viruses can effectively protect DNA from nuclease digestion and achieve efficient genome release by enzyme-triggered structure disassembly, which ensured a high level of gene transfection in tumor cells. The system emulates very well the structural and functional properties of natural viruses from the aspects of capsid formation, genome package and gene transfection, which is highly promising for application as efficient gene vectors.

Adsorption and catalytic degradation of preservative parabens by graphene-family nanomaterials.

Parabens pose increasing threats to human health due to endocrine disruption activity. Adsorption and degradation of parabens by three types of graphene-family nanomaterials (GFNs) were therefore investigated. For a given paraben, the maximum adsorption capacities (Q0) followed the order of reduced graphene oxide (RGO) > multilayered graphene (MG) > graphene oxide (GO); for a given GFN, Q0 followed the order of butylparaben (BuP) > propylparaben (PrP) > ethylparaben (EtP) > methylparaben (MeP), dominated by hydrophobic interaction. MeP removal by all the three GFNs was highly enhanced (0.55-4.37 times) with the assistance of H2O2 due to additional catalytic degradation process, and MG showed the highest removal enhancement. ∙OH was confirmed as the dominant radicals responsible for parabens degradation. For MG and RGO, the metal impurities (Fe, Cu, Mn, and Co) initiated Fenton-like reaction with H2O2 to generate ∙OH. GO contained oxygen-centered free radicals, which were responsible for ∙OH formation via transferring electron to H2O2. Four degradation byproducts of MeP were identified, including oxalic, propanedioic, fumaric, and 2,5-dihydroxybenzoic acids. Combined with density function theory calculations, the degradation sites and pathways were identified and confirmed. These findings provide useful information on mechanistic understanding towards the adsorption and degradation of parabens by GFNs.

Benzo[a]pyrene and heavy metal ion adsorption on nanoplastics regulated by humic acid: Cooperation/competition mechanisms revealed by molecular dynamics simulations.

Nanoplastics adsorb pollutants and organic matter to aggravate or alleviate impact to the eco-environment and human health. However, the interaction mechanisms remain unclear and difficult to study using current experimental techniques. By means of molecular dynamics simulation, here we investigate adsorption of benzo[a]pyrene (BaP) and heavy metal ions (Cu2+) on nanoplastics of different materials and surface charges regulated by humic acid (HA). Among considered materials, polystyrene shows the highest capacity of adsorbing BaPs via forming sandwiched π-stacking structures with benzene rings. Driven by hydrophobic, electrostatic and hydrogen bonding interactions, HAs spontaneously aggregate into micelle-like structures with hydrophobic core and charged exterior accessible to BaPs and Cu2+, respectively. Cationic and neutral nanoplastics adsorb more HAs to form eco-coronas, which modulate BaP and Cu2+ adsorption via following cooperation/competition mechanisms. On one hand, the direct binding of BaPs to nanoplastics is hindered by HAs through BaP encapsulation plus competitive adsorption. On the other hand, adsorbed HAs expose carboxyl groups to offer rich binding sites to promote Cu2+ adsorption on neutral and cationic nanoplastics, while unbound HAs compete with anionic nanoplastics to inhibit Cu2+ adsorption. These results provide molecular level insights into transport, transformation and accessibility of nanoplastics with coexisting contaminants in the aqueous environment.

Biosafety-inspired structural optimization of triazolium ionic liquids based on structure-toxicity relationships.

Ionic liquids (ILs), owing to their low vapor pressure and excellent solvating ability, are being increasingly applied in various industries to replace highly toxic organic solvents. They mainly pollute aquatic environment and soils, directly endangering eco-environment and human health. Therefore, it is critical to understand and optimize structural motifs of ILs with reduced toxicity. Considering human oral exposure is the major route, our investigations employed a human cell panel (modeling oral exposures) including human stomach (GES-1), intestinal (FHC), liver (HepG2) and kidney (HEK293) cells using a series of experimental and computational approaches to explore the cytotoxicity and molecular mechanism of ILs. We discovered that the cytotoxicity of triazolium and imidazolium ILs was human cell line-dependent with cytotoxicity in an order of FHC > GES-1 > HepG2 > HEK293. For this reason, a toxicity assay using a single cell line was highly inappropriate. Compared to anions (Br-, OTs-, OTMBS-) we tested, the cation of ILs played a major role in causing cytotoxicity. Ionic liquids with cations having longer hydrophobic sidechains (IL09 vs. IL01) readily insert into cell membranes with enhanced membrane and lipidomic perturbations, induce cytotoxicity by triggering cell cycle arrest and apoptosis. Reducing sidechain length and incorporating three nitrogen atoms (triazolium) instead of two (imidazolium) in the cation core alleviated cytotoxicity by reducing cell membrane perturbations and cell function interference. These findings provide important guiding principles for the design of the next-generation of "green" and safe ILs.

From reversible to irreversible: When the membrane nanotube pearling is coupled with phase separation.

Membrane nanotubes, which are ubiquitous in biology and act as channels maintaining transport between different cells and organelles, readily undergo pearling in response to external stimuli. Membrane nanotube pearling involves generation of heterogeneous curvature coupled with redistribution of membrane components that may interfere with the shape recovery of pearled nanotubes. However, the mechanism underlying such delicate process remains unclear and difficult to study at the molecular scale in vivo. By means of molecular dynamics simulation, here we investigate pearling of multi-component membrane nanotubes and reversibility through manipulating system temperature and osmotic pressure. With the equilibrium shape of membrane nanotubes controlled by the osmotic pressure, our results demonstrate that the process of membrane nanotube pearling can be reversible or irreversible, depending on the phase segregation state. For the pearled nanotube releasing high surface energy, different lipid components redistribute along the tube axial direction. Lipids with unsaturated tails prefer gathering at the high-curvature shrinking region, whereas the swelling region is constituted by saturated lipids forming the liquid-ordered phase of a higher bending rigidity. Such curvature sensitive phase segregation minimizes the system free energy by reducing both the membrane bending energy and line tension at the phase boundary. As such, the pearled nanotube fails to recover its shape upon retracting stimuli, suggesting irreversibility of the membrane nanotube pearling coupled with phase separation. Given importance of membrane nanotube pearling in various cellular activities, these results provide a new mechanism of controlling equilibrium shapes of membrane nanotubes in complex cellular environment.