Cellular architecture response to aspect ratio tunable nanoarrays.

Nanoarrays have been emerging as popular nanostructure platforms to investigate both cell behaviors and biological functions, due to the cell architecture respondence to the biointerface of nanostructures. Herein, we developed a series of aspect ratio tunable nanoarrays through a metal-assisted chemical etching method. Nanoarrays including nanoneedles, nanopillars, and nanoclusters were fabricated with a controllable aspect ratio. We found that nanostructures with a high aspect ratio (>10) induced significant alterations of cell physiological behaviors such as surface attachment, architecture deformation, viability, proliferation and motility. The cells on nanostructures with a high aspect ratio exhibited reorganized actin stress fibers and vimentin filaments, as well as reduced focal adhesion. This research enlarges the diversity of nanostructures on nano-bio interface investigation, provides a new insight for the surface-dependent architecture of cells, and offers unbiased understanding of factors influencing cell physiology.

Biointerface anisotropy modulates migration of breast cancer cell.

Migration of cancer cell is a cyclic process, which involves dynamic interaction between extracellular biointerface and cellular responds. In tumors, collagen as extracellular matrix reorganizes biointerface from curl and isotropic fibers to straightened and anisotropic fibers during tumorigenesis, yet how cell migration respond to topography of biointerface is unknown. In this research, we introduced a facile fabrication method on nanofibers of varying topography, which was mimicking the alignment of extracellular nanofibers, to examine the change of cytoskeleton during cell migration. We took advantage of breast carcinoma cell line (MDA-MB-231) for time-lapse imaging analysis. We found that biointerface anisotropy modulated morphology of cell and mediated the pattern of migration. Morphologically, cells on anisotropic nanofiber showed extending spindle shape. The trajectories of migration templated the topographic pattern on biointerface. Besides, aligned nanofiber induced caterpillar-like model of migration through protrusion - retraction cycle, which was indicated by periodical variation of aspect ratio and velocity of cells. The biointerface anisotropy triggered vimentin filaments and microtubule networks preferentially oriented along the alignment of nanofibers. And the velocity of cell mobility by vimentin, ß-catenin or CDC42 knockdown was significantly enhanced on aligned nanofibers. Thus, we implied that biointerface anisotropy modulated migration of breast cancer cell and it associated with reorganization of cytoskeleton filaments.

A Cuboid Spider Silk: Structure-Function Relationship and Polypeptide Signature.

A unique cuboid spider silk from the outer egg sac of Nephila pilipes, with an unusual square cross-section, is disclosed. The structure-function relationships within this silk are first studied through structural characterization, mechanical measurement, protein conformation, and polypeptide signature of silk proteins. This silk maintains the higher stiffness property of egg sac silks, and also shows a species difference. Environmental response of the mechanical properties within this silk are observed. Synchrotron FTIR microspectroscopy is used to monitor the silk protein conformation in a single natural silk. The ß-sheet structure aligns parallel to the fiber axis with a content of 22% ± 2.6%. The de novo resulting polypeptide from the solid silk fibers are novel, and an abundant polar amino acid insertion is observed. Short polyalanine (An , n ≤ 3), alternating serine and alanine (S/A)X, and alternating glycine and alanine (G/A)X, GGX, and SSX dominates in the resulting de novo polypeptide. This accords with the composition pattern of other egg sac silk proteins, besides the rarely observed GGX. This study broadens the library of egg sac spider silks and provides a new perspective to uncover structure-function relationships in spider silk.

Extracellular nanofiber-orchestrated cytoskeletal reorganization and mediated directional migration of cancer cells.

Extracellular matrix anisotropy tunes the organization and movement of surrounding cells. The primary mediators of the extracellular matrix are fiber-based materials. Natural collagen fibers reorganize from curled and isotropic fibers to straightened and anisotropic fibers during tumorigenesis, yet how the cytoskeleton is involved in the directional migration in response to the topography is unknown. To investigate this, we fabricated random, orthogonal, and aligned nanofibers to deconstruct the basic mechanisms for the migration of the human pancreatic cancer cells PANC-1 on different substrates. We found that the extracellular matrix orchestrated actin reorganization by templating the surface topography as an irregular pattern on random fibers, crossover feature on orthogonal fibers, and parallel characteristics on aligned fibers. The intermediate filament as vimentin was upregulated to form a perinuclear shape on orthogonal or aligned surfaces. We also found that the nanofiber topography mediated the directional migration via different mechanisms. The directionality ratio and velocity were statistically analyzed to unveil the pattern of directional migration. Cells on aligned nanofibers yielded a greater velocity. Rac1 and Cdc42 were involved in cell migration through regulating actin polymerization and membrane protrusions. Thus, our findings elucidate that nanofiber alignment orchestrates cytoskeletal reorganization and mediates the directional migration of cancer cells.

Internalization Characterization of Si Nanorod with Camouflaged Cell Membrane Proteins Reveals ATXN2 as a Negative Regulator.

The fabrication of shape-controlled nanocarriers is critical for efficient delivery of biomolecules across the cell membrane. Surface coating of the nanocarrier can improve internalization efficiency. Here, we developed a facile method of silicon nanorod fabrication leading to a controlled size and shape. We then systematically evaluated five surface modifications with membrane proteins from different cancer cell lines including MCF7, MD231, Hela, Panc-PDX, and Panc-1. We demonstrated that silicon nanorods coated with either a homolytic or heterolytic membrane protein coating have significantly improved internalization efficiency as compared with uncoated Si nanorods. To elucidate the molecular mechanism of the improved efficiency associated with a modified coating, we analyzed the coating membrane proteins derived from five cell lines with proteomics and identified 601 proteins shared by different cell sources. These proteins may function as cell-substrate adhesion molecules that contribute to the enhanced internalization. We also tested the internalization efficiency of nanorods with different coatings in each of the five cell lines to determine the influencing factors from target cells. We found that the internalization efficiency varied among different target cells, and the ranking of the average efficiency was as follows: Hela > Panc-PDX > MD231 > MCF7 > Panc-1. The bioinformatics analysis suggested that the low internalization efficiency in Panc-1 cells might be associated with the upregulation of ATXN2, which is a negative regulator of endocytosis. We further demonstrated that ATXN2 knockdown with specific siRNA significantly improved nanorod internalization efficiency in Panc-1 cells suggesting that ATXN2 can be a reference for efficiency prediction of nanoparticle delivery to tumor cells. Thus, we studied the effect of different cancer cell membrane proteins on nanorod uptake efficiencies. These results can improve nanorod internalization to cancer cells, including a fundamental understanding of the internalization efficiency of cancer cells.

Electrospun nanofiber regulates assembly of keratin and vimentin intermediate filaments of PANC-1 pancreatic carcinoma cells.

Intermediate filaments, together with actin microfilaments and microtubules constituent the cytoskeleton of mammalian cells, involving in various cellular activities. The roles of intermediate filaments in cell skeleton reorganization when responding with extracellular matrix (ECM) nanostructure are poorly understood yet. To unveil the effects of fibrous composition and orientation on cells, we developed electrospun nanofibers of varying topology and components, and the effects on assembly of intermediate filaments as keratin and vimentin were investigated in detail. We found that aligned nanofibers enhanced expression of E-cadherin and promoted assembly of keratin intermediate filaments. Meanwhile, the compositional variation show different preference on up-regulation of the two intermediate filaments. Compared to keratin, the assembly of vimentin intermediate filaments were promoted by incorporating bovine serum albumin (BSA) functionalized graphene oxide (BSA-GO) into polycaprolactone (PCL) nanofibers. Thus, our findings elucidate how the different physical factors of fibrous extracellular matrix affect the reorganization of cytoskeleton by assembly of keratin and vimentin intermediate filaments.

Organic-Inorganic Layered and Hollow Tin Bromide Perovskite with Tunable Broadband Emission.

Recently, layered perovskites attracted great attention for its excellent stability and light-emitting property. However, most of them rely on the toxic element lead and their emission quantum yields are generally low. Here, a unique hollow two-dimensional perovskite was developed in which the organic hexamethylene diamines (C6H18N22+) strongly coupled with distorted tin bromide anions (SnBr64-). This toxic-free low-dimensional tin perovskite exhibits a broadband emission in the visible region with a high luminescence quantum yield of 86%. First-principles calculation indicate the broadband emission is associated with the recombination of self-trapped excitons. And the emission is related to the geometry of tin bromide anions. An ultraviolet light-pumped white light emitting diode with excellent color-rendering index of 94 was fabricated using it together with a commercially available blue phosphor.

2-Thiophene ethylamine modified hyaluronic acid with its application on hepatocytes culture.

Hyaluronic acid (HA) is a component of extracellular matrix, which is important for cell functions and tissue integrity. Biosynthesized HA, as well as its derivatives, is widely used in cosmetics industry, biochemical medicine and medical surgery. In this research, we report a new hyaluronic acid derivative synthesized by amidation of hyaluronic acid with 2-thiophene ethylamine (2TEA). 2-chloro-dimethoxy-1,3,5-triazine (CDMT) served as the activating agent of the carboxylic groups. Primary mouse hepatocytes cultured with this derivative HA-2TEA maintained their epithelial morphology and showed better hepatic functions. This result was confirmed by the higher expression levels of hepatic functional genes in primary hepatocyte cultured with HA-2TEA derivative. Moreover, the protein levels of several hepatic genes were further confirmed by immunofluorescence staining. Thus HA-2TEA(2-thiopheneethylamine) derivative demonstrated good capacity on hepatocytes culture, and maintained hepatocyte functions in vitro.

Human lung epithelial cells A549 epithelial-mesenchymal transition induced by PVA/Collagen nanofiber.

Epithelial-mesenchymal transition (EMT) is a process by which epithelial cells lose their cell-cell contact to become mesenchymal stem cells, which is important on development and embryogenesis, wound healing, and cancer metastasis. This research aims to investigate the effect of topological cue as modulating factor on the EMT by tuning the diameter of electrospinning nanofiber. The cell-nanofiber interaction between human lung epithelial cell A549 and electrospinning nanofibers composed of polyvinyl alcohol (PVA) and type I collagen were investigated. The electrospinning of regenerated PVA/Collagen nanofibers were performed with water/acetic acid as a spinning solvent and glutaraldehyde as a chemical cross-linker. Parameterization on concentration, applied voltage and feeding rate was finalized to generate smooth nanofibers with good homogeneity. The scanning electron microscopy result demonstrated that A549 cell appropriately achieved extended morphology by the filopodia attaching to the surface of the nanofibrous mats. When the diameter changed from 90nm to 240nm, the A549 cell was correspondingly express varied EMT related genes. Gene expression analysis was conducted by qPCR using three typical markers for detecting EMT: N-cadherin (NCad), Vimentin (Vim), and Fibronectin (Fib). An increasing expression pattern was observed on cell culturing on 170nm sample with respect to cell cultured on 90nm and 240nm. This result indicated the 170nm PVA/Collagen nanofibers induce A549 cells to process epithelial-mesenchymal transition more seriously than those on 90nm or 240nm.

Bioinspired Conical Micropattern Modulates Cell Behaviors.

The physical properties and topological cues of a micro/nanopattern affect both cell functional activity and tissue regeneration. In this study, we developed a bioinspired approach to fabricate a microconical pattern mimicking the wing structure of beetle Coleoptera. The bioinspired micropattern with multistructural hierarchy was first screened from eight representative insect families of coleopteran. Then, we fabricated a conical micropattern through a two-step nanoimprint lithography technology. We found that human adult fibroblast cells on the bioinspired micropattern substrate had a larger spread area and higher proliferation efficiency than those on flat polydimethylsiloxane. The physical properties and topological cues affect cell functional activity. Through in situ cell motility tracking, we found that conical pillars provided specific attachment points as a mechanotransduction foundation for cell attachment to spread. The holding pillar on the bioinspired surface facilitates the actin and vimentin filament assembly into mature straightened bundles in the cytoplasm. This bioinspired substrate is transparent, flexible, and easy to fabricate. Moreover, it provides a novel in situ platform to investigate cell behaviors.

Electrospinning of PVA/sericin nanofiber and the effect on epithelial-mesenchymal transition of A549 cells.

This research aims to investigate the cell-nanomaterial interaction between epithelial-mesenchymal transition of A549 cell and electrospinning nanofibers composed of polyvinyl alcohol (PVA)/silk sericin (SS). The electrospinning of regenerated nanofiber was performed with water as a spinning solvent and glutaraldehyde as a chemical cross-linker. Solution concentration, applied voltage and spin distances as well as other parameters were optimized to generate fine nanofibers with smooth surface in good homogeneity. From the scanning electron microscopy (SEM) analysis, the nanofibers had an average diameter of 200nm. Epithelial-mesenchymal transition (EMT) is a process by which epithelial cells lose their cell polarity to become mesenchymal stem cells. This transition is affected by multiple biochemical and physical factors in cell metabolism cascade. Herein, we investigate the biophysical effect on A549 EMT by culturing cells on nanofibrous mats with different topography and composition. The cell viability was evaluated by biochemical assay and its morphology was observed with SEM. The results demonstrate that cells appropriately attached to the surface of the nanofibrous mats with extended morphology by their filopodia. Gene expression analysis was conducted by real-time PCR using multiple markers for detecting EMT: N-cadherin (NCad), Vimentin (Vim), Fibronectin (Fib) and Matrix metallopeptidase (MMP9). An increasing expression pattern was observed on NCad, Vim, Fib, with respect to a negative control as cell cultured on polystyrene dish. This result indicates the 200nm PVA/SS nanofibers may induce A549 cells to process epithelial-mesenchymal transition during the culturing.

Perovskite nanocrystals: synthesis, properties and applications.

Halide perovskites have emerged as superstar materials for optoelectronic devices. Besides the fever of research in solar cells, these materials show great promise on light emitting diodes (LEDs), photodetectors and lasers as well. Rapid advances in bulk perovskite materials aroused universal interest for the development of perovskite nanocrystals, inspired by the great progress of classic colloidal semiconductor quantum dots. Perovskite nanocrystals have been synthesized based on solution process and exhibited high luminescence quantum yield, sharp emission peak, and emission color tunability. Significant progresses have been made about the application of perovskite nanocrystals for LED and lasers in recent years. In this paper, we will comprehensively introduce the synthesis strategies, physical and chemical characteristics, as well as their applications in optoelectronic devices.