Use of Nanosphere Self-Assembly to Pattern Nanoporous Membranes for the Study of Extracellular Vesicles.

Nanoscale biocomponents naturally released by cells, such as extracellular vesicles (EVs), have recently gained interest due to their therapeutic and diagnostic potential. Membrane based isolation and co-culture systems have been utilized in an effort to study EVs and their effects. Nevertheless, improved platforms for the study of small EVs are still needed. Suitable membranes, for isolation and co-culture systems, require pore sizes to reach into the nanoscale. These pore sizes cannot be achieved through traditional lithographic techniques and conventional thick nanoporous membranes commonly exhibit low permeability. Here we utilized nanospheres, similar in size and shape to the targeted small EVs, as patterning features for the fabrication of freestanding SiN membranes (120 nm thick) released in minutes through a sacrificial ZnO layer. We evaluated the feasibility of separating subpopulation of EVs based on size using these membranes. The membrane used here showed an effective size cut-off of 300 nm with the majority of the EVs ≤200 nm. This work provides a convenient platform with great potential for studying subpopulations of EVs.

Enzymatic Biotransformation of Pomegranate Ellagitannins: Initial Approach to Reaction Conditions.

BACKGROUND: Ellagitannase (Ellagitannin acyl hydrolase) is an inducible enzyme with great potential use in food industry since allows the ellagic acid release from ellagitannins. OBJECTIVE: In this work, ellagitannase was produced by the fungus Aspergillus niger GH1 in solid state fermentation using polyurethane foam as solid support and pomegranate husk ellagitannins as sole carbon source and ellagitannase inducer and an initial approach to the enzymatic reaction conditions was reached. MATERIALS AND METHODS: Ellagitannase was produced by Aspergillus niger GH1 in solid state fermentation and the ideal reaction conditions for ellagitannase activity based on ellagic acid quantification as ellagitannins biotransformation product by high performance liquid chromatographic are reported. RESULTS: The enzyme ideal reaction conditions were substrate concentration of 1 mg.mL-1, 60 °C and pH 5.0, during 10 min of reaction. The kinetic enzyme constants (V max = 30.34 mM.mL-1.min-1 and K m = 1.48 x 103 mM) using punicalagin assubstrate were determined. CONCLUSION: The assay was completed in a short time and may find application in future studies of ellagic acid production.

Use of porous membranes in tissue barrier and co-culture models.

Porous membranes enable the partitioning of cellular microenvironments in vitro, while still allowing physical and biochemical crosstalk between cells, a feature that is often necessary for recapitulating physiological functions. This article provides an overview of the different membranes used in tissue barrier and cellular co-culture models with a focus on experimental design and control of these systems. Specifically, we discuss how the structural, mechanical, chemical, and even the optical and transport properties of different membranes bestow specific advantages and disadvantages through the context of physiological relevance. This review also explores how membrane pore properties affect perfusion and solute permeability by developing an analytical framework to guide the design and use of tissue barrier or co-culture models. Ultimately, this review offers insight into the important aspects one must consider when using porous membranes in tissue barrier and lab-on-a-chip applications.

Fabrication techniques enabling ultrathin nanostructured membranes for separations.

The fabrication of nanostructured materials is an area of continuous improvement and innovative techniques that fulfill the demand of many fields of research and development. The continuously decreasing size of the smallest patternable feature has expanded the catalog of methods enabling the fabrication of nanostructured materials. Several of these nanofabrication techniques have sprouted from applications requiring nanoporous membranes such as molecular separations, cell culture, and plasmonics. This review summarizes methods that successfully produce through-pores in ultrathin films exhibiting an approximate pore size to thickness ratio of one, which has been shown to be beneficial due to high permeability and improved separation potential. The material reviewed includes large-area, parallel, and affordable approaches such as self-organizing polymers, nanosphere lithography, anodization, nanoimprint lithography as well as others such as solid phase crystallization and nanosphere lens lithography. The aim of this review is to provide a set of inexpensive fabrication techniques to produce nanostructured materials exhibiting pores ranging from 10 to 350 nm and a pore size to thickness ratio close to one. The fabrication methods described in this work have reported the successful manufacture of nanoporous membranes exhibiting the ideal characteristics to improve selectivity and permeability when applied as separation media in ultrafiltration.

Nitroxide-Functionalized Graphene Oxide from Graphite Oxide.

A facile method for preparing functionalized graphene oxide single layers with nitroxide groups is reported herein. Highly oxidized graphite oxide (GO=90.6%) was obtained, slightly modifying an improved Hummer's method. Oxoammonium salts (OS) were investigated to introduce nitroxide groups to GO, resulting in a one-step functionalization and exfoliation. The mechanisms of functionalization/exfoliation are proposed, where the oxidation of aromatic alcohols to ketone groups, and the formation of alkoxyamine species are suggested. Two kinds of functionalized graphene oxide layers (GOFT1 and GOFT2) were obtained by controlling the amount of OS added. GOFT1 and GOFT2 exhibited a high interlayer spacing (d0001 = 1.12nm), which was determined by X-ray diffraction. The presence of new chemical bonds C-N (~9.5 %) and O-O (~4.3 %) from nitroxide attached onto graphene layers were observed by X-ray photoelectron spectroscopy. Single-layers of GOFT1 were observed by HRTEM, exhibiting amorphous and crystalline zones at a 50:50 ratio; in contrast, layers of GOFT2 exhibited a fully amorphous surface. Fingerprint of GOFT1 single layers was obtained by electron diffraction at several tilts. Finally, the potential use of these materials within Nylon 6 matrices was investigated, where an unusual simultaneous increase in tensile stress, tensile strain and Young's modulus was observed.