Exosomics - A Review Of Biophysics, Biology And Biochemistry Of Exosomes With A Focus On Human Breast Milk

Exosomes are biomolecular nanostructures released from cells. They carry specific biomolecular information and are mainly researched for their exquisite properties as a biomarker source and delivery system. We introduce exosomes in the context of other extracellular vesicles, describe their biophysical isolation and characterisation and discuss their biochemical profiling. Motivated by our interest in early-life nutrition and health, and corresponding studies enrolling lactating mothers and their infants, we zoom into exosomes derived from human breast milk. We argue that these should be more extensively studied at proteomic and micronutrient profiling level, because breast milk exosomes provide a more specific window into breast milk quality from an immunological (proteomics) and nutritional (micronutrient) perspective. Such enhanced breast milk exosome profiling would thereby complement and enrich the more classical whole breast milk analysis and is expected to deliver more functional insights than the rather descriptive analysis of human milk, or larger fractions thereof, such as milk fat globule membrane. We substantiate our arguments by a bioinformatic analysis of two published proteomic data sets of human breast milk exosomes

Glucose Sensor Using Redox Active Oligonucleotide-Templated Silver Nanoclusters

Redox active, photoluminescent silver nanoclusters templated with oligonucleotides were developed for glucose sensing. The silver nanoclusters had a photoluminescent emission at 610 nm that reversibly changed to 530 nm upon oxidation. The reversible emission change was measured with photoluminescent spectroscopy and used to detect H2O2, which is a by-product of the reaction of glucose with glucose oxidase. The ratio of the un-oxidised emission peak (610 nm) and the oxidised analogue (530 nm) was used to measure glucose concentrations up to 20 mM, well within glucose levels found in blood. Also, the reversibility of this system enables the silver nanoclusters to be reused

Method for the generation of surface-bound nanoparticle density gradients

We report a novel, versatile method for generating number density gradients of individual gold (Au) and silver (Ag) nanoparticles by a two-step method. First, a chemical gradient of amine surface functional groups is deposited by plasma copolymerization. Second, a density gradient of nanoparticles is formed by the immersion of the chemical gradient in solutions of nanoparticles. Chemical characterization by X-ray photoelectron spectroscopy and morphological analysis by atomic force microscopy shows that nanoparticle density closely follows the change in nitrogen surface concentration across the gradients. We also demonstrate that it is possible to control the slope of the gradients by using nanoparticle solutions of different concentrations. Important for technological and research applications, this method can be used with nanoparticles of various sizes and different materials. In addition, the use of plasma deposition allows such gradients to be generated on any type of substrate.

Aptamer Conjugated Indium Phosphide Quantum Dots with a Zinc Sulphide Shell as Photoluminescent Labels for <i>Acinetobacter baumannii</i>

Acinetobacter baumannii is a remarkable microorganism known for its diversity of habitat and its multi-drug resistance, resulting in hard-to-treat infections. Thus, a sensitive method for the identification and detection of Acinetobacter baumannii is vital. However, current methods used for the detection of pathogens have not improved in the past decades and suffer from long process times and low detection limits. A cheap, quick, and easy detection mechanism is needed. In this work, we successfully prepared indium phosphide quantum dots with a zinc sulphide shell, conjugated to a targeting aptamer ligand, to specifically label Acinetobacter baumannii. The system retained both the photophysical properties of the quantum dots and the folded structure and molecular recognition function of the aptamer, therefore successfully targeting Acinetobacter baumannii. Confocal microscopy and transmission electron microscopy showed the fluorescent quantum dots surrounding the Acinetobacter baumannii cells confirming the specificity of the aptamer conjugated to indium phosphide quantum dots with a zinc sulphide shell. Controls were undertaken with a different bacteria species, showing no binding of the aptamer conjugated quantum dots. Our strategy offers a novel method to detect bacteria and engineer a scalable platform for fluorescence detection, therefore improving current methods and allowing for better treatment

An In Vitro Investigation of Cytotoxic Effects of InP/Zns Quantum Dots with Different Surface Chemistries

Indium phosphide quantum dots (QDs) passivated with zinc sulphide in a core/shell architecture (InP/ZnS) with different surface chemistries were introduced to RAW 264.7 murine &#8220;macrophage-like&#8222; cells to understand their potential toxicities. The InP/ZnS quantum dots were conjugated with an oligonucleotide, a carboxylic acid, or an amino-polyethylene glycol ligand, and cell viability and cell proliferation were investigated via a metabolic assay. Membrane integrity was measured through the production of lactate dehydrogenase. Fluorescence microscopy showed cellular uptake. All quantum dots exhibited cytotoxic behaviour less than that observed from cadmium- or lead-based quantum dots; however, this behaviour was sensitive to the ligands used. In particular, the amino-polyethylene glycol conjugated quantum dots proved to possess the highest cytotoxicity examined here. This provides quantitative evidence that aqueous InP/ZnS quantum dots can offer a safer alternative for bioimaging or in therapeutic applications

Small surface nanotopography encourages fibroblast and osteoblast cell adhesion

In this paper, we report the initial response of 3T3 fibroblast and MG63 osteoblast cells to engineered nanotopography gradients of three nanoparticle diameters (16 nm, 38 nm and 68 nm). These nanoengineered surfaces were designed to provide a range of nanoparticle densities and comparable surface area across the gradients of different nanoparticle sizes. Importantly, we provided a uniform surface chemistry in order to be able to examine the effect of pure surface nanotopography. We found that nanotopography features of 16 nm encourage the adhesion of both cell types and that there is a critical nanoparticle density between 50 and 140 particles per μm2 where cells adhered in the greatest numbers. When nanotopography features increased to 38 nm the 3T3 cells adhered and spread well, however, the MG63 cells adhered and spread poorly. Both cell types adhered in lower numbers when the nanotopography feature size increased to 68 nm. This work demonstrates that there is a specific nanotopography scale that encourages cell adhesion and spreading, however, the preferential lateral spacing and height of the nanotopography is different for different cell types.

Interaction of silver nanoparticles with tethered bilayer lipid membranes

Silver nanoparticles are well-known for their antibacterial properties. However, the detailed mechanism describing the interaction between the nanoparticles and a cell membrane is not fully understood; Which can impede the use of the particles in biomedical applications. Here, a tethered bilayer lipid membrane has been used as a model system: to mimic a natural membrane and to. study the effect of exposure to small silver nanoparticles with diameters of about 2 nm. The solid: supported membrane architecture allowed for the: application of surface analytical techniques such as electrochemical impedance Spectroscopy and atomic force microscopy. Exposure of the membrane to solutions of the silver nanoparticles led to a small but completely reversible perturbation of the lipid bilayer

Exosomics - A Review Of Biophysics, Biology And Biochemistry Of Exosomes With A Focus On Human Breast Milk

Exosomes are biomolecular nanostructures released from cells. They carry specific biomolecular information and are mainly researched for their exquisite properties as a biomarker source and delivery system. We introduce exosomes in the context of other extracellular vesicles, describe their biophysical isolation and characterisation and discuss their biochemical profiling. Motivated by our interest in early-life nutrition and health, and corresponding studies enrolling lactating mothers and their infants, we zoom into exosomes derived from human breast milk. We argue that these should be more extensively studied at proteomic and micronutrient profiling level, because breast milk exosomes provide a more specific window into breast milk quality from an immunological (proteomics) and nutritional (micronutrient) perspective. Such enhanced breast milk exosome profiling would thereby complement and enrich the more classical whole breast milk analysis and is expected to deliver more functional insights than the rather descriptive analysis of human milk, or larger fractions thereof, such as milk fat globule membrane. We substantiate our arguments by a bioinformatic analysis of two published proteomic data sets of human breast milk exosomes

Table1.xlsx

<p>Exosomes are biomolecular nanostructures released from cells. They carry specific biomolecular information and are mainly researched for their exquisite properties as a biomarker source and delivery system. We introduce exosomes in the context of other extracellular vesicles, describe their biophysical isolation and characterisation and discuss their biochemical profiling. Motivated by our interest in early-life nutrition and health, and corresponding studies enrolling lactating mothers and their infants, we zoom into exosomes derived from human breast milk. We argue that these should be more extensively studied at proteomic and micronutrient profiling level, because breast milk exosomes provide a more specific window into breast milk quality from an immunological (proteomics) and nutritional (micronutrient) perspective. Such enhanced breast milk exosome profiling would thereby complement and enrich the more classical whole breast milk analysis and is expected to deliver more functional insights than the rather descriptive analysis of human milk, or larger fractions thereof, such as milk fat globule membrane. We substantiate our arguments by a bioinformatic analysis of two published proteomic data sets of human breast milk exosomes.</p

Table4.xlsx

<p>Exosomes are biomolecular nanostructures released from cells. They carry specific biomolecular information and are mainly researched for their exquisite properties as a biomarker source and delivery system. We introduce exosomes in the context of other extracellular vesicles, describe their biophysical isolation and characterisation and discuss their biochemical profiling. Motivated by our interest in early-life nutrition and health, and corresponding studies enrolling lactating mothers and their infants, we zoom into exosomes derived from human breast milk. We argue that these should be more extensively studied at proteomic and micronutrient profiling level, because breast milk exosomes provide a more specific window into breast milk quality from an immunological (proteomics) and nutritional (micronutrient) perspective. Such enhanced breast milk exosome profiling would thereby complement and enrich the more classical whole breast milk analysis and is expected to deliver more functional insights than the rather descriptive analysis of human milk, or larger fractions thereof, such as milk fat globule membrane. We substantiate our arguments by a bioinformatic analysis of two published proteomic data sets of human breast milk exosomes.</p