Post - prandial rise of microvesicles in peripheral blood of healthy human donors

<p>Abstract</p> <p>Background</p> <p>Microvesicles isolated from body fluids are membrane - enclosed fragments of cell interior which carry information on the status of the organism. It is yet unclear how metabolism affects the number and composition of microvesicles in isolates from the peripheral blood.</p> <p>Aim</p> <p>To study the post - prandial effect on microvesicles in isolates from the peripheral blood of 21 healthy donors, in relation to blood cholesterol and blood glucose concentrations.</p> <p>Results</p> <p>The average number of microvesicles in the isolates increased 5 hours post - prandially by 52%; the increase was statistically significant (p = 0.01) with the power P = 0.68, while the average total blood cholesterol concentration, average low density lipoprotein cholesterol concentration (LDL-C) and average high density lipoprotein cholesterol concentration (HDL-C) all remained within 2% of their fasting values. We found an 11% increase in triglycerides (p = 0.12) and a 6% decrease in blood glucose (p < 0.01, P = 0.74). The post - prandial number of microvesicles negatively correlated with the post - fasting total cholesterol concentration (r = - 0.46, p = 0.035) while the difference in the number of microvesicles in the isolates between post - prandial and post - fasting states negatively correlated with the respective difference in blood glucose concentration (r = - 0.39, p = 0.05).</p> <p>Conclusions</p> <p>In a population of healthy human subjects the number of microvesicles in isolates from peripheral blood increased in the post - prandial state. The increase in the number of microvesicles was affected by the fasting concentration of cholesterol and correlated with the decrease in blood glucose.</p

Extracellular vesicle isolation yields increased by low-temperature gaseous plasma treatment of polypropylene tubes

Novel Extracellular Vesicles (EVs) based diagnostic techniques are promising non-invasive procedures for early stage disease detection which are gaining importance in the medical field. EVs are cell derived particles found in body liquids, especially blood, from which they are isolated for further analysis. However, techniques for their isolation are not fully standardized and require further improvement. Herein modification of polypropylene (PP) tubes by cold Atmospheric Pressure Plasma Jet (APPJ) is suggested to minimize the EVs to surface binding and thus increase EVs isolation yields. The influence of gaseous plasma treatment on surface morphology was studied by Atomic Force Microscopy (AFM), changes in surface wettability by measuring the Water Contact Angle (WCA), while surface chemical changes were analyzed by X-Ray Photoelectron Spectroscopy (XPS). Moreover, PP tubes from different manufacturers were compared. The final isolation yields of EVs were evaluated by flow cytometry. The results of this study suggest that gaseous plasma treatment is an intriguing technique to uniformly alter surface properties of PP tubes and improve EVs isolation yields up to 42%. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.Slovenian Research Agency (ARRS)Slovenian Research Agency - Slovenia [PR-06154

Bacterial adhesion capacity of uropathogenic Escherichia coli to polyelectrolyte multilayer coated urinary catheter surface

The application of catheters to the urinary tract is associated with nosocomial infections. Such infections are one of the most common types of infections in hospitals and health care facilities and can lead to numerous medical complications. Therefore, the understanding of the properties of urinary catheter surfaces and their potential modifications are crucial in order to reduce bacterial adhesion and subsequent biofilm formation. In our study, we consider standard polyvinyl chloride (PVC) catheter surfaces and compare their properties with the properties of the same surfaces coated with poly(diallyldimethylammonium chloride)/poly(sodium 4-styrenesulfonate) (PDADMA/PSS) polyelectrolyte multilayers. Uncoated and coated surfaces were characterized by means of roughness, hydrophobicity, and zeta potential measurements. Finally, bacterial adhesion extent of uropathogenic Escherichia coli on bare and polyelectrolyte multilayer coated surfaces was measured. The obtained results show that on non-treated surfaces, biofilm is formed which was not the case for multilayer coated surfaces. The PSS-terminated multilayer shows the lowest bacterial adhesion and could be helpful in prevention of biofilm formation. The analysis of the properties of the uncoated and coated surfaces reveals that the most significant difference is related to the charge (i.e., zeta potential) of the examined surfaces, while roughness and hydrophobicity of the examined surfaces are similar. Therefore, it could be concluded that the surface charge plays the crucial role in the bacterial adhesion on uncoated and coated PVC catheter surface

Bacterial Adhesion Capacity of Uropathogenic Escherichia coli to Polyelectrolyte Multilayer Coated Urinary Catheter Surface

The application of catheters to the urinary tract is associated with nosocomial infections. Such infections are one of the most common types of infections in hospitals and health care facilities and can lead to numerous medical complications. Therefore, the understanding of the properties of urinary catheter surfaces and their potential modifications are crucial in order to reduce bacterial adhesion and subsequent biofilm formation. In our study, we consider standard polyvinyl chloride (PVC) catheter surfaces and compare their properties with the properties of the same surfaces coated with poly(diallyldimethylammonium chloride)/poly(sodium 4-styrenesulfonate) (PDADMA/PSS) polyelectrolyte multilayers. Uncoated and coated surfaces were characterized by means of roughness, hydrophobicity, and zeta potential measurements. Finally, bacterial adhesion extent of uropathogenic Escherichia coli on bare and polyelectrolyte multilayer coated surfaces was measured. The obtained results show that on non-treated surfaces, biofilm is formed which was not the case for multilayer coated surfaces. The PSS-terminated multilayer shows the lowest bacterial adhesion and could be helpful in prevention of biofilm formation. The analysis of the properties of the uncoated and coated surfaces reveals that the most significant difference is related to the charge (i.e., zeta potential) of the examined surfaces, while roughness and hydrophobicity of the examined surfaces are similar. Therefore, it could be concluded that the surface charge plays the crucial role in the bacterial adhesion on uncoated and coated PVC catheter surfaces

Antifungal effect of polymethyl methacrylate resin base with embedded Au nanoparticles

Full and partial restorations in dentistry must replicate the characteristics of the patient’s natural teeth. Materials must have good mechanical properties and be non-toxic and biocompatible. Microbes, which can form biofilms, are constantly in contact with restorations. In this study, we investigate how well Candida albicans adheres to a polymethyl methacrylate (PMMA) resin base with gold (Au) nanoparticles. We synthesized Au nanoparticles and characterized them. The average size of Au nanoparticles embedded in PMMA was 11 nm. The color difference ∆E between PMMA and PMMA/Au composites was 2.7 and was still esthetically acceptable to patients. PMMA/Au surfaces are rougher and more hydrophilic than pure PMMA surfaces, and the isoelectric point of both types of surfaces was 4.3. Above the isoelectric point, PMMA/Au surfaces are more negatively charged than PMMA surfaces. The added Au nanoparticles decreased the tensile strength, while the hardness did not change significantly. Adhesion measurements showed that PMMA surfaces modified with Au nanoparticles reduced the extent of microbial adhesion of Candida albicans

Extracellular vesicle isolation yields increased by low-temperature gaseous plasma treatment of polypropylene tubes

Novel Extracellular Vesicles (EVs) based diagnostic techniques are promising non-invasive procedures for early stage disease detection which are gaining importance in the medical field. EVs are cell derived particles found in body liquids, especially blood, from which they are isolated for further analysis. However, techniques for their isolation are not fully standardized and require further improvement. Herein modification of polypropylene (PP) tubes by cold Atmospheric Pressure Plasma Jet (APPJ) is suggested to minimize the EVs to surface binding and thus increase EVs isolation yields. The influence of gaseous plasma treatment on surface morphology was studied by Atomic Force Microscopy (AFM), changes in surface wettability by measuring the Water Contact Angle (WCA), while surface chemical changes were analyzed by X-Ray Photoelectron Spectroscopy (XPS). Moreover, PP tubes from different manufacturers were compared. The final isolation yields of EVs were evaluated by flow cytometry. The results of this study suggest that gaseous plasma treatment is an intriguing technique to uniformly alter surface properties of PP tubes and improve EVs isolation yields up to 42%

Bacterial Adhesion Capacity of Uropathogenic Escherichia coli to Polyelectrolyte Multilayer Coated Urinary Catheter Surface

The application of catheters to the urinary tract is associated with nosocomial infections. Such infections are one of the most common types of infections in hospitals and health care facilities and can lead to numerous medical complications. Therefore, the understanding of the properties of urinary catheter surfaces and their potential modifications are crucial in order to reduce bacterial adhesion and subsequent biofilm formation. In our study, we consider standard polyvinyl chloride (PVC) catheter surfaces and compare their properties with the properties of the same surfaces coated with poly(diallyldimethylammonium chloride)/poly(sodium 4-styrenesulfonate) (PDADMA/PSS) polyelectrolyte multilayers. Uncoated and coated surfaces were characterized by means of roughness, hydrophobicity, and zeta potential measurements. Finally, bacterial adhesion extent of uropathogenic Escherichia coli on bare and polyelectrolyte multilayer coated surfaces was measured. The obtained results show that on non-treated surfaces, biofilm is formed which was not the case for multilayer coated surfaces. The PSS-terminated multilayer shows the lowest bacterial adhesion and could be helpful in prevention of biofilm formation. The analysis of the properties of the uncoated and coated surfaces reveals that the most significant difference is related to the charge (i.e., zeta potential) of the examined surfaces, while roughness and hydrophobicity of the examined surfaces are similar. Therefore, it could be concluded that the surface charge plays the crucial role in the bacterial adhesion on uncoated and coated PVC catheter surfaces

Viscosity of plasma as a key factor in assessment of extracellular vesicles by light scattering

Extracellular vesicles (EVs) isolated from biological samples are a promising material for use in medicine and technology. However, the assessment methods that would yield repeatable concentrations, sizes and compositions of the harvested material are missing. A plausible model for the description of EV isolates has not been developed. Furthermore, the identity and genesis of EVs are still obscure and the relevant parameters have not yet been identified. The purpose of this work is to better understand the mechanisms taking place during harvesting of EVs, in particular the role of viscosity of EV suspension. The EVs were harvested from blood plasma by repeated centrifugation and washing of samples. Their size and shape were assessed by using a combination of static and dynamic light scattering. The average shape parameter of the assessed particles was found to be ρ ~ 1 (0.94–1.1 in exosome standards and 0.7–1.2 in blood plasma and EV isolates), pertaining to spherical shells (spherical vesicles). This study has estimated the value of the viscosity coefficient of the medium in blood plasma to be 1.2 mPa/s. It can be concluded that light scattering could be a plausible method for the assessment of EVs upon considering that EVs are a dynamic material with a transient identity

Effect of Lidocaine and Epinephrine on Human Erythrocyte Shape and Vesiculability of Blood Cells

The effect of local anesthetic composed of lidocaine and epinephrine on vesiculability of blood cells and erythrocyte shape was studied. Whole blood and plasma were incubated with lidocaine/epinephrine. Extracellular vesicles were isolated by centrifugation and washing and counted by flow cytometry. Lidocaine/epinephrine and each component alone were added to diluted blood. Shape changes were recorded by micrographs. An ensemble of captured frames was analyzed for populations of discocytes, echinocytes, and stomatocytes by using statistical methods. Incubation of whole blood and blood plasma with lidocaine/epinephrine considerably increased concentration of extracellular vesicles in isolates (for an average factor 3.4 in blood and 2.8 in plasma). Lidocaine/epinephrine caused change of erythrocyte shape from mainly discocytic to mainly stomatocytic (higher than 50%). Lidocaine alone had even stronger stomatocytic effect (the percent of stomatocytes was higher than 95%) while epinephrine had echinocytic effect (the percent of echinocytes was higher than 80%). The differences were highly statistically significant p<10-8 with statistical power P=1. Lidocaine/epinephrine induced regions of highly anisotropically curved regions indicating that lidocaine and epinephrine interact with erythrocyte membrane. It was concluded that lidocaine/epinephrine interacts with cell membranes and increases vesiculability of blood cells in vitro