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The laser surface modification of metallic implants presents a promising alternative to other surface modification techniques. A total of four alloyed metallic biomaterials were used for this study: medical steel (AISI 316L), cobalt–chromium–molybdenum alloy (CoCrMo) and titanium alloys (Ti6Al4V and Ti6Al7Nb). Samples of metallic biomaterials after machining were subjected to polishing or laser modification in two different versions. The results of surface modification were documented using SEM imaging and roughness measurement. After modification, the samples were sterilized with dry hot air, then exposed to citrate blood, washed with PBS buffer, fixed with glutaraldehyde, sputtered with a layer of gold and imaged using SEM to enable the quantification of adhered, activated and aggregated platelets on the surface of biomaterial samples. The average total number, counted in the field of view, of adhered platelets on the surfaces of the four tested biomaterials, regardless of the type of modification, did not differ statistically significantly (66 ± 81, 67 ± 75, 61 ± 70 and 57 ± 61 for AISI 316L, CoCrMo, Ti6Al4V and Ti6Al7Nb, respectively) and the average number of platelet aggregates was statistically significantly higher (p < 0.01) on the surfaces of AISI 316L medical steel (42 ± 53) and of the CoCrMo alloy (42 ± 52) compared to the surfaces of the titanium alloys Ti6Al4V (33 ± 39) and Ti6Al7Nb (32 ± 37). Remaining blood after contact was used to assess spontaneous platelet activation and aggregation in whole blood by flow cytometry. An in-depth analysis conducted on the obtained results as a function of the type of modification indicates small but statistically significant differences in the interaction of platelets with the tested surfaces of metallic biomaterials. [ABSTRACT FROM AUTHOR]

Understanding of biology of osteosarcoma malignant progression is indispensable for enhancement of conventional chemotherapy by the use of silver nanoparticles (AgNPs). We presented an in vitro model of cancer progression closely resembling processes occurring in vivo in terms of protein profile. A comparison of cytotoxic and genotoxic potential of AgNPs in Saos‐2 cells in early stages of cancerous progression (early passages) with the cells in advanced stages (late passages) demonstrated significantly reduced responsiveness of the late passage cells to nanoparticles toxicity. It was also confirmed by proteome analysis as we identified considerably higher number of differentially expressed proteins in Saos‐2 cells in early passages compared to the late passage cells. Our studies showed that the ability of AgNPs as potential drug carriers to deliver a medication and/or to evoke toxic effects might be significantly diminished in advanced stages of cancer progression. [ABSTRACT FROM AUTHOR]

The aim of this study is to assess the effect of PAMAM dendrimers of second, fourth, and seventh generations on human umbilical vein endothelial cells. Primary endothelial cells were exposed to PAMAM dendrimers for 24 h, using concentrations reducing cellular viability to the levels of 90, 75, and 50%. We assumed, that changes in mechanical properties reflect toxicity of PAMAM dendrimers. The mechanical properties were investigated using atomic force spectroscopy (AFS) technique with the use of two approaches for measuring cell elasticity: global, where the tests were performed using a micrometer-hemispherical probe, and local, where a nanometer-sized probe was used. For the sharp probe, a reduction in the elasticity modulus was observed in comparison to untreated control cells, that is related to the depolymerization of the cytoskeleton and the processes leading to cell apoptosis. In the case of the hemispherical probe, cell softening was also observed in comparison to control cells, but with increasing PAMAM concentrations, the modulus of elasticity increases. It is related to the sensing of numerous intracellular vesicles with the use of this probe, e.g. endosomal and empty plasmalemmal which can also alter cell elasticity. The presence of external and intracellular vesicles was confirmed by scanning and transmission electron microscopy. The relationship between the elasticity of HUVEC cells exposed to PAMAM dendrimers of selected generations and their toxic effects was presented herein for the first time. In the transmission electron microscopy images of the cells exposed to PAMAM dendrimers, we have also observed distinctive vesicles with regular multilayer arranged structure. [ABSTRACT FROM AUTHOR]

Research background. Cellulose is an ingredient of waste materials that can be converted to other valuable substances. This is possible provided that the polymer molecule is degraded to smaller particles and used as a carbon source by microorganisms. Because of the frequently applied methods of pretreatment of lignocellulosic materials, the cellulases derived from thermophilic microorganisms are particularly desirable. Experimental approach. We were looking for cellulolytic microorganisms able to grow at 50 °C and we described their morphological features and biochemical characteristics based on carboxymethyl cellulase (CMCase) activity and the API® ZYM system. The growth curves during incubation at 50 °C were examined using the BioLector® microbioreactor. Results and conclusions. Forty bacterial strains were isolated from fermenting hay, geothermal karst spring, hot spring and geothermal pond at 50 °C. The vast majority of the bacteria were Gram-positive and rod-shaped with the maximum growth temperature of at least 50 °C. We also demonstrated a large diversity of biochemical characteristics among the microorganisms. The CMCase activity was confirmed in 27 strains. Hydrolysis capacities were significant in bacterial strains: BBLN1, BSO6, BSO10, BSO13 and BSO14, and reached 2.74, 1.62, 1.30, 1.38 and 8.02 respectively. Rapid and stable growth was observed, among others, for BBLN1, BSO10, BSO13 and BSO14. The strains fulfilled the selection conditions and were identified based on the 16S rDNA sequences. BBLN1, BSO10, BSO13 were classified as Bacillus licheniformis, whereas BSO14 as Paenibacillus lactis. Novelty and scientific contribution. We described cellulolytic activity and biochemical characteristics of many bacteria isolated from hot environments. We are also the first to report the cellulolytic activity of thermotolerant P. lactis. Described strains can be a source of new thermostable cellulases, which are extremely desirable in various branches of circular bioeconomy. [ABSTRACT FROM AUTHOR]

This publication describes analytical data for four psychoactive substances: 5F-PB-22, NM-2201, UR-144 and AB-CHMINACA. Compounds were synthesized in our laboratory and their presence was confirmed using electrospray ionisation mass spectrometry (ESI-MS), gas chromatography with electron ionization mass spectrometry detection (GC-EI-MS), gas chromatography with flame ionization detection (GC-FID), high-performance liquid chromatography (HPLC), Raman spectroscopy as well as proton nuclear magnetic resonance (1H NMR). Mass spectra show molecular ions and adducts peaks. Characteristic fragmentation patterns for cannabinoids can be seen by referring to GC-EI-MS mass spectra. Simple and sensitive HPLC methods were developed for the qualitative and quantitative determination of all synthesized compounds. RAMAN and GC-FID served as analytical tools for compound identification in the stability test. [ABSTRACT FROM AUTHOR]

Endothelial cell aging is related to changes not only in cell phenotype, such as luminal changes, intimal and medial thickening, and increased vascular stiffness, but encompasses different cell responses to various substances including drugs or nanomaterials. In the present work, time‐ and dose‐dependent elasticity changes evoked by silver nanoparticles in endothelial cells in early (below 15) passages were analyzed. Silver nanoparticle concentrations of 3, 3.6, and 16 μg/mL were selected for elasticity measurements for long incubation (24 hours) and of 1 and 3 μg/mL for monitoring dynamic elasticity changes of 1‐, 3‐, and 6‐hour incubations. Surprisingly, a significant reduction in the cells elasticity modulus at lower number of passages exposed to silver nanoparticles used at 3 μg/mL for 24 hours was demonstrated. These results are in contrast to those obtained for endothelial cells in late (33‐43) passages that may result from cellular aging in response to nanosilver. Furthermore, for short incubation times (1 and 3 hours), SNP‐induced significant increase in the cell elasticity modulus was detected. In current work, we also attempted to answer the question whether the changes in cell elasticity were induced by the silver nanoparticles stabilized with polyvinyl pyrrolidone or by stabilizer itself. Elasticity measurements were supplemented by observations made with transmission electron microscopy and scanning electron microscopy, which confirmed the presence of silver nanoparticles inside the cells and on the cell membrane. Additionally, activation of reactive oxygen species was detected for cells exposed to SNPs for 1 and 3 hours, which was accompanied by increased cell elasticity modulus suggesting a possible mechanism of observed phenomenon. [ABSTRACT FROM AUTHOR]

The hybrid technology combines an efficient material‐removal process and implant surface treatment by the laser reducing time of manufacture process compared to currently used machining technologies. It also permits precise structuring of the implant material surface. Six structures of the Ti6Al4V ELI surface were designed and studied how the structure topography prepared with the hybrid technology affected the Escherichia coli adhesion to the surface and viability, as well as the growth, adhesion, and viability of human osteogenic Saos‐2 cells cultured on the investigated surfaces. Results have confirmed that the microtopography of medical titanium alloy plays a beneficial role in bacterial adhesion and viability (number of bacteria found on reference surface: [5.9 ± 0.44] × 106 CFU/ml, sample no. 3: [8.8 ± 0.93] × 104 CFU/ml, and sample no. 5: [1.2 ± 0.23] × 107 CFU/ml; CFU ‐ Colony Forming Unit). All tested structured surfaces enabled good cell attachment and proliferation of Saos‐2 cells (viability of Saos‐2 cells [% of control] for reference surface: 81.93%; sample no. 3: 75% and sample no. 5: 100%). Transcriptome analysis of genes commonly expressed in the process of osseointegration demonstrated that the use of hybrid technology allows designing structures that enhance osseointegration but it should be coupled with other methods of preventing bacterial growth, or with a different strategy to limit microbial colonization with the satisfactory osseointegration potential. [ABSTRACT FROM AUTHOR]

Today, the extensive and constantly growing number of applications in the field of nanotechnology poses a lot of questions about the potential toxicity of nanomaterials (NMs) toward cells of different origins. In our work we employed the tools of molecular biology to evaluate changes that occur in human endothelial cells at the transcriptomic and proteomic level, following 24 h of exposure to three different classes of NMs. Using microarray technology, we demonstrated that 24 h of exposure to silver nanoparticles (SNPs), multiwalled carbon nanotubes (MWCNTs) and polyamidoamine dendrimers (PAMAMs) leads to changes in 299, 1271, and 431 genes, respectively, influencing specific molecular pathways. The 2D‐DIGE and mass spectrometry analysis revealed that differentially expressed proteins were involved in numerous cellular processes, for example, cytoskeletal reorganization, cell growth and proliferation, or response to stress. Both, transcriptome and proteome alterations indicate reorganization of mechanism regulating cell functioning. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1024–1034, 2019. [ABSTRACT FROM AUTHOR]

Abstract: Endothelial cells, due to their location, are interesting objects for atomic force spectroscopy study. They constitute a barrier between blood and vessel tissues located deeper, and therefore they are the first line of contact with various substances present in blood, eg, drugs or nanoparticles. This work intends to verify whether the mechanical response of immortalized human umbilical vein endothelial cells (EA.hy926), when exposed to silver nanoparticles, as measured using force spectroscopy, could be effectively used as a bio‐indicator of the physiological state of the cells. Silver nanoparticles were characterized with transmission electron microscopy and dynamic light scattering techniques. Tetrazolium salt reduction test was used to determine cell viability after treatment with silver nanoparticles. An elasticity of native cells was examined in the Hanks' buffer whereas fixed cells were softly fixed with formaldehyde. Additional aspect of the work is the comparative force spectroscopy utilizing AFM probes of ball‐shape and conical geometries, in order to understand what changes in cell elasticity, caused by SNPs, were detectable with each probe. As a supplement to elasticity studies, cell morphology observation by atomic force microscopy and detection of silver nanoparticles inside cells using transmission electron microscopy were also performed. Cells exposed to silver nanoparticles at the highest selected concentrations (3.6 μg/mL, 16 μg/mL) are less elastic. It may be associated with the reorganization of the cellular cytoskeleton and the “strengthening” of the cell cortex caused by presence of silver nanoparticles. This observation does not depend on cell fixation. Agglomerates of silver nanoparticles were observed on the cell membrane as well as inside the cells. [ABSTRACT FROM AUTHOR]

ABSTRACT: In this article, the properties of thermosensitive chitosan hydrogels (structural and biological) prepared using chitosan chloride and β‐glycerophosphate are presented. The porous structure of the gels was observed with a scanning electron microscopy (SEM). Rheological and other structural and morphological investigations were carried out by infrared spectroscopy and X‐ray photoelectron spectroscopy. Crystallinity of the gel was determined by X‐ray diffraction analysis. Biocompatibility of the chitosan gels was investigated by contact with bone cells (line Saos‐2) for 7 and 21 days. The cells were encapsulated and seeded onto the gel. A live/dead test showed no cytotoxic reaction. The structure of the gels, before and after the cell culture, was tested using SEM (including ESEM), to select the most suitable method of preparation and reliable visualization of the gel with cells. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018, 135, 46459. [ABSTRACT FROM AUTHOR]

Nematode Caenorhabditis elegans (C. elegans) was used to investigate the impact of silver nanoparticles (SNP), multiwalled carbon nanotubes (MWCNT), and polyamidoamine dendrimers (PAMAM) used in concentration of 1010 particle/mL. Population-based observations and gene expression analysis were employed in this study. SNP and PAMAM caused decrease in the number of live nematodes and their body length, but MWCNT did not affect the population of nematodes. Gene expression analysis revealed significant changes caused by the presence of all studied nanomaterials, and the results strongly suggest a specific metabolic response of the nematode organism to exposure to various nanomaterials. It was shown that C. elegans is a very sensitive organism capable to respond specifically to the exposure to some nanomaterials and therefore could be considered as a possible biosensor for early warning of presence of some nanoparticles. [ABSTRACT FROM AUTHOR]

The aim of present study was to determine the hemocompatibility, cellular response of endothelial cells and bacterial adhesion to a new polyester nanocomposite. The carbon nanoparticle nanocomposite was prepared via in situ polymerization of monomers to obtain material of hardness 55 Sh D similar to polyurethanes used in medical applications, for example, in heart-assisting devices. The carbon nanoparticle-containing polyester exhibits markedly reduced bacterial colonization, as compared to commercially available polyurethanes. Further the nanocomposite possesses markedly improved hemocompatibility, as determined by flow cytometry, and robust endothelialization. Possible explanations for these beneficial properties include surface nanoroughness of carbon nanoparticle-containing nanocomposites and presence of fatty acid sequences within polymer structure. [ABSTRACT FROM AUTHOR]

In this article, properties of thermosensitive chitosan hydrogels prepared with the use of chitosan chloride with β-glycerophosphate disodium salt pentahydrate enriched with calcium glycerophosphate are presented and compared with chitosan hydrogels with β-glycerophosphate disodium salt pentahydrate. The study is focused on the determination of hydrogel structure and biological testing of hydrogels with human osteoblasts line Saos-2. The structure of gels was visualized by scanning electron microscopy and was investigated by infrared spectroscopy. The crystallinity of gel structure was determined by X-ray diffraction analysis and thermal effects were determined using differential scanning calorimetry thermograms. [ABSTRACT FROM AUTHOR]

Most biomaterials at present have sufficient mechanical properties; however compliance with standards for biocompatibility is often not sufficient in clinical practice. This may be due to the complexity of biological systems in general and the diversity of individual responses to these materials by implant recipients. Significant improvement of biocompatibility must involve surface modification of implants, which in the future will make it possible to introduce individually selected types of surface modification for individual recipients. The key to this technology seems to be understanding the processes occurring at the site of contact of the implant with the tissue. Processes resulting from the stress generated by the contact of the biomaterial surfaces were observed with endothelial cells line EA.hy926, and it was demonstrated that differently modified surfaces of medical steel (polished medical steel and medical steel coated with Parylene C and nanocrystalline diamond) cause diverse cellular response in cells grown on these surfaces, on both the cellular (cell morphology and cell survival) and molecular (transcriptome and proteome profiles) levels. The herein presented observations are a good starting point not only for further research and the development of far-reaching personalization of medical implants, but also to study the potential use of cells as a specific sensor capable of recognizing different surfaces with which these cells come into contact. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 880-893, 2016. [ABSTRACT FROM AUTHOR]

The aim of the present work was to examine the interactions of parylene C with such selected biological objects as: blood plasma proteins, platelets, endothelial cells, and bacterial biofilm produced by E. coli cells. The results obtained strongly support the thesis that parylene C is a material worth considering for biomedical use. Parylene C coating on polished medical steel significantly reduces platelet adhesion to this surface. On the other hand, in the case of the surface of machined medical steel coated with parylene C, the number of adhered platelets is significantly higher. This also means that surface texture of substrate material is very well reproduced by parylene C coating and is an important factor facilitating the platelet adhesion. Adsorption of plasma proteins at parylene C surface is very effective, and this finding confirms a notion that cell interaction with surfaces is mediated by the adsorbed proteins. In the light of the above, a high susceptibility of parylene C surface to bacterial colonization is easy to explain. The results showing reduced proliferation and changes in endothelial cell gene expression should also be seriously analysed when parylene C is considered for the use in contact with blood vessels. [ABSTRACT FROM AUTHOR]

Microscopic methods usable for sample surface imaging and subsequent qualitative and quantitative evaluation of platelet adhesion to the surface of the biomaterial studied were compared. It was shown, making use of the samples of medical steel (AISI 316L), that such tools as surface imaging with scanning electron microscopy (SEM), glutaraldehyde induced fluorescence technique (GIFT) and metallurgical microscopy (MM) are equivalent in evaluating surface platelet adhesion. The importance of biological variability of blood samples for a proper result assessment and the necessity of using internal standards were also considered. [ABSTRACT FROM AUTHOR]

Silicone-urethanes, polymers combining the characteristics of two widely used biomaterials, i.e. polyurethanes and silicones, are highly valued in many applications, including medical implants. To assess properties of these materials in contact with living cells, a set of different silicone-urethane materials, candidates for tissue engineering scaffolds, was synthesized and characterized. Two different oligomeric siloxane diols: Tegomer-2111 (Teg) and KF-6001 (KF), and two different types of diisocyanate, MDI and IPDI, were used in synthesis. Blood platelets adhesion to surfaces of selected materials showed a higher thrombogenicity of material based on Teg. Human fibroblasts were used in in vitro biocompatibility tests. The viability of cells cultured on silicone-urethanes was tested by XTT assay. Teg-based silicone-urethanes showed a significantly higher biocompatibility than those based on KF. Materials based on MDI compared to IPDI were found to be significantly more favoured by cells, not necessarily due to the type of diisocyanate but maybe also because of the necessity of using potentially toxic catalyst which accompanies the use of IPDI. Our studies indicate that silicone-urethanes are potent materials for tissue engineering products development. On the basis of the observations performed in cell culture, Tegomer- 2111 as oligomeric siloxane diol and MDI as diisocyanate are recommended as starting materials for silicone-urethane scaffolds synthesis. [ABSTRACT FROM AUTHOR]

The article presents abstracts of research about platelets. They include "Insulin signaling in diabetes mellitus type 2 platelets," "Role for the F11 receptor (F11R, JAM-A) in hemostasis inflammatory thrombosis and atherosclerosis" and "Should we forget about platelets and move to leukocytes?"

The synthesis and biological evaluation of new diadenosine polyphosphate analogues on blood platelet aggregation are reported. The most active are compounds with a sulfur atom replacing one or both non-bridging oxygens at phosphorus bound to adenosyl residues and hydroxymethyl groups of bis(hydroxymethyl)phosphinic acid. [Copyright &y& Elsevier]

NiTi shape memory alloys are increasingly being used as bone and cardiac implants. The oxide layer of nanometric thickness spontaneously formed on their surface does not sufficiently protect from nickel transition into surrounding tissues, and its presence, even in a small amount, can be harmful to the human organism. In order to limit this disadvantageous phenomenon, there are several surface engineering techniques used, including oxidation methods. Due to the usually complex shapes of implants, one of the most prospective methods is low-temperature plasma oxidation. This article presents the role of cathode sputtering in the formation of a titanium dioxide surface layer, specifically rutile. The surface of the NiTi shape memory alloy was modified using low-temperature glow discharge plasma oxidation processes, which were carried out in two variants: oxidation using an argon + oxygen (80% vol.) reactive atmosphere and the less chemically active argon + air (80% vol.), but with a preliminary cathode sputtering process in the Ar + N2 (1:1) plasma. This paper presents the structure (STEM), chemical composition (EDS, SIMS), surface topography (optical profilometer, Atomic Force Microscopy—AFM) and antibacterial properties of nanocrystalline TiO2 diffusive surface layers. It is shown that prior cathodic sputtering in argon-nitrogen plasma almost doubled the thickness of the produced nitrogen-doped titanium dioxide layers despite using air instead of oxygen. The (TiOxNy)2 diffusive surface layer showed a high level of resistance to E. coli colonization in comparison with NiTi, which indicates the possibility of using this surface layer in the modification of NiTi implants' properties. [ABSTRACT FROM AUTHOR]

Comparative analysis of the protein profile of blood platelets isolated from nondialyzed and hemodialyzed uremic patients and healthy controls has been performed. These preliminary results indicate markedly changed expression of several proteins in blood platelets from both groups of patients compared with controls, with dramatic changes in hemodialyzed patients in the over-expression of low molecular peptides with a very wide range of pI values. [ABSTRACT FROM AUTHOR]

Biological acceptance is one of the most important aspects of a biomaterial and forms the basis for its clinical use. The aim of this study was a comprehensive biological evaluation (cytotoxicity test, bacterial colonization test, blood platelets adhesion test and transcriptome and proteome analysis of Saos-2 cells after contact with surface of the biomaterial) of biomaterials used in spinal and orthopedic surgery, namely, Ti6Al4V ELI (Extra Low Interstitials), its modified version obtained as a result of melting by electron beam technology (Ti6Al4V ELI-EBT), polyether ether ketone (PEEK) and polished medical steel American Iron and Steel Institute (AISI) 316L (the reference material). Biological tests were carried out using the osteoblasts-like cells (Saos-2, ATCC HTB-85) and bacteria Escherichia coli (DH5α). Results showed lack of cytotoxicity of all materials and the surfaces of both Ti6Al4V ELI and PEEK exhibit a significantly higher resistance to colonization with E. coli cells, while the more porous surface of the same titanium alloy produced by electron beam technology (EBT) is more susceptible to microbial colonization than the control surface of polished medical steel. None of the tested materials showed high toxicity in relation to E. coli cells. Susceptibility to platelet adhesion was very high for polished medical steel AISI 316L, whilst much lower for the other biomaterials and can be ranked from the lowest to the highest as follows: PEEK < Ti6Al4V ELI < Ti6Al4V ELI-EBT. The number of expressed genes in Saos-2 cells exposed to contact with the examined biomaterials reached 9463 genes in total (ranging from 8455 genes expressed in cells exposed to ELI to 9160 genes in cells exposed to PEEK). Whereas the number of differentially expressed proteins detected on two-dimensional electrophoresis gels in Saos-2 cells after contact with the examined biomaterials was 141 for PEEK, 223 for Ti6Al4V ELI and 133 for Ti6Al4V ELI-EBT. Finally, 14 proteins with altered expression were identified by mass spectrometry. In conclusion, none of the tested biomaterials showed unsatisfactory levels of cytotoxicity. The gene and protein expression analysis, that represents a completely new approach towards characterization of these biomaterials, showed that the polymer PEEK causes much more intense changes in gene and protein expression and thus influences cell metabolism. [ABSTRACT FROM AUTHOR]