Effect of a single immersion in cold water below 4 °C on haemorheological properties of blood in healthy men.

Cold water immersion (CWI) involves rapid cooling of the body, which, in healthy individuals, triggers a defence response to an extreme stimulus, to which the body reacts with stress. The aim of the study was to determine the effect of CWI on hemorheological blood indicators. The study group consisted of 13 young males. Blood samples were collected before and after CWI. The assessed parameters included the complete blood count, fibrinogen, hs-C-reactive protein (CRP), proteinogram, and blood rheology factors, such as erythrocyte elongation index (EI), half-time of total aggregation, and aggregation index. Additionally, the effect of reduced temperature on primary human vascular endothelium was investigated in vitro. CWI resulted in the decrease of body temperature to 31.55 ± 2.87 °C. After CWI, neutrophil count and mean corpuscular volume (MCV) were significantly increased in the study group, while lymphocyte count was significantly decreased. Significantly higher levels of total blood protein and albumin concentration were detected after the immersion. Among hemorheological characteristics, erythrocyte EIs at shear stress values ranging from 2.19 to 60.30 Pa were significantly lower after CWI. No significant changes in other rheological, morphological or biochemical parameters were observed. In vitro, human umbilical vein endothelial cells responded to 3 h of temperature decrease to 25 °C with unchanged viability, but increased recruitment of THP-1 monocytic cells and changes in cell morphology were observed. This was the first study to evaluate the effect of single CWI on rheological properties of blood in healthy young men. The results indicate that a single CWI may increase blood protein concentrations and worsen erythrocyte deformability parameters.

A Comparative Study of Women With Substance Use Disorder With and Without Post-traumatic Stress Disorder: Complex Interaction Between Childhood Trauma, Empathy, Personality Traits, and Substance Use Severity.

Objective: Substance use disorders (SUDs) and post-traumatic stress disorder (PTSD) significantly coexist; thus, identifying potential related factors of both illnesses is important for advancing the prevention strategies. The goals of this study were to compare women with SUD with and without PTSD in terms of potentially related variables and to investigate if those factors are associated with SUD severity. Methods: Ninety-five participants (women) with SUD who had been admitted to the outpatient treatment clinic in Turkey were assessed with borderline personality questionnaire, childhood trauma questionnaire, addiction profile index, Levenson self-report psychopathy questionnaire, and basic empathy scale. Results: Participants with PTSD had significantly higher severity of addiction, childhood trauma, borderline personality traits, secondary psychopathy scores, and affective empathy scores than participants without PTSD (P = .013, P = .012, P = .008, P = .031, P = .040). Conclusion: Our study suggested that comorbid PTSD and SUD resulted in more complicated presentation. It seems to be crucial for practitioners to screen PTSD symptoms in women with severe SUD and complicated clinical presentations.

Endothelialization of Whey Protein Isolate-Based Scaffolds for Tissue Regeneration.

BACKGROUND: Whey protein isolate (WPI) is a by-product from the dairy industry, whose main component is ß-lactoglobulin. Upon heating, WPI forms a hydrogel which can both support controlled drug delivery and enhance the proliferation and osteogenic differentiation of bone-forming cells. This study makes a novel contribution by evaluating the ability of WPI hydrogels to support the growth of endothelial cells, which are essential for vascularization, which in turn is a pre-requisite for bone regeneration. METHODS: In this study, the proliferation and antioxidant levels in human umbilical vascular endothelial cells (HUVECs) cultured with WPI supplementation were evaluated using real-time cell analysis and flow cytometry. Further, the attachment and growth of HUVECs seeded on WPI-based hydrogels with different concentrations of WPI (15%, 20%, 30%, 40%) were investigated. RESULTS: Supplementation with WPI did not affect the viability or proliferation of HUVECs monitored with real-time cell analysis. At the highest used concentration of WPI (500 µg/mL), a slight induction of ROS production in HUVECs was detected as compared with control samples, but it was not accompanied by alterations in cellular thiol levels. Regarding WPI-based hydrogels, HUVEC adhered and spread on all samples, showing good metabolic activity. Notably, cell number was highest on samples containing 20% and 30% WPI. CONCLUSIONS: The demonstration of the good compatibility of WPI hydrogels with endothelial cells in these experiments is an important step towards promoting the vascularization of hydrogels upon implantation in vivo, which is expected to improve implant outcomes in the future.

Adjusting Degree of Modification and Composition of gelAGE-Based Hydrogels Improves Long-Term Survival and Function of Primary Human Fibroblasts and Endothelial Cells in 3D Cultures.

This study aimed to develop a suitable hydrogel-based 3D platform to support long-term culture of primary endothelial cells (ECs) and fibroblasts. Two hydrogel systems based on allyl-modified gelatin (gelAGE), G1MM and G2LH, were cross-linked via thiol-ene click reaction with a four-arm thiolated polyethylene glycol (PEG-4-SH). Compared to G1MM, the G2LH hydrogel was characterized by the lower polymer content and cross-linking density with a softer matrix and homogeneous and open porosity. Cell viability in both hydrogels was comparable, although the G2LH-based platform supported better F-actin organization, cell-cell interactions, and collagen and fibronectin production. In co-cultures, early morphogenesis leading to tubular-like structures was observed within 2 weeks. Migration of fibroblasts out of spheroids embedded in the G2LH hydrogels started after 5 days of incubation. Taken together, the results demonstrated that the G2LH hydrogel fulfilled the demands of both ECs and fibroblasts to enable long-term culture and matrix remodeling.

Polydopamine and gelatin coating for rapid endothelialization of vascular scaffolds.

Rapid endothelialization helps overcome the limitations of small-diameter vascular grafts. To develop biomimetic non-thrombogenic coatings supporting endothelialization, medical-grade polyurethane (PU) nanofibrous mats and tubular scaffolds with a diameter below 6 mm prepared by solution blow spinning were coated with polydopamine (PDA), or PDA and gelatin (PDA/Gel). The scaffolds were characterized by scanning electron microscopy, porosity measurement, tensile testing, wettability, Fourier Transform Infrared spectroscopy, and termogravimetric analysis, followed by the measurement of coating stability on the tubular scaffolds. The effect of coating on scaffold endothelialization and hemocompatibility was evaluated using human umbilical vein endothelial cells (HUVECs) and human platelets, showing low numbers of adhering platelets and significantly higher numbers of HUVECs on PDA- and PDA/Gel-coated mats compared to control samples. Tubular PU scaffolds and commercial ePTFE prostheses coated with PDA or PDA/Gel were colonized with HUVECs using radial magnetic cell seeding. PDA/Gel-coated samples achieved full endothelial coverage within 1-3 days post-endothelialization. Altogether, PDA and PDA/Gel coating significantly enhance the endothelialization on the flat surfaces, tubular small-diameter scaffolds, and commercial vascular prostheses. The presented approach constitutes a fast and efficient method of improving scaffold colonization with endothelial cells, expected to work equally well upon implantation.

A Print-and-Fuse Strategy for Sacrificial Filaments Enables Biomimetically Structured Perfusable Microvascular Networks with Functional Endothelium Inside 3D Hydrogels.

A facile and flexible approach for the integration of biomimetically branched microvasculature within bulk hydrogels is presented. For this, sacrificial scaffolds of thermoresponsive poly(2-cyclopropyl-2-oxazoline) (PcycloPrOx) are created using melt electrowriting (MEW) in an optimized and predictable way and subsequently placed into a customized bioreactor system, which is then filled with a hydrogel precursor solution. The aqueous environment above the lower critical solution temperature (LCST) of PcycloPrOx at 25 °C swells the polymer without dissolving it, resulting in fusion of filaments that are deposited onto each other (print-and-fuse approach). Accordingly, an adequate printing pathway design results in generating physiological-like branchings and channel volumes that approximate Murray's law in the geometrical ratio between parent and daughter vessels. After gel formation, a temperature decrease below the LCST produces interconnected microchannels with distinct inlet and outlet regions. Initial placement of the sacrificial scaffolds in the bioreactors in a pre-defined manner directly yields perfusable structures via leakage-free fluid connections in a reproducible one-step procedure. Using this approach, rapid formation of a tight and biologically functional endothelial layer, as assessed not only through fluorescent dye diffusion, but also by tumor necrosis factor alpha (TNF-α) stimulation, is obtained within three days.

Differential Responses to Bioink-Induced Oxidative Stress in Endothelial Cells and Fibroblasts.

A hydrogel system based on oxidized alginate covalently crosslinked with gelatin (ADA-GEL) has been utilized for different biofabrication approaches to design constructs, in which cell growth, proliferation and migration have been observed. However, cell-bioink interactions are not completely understood and the potential effects of free aldehyde groups on the living cells have not been investigated. In this study, alginate, ADA and ADA-GEL were characterized via FTIR and NMR, and their effect on cell viability was investigated. In the tested cell lines, there was a concentration-dependent effect of oxidation degree on cell viability, with the strongest cytotoxicity observed after 72 h of culture. Subsequently, primary human cells, namely fibroblasts and endothelial cells (ECs) were grown in ADA and ADA-GEL hydrogels to investigate the molecular effects of oxidized material. In ADA, an extremely strong ROS generation resulting in a rapid depletion of cellular thiols was observed in ECs, leading to rapid necrotic cell death. In contrast, less pronounced cytotoxic effects of ADA were noted on human fibroblasts. Human fibroblasts had higher cellular thiol content than primary ECs and entered apoptosis under strong oxidative stress. The presence of gelatin in the hydrogel improved the primary cell survival, likely by reducing the oxidative stress via binding to the CHO groups. Consequently, ADA-GEL was better tolerated than ADA alone. Fibroblasts were able to survive the oxidative stress in ADA-GEL and re-entered the proliferative phase. To the best of our knowledge, this is the first report that shows in detail the relationship between oxidative stress-induced intracellular processes and alginate di-aldehyde-based bioinks.

Biocompatibility of designed MicNo-ZnO particles: Cytotoxicity, genotoxicity and phototoxicity in human skin keratinocyte cells.

Recently, designed platelet shaped micron particles that are composed of nano primary particles, called MicNo (=Micron+naNo) particles, have been developed to exploit the benefits of nano size, while removing the adverse effects of nanoparticles. It has been shown that MicNo-ZnO particles exhibit both micron and nanosized particle characteristics. Although physical and chemical properties of MicNo-ZnO particles have been studied, their biocompatibility has not yet been evaluated. Accordingly, the research objective of this study was to evaluate in vitro cytotoxicity, genotoxicity and phototoxicity behaviors of designed MicNo-ZnO particles over human epidermal keratinocyte (HaCaT) cells. MicNo-ZnO particles exhibit much less cytotoxicity with IC50 concentrations between 40 and 50µg/ml, genotoxicity above 40µg/ml and lower photo genotoxicity under UVA on HaCaT than the ZnO nanoparticles. Although their chemistries are the same, the source of this difference in toxicity values may be attributed to size differences between the particles that are probably due to their ability to penetrate into the cells. In the present study, the expansive and detailed in vitro toxicity tests show that the biocompatibility of MicNo-ZnO particles is much better than that of the ZnO nanoparticles. Consequently, MicNo-ZnO particles can be considered an important active ingredient alternative for sunscreen applications due to their safer characteristics with respect to ZnO nanoparticles.

In vitro evaluation of combination of EGCG and Erlotinib with classical chemotherapeutics on JAR cells.

Gestational Trophoblastic Neoplasia (GTN) is a term used for a group of malignant gynecological tumors including choriocarcinoma. Low-risk neoplasias can be cured using single agents Methotrexate (MTX) and actinomycin-D (ACD), but in certain cases, decreased responsiveness and serious side effects occur. Therefore, researchers have been attempting to find new treatment modalities. One of the most popular way for increasing cancer patient survival rates is supporting treatment with adjuvant molecules or chemosensitizers. For this purpose, we investigated epigallocatechin-3-gallate (EGCG), a green tea cathecin, and Erlotinib, an EGFR tyrosine kinase inhibitor, as single agents and combined with MTX or ACD. In accordance with this, JAR (human placenta choriocarcinoma) cell line was used as an in vitro model and MTT, LDH, caspase-3 activation, RT-PCR, and Western Blot analyses were performed to investigate the effects of the test materials. Our studies demonstrate that combination of Erlotinib and EGCG with MTX and ACD decreases JAR cell proliferation and metastatic HER2 protein synthesis and increases caspase-3 activation compared to ACD or MTX alone. In addition, significant increase was observed in the apoptotic Bax gene, but no notable protein synthesis occurred in the Western Blot analysis, which suggests that combination of Erlotinib and EGCG with classical chemotherapeutics ACD or MTX may lead the JAR cells to apoptosis, but not by a mitochondrial pathway. All the results indicate that the synergetic effect of Erlotinib and EGCG with classical chemotherapeutics may help to increase patient survival rates of choriocarcinoma, but the detailed mechanism needs further investigation.

Analyses of the modulatory effects of antibacterial silver doped calcium phosphate-based ceramic nano-powder on proliferation, survival, and angiogenic capacity of different mammalian cells in vitro.

In this study, the antibacterial, cytotoxic, and angiogenic activities of silver doped calcium phosphate-based inorganic powder (ABT or PAG) were systematically investigated. ABT powders containing varying silver content were fabricated using a wet chemical manufacturing method. Antibacterial efficiencies of the ABT powders were investigated using a standard test with indicator bacteria and yeast. The cytotoxic effects of ABT on three different fibroblast cells and human umbilical vein endothelial cells (HUVECs) were assessed using MTT assay. ABT powder exhibits concentration-related cytotoxicity characteristics. Apoptotic activity, attachment capability, and wound healing effects were examined on fibroblasts. The angiogenic activity of ABT was investigated by tube formation assay in HUVECs; 10 µg ml(-1) and 100 µg ml(-1) concentrations of the highest metal ion content of ABT did not disrupt the tube formation of HUVECs. All these tests showed that ABT does not compromise the survival of the cells and might impose regeneration ability to various cell types. These results indicate that silver doped calcium phosphate-based inorganic powder with an optimal silver content has good potential for developing new biomaterials for implant applications.

Studies on the cytotoxic, apoptotic and antitumoral effects of Au(III) and Pt(II) complexes of 1, 10-phenanthroline on V79 379A and A549 cell lines.

In the present study, Au(III) and Pt(II) complexes of 1, 10-phenanthroline (phen) were synthesized and used as the test compounds. The structure elucidation of the synthesized compounds was performed by IR, (1)H-NMR and MASS spectroscopic data and the results of elemental analyses. The cytotoxic and apoptotic effects of test compounds were elucidated on V79 379A (Chinese hamster lung fibroblast like) and A549 (human lung carcinoma epithelial like) cell lines. Cytotoxicity was measured with MTT assay and antitumoral effect was determined by colony forming ability methods. In addition, nuclear fragmentation and activation of apoptotic enzyme (caspase-3) and DAPI staining were used to detect the apoptotic effect of the compounds. All the test compounds induced time and concentration-dependent cytotoxic and antitumoral effects. Significant increases in the levels of apoptosis were observed with increasing exposure concentration.