scholarly journals Nitric-Acid Oxidized Single-Walled Carbon Nanohorns as a Potential Material for Bio-Applications—Toxicity and Hemocompatibility Studies

Materials ◽  
2021 ◽  
Vol 14 (6) ◽  
pp. 1419
Author(s):  
Wojciech Zieba ◽  
Joanna Czarnecka ◽  
Tomasz Rusak ◽  
Monika Zieba ◽  
Artur P. Terzyk
Keyword(s):  
Membrane Integrity ◽  
Neutral Red ◽  
Dermal Fibroblasts ◽  
Cell Membrane Integrity ◽  
Carbon Nanohorns ◽  
Single Walled Carbon ◽  
Transmission Electron ◽  
Diphenyltetrazolium Bromide ◽  
Application Potential

The results of in vitro studies of single-walled carbon nanohorn (SWCNH) oxidized materials’ cytotoxicity obtained by the cell membrane integrity (Neutral Red Uptake (NRU)) and metabolic activity (by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)) on A549 and human dermal fibroblasts (HDF) cell lines are presented. We also present hemocompatibility studies on human and porcine blood, and an erythrocyte concentrate to prove that the obtained samples will not interfere with blood components. Characterization of the materials is supplemented by ζ-potential measurements, Transmission Electron Microscope (TEM) imaging, and thermogravimetric studies (TG). The presented results show the correlation between the specific surface area of materials and the platelet aggregation, when the ID/IG ratio determined from Raman spectra correlates with hemoglobin release from the erythrocytes (in whole blood testing). A plausible mechanism explaining the observed correlations is given. The cytotoxicity and hemocompatibility studies prove that the studied materials are acceptable for use in biomedical applications, especially a sample SWCNH-ox-1.5 with the best application potential.

scholarly journals Essential Oil Microcapsules Immobilized on Textiles and Certain Induced Effects

Materials ◽  
2019 ◽  
Vol 12 (12) ◽  
pp. 2029 ◽  
Author(s):  
Miruna S. Stan ◽  
Laura Chirila ◽  
Alina Popescu ◽  
Denisa M. Radulescu ◽  
Diana E. Radulescu ◽  
...  
Keyword(s):  
Membrane Integrity ◽  
Dermal Fibroblasts ◽  
Cell Membrane Integrity ◽  
Textile Materials ◽  
Skin Cells ◽  
In Vitro Biocompatibility ◽  
Short Term Exposure ◽  
Before And After

In order to obtain textile materials with potential utility in the development of cosmetic textiles, this study examined the deposition by padding of rose and sage microcapsules on woven textile structures, with different fiber compositions (100% cotton and 50% cotton/50% polyester). Cationization of the textile materials was performed to enhance the degree of uptake the pf the microcapsules on the fabrics’ surface. A commercially acrylate-based binder was used to fix the microcapsules to the textile substrate and to improve the durability against external factors. The finished textile materials were characterized in terms of their physical-mechanical characteristics. The distribution of microcapsules on the fabrics surface before and after five washing cycles and 1000 abrasion cycles was investigated by scanning electron microscopy. The biocompatibility in terms of cell viability, cell membrane integrity and inflammation status of the functionalized fabrics was evaluated on CCD-1070Sk normal human dermal fibroblasts. The cell morphology was evaluated by F-actin staining using fluorescence microscopy and no significant changes were noticed after the incubation in the presence of fabrics compared with control. The in vitro biocompatibility evaluation on human skin cells confirmed the absence of cytotoxicity after the short-term exposure, supporting further in vivo use of these innovative textiles with improved properties.

scholarly journals Effects of plant extracts on antioxidant status and oxidant-induced stress in Caco-2 cells

2007 ◽  
Vol 97 (2) ◽  
pp. 321-328 ◽  
Author(s):  
S. Aisling Aherne ◽  
Joseph P. Kerry ◽  
Nora M. O'Brien
Keyword(s):  
Dna Damage ◽  
Plant Extract ◽  
Plant Extracts ◽  
Antioxidant Status ◽  
Cell Injury ◽  
Membrane Integrity ◽  
Neutral Red ◽  
Dna Integrity ◽  
Cell Membrane Integrity ◽  
Wide Range

Experimental evidence suggests that most herbs and spices possess a wide range of biological and pharmacological activities that may protect tissues against O2-induced damage. The objectives of the present study were: first, to determine the effects of plant extracts on the viability, membrane integrity, antioxidant status and DNA integrity of Caco-2 cells and second, to investigate the cytoprotective and genoprotective effects of these plant extracts against oxidative stress in Caco-2 cells. The plant extracts examined were rosemary (Rosmarinus officinalis L.), oregano (Origanum vulgare L.), sage (Salvia officinalis L.) and echinacea (Echinacea purpurea L.). Cell membrane integrity was assessed by the lactate dehydrogenase release assay. Viability was determined by the neutral red uptake assay (NRUA) and the concentration of compound that resulted in 50 % cell death (IC50) was calculated. Antioxidant status of the cells was assessed by measuring GSH content, catalase activity and superoxide dismutase activity. To examine their cytoprotective and genoprotective effects, Caco-2 cells were pre-treated with each plant extract for 24 h followed by exposure to H2O2. DNA damage was assessed by the comet assay and cell injury was determined by the NRUA. Rosemary was the most toxic (IC50 123 μg/ml) and echinacea the least toxic (IC50 1421 μg/ml). Sage was the only plant extract to affect the antioxidant status of the cells by increasing GSH content. Sage, oregano and rosemary protected against H2O2-induced DNA damage (olive tail moment and percentage tail DNA), whereas protection against H2O2-induced cytotoxicity was afforded by sage only.

scholarly journals Ivermectin induces autophagy-mediated cell death through the AKT/mTOR signaling pathway in glioma cells

2019 ◽  
Vol 39 (12) ◽  
Author(s):  
Jingjing Liu ◽  
Hongsheng Liang ◽  
Chen Chen ◽  
Xiaoxing Wang ◽  
Faling Qu ◽  
...  
Keyword(s):  
Cell Death ◽  
Molecular Mechanisms ◽  
Anticancer Agent ◽  
Glioma Cells ◽  
Mtor Signaling ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Transmission Electron ◽  
Diphenyltetrazolium Bromide

Abstract Glioma is one of the most common types of primary brain tumors. Ivermectin (IVM), a broad-spectrum antiparasitic drug, has been identified as a novel anticancer agent due to its inhibitory effects on the proliferation of glioma cells in vitro and in vivo. However, the ability of IVM to induce autophagy and its role in glioma cell death remains unclear. The main objective of the present study was to explore autophagy induced by IVM in glioma U251 and C6 cells, and the deep underlying molecular mechanisms. In addition, we examined the effects of autophagy on apoptosis in glioma cells. In the present study, transmission electron microscopy (TEM), immunofluorescence, Western blot and immunohistochemistry were used to evaluate autophagy activated by IVM. Cell viability was measured by 3-(4,5-dimethylthiazol2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and colony formation assay. The apoptosis rate was detected by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Meanwhile, autophagy inhibition was achieved by using chloroquine (CQ). U251-derived xenografts were established for examination of IVM-induced autophagy on glioma in vivo. Taken together, the results of the present study showed that autophagy induced by IVM has a protective effect on cell apoptosis in vitro and in vivo. Mechanistically, IVM induced autophagy through AKT/mTOR signaling and induced energy impairment. Our findings show that IVM is a promising anticancer agent and may be a potential effective treatment for glioma cancers.

Triphenyltin acetate toxicity : a biochemical and ultrastructural study on mouse thymocytes

1996 ◽  
Vol 15 (3) ◽  
pp. 219-225 ◽  
Author(s):  
E. Bollo ◽  
L. Ceppa ◽  
E. Cornaglia ◽  
C. Nebbia ◽  
B. Biolatti ◽  
...  
Keyword(s):  
Colorimetric Assay ◽  
Immunosuppressive Agents ◽  
Primary Cultures ◽  
Lactic Dehydrogenase ◽  
Neutral Red ◽  
Transmission Electron ◽  
Dose Dependent ◽  
Triphenyltin Acetate

1 Triphenyltin acetate (TPTA) has been shown to exert in vivo a selective toxic effect on the immune system. To assess in vitro possible alterations induced by TPTA exposure, primary cultures of mouse thymocytes were incubated up to 24 h with graded amounts (1-12 μM) ofthe organotin. 2 The cytotoxic activity has been evaluated with the MTT colorimetric assay, the neutral red (NR) assay and the lactic dehydrogenase (LDH) cellular release. Cell pellets were fixed with 2.5% glutaraldehyde, resin-embedded and ultrathin sections were observed through transmission electron microscopy. 3 After 2 h of incubation, dose-dependent increases of cytotoxicity were observed in thymocytes submitted to MTT and NR tests (up to 41.43% and 18.9%, respectively), while 22 h later this overt effect on cell viability was noticed merely in cells exposed to 12 μM TPTA. Dose- dependent increases of LDH leakage in the culture medium were observed all throughout the study. 4 Morphological investigations revealed features (chro matin condensation, cell membranes fragmentation and formation of membrane bound apoptotic bodies) sugges tive of apoptosis. 5 This study indicates that TPTA is cytotoxic to mouse thymocytes: morphologically, the rising of apoptosis is likely to be recognized, as previously reported in different in vitro studies with other immunosuppressive agents as dioxin and corticosteroids.

Prodrug Micelles Based on Norbornene-Functional Poly(lactide)s Backbone for Redox-Responsive Release of Paclitaxel

2016 ◽  
Vol 69 (10) ◽  
pp. 1140 ◽  
Author(s):  
Ji Wang ◽  
Jing Yan ◽  
Huicong Zhou ◽  
Haikang Huang ◽  
Xuefei Zhang ◽  
...  
Keyword(s):  
In Vitro Release ◽  
Amphiphilic Copolymer ◽  
Polymer Carriers ◽  
Polymeric Scaffold ◽  
Transmission Electron ◽  
Release Studies ◽  
Diphenyltetrazolium Bromide ◽  
Redox Responsive ◽  
Vitro Release

Norbornene-functional poly(lactide)s backbone-based amphiphilic copolymer, P(LA-g-mOEG)-b-P(LA-SS-COOH), was synthesized as the polymeric scaffold and paclitaxel (PTX) was directly conjugated to the carboxyl groups of the amphiphilic copolymer to obtain redox-responsive P(LA-g-mOEG)-b-P(LA-SS-PTX) prodrugs. The dynamic light scattering and transmission electron microscopy analyses showed that P(LA-g-mOEG)-b-P(LA-SS-PTX) self-assembled into prodrug micelles with a diameter of 60–70 nm and a low polydispersity in aqueous solution. Remarkably, in vitro release studies revealed that 80 % of PTX was released in 72 h under a reductive environment, whereas only 23 % of PTX was released in 72 h under non-reductive conditions. In addition, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays showed that P(LA-g-mOEG)-b-P(LA-SS-PTX) prodrug micelles retained high anti-tumour activity while polymer carriers were non-toxic up to a tested concentration of 1.0 mg mL–1. These redox-responsive prodrug micelles have tremendous potential for anti-tumour drug delivery.

scholarly journals Contribution of the band 3-ankyrin interaction to erythrocyte membrane mechanical stability

Blood ◽  
1991 ◽  
Vol 77 (7) ◽  
pp. 1581-1586 ◽  
Author(s):  
PS Low ◽  
BM Willardson ◽  
N Mohandas ◽  
M Rossi ◽  
S Shohet
Keyword(s):  
Blood Cell ◽  
Red Blood Cell ◽  
Mechanical Stability ◽  
Membrane Integrity ◽  
Membrane Skeleton ◽  
Band 3 ◽  
Cell Membrane Integrity ◽  
The Face ◽  
Cell Stability

Abstract In an effort to evaluate the role of the band 3-ankyrin linkage in maintenance of red blood cell membrane integrity, solution conditions were sought that would selectively dissociate the band 3-ankyrin linkage, leaving other membrane skeletal interactions intact. For this purpose erythrocytes were equilibrated overnight in nutrient-containing buffers at a range of elevated pHs and then examined for changes in mechanical stability and membrane skeletal composition. Band 3 was found to be released from interaction with the membrane skeleton over a pH range (8.4 to 9.5) that was observed to dissociate the band 3- ankyrin interaction in vitro. In contrast, all other membrane skeletal associations appeared to remain intact up to pH 9.3, after which they were also seen to dissociate. Whereas hemolysis of mechanically unstressed cells did not begin until approximately pH 9.3, where the membrane skeletons began to disintegrate, enhanced fragmentation of shear stressed membranes was seen to begin near pH 8, where band 3 dissociation was first observed. Furthermore, the shear-induced fragmentation rate was found to reach a maximum at pH 9.4, ie, where band 3 dissociation was essentially complete. Based on these correlations, we hypothesize that the band 3-ankyrin linkage of the membrane skeleton to the lipid bilayer is essential for red blood cell stability in the face of mechanical distortion but not for cellular integrity in the absence of mechanical stress.

scholarly journals Two Polyketides Produced by Endophytic Penicillium citrinum DBR-9 From Medicinal Plant Stephania kwangsiensis and Their Antifungal Activity Against Plant Pathogenic Fungi

2019 ◽  
Vol 14 (5) ◽  
pp. 1934578X1984679 ◽  
Author(s):  
Haiyu Luo ◽  
Zhen Qing ◽  
Yecheng Deng ◽  
Zhiyong Deng ◽  
Xia’an Tang ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
Membrane Integrity ◽  
Pathogenic Fungi ◽  
Significant Inhibition ◽  
Penicillium Citrinum ◽  
Plant Pathogenic Fungi ◽  
Fungal Cell ◽  
Cell Membrane Integrity ◽  
Chinese Medicinal Plant

Endophytic fungi, especially those found in medicinal plants, are widely studied as producers of secondary metabolites of biotechnological interest. In this study, on the basis of an activity-directed isolation method and spectroscopic analysis, two active polyketides, citrinin (1) and emodin (2), were isolated and identified from the fermentation of the endophytic fungus Penicillium citrinum DBR-9. This fungus was isolated from the root tubers of the traditional Chinese medicinal plant Stephania kwangsiensis. In vitro antifungal assay showed that the two polyketides displayed significant inhibition on hypha growth of tested plant pathogenic fungi with IC50 values ranging from 3.1 to 123.1 μg/mL and 3.0 to 141.0 μg/mL, respectively. In addition, the mechanism of the effects of emodin (2) on the pathogen revealed it could affect the colony morphology, destroy cell membrane integrity, and influence the protein synthesis of the tested fungal cell. This work is the first report of two polyketides-producing endophytic P. citrinum DBR-9 from the medicinal plant S. kwangsiensis. Our results present new opportunities to deeply understand the potential of these two polyketides as natural antifungal agents to control phytopathogens in agriculture.

scholarly journals Osteoblasts Growth Behaviour on Bio-Based Calcium Carbonate Aragonite Nanocrystal

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Abdullahi Shafiu Kamba ◽  
Zuki Abu Bakar Zakaria
Keyword(s):  
Calcium Carbonate ◽  
Fluorescent Microscopy ◽  
Membrane Integrity ◽  
Biological Response ◽  
Cellular Interactions ◽  
Cell Membrane Integrity ◽  
Native Bone ◽  
Composition Structure ◽  
Stimulate Osteoblast Differentiation

Calcium carbonate (CaCO3) nanocrystals derived from cockle shells emerge to present a good concert in bone tissue engineering because of their potential to mimic the composition, structure, and properties of native bone. The aim of this study was to evaluate the biological response of CaCO3nanocrystals on hFOB 1.19 and MC3T3 E-1 osteoblast cellsin vitro. Cell viability and proliferation were assessed by MTT and BrdU assays, and LDH was measured to determine the effect of CaCO3nanocrystals on cell membrane integrity. Cellular morphology was examined by SEM and fluorescence microscopy. The results showed that CaCO3nanocrystals had no toxic effects to some extent. Cell proliferation, alkaline phosphatase activity, and protein synthesis were enhanced by the nanocrystals when compared to the control. Cellular interactions were improved, as indicated by SEM and fluorescent microscopy. The production of VEGF and TGF-1 was also affected by the CaCO3nanocrystals. Therefore, bio-based CaCO3nanocrystals were shown to stimulate osteoblast differentiation and improve the osteointegration process.

Soluble CD14 Protects Human Lymphocytes from Apoptosis

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2566-2566
Author(s):  
Elizabeth Naparstek ◽  
Benjamin Sredni ◽  
Eti Zigman ◽  
Gali Senyor ◽  
Boris Tartakovsky
Keyword(s):  
Human Peripheral Blood ◽  
Protective Efficacy ◽  
Human Lymphocytes ◽  
Membrane Integrity ◽  
Peripheral Blood Lymphocytes ◽  
Apoptotic Cells ◽  
Soluble Cd14 ◽  
Apoptotic Pathway ◽  
Cell Membrane Integrity

Abstract CD14, a 56 Kd glycoprotein, typically present on myeloid cells, has been traditionally associated with innate immunity and pattern recognition. Recently its membrane bound form has been shown to be involved in apoptosis, as a tethering receptor for apoptotic cells on the surface of phagocytes-in this case with the purpose of removing apoptotic cells, and also as a surface molecule involved in protection from apoptosis of monocytes, neutrophils and recently on enterocytes, challenged with LPS. Our aim was to evaluate the possible involvement of the soluble CD14 in the apoptotic pathway of human lymphocytes. Methods: Freshly obtained human peripheral blood lymphocytes were cultured in vitro with gliotoxin, an apoptotic inducer. Human recombinant CD14 was added to the culture at physiological concentrations (10μg/ml-0.5 μg/ml) and apoptosis was assessed by cell membrane integrity using 7AAD, mitochondrial membrane potential by DiOC6(3) and cytoplasm shrinkage by cell size scatter analysis. Results: Using DiOC6(3) we were able to show that human lymphocytes cultured in the presence of gliotoxin contained 63.8%±21 apoptotic cells, as opposed to 12.2%±11.5 in control cultures. Addition of recombinant human CD14 at a concentration of 10 mg/ml neutralized the apoptotic effect of gliotoxin back to 20.2%±10 (p<0.003). This inhibitory effect was blocked by CD14-specific monoclonal antibodies, but not by control antibodies. We then identified and synthesized the fragment within the CD14 molecule that was responsible for this apoptosis protective effect, and demonstrated its comparable protective efficacy in vitro as shown in figure 1. The figure clearly reveals that this specific peptide, as opposed to the scrambled peptide, protected the lymphocytes form apoptosis, similarly to the full CD14 protein. Same results were obtained using 7AAD and cytoplasm shrinkage. Conclusion: Our data thus suggest that circulating CD14 may play an important role in the prevention of apoptosis of lymphocytes and perhaps of other cells. Figure Figure

Assessment of changes in sperm cell membrane integrity occurring during the storage of semen from genetically different males using two diagnostic methods

2014 ◽  
Vol 94 (4) ◽  
pp. 601-606 ◽  
Author(s):  
Anna Wysokińska ◽  
Stanislaw Kondracki
Keyword(s):  
Cell Membrane ◽  
Sperm Cell ◽  
Cell Membranes ◽  
Intact Cell ◽  
Membrane Integrity ◽  
Diagnostic Methods ◽  
Cell Membrane Integrity ◽  
Staining Methods ◽  
Breed Crosses

Wysokińska, A. and Kondracki, S. 2014. Assessment of changes in sperm cell membrane integrity occurring during the storage of semen from genetically different males using two diagnostic methods. Can. J. Anim. Sci. 94: 601–606. The present study was carried out to assess changes in sperm cell membrane integrity occurring during the storage of semen collected from genetically different domestic male pigs. The study was aimed at assessing differences in the course of changes in the integrity of cell membranes in spermatozoa produced by males with different degrees of genetic diversity (pure-bred males, two-breed hybrids and multi-breed crosses) and testing the usefulness of two methods of sperm cell membrane integrity evaluation, based on material collected from genetically different males. The experiments were conducted on 56 ejaculates collected from 28 domestic male pigs. The examination of sperm cell membrane integrity was performed three times for each ejaculate, i.e., after 1 h, after 24 h and after 48 h from collection. The preparations for analysing cell membrane integrity were made using two methods: the SYBR 14/PI method and the eosin–nigrosin method. It was found that both SYBR 14/PI and eosin–nigrosin staining methods make it possible to successfully assess the integrity of the plasma membrane of domestic pig sperm cells under in vitro conditions. Hybrid pig spermatozoa, especially those from multi-breed crosses, better retain the integrity of their plasmalemmas than the spermatozoa of pure-bred boars. The ejaculates of Hypor cross-breed boars assessed after 1, 24 and 48 h of storage contain more spermatozoa with intact cell membranes than the ejaculates of pure-bred Duroc and Pietrain boars. The ejaculates of Hypor boars also show fewer decaying spermatozoa than those produced by pure-bred boars.

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