Assessment of the Toxicity of Biocompatible Materials Supporting Bone Regeneration: Impact of the Type of Assay and Used Controls

Assessing the toxicity of new biomaterials dedicated to bone regeneration can be difficult. Many reports focus only on a single toxicity parameter, which may be insufficient for a detailed evaluation of the new material. Moreover, published data frequently do not include control cells exposed to the environment without composite or its extract. Here we present the results of two assays used in the toxicological assessment of materials’ extracts (the integrity of the cellular membrane and the mitochondrial activity/proliferation), and the influence of different types of controls used on the obtained results. Results obtained in the cellular membrane integrity assay showed a lack of toxic effects of all tested extracts, and no statistical differences between them were present. Control cells, cells incubated with chitosan extract or chitosan-bioglass extract were used as a reference in proliferation calculations to highlight the impact of controls used on the result of the experiment. The use of different baseline controls caused variability between obtained proliferation results, and influenced the outcome of statistical analysis. Our findings confirm the thesis that the type of control used in an experiment can change the final results, and it may affect the toxicological assessment of biomaterial.

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Synthesis, Spectroscopy, Light Stability, Single-Crystal Analysis, and In Vitro Cytotoxic Activity on HepG2 Liver Cancer of Two Novel Silver(I) Complexes of Miconazole

International Journal of Molecular Sciences ◽

10.3390/ijms21103629 ◽

2020 ◽

Vol 21 (10) ◽

pp. 3629

Author(s):

Karolina Stryjska ◽

Lidia Radko ◽

Lilianna Chęcińska ◽

Joachim Kusz ◽

Andrzej Posyniak ◽

...

Keyword(s):

Cancer Cells ◽

Human Cancer ◽

Membrane Damage ◽

Membrane Integrity ◽

Free Ligand ◽

Cellular Membrane ◽

Biologically Active ◽

Total Protein Content ◽

Mitochondrial Activity ◽

Cytotoxic Activities

Two novel silver(I) complexes of the biologically active ligand miconazole in the form of Ag(MCZ)2X (MCZ = 1-[2-(2,4-dichlorobenzyloxy)-2-(2,4-dichlorophenyl)ethyl]-1H-imidazole]; X = NO3− (1), ClO4− (2)) were synthesized and fully characterized. The complexes were obtained by reactions of Ag(I) salts with miconazole (MCZ). Silver(I) complexes were characterized by elemental analysis, 1H-NMR and infrared (IR) spectroscopy, electrospray ionization (ESI)-MS spectrometry, and X-ray-crystallography. This work also presents a cytotoxicity study of the silver(I) complexes of miconazole and appropriate silver(I) salts using Balb/c 3T3 and HepG2 cell lines. The cytotoxicity of the compounds was assessed based on four biochemical endpoints: lysosomal activity (neutral red uptake (NRU) assay), mitochondrial activity (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay), total protein content (TPC assay), and cellular membrane integrity (lactate dehydrogenase (LDH) assay). The cancer HepG2 cells were more sensitive to the complexes tested, and the most affected endpoint was cellular membrane damage compared to Balb/c 3T3 fibroblasts. Moreover, study complexes inhibited the growth of cancer cells at submicromolecular concentrations (0.26–0.47 μM) lower than that required for the anticancer agent, cisplatin, in MTT, NRU, and TPC assays. Both complexes were characterized by higher toxicity to human cancer cells (HepG2) than silver(I) salts and the free ligand. Combination of Ag(I) salts with miconazole is associated with the marked improvement of cytotoxic activities that can be considered as the significant point in the construction of a new generation of antineoplastic agents.

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Influence of fluoroquinolones on viability of Balb/c 3T3 and HepG2 cells

Bulletin of the Veterinary Institute in Pulawy ◽

10.2478/bvip-2013-0102 ◽

2013 ◽

Vol 57 (4) ◽

pp. 599-606 ◽

Cited By ~ 5

Author(s):

Lidia Radko ◽

Maria Minta ◽

Sylwia Stypuła-Trębas

Keyword(s):

Cell Lines ◽

Membrane Integrity ◽

Cellular Membrane ◽

Neutral Red ◽

Total Protein Content ◽

Mitochondrial Activity ◽

Response Curves ◽

Lactate Dehydrogenase Release ◽

Hepg2 Cell Lines ◽

Hepatoma Hepg2

Abstract The cytotoxic potential of fluoroquinolones (enrofloxacin, ciprofloxacin, difloxacin, sarafloxacin, danofloxacin, norfloxacin and marbofloxacin) was investigated using mouse fibroblasts Balb/c 3T3 and human hepatoma HepG2 cell lines. The cells were exposed for 24, 48, and 72 h to drugs at eight concentrations ranged from 0.78 to 100 μg/mL. Four independent cytotoxicity assays were applied, in which various endpoints were assessed: mitochondrial activity - MTT reduction, lysosomal activity - neutral red uptake, total protein content, and cellular membrane integrity - lactate dehydrogenase release. Mean effective cytotoxic concentrations (EC50) calculated at different time points from concentration-response curves ranged from 10 to 100 μg/mL. The most affected endpoint in both cell lines was mitochondrial activity. The EC50-MTT-72 h <10 μg/mL was found for difloxacin, marbofloxacin (fibroblasts), sarafloxacin, and norfloxacin (HepG2). The data shows that cytotoxicity of the fluoroquinolones appears after longer exposure of both cell cultures to these compounds.

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Cytotoxicity of Mycotoxins Frequently Present in Aquafeeds to the Fish Cell Line RTGill-W1

Toxins ◽

10.3390/toxins13080581 ◽

2021 ◽

Vol 13 (8) ◽

pp. 581

Author(s):

Elena Bernal-Algaba ◽

Marta Pulgarín-Alfaro ◽

María Luisa Fernández-Cruz

Keyword(s):

Plasma Membrane ◽

Cell Line ◽

Membrane Integrity ◽

Acute Effect ◽

Economic Problem ◽

Fish Cell Line ◽

Mitochondrial Activity ◽

Fish Cell ◽

Aquaculture Industry ◽

The Impact

In the last decades, the aquaculture industry has introduced plant-based ingredients as a source of protein in aquafeeds. This has led to mycotoxin contaminations, representing an ecological, health and economic problem. The aim of this study was to determine in the RTgill-W1 fish cell line the toxicity of fifteen mycotoxins of common occurrence in aquafeeds. To identify the most sensitive endpoint of toxicity, the triple assay was used. It consisted of three assays: alamarBlue, Neutral Red Uptake and CFDA-AM, which revealed the mitochondrial activity, the lysosomal integrity and the plasma membrane integrity, respectively. Most of the assayed mycotoxins were toxic predominantly at lysosomal level (enniatins, beauvericin, zearalenone, ochratoxin A, deoxynivalenol (DON) and its acetylated metabolites 15-O-acetyl-DON and 3-acetyl-DON). Aflatoxins B1 and B2 exerted the greatest effects at mitochondrial level, while fumonisins B1 and B2 and nivalenol were not toxic up to 100 µg/mL. In general, low toxicity was observed at plasma membrane level. The vast majority of the mycotoxins assayed exerted a pronounced acute effect in the fish RTgill-W1 cell line, emphasizing the need for further studies to ascertain the impact of mycotoxin contamination of fish feeds in the aquaculture industry and to establish safe limits in aquafeeds.

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Quota Women Are Threatening to Men

Swiss Journal of Psychology ◽

10.1024/1421-0185/a000195 ◽

2017 ◽

Vol 76 (3) ◽

pp. 107-116 ◽

Cited By ~ 4

Author(s):

Klea Faniko ◽

Till Burckhardt ◽

Oriane Sarrasin ◽

Fabio Lorenzi-Cioldi ◽

Siri Øyslebø Sørensen ◽

...

Keyword(s):

Decision Making ◽

Organizational Performance ◽

Gender Stereotypes ◽

Perceived Threat ◽

Preferential Treatment ◽

Public Sector Employees ◽

Different Types ◽

Low Levels ◽

Women In Decision Making ◽

The Impact

Abstract. Two studies carried out among Albanian public-sector employees examined the impact of different types of affirmative action policies (AAPs) on (counter)stereotypical perceptions of women in decision-making positions. Study 1 (N = 178) revealed that participants – especially women – perceived women in decision-making positions as more masculine (i.e., agentic) than feminine (i.e., communal). Study 2 (N = 239) showed that different types of AA had different effects on the attribution of gender stereotypes to AAP beneficiaries: Women benefiting from a quota policy were perceived as being more communal than agentic, while those benefiting from weak preferential treatment were perceived as being more agentic than communal. Furthermore, we examined how the belief that AAPs threaten men’s access to decision-making positions influenced the attribution of these traits to AAP beneficiaries. The results showed that men who reported high levels of perceived threat, as compared to men who reported low levels of perceived threat, attributed more communal than agentic traits to the beneficiaries of quotas. These findings suggest that AAPs may have created a backlash against its beneficiaries by emphasizing gender-stereotypical or counterstereotypical traits. Thus, the framing of AAPs, for instance, as a matter of enhancing organizational performance, in the process of policy making and implementation, may be a crucial tool to countering potential backlash.

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Situational Breakdowns

10.1093/oso/9780190922061.001.0001 ◽

2019 ◽

Cited By ~ 8

Author(s):

Anne Nassauer

Keyword(s):

United States ◽

Large Scale ◽

Contextual Factors ◽

The United States ◽

Social Outcomes ◽

Dynamic Processes ◽

Collective Decisions ◽

Different Types ◽

The Impact

This book provides an account of how and why routine interactions break down and how such situational breakdowns lead to protest violence and other types of surprising social outcomes. It takes a close-up look at the dynamic processes of how situations unfold and compares their role to that of motivations, strategies, and other contextual factors. The book discusses factors that can draw us into violent situations and describes how and why we make uncommon individual and collective decisions. Covering different types of surprise outcomes from protest marches and uprisings turning violent to robbers failing to rob a store at gunpoint, it shows how unfolding situations can override our motivations and strategies and how emotions and culture, as well as rational thinking, still play a part in these events. The first chapters study protest violence in Germany and the United States from 1960 until 2010, taking a detailed look at what happens between the start of a protest and the eruption of violence or its peaceful conclusion. They compare the impact of such dynamics to the role of police strategies and culture, protesters’ claims and violent motivations, the black bloc and agents provocateurs. The analysis shows how violence is triggered, what determines its intensity, and which measures can avoid its outbreak. The book explores whether we find similar situational patterns leading to surprising outcomes in other types of small- and large-scale events: uprisings turning violent, such as Ferguson in 2014 and Baltimore in 2015, and failed armed store robberies.

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ATP13A2 Regulates Cellular α-Synuclein Multimerization, Membrane Association, and Externalization

International Journal of Molecular Sciences ◽

10.3390/ijms22052689 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2689

Author(s):

Jianmin Si ◽

Chris Van den Haute ◽

Evy Lobbestael ◽

Shaun Martin ◽

Sarah van Veen ◽

...

Keyword(s):

Membrane Integrity ◽

Ubiquitin Proteasome System ◽

Regulatory Function ◽

Membrane Association ◽

Loss Of Function ◽

Atypical Form ◽

Polyamine Transport ◽

Independent Functions ◽

Ubiquitin Proteasome ◽

The Impact

ATP13A2, a late endo-/lysosomal polyamine transporter, is implicated in a variety of neurodegenerative diseases, including Parkinson’s disease and Kufor–Rakeb syndrome, an early-onset atypical form of parkinsonism. Loss-of-function mutations in ATP13A2 result in lysosomal deficiency as a consequence of impaired lysosomal export of the polyamines spermine/spermidine. Furthermore, accumulating evidence suggests the involvement of ATP13A2 in regulating the fate of α-synuclein, such as cytoplasmic accumulation and external release. However, no consensus has yet been reached on the mechanisms underlying these effects. Here, we aimed to gain more insight into how ATP13A2 is linked to α-synuclein biology in cell models with modified ATP13A2 activity. We found that loss of ATP13A2 impairs lysosomal membrane integrity and induces α-synuclein multimerization at the membrane, which is enhanced in conditions of oxidative stress or exposure to spermine. In contrast, overexpression of ATP13A2 wildtype (WT) had a protective effect on α-synuclein multimerization, which corresponded with reduced αsyn membrane association and stimulation of the ubiquitin-proteasome system. We also found that ATP13A2 promoted the secretion of α-synuclein through nanovesicles. Interestingly, the catalytically inactive ATP13A2 D508N mutant also affected polyubiquitination and externalization of α-synuclein multimers, suggesting a regulatory function independent of the ATPase and transport activity. In conclusion, our study demonstrates the impact of ATP13A2 on α-synuclein multimerization via polyamine transport dependent and independent functions.

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A global analysis of the impact of COVID-19 stay-at-home restrictions on crime

Nature Human Behaviour ◽

10.1038/s41562-021-01139-z ◽

2021 ◽

Author(s):

Amy E. Nivette ◽

Renee Zahnow ◽

Raul Aguilar ◽

Andri Ahven ◽

Shai Amram ◽

...

Keyword(s):

Public Space ◽

Global Analysis ◽

Sudden Change ◽

Interrupted Time Series ◽

Urban Crime ◽

Substantial Variation ◽

Different Types ◽

Time Series Analyses ◽

The Impact ◽

At Home

AbstractThe stay-at-home restrictions to control the spread of COVID-19 led to unparalleled sudden change in daily life, but it is unclear how they affected urban crime globally. We collected data on daily counts of crime in 27 cities across 23 countries in the Americas, Europe, the Middle East and Asia. We conducted interrupted time series analyses to assess the impact of stay-at-home restrictions on different types of crime in each city. Our findings show that the stay-at-home policies were associated with a considerable drop in urban crime, but with substantial variation across cities and types of crime. Meta-regression results showed that more stringent restrictions over movement in public space were predictive of larger declines in crime.

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Comprehensive Effects of Short-Term Rental Platforms Across Hotel Types in U.S. and International Destinations

Cornell Hospitality Quarterly ◽

10.1177/19389655211033527 ◽

2021 ◽

pp. 193896552110335

Author(s):

John W. O’Neill ◽

Jihwan Yeon

Keyword(s):

Service Providers ◽

Comprehensive Analysis ◽

Competitive Market ◽

Substitution Effects ◽

Short Term ◽

Hotel Performance ◽

The World ◽

Different Types ◽

Location Type ◽

The Impact

In recent years, short-term rental platforms in the lodging sector, including Airbnb, VRBO, and HomeAway, have received extensive attention and emerged as potentially alternative suppliers of services traditionally provided by established commercial accommodation providers, that is, hotels. Short-term rentals have dramatically increased the available supply of rooms for visitors to multiple international destinations, potentially siphoning demand away from hotels to short-term rental businesses. In a competitive market, an increase in supply with constant demand would negatively influence incumbent service providers. In this article, we examine the substitution effects of short-term rental supply on hotel performance in different cities around the world. Specifically, we comprehensively investigate the substitution effects of short-term rental supply on hotel performance based on hotel class, location type, and region. Furthermore, we segment the short-term rental supply based on its types of accommodations, that is, shared rooms, private rooms, and entire homes, and both examine and quantify the differential effects of these types of short-term rentals on different types of hotels. This study offers a comprehensive analysis regarding the impact of multiple short-term rental platforms on hotel performance and offers both conceptual and practical insights regarding the nature and extent of the effects that were identified.

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Double, triple or quadruple hits? Exploring the impact of cybercrime on victims in the Netherlands

International Review of Victimology ◽

10.1177/02697580211010692 ◽

2021 ◽

pp. 026975802110106

Author(s):

Raoul Notté ◽

E.R. Leukfeldt ◽

Marijke Malsch

Keyword(s):

Public Opinion ◽

Literature Review ◽

The Netherlands ◽

Public Discourse ◽

Moral Judgments ◽

Victim Blaming ◽

High Prevalence ◽

Different Types ◽

The Impact ◽

Insight Into

This article explores the impact of online crime victimisation. A literature review and 41 interviews – 19 with victims and 22 with experts – were carried out to gain insight into this. The interviews show that most impacts of online offences correspond to the impacts of traditional offline offences. There are also differences with offline crime victimisation. Several forms of impact seem to be specific to victims of online crime: the substantial scale and visibility of victimhood, victimisation that does not stop in time, the interwovenness of online and offline, and victim blaming. Victims suffer from double, triple or even quadruple hits; it is the accumulation of different types of impact, enforced by the limitlessness in time and space, which makes online crime victimisation so extremely invasive. Furthermore, the characteristics of online crime victimisation greatly complicate the fight against and prevention of online crime. Finally, the high prevalence of cybercrime victimisation combined with the severe impact of these crimes seems contradictory with public opinion – and associated moral judgments – on victims. Further research into the dominant public discourse on victimisation and how this affects the functioning of the police and victim support would be valuable.

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Evaluation of a panel of spermatological methods for assessing reprotoxic compounds in multilayer semen plastic bags

Scientific Reports ◽

10.1038/s41598-020-79415-7 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

M. Schulze ◽

F. Schröter ◽

M. Jung ◽

U. Jakop

Keyword(s):

Membrane Integrity ◽

Diglycidyl Ether ◽

Mitochondrial Activity ◽

Prolonged Storage ◽

Prolonged Incubation ◽

Plastic Bags ◽

Plastic Materials ◽

Bisphenol A Diglycidyl Ether ◽

Semen Preservation ◽

Boar Spermatozoa

AbstractThe increase of fertility performance in sows is one of the biggest achievements in pig production over the last 30 years. Nevertheless, pig farms using artificial insemination (AI) repeatedly experienced in recent year’s fertility problems with dramatic consequences due to toxic compounds from plastic semen bags. In particular, bisphenol A diglycidyl-ether (BADGE) present in multilayer plastic bags can leach into the semen and could affect the functionality of the spermatozoa. Former studies could not find any alterations in spermatozoa based on the exposure to BADGE. The aim of the study was to evaluate effects of BADGE on boar spermatozoa using an extended panel of spermatological methods. In spring 2019, a large drop in farrowing rates from 92.6 ± 2.3% to 63.7 ± 11.1% in four sow farms in Croatia was detected. In migration studies, BADGE could be identified as a causal toxic compound and leached into the extended semen in concentration of 0.37 ± 0.05 mg/L. Detailed spermatological studies showed that significant predictors for effects on spermatozoa were different levels of motility and kinematic data after a prolonged storage time, thermo-resistance test (prolonged incubation time), mitochondrial activity, membrane integrity and fluidity. No serious effects were observed for sperm morphology and DNA fragmentation. These results provide new insights into the development of a new quality assurance concept for a detailed spermatological examination during testing of plastic materials for boar semen preservation. It could be shown that boar spermatozoa are an excellent biosensor to detect potential toxicity and fertility-relevant compounds.

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