Real-time monitoring of cadmium toxicity in rabbit kidney cells

The aim of this study was to investigate the toxic effect of the metal salt cadmium chloride dihydrate on the rabbit kidney cell line using the xCELLigence system or real-time cell analyser (RTCA), and to compare this relatively new method with standard biological cytotoxicity assays. This system provides real-time monitoring of cell behaviour and proliferative activity during the whole time of experiment. Moreover, after 24 h exposure of cells to cadmium, colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5-difenyl tetrazolium bromide (MTT) test was used to measure the metabolic activity and cytotoxicity was determined by measurement of lactate dehydrogenase (LDH) leaked from damaged cells. We found that renal cells exposed to lower concentrations (5–10 mg·l-1) of cadmium tend to grow similarly to control cells, however, cell index was significantly different (P < 0.05) after 24 h. With increasing concentration of cadmium (15–50 mg·l-1) significantly lower proliferative (P < 0.05) and metabolic activity (P < 0.05) of cells was observed and cytotoxicity increased simultaneously (P < 0.001). In addition, we found that the real-time monitoring of the cell response was significantly correlated with commonly used biological methods for toxicity measurement, for MTT assay R2 was 0.9448 (P < 0.01) and for LDH assay R2 was 0.9466 (P < 0.01), respectively. The present study is the first report when combination of RTCA, MTT assay and LDH test was used for cadmium nephrotoxicity assessment. In all these methods, the toxic effect of cadmium on rabbit kidney cells increased in a concentration-dependent manner.

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Real-time monitoring of cellular metabolic activity: intracellular oxygen

Nature Methods ◽

10.1038/nmeth.f.396 ◽

2016 ◽

Vol 13 (10) ◽

pp. i-ii

Author(s):

Andrea Krumm ◽

Conn Carey

Keyword(s):

Real Time ◽

Metabolic Activity ◽

Real Time Monitoring ◽

Cellular Metabolic Activity

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Real-time monitoring of bacterial biofilms metabolic activity by a redox-reactive nanosensors array

Journal of Nanobiotechnology ◽

10.1186/s12951-020-00637-y ◽

2020 ◽

Vol 18 (1) ◽

Author(s):

Ella Yeor-Davidi ◽

Marina Zverzhinetsky ◽

Vadim Krivitsky ◽

Fernando Patolsky

Keyword(s):

Real Time ◽

Metabolic Activity ◽

Bacterial Biofilms ◽

Real Time Monitoring

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Real-time Monitoring of Bacterial Biofilms Metabolic Activity by a Redox-Reactive Nanosensors Array

10.21203/rs.3.rs-20969/v1 ◽

2020 ◽

Author(s):

Ella Yeor Davidi ◽

Marina Zverzhinetsky ◽

Vadim Krivitsky ◽

Fernando Patolsky

Keyword(s):

Ionic Strength ◽

Real Time ◽

Metabolic Activity ◽

High Ionic Strength ◽

Bacterial Biofilm ◽

Bacterial Biofilms ◽

Proper Treatment ◽

Real Time Monitoring ◽

Waste Products ◽

Shed Light

Abstract Background: Bacterial biofilms are communities of surface-associated microorganisms living in cellular clusters or micro-colonies, encapsulated in a complex matrix composed of an extracellular polymeric substance, separated by open water channels that act as a circulatory system that enable better diffusion of nutrients and easier removal of metabolic waste products. The monitoring of biofilms can provide important information on fundamental biofilm-related processes. That information can shed light on the bacterial processes and enable scientists to find ways of preventing future bacterial infections. Various approaches in use for biofilm analysis are based on microscopic, spectrochemical, electrochemical and piezoelectrical methods. All these methods provide significant progress in understanding the bio-process related to biofilm formation and eradication, nevertheless, the development of novel approaches for the real-time monitoring of biochemical, in particular metabolic activity, of bacterial species during the formation, life and eradication of biofilms is of great potential importance.Results: Here, detection and monitoring of the metabolic activity of bacterial biofilms in high-ionic-strength solutions were enabled as a result of novel surface modification by an active redox system, composed of 9,10-dihydroxyanthracene/9,10-anthraquinone, on the oxide layer of the SiNW, yielding a chemically-gated FET array. With the use of enzymatic reactions of oxidases, metabolites can be converted to and monitored by the nanosensors. Here, the successful detection of glucose consumption in high-ionic-strength solutions, such as bacterial media, without pre-processing of small volume samples under different conditions and treatments, has been demonstrated. The biofilms were treated with antibiotics differing in their mechanisms of action and were compared to untreated biofilms. Further examination of biofilms under antibiotic treatment with SiNW-FET devices could shed light on the bioprocess that occurs within the biofilm. Moreover, finding a proper treatment that eliminates the biofilm could be examined by the novel nanosensor array as a monitoring tool.Conclusions: To summarize, the combination of redox-reactive SiNW-FET devices with microfluidic techniques enables the performance of label-free, rapid, automated, multiplex and real-time noninvasive metabolites detection for the investigation of bacterial biofilms. This novel platform can be used as an extremely sensitive tool for detection and establishing medical solutions for bacterial-biofilm eradication and for finding a proper treatment to eliminate biofilm contaminations. Moreover, the sensing system can be used as a research tool for further understanding of the metabolic processes that occur within the bacterial biofilm population.

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Effects of Low Molecular Weight Fibrinogen Degradation Products on Lymphocytes, Macrophages and Kidney Cells in Cultures

10.1055/s-0039-1687221 ◽

1979 ◽

Author(s):

M. Kopeć ◽

W. Rosazkowski ◽

S. Szmigielski ◽

B. Gerdin ◽

T. Saldeen

Keyword(s):

Degradation Products ◽

Biological Activities ◽

Peritoneal Macrophages ◽

Rabbit Kidney ◽

Low Molecular Weight ◽

Kidney Cells ◽

Dependent Manner ◽

Dose Dependent ◽

Mouse Peritoneal Macrophages ◽

Molecular Sieving

Dialysable peptides of mw 2200 to 3500 D(LMW-FDP) were isolated from products of exhaustive proteolysis of fibrinogen by plasmin. LMW-FDP inhibited in a dose dependent manner incorporation of 3H-thymidme into cultures of human blood lymphocytes simulated with PHA, concanavalin A or bacterial lipopolisaccharide. These effects were inversely related to stimulation indices. Phagocytosis of 3H-thymidine labelled Staphylococcus aureus by mouse peritoneal macrophages was also inhibited by LMW-FDP. These peptides induced a cytotoxic effect on continuous line of rabbit kidney cells in cultures as manifested by inhibition of 86Rb and 3H-incorporation as well as by releese of 86Rb and 61Cr from prelabelled cells. Eight fractions obtained by molecular sieving of LMW - FDP on Bio-Gel P-6 columns were found to differ pronouncedly in biological activities.

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Real-time monitoring of the metabolic activity of periodontopathic bacteria

Journal of Microbiological Methods ◽

10.1016/j.mimet.2015.05.015 ◽

2015 ◽

Vol 115 ◽

pp. 22-26 ◽

Cited By ~ 8

Author(s):

Kazuko Ishiguro ◽

Jumpei Washio ◽

Keiichi Sasaki ◽

Nobuhiro Takahashi

Keyword(s):

Real Time ◽

Metabolic Activity ◽

Real Time Monitoring ◽

Periodontopathic Bacteria

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Real-time Monitoring of Bacterial Biofilms Metabolic Activity by a Redox-Reactive Nanosensors Array

10.21203/rs.3.rs-20969/v2 ◽

2020 ◽

Author(s):

Fernando Patolsky ◽

Vadim Krivitsky ◽

Marina Zverzhinetsky ◽

Ella Yeor Davidi

Keyword(s):

Ionic Strength ◽

Real Time ◽

Metabolic Activity ◽

High Ionic Strength ◽

Bacterial Biofilm ◽

Bacterial Biofilms ◽

Proper Treatment ◽

Real Time Monitoring ◽

Waste Products ◽

Shed Light

Abstract Background Bacterial biofilms are communities of surface-associated microorganisms living in cellular clusters or micro-colonies, encapsulated in a complex matrix composed of an extracellular polymeric substance, separated by open water channels that act as a circulatory system that enable better diffusion of nutrients and easier removal of metabolic waste products. The monitoring of biofilms can provide important information on fundamental biofilm-related processes. That information can shed light on the bacterial processes and enable scientists to find ways of preventing future bacterial infections. Various approaches in use for biofilm analysis are based on microscopic, spectrochemical, electrochemical, and piezoelectrical methods. All these methods provide significant progress in understanding the bio-process related to biofilm formation and eradication, nevertheless, the development of novel approaches for the real-time monitoring of biochemical, in particular metabolic activity, of bacterial species during the formation, life and eradication of biofilms is of great potential importance. Results Here, detection and monitoring of the metabolic activity of bacterial biofilms in high-ionic-strength solutions were enabled as a result of novel surface modification by an active redox system, composed of 9,10-dihydroxyanthracene/9,10-anthraquinone, on the oxide layer of the SiNW, yielding a chemically-gated FET array. With the use of enzymatic reactions of oxidases, metabolites can be converted to Common.EditSubmissionSteps.Transform.EquationText and monitored by the nanosensors. Here, the successful detection of glucose metabolites in high-ionic-strength solutions, such as bacterial media, without pre-processing of small volume samples under different conditions and treatments, has been demonstrated. The biofilms were treated with antibiotics differing in their mechanisms of action and were compared to untreated biofilms. Further examination of biofilms under antibiotic treatment with SiNW-FET devices could shed light on the bioprocess that occurs within the biofilm. Moreover, finding proper treatment that eliminates the biofilm could be examined by the novel nanosensor as a monitoring tool. Conclusions To summarize, the combination of redox-reactive SiNW-FET devices with micro-fluidic techniques enables the performance of rapid, automated, and real-time metabolite detection with the use of minimal sample size, noninvasively and label-free. This novel platform can be used as an extremely sensitive tool for detection and establishing medical solutions for bacterial-biofilm eradication and for finding a proper treatment to eliminate biofilm contaminations. Moreover, the sensing system can be used as a research tool for further understanding of the metabolic processes that occur within the bacterial biofilm population.

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