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Cell-based sensor system using L6 cells for broad band continuous pollutant monitoring in aquatic environments.

Kubisch R, Bohrn U, Fleischer M, Stütz E - Sensors (Basel) (2012)

Bottom Line: In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown.A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts).In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Biology-Biotechnology, Department of Pharmacy, Center for Drug Research, Ludwig-Maximilian-University Munich, Munich, Germany. rebekka.kubisch@cup.uni-muenchen.de

ABSTRACT
Pollution of drinking water sources represents a continuously emerging problem in global environmental protection. Novel techniques for real-time monitoring of water quality, capable of the detection of unanticipated toxic and bioactive substances, are urgently needed. In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown. Readout parameters of the cells were the acidification (metabolism), oxygen consumption (respiration) and impedance (morphology) of the cells. A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts). The cytotoxicity or cellular effects induced by inorganic ions (Ni(2+) and Cu(2+)) can be detected with the metabolic parameters acidification and respiration down to 0.5 mg/L, whereas the detection limit for other substances like nicotine and acetaminophen are rather high, in the range of 0.1 mg/L and 100 mg/L. In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances. The results support the paradigm change from single substance detection to the monitoring of overall toxicity.

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Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with nicotine in the Bionas 2500 Analyzing System. Vehicle control (black), 1,000 mg/L (red), 500 mg/L (blue), 250 mg/L (green), 100 mg/L (yellow). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of nicotine exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with nicotine for 24 h.
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f7-sensors-12-03370: Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with nicotine in the Bionas 2500 Analyzing System. Vehicle control (black), 1,000 mg/L (red), 500 mg/L (blue), 250 mg/L (green), 100 mg/L (yellow). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of nicotine exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with nicotine for 24 h.

Mentions: In the Bionas analyzing system, nicotine shows a dose dependent decrease of impedance at high concentrations (>250 mg/L; see Figure 7(a)). This morphological disturbance is only slightly recovered after 3 h of incubation in clean running medium incubation. According to the literature, breakdown of the membrane integrity due to peroxidized lipid components in the cell membrane might be an explanation for the impedance changes. Concentrations up to 250 mg/L lead to a strong increase of cellular acidification while this trend is inverted at higher concentrations of 500 and 1,000 mg/L, resulting in a decreased metabolism (see Figure 7(b)). In contrast, the results of the respiration measurements are all oriented into the same direction (see Figure 7(c)). The endpoint values of the changes relative to the control after 15 h of incubation show that high concentrations of nicotine (500 and 1,000 mg/L) decrease the respiration down to 50–70% (see Figure 7(d)). One can assume that the generation of ROS causes damage in the respiration system of the cells. The BrdU assay results show a constant decrease of the proliferative activity at nicotine concentration range from ∼100 mg/L up to 12.5 g/L (see Figure 7(e)). The respiration rates obtained with the Bionas system and to a minor amount the impedance values are in accordance with the BrdU results as they show as well a continuously decrease at nicotine concentrations from 100 mg/L to 1,000 mg/L.


Cell-based sensor system using L6 cells for broad band continuous pollutant monitoring in aquatic environments.

Kubisch R, Bohrn U, Fleischer M, Stütz E - Sensors (Basel) (2012)

Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with nicotine in the Bionas 2500 Analyzing System. Vehicle control (black), 1,000 mg/L (red), 500 mg/L (blue), 250 mg/L (green), 100 mg/L (yellow). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of nicotine exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with nicotine for 24 h.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3376625&req=5

f7-sensors-12-03370: Time course of the impedance (a), acidification (b) and respiration (c) of L6 rat skeletal muscle cells during 24 h incubation with nicotine in the Bionas 2500 Analyzing System. Vehicle control (black), 1,000 mg/L (red), 500 mg/L (blue), 250 mg/L (green), 100 mg/L (yellow). Phases with running medium (RM) represent culture medium treatment without added substances. (d) Percent of control of impedance, respiration and acidification endpoint values measured after 15 h of nicotine exposure (n = 3). (e) Results of BrdU assay of L6 cells incubated with nicotine for 24 h.
Mentions: In the Bionas analyzing system, nicotine shows a dose dependent decrease of impedance at high concentrations (>250 mg/L; see Figure 7(a)). This morphological disturbance is only slightly recovered after 3 h of incubation in clean running medium incubation. According to the literature, breakdown of the membrane integrity due to peroxidized lipid components in the cell membrane might be an explanation for the impedance changes. Concentrations up to 250 mg/L lead to a strong increase of cellular acidification while this trend is inverted at higher concentrations of 500 and 1,000 mg/L, resulting in a decreased metabolism (see Figure 7(b)). In contrast, the results of the respiration measurements are all oriented into the same direction (see Figure 7(c)). The endpoint values of the changes relative to the control after 15 h of incubation show that high concentrations of nicotine (500 and 1,000 mg/L) decrease the respiration down to 50–70% (see Figure 7(d)). One can assume that the generation of ROS causes damage in the respiration system of the cells. The BrdU assay results show a constant decrease of the proliferative activity at nicotine concentration range from ∼100 mg/L up to 12.5 g/L (see Figure 7(e)). The respiration rates obtained with the Bionas system and to a minor amount the impedance values are in accordance with the BrdU results as they show as well a continuously decrease at nicotine concentrations from 100 mg/L to 1,000 mg/L.

Bottom Line: In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown.A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts).In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Biology-Biotechnology, Department of Pharmacy, Center for Drug Research, Ludwig-Maximilian-University Munich, Munich, Germany. rebekka.kubisch@cup.uni-muenchen.de

ABSTRACT
Pollution of drinking water sources represents a continuously emerging problem in global environmental protection. Novel techniques for real-time monitoring of water quality, capable of the detection of unanticipated toxic and bioactive substances, are urgently needed. In this study, the applicability of a cell-based sensor system using selected eukaryotic cell lines for the detection of aquatic pollutants is shown. Readout parameters of the cells were the acidification (metabolism), oxygen consumption (respiration) and impedance (morphology) of the cells. A variety of potential cytotoxic classes of substances (heavy metals, pharmaceuticals, neurotoxins, waste water) was tested with monolayers of L6 cells (rat myoblasts). The cytotoxicity or cellular effects induced by inorganic ions (Ni(2+) and Cu(2+)) can be detected with the metabolic parameters acidification and respiration down to 0.5 mg/L, whereas the detection limit for other substances like nicotine and acetaminophen are rather high, in the range of 0.1 mg/L and 100 mg/L. In a close to application model a real waste water sample shows detectable signals, indicating the existence of cytotoxic substances. The results support the paradigm change from single substance detection to the monitoring of overall toxicity.

Show MeSH
Related in: MedlinePlus