Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions.
<h4>Background</h4>Sensitization requires exposure to an allergen with subsequent production of a "danger "signal. In the skin, keratinocytes are the main producers of these signals.<h4>Objective</h4>To compare dose- and time-effects of cobalt on the viability of an...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2021-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0252159 |
| _version_ | 1819009123762569216 |
|---|---|
| author | Maria Klasson Magnus Lindberg Eva Särndahl Håkan Westberg Ing-Liss Bryngelsson Kedeye Tuerxun Alexander Persson |
| author_facet | Maria Klasson Magnus Lindberg Eva Särndahl Håkan Westberg Ing-Liss Bryngelsson Kedeye Tuerxun Alexander Persson |
| author_sort | Maria Klasson |
| collection | DOAJ |
| description | <h4>Background</h4>Sensitization requires exposure to an allergen with subsequent production of a "danger "signal. In the skin, keratinocytes are the main producers of these signals.<h4>Objective</h4>To compare dose- and time-effects of cobalt on the viability of and cytokine release from HaCaT cells cultured at low or high calcium.<h4>Method</h4>To model two separate states of differentiation of keratinocytes, HaCaT cells were cultured under low or high calcium conditions. HaCaT were exposed to different concentrations of cobalt chloride (10 μm to 5 mM) over time (30 minutes- 48 hours). Cell viability was measured with the Cell-Titer Blue Viability assay. Cytokine production was measured using a bead-based immunoassay and flow cytometry. Gene expression was quantified using qPCR. Data was analyzed by ANOVA and linear mixed model.<h4>Results</h4>Viability of the cells was dose- and time-dependent. A linear mixed statistical model showed that cobalt exposure induces increase in IL-6, CXCL8 and CCL2 production over time and whereas increase of IL-6 and a decrease of CCL2 was associated with increasing cobalt chloride concentrations. When comparing the cells incubated under high and low calcium conditions, the more differentiated cells in the high concentration were found to exert a stronger response in terms of IL-6 release.<h4>Conclusions</h4>Our data suggest that cobalt chloride triggered an alarm system in HaCaT cells, and proinflammatory cytokines/chemokines were secreted in a dose- and time-dependent manner. When high and low calcium incubations were compared, the difference was seen only for IL-6. These findings indicate that the effect of cobalt chloride on cell toxicity occurs throughout the living epidermis. |
| first_indexed | 2024-12-21T00:51:22Z |
| format | Article |
| id | doaj.art-a905b48e4f3c4f758a743e2c5eefc2a1 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T00:51:22Z |
| publishDate | 2021-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-a905b48e4f3c4f758a743e2c5eefc2a12022-12-21T19:21:24ZengPublic Library of Science (PLoS)PLoS ONE1932-62032021-01-01166e025215910.1371/journal.pone.0252159Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions.Maria KlassonMagnus LindbergEva SärndahlHåkan WestbergIng-Liss BryngelssonKedeye TuerxunAlexander Persson<h4>Background</h4>Sensitization requires exposure to an allergen with subsequent production of a "danger "signal. In the skin, keratinocytes are the main producers of these signals.<h4>Objective</h4>To compare dose- and time-effects of cobalt on the viability of and cytokine release from HaCaT cells cultured at low or high calcium.<h4>Method</h4>To model two separate states of differentiation of keratinocytes, HaCaT cells were cultured under low or high calcium conditions. HaCaT were exposed to different concentrations of cobalt chloride (10 μm to 5 mM) over time (30 minutes- 48 hours). Cell viability was measured with the Cell-Titer Blue Viability assay. Cytokine production was measured using a bead-based immunoassay and flow cytometry. Gene expression was quantified using qPCR. Data was analyzed by ANOVA and linear mixed model.<h4>Results</h4>Viability of the cells was dose- and time-dependent. A linear mixed statistical model showed that cobalt exposure induces increase in IL-6, CXCL8 and CCL2 production over time and whereas increase of IL-6 and a decrease of CCL2 was associated with increasing cobalt chloride concentrations. When comparing the cells incubated under high and low calcium conditions, the more differentiated cells in the high concentration were found to exert a stronger response in terms of IL-6 release.<h4>Conclusions</h4>Our data suggest that cobalt chloride triggered an alarm system in HaCaT cells, and proinflammatory cytokines/chemokines were secreted in a dose- and time-dependent manner. When high and low calcium incubations were compared, the difference was seen only for IL-6. These findings indicate that the effect of cobalt chloride on cell toxicity occurs throughout the living epidermis.https://doi.org/10.1371/journal.pone.0252159 |
| spellingShingle | Maria Klasson Magnus Lindberg Eva Särndahl Håkan Westberg Ing-Liss Bryngelsson Kedeye Tuerxun Alexander Persson Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions. PLoS ONE |
| title | Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions. |
| title_full | Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions. |
| title_fullStr | Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions. |
| title_full_unstemmed | Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions. |
| title_short | Dose- and time-dependent changes in viability and IL-6, CXCL8 and CCL2 production by HaCaT-cells exposed to cobalt. Effects of high and low calcium growth conditions. |
| title_sort | dose and time dependent changes in viability and il 6 cxcl8 and ccl2 production by hacat cells exposed to cobalt effects of high and low calcium growth conditions |
| url | https://doi.org/10.1371/journal.pone.0252159 |
| work_keys_str_mv | AT mariaklasson doseandtimedependentchangesinviabilityandil6cxcl8andccl2productionbyhacatcellsexposedtocobalteffectsofhighandlowcalciumgrowthconditions AT magnuslindberg doseandtimedependentchangesinviabilityandil6cxcl8andccl2productionbyhacatcellsexposedtocobalteffectsofhighandlowcalciumgrowthconditions AT evasarndahl doseandtimedependentchangesinviabilityandil6cxcl8andccl2productionbyhacatcellsexposedtocobalteffectsofhighandlowcalciumgrowthconditions AT hakanwestberg doseandtimedependentchangesinviabilityandil6cxcl8andccl2productionbyhacatcellsexposedtocobalteffectsofhighandlowcalciumgrowthconditions AT inglissbryngelsson doseandtimedependentchangesinviabilityandil6cxcl8andccl2productionbyhacatcellsexposedtocobalteffectsofhighandlowcalciumgrowthconditions AT kedeyetuerxun doseandtimedependentchangesinviabilityandil6cxcl8andccl2productionbyhacatcellsexposedtocobalteffectsofhighandlowcalciumgrowthconditions AT alexanderpersson doseandtimedependentchangesinviabilityandil6cxcl8andccl2productionbyhacatcellsexposedtocobalteffectsofhighandlowcalciumgrowthconditions |