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Stretch and/or oxygen glucose deprivation (OGD) in an in vitro traumatic brain injury (TBI) model induces calcium alteration and inflammatory cascade.

Salvador E, Burek M, Förster CY - Front Cell Neurosci (2015)

Bottom Line: Furthermore, reduction of cell membrane integrity decreased tight junction proteins claudin-5 and occludin expression.Also, since cell damage requires an increased uptake of glucose, expression of glucose transporter glut1 was found to increase at the mRNA level after OGD.Astrocytes potentiate these effects on calcium level in cEND cells.

View Article: PubMed Central - PubMed

Affiliation: Klinik und Poliklinik für Anästhesiologie, Zentrum für Operative Medizin der Universität Würzburg Würzburg, Germany.

ABSTRACT
The blood-brain barrier (BBB), made up of endothelial cells of capillaries in the brain, maintains the microenvironment of the central nervous system. During ischemia and traumatic brain injury (TBI), cellular disruption leading to mechanical insult results to the BBB being compromised. Oxygen glucose deprivation (OGD) is the most commonly used in vitro model for ischemia. On the other hand, stretch injury is currently being used to model TBI in vitro. In this paper, the two methods are used alone or in combination, to assess their effects on cerebrovascular endothelial cells cEND in the presence or absence of astrocytic factors. Applying severe stretch and/or OGD to cEND cells in our experiments resulted to cell swelling and distortion. Damage to the cells induced release of lactate dehydrogenase enzyme (LDH) and nitric oxide (NO) into the cell culture medium. In addition, mRNA expression of inflammatory markers interleukin (I L)-6, IL-1α, chemokine (C-C motif) ligand 2 (CCL2) and tumor necrosis factor (TNF)-α also increased. These events could lead to the opening of calcium ion channels resulting to excitotoxicity. This could be demonstrated by increased calcium level in OGD-subjected cEND cells incubated with astrocyte-conditioned medium. Furthermore, reduction of cell membrane integrity decreased tight junction proteins claudin-5 and occludin expression. In addition, permeability of the endothelial cell monolayer increased. Also, since cell damage requires an increased uptake of glucose, expression of glucose transporter glut1 was found to increase at the mRNA level after OGD. Overall, the effects of OGD on cEND cells appear to be more prominent than that of stretch with regards to TJ proteins, NO, glut1 expression, and calcium level. Astrocytes potentiate these effects on calcium level in cEND cells. Combining both methods to model TBI in vitro shows a promising improvement to currently available models.

No MeSH data available.


Related in: MedlinePlus

Lactate dehydrogenase (LDH) enzyme release into the cell culture medium in the various treatment conditions. LDH generated after stretch, OGD, a combination of both and successive reoxygenation was measured. LDH was expressed as a percent of the total releasable LDH (LDH in media plus cells). Data are mean values ± SEM, n = 3. *, ¤, $ p < 0.05 compared to no stretch, stretched, and OGD, respectively. OGD, oxygen glucose deprivation; RO, re-oxygenation.
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Figure 3: Lactate dehydrogenase (LDH) enzyme release into the cell culture medium in the various treatment conditions. LDH generated after stretch, OGD, a combination of both and successive reoxygenation was measured. LDH was expressed as a percent of the total releasable LDH (LDH in media plus cells). Data are mean values ± SEM, n = 3. *, ¤, $ p < 0.05 compared to no stretch, stretched, and OGD, respectively. OGD, oxygen glucose deprivation; RO, re-oxygenation.

Mentions: LDH release of cells that were subjected to severe stretch and made to recover for 15 min was compared to cells subjected to OGD and cells that underwent both conditions. It was observed that stretch released significantly higher LDH compared to no stretch control as well as OGD. Meanwhile, the combination of stretch and OGD generated a significantly higher amount of LDH compared to no stretch control, stretched cells, and cells subjected to OGD (Figure 3).


Stretch and/or oxygen glucose deprivation (OGD) in an in vitro traumatic brain injury (TBI) model induces calcium alteration and inflammatory cascade.

Salvador E, Burek M, Förster CY - Front Cell Neurosci (2015)

Lactate dehydrogenase (LDH) enzyme release into the cell culture medium in the various treatment conditions. LDH generated after stretch, OGD, a combination of both and successive reoxygenation was measured. LDH was expressed as a percent of the total releasable LDH (LDH in media plus cells). Data are mean values ± SEM, n = 3. *, ¤, $ p < 0.05 compared to no stretch, stretched, and OGD, respectively. OGD, oxygen glucose deprivation; RO, re-oxygenation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4543908&req=5

Figure 3: Lactate dehydrogenase (LDH) enzyme release into the cell culture medium in the various treatment conditions. LDH generated after stretch, OGD, a combination of both and successive reoxygenation was measured. LDH was expressed as a percent of the total releasable LDH (LDH in media plus cells). Data are mean values ± SEM, n = 3. *, ¤, $ p < 0.05 compared to no stretch, stretched, and OGD, respectively. OGD, oxygen glucose deprivation; RO, re-oxygenation.
Mentions: LDH release of cells that were subjected to severe stretch and made to recover for 15 min was compared to cells subjected to OGD and cells that underwent both conditions. It was observed that stretch released significantly higher LDH compared to no stretch control as well as OGD. Meanwhile, the combination of stretch and OGD generated a significantly higher amount of LDH compared to no stretch control, stretched cells, and cells subjected to OGD (Figure 3).

Bottom Line: Furthermore, reduction of cell membrane integrity decreased tight junction proteins claudin-5 and occludin expression.Also, since cell damage requires an increased uptake of glucose, expression of glucose transporter glut1 was found to increase at the mRNA level after OGD.Astrocytes potentiate these effects on calcium level in cEND cells.

View Article: PubMed Central - PubMed

Affiliation: Klinik und Poliklinik für Anästhesiologie, Zentrum für Operative Medizin der Universität Würzburg Würzburg, Germany.

ABSTRACT
The blood-brain barrier (BBB), made up of endothelial cells of capillaries in the brain, maintains the microenvironment of the central nervous system. During ischemia and traumatic brain injury (TBI), cellular disruption leading to mechanical insult results to the BBB being compromised. Oxygen glucose deprivation (OGD) is the most commonly used in vitro model for ischemia. On the other hand, stretch injury is currently being used to model TBI in vitro. In this paper, the two methods are used alone or in combination, to assess their effects on cerebrovascular endothelial cells cEND in the presence or absence of astrocytic factors. Applying severe stretch and/or OGD to cEND cells in our experiments resulted to cell swelling and distortion. Damage to the cells induced release of lactate dehydrogenase enzyme (LDH) and nitric oxide (NO) into the cell culture medium. In addition, mRNA expression of inflammatory markers interleukin (I L)-6, IL-1α, chemokine (C-C motif) ligand 2 (CCL2) and tumor necrosis factor (TNF)-α also increased. These events could lead to the opening of calcium ion channels resulting to excitotoxicity. This could be demonstrated by increased calcium level in OGD-subjected cEND cells incubated with astrocyte-conditioned medium. Furthermore, reduction of cell membrane integrity decreased tight junction proteins claudin-5 and occludin expression. In addition, permeability of the endothelial cell monolayer increased. Also, since cell damage requires an increased uptake of glucose, expression of glucose transporter glut1 was found to increase at the mRNA level after OGD. Overall, the effects of OGD on cEND cells appear to be more prominent than that of stretch with regards to TJ proteins, NO, glut1 expression, and calcium level. Astrocytes potentiate these effects on calcium level in cEND cells. Combining both methods to model TBI in vitro shows a promising improvement to currently available models.

No MeSH data available.


Related in: MedlinePlus