Burn-activated neutrophils and tumor necrosis factor-α alter endothelial cell actin cytoskeleton and enhance monolayer permeability

Keith Wright, Fiemu Nwariaku, Nabil Halaihel, Carl Schulman, Helen L. Yin, Richard H. Turnage

Research output: Contribution to journalArticle

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Abstract

Background. This study examined the hypothesis that exposure of an endothelial cell (EC) monolayer to tumor necrosis factor-α (TNF-α) and that burn-activated neutrophils alter EC actin cytoskeleton and enhance the permeability of the monolayer. Methods. Neutrophils were harvested from rats that had undergone a 45 % surface area burn (BURN-neutrophil) or uninjured control rats. ECs were grown on polyester filters or fibronectin-coated glass slides and exposed for 4 hours to media, TNF-α (100 ng/mL), or TNF-α plus BURN-neutrophil or uninjured control rats (107 cells). Monolayer permeability was assessed by measuring the flux of albumin across the cells. EC surface area and microfilament number and length were determined by the staining of actin microfilaments with rhodamine phalloidin followed by fluorescent microscopy. Results. The amount of albumin that moved across the monolayer in response to TNF-α plus BURN-neutrophil was twice that of media alone (P < .05) or TNF-α alone (P < .05). The number and length of actin microfilaments in ECs exposed to TNF-α plus BURN-neutrophil were significantly less than that of cells exposed to media alone or TNF-α alone. Conclusions. These data are consistent with a hypothesis that TNF-α plus BURN-neutrophil affect endothelial monolayer permeability by altering EC actin cytoskeletal organization.

Original languageEnglish (US)
Pages (from-to)259-265
Number of pages7
JournalSurgery
Volume128
Issue number2
DOIs
StatePublished - 2000

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Actin Cytoskeleton
Burns
Permeability
Neutrophils
Endothelial Cells
Tumor Necrosis Factor-alpha
Albumins
Polyesters
Fibronectins
Glass
Actins
Microscopy
Staining and Labeling

ASJC Scopus subject areas

  • Surgery

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Burn-activated neutrophils and tumor necrosis factor-α alter endothelial cell actin cytoskeleton and enhance monolayer permeability. / Wright, Keith; Nwariaku, Fiemu; Halaihel, Nabil; Schulman, Carl; Yin, Helen L.; Turnage, Richard H.

In: Surgery, Vol. 128, No. 2, 2000, p. 259-265.

Research output: Contribution to journalArticle

Wright, Keith ; Nwariaku, Fiemu ; Halaihel, Nabil ; Schulman, Carl ; Yin, Helen L. ; Turnage, Richard H. / Burn-activated neutrophils and tumor necrosis factor-α alter endothelial cell actin cytoskeleton and enhance monolayer permeability. In: Surgery. 2000 ; Vol. 128, No. 2. pp. 259-265.
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T1 - Burn-activated neutrophils and tumor necrosis factor-α alter endothelial cell actin cytoskeleton and enhance monolayer permeability

AU - Wright, Keith

AU - Nwariaku, Fiemu

AU - Halaihel, Nabil

AU - Schulman, Carl

AU - Yin, Helen L.

AU - Turnage, Richard H.

PY - 2000

Y1 - 2000

N2 - Background. This study examined the hypothesis that exposure of an endothelial cell (EC) monolayer to tumor necrosis factor-α (TNF-α) and that burn-activated neutrophils alter EC actin cytoskeleton and enhance the permeability of the monolayer. Methods. Neutrophils were harvested from rats that had undergone a 45 % surface area burn (BURN-neutrophil) or uninjured control rats. ECs were grown on polyester filters or fibronectin-coated glass slides and exposed for 4 hours to media, TNF-α (100 ng/mL), or TNF-α plus BURN-neutrophil or uninjured control rats (107 cells). Monolayer permeability was assessed by measuring the flux of albumin across the cells. EC surface area and microfilament number and length were determined by the staining of actin microfilaments with rhodamine phalloidin followed by fluorescent microscopy. Results. The amount of albumin that moved across the monolayer in response to TNF-α plus BURN-neutrophil was twice that of media alone (P < .05) or TNF-α alone (P < .05). The number and length of actin microfilaments in ECs exposed to TNF-α plus BURN-neutrophil were significantly less than that of cells exposed to media alone or TNF-α alone. Conclusions. These data are consistent with a hypothesis that TNF-α plus BURN-neutrophil affect endothelial monolayer permeability by altering EC actin cytoskeletal organization.

AB - Background. This study examined the hypothesis that exposure of an endothelial cell (EC) monolayer to tumor necrosis factor-α (TNF-α) and that burn-activated neutrophils alter EC actin cytoskeleton and enhance the permeability of the monolayer. Methods. Neutrophils were harvested from rats that had undergone a 45 % surface area burn (BURN-neutrophil) or uninjured control rats. ECs were grown on polyester filters or fibronectin-coated glass slides and exposed for 4 hours to media, TNF-α (100 ng/mL), or TNF-α plus BURN-neutrophil or uninjured control rats (107 cells). Monolayer permeability was assessed by measuring the flux of albumin across the cells. EC surface area and microfilament number and length were determined by the staining of actin microfilaments with rhodamine phalloidin followed by fluorescent microscopy. Results. The amount of albumin that moved across the monolayer in response to TNF-α plus BURN-neutrophil was twice that of media alone (P < .05) or TNF-α alone (P < .05). The number and length of actin microfilaments in ECs exposed to TNF-α plus BURN-neutrophil were significantly less than that of cells exposed to media alone or TNF-α alone. Conclusions. These data are consistent with a hypothesis that TNF-α plus BURN-neutrophil affect endothelial monolayer permeability by altering EC actin cytoskeletal organization.

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