Cell biology

Ecology

Biology

Endothelial stem cell

Chromatography

Reactive oxygen species

Extracellular matrix

Biochemistry

Food science

Substrate (aquarium)

Matrix (chemical analysis)

Organic chemistry

Laminin

Oxygen

Pyrogallol

In vitro

Oxidative phosphorylation

Oxidative stress

Chemistry

Cell

Ascorbic acid

Biophysics

Artificial Organs

Abstract: Anchorage‐dependent cells in culture attach initially to proteins adsorbed to the culture substrate from the medium, and produce and deposit a subcellular matrix during the course of the cultivation. The aim of this study was to determine whether the concentration of O 2 in the culture atmosphere affects the accumulation of type IV collagen and laminin under human endothelial‐cell monolayers. Enzyme‐linked immunoassays on decellularized polystyrene substrates showed less type IV collagen, but not less laminin, under cells incubated in the standard atmosphere (5% CO 2 in air, i.e., ∼20% O 2 ) compared to an atmosphere of 5% O 2 and 5% CO 2 in N 2 . Type IV collagen accumulation was inhibited via oxidative stress, because the inhibitory effect of 20% O 2 was antagonized by antioxidant ascorbic acid, and mimicked by prooxidant pyrogallol and exogenous H 2 O 2 . Measurements of endogenous H 2 O 2 accumulation demonstrated that endothelial cells partially adapt to the high O 2 concentration. These results may have implications in endothelium modeling in vitro and in engineering of endothelial cell sheets and endothelialized vascular grafts.

Oxygen Reduces Accumulation of Type IV Collagen in Endothelial Cell Subcellular Matrix via Oxidative Stress