Low-Oxygen Pretreatment Enhances Endothelial Cell Growth and Retention Under Shear Stress

Abstract

Oxygen (O(2)) tension is an important factor that regulates endothelial cell (EC) growth and adhesion. We hypothesized that low-O(2) treatment of ECs improves the endothelialization and cell retention upon physiologically relevant perfusion flow, due to enhanced cell proliferation and extracellular matrix (ECM) secretion. We assessed the effects of a low-O(2) tension of 5% O(2) upon growth and ECM production of human umbilical vein ECs (HUVECs), in comparison to their counterparts at 20% O(2) on poly(ethylene terephthalate) (PET) films. Low-O(2) pretreatment at 5% O(2) promoted HUVEC proliferation, ECM secretion, and intercellular adhesion. Cell retentions of the endothelialized PET films formed under 5% and 20% O(2) were analyzed by applying shear stress in the range of 5-20 dyn/cm(2) for up to 24 h under the O(2) of 12% and 20%, mimicking arterial and conventional experimental O(2), respectively. The 5% O(2)-pretreated samples exhibited significantly higher cell retention than their normoxic counterparts at high cell density (>30 x 10(3) cells/cm(2)) over extended exposure time (>12 h) when perfused under both 12% and 20% O(2). The endothelium formed under 5% O(2) maintained its ability to respond to perfusion flow by upregulating nitric oxide and prostacyclin production under both O(2) perfusion conditions. The results indicate that pretreatment at 5% O(2) is an effective strategy to enhance endothelialization of vascular grafts by promoting endothelium formation, cell retention, and function.