Abstract
Understanding signals in the microenvironment that regulate endothelial cell behavior are important in tissue engineering. Although many studies have examined the cellular effects of nanotopography, no study has investigated the functional regulation of human endothelial cells grown on nano-sized gradient hole substrate. We examined the cellular response of human umbilical vein endothelial cells (HUVECs) by using a gradient nanohole substrate (GHS) with three different types of nanohole patterns (HP): which diameters were described in HP1, 120–200 nm; HP2, 200–280 nm; HP3, 280–360 nm. In results, HP2 GHS increased the attachment and proliferation of HUVECs. Also, gene expression of focal adhesion markers in HUVECs was significantly increased on HP2 GHS. In vitro tube formation assay showed the enhancement of tubular network formation of HUVECs after priming on GHS compared to Flat. Furthermore, leukocyte adhesion was also reduced in the HUVECs in a hole-diameter dependent manner. To summarize, optimal proliferations with reduced leukocyte adhesion of HUVECs were achieved by gradient nanohole substrate with 200–280 nm-sized holes.
Highlights
Endothelial cells are the essential component of blood vessels which are in direct contact with vascular smooth muscles and connective tissues[1]
Recent reports have shown that the nano-scale surface topography plays a critical role in changing responses of various types of stem cells and adult cells, and for example, the mesenchymal stem cells (MSC) differentiation into osteoblasts or adipocytes was influenced by the substrate stiffness and nano-sized pattern[10]
This study revealed that; (i) human umbilical vein endothelial cells (HUVECs) showed the highest attachment and proliferation capacity on HP2 (200– 280 nm) gradient nanohole substrate (GHS), (ii) HP2 GHS was determined to be the optimal diameter for extending greater tube forming potential to HUVECs and (iii) HP2 GHS reduce the leukocyte adhesion compared to Flat
Summary
Endothelial cells are the essential component of blood vessels which are in direct contact with vascular smooth muscles and connective tissues[1]. Recent reports have shown that the nano-scale surface topography plays a critical role in changing responses of various types of stem cells and adult cells, and for example, the mesenchymal stem cells (MSC) differentiation into osteoblasts or adipocytes was influenced by the substrate stiffness and nano-sized pattern[10]. Another study reported that nanopattern substrate could provide a regulatory signal to influence the human embryonic stem cell responses including cell morphology, adhesion, proliferation, and self-renewal potential[11]. It is the first time to adopt the gradient nanohole substrate (GHS) to screen the optimal growth condition for human umbilical vein endothelial cells (HUVECs), and GHS provides different conditions of nano-scale environment for evaluating the endothelial cell response. This study hypothesized that GHS would play a pivotal role in the HUVECs culture system and mimic endothelium microenvironment in vitro
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