Effects of Stiffness on Endothelial Cells Behavior

Authors: Forough Ataollahi, Belinda Pingguan-Murphy, Wan Abu Bakar Wan Abas, Noor Azuan Abu Osman

Abstract:

Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al₂O₃) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al₂O₃ at curing temperature 50˚C for 4h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.

Keywords: Bovine aortic endothelial cells, extra cellular matrix, proliferation, stiffness.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1090691

References:

[1] H.-I. Chang and Y. Wang, "Cell responses to surface and architecture of tissue engineering scaffolds". Regenerative Medicine and Tissue Engineering—Cells and Biomaterials, In Tech: Rijeka, Croatia, 2011, pp. 569-588.
[2] E.J. Gang, et al., "In vitro mesengenic potential of human umbilical cord blood-derived mesenchymal stem cells". Biochemical and Biophysical Research Communications, 2004, pp. 102-108.
[3] M. Ponticos, et al., "Regulation of collagen type I in vascular smooth muscle cells by competition between Nkx2.5 and deltaEF1/ZEB1". Molecular and cellular biology, 2004, pp. 6151-6161.
[4] Y.-T. Yeh, et al., "Matrix Stiffness Regulates Endothelial Cell Proliferation through Septin 9". PLoS ONE, 2012, pp. e46889.
[5] D.B. Cines, et al., "Endothelial Cells in Physiology and in the Pathophysiology of Vascular Disorders". Blood, 1998, pp. 3527-3561.
[6] R.J. Pelham and Y.-l. Wang, "Cell locomotion and focal adhesions are regulated by substrate flexibility". Proceedings of the National Academy of Sciences, 1997, pp. 13661-13665.
[7] K. Ramaswamy, et al., "Substrate stiffening promotes endothelial monolayer disruption through enhanced physical forces". American Journal of Physiology - Cell Physiolog, 2010, pp. C146-C154.
[8] P.S. Tan and S.H. Teoh, "Effect of stiffness of polycaprolactone (PCL) membrane on cell proliferation". Materials Science and Engineering: C, 2007, pp. 304-308.
[9] T. Yeung, et al., "Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion". Cell Motility and the Cytoskeleton, 2005, pp. 24-34.
[10] H. Takeda, et al., "Bi-phasic activation of eNOS in response to uni-axial cyclic stretch is mediated by differential mechanisms in BAECs". Life Sciences, 2006, pp. 233-239.
[11] J.N. Lee, et al., "Compatibility of Mammalian Cells on Surfaces of Poly(dimethylsiloxane)". Langmuir, 2004, pp. 11684-11691.
[12] Z. Weijia, et al., "Studying Cancer Stem Cell Dynamics on PDMS Surfaces for Microfluidics Device Design". Scientific Reports, 2013, pp.
[13] B. Leukers, et al., "Biocompatibility of ceramic scaffolds for bone replacement made by 3D printing". Materialwissenschaft und Werkstofftechnik, 2005, pp. 781-787.
[14] S. Jantová, et al., "Biocompatibility and cytotoxicity of bioglass-ceramic composite with various P2O5 content in Li2O–SiO2–CaO–CaF2–P2O5 system on fibroblast cell lines". Acta Chimica Slovaca, 2011, pp. 15-30.
[15] R. Sunyer, et al., "Fabrication of Hydrogels with Steep Stiffness Gradients for Studying Cell Mechanical Response". PLoS ONE, 2012, pp. e46107.
[16] A.J. Engler, et al., "Matrix Elasticity Directs Stem Cell Lineage Specification". Cell, 2006, pp. 677-689.