Investigation of Pulse electric field effect on HeLa cells alignment properties on extracellular matrix protein patterned surface

Muhammad Mahadi Abdul Jamil,Morgan C T Denyer,Radzi Ambar,Mansour Youseffi,Nur Adilah Abd Rahman,Mohamed Ahmed Milad Zaltum,Masoud Mozafari,Farshid Sefat,F Javed

doi:10.1088/1742-6596/1019/1/012018

Abstract

Cell behavior in terms of adhesion, orientation and guidance, on extracellular matrix (ECM) molecules including collagen, fibronectin and laminin can be examined using micro contact printing (MCP). These cell adhesion proteins can direct cellular adhesion, migration, differentiation and network formation in-vitro. This study investigates the effect of micro-contact printed ECM protein, namely fibronectin, on alignment and morphology of HeLa cells cultured in-vitro. Fibronectin was stamped on plain glass cover slips to create patterns of 25µm, 50µm and 100µm width. However, HeLa cells seeded on 50µm induced the best alignment on fibronectin pattern (7.66° ±1.55SD). As a consequence of this, 50µm wide fibronectin pattern was used to see how fibronectin induced cell guidance of HeLa cells was influenced by 100µs and single pulse electric fields (PEF) of 1kV/cm. The results indicates that cells aligned more under pulse electric field exposure (2.33° ±1.52SD) on fibronectin pattern substrate. Thus, PEF usage on biological cells would appear to enhance cell surface attachment and cell guidance. Understanding this further may have applications in enhancing tissue graft generation and potentially wound repair.

Highlights

Micro-contact printing (MCP) is a procedure that allows a surface to be well-designed with a material like extra cellular matrix (ECM) protein, in a clear outline [1,2,3,4]
When the HeLa cells reached 80-90% confluence, media was removed from the tissue culture flask, 3ml of phosphate buffered saline (PBS) was added in the flask to wash the cells, which was removed after few seconds. 2 ml of TryPle Express from GIBCO was added to detach the cells from bottom of the flask
Micro contact printing (MCP) technique The stamp consisted of ridges with 25μm, 50μm and 100μm width fibronectin coated tracks separated by 25μm, 50μm and 100μm width uncoated tracks

Summary

Introduction

Micro-contact printing (MCP) is a procedure that allows a surface to be well-designed with a material like extra cellular matrix (ECM) protein, in a clear outline [1,2,3,4]. Published under licence by IOP Publishing Ltd orientation and guidance, on ECM molecules such as fibronectin, collagen and laminin can be examined using MCP [5,6] These cell adhesion proteins can guide cellular migration, adhesion and network establishment in-vitro [7]. Fibronectin attach to collagen, fibroblasts in the ECM and the plasma membrane of the cells and its high molecular weight protein. Formation of detailed surface designs on glass, metal or polymer might allow further clarification about the elements that control cell attachment, cell growth, variation and molecular signaling paths on material surfaces [24,25]. The aims of this study are to characterize HeLa cell guidance on micro contact printed fibronectin patterns and to determine how that guidance is modified by pulse electric field

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Conclusion