Studying the influence of electroporation on HT29 cell line interaction with micro-patterned extracellular matrix protein (fibronectin)

Abstract

Micro-contact printing (MCP) is a scheme that allows a substrate or surface to be functionalized freely with a protein of extracellular matrix (ECM), in a well-defined manner. MCP can be used to regulate cell adhesion geometry on a substrate and in controlling wound healing process. In this study, human colon cancer cell line, HT29 were grown on a micro-contact printed pattern of fibronectin with repeat gratings of 25µm, 50µm, and 100µm wide, for 48 hours. The cells alignments to the patterned substrates were then computed, where 0° means 100% alignment to the pattern. This was done with the purpose of finding those pattern that stimulated the best degree of cell alignment. The quantitative analysis of the study revealed that HT29 cells aligned most readily to the 50µm and 100µm pattern with a mean angle of alignment of 4.8°±1.5SD and 5.7°±1.4SD respectively. On the other hand, HT29 poorly aligned to 25um and the un-patterned fibronectin-coated substrate with a mean angle of alignment of 33.4°±8.4SD and 51.1°±9.5SD respectively. Furthermore, the 50µm stamp pattern was used to investigate the influence of electric field on the HT29 alignment to the fibronectin patterned substrate. Electroporated and non-electroporated HT29 cells were cultured on the 50µm patterned substrates for 48 hours. The result revealed that there is no significant difference in the degree of alignment between the electrically treated with a mean angle of 4.2°±1.4SD and the untreated cells with a mean angle of 4.8°±1.5SD. However electrically treated cell aligned faster (within 31 hours of seeding) as compared to the untreated cells that aligned after 44 hours of seeding. Hence, the result of the study revealed that Micro-contact printing technique together with pulse electric field could offer a potential faster method of controlling wound healing process.