Optimization of Electric Field Parameters for HT29 Cell Line towards Wound Healing Application

Abstract

Objective: The aim of this study is to optimize the electric field parameter for HT-29 cells line towards wound healing application. Methods: Cells were harvested when they reached 70% to 80% confluent. Afterward, 800 μl of the cells suspension were poured into a 4 mm cuvette and placed in the BTX ECM 830 Electroporator chamber. Voltages in the range of 80 V to 800 V and pulse durations in the range of 100 μs to 10 ms were used in electroporating the cells. After treatment, 300 μl of the cells suspension from each sample were seeded in a well of 6-wells plates each containing 2 ml of pre-warm complete growth medium and incubated at 37oC and 5% CO2 for 48 hours. Findings: The results obtained in this research quantitatively reveal the dependence of cell proliferation on electrical parameters. 200, 400, 600 and 800 V/cm electric field strengths at duration of 500 μs showed 42%, 63%, 86% and 36% increase in proliferation rate after 48 hours in culture as compared to initial plating densities, respectively. The control group and 1000 V/cm at 100 μs showed 48% and 33% increase in proliferation rate respectively as compared to initial seeding density. While electric field strengths of 1200-2000 V/cm at 200 V/cm interval and at 100 μs duration, revealed a decrease in proliferation rate by 4%, 15%, 18%, 23% and 26% respectively as compared to initial seeding density. Applications: It was found that 600 V/cm at 500 μs induced the highest proliferation rate (86%) as compared to initial seeding density after 48 hours of culture (that is 38% greater than the control group). This study can further be investigated for the use of the pulse electric field in facilitating drug-free wound healing process.