Single-step electrical field strength screening to determine electroporation induced transmembrane transport parameters.
The design of effective electroporation protocols for molecular delivery applications requires the determination of transport parameters including diffusion coefficient, membrane resealing, and critical electric field strength for electroporation. The use of existing technologies to determine these parameters is time-consuming and labor-intensive, and often results in large inconsistencies in parameter estimation due to variations in the protocols and setups. In this work, we suggest using a set of concentric electrodes to screen a full range of electric field strengths in a single test to determine the electroporation-induced transmembrane transport parameters. Using Calcein as a fluorescent probe, we developed analytical methodology to determine the transport parameters based on the electroporation-induced pattern of fluorescence loss from cells. A monolayer of normal human dermal fibroblast (NHDF) cells were pre-loaded with Calcein and electroporated with an applied voltage of 750V with 10 and 50 square pulses with 50μs duration. Using our analytical model, the critical electric field strength for electroporation was found for the 10 and 50 pulses experiments. An inverse correlation between the field strength and the molecular transport time decay constant, and a direct correlation between field strength and the membrane permeability were observed. The results of this work can simplify the development of electroporation-assisted technologies for research and therapies.
Department of Computer Science, Tel Aviv University, Israel.
Gadi
G
Blumrosen
The Center for Engineering in Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, United States.
Alireza
A
Abazari
The Center for Engineering in Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, United States; Porter School of Environmental Studies, Tel Aviv University, Israel. Electronic address: agolberg@tauex.tau.ac.il.
Alexander
A
Golberg
The Center for Engineering in Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, United States; Department of Biomedical Engineering, Rutgers University, Piscataway, NJ 08854., United States. Electronic address: ireis@sbi.org.
Martin L
ML
Yarmush
The Center for Engineering in Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, United States.
Mehmet
M
Toner
D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D002478:Cells, Cultured; D055590:Electromagnetic Phenomena; D018274:Electroporation; D005347:Fibroblasts / Q000737:chemistry* / Q000378:metabolism; D006801:Humans
Cell membrane permeability; Concentric electrode system; Electroporation; High-throughput experiments; Transport parameters