Electroporation-mediated delivery of molecules to model intestinal epithelia.
This study was conducted to determine if electroporation can deliver membrane-impermeant molecules intracellularly to intact, physiologically competent monolayers that mimic the intestinal epithelium. In addition, the long-term effects of electroporation on these monolayers were studied to determine the kinetics with which monolayers recover barrier function. Caco-2 and T84 cells were electroporated as monolayers using calcein and fluorescein-labeled bovine serum albumin as marker molecules for measuring delivery into cells. Confocal microscopy and flow cytometry were used, respectively, to visualize and quantify uptake of these molecules. Transepithelial resistance was used as a measure of physiologic barrier function. We found that intracellular uptake of calcein and bovine serum albumin occurred uniformly throughout both types of model epithelia and increased as a function of voltage, pulse length, and pulse number. There was no significant difference in uptake resulting from single and multiple pulses of the same total exposure time. We also observed that monolayers exposed to electroporation that induced uptake of up to 10(6) molecules/cell were able to recover normal barrier function within one day. These findings suggest that electroporation may be useful for intracellular delivery into monolayers to study epithelial biology and, possibly, for drug delivery to intestinal epithelium.
Ghartey-Tagoe
Esi B
EB
The Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University, Georgia Institute of Technology, Atlanta, GA 30332, USA.
Morgan
Jeremy S
JS
Ahmed
Kashif
K
Neish
Andrew S
AS
Prausnitz
Mark R
MR
D000818:Animals; D015415:Biomarkers; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D018274:Electroporation; D005434:Flow Cytometry; D019793:Fluorescein / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D018613:Microscopy, Confocal; D008954:Models, Biological; D010539:Permeability; D004364:Pharmaceutical Preparations / Q000008:administration & dosage* / Q000378:metabolism; D010599:Pharmacokinetics; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013997:Time Factors
D015415:Biomarkers; D005452:Fluoresceins; D004364:Pharmaceutical Preparations; D012710:Serum Albumin, Bovine; D019793:Fluorescein; C007740:fluorexon