title: Disruption of the actin cortex contributes to susceptibility of mammalian cells to nanosecond pulsed electric fields.
abstract: Nanosecond pulsed electric fields (nsPEFs) perturb membranes of cultured mammalian cells in a dose-dependent manner with different types of cells exhibiting characteristic survivability. Adherent cells appear more robust than non-adherent cells during whole-cell exposure. We hypothesize that cellular elasticity based upon the actin cytoskeleton is a contributing parameter, and the alteration of a cell's actin cortex will significantly affect viability upon nsPEF exposure. Chinese hamster ovary (CHO) cells that are (a) untreated, (b) treated with latrunculin A to inhibit actin polymerization, or (c) exposed to nsPEFs have been probed using atomic force microscopy (AFM) force-indentations. Exposure to 50 or 100 pulses of 10 ns duration and 150 kV/cm in a single dosage approximately lowers average CHO cell elastic modulus by half, whereas latrunculin lowers it more than 75%. Latrunculin pre-treatment disrupts the actin cortex enough that it negates cumulative damage by equally fractionated (i.e., two rounds of 50 pulses each, separated by 10 min) dosages of nsPEFs as seen in untreated and dimethyl sulfoxide (DMSO)-treated cells with propidium uptake, phosphatidylserine externalization, and 24 h viability according to MTT and CellTiter Glo assays. These results suggest a correlation among cell stiffness, cytoskeletal integrity, and susceptibility to recurrent exposures to nsPEFs, which emphasizes a mechanobiological underpinning of nsPEF bioeffects.
authors.affiliation: National Research Council, Joint Base San Antonio Fort Sam Houston, San Antonio, Texas.
authors.forename: Gary L
authors.initials: GL
authors.lastname: Thompson
authors.forename: Caleb
authors.initials: C
authors.lastname: Roth
authors.forename: Gleb
authors.initials: G
authors.lastname: Tolstykh
authors.forename: Marjorie
authors.initials: M
authors.lastname: Kuipers
authors.forename: Bennett L
authors.initials: BL
authors.lastname: Ibey
mesh_terms: D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000187:drug effects*; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D016466:CHO Cells; D002463:Cell Membrane Permeability / Q000187:drug effects; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D055119:Elastic Modulus / Q000187:drug effects; D004560:Electricity; D053778:Thiazolidines / Q000494:pharmacology
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; D053778:Thiazolidines; C037067:latrunculin A
keywords: elasticity; electroporation; electrosensitization; latrunculin; nanopore