TY - JOUR
T1 - Cytoskeletal Remodeling and Gap Junction Translocation Mediates Blood–Brain Barrier Disruption by Non-invasive Low-Voltage Pulsed Electric Fields
AU - Rajagopalan, Neeraj Raghuraman
AU - Vista, William-Ray
AU - Fujimori, Masashi
AU - Vroomen, Laurien G. P. H.
AU - Jiménez, Juan M.
AU - Khadka, Niranjan
AU - Bikson, Marom
AU - Srimathveeravalli, Govindarajan
N1 - Funding Information:
G.S. acknowledges grant and funding support from the National Cancer Institute and the National Institute of Diabetes, and Digestive and Kidney Diseases of the National Institutes of Health under Award Number U54CA137788/U54CA132378, R01CA236615 and R01DK129990, the Department of Defense CDMRP PRCRP Award CA170630, and the Institute for Applied Life Sciences in the University of Massachusetts at Amherst. MB is supported by Grants from Harold Shames and the National Institutes of Health: NIH-NIDA UG3DA048502, NIH-NIGMS T34 GM137858, NIH-NINDS R01 NS112996, NIH-NINDS R01 NS101362, and NIH-G-RISE T32GM136499.
Publisher Copyright:
© 2023, The Author(s) under exclusive licence to Biomedical Engineering Society.
PY - 2023
Y1 - 2023
N2 - High-voltage pulsed electric fields (HV-PEF) delivered with invasive needle electrodes for electroporation applications is known to induce off-target blood–brain barrier (BBB) disruption. In this study, we sought to determine the feasibility of minimally invasive PEF application to produce BBB disruption in rat brain and identify the putative mechanisms mediating the effect. We observed dose-dependent presence of Evans Blue (EB) dye in rat brain when PEF were delivered with a skull mounted electrode used for neurostimulation application. Maximum region of dye uptake was observed while using 1500 V, 100 pulses, 100 µs and 10 Hz. Results of computational models suggested that the region of BBB disruption was occurring at thresholds of 63 V/cm or higher; well below intensity levels for electroporation. In vitro experiments recapitulating this effect with human umbilical vein endothelial cells (HUVEC) demonstrated cellular alterations that underlie BBB manifests at low-voltage high-pulse conditions without affecting cell viability or proliferation. Morphological changes in HUVECs due to PEF were accompanied by disruption of actin cytoskeleton, loss of tight junction protein—ZO-1 and VE-Cadherin at cell junctions and partial translocation into the cytoplasm. Uptake of propidium iodide (PI) in PEF treated conditions is less than 1% and 2.5% of total number of cells in high voltage (HV) and low-voltage (LV) groups, respectively, implying that BBB disruption to be independent of electroporation under these conditions. 3-D microfabricated blood vessel permeability was found to increase significantly following PEF treatment and confirmed with correlative cytoskeletal changes and loss of tight junction proteins. Finally, we show that the rat brain model can be scaled to human brains with a similar effect on BBB disruption characterized by electric field strength (EFS) threshold and using a combination of two bilateral HD electrode configurations.
AB - High-voltage pulsed electric fields (HV-PEF) delivered with invasive needle electrodes for electroporation applications is known to induce off-target blood–brain barrier (BBB) disruption. In this study, we sought to determine the feasibility of minimally invasive PEF application to produce BBB disruption in rat brain and identify the putative mechanisms mediating the effect. We observed dose-dependent presence of Evans Blue (EB) dye in rat brain when PEF were delivered with a skull mounted electrode used for neurostimulation application. Maximum region of dye uptake was observed while using 1500 V, 100 pulses, 100 µs and 10 Hz. Results of computational models suggested that the region of BBB disruption was occurring at thresholds of 63 V/cm or higher; well below intensity levels for electroporation. In vitro experiments recapitulating this effect with human umbilical vein endothelial cells (HUVEC) demonstrated cellular alterations that underlie BBB manifests at low-voltage high-pulse conditions without affecting cell viability or proliferation. Morphological changes in HUVECs due to PEF were accompanied by disruption of actin cytoskeleton, loss of tight junction protein—ZO-1 and VE-Cadherin at cell junctions and partial translocation into the cytoplasm. Uptake of propidium iodide (PI) in PEF treated conditions is less than 1% and 2.5% of total number of cells in high voltage (HV) and low-voltage (LV) groups, respectively, implying that BBB disruption to be independent of electroporation under these conditions. 3-D microfabricated blood vessel permeability was found to increase significantly following PEF treatment and confirmed with correlative cytoskeletal changes and loss of tight junction proteins. Finally, we show that the rat brain model can be scaled to human brains with a similar effect on BBB disruption characterized by electric field strength (EFS) threshold and using a combination of two bilateral HD electrode configurations.
KW - Biological response to electroporation
KW - Blood–brain barrier (BBB) disruption
KW - Drug delivery
KW - Low-voltage pulsed electric field
UR - http://www.scopus.com/inward/record.url?scp=85153776209&partnerID=8YFLogxK
U2 - 10.1007/s10439-023-03211-3
DO - 10.1007/s10439-023-03211-3
M3 - Article
C2 - 37115366
SN - 0090-6964
JO - Annals of Biomedical Engineering
JF - Annals of Biomedical Engineering
ER -