Evidence for injurious effect of cocaethylene in human microvascular endothelial cells

Danyel Hermes Tacker, Anthony Okorodudu

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: Cocaethylene (CE) is a conjugate of cocaine and ethanol that may contribute to the pathogenesis of systemic vascular diseases. This study was conducted to investigate the effect of CE on human microvascular endothelial cells (HMEC-1) in culture. Methods: Proliferating and confluent monolayers of HMEC-1 were used for assessing growth kinetics, viability, cytotoxicity, and morphologic/barrier alterations after CE treatment (0-1 mmol/l) for up to 7 days. The Trypan blue exclusion, lactate dehydrogenase (LDH) release assay, manual cell counts, and silver nitrate staining technique were used. Results: The doubling times of 30.0 and 31.4 h for the 0.5 and 1.0 mmol/l CE-treated HMEC-1, respectively, were significantly longer than the 28.6 h for the control group (p<0.05). The viabilities of 90.4±3.8% (control) and 93.1±1.9% (CE-treated) from the Trypan blue exclusion-staining experiments indicated non-lethality of CE. LDH activities of 173±33 U/l (control) and 157±43 U/l (CE-treated) confirmed the absence of CE cytotoxicity. Silver staining results indicated increased monolayer permeability as demonstrated by the formation of intercellular gaps after 1 h of exposure. Conclusions: HMEC-1 exposure to CE induced cellular injury that could affect the permeability of small blood vessels. These cellular changes could in part be the pivotal point for studies to explain the edema and inflammation in surrounding tissues of individuals exposed to CE.

Original languageEnglish (US)
Pages (from-to)69-77
Number of pages9
JournalClinica Chimica Acta
Volume345
Issue number1-2
DOIs
StatePublished - Jul 2004

Fingerprint

Endothelial cells
Endothelial Cells
Silver Staining
Trypan Blue
Cytotoxicity
L-Lactate Dehydrogenase
Monolayers
Permeability
cocaethylene
Silver Nitrate
Growth kinetics
Blood vessels
Vascular Diseases
Cocaine
Silver
Cell culture
Blood Vessels
Assays
Edema
Ethanol

Keywords

  • Cocaethylene
  • Cocaine
  • Endothelial cells
  • Ethanol
  • Microvascular

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Evidence for injurious effect of cocaethylene in human microvascular endothelial cells. / Tacker, Danyel Hermes; Okorodudu, Anthony.

In: Clinica Chimica Acta, Vol. 345, No. 1-2, 07.2004, p. 69-77.

Research output: Contribution to journalArticle

@article{0b9a7054c04848528b4751cd78679eaa,
title = "Evidence for injurious effect of cocaethylene in human microvascular endothelial cells",
abstract = "Background: Cocaethylene (CE) is a conjugate of cocaine and ethanol that may contribute to the pathogenesis of systemic vascular diseases. This study was conducted to investigate the effect of CE on human microvascular endothelial cells (HMEC-1) in culture. Methods: Proliferating and confluent monolayers of HMEC-1 were used for assessing growth kinetics, viability, cytotoxicity, and morphologic/barrier alterations after CE treatment (0-1 mmol/l) for up to 7 days. The Trypan blue exclusion, lactate dehydrogenase (LDH) release assay, manual cell counts, and silver nitrate staining technique were used. Results: The doubling times of 30.0 and 31.4 h for the 0.5 and 1.0 mmol/l CE-treated HMEC-1, respectively, were significantly longer than the 28.6 h for the control group (p<0.05). The viabilities of 90.4±3.8{\%} (control) and 93.1±1.9{\%} (CE-treated) from the Trypan blue exclusion-staining experiments indicated non-lethality of CE. LDH activities of 173±33 U/l (control) and 157±43 U/l (CE-treated) confirmed the absence of CE cytotoxicity. Silver staining results indicated increased monolayer permeability as demonstrated by the formation of intercellular gaps after 1 h of exposure. Conclusions: HMEC-1 exposure to CE induced cellular injury that could affect the permeability of small blood vessels. These cellular changes could in part be the pivotal point for studies to explain the edema and inflammation in surrounding tissues of individuals exposed to CE.",
keywords = "Cocaethylene, Cocaine, Endothelial cells, Ethanol, Microvascular",
author = "Tacker, {Danyel Hermes} and Anthony Okorodudu",
year = "2004",
month = "7",
doi = "10.1016/j.cccn.2004.02.031",
language = "English (US)",
volume = "345",
pages = "69--77",
journal = "Clinica Chimica Acta",
issn = "0009-8981",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Evidence for injurious effect of cocaethylene in human microvascular endothelial cells

AU - Tacker, Danyel Hermes

AU - Okorodudu, Anthony

PY - 2004/7

Y1 - 2004/7

N2 - Background: Cocaethylene (CE) is a conjugate of cocaine and ethanol that may contribute to the pathogenesis of systemic vascular diseases. This study was conducted to investigate the effect of CE on human microvascular endothelial cells (HMEC-1) in culture. Methods: Proliferating and confluent monolayers of HMEC-1 were used for assessing growth kinetics, viability, cytotoxicity, and morphologic/barrier alterations after CE treatment (0-1 mmol/l) for up to 7 days. The Trypan blue exclusion, lactate dehydrogenase (LDH) release assay, manual cell counts, and silver nitrate staining technique were used. Results: The doubling times of 30.0 and 31.4 h for the 0.5 and 1.0 mmol/l CE-treated HMEC-1, respectively, were significantly longer than the 28.6 h for the control group (p<0.05). The viabilities of 90.4±3.8% (control) and 93.1±1.9% (CE-treated) from the Trypan blue exclusion-staining experiments indicated non-lethality of CE. LDH activities of 173±33 U/l (control) and 157±43 U/l (CE-treated) confirmed the absence of CE cytotoxicity. Silver staining results indicated increased monolayer permeability as demonstrated by the formation of intercellular gaps after 1 h of exposure. Conclusions: HMEC-1 exposure to CE induced cellular injury that could affect the permeability of small blood vessels. These cellular changes could in part be the pivotal point for studies to explain the edema and inflammation in surrounding tissues of individuals exposed to CE.

AB - Background: Cocaethylene (CE) is a conjugate of cocaine and ethanol that may contribute to the pathogenesis of systemic vascular diseases. This study was conducted to investigate the effect of CE on human microvascular endothelial cells (HMEC-1) in culture. Methods: Proliferating and confluent monolayers of HMEC-1 were used for assessing growth kinetics, viability, cytotoxicity, and morphologic/barrier alterations after CE treatment (0-1 mmol/l) for up to 7 days. The Trypan blue exclusion, lactate dehydrogenase (LDH) release assay, manual cell counts, and silver nitrate staining technique were used. Results: The doubling times of 30.0 and 31.4 h for the 0.5 and 1.0 mmol/l CE-treated HMEC-1, respectively, were significantly longer than the 28.6 h for the control group (p<0.05). The viabilities of 90.4±3.8% (control) and 93.1±1.9% (CE-treated) from the Trypan blue exclusion-staining experiments indicated non-lethality of CE. LDH activities of 173±33 U/l (control) and 157±43 U/l (CE-treated) confirmed the absence of CE cytotoxicity. Silver staining results indicated increased monolayer permeability as demonstrated by the formation of intercellular gaps after 1 h of exposure. Conclusions: HMEC-1 exposure to CE induced cellular injury that could affect the permeability of small blood vessels. These cellular changes could in part be the pivotal point for studies to explain the edema and inflammation in surrounding tissues of individuals exposed to CE.

KW - Cocaethylene

KW - Cocaine

KW - Endothelial cells

KW - Ethanol

KW - Microvascular

UR - http://www.scopus.com/inward/record.url?scp=2942582367&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2942582367&partnerID=8YFLogxK

U2 - 10.1016/j.cccn.2004.02.031

DO - 10.1016/j.cccn.2004.02.031

M3 - Article

VL - 345

SP - 69

EP - 77

JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

SN - 0009-8981

IS - 1-2

ER -