A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin

Autores
Saffioti, Nicolas Andres; Leal Denis, Maria Florencia; Alvarez, Cora Lilia; Herlax, Vanesa Silvana; Schwarzbaum, Pablo Julio; Pallarola, Diego Andres
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The α-haemolysin (HlyA) is an exotoxin produced by several strains of uropathogenic E. coli, one of the most important etiological agents of urinary infections. HlyA irreversibly binds to human erythrocytes (RBCs), initiating a degenerative process called eryptosis, characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the external layer of the plasma membrane of RBCs, shrinkage, and swelling. HlyA-induced PS externalization can lead to adhesion of RBCs to vascular endothelial cells (VECs). We studied the capacity of HlyA-treated RBCs to adhere to: 1- activated endothelial HMEC-1 cells under different flow conditions (dynamic adhesion); 2- surfaces homogeneously covered with extracellular matrix components in static conditions. Results showed that HlyA induced adhesion of RBCs to VECs at low flow (0.2 dyn/cm2), although higher flows induced rapid detachment. On the other hand, HlyA treatment also induced static adhesion of RBCs to collagen or fibrinogen. Thus, HlyA-treated RBCs displayed high but weak adherence to VECs under the experimental conditions. Additionally, to study the molecular mechanism of the HlyA-induced adhesion of RBCs we designed a biomimetic device to emulate the conditions of the blood vessels. The device was built by coupling a microfluidic chip to a nanopatterned surface (NPS) coated with gold nanoparticles (AuNPs). Different adhesion molecules from the VECs could be anchored to the AuNPs to mimic exposure of adhesion molecules of an activated endothelium whereas, the architecture of the capillaries was emulated by a network of microfluidic channels built in polydimethylsiloxane (PDMS). The synthesis of the device was optimized by following the process with quartz microbalance and fluorescence microscopy. Future experiments using the device will allow investigating HlyA-induced adhesion of RBCs to specific adhesion molecules under flow conditions.
Fil: Saffioti, Nicolas Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina
Fil: Leal Denis, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Alvarez, Cora Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Herlax, Vanesa Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina
Fil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Pallarola, Diego Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina
Primeras Jornadas Virtuales de la Sociedad Argentina de Biofísica
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Biofísica
Materia
MICROFLUIDICS
NANOPATTERNED
RTX TOXINS
ERIPTOSIS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/159529
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/159529
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysinSaffioti, Nicolas AndresLeal Denis, Maria FlorenciaAlvarez, Cora LiliaHerlax, Vanesa SilvanaSchwarzbaum, Pablo JulioPallarola, Diego AndresMICROFLUIDICSNANOPATTERNEDRTX TOXINSERIPTOSIShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The α-haemolysin (HlyA) is an exotoxin produced by several strains of uropathogenic E. coli, one of the most important etiological agents of urinary infections. HlyA irreversibly binds to human erythrocytes (RBCs), initiating a degenerative process called eryptosis, characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the external layer of the plasma membrane of RBCs, shrinkage, and swelling. HlyA-induced PS externalization can lead to adhesion of RBCs to vascular endothelial cells (VECs). We studied the capacity of HlyA-treated RBCs to adhere to: 1- activated endothelial HMEC-1 cells under different flow conditions (dynamic adhesion); 2- surfaces homogeneously covered with extracellular matrix components in static conditions. Results showed that HlyA induced adhesion of RBCs to VECs at low flow (0.2 dyn/cm2), although higher flows induced rapid detachment. On the other hand, HlyA treatment also induced static adhesion of RBCs to collagen or fibrinogen. Thus, HlyA-treated RBCs displayed high but weak adherence to VECs under the experimental conditions. Additionally, to study the molecular mechanism of the HlyA-induced adhesion of RBCs we designed a biomimetic device to emulate the conditions of the blood vessels. The device was built by coupling a microfluidic chip to a nanopatterned surface (NPS) coated with gold nanoparticles (AuNPs). Different adhesion molecules from the VECs could be anchored to the AuNPs to mimic exposure of adhesion molecules of an activated endothelium whereas, the architecture of the capillaries was emulated by a network of microfluidic channels built in polydimethylsiloxane (PDMS). The synthesis of the device was optimized by following the process with quartz microbalance and fluorescence microscopy. Future experiments using the device will allow investigating HlyA-induced adhesion of RBCs to specific adhesion molecules under flow conditions.Fil: Saffioti, Nicolas Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martin. Instituto de Nanosistemas; ArgentinaFil: Leal Denis, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Alvarez, Cora Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Herlax, Vanesa Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Pallarola, Diego Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martin. Instituto de Nanosistemas; ArgentinaPrimeras Jornadas Virtuales de la Sociedad Argentina de BiofísicaCiudad Autónoma de Buenos AiresArgentinaSociedad Argentina de BiofísicaSociedad Argentina de BiofíscaDelfino, Jose Maria2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectJornadaBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/159529A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin; Primeras Jornadas Virtuales de la Sociedad Argentina de Biofísica; Ciudad Autónoma de Buenos Aires; Argentina; 2020; 80-80978-987-27591-8-6CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:43:48Zoai:ri.conicet.gov.ar:11336/159529instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:43:48.415CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
title A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
spellingShingle A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
Saffioti, Nicolas Andres
MICROFLUIDICS
NANOPATTERNED
RTX TOXINS
ERIPTOSIS
title_short A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
title_full A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
title_fullStr A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
title_full_unstemmed A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
title_sort A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin
dc.creator.none.fl_str_mv Saffioti, Nicolas Andres
Leal Denis, Maria Florencia
Alvarez, Cora Lilia
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
Pallarola, Diego Andres
author Saffioti, Nicolas Andres
author_facet Saffioti, Nicolas Andres
Leal Denis, Maria Florencia
Alvarez, Cora Lilia
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
Pallarola, Diego Andres
author_role author
author2 Leal Denis, Maria Florencia
Alvarez, Cora Lilia
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
Pallarola, Diego Andres
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Delfino, Jose Maria
dc.subject.none.fl_str_mv MICROFLUIDICS
NANOPATTERNED
RTX TOXINS
ERIPTOSIS
topic MICROFLUIDICS
NANOPATTERNED
RTX TOXINS
ERIPTOSIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The α-haemolysin (HlyA) is an exotoxin produced by several strains of uropathogenic E. coli, one of the most important etiological agents of urinary infections. HlyA irreversibly binds to human erythrocytes (RBCs), initiating a degenerative process called eryptosis, characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the external layer of the plasma membrane of RBCs, shrinkage, and swelling. HlyA-induced PS externalization can lead to adhesion of RBCs to vascular endothelial cells (VECs). We studied the capacity of HlyA-treated RBCs to adhere to: 1- activated endothelial HMEC-1 cells under different flow conditions (dynamic adhesion); 2- surfaces homogeneously covered with extracellular matrix components in static conditions. Results showed that HlyA induced adhesion of RBCs to VECs at low flow (0.2 dyn/cm2), although higher flows induced rapid detachment. On the other hand, HlyA treatment also induced static adhesion of RBCs to collagen or fibrinogen. Thus, HlyA-treated RBCs displayed high but weak adherence to VECs under the experimental conditions. Additionally, to study the molecular mechanism of the HlyA-induced adhesion of RBCs we designed a biomimetic device to emulate the conditions of the blood vessels. The device was built by coupling a microfluidic chip to a nanopatterned surface (NPS) coated with gold nanoparticles (AuNPs). Different adhesion molecules from the VECs could be anchored to the AuNPs to mimic exposure of adhesion molecules of an activated endothelium whereas, the architecture of the capillaries was emulated by a network of microfluidic channels built in polydimethylsiloxane (PDMS). The synthesis of the device was optimized by following the process with quartz microbalance and fluorescence microscopy. Future experiments using the device will allow investigating HlyA-induced adhesion of RBCs to specific adhesion molecules under flow conditions.
Fil: Saffioti, Nicolas Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina
Fil: Leal Denis, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Alvarez, Cora Lilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Herlax, Vanesa Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentina
Fil: Schwarzbaum, Pablo Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Pallarola, Diego Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina
Primeras Jornadas Virtuales de la Sociedad Argentina de Biofísica
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Biofísica
description The α-haemolysin (HlyA) is an exotoxin produced by several strains of uropathogenic E. coli, one of the most important etiological agents of urinary infections. HlyA irreversibly binds to human erythrocytes (RBCs), initiating a degenerative process called eryptosis, characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the external layer of the plasma membrane of RBCs, shrinkage, and swelling. HlyA-induced PS externalization can lead to adhesion of RBCs to vascular endothelial cells (VECs). We studied the capacity of HlyA-treated RBCs to adhere to: 1- activated endothelial HMEC-1 cells under different flow conditions (dynamic adhesion); 2- surfaces homogeneously covered with extracellular matrix components in static conditions. Results showed that HlyA induced adhesion of RBCs to VECs at low flow (0.2 dyn/cm2), although higher flows induced rapid detachment. On the other hand, HlyA treatment also induced static adhesion of RBCs to collagen or fibrinogen. Thus, HlyA-treated RBCs displayed high but weak adherence to VECs under the experimental conditions. Additionally, to study the molecular mechanism of the HlyA-induced adhesion of RBCs we designed a biomimetic device to emulate the conditions of the blood vessels. The device was built by coupling a microfluidic chip to a nanopatterned surface (NPS) coated with gold nanoparticles (AuNPs). Different adhesion molecules from the VECs could be anchored to the AuNPs to mimic exposure of adhesion molecules of an activated endothelium whereas, the architecture of the capillaries was emulated by a network of microfluidic channels built in polydimethylsiloxane (PDMS). The synthesis of the device was optimized by following the process with quartz microbalance and fluorescence microscopy. Future experiments using the device will allow investigating HlyA-induced adhesion of RBCs to specific adhesion molecules under flow conditions.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Jornada
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/159529
A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin; Primeras Jornadas Virtuales de la Sociedad Argentina de Biofísica; Ciudad Autónoma de Buenos Aires; Argentina; 2020; 80-80
978-987-27591-8-6
CONICET Digital
CONICET
url http://hdl.handle.net/11336/159529
identifier_str_mv A biomimetic approach to study human erythrocytes adhesion to vascular endothelium induced by alpha-hemolysin; Primeras Jornadas Virtuales de la Sociedad Argentina de Biofísica; Ciudad Autónoma de Buenos Aires; Argentina; 2020; 80-80
978-987-27591-8-6
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Biofísca
publisher.none.fl_str_mv Sociedad Argentina de Biofísca
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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