A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels

Autores
Saffioti, Nicolas Andres; Leal Denis, Maria Florencia; 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
Erythrocytes under pathological conditions undergo eryptosis, a process characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the plasma membrane external layer. Eryptotic erythrocytes adhere to the endothelial cells (ECs), which may be important in the pathology of bacterial infections and congenital diseases like sickle cell disease. Externalized PS can bind to receptors expressed on ECs under pathological conditions. To understand the adhesion mechanism, we designed a device combining microfluidics with nanostructured surfaces to mimic the capillary architecture and the expression of adhesive molecules by the activated ECs. Microfluidic chips were prepared in PDMS using molds fabricated by photolithography. Nanostructured surfaces were synthesized by block copolymer lithography and consisted of a glass surface covered with 7 nm diameter gold nanoparticles (AuNPs), arranged in a quasi-hexagonal array. The microfluidic chip was adhered to the nanostructured surface by an O2 plasma treatment. The AuNPs were functionalized with a polyethylene glycol chain (PEG) that binds to the AuNPs by a thiol at one end and has a nitriloacetic group (NTA) at the other end. The NTA binds proteins expressing a His-Tag. The surface not occupied by AuNPs was covered with PLL-g-PEG. We corroborated the specific AuNPs functionalization by quartz microbalance and fluorescence microscopy using a GFP-His Tag. Then, we studied the erythrocytes adhesion to a device functionalized with Annexin V-His Tag at different flows. Two conditions that promote eryptosis, incubation at 50° C or treatment with ionomycin, significantly increased the adhesion of erythrocytes at flows up to 1.5 dyn/cm2, in comparison with untreated erythrocytes. However, eryptotic erythrocytes also showed adherence to a device functionalized with a PEG lacking NTA groups. This indicates that erythrocytes adhesion may not be mediated only by PS receptors. Future experiments will test the erythrocytes adhesion elicited by proteins usually expressed by the activated ECs.
Fil: Saffioti, Nicolas Andres. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina. 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: 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: 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. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
20th Internaitional Congress of the INternational Union for Pure Applied Biophysics; 50th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology; 45th Congress of Brazilian Biophysics Society anda 13th Brazilian Society on Nuclear Biosciences Congress
Brasil
Sociedade Brasileira de Bioquímica e Biología Molecular
Materia
BIOMIMETIC
PHOSPHATIDYLSERINE
ENDOTHELIUM
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:11336/173664
Acceder
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vesselsSaffioti, Nicolas AndresLeal Denis, Maria FlorenciaHerlax, Vanesa SilvanaSchwarzbaum, Pablo JulioPallarola, Diego AndresBIOMIMETICPHOSPHATIDYLSERINEENDOTHELIUMhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Erythrocytes under pathological conditions undergo eryptosis, a process characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the plasma membrane external layer. Eryptotic erythrocytes adhere to the endothelial cells (ECs), which may be important in the pathology of bacterial infections and congenital diseases like sickle cell disease. Externalized PS can bind to receptors expressed on ECs under pathological conditions. To understand the adhesion mechanism, we designed a device combining microfluidics with nanostructured surfaces to mimic the capillary architecture and the expression of adhesive molecules by the activated ECs. Microfluidic chips were prepared in PDMS using molds fabricated by photolithography. Nanostructured surfaces were synthesized by block copolymer lithography and consisted of a glass surface covered with 7 nm diameter gold nanoparticles (AuNPs), arranged in a quasi-hexagonal array. The microfluidic chip was adhered to the nanostructured surface by an O2 plasma treatment. The AuNPs were functionalized with a polyethylene glycol chain (PEG) that binds to the AuNPs by a thiol at one end and has a nitriloacetic group (NTA) at the other end. The NTA binds proteins expressing a His-Tag. The surface not occupied by AuNPs was covered with PLL-g-PEG. We corroborated the specific AuNPs functionalization by quartz microbalance and fluorescence microscopy using a GFP-His Tag. Then, we studied the erythrocytes adhesion to a device functionalized with Annexin V-His Tag at different flows. Two conditions that promote eryptosis, incubation at 50° C or treatment with ionomycin, significantly increased the adhesion of erythrocytes at flows up to 1.5 dyn/cm2, in comparison with untreated erythrocytes. However, eryptotic erythrocytes also showed adherence to a device functionalized with a PEG lacking NTA groups. This indicates that erythrocytes adhesion may not be mediated only by PS receptors. Future experiments will test the erythrocytes adhesion elicited by proteins usually expressed by the activated ECs.Fil: Saffioti, Nicolas Andres. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina. 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: 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: 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. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina20th Internaitional Congress of the INternational Union for Pure Applied Biophysics; 50th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology; 45th Congress of Brazilian Biophysics Society anda 13th Brazilian Society on Nuclear Biosciences CongressBrasilSociedade Brasileira de Bioquímica e Biología MolecularSociedade Brasileira de Bioquímica e Biología Molecular2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/173664A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels; 20th Internaitional Congress of the INternational Union for Pure Applied Biophysics; 50th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology; 45th Congress of Brazilian Biophysics Society anda 13th Brazilian Society on Nuclear Biosciences Congress; Brasil; 2021; 1-2CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://easyapp.ekmf.com.br/sg/uploads/eventos/evento8/documentos/abstract_book.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:01:42Zoai:ri.conicet.gov.ar:11336/173664instacron: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:34982025-09-29 10:01:42.271CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
title A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
spellingShingle A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
Saffioti, Nicolas Andres
BIOMIMETIC
PHOSPHATIDYLSERINE
ENDOTHELIUM
title_short A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
title_full A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
title_fullStr A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
title_full_unstemmed A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
title_sort A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels
dc.creator.none.fl_str_mv Saffioti, Nicolas Andres
Leal Denis, Maria Florencia
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
Pallarola, Diego Andres
author Saffioti, Nicolas Andres
author_facet Saffioti, Nicolas Andres
Leal Denis, Maria Florencia
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
Pallarola, Diego Andres
author_role author
author2 Leal Denis, Maria Florencia
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
Pallarola, Diego Andres
author2_role author
author
author
author
dc.subject.none.fl_str_mv BIOMIMETIC
PHOSPHATIDYLSERINE
ENDOTHELIUM
topic BIOMIMETIC
PHOSPHATIDYLSERINE
ENDOTHELIUM
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Erythrocytes under pathological conditions undergo eryptosis, a process characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the plasma membrane external layer. Eryptotic erythrocytes adhere to the endothelial cells (ECs), which may be important in the pathology of bacterial infections and congenital diseases like sickle cell disease. Externalized PS can bind to receptors expressed on ECs under pathological conditions. To understand the adhesion mechanism, we designed a device combining microfluidics with nanostructured surfaces to mimic the capillary architecture and the expression of adhesive molecules by the activated ECs. Microfluidic chips were prepared in PDMS using molds fabricated by photolithography. Nanostructured surfaces were synthesized by block copolymer lithography and consisted of a glass surface covered with 7 nm diameter gold nanoparticles (AuNPs), arranged in a quasi-hexagonal array. The microfluidic chip was adhered to the nanostructured surface by an O2 plasma treatment. The AuNPs were functionalized with a polyethylene glycol chain (PEG) that binds to the AuNPs by a thiol at one end and has a nitriloacetic group (NTA) at the other end. The NTA binds proteins expressing a His-Tag. The surface not occupied by AuNPs was covered with PLL-g-PEG. We corroborated the specific AuNPs functionalization by quartz microbalance and fluorescence microscopy using a GFP-His Tag. Then, we studied the erythrocytes adhesion to a device functionalized with Annexin V-His Tag at different flows. Two conditions that promote eryptosis, incubation at 50° C or treatment with ionomycin, significantly increased the adhesion of erythrocytes at flows up to 1.5 dyn/cm2, in comparison with untreated erythrocytes. However, eryptotic erythrocytes also showed adherence to a device functionalized with a PEG lacking NTA groups. This indicates that erythrocytes adhesion may not be mediated only by PS receptors. Future experiments will test the erythrocytes adhesion elicited by proteins usually expressed by the activated ECs.
Fil: Saffioti, Nicolas Andres. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina. 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: 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: 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. Universidad Nacional de San Martin. Instituto de Nanosistemas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
20th Internaitional Congress of the INternational Union for Pure Applied Biophysics; 50th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology; 45th Congress of Brazilian Biophysics Society anda 13th Brazilian Society on Nuclear Biosciences Congress
Brasil
Sociedade Brasileira de Bioquímica e Biología Molecular
description Erythrocytes under pathological conditions undergo eryptosis, a process characterized by biochemical and morphological changes such as phosphatidylserine (PS) exposure to the plasma membrane external layer. Eryptotic erythrocytes adhere to the endothelial cells (ECs), which may be important in the pathology of bacterial infections and congenital diseases like sickle cell disease. Externalized PS can bind to receptors expressed on ECs under pathological conditions. To understand the adhesion mechanism, we designed a device combining microfluidics with nanostructured surfaces to mimic the capillary architecture and the expression of adhesive molecules by the activated ECs. Microfluidic chips were prepared in PDMS using molds fabricated by photolithography. Nanostructured surfaces were synthesized by block copolymer lithography and consisted of a glass surface covered with 7 nm diameter gold nanoparticles (AuNPs), arranged in a quasi-hexagonal array. The microfluidic chip was adhered to the nanostructured surface by an O2 plasma treatment. The AuNPs were functionalized with a polyethylene glycol chain (PEG) that binds to the AuNPs by a thiol at one end and has a nitriloacetic group (NTA) at the other end. The NTA binds proteins expressing a His-Tag. The surface not occupied by AuNPs was covered with PLL-g-PEG. We corroborated the specific AuNPs functionalization by quartz microbalance and fluorescence microscopy using a GFP-His Tag. Then, we studied the erythrocytes adhesion to a device functionalized with Annexin V-His Tag at different flows. Two conditions that promote eryptosis, incubation at 50° C or treatment with ionomycin, significantly increased the adhesion of erythrocytes at flows up to 1.5 dyn/cm2, in comparison with untreated erythrocytes. However, eryptotic erythrocytes also showed adherence to a device functionalized with a PEG lacking NTA groups. This indicates that erythrocytes adhesion may not be mediated only by PS receptors. Future experiments will test the erythrocytes adhesion elicited by proteins usually expressed by the activated ECs.
publishDate 2021
dc.date.none.fl_str_mv 2021
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info:eu-repo/semantics/conferenceObject
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status_str publishedVersion
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dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/173664
A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels; 20th Internaitional Congress of the INternational Union for Pure Applied Biophysics; 50th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology; 45th Congress of Brazilian Biophysics Society anda 13th Brazilian Society on Nuclear Biosciences Congress; Brasil; 2021; 1-2
CONICET Digital
CONICET
url http://hdl.handle.net/11336/173664
identifier_str_mv A biomimetic device combining microfluidics with nanotechnology allows studying the adhesion of erythrocytes to blood vessels; 20th Internaitional Congress of the INternational Union for Pure Applied Biophysics; 50th Annual Meeting of the Brazilian Society for Biochemistry and Molecular Biology; 45th Congress of Brazilian Biophysics Society anda 13th Brazilian Society on Nuclear Biosciences Congress; Brasil; 2021; 1-2
CONICET Digital
CONICET
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dc.publisher.none.fl_str_mv Sociedade Brasileira de Bioquímica e Biología Molecular
publisher.none.fl_str_mv Sociedade Brasileira de Bioquímica e Biología Molecular
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