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Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model

SIMPLE SUMMARY: The chorioallantoic membrane (CAM) is an avian extraembryonic membrane widely used as an experimental assay to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the CAM has become popular in scientific studies focused on the use...

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Autores principales: Demcisakova, Zuzana, Luptakova, Lenka, Tirpakova, Zuzana, Kvasilova, Alena, Medvecky, Lubomir, De Spiegelaere, Ward, Petrovova, Eva
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9454696/
https://www.ncbi.nlm.nih.gov/pubmed/36077732
http://dx.doi.org/10.3390/cancers14174194
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author Demcisakova, Zuzana
Luptakova, Lenka
Tirpakova, Zuzana
Kvasilova, Alena
Medvecky, Lubomir
De Spiegelaere, Ward
Petrovova, Eva
author_facet Demcisakova, Zuzana
Luptakova, Lenka
Tirpakova, Zuzana
Kvasilova, Alena
Medvecky, Lubomir
De Spiegelaere, Ward
Petrovova, Eva
author_sort Demcisakova, Zuzana
collection PubMed
description SIMPLE SUMMARY: The chorioallantoic membrane (CAM) is an avian extraembryonic membrane widely used as an experimental assay to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the CAM has become popular in scientific studies focused on the use of its potential for the study of biocompatibility of materials for regenerative strategies and tissue engineering applications. Great research efforts are being made to develop innovative biomaterials able to treat hard tissue defects, including diseases such as a bone cancer. In this article, we describe an approach to detect the formation of blood vessels inside the porous acellular biopolymer polyhydroxybutyrate/chitosan (PHB/CHIT) scaffold using the CAM assay as an in vivo alternative animal model, including macroscopic, histological, immunohistochemical, and molecular evaluation of the biocompatibility. ABSTRACT: The chorioallantoic membrane (CAM) is a highly vascularized avian extraembryonic membrane widely used as an in vivo model to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the use of CAM has become an integral part of the biocompatibility testing process for developing biomaterials intended for regenerative strategies and tissue engineering applications. In this study, we used the chicken ex ovo CAM assay to investigate the angiogenic potential of innovative acellular biopolymer polyhydroxybutyrate/chitosan (PHB/CHIT) scaffold, which is intended for the treatment of hard tissue defects, depending on treatment with pro- and anti-angiogenic substances. On embryonic day (ED) 7, the experimental biomaterials were placed on the CAM alone or soaked in vascular endothelial growth factor (VEGF-A), saline solution (PHY), or tyrosine kinase inhibitor (SU5402). After 72 h, the formation of vessels was analyzed in the surrounding area of the scaffold and inside the pores of the implants, using markers of embryonic endothelium (WGA, SNA), myofibroblasts (α-SMA), and macrophages (KUL-01). The morphological and histochemical analysis showed strong angiogenic potential of untreated scaffolds without additional effect of the angiogenic factor, VEGF-A. The lowest angiogenic potential was observed in scaffolds soaked with SU5402. Gene expression of pro-angiogenic growth factors, i.e., VEGF-A, ANG-2, and VE-CAD, was upregulated in untreated scaffolds after 72 h, indicating a pro-angiogenic environment. We concluded that the PHB/CHIT has a strong endogenous angiogenic potential and could be promising biomaterial for the treatment of hard tissue defects.
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spelling pubmed-94546962022-09-09 Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model Demcisakova, Zuzana Luptakova, Lenka Tirpakova, Zuzana Kvasilova, Alena Medvecky, Lubomir De Spiegelaere, Ward Petrovova, Eva Cancers (Basel) Article SIMPLE SUMMARY: The chorioallantoic membrane (CAM) is an avian extraembryonic membrane widely used as an experimental assay to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the CAM has become popular in scientific studies focused on the use of its potential for the study of biocompatibility of materials for regenerative strategies and tissue engineering applications. Great research efforts are being made to develop innovative biomaterials able to treat hard tissue defects, including diseases such as a bone cancer. In this article, we describe an approach to detect the formation of blood vessels inside the porous acellular biopolymer polyhydroxybutyrate/chitosan (PHB/CHIT) scaffold using the CAM assay as an in vivo alternative animal model, including macroscopic, histological, immunohistochemical, and molecular evaluation of the biocompatibility. ABSTRACT: The chorioallantoic membrane (CAM) is a highly vascularized avian extraembryonic membrane widely used as an in vivo model to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the use of CAM has become an integral part of the biocompatibility testing process for developing biomaterials intended for regenerative strategies and tissue engineering applications. In this study, we used the chicken ex ovo CAM assay to investigate the angiogenic potential of innovative acellular biopolymer polyhydroxybutyrate/chitosan (PHB/CHIT) scaffold, which is intended for the treatment of hard tissue defects, depending on treatment with pro- and anti-angiogenic substances. On embryonic day (ED) 7, the experimental biomaterials were placed on the CAM alone or soaked in vascular endothelial growth factor (VEGF-A), saline solution (PHY), or tyrosine kinase inhibitor (SU5402). After 72 h, the formation of vessels was analyzed in the surrounding area of the scaffold and inside the pores of the implants, using markers of embryonic endothelium (WGA, SNA), myofibroblasts (α-SMA), and macrophages (KUL-01). The morphological and histochemical analysis showed strong angiogenic potential of untreated scaffolds without additional effect of the angiogenic factor, VEGF-A. The lowest angiogenic potential was observed in scaffolds soaked with SU5402. Gene expression of pro-angiogenic growth factors, i.e., VEGF-A, ANG-2, and VE-CAD, was upregulated in untreated scaffolds after 72 h, indicating a pro-angiogenic environment. We concluded that the PHB/CHIT has a strong endogenous angiogenic potential and could be promising biomaterial for the treatment of hard tissue defects. MDPI 2022-08-30 /pmc/articles/PMC9454696/ /pubmed/36077732 http://dx.doi.org/10.3390/cancers14174194 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Demcisakova, Zuzana
Luptakova, Lenka
Tirpakova, Zuzana
Kvasilova, Alena
Medvecky, Lubomir
De Spiegelaere, Ward
Petrovova, Eva
Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model
title Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model
title_full Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model
title_fullStr Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model
title_full_unstemmed Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model
title_short Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model
title_sort evaluation of angiogenesis in an acellular porous biomaterial based on polyhydroxybutyrate and chitosan using the chicken ex ovo chorioallantoic membrane model
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9454696/
https://www.ncbi.nlm.nih.gov/pubmed/36077732
http://dx.doi.org/10.3390/cancers14174194
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