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Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications
A fast re-endothelialization, along with the inhibition of neointima hyperplasia, are crucial to reduce the failure of vascular bypass grafts. Implants modifications with molecules capable of speeding up the re-endothelialization process have been proposed over the last years. However, clinical tria...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6465514/ https://www.ncbi.nlm.nih.gov/pubmed/31024906 http://dx.doi.org/10.3389/fbioe.2019.00074 |
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author | Copes, Francesco Chevallier, Pascale Loy, Caroline Pezzoli, Daniele Boccafoschi, Francesca Mantovani, Diego |
author_facet | Copes, Francesco Chevallier, Pascale Loy, Caroline Pezzoli, Daniele Boccafoschi, Francesca Mantovani, Diego |
author_sort | Copes, Francesco |
collection | PubMed |
description | A fast re-endothelialization, along with the inhibition of neointima hyperplasia, are crucial to reduce the failure of vascular bypass grafts. Implants modifications with molecules capable of speeding up the re-endothelialization process have been proposed over the last years. However, clinical trials of angiogenic factor delivery have been mostly disappointing, underscoring the need to investigate a wider array of angiogenic factors. In this work, a drug release system based on a type I collagen hydrogel has been proposed for the controlled release of Pleiotrophin (PTN), a cytokine known for its pro-angiogenetic effects. Heparin, in virtue of its ability to sequester, protect and release growth factors, has been used to better control the release of PTN. Performances of the PTN drug delivery system on endothelial (ECs) and smooth muscle cells (SMCs) have been investigated. Structural characterization (mechanical tests and immunofluorescent analyses of the collagen fibers) was performed on the gels to assess if heparin caused changes in their mechanical behavior. The release of PTN from the different gel formulations has been analyzed using a PTN-specific ELISA assay. Cell viability was evaluated with the Alamar Blue Cell Viability Assay on cells directly seeded on the gels (direct test) and on cells incubated with supernatant, containing the released PTN, obtained from the gels (indirect test). The effects of the different gels on the migration of both ECs and SMCs have been evaluated using a Transwell migration assay. Hemocompatibility of the gel has been assessed with a clotting/hemolysis test. Structural analyses showed that heparin did not change the structural behavior of the collagen gels. ELISA quantification demonstrated that heparin induced a constant release of PTN over time compared to other conditions. Both direct and indirect viability assays showed an increase in ECs viability while no effects were noted on SMCs. Cell migration results evidenced that the heparin/PTN-modified gels significantly increased ECs migration and decreased the SMCs one. Finally, heparin significantly increased the hemocompatibility of the collagen gels. In conclusion, the PTN-heparin-modified collagen here proposed can represent an added value for vascular medicine, able to ameliorate the biological performance, and integration of vascular grafts. |
format | Online Article Text |
id | pubmed-6465514 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-64655142019-04-25 Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications Copes, Francesco Chevallier, Pascale Loy, Caroline Pezzoli, Daniele Boccafoschi, Francesca Mantovani, Diego Front Bioeng Biotechnol Bioengineering and Biotechnology A fast re-endothelialization, along with the inhibition of neointima hyperplasia, are crucial to reduce the failure of vascular bypass grafts. Implants modifications with molecules capable of speeding up the re-endothelialization process have been proposed over the last years. However, clinical trials of angiogenic factor delivery have been mostly disappointing, underscoring the need to investigate a wider array of angiogenic factors. In this work, a drug release system based on a type I collagen hydrogel has been proposed for the controlled release of Pleiotrophin (PTN), a cytokine known for its pro-angiogenetic effects. Heparin, in virtue of its ability to sequester, protect and release growth factors, has been used to better control the release of PTN. Performances of the PTN drug delivery system on endothelial (ECs) and smooth muscle cells (SMCs) have been investigated. Structural characterization (mechanical tests and immunofluorescent analyses of the collagen fibers) was performed on the gels to assess if heparin caused changes in their mechanical behavior. The release of PTN from the different gel formulations has been analyzed using a PTN-specific ELISA assay. Cell viability was evaluated with the Alamar Blue Cell Viability Assay on cells directly seeded on the gels (direct test) and on cells incubated with supernatant, containing the released PTN, obtained from the gels (indirect test). The effects of the different gels on the migration of both ECs and SMCs have been evaluated using a Transwell migration assay. Hemocompatibility of the gel has been assessed with a clotting/hemolysis test. Structural analyses showed that heparin did not change the structural behavior of the collagen gels. ELISA quantification demonstrated that heparin induced a constant release of PTN over time compared to other conditions. Both direct and indirect viability assays showed an increase in ECs viability while no effects were noted on SMCs. Cell migration results evidenced that the heparin/PTN-modified gels significantly increased ECs migration and decreased the SMCs one. Finally, heparin significantly increased the hemocompatibility of the collagen gels. In conclusion, the PTN-heparin-modified collagen here proposed can represent an added value for vascular medicine, able to ameliorate the biological performance, and integration of vascular grafts. Frontiers Media S.A. 2019-04-09 /pmc/articles/PMC6465514/ /pubmed/31024906 http://dx.doi.org/10.3389/fbioe.2019.00074 Text en Copyright © 2019 Copes, Chevallier, Loy, Pezzoli, Boccafoschi and Mantovani. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Copes, Francesco Chevallier, Pascale Loy, Caroline Pezzoli, Daniele Boccafoschi, Francesca Mantovani, Diego Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications |
title | Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications |
title_full | Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications |
title_fullStr | Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications |
title_full_unstemmed | Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications |
title_short | Heparin-Modified Collagen Gels for Controlled Release of Pleiotrophin: Potential for Vascular Applications |
title_sort | heparin-modified collagen gels for controlled release of pleiotrophin: potential for vascular applications |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6465514/ https://www.ncbi.nlm.nih.gov/pubmed/31024906 http://dx.doi.org/10.3389/fbioe.2019.00074 |
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