Cargando…
ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan
Chitosan nanogel-coated polycaprolactone (PCL) fiber mat-based implant prototypes with tailored release of bone morphogenic protein 2 (BMP-2) are a promising approach to achieve implant-mediated bone regeneration. In order to ensure reliable in vitro release results, the robustness of a commercially...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073737/ https://www.ncbi.nlm.nih.gov/pubmed/33921903 http://dx.doi.org/10.3390/pharmaceutics13040582 |
_version_ | 1783684199021019136 |
---|---|
author | Sundermann, Julius Sydow, Steffen Burmeister, Laura Hoffmann, Andrea Menzel, Henning Bunjes, Heike |
author_facet | Sundermann, Julius Sydow, Steffen Burmeister, Laura Hoffmann, Andrea Menzel, Henning Bunjes, Heike |
author_sort | Sundermann, Julius |
collection | PubMed |
description | Chitosan nanogel-coated polycaprolactone (PCL) fiber mat-based implant prototypes with tailored release of bone morphogenic protein 2 (BMP-2) are a promising approach to achieve implant-mediated bone regeneration. In order to ensure reliable in vitro release results, the robustness of a commercially available ELISA for E. coli-derived BMP-2 and the parallel determination of BMP-2 recovery using a quantitative biological activity assay were investigated within a common release setup, with special reference to solubility and matrix effects. Without bovine serum albumin and Tween 20 as solubilizing additives to release media buffed at physiological pH, BMP-2 recoveries after release were notably reduced. In contrast, the addition of chitosan to release samples caused an excessive recovery. A possible explanation for these effects is the reversible aggregation tendency of BMP-2, which might be influenced by an interaction with chitosan. The interfering effects highlighted in this study are of great importance for bio-assay-based BMP-2 quantification, especially in the context of pharmaceutical release experiments. |
format | Online Article Text |
id | pubmed-8073737 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-80737372021-04-27 ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan Sundermann, Julius Sydow, Steffen Burmeister, Laura Hoffmann, Andrea Menzel, Henning Bunjes, Heike Pharmaceutics Article Chitosan nanogel-coated polycaprolactone (PCL) fiber mat-based implant prototypes with tailored release of bone morphogenic protein 2 (BMP-2) are a promising approach to achieve implant-mediated bone regeneration. In order to ensure reliable in vitro release results, the robustness of a commercially available ELISA for E. coli-derived BMP-2 and the parallel determination of BMP-2 recovery using a quantitative biological activity assay were investigated within a common release setup, with special reference to solubility and matrix effects. Without bovine serum albumin and Tween 20 as solubilizing additives to release media buffed at physiological pH, BMP-2 recoveries after release were notably reduced. In contrast, the addition of chitosan to release samples caused an excessive recovery. A possible explanation for these effects is the reversible aggregation tendency of BMP-2, which might be influenced by an interaction with chitosan. The interfering effects highlighted in this study are of great importance for bio-assay-based BMP-2 quantification, especially in the context of pharmaceutical release experiments. MDPI 2021-04-19 /pmc/articles/PMC8073737/ /pubmed/33921903 http://dx.doi.org/10.3390/pharmaceutics13040582 Text en © 2021 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 Sundermann, Julius Sydow, Steffen Burmeister, Laura Hoffmann, Andrea Menzel, Henning Bunjes, Heike ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan |
title | ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan |
title_full | ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan |
title_fullStr | ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan |
title_full_unstemmed | ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan |
title_short | ELISA- and Activity Assay-Based Quantification of BMP-2 Released In Vitro Can Be Biased by Solubility in “Physiological” Buffers and an Interfering Effect of Chitosan |
title_sort | elisa- and activity assay-based quantification of bmp-2 released in vitro can be biased by solubility in “physiological” buffers and an interfering effect of chitosan |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8073737/ https://www.ncbi.nlm.nih.gov/pubmed/33921903 http://dx.doi.org/10.3390/pharmaceutics13040582 |
work_keys_str_mv | AT sundermannjulius elisaandactivityassaybasedquantificationofbmp2releasedinvitrocanbebiasedbysolubilityinphysiologicalbuffersandaninterferingeffectofchitosan AT sydowsteffen elisaandactivityassaybasedquantificationofbmp2releasedinvitrocanbebiasedbysolubilityinphysiologicalbuffersandaninterferingeffectofchitosan AT burmeisterlaura elisaandactivityassaybasedquantificationofbmp2releasedinvitrocanbebiasedbysolubilityinphysiologicalbuffersandaninterferingeffectofchitosan AT hoffmannandrea elisaandactivityassaybasedquantificationofbmp2releasedinvitrocanbebiasedbysolubilityinphysiologicalbuffersandaninterferingeffectofchitosan AT menzelhenning elisaandactivityassaybasedquantificationofbmp2releasedinvitrocanbebiasedbysolubilityinphysiologicalbuffersandaninterferingeffectofchitosan AT bunjesheike elisaandactivityassaybasedquantificationofbmp2releasedinvitrocanbebiasedbysolubilityinphysiologicalbuffersandaninterferingeffectofchitosan |