ELISA- and Activity Assay-Based Quantification of BMP-2 Released <i>In Vitro</i> 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 <i>in vitro</i> release results, the robustnes...

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Bibliographic Details
Main Authors: Julius Sundermann, Steffen Sydow, Laura Burmeister, Andrea Hoffmann, Henning Menzel, Heike Bunjes
Format: Article
Language:English
Published: MDPI AG 2021-04-01
Series:Pharmaceutics
Subjects:
Online Access:https://www.mdpi.com/1999-4923/13/4/582
Description
Summary: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 <i>in vitro</i> release results, the robustness of a commercially available ELISA for <i>E. coli</i>-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.
ISSN:1999-4923