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. 2019 Oct 22;9:15117. doi: 10.1038/s41598-019-51513-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Bioactivity of released PDGF-BB from differently stored and UV sterilized bioactive DP scaffolds by Akt activation. (A) The phosphorylation status of Akt (pAkt) to dose-dependent PDGF-BB (0, 0.1, 0.5, 1, 10 and 50 ng/mL) supplementation (cells treated for 30 min) was analyzed by western blot. Presence of band for pAkt signifies phosphorylation of Akt and with that the ability of PDGF-BB to activate the PDGFR/PI3K/Akt pathway. The phosphorylation status of Akt (pAkt) to PDGF-BB released from (B) non-stored and non-sterilized DP scaffolds, (C) scaffolds stored for 1 week at room temperature (RT) or D) 4 °C or (E) −20 °C or (F) not stored but UV sterilized for 30 min. The release was performed in serum-free medium with 0.1% BSA and release samples for Day 1, 2 and 3 were tested (in duplicates), in order to obtain initial idea whether bioactivity of PDGF-BB is preserved after the specific treatments. Activation of Akt signaling pathway by released PDGF-BB, thus leading to increase in pAkt, shows retained bioactivity of the growth factor after the specific storage time period or the UV sterilization. For uncropped images, we refer to Fig. S1.