File(s) under permanent embargo

Reason: This item is currently closed access.

Effects of combined cryopreservation and decellularization on the biomechanical, structural and biochemical properties of porcine pulmonary heart valves

journal contribution
posted on 01.03.2019, 13:45 by Karolina Theodoridis, Janina Muller, Robert Ramm, Katja Findeisen, Birgit Andree, Sotiris Korossis, Axel Haverich, Andres Hilfiker
Non-fixed, decellularized allogeneic heart valve scaffolds seem to be the best choice for heart valve replacement, their availability, however, is quite limited. Cryopreservation could prolong their shelf-life, allowing for their ideal match to a recipient. In this study, porcine pulmonary valves were decellularized using detergents, either prior or after cryopreservation, and analyzed. Mechanical integrity was analyzed by uniaxial tensile testing, histoarchitecture by histological staining, and composition by DNA, collagen (hydroxyproline) and GAG (chondroitin sulfate) quantification. Residual sodium dodecyl sulfate (SDS) in the scaffold was quantified by applying a methylene blue activation assay (MBAS). Cryopreserved decellularized scaffolds (DC) and scaffolds that were decellularized after cryopreservation (CD) were compared to fresh valves (F), cryopreserved native valves (C), and decellularized only scaffolds (D). The E-modulus and tensile strength of decellularized (D) tissue showed no significant difference compared to DC and CD. The decellularization resulted in an overall reduction of DNA and GAG, with DC containing the lowest amount of GAGs. The DNA content in the valvular wall of the CD group was higher than in the D and DC groups. CD valves showed slightly more residual SDS than DC valves, which might be harmful to recipient cells. In conclusion, cryopreservation after decellularization was shown to be preferable over cryopreservation before decellularization. However, in vivo testing would be necessary to determine whether these differences are significant in biocompatibility or immunogenicity of the scaffolds


This work was funded by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG) via the Cluster of Excellence ‘From regenerative biology to reconstructive therapy’ (REBIRTH) and via the C7 project of TRR127.



  • Mechanical, Electrical and Manufacturing Engineering

Published in

Acta Biomaterialia




71 - 77


THEODORIDIS, K. ... et al., 2016. Effects of combined cryopreservation and decellularization on the biomechanical, structural and biochemical properties of porcine pulmonary heart valves. Acta Biomaterialia, 43, pp. 71 - 77.


© Acta Materialia Inc. Published by Elsevier Ltd.


VoR (Version of Record)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at:

Publication date



This paper is in closed access.






Loughborough Publications