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In vitro calcification studies on bioprosthetic and decellularized heart valves under quasi-physiological flow conditions

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Abstract

The lifespan of biological heart valve prostheses available in the market is limited due to structural alterations caused by calcium phosphate deposits formed from blood plasma in contact with the tissues. The objective of this work is to present a comparative methodology for the investigation of the formation of calcium phosphate deposits on bioprosthetic and tissue-engineered scaffolds in vitro and the influence of mechanical forces on tissue mineralization. Based on earlier investigations on biological mineralization at constant supersaturation, a circulatory loop simulating dynamic blood flow and physiological pressure conditions was developed. The system was appropriately adapted to evaluate the calcification potential of decellularized (DCV) and glutaraldehyde-fixed (GAV) porcine aortic valves. Results indicated that DCV calcified at higher, statistically nonsignificant, rates in comparison with GAV. This difference was attributed to the tissue surface modifications and cell debris leftovers from the decellularization process. Morphological analysis of the solids deposited after 20 h by scanning electron microscopy in combination with chemical microanalysis electron-dispersive spectroscopy identified the solid formed as octacalcium phosphate (Ca8(PO4)6H2·5H2O, OCP). OCP crystallites were preferentially deposited in high mechanical stress areas of the test tissues. Moreover, GAV tissues developed a significant transvalvular pressure gradient increase past 36 h with a calcium deposition distribution similar to the one found in explanted prostheses. In conclusion, the presented in vitro circulatory model serves as a valuable prescreening methodology for the investigation of the calcification process of bioprosthetic and tissue-engineered valves under physiological mechanical load.

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Acknowledgements

This research was funded by the People Program (Marie Curie Actions) of the European Union's Seventh Framework FP7/2007–2013/ under REA grant agreement n°317512.

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Authors and Affiliations

  1. Laboratory of Biomechanics and Biomedical Engineering, Department of Mechanical Engineering and Aeronautics, University of Patras, Patras, Greece

    Cristian C. D’Alessandro, Andreas Dimopoulos & Dimosthenis Mavrilas

  2. Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany

    Sofia Andriopoulou & Sotirios Korossis

  3. Department of Medical Physics, University Hospital of Patras, Patras, Greece

    Gerasimos A. T. Messaris

  4. Wolfson School of Mechanical, Electrical and Manufacturing Engineering, Loughborough University, Loughborough, UK

    Sotirios Korossis

  5. Department of Chemical Engineering, University of Patras, Patras, Greece

    Petros Koutsoukos

  6. FORTH-Institute of Chemical Engineering Sciences, Patras, Greece

    Petros Koutsoukos

Authors
  1. Cristian C. D’Alessandro
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  2. Andreas Dimopoulos
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  4. Gerasimos A. T. Messaris
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  5. Sotirios Korossis
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  7. Dimosthenis Mavrilas
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Contributions

CD contributed to data curation, formal analysis, investigation, methodology, software, validation, visualization and writing - original draft. AD helped in data curation, investigation and writing - review & editing. SA contributed to investigation. GM contributed to investigation. SK helped in conceptualization, funding acquisition and supervision. PK contributed to conceptualization, methodology, supervision, validation and writing - review & editing. DM helped in conceptualization, funding acquisition, methodology, supervision, resources, validation and writing - review & editing.

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Correspondence to Dimosthenis Mavrilas.

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D’Alessandro, C.C., Dimopoulos, A., Andriopoulou, S. et al. In vitro calcification studies on bioprosthetic and decellularized heart valves under quasi-physiological flow conditions. Bio-des. Manuf. 4, 10–21 (2021). https://doi.org/10.1007/s42242-020-00110-7

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