Designing of spider silk proteins for human induced pluripotent stem cell-based cardiac tissue engineering

T U Esser; V T Trossmann; S Lentz; F B Engel; T Scheibel

doi:10.1016/j.mtbio.2021.100114

Designing of spider silk proteins for human induced pluripotent stem cell-based cardiac tissue engineering

Mater Today Bio. 2021 May 15:11:100114. doi: 10.1016/j.mtbio.2021.100114. eCollection 2021 Jun.

Authors

T U Esser¹, V T Trossmann², S Lentz², F B Engel^{1

3}, T Scheibel^{2

4}

Affiliations

¹ Experimental Renal and Cardiovascular Research, Department of Nephropathology, Institute of Pathology, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Erlangen, 91054, Germany.
² Lehrstuhl Biomaterialien, Prof.-Rüdiger-Bormann Straße 1, Bayreuth, 95447, Germany.
³ MURCE, Muscle Research Center Erlangen, Erlangen, Germany.
⁴ Bayreuther Zentrum für Kolloide und Grenzflächen (BZKG), Bayerisches Polymerinstitut (BPI), Bayreuther Zentrum für Molekulare Biowissenschaften (BZMB), Bayreuther Materialzentrum (BayMAT), Universitätsstraße 30, Universität Bayreuth, Bayreuth, D-95447, Germany.

Abstract

Materials made of recombinant spider silk proteins are promising candidates for cardiac tissue engineering, and their suitability has so far been investigated utilizing primary rat cardiomyocytes. Herein, we expanded the tool box of available spider silk variants and demonstrated for the first time that human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes attach, contract, and respond to pharmacological treatment using phenylephrine and verapamil on explicit spider silk films. The hiPSC-cardiomyocytes contracted for at least 14 days on films made of positively charged engineered Araneus diadematus fibroin 4 (eADF4(κ16)) and three different arginyl-glycyl-aspartic acid (RGD)-tagged spider silk variants (positively or negatively charged and uncharged). Notably, hiPSC-cardiomyocytes exhibited different morphologies depending on the spider silk variant used, with less spreading and being smaller on films made of eADF4(κ16) than on RGD-tagged spider silk films. These results indicate that spider silk engineering is a powerful tool to provide new materials suitable for hiPSC-based cardiac tissue engineering.

Keywords: AFM, atomic force microscopy; APTES, (3-aminopropyl) triethoxysilane; ATR, attenuated total reflection; DPBS, Dulbecco's phosphate-buffered saline; EthHD1, ethidium homodimer 1; FT-IR, Fourier-transform infrared (spectroscopy); IPTG, isopropyl-β-D-thiogalactoside; MALDI-TOF, matrix-assisted laser desorption/ionization time-of-flight; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; eADF4, Engineered Araneus diadematus fibroin 4; hiPSC, human-induced pluripotent stem cell.