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Design of a Mechanobioreactor to Apply Anisotropic, Biaxial Strain to Large Thin Biomaterials for Tissue Engineered Heart Valve Applications

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Abstract

Repair and replacement solutions for congenitally diseased heart valves capable of post-surgery growth and adaptation have remained elusive. Tissue engineered heart valves (TEHVs) offer a potential biological solution that addresses the drawbacks of existing valve replacements. Typically, TEHVs are made from thin, fibrous biomaterials that either become cell populated in vitro or in situ. Often, TEHV designs poorly mimic the anisotropic mechanical properties of healthy native valves leading to inadequate biomechanical function. Mechanical conditioning of engineered tissues with anisotropic strain application can induce extracellular matrix remodelling to alter the anisotropic mechanical properties of a construct, but implementation has been limited to small-scale set-ups. To address this limitation for TEHV applications, we designed and built a mechanobioreactor capable of modulating biaxial strain anisotropy applied to large, thin, biomaterial sheets in vitro. The bioreactor can independently control two orthogonal stretch axes to modulate applied strain anisotropy on biomaterial sheets from 13 × 13 mm2 to 70 × 40 mm2. A design of experiments was performed using experimentally validated finite element (FE) models and demonstrated that biaxial strain was applied uniformly over a larger percentage of the cell seeded area for larger sheets (13 × 13 mm2: 58% of sheet area vs. 52 × 31 mm2: 86% of sheet area). Furthermore, bioreactor prototypes demonstrated that over 70% of the cell seeding area remained uniformly strained under different prescribed protocols: equibiaxial amplitudes between 5 to 40%, cyclic frequencies between 0.1 to 2.5 Hz and anisotropic strain ratios between 0:1 (constrained uniaxial) to 2:1. Lastly, proof-of-concept experiments were conducted where we applied equibiaxial (εx = εy = 8.75%) and anisotropic (εx = 12.5%, εy = 5%) strain protocols to cell-seeded, electrospun scaffolds. Cell nuclei and F-actin aligned to the vector-sum strain direction of each prescribed protocol (nuclei alignment: equibiaxial: 43.2° ± 1.8°, anisotropic: 17.5° ± 1.7°; p < 0.001). The abilities of this bioreactor to prescribe different strain amplitude, frequency and strain anisotropy protocols to cell-seeded scaffolds will enable future studies into the effects of anisotropic loading protocols on mechanically conditioned TEHVs and other engineered planar connective tissues.

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Acknowledgments

This work was supported by a Canadian Institute of Health Research/National Sciences and Engineering Research Council (NSERC) Collaborative Health Research Project (CPG-151962/ CHRPJ 508364-17) and seed funding from the Translational Biology and Engineering Program in the Ted Rogers Center for Heart Research. E.W was funded by an NSERC Canada Graduate Scholarships-Master’s award; an NSERC Postgraduate Scholarship-Doctorate award, an NSERC CONNECT! Collaborative Research & Training Experience program award; an Ontario Graduate Scholarship; and a Milligan Graduate Fellowship through the Department of Mechanical and Industrial Engineering at the University of Toronto. We would like to further acknowledge Dr. Cheryle A. Séguin and Dr. Mark M. K. Kim (Department of Physiology and Pharmacology, Western University, London ON) for their feedback on the bioreactor design.

Conflict of interest

All authors declare that they have no conflicts of interest and no financial ties related directly or indirectly to the subject of this manuscript.

Citation Diversity Statement

Recent studies have shown women and minority scholars have been under-cited relative to the proportion of publications in various scientific fields.2,5 Our group recognizes these biased citation practices are unacceptable. As such, we ensure our references are both appropriate given the scope of this work and is inclusive to all genders and races.

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  1. Translational Biology and Engineering Program, The Ted Rogers Centre for Heart Research, Toronto, Canada

    Edwin Wong, Shouka Parvin Nejad, Katya A. D’Costa, Nataly Machado Siqueira, Monica Lecce, J. Paul Santerre & Craig A. Simmons

  2. Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, Canada

    Edwin Wong & Craig A. Simmons

  3. Institute of Biomedical Engineering, University of Toronto, Toronto, Canada

    Edwin Wong, Shouka Parvin Nejad, Katya A. D’Costa, Nataly Machado Siqueira, Monica Lecce, J. Paul Santerre & Craig A. Simmons

  4. Faculty of Dentistry, University of Toronto, Toronto, Canada

    J. Paul Santerre

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Wong, E., Parvin Nejad, S., D’Costa, K.A. et al. Design of a Mechanobioreactor to Apply Anisotropic, Biaxial Strain to Large Thin Biomaterials for Tissue Engineered Heart Valve Applications. Ann Biomed Eng 50, 1073–1089 (2022). https://doi.org/10.1007/s10439-022-02984-3

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