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International Journal of Bioprinting Engineered EVs increase viability of 3D printed cardiomyocytes

cells , it is speculated that in the proposed system, the EVs fellowship from Kreitman School. Prof. Cohen holds the
[58]
mostly affect the bioink residing cells. However, further Claire and Harold Oshry Professor Chair in Biotechnology.
investigation of the effect of engineered EV released from
the patch is required to eliminate these concerns. Funding
The work was supported by the Jordan Baruch Stem Cell
4. Conclusion Fund: 87351721.
Herein, we described the successful engineering of a
macrophage-derived EV delivery system, capable of Conflict of interest
affecting multiple processes related to cardiac regeneration, The authors declare no conflict of interests.
including CM proliferation and cell death attenuation,
while also presenting angiogenic potential. The feasibility Author contributions
of miRNA cargo loading, while maintaining the EVs
membrane integrity, morphology and functionality, was Conceptualization: Assaf Bar, Smadar Cohen
demonstrated. These attributes have dramatic influence Formal analysis: Assaf Bar
upon the applicability of EV-based delivery systems. EV Investigation: Assaf Bar, Olga Kryukov
manipulation generally reduces EV yield [46,59] , affecting Methodology: Assaf Bar
treatment efficiency and cost. Moreover, EV stability Supervision: Smadar Cohen, Sharon Etzion
might affect their uptake and functionality upon delivery. Writing – original draft: Assaf Bar
The proposed EV delivery system exhibited capability to Writing – review & editing: Assaf Bar, Sharon Etzion,
harness new properties while maintaining their innate Smadar Cohen
beneficial traits; MΦ-EVs were also shown to affect ECs
in a way that could not only promote cell survival within Ethics approval and consent to participate
the CP, but also stimulate pre-vascularization if ECs are The study was carried out in strict accordance with the
incorporated within the patch. Guide for the Care and Use of Laboratory Animals of the
Currently, most efforts to improve 3D bioprinting National Institutes of Health. All animal studies reported
results are focused on changes in the bioink mechanical in this study were approved by the institutional ethics
properties and adjustments of printing parameters [60-62] . committee of Ben-Gurion University of the Negev, Israel
Here, the inclusion of the proposed engineered EVs within a (Protocol IL-72-09-2020(A)).
cell-laden bioink contributed to multiple aspects of cellular Consent for publication
viability, including CM survival, following 3D bioprinting.
Cell vitality is a major burden particularly in engineering Not applicable.
of CPs, where high concentrations of cells are required.
Proliferation of CM within CPs is considered to play a key Availability of data
role affecting the success of CP viability and integration Not applicable.
[63]
in vivo . To the best of our knowledge, this is the first
evidence on the successful induction of cardioprotection- References
related processes in CM post-3D bioprinting. The obtained
results could also be harnessed to improve the outcomes 1. Laflamme MA, Murry CE, 2011, Heart regeneration. Nature,
of 3D bioprinting for fabrication of more viable complex 473(7347):326.
cellular constructs. Furthermore, the engineered EV 2. Laflamme MA, Murry CE, 2005, Regenerating the heart. Nat
delivery system recapitulates multiple regenerative traits, Biotechnol, 23:845.
including angiogenesis. These could possibly provide the
CP with capability to integrate better with the host after 3. Liau B, Christoforou N, Leong KW, et al., 2011, Pluripotent
implementation in vivo. stem cell-derived cardiac tissue patch with advanced
structure and function. Biomaterials, 32(35):9180–9187.
Acknowledgments 4. Bar A, Cohen S, 2020, Inducing endogenous cardiac
regeneration: Can biomaterials connect the dots? Front
This work was done in partial fulfillment of the Bioeng Biotechnol, 8:126.
requirements for a PhD degree (NKAB) at the Avram 5. Tiburcy M, Hudson JE, Balfanz P, et al.2017, Defined
and Stella Goldstein-Goren Department of Biotechnology engineered human myocardium with advanced maturation
Engineering, Ben-Gurion University of the Negev, Israel. for applications in heart failure modeling and repair.
Assaf Bar gratefully acknowledges the BioTech Doctoral Circulation, 135(19):1832–1847.

Volume 9 Issue 2 (2023) 328 https://doi.org/10.18063/ijb.v9i2.670