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Abstract 3D bioprinting has been evolving as an important strategy for the fabrication of engineered tissues for clinical, diagnostic, and research applications. A major advantage of bioprinting is the ability to recapitulate the patient-specific tissue macro-architecture using cellular bioinks. The effectiveness of bioprinting can be significantly enhanced by incorporating the ability to preferentially organize cellular constituents within 3D constructs to mimic the intrinsic micro-architectural characteristics of native tissues. Accordingly, this work focuses on a new non-contact and label-free approach called ultrasound-assisted bioprinting (UAB) that utilizes acoustophoresis principle to align cells within bioprinted constructs. We describe the underlying process physics and develop and validate computational models to determine the effects of ultrasound process parameters (excitation mode, excitation time, frequency, voltage amplitude) on the relevant temperature, pressure distribution, and alignment time characteristics. Using knowledge from the computational models, we experimentally investigate the effect of selected process parameters (frequency, voltage amplitude) on the critical quality attributes (cellular strand width, inter-strand spacing, and viability) of MG63 cells in alginate as a model bioink system. Finally, we demonstrate the UAB of bilayered constructs with parallel (0°–0°) and orthogonal (0°–90°) cellular alignment across layers. Results of this work highlight the key interplay between the UAB process design and characteristics of aligned cellular constructs, and represent an important next step in our ability to create biomimetic engineered tissues.
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Ultrasonically-Induced Patterning of Viable Cells in Viscous Bioinks During 3D Biofabrication
In attempts to engineer human tissues in the lab, bio-mimicking the cellular arrangement of natural tissues is critical to achieve the required biological and mechanical form and function. Although biofabrication employing cellular bioinks continues to evolve as a promising solution over polymer scaffold based techniques in creating complex multi-cellular tissues, the ability of most current biofabrication processes to mimic the requisite cellular arrangement is limited. In this study, we propose a novel biofabrication approach that uses forces generated by bulk standing acoustic waves (BSAW) to non-deleteriously align cells within viscous bioinks. We computationally determine the acoustic pressure pattern generated by BSAW and experimentally map the effects of BSAW frequency (0.71, 1, 1.5, 2 MHz) on the linear arrangement of two types of human cells (adipose-derived stem cells and MG63) in alginate. Computational results indicate a non-linear relationship between frequency and acoustic pressure amplitude. Experimental results demonstrate that the spacing between adjacent strands of aligned cells is affected by frequency (p < 0.0001), and this effect is independent of the cell type. Lastly, we demonstrate a synergistic technique of gradual crosslinking in tandem with the BSAW-induced alignment to entrap cells within crosslinked hydrogels. This study represents an advancement in engineered tissue biofabrication aimed at bio-mimicry.
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- Award ID(s):
- 1652489
- PAR ID:
- 10157339
- Date Published:
- Journal Name:
- ASME 2019 14th International Manufacturing Science and Engineering Conference
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Conference Paper:
- https://doi.org/10.1115/MSEC2019-2816
