Aspiration-assisted Bioprinting for Precise Positioning of Biologics

Introduction:: Cellular aggregates, such as spheroids, honeycombs, strands or even organoids, have been utilized in tissue fabrication. They have many advantages, including native-like cell density and the capability to secrete substantial levels of extracellular matrix (ECM) components with an effective communication among cells in a 3D native-like microenvironment. Cell aggregates, particularly spheroids, are excellent candidates as they can be assembled to form larger tissue complexes, such as but not limited to bone, pancreas and cardiac tissues. Spheroids loaded with endothelial cells can also facilitate a denser microenvironment, inherent ECM secretion, and pre vascular networks. These advantages make spheroids a strong candidate as building blocks for bioprinting of tissues.


Materials and Methods:: Despite the considerable interest in spheroids, their bioprinting have been rarely applied due to lack of robust, practical and scalable techniques. Only a few techniques, such as extrusion-based bioprinting, droplet-based bioprinting and Kenzan have been utilized; however, most of them suffer from poor positioning of spheroids, significant loss of their viability and structural integrity, poor process repeatability, and most importantly the lack of scalability. In our work [1], we introduced a technique, called ‘aspiration-assisted bioprinting,’ in which a single nozzle picks a spheroid by applying aspiration back-pressure and place into or onto a substrate.


Results, Conclusions, and Discussions:: Using aspiration-assisted bioprinting, spheroids with an order of magnitude size range (80-800 μm) were positioned successfully with minimal cellular damage ( >90% viability) and high positional precision (~11% with respect to spheroid size). We demonstrated the bioprinting of various types of spheroids both on gel substrates and within sacrificial or functional yield-stress support gels for fabrication of tissue such as bone [2-4], cartilage [4], osteochondral interfaces [5-6], cancer models [7-8], etc. We further advanced the technology to facilitate high-throughput positioning at an unprecedented speed with a high cell viability ( >90%).


Acknowledgements (Optional): : This work has been supported by NSF Award 1914885, NIH Award R01DE028614, R01EB034566, and 2236 CoCirculation2 of TUBITAK award # 121C359.


References (Optional): : < ![if !supportLists] >1. < ![endif] >Ayan, B., Heo, D., Zhang, Z., Dey, M., Pavilianskas, A., Drapaca, C., and Ozbolat, I.T., 2020, “Aspiration-assisted Bioprinting for Precise Positioning of Biologics” Science Advances, 6 (10), eaaw5111

< ![if !supportLists] >2. < ![endif] >Heo, D., Ayan, B., Dey, M., Banerjee, D., Lewis, G., Wee, H., and Ozbolat I.T., 2021, Aspiration-assisted bioprinting of co-cultured osteogenic spheroids for bone tissue fabrication, Biofabrication, 13(1), 015013

< ![if !supportLists] >3. < ![endif] >Kim, M.H., Banerjee, D., Celik, N., and Ozbolat, I.T., 2022 "Aspiration-assisted freeform bioprinting of mesenchymal stem cell spheroids within alginate microgels" Biofabrication, 14, 024103

< ![if !supportLists] >4. < ![endif] >Ayan, B, Celik, N., Zhang, Z., Zhou, K., Wu, Y., Costanzo, F., and Ozbolat, I.T., 2020, Aspiration-assisted Freeform Bioprinting of Pre-fabricated Tissue Spheroids in a Yield-stress Gel, Communications Physics, 3, 183

< ![if !supportLists] >5. < ![endif] >Wu, Y. , Ayan, B. , Vengadeshprabhu, F., Kamal, F., and Ozbolat, I.T., 2020, “Aspiration-assisted bioprinting of osteochondral interfaces,” Scientific Reports, 10, 13148.

< ![if !supportLists] >6. < ![endif] >Celik, N., Kim, M.Y., Yeo, M., Kamal, F., Hayes, D.J., Ozbolat, I.T., 2022 "miRNA Induced 3D Bioprinted-Heterotypic Osteochondral Interface" Biofabrication, 14(4), 044104.

< ![if !supportLists] >7. < ![endif] >Dey., M., Kim, M.H., Nagamine, M., Karhan, E., Kozhaya, L., Dogan, M., Unutmaz, D., Ozbolat, I.T., 2022 "Biofabrication of 3D breast cancer models for dissecting the cytotoxic response of human T cells expressing engineered MAIT cell receptors " Biofabrication, 14(4), 044105.

< ![if !supportLists] >8. < ![endif] >Dey, M., Kim, M.H., Dogan, M., Nagamine, M., Kozhaya, L., Celik, N., Unutmaz, D., Ozbolat, I.T., 2022, "Chemotherapeutics and CAR-T cell-based immunotherapeutics screening on a 3D bioprinted vascularized breast tumor model" Advanced Functional Materials, 202203966.