Aspiration-assisted bioprinting of co-cultured osteogenic spheroids for bone tissue engineering

Dong Nyoung Heo; Bugra Ayan; Madhuri Dey; Dishary Banerjee; Hwabok Wee; Gregory S Lewis; Ibrahim T Ozbolat

doi:10.1088/1758-5090/abc1bf

Aspiration-assisted bioprinting of co-cultured osteogenic spheroids for bone tissue engineering

Biofabrication. 2020 Dec 17;13(1). doi: 10.1088/1758-5090/abc1bf.

Authors

Dong Nyoung Heo^{1

2

3}, Bugra Ayan^{1

2}, Madhuri Dey^{2

4}, Dishary Banerjee^{1

2}, Hwabok Wee⁵, Gregory S Lewis⁵, Ibrahim T Ozbolat^{1

2

6

7

8}

Affiliations

¹ Department of Engineering Science and Mechanics Department, Penn State University, University Park, PA 16802, United States of America.
² The Huck Institutes of the Life Sciences, Penn State University, University Park, PA 16802, United States of America.
³ Department of Dental Materials, School of Dentistry, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea.
⁴ Department of Chemistry, Penn State University, University Park, PA 16802, United States of America.
⁵ Department of Orthopaedics and Rehabilitation, Penn State College of Medicine, Hershey, PA 17033, United States of America.
⁶ Biomedical Engineering Department, Penn State University, University Park, PA 16802, United States of America.
⁷ Materials Research Institute, Penn State University, University Park, PA 16802, United States of America.
⁸ Department of Neurosurgery, Penn State College of Medicine, Hershey, PA 17033, United States of America.

PMID: 33059343
DOI: 10.1088/1758-5090/abc1bf

Abstract

Conventional top-down approaches in tissue engineering involving cell seeding on scaffolds have been widely used in bone engineering applications. However, scaffold-based bone tissue constructs have had limited clinical translation due to constrains in supporting scaffolds, minimal flexibility in tuning scaffold degradation, and low achievable cell seeding density as compared with native bone tissue. Here, we demonstrate a pragmatic and scalable bottom-up method, inspired from embryonic developmental biology, to build three-dimensional (3D) scaffold-free constructs using spheroids as building blocks. Human umbilical vein endothelial cells (HUVECs) were introduced to human mesenchymal stem cells (hMSCs) (hMSC/HUVEC) and spheroids were fabricated by an aggregate culture system. Bone tissue was generated by induction of osteogenic differentiation in hMSC/HUVEC spheroids for 10 d, with enhanced osteogenic differentiation and cell viability in the core of the spheroids compared to hMSC-only spheroids. Aspiration-assisted bioprinting (AAB) is a new bioprinting technique which allows precise positioning of spheroids (11% with respect to the spheroid diameter) by employing aspiration to lift individual spheroids and bioprint them onto a hydrogel. AAB facilitated bioprinting of scaffold-free bone tissue constructs using the pre-differentiated hMSC/HUVEC spheroids. These constructs demonstrated negligible changes in their shape for two days after bioprinting owing to the reduced proliferative potential of differentiated stem cells. Bioprinted bone tissues showed interconnectivity with actin-filament formation and high expression of osteogenic and endothelial-specific gene factors. This study thus presents a viable approach for 3D bioprinting of complex-shaped geometries using spheroids as building blocks, which can be used for various applications including but not limited to, tissue engineering, organ-on-a-chip and microfluidic devices, drug screening and, disease modeling.

Keywords: 3D bioprinting; aspiration-assisted bioprinting; biofabrication; bone tissue regeneration; osteogenic spheroids.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Bioprinting*
Bone and Bones
Human Umbilical Vein Endothelial Cells
Humans
Osteogenesis
Printing, Three-Dimensional
Spheroids, Cellular
Tissue Engineering
Tissue Scaffolds

Grants and funding

R01 DE028614/DE/NIDCR NIH HHS/United States