Perfusion-based co-culture model system for bone tissue engineering

Stephen W. Sawyer; Kairui Zhang; Jason A. Horton; Pranav Soman; Stephen W. Sawyer; Kairui Zhang; Jason A. Horton; Pranav Soman

doi:10.3934/bioeng.2020009

AIMS Bioengineering

2020, Volume 7, Issue 2: 91-105. doi: 10.3934/bioeng.2020009

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Perfusion-based co-culture model system for bone tissue engineering

1.
Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY, USA
2.
Department of Orthopedic Surgery, SUNY Upstate Medical University, Syracuse, NY, USA

Received: 07 May 2020 Accepted: 25 May 2020 Published: 29 May 2020

In this work, we report on a perfusion-based co-culture system that could be used for bone tissue engineering applications. The model system is created using a combination of Primary Human Umbilical Vein Endothelial Cells (HUVECs) and osteoblast-like Saos-2 cells encapsulated within a Gelatin Methacrylate (GelMA)-collagen hydrogel blend contained within 3D printed, perfusable constructs. The constructs contain dual channels, within a custom-built bioreactor, that were perfused with osteogenic media for up to two weeks in order to induce mineral deposition. Mineral deposition in constructs containing only HUVECs, only Saos-2 cells, or a combination thereof was quantified by microCT to determine if the combination of endothelial cells and bone-like cells increased mineral deposition. Histological and fluorescent staining was used to verify mineral deposition and cellular function both along and between the perfused channels. While there was not a quantifiable difference in the amount of mineral deposited in Saos-2 only versus Saos-2 plus HUVEC samples, the location of the deposited mineral differed dramatically between the groups and indicated that the addition of HUVECs within the GelMA matrix allowed Saos-2 cells, in diffusion limited regions of the construct, to deposit bone mineral. This work serves as a model on how to create perfusable bone tissue engineering constructs using a combination of 3D printing and cellular co-cultures.
- Gelatin methacrylate,
- mineral formation,
- cell encapsulation,
- perfusion,
- bioreactor,
- endothelial cells
Citation: Stephen W. Sawyer, Kairui Zhang, Jason A. Horton, Pranav Soman. Perfusion-based co-culture model system for bone tissue engineering[J]. AIMS Bioengineering, 2020, 7(2): 91-105. doi: 10.3934/bioeng.2020009

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Abstract

In this work, we report on a perfusion-based co-culture system that could be used for bone tissue engineering applications. The model system is created using a combination of Primary Human Umbilical Vein Endothelial Cells (HUVECs) and osteoblast-like Saos-2 cells encapsulated within a Gelatin Methacrylate (GelMA)-collagen hydrogel blend contained within 3D printed, perfusable constructs. The constructs contain dual channels, within a custom-built bioreactor, that were perfused with osteogenic media for up to two weeks in order to induce mineral deposition. Mineral deposition in constructs containing only HUVECs, only Saos-2 cells, or a combination thereof was quantified by microCT to determine if the combination of endothelial cells and bone-like cells increased mineral deposition. Histological and fluorescent staining was used to verify mineral deposition and cellular function both along and between the perfused channels. While there was not a quantifiable difference in the amount of mineral deposited in Saos-2 only versus Saos-2 plus HUVEC samples, the location of the deposited mineral differed dramatically between the groups and indicated that the addition of HUVECs within the GelMA matrix allowed Saos-2 cells, in diffusion limited regions of the construct, to deposit bone mineral. This work serves as a model on how to create perfusable bone tissue engineering constructs using a combination of 3D printing and cellular co-cultures.

Acknowledgments

This work was supported by (IGERT) DMR-DGE-1068780. This work was also partially supported by NIH R21GM129607 awarded to Pranav Soman. We would like to thank the Syracuse University Machine Shop, Lucas D. Albrecht, and Alex B. Filip for creating the polycarbonate bioreactors and printing the ABS cages. We would also like to thank Professor Megan E. Oest from SUNY Upstate Medical University for their aid in microCT analysis and histological sectioning, and Professor Zhen Ma for the use of his fluorescence microscope.

Conflict of interest

The authors declare no conflict of interest.

Author contribution

S.W.S., K.Z. J.A.H and P.S. wrote the manuscript; S.W.S. J.A.H and P.S. conceived and designed the experiments; S.W.S. and K.Z. performed cell studies and analyzed results.

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