Synthetic biodegradable microporous hydrogels for in vitro 3D culture of functional human bone cell networks

Doris Zauchner, Monica Zippora Müller, Marion Horrer, Leana Bissig, Feihu Zhao, Philipp Fisch, Sung Sik Lee, Marcy Zenobi-Wong, Ralph Müller and Xiao-Hua Qin ()
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Doris Zauchner: ETH Zurich
Monica Zippora Müller: ETH Zurich
Marion Horrer: ETH Zurich
Leana Bissig: ETH Zurich
Feihu Zhao: Swansea University
Philipp Fisch: ETH Zurich
Sung Sik Lee: ETH Zurich
Marcy Zenobi-Wong: ETH Zurich
Ralph Müller: ETH Zurich
Xiao-Hua Qin: ETH Zurich

Nature Communications, 2024, vol. 15, issue 1, 1-12

Abstract: Abstract Generating 3D bone cell networks in vitro that mimic the dynamic process during early bone formation remains challenging. Here, we report a synthetic biodegradable microporous hydrogel for efficient formation of 3D networks from human primary cells, analysis of cell-secreted extracellular matrix (ECM) and microfluidic integration. Using polymerization-induced phase separation, we demonstrate dynamic in situ formation of microporosity (5–20 µm) within matrix metalloproteinase-degradable polyethylene glycol hydrogels in the presence of living cells. Pore formation is triggered by thiol-Michael-addition crosslinking of a viscous precursor solution supplemented with hyaluronic acid and dextran. The resulting microporous architecture can be fine-tuned by adjusting the concentration and molecular weight of dextran. After encapsulation in microporous hydrogels, human mesenchymal stromal cells and osteoblasts spread rapidly and form 3D networks within 24 hours. We demonstrate that matrix degradability controls cell-matrix remodeling, osteogenic differentiation, and deposition of ECM proteins such as collagen. Finally, we report microfluidic integration and proof-of-concept osteogenic differentiation of 3D cell networks under perfusion on chip. Altogether, this work introduces a synthetic microporous hydrogel to efficiently differentiate 3D human bone cell networks, facilitating future in vitro studies on early bone development.

Date: 2024
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DOI: 10.1038/s41467-024-49280-3

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