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Affordable Oxygen Microscopy-Assisted Biofabrication of Multicellular Spheroids

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Oxygen mapping: Probing a novel seeding strategy for bone tissue engineering.

Ines Westphal^1,2, Claudia Jedelhauser¹, Gregor Liebsch³

¹Experimental Surgery and Regenerative Medicine, Department of General, Trauma and Reconstruction Surgery, University Hospital, Ludwig-Maximillians-University of Munich, Nussbaumstr. 20, Munich 80336, Germany.

Biotechnology and Bioengineering

|October 18, 2016

Summary

This summary is machine-generated.

Optimizing bone tissue engineering (BTE) requires careful control of human mesenchymal stem cell (hMSC) density. Harvesting hMSCs at lower densities for seeding biomaterial scaffolds improves engineered tissue outcomes and allows for earlier implantation.

Keywords:

3D scaffolds bone tissue engineering cell density mesenchymal stem cells oxygen measurement time point of harvest

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Area of Science:

Biomaterials Science
Regenerative Medicine
Cell Biology

Background:

Bone tissue engineering (BTE) aims to repair bone defects using biomaterial scaffolds and human mesenchymal stem cells (hMSCs).
Tissue quality in BTE is significantly influenced by cell density and oxygen availability within scaffolds.
Optimizing cell seeding and culture conditions is critical for successful BTE outcomes.

Purpose of the Study:

To introduce a novel oxygen-imaging sensor for monitoring oxygen distribution in 3D scaffolds.
To analyze a new cell-seeding strategy for demineralized bone matrix (DBM) scaffolds using hMSCs.
To investigate the impact of hMSC harvest density on oxygen consumption and tissue engineering efficacy.

Main Methods:

Development and application of a novel planar oxygen-imaging sensor for real-time oxygen mapping.
Simultaneous oxygen consumption measurements using the novel sensor and a conventional needle-type sensor over 24 hours.
Seeding DBM scaffolds with immortalized hMSCs pre-cultured to either 30-40% or 70-80% confluence.

Main Results:

Scaffolds seeded with hMSCs harvested at lower densities (30-40% confluence) showed rapid, exponential oxygen consumption.
Scaffolds seeded with hMSCs harvested at higher densities (70-80% confluence) exhibited slow, near-linear oxygen decrease, indicating a stationary growth phase.
The novel oxygen sensor provided continuous, 2D oxygen mapping, revealing distinct consumption patterns based on cell harvest density.

Conclusions:

Both scaffold seeding density and the density of harvested cells are critical parameters for successful BTE.
Harvesting hMSCs at lower densities during their log phase offers a promising strategy for earlier in vivo implantation of cell-seeded scaffolds.
The novel oxygen imaging sensor is a valuable tool for continuous, convenient oxygen mapping in the development and optimization of tissue-engineered scaffolds.