Optimal experimental design for efficient toxicity testing in microphysiological systems: A bone marrow application.

Area of Science:

• Biomedical Engineering
• Toxicology
• Cell Biology

Background:

• Microphysiological systems (MPS), or organ-on-a-chip technology, aim to mimic human organ environments for more predictive preclinical testing.
• The complexity of MPS necessitates careful management of technical factors to ensure reliable and scalable results.
• Assessing lineage-specific toxicity in the bone marrow (BM) is crucial for drug development.

Purpose of the Study:

• To develop and validate a systematic experimental design approach for bone marrow microphysiological systems (BM-MPS).
• To optimize the assessment of lineage-specific toxicity in BM-MPS, enhancing power, efficiency, and scalability.
• To demonstrate the utility of the developed approach for detecting drug-induced toxicities at earlier stages and lower doses.

Main Methods:

• Developed a systematic experimental design for a BM-MPS, focusing on assessing lineage-specific toxicity.
• Utilized multicolour flow cytometry for accurate cell type and maturity determination.
• Implemented a "repeated measures" design for increased scalability and robustness, incorporating an optimal arrangement of technical confounders.

Main Results:

• Validated the multicolour flow cytometry setup for precise cell characterization.
• Demonstrated that accounting for technical confounders in a mixed-model analysis increased statistical power.
• Successfully detected expected lineage-specific toxicities from olaparib and carboplatin treatments earlier and at lower doses.
• Performed sample size analysis to guide replicate number determination for various effect sizes.

Conclusions:

• The established design of experiments approach provides a foundation for reproducible in vitro analysis of BM toxicity using MPS.
• The generated lineage-specific toxicity data confirm the utility of this BM-MPS model for comprehensive toxicity assessment.
• This systematic approach is generalizable to other MPS applications, improving preclinical drug evaluation.