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Hydrogel tissue construct-based high-content compound screening.

Vy Lam¹, Tetsuro Wakatsuki

¹Department of Physiology and Biotechnology and Bioengineering Center, Medical College of Wisconsin, Milwaukee, WI, USA.

Journal of Biomolecular Screening

|December 3, 2010

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Researchers developed a 3D hydrogel tissue construct (HTC) assay to better screen drug compounds. This novel system accurately measures mechanical properties and physiological parameters, identifying H-1152 as a promising candidate with minimal side effects.

Area of Science:

Biomedical Engineering
Pharmacology
Cell Biology

Background:

Current cell-based assays lack in vivo relevance due to 2D or suspension culture limitations.
Accurate assessment of drug effects requires evaluating mechanical and physiological properties in a more physiologically relevant context.

Purpose of the Study:

To develop and validate a high-throughput 3D hydrogel tissue construct (HTC)-based assay system.
To quantify mechanical properties and multiple physiological parameters of cells within a 3D construct.
To screen compounds for their ability to modulate HTC contractility and assess associated physiological effects.

Main Methods:

Developed an in vitro, high-throughput 3D hydrogel tissue construct (HTC) assay.
Quantified HTC mechanics using an automated device.

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Assessed physiological status via spectroscopy-based indicators read on microplate readers.

Screened four compounds (rotenone, cytochalasin D, 2,4-dinitrophenol, H-1152) for effects on HTC contractility, actin integrity, mitochondrial membrane potential (MMP), and viability.

Main Results:

All four tested compounds dose-dependently reduced HTC contractility.
Rotenone exhibited toxicity, 2,4-dinitrophenol dissipated MMP, and cytochalasin D reduced F-actin and viability.
The Rho kinase inhibitor H-1152 effectively reduced HTC contractility with minimal observed side effects.

Conclusions:

The developed HTC-based assay system provides a more physiologically relevant platform for drug screening.
This system enables the assessment of compound-induced changes in tissue contractility and underlying physiological mechanisms.
H-1152 emerged as a promising candidate for modulating HTC contractility with a favorable safety profile.