This exploratory one-year proposal investigates a new frontier for the fundamental understanding and application of clathrate hydrates in the stabilization of biological systems. Stabilization of biological systems, or biopreservation, is of major importance for the storage and preservation of cells and proteins in biology, biotechnology, medicine, and food technology. Clathrate hydrates are inclusion compounds formed from the hydrogen bonding of water molecules that enclose small, mostly non-polar compounds. The benefits of employing clathrate hydrates over current stabilization approaches, namely lyophilization (removal of water) and cryopreservation (cold storage), are the preservation of biological systems at fully hydrated conditions and the ability for stabilization at temperatures above the normal freezing point of water.

The intellectual merit of this novel study is to identify suitable clathrate hydrate formers to stabilize biological materials and to perform viability studies on biological systems to demonstrate the suitability of clathrate hydrate in their stabilization. The experimental measurements will be performed in a custom-designed cell suitable for hydrate formation and biological studies. Measurement of hydrate formation conditions will concentrate on hydrate formers that are benign to biological systems (cells and proteins) and have equilibrium formation above the normal freezing point of water (0?aC). Some of the hydrate formers to be considered in this study include mixtures of xenon and carbon dioxide with tetrahydrofuran and ethylene oxide. Measurements will be performed in both neat water and biological media. These measurements will serve as the basis to study the stabilization of biological systems with clathrate hydrates, which will be initially verified by the activity and viability of microorganisms and animal cells. This exploratory study is transformative, as it introduces novel concepts in the area of clathrate hydrates and new approaches for the stabilization of biological systems.

The broader impact component of the project is the investigation of a unexplored area of research with potential applications in the stabilization of cellular and protein systems, in the development of new food processing and preservation technologies, in the encapsulation of small organic agents, and in the mechanism for anesthesia and hibernation. The project will demonstrate the unique properties of clathrate hydrates in biology, which is an area beyond its well-established areas of research and applications (i.e., energy, storage, and environment). This project will also include training of a graduate student, who will work closely with the PI and his collaborator, Prof. Y. Martin Lo at the University of Maryland-College Park.

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Colorado School of Mines
United States
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