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Liposomes pp 377-391 | Cite as

Intracellular ATP Delivery Using Highly Fusogenic Liposomes

  • Sufan ChienEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 605)

Abstract

Healthy cells must maintain a high content of adenosine triphosphate (ATP) because almost all energy-requiring processes in cells are driven, either directly or indirectly, by hydrolysis of ATP. During ischemia or hypoxia, reduced blood flow or disturbed oxygen supply results in the disrupted balance of energy production and utilization, and depletion of high-energy phosphates is the fundamental cause of cell damage. Direct intravenous infusion of high-energy phosphates, such as adenosine triphosphate (ATP), has not produced a consistent result because strongly charged molecules like ATP normally cannot pass the cell membrane in sufficient quantities to satisfy tissue metabolic requirements. Furthermore, the half-life of free ATP in blood circulation is very short, limiting its efficacy as a bioenergetic substrate.

We have developed a new technique for intracellular delivery of high-energy phosphate into normal or ischemic cells by using specially formulated, highly fusogenic, unilamellar lipid vesicles that contain magnesium-ATP. In vitro studies indicated a rapid fusion with the endothelial cells, protection of endothelial cells, and cardiomyocytes during ischemia. In vivo studies have shown enhanced full-thickness skin wound healing in various animal models. This technique has the potential to reduce or eliminate many detrimental effects caused by ischemia or hypoxia.

Key words

Ischemia Hypoxia Intracellular delivery ATP Liposome Energy Shock Endothelial cells Phospholipids Wounds 

Notes

Acknowledgments

This study was supported in part by NIH grants HL64186, DK74566, and AR52984. We thank Drs. Benjamin Chiang and William Ehringer for their experimental contributions in cell culture and preliminary formulation of ATP-vesicles, Dr. Qunwei Zhang for his cytokine studies, and Dr. Jianpu Wang for his wound management in some rabbits. The author also wishes to thank Mr. Drew Chochran of Avanti Polar Lipids for his contribution in the section of Optimizing ATP-vesicles formulation, Mr. Robert Reed for his scanning EM examination, and Ms. Ming Li for her HPLC determinations

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of SurgeryUniversity of LouisvilleLouisvilleUSA

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