Revolutionary Research Challenges Standard Model of Electroporation and Opens Door to New Possibilities

Electric fields can create holes called “pores” in biological membranes. This process is called electroporation, and it’s an important technique for treating medical conditions and food items.

Two scientists, Dr. Carlos Marques and Professor Jan Behrends, have done some research recently which suggests that something we’ve known for decades may not be true anymore. Dr. Marques works in Lyon, France, while Professor Behrends works at the University of Freiburg in Germany – they worked together on this project.

Marques came up with a challenge that will help us learn more and use computers to test new theories. His findings were published in the Proceedings of the Academy of Sciences of the United States of America (PNAS). The results could make it easier for medicines and treatments to get into our cells.

Sparks of Change

When a powerful electric current is used, the molecules that make up the outer layer of cells (called lipids) get broken apart. This causes “electropores” which stay open only for a very quick time. While the electropores are open, water and other things like drugs, RNA, or DNA, can pass through into the cell.

The layer of lipids, which is really thin measuring only 0.000005 millimeters, doesn’t need a very high voltage to create a high field strength. At a voltage of just 0.1 volts across the membrane, it becomes 20 million volts per meter; more than enough for air to spark. However, this needs to be direct current voltage. Alternating current like those created by cell phones don’t cause pores. Even though scientists already know how this works, they’re still trying to make it better so that it can help with gene therapy by introducing genetic material into cells. To do this right, they need to understand what happens when electric fields come in contact with cells for sure.

Proving Electricity-Induced Pores with Careful Measurements

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In the 1970s, scientists created a theory that said electric fields can cause tiny pores to open up in lipids. But they had difficulty proving this theory through experiments because it was hard to see these tiny pores and they needed to do a lot of tests in order to have reliable result. This is tricky because the openings caused by electricity are really different compared to those caused by proteins.

Scientists have come up with a way to measure tiny electric currents that help detect the formation of pores. Pores are small openings on a cell’s membrane which charge particles, like salt, can flow in and out from. To do this, they create artificial lipid bilayers between two electrodes using Teflon plates with holes around 0.1 millimeters wide. The problem is that only one membrane is formed at a time and it breaks easily during tests with strong electric fields.

Discovering Pore Formation Through the Microelectrode Cavity Array

The research team used a special chip that had lots of tiny holes. This made it possible to create layers made from lipids more easily and quickly, compared to past methods. It’s called the microelectrode cavity array (MECA). The MECA was created by Jan Behrends’ group and was sold by Ionera Technologies GmbH – a business founded in 2014.

Eulalie Lafarge from the Charles Sadron Institute at University of Strasbourg and Dr. Ekaterina Zaitseva from the Freiburg research group invented a device that allowed them to create tons of tiny membranes quickly. And they used it to measure how strong the electric current had to be to form those microscopic pores.

The experiment showed that the electric field does not make it easier for pores to form by increasing the strength of the field, like was previously thought. Instead, doubling the power of the field only helps a little bit – not 4 times as much. This implies that there is a totally new way in which electric fields can form pores. It’s all due to how polar molecules (water) shift their orientation when they’re around an electric field.

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The results of an experiment were repeated with different levels of oxidation in the cell membrane. This is important because oxidation helps to keep cells working properly as we age and it can also be linked to diseases like Parkinson’s and Alzheimer’s. Scientists want to know more about this process and plan on doing further tests, including looking at the process through a microscope.

A group of scientists recently published an article titled: “Activation energy for pore opening in lipid membranes under an electric field”. This article explains the process of how pores can be opened in a lipid membrane when it is exposed to electricity. The paper was published on 7 March 2023, and can be found with the DOI: 10.1073/pnas.2213112120.