In a sharp and pointy world, wound healing is a critical and marvelous process.
Despite a tremendous amount of scientific study, many outstanding mysteries still surround the way in which cells in living tissue respond to and repair physical damage.
One prominent mystery is exactly how wound-healing is triggered: A better understanding of this process is essential for developing new and improved methods for treating wounds of all types.
Using an ultrafast, ultraprecise ultraviolet laser, a team of physicists and biologists at Vanderbilt University has taken an important step toward understanding the nature of these trigger signals. Their new insights are described in a paper titled "Multiple mechanisms drive calcium signal dynamics around laser induced epithelial wounds" published Oct. 3 by the Biophysical Journal.
Previous research had determined that calcium ions play a key role in wound response. That is not surprising, because calcium signaling has an impact on nearly every aspect of cellular life. So, the researchers -- headed by Professor of Physics and Biological Sciences Shane Hutson and Associate Professor of Cell and Developmental Biology Andrea Page-McCaw -- targeted cells on the back of fruit fly pupae that expressed a protein that fluoresces in the presence of calcium ions. This allowed them to track changes in calcium ion concentrations in the cells around wounds in living tissue (as opposed to the cell cultures used in many previous wound response studies) and to do so with an unprecedented, millisecond precision.
The team created microscopic wounds in the pupae's epithelial layer using a laser that can be focused down to a point small enough to punch microscopic holes in individual cells (less than a millionth of a meter). The laser's precision allowed them to create repeatable and controllable wounds. They found that even the briefest of pulses in the nanosecond to femtosecond range produced a microscopic explosion called a cavitation bubble powerful enough to damage nearby cells.
"As a result, the damage the laser pulses produce is quite similar to a puncture wound surrounded by a crush wound -- blunt force trauma in forensic terms -- so our observations should apply to most common wounds," said first author Erica Shannon, a doctoral student in developmental biology.
The researchers were testing two prevailing hypotheses for the wound-response trigger. One is that damaged and dying cells release proteins into the extracellular fluid which surrounding cells sense, causing them to boost their internal calcium levels. This increased calcium concentration, in turn, triggers their transformation from a static to a mobile form, allowing them to begin sealing off the wound. The second hypothesis proposes that the trigger signal spreads from cell to cell through gap junctions, specialized intercellular connections that directly link two cells at points where they touch. These are microscopic gates that allow neighboring cells to exchange ions, molecules and electrical impulses quickly and directly.
"What is extremely exciting is that we found evidence that cells use both mechanisms," said Shannon. "It turns out cells have a number of different ways to signal injury. This may allow them to differentiate between different kinds of wounds."
The experiments revealed that the creation of a wound generates a complex series of calcium signals in the surrounding tissue:
"Once we understand these trigger mechanisms, it should be possible to find ways to stimulate the wound healing process in people with conditions, like diabetes, that slow down the process or even to speed up normal wound healing," said Hutson.
Graduate student Aaron Stevens, research assistant Wes Edrington, undergraduate researcher Yunhua Zhao and graduate student Aroshan Jayansinge also contributed to the research, which was funded by National Institute of Health grants R21AR068933 and 5T32CA119925 and National Science Foundation grant REU 1560035.
Materials provided by Vanderbilt University. Original written by David Salisbury. Note: Content may be edited for style and length.
Disclaimer: DoveMed is not responsible for the accuracy of the adapted version of news releases posted to DoveMed by contributing universities and institutions.
References:
Erica K. Shannon, Aaron Stevens, Westin Edrington, Yunhua Zhao, Aroshan K. Jayasinghe, Andrea Page-Mccaw, M. Shane Hutson. (2017). Multiple Mechanisms Drive Calcium Signal Dynamics around Laser-Induced Epithelial Wounds. Biophysical Journal. DOI: 10.1016/j.bpj.2017.07.022
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