Unlocking clotting mechanisms in caterpillar hemolymph for medical use

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Blood is a exceptional materials: it should stay fluid inside blood vessels, but clot as shortly as doable exterior them, to cease bleeding. The chemical cascade that makes this doable is nicely understood for vertebrate blood. But hemolymph, the equal of blood in bugs, has a really completely different composition, being notably missing in pink blood cells, hemoglobin, and platelets, and having amoeba-like cells referred to as hemocytes as an alternative of white blood cells for immune protection.

The caterpillar of another species of moth, the yellow-striped oakworm (Anisota peigleri). Image credit: Konstantin Kornev
The caterpillar of one other species of moth, the yellow-striped oakworm (Anisota peigleri). Image Credit: Konstantin Kornev

Just like blood, hemolymph clots shortly exterior the physique. How it does so has lengthy remained an enigma. Now, supplies scientists have proven in Frontiers in Soft Matter how this feat is managed by caterpillars of the Carolina sphinx moth. This discovery has potential purposes for human drugs, the authors mentioned.

“Here we show that these caterpillars, called tobacco hornworms, can seal the wounds in a minute. They do that in two steps: first, in a few seconds, their thin, water-like hemolymph becomes ‘viscoelastic’ or slimy, and the dripping hemolymph retracts back to the wound,” mentioned senior creator Dr Konstantin Kornev, a professor on the Department of Materials Science and Engineering of Clemson University.

“Next, hemocytes aggregate, starting from the wound surface and moving up to embrace the coating hemolymph film that eventually becomes a crust sealing the wound.”

Challenging to review

Fully grown tobacco hornworms, able to pupate, are between 7.5cm and 10cm lengthy. They solely include a minute quantity of hemolymph, which generally clots inside seconds, which makes it onerous to review with typical strategies.

For these causes, Kornev and colleagues needed to develop new strategies for the current examine, and work quick. Even so, the failure charge for the trickiest manipulations was monumental (as much as 95%), requiring many makes an attempt.

They restrained particular person hornworms in a plastic sleeve, and made a slight wound in one among every caterpillar’s pseudolegs by means of a window within the sleeve. They then touched the dripping hemolymph with a metallic ball, which was pulled away, making a hemolymph ‘bridge’ (about two millimeters lengthy and a whole lot of micrometers extensive) that subsequently narrowed and broke, producing satellite tv for pc droplets. Kornev et al. filmed these occasions with a excessive body charge digital camera and macro lens, to review them intimately.

Instantaneous change in properties

These observations prompt that in the course of the first roughly 5 seconds after beginning to movement, hemolymph behaved equally to water: in technical phrases, like a Newtonian, low viscosity liquid. But inside the subsequent 10 seconds, the hemolymph underwent a marked change: it now didn’t break instantaneously however shaped an extended bridge behind the falling drop. Typically, bleeding stopped utterly after 60 to 90 seconds, after a crust shaped over the wound.

Kornev et al. studied the hemolymph’s movement properties additional by inserting a 10-micrometer-long nickel nanorod in a droplet of contemporary hemolymph. When a rotating magnetic area brought about the nanorod to spin, its lag relative to the magnetism gave an estimate of the hemolymph’s capability to carry the rod again by means of viscosity.

They concluded that inside seconds after leaving the physique, caterpillar hemolymph adjustments from a low-viscous right into a viscoelastic fluid.

A superb instance of a viscoelastic fluid is saliva. When you smear a drop between your fingers, it behaves like water: supplies scientists will say it’s purely viscous. But because of very massive molecules referred to as mucins in it, saliva types a bridge whenever you transfer your fingers aside. Therefore, it’s correctly referred to as viscoelastic: viscous whenever you shear it and elastic whenever you stretch it.”

Dr Konstantin Kornev, Professor at Department of Materials Science and Engineering, Clemson University

The scientists additional used optical phase-contrast and polarized microscopy, X-ray imaging, and supplies science modeling to review the mobile processes by which hemocytes combination to type a crust over a wound. They did this not solely in Carolina sphinx moths and their caterpillars, but additionally in 18 different insect species.

Hemocytes are key

The outcomes confirmed that hemolymph of all species studied reacted equally to shear. But its response to stretching differed drastically between the hemocyte-rich hemolymph of caterpillars and cockroaches on the one hand, and the hemocyte-poor hemolymph of grownup butterflies and moths on the opposite: droplets stretched out to type bridges for the primary two, however instantly broke for the latter.

“Turning hemolymph into a viscoelastic fluid appears to help caterpillars and cockroaches to stop any bleeding, by retracting dripping droplets back to the wound in a few seconds,” mentioned Kornev. “We conclude that their hemolymph has an extraordinary ability to instantaneously change its material properties. Unlike silk-producing insects and spiders, which have a special organ for making fibers, these insects can make hemolymph filaments at any location upon wounding.”

The scientists concluded that hemocytes play a key function in all these processes. But why caterpillars and cockroaches want extra hemocytes than grownup butterflies and moths continues to be unknown.

“Our discoveries open the door for designing fast-working thickeners of human blood. We needn’t necessarily copy the exact biochemistry, but should focus on designing drugs that could turn blood into a viscoelastic material that stops bleeding. We hope that our findings will help to accomplish this task in the near future,” mentioned Kornev.

Source:

Journal reference:

Aprelev, P., et al. (2024) To seal a wound, caterpillars remodel blood from a viscous to a viscoelastic fluid in a couple of seconds. Frontiers in Soft Matter. doi.org/10.3389/frsfm.2024.1341129.

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