In 2014, during the first battles for Ukraine’s territorial integrity, it became clear that the military medical system was unprepared for new challenges. Above all, there was an acute shortage of effective hemostatic and wound-healing agents. At that time, the only high-quality hemostatic product available was Celox; however, its high cost significantly limited its widespread use. In addition, the product had a practical drawback – it was difficult to remove from the surface of a wound.

It was then that a group of researchers from the Institute of Cell Biology of NAS of Ukraine, led by Doctor of Biological Sciences Maxim Lutsik, developed a technology for purifying chitin from crustacean shells and producing chitosan with improved biocompatibility and effective hemostatic properties. In 2015, the research group received a Ukrainian patent for this development; however, it was not possible to launch production in Ukraine. The technology was rather complex and required the use of aggressive chemical reagents.

In 2022, following the start of the full-scale invasion of Ukraine by russian troops, the group led by Corresponding Member of the NAS of Ukraine Rostyslav Stoika, Head of the Department of Regulation of Cell Proliferation and Apoptosis at the Institute of Cell Biology of NAS of Ukraine, returned to this line of research. With grant support from the National Research Foundation of Ukraine (hereinafter – the NRFU, the Foundation), the researchers implemented the project “Anti-Inflammatory and Wound-Healing Agent Containing the Composition of the Cannabimimetic N-Stearoylethanolamine with Chitosan-based Hydrogel”.

The Foundation’s grant support made it possible to bring the medicine to the stage of technological development. The researchers formulated the medicine, conducted bioassays on laboratory animals, tested its biotolerance in vivo, and confirmed its safety.

Professor Stoika reflects: “Why this idea in particular? Because our defenders need an effective, affordable, Ukraine-made wound-healing agent. And we knew we could create one.”

A Wound-Healing Ointment

The essence of the project lies in the development of a multifunctional wound-healing medicine that combines everything necessary for the complex process of wound healing in a single ointment formulation.

The first component of the ointment is high-molecular-weight chitosan. It is responsible for stopping bleeding and, due to its viscosity, physically blocks damaged blood vessels. The second component is the lipid substance N- Stearoylethanolamine, or NSE. This substance has a wide range of beneficial properties: it reduces inflammation and pain, and supports cells in the damaged area. The third component is linseed oil which serves as a solvent for NSE. In addition, linseed oil is rich in Omega-3 and Omega-6 fatty acids, which function not only as antioxidants but also as immunostimulants. The fourth component is an antibiotic that suppresses wound infection. In field conditions, wounds are far from sterile, making this component essential. Moreover, the antibiotic performs another important role: owing to its negatively charged carboxyl group, it interacts with the positively charged amino groups of chitosan, thereby stabilizing the entire formulation.

“The know-how behind our medicine lies in incorporating NSE into its formulation and using linseed oil to dissolve it,” explains the PI. “This made it possible to combine a fat-soluble anti-inflammatory compound with the aqueous environment of a living organism.”

As a result, the researchers obtained a convenient ointment-like formulation that adheres well to the wound area, does not spread, offers a comprehensive therapeutic effect, and – most importantly – has no toxic side effects. A series of studies on laboratory mice confirmed these findings.

A Team from Across the Country

The development of the medicine was a collective effort that brought together research groups from several institutions across Ukraine.

Researchers from the Institute of Cell Biology of the National Academy of Sciences of Ukraine worked directly with the laboratory animals. They applied various versions of the treatment daily to the wounds of laboratory mice, photographed the injuries, monitored the healing process, recorded the research results, and also prepared ointment formulations and tested different recipe variants.

The anti-inflammatory lipid NSE and all associated research were handled by a group from the Palladin Institute of Biochemistry of NAS of Ukraine. Histological studies of wounds in laboratory animals were conducted by researchers from Danylo Halytsky Lviv National Medical University.

When There Is No Light or Heat in the Vivarium

The war disrupted the researchers’ work on a daily basis. The institute’s vivarium is located in a basement facility where maintaining the required temperature regime is challenging. During large-scale power outages, there was neither electricity nor sufficient heating. Researchers came in on weekends to feed and water the animals, check their condition, and clean the cages. They did so without reminders, time off, or additional compensation.

“I am sincerely grateful to all project participants, especially those who worked with the laboratory animals. It is demanding and highly responsible work, and my colleagues performed it with full dedication,” says Rostyslav Stoika.

Everything Begins with an Idea

Alongside the research results, Rostyslav Stoika willingly shares his own ‘algorithm for success’ in winning competitive research grants. He developed this approach over many years, including during nearly two years of work at biomedical centers in the United States.

The professor is convinced: “Fifty percent of success depends on an interesting and genuinely innovative idea. Moreover, such an idea is often not obvious – one has to work to arrive at it.”

Another 30 percent of success, in his opinion, comes from selecting the right experimental model which should be accessible, convenient, and at the same time appropriate for the objectives of the study. His experience in the US taught him that the global research community places the highest value on clinical studies involving real patients and allocates the greatest funding to them, followed by preclinical studies on laboratory animals, and only then studies based on cell cultures or computer models. Behind this hierarchy lies a simple principle: science must be useful for real life. The remaining 20 percent of success depends on the methods used and the people implementing the research.

The researcher consistently applies this approach in his own work.

Indicators of Success

For researchers, an important indicator of a successful study is publication in a high-impact scientific journal. While this article was being prepared, a paper was published in the internationally renowned journal “Wound Repair and Regeneration”, considered one of the leading journals in the field of wound healing. The article, entitled “Multifunctional Chitosan‐Based Ointment Supplemented With Cannabimimetic N‐Stearoylethanolamine for Wound Healing”, acknowledges the support of the NRFU without whose grant funding the development of this multifunctional wound-healing medicine would not have been possible.

From the Laboratory to a Soldier’s First-Aid Kit

The developed medicine has successfully passed laboratory testing and confirmed both its efficacy and safety. Today, the team is seeking partners willing to bring it into production.

 

Svitlana GALATA