Using cellular technologies, the Ukrainian doctors have learned to restore the damaged limbs. The official medicine still does not recognize this method. But the facts are self-evident.

The traditional “Chestnut Race” was held on the last Sunday of May, which is a Kyiv Day. This year, the ilaya clinic medical staff will run together with about 20 soldiers from ATO Zone, whom doctors recommended to amputate mutilated limbs. But thanks to the efforts of ilaya employees they will be running on their own healthy feet and waving with healthy arms.

In modern war about 50% of wounds are represented by the affected limbs from mine actuations, artillery shells, and bullet. 5-7% of such injuries are accompanied by the bone defects. Traditional medicine applies in case of bone defects with a size from 5 cm the amputation or prosthesis. Ilaya clinic produces “pre-bone” of cells in laboratory conditions which is being transplanted into a bone defect site. Moreover, these are the fragments of 10-12 centimeters in size, and sometimes up to 15 cm.

The “Biotech-Rehabilitation of The Injured” Project was launched on the basis of the clinic in September, 2014. Since then, it engaged about 55 persons, 30 of whom underwent complete treatment.

Despite the obvious ilaya achievements, the Ukrainian traditional medicine is not willing to recognize this method, which based on the relatively new “stem cells” concept. 15-20 years ago, the world science was actively trying to introduce the cellular technology into medical practice as a panacea for all diseases. But it turned out that the use of stem cells can bring both benefit and harm. As a result, Europe and United States had all the activities on cellular technologies production and their medical application severely regulated and bureaucratized. Ukrainian experts were able to make a breakthrough in this field of limbs combat injuries treatment due to three components: war, lack of regulation, as well as cell technology study over the past 15 years.

Mine injuries

Donetsk can be rightly considered the centre of cellular technologies in Ukraine. In 1999, the director of the Institute for Emergency and Reconstructive Surgery (IERS) Vladimir Gusak suggested that stem cells can be used in the treatment of mine injuries, which are accompanied by severe burns: skin for grafting can be grown in the laboratory. “We were the graduates of Biology Faculty of Donetsk National University, when received an invitation to participate in the project. We were immediately forwarded to N. K. Kuznetsov’s Institute of Developmental Biology in Moscow, where we just “ripped off” the cultivation technology of keratinocyte layers,” Roman Vasilyev and Dmitry Zubov tell the biologists.

However, we failed to replicate the technology in Donetsk at once. “Empirically, we were changing the conditions, course of processes and, finally, succeeded”, the experts say, adding that the establishment of the biological laboratory at a Burn Centre lasted until 2003.

Since the mine injuries often involve complex fractures and bone deformities, the experts were involved to treat such patients from Donetsk Research Institute of Traumatology and Orthopedics (DRITO) at M. Gorky’s Donetsk National Medical University (DonNMU). In 2004, the heads of the two institutions decided to combine forces. “It all started from scratch. I and Dmitry Zubov were searching in literature, studying embryology to understand what to do to make the cell bone-like”, says Vladimir Oksimets, Orthopedic Traumatologist of the highest category, MD.

Stem cells, which trigger the development of all subsequent cells of the human body tissues, have been described in the early 60-ies of XX century and became known as bone lining cells. But how to determine which of the cells, into what tissue, will be produced: bone, cartilage, blood vessels, muscles, or skin? The European laboratories use chemicals for converting (induction) of bone lining cells into bone tissue cells. But in this case the cells stop dividing on the 19th day and no fracture can become solid during such a short period of time.

The discovery of Donetsk scientists was the use for the bone lining cells induction of endosteal and periosteal cells to be cultivated together. They are placed on a special implant, while transplantation the cells continue to divide until they fill the missing bone tissue volume, union with own bone, and then stop dividing.

At the end of 2004 the bone transplantation using cell technology was held in Donetsk for the first time. The first patient was a man, whose fracture failed to union within three years. The next patient had the same problems observed for five years. Then, the patients included miners with severe occupational injuries: one had almost severed arm at a shoulder level – was restored; the other had a traumatic heel defect – was “grown”; moreover, this was done for the first time in the world.

Within a period from 2004 to 2014 about 50 operations were carried out in Donetsk, related to stem cells: 32 post-injury transplantations of limbs and about 20 transplantations of the femoral head.

Promising cell

In 2010 two lawyers – Aleksey Shershnev and Aleksey Kotkovsky – decided to launch a medical business. “I studied the possibilities in this area and found the promising trend: cell technology,” Shershnev explains his choice. Partners decided to create a traditional multifunctional clinic, a separate area of which was to be regeneration – this segment had almost no competition.

ilaya Clinic was established in May 2012. The pride of the medical establishment has become a laboratory equipped with the latest technology. The cost of construction of such facilities, which regulates everything from air circulation to the staff movements, is Euro 4000-5000 per square meter. The sum of $ 1.5 million received as a loan from Ukreximbank was spent on the purchase of equipment, including incubators with the oxygen control from Binder. “The oxygen content in the atmosphere and human body differs significantly. The oxygen is maintained in incubators at a level same as in human tissues. Therefore, the implant transplanted is better adapted to the human body, and cells retain the genome stability”, biologists explain.

Stem cells have been described back in the early 60-ies of XX century

ilaya laboratory meets GMP standards used in assessing the pharmaceutical industry, and is the only laboratory in Ukraine of this level. But, as it turned out, this was not an argument for the Ukrainian officials. “We’ve tried for five times to obtain a license to work with cellular technologies. Only on the sixth time, in July 2013, the Coordination Centre for Transplantation of Tissues, Organs and Cells at the Ministry of Health granted it to us. This was the sixth one license in Ukraine”, Shershnev says. By that time, the total cost of lab amounted to $ 3.5 million.

War became a turning point

“When the war broke out in the east of Ukraine, I arrived to Kiev and explained to Shershnev the injuries which will be observed among the wounded persons. It will be a repetition of the war in Afghanistan: broken bones, no bones, non-union fractures. Just those cases where it is possible to use our cell technology”, says Vladimir Oksimets, a graduate of the Kirov’s Leningrad Military Medical Academy. We decided to develop the bone regeneration. Another reason for this was the lack of risk to the patient: traditional medicine already prescribed him an amputation, and cellular technology gives a chance to save the limb.

The problem was that the cellular regeneration is an expensive process. The support was provided by Shershnev’s friend and countryman from Nikolaev – IT-company owner David Arakhamia. “We used to play together in a rock band. His volunteer PeoplesProjects was implementing a variety of projects for the soldiers. David supported the “Biotech – Rehabilitation of the Injured” Project and launched fundraising on the project portal”, Shershnev says.

The first operations were 80% funded by Arakhamia’s Company. But following the first results, people believed in it. Today, the project has already collected nearly UAH 12.4 million: UAH 200-500 from individuals, hundreds of thousands of hryvnia donated from the companies, there are transactions from Canada, US and Europe. But fundraising is continuing: clinic is accepting new wounded persons and the average cost of the operation is US $ 15 000-20 000.

Over the year and a half there were as many transplantations carried out as for the previous 10 years. Along the way, the clinic specialists introduced several innovations. First of all, to obtain bone material it is now sufficient for biologists to have 5 cm3 of bone marrow from the patient’s iliac bone, which does not cause substantial harm to the patient. Second, the implantation technique improved: in the lack of a large part of the bone the defect site has a transplant placed, grown in the laboratory. Then, the surgery is performed to fix it.

Third, in addition to the two types of cells that form bone tissue, biologists have learned to receive and apply on the bone matrix a third type of cell – endothelial cells that form the vascular system of the bone. “In Donetsk we have been dealing with injuries of 2-4 cm, and there the graft blood supply had a time to form from the surrounding tissues and own living bone. In battle traumas, when it is necessary to close the bone defect with a size of 10-15 cm, the vascular cells in the graft are essential, they help forming a new vascular system in the transplant, “integrate” into the vascular system of own bone and union with the existing bone fragments”, Oksimets explains.

In addition, an unprecedented operation was carried out to restore the bones of the skull in 23-year-old guy, who after an accident had almost half of the temporal bone removed. Zubov, Vasilyev and neurosurgeon Vladimir Gritsik decided to take this incredible step. “The bones of the human skull and neck have a different origin than the limb bones. They cannot be mutually replaced; to confirm the theory an experiment was conducted in mice. Then, the patient skull bone was taken from the hospital storage. It was cleaned out completely of formalin and turned into the bone matrix cultivated with the patient’s cells. After a few months the living bone implant grew fully together with the other bones. Today, no traces of the operation can be seen under the hair”, biologists say.

Health and Business

The first results of operations were presented by Oksimets at the Regenerative Medicine Conference in Leipzig in 2013. The audience did not believe. Already in the lobbies the surgeon was explained that in Europe the grants to biologists for such studies range from Euro 6.5 million to 12 million for three years and those receiving grants have no need to hurry. In Ukraine, there are no such funds, and our specialists conduct researches within a couple of years: time elapsing from the start of experiment to the clinical trial.

Europe has only four officially registered cell products authorized for use: two products for the restoration of articular cartilage, one – to restore the corneal epithelium and therapeutic agent – to correct a complex genetic disease. No progress in the work on the recognition of new technology by the European regulator observed. USA takes time either with the registration of new technologies in the United States. The leading journal Regenerative Medicine published only two articles on treatment of blast injuries: in 2013 – the authors were from ilaya clinic staff and in 2014 – experts authorship from the Walter Reed National Military Medical Centre (Maryland, USA). It is not entirely correct to compare these works, because in Ukraine the regeneration is performed using patient’s own cells, and in the United States – from donor, i.e. other person’s cells.

ilaya experts intend to present the treatment results in soldiers in summer and fall of 2016: first in Sviatogorsk where Donetsk Medical University is going to host a conference for orthopaedist-traumatologists, then in Munich at the meeting of the European Society of Trauma and Emergency Surgery, and then in Rome at a meeting of the International Society of Orthopaedic Surgery and Traumatology.

Unfortunately, in Donetsk the institutions experts patented the utility model only – a declarative patent, which does not provide copyright protection for an invention. The first patent is always issued in the country of origin, and since the system is integrated throughout the world, it will be recognized in all states. Clinic attorneys have collected all the documents relating to the regeneration of bone, and have already submitted to the State Department of Intellectual Property of Ukraine. Application was accepted with no comment.

“It is a long-term procedure, involving the degree of novelty analysis, reconciliation of the technical data from all the patent reference books worldwide. In case of overlapping it is necessary to refine, adjust the nuances. Only then the claim is being derived and patent issued”, Shershnev explains. “It is not allowed to patent the treatment method; otherwise no one would be able to hold any single operation. But it is possible to patent the technology of creation, methods for performing certain operations, tissue – engineered products, and so on…”

Ilaya’s current license to work with stem cells allows applying all the methods of treatment associated with cellular technologies. But the clinic is committed to working within the framework of civilized rules, which requires not only guidelines and patents, but clinical trials as well that will allow registering a new treatment method and make it official, available for the entire country. Ilaya repeatedly filed documents for clinical trials of bone regeneration techniques to the Coordination Centre of the Ministry of Health, but without effect.

Patents and clinical trials – is recognition of the particular specialist or group’s contribution to the science, an opportunity to train other doctors, create own school in medicine, as well as the opportunity to receive royalties for using own intellectual property in other clinics. It is the ability of physicians and biologists to work in native country and receive a decent pay. And it is the possibility of the state medical tourism development.

Today, Ukraine allocates money for the prosthetics in soldiers who lost limbs. One bionic prosthesis costs UAH 2-3 million. In 2015 UAH 150 million were allocated, of which only UAH 18 million were utilized. “Had no time to execute the documents, or executed incorrectly”, Forbes’s source from the Ministry of Social Policy complains, who requested anonymity.

Ilaya can preserve many damaged limbs requiring amputation and prosthetics. “Biotech-Rehabilitation of the Injured” Project has saved the state more than UAH 200 million on social payments and about the same amount of funds required for the purchase of prostheses. The state budget for 2016 provides for the possibility of financing such operations. But it is May already, and the program has not started yet, so the volunteers continue raising funds on their own.