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publicado em 11/09/2013 às 11h27:00
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Suture stem cells accelerates wound healing

Unicamp was developed material capable of accelerating the regeneration of tissues in the intestinal fistulas

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Foto: Antonio Scarpinetti/Unicamp
Imagem: Divulgação/Unicamp
Professor Angela Cristina Malheiros Luzo, guiding research Infographic of the research process that uses stem cells in sutures to accelerate wound healing
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Professor Angela Cristina Malheiros Luzo, guiding research
Infographic of the research process that uses stem cells in sutures to accelerate wound healing

The results surprised even the researchers who followed the evolution of an intestinal fistula - an open wound caused by a failure of healing still of unknown origin and difficult to treat. On the third day after application of a novel suture enriched with stem cells, noted that the area of ​​the wound size had decreased and nearly closed (75%). This is well above the regeneration obtained with existing conventional resources and implementing a long and complicated recovery in patients with this type of problem in humans, which can last from eight to ten weeks.

In the study, mesenchymal stem cells were responsible for the miracle that accelerates the regeneration of tissues in test animals (rats). They are in human body because they possess the ability to differentiate (turn) in several other cell types and may initiate the formation of new tissue, which explains the obtained result. In medicine, were first described in the 70s and since then, researchers have sought ways to profit from them, which is not so simple, as shown by other studies in the world and experiments produced at Unicamp, to discover the best way to transform them into other tissues and implant them in the body.

The average recovery of rat intestinal fistulae in the conventional treatment, it is necessary to close the treatment of humans (up to ten weeks).

Mesenchymal cells can be turned into other cells (those lacking the wound site) or released substances that have led to decreased inflammation and better vascularization in the area and activated the stem cell's own tissue in the affected area, so that multiply there, says Angela Luzo, about the effect on the healing process observed during the accelerated masters research conducted by biologist Bruno Volpe Bosch. The effects caused by them will be studied by the Multidisciplinary Group of Cellular Therapy Unicamp, the thesis of the author of the work, should also happen when clinical studies, or human trials.

The great innovation of the research was not the use of stem cells, but the methodology developed to inject it into the fistula through a suture enriched with mesenchymal cells, patented by Inova Unicamp Innovation Agency. According to the researchers, the simple application of the cells at the wound site, as demonstrated in experiments that did not show the same effect healing.

The search joined successfully: a suture joint used in medicine for decades, a human surgical glue (fibrin), biological sealant produced from the blood plasma, which has no contraindications or cause reaction in the patient; and millions of human mesenchymal stem cells, able to accelerate healing. With the application of the wire, we observed a 90% recovery [mean] the area affected by fistula, after 21 days, and in some animals the wound closed completely, evaluates Bruno.

In other studies conducted worldwide, according to the Brazilian study, we tested the application of stem cells directly on the wound (Spain), but without the result of wound healing, and producing a thread like (United States), produced by another process, but in limited quantities - at Unicamp, the researchers managed to cell adhesion on a wire over a foot, applied after two days of preparation, but there is an expectation that it survives for seven days.

The Brazilian group had assessed the effectiveness of the adhesion ability of stem cells in fibrin glue in an undergraduate research project conducted by the student Larissa Berbert, under the guidance of Professor Paul Kharmandayan, Department of Surgery, FCM, and teacher Angela Luzo, hematologist. This line of research emerged from a class in the discipline of plastic surgery, performed in 2005 by plastic surgeon Stocchero Ithamar, who questioned the ability of stem cells to adhere to suture.

Years later, the project worked because of Unicamp, in the laboratory, it was possible to grow stem cells, apply them to the suture, and especially keep them alive, in sufficient quantity so that they enter into action in the area injury. Images captured with the aid of a microscope show cells applied to the filaments which intertwine the suture completely taken by green color, marking each one, all alive, ready to begin the process of healing observed.

<b> Display </ b>

On average, 2% of patients undergoing surgery in the intestine develop this type of fistula that was the basis for the studies, but this number could reach 20% for high-risk patients, those who are taking immunosuppressive drugs, debilitated by cancer or are carriers of certain specific diseases. In the conventional treatment, in most cases, an attempt is made to direct suture of the fistula, which is quite complicated to make, or there is a nutritional support for patients and expected to close the fistula, which may take, sometimes 40 days, said Dr. Joaquim Bustorff-Silva, professor and coordinator of the Department of Surgery of the FCM, which participates in the research group.

When accessible medicine, the new technology will enable a simple treatment and should reduce the recovery time of patients. This greatly diminished the need for hospitalization, complications because of fistulas, but basically, will greatly reduce the time for solving the problem, evaluates the teacher. The open wound degrades the health of the patient, causes weight loss, dehydration, among other problems.

In addition, two results highlight the study: there would be no need to assess the compatibility of the stem cells used in suture for patients, as occurs today in certain therapies, and there were no signs of rejection or inflammation in the body relation to the implanted cells. In experiments performed by Unicamp, sutures were enriched with human stem cells for application in guinea pig intestinal fistulae (rats), in a procedure known as xenotransplantation.

For now, the treatment is still expensive solution, but with potential application in various medical procedures, particularly in patients at risk, as a way to speed healing and reduce complications in surgical procedures, according to the researchers. The wire can be used either enriched preventive treatment of such problems of these and other types of fistulas.

<b> Recovery occurs in less time </ b>

The healing of fistulas in rats sutures enriched with mesenchymal stem cells occurred, on average, 15 days after the procedure, more rapid evolution quite different from what was observed in the other two groups of guinea pigs in the experiment - those who had natural recovery from injury and those who underwent procedures direct application of stem cells at the wound site.

We know that this cell type [mesenchymal] releases several factors stimulating growth, substances that enhance healing, which decrease the inflammatory process in the region and thus can be applied to other types of surgeries and treatments, says hematologist Angela Cristina Malheiros Luzo, professor of postgraduate Sciences Surgery, Faculty of Medical Sciences, guiding the research.

In the experiment, all rats had intestinal fistulae of the same size. The wounds were monitored daily with the help of a computer program capable of evaluating per square millimeter, regeneration of the tissues of the affected area.

According to the author, the biologist Bosch Bruno Volpe, the group of guinea pigs that received no treatment, after 21 days, showed little progress after almost double the expected time for healing. In the other group, who received the stem cells, similar to that carried out by Spanish researchers, the recovery was 70% (the area affected by fistula). With the application of the wire, in the same period, the average recovery was 90%, explains. Furthermore, three of the nine mice that received the suture enriched in fact had complete healing during the observed period.

The graph shows the evolution of fistula closure in three groups of guinea pigs, impresses, as the photos show complete recovery of the wound tissue in those who received the wire with mesenchymal cells. On the third day, the result is almost equal to that obtained after 21 days, with the application of stem cells at the site of the fistula, says the researcher.

The direct application of the cells without the wire, an experiment was conducted in Spain and played Unicamp for comparison. There, researchers performed the work facing a possible treatment for Crohn's disease, a chronic inflammation of the intestine, of autoimmune origin and unknown. Symptoms and treatment depend on each case, but is the common occurrence of abdominal pain, diarrhea, weight loss and fever. Currently, there is no cure, but procedures to alleviate the complications faced by patients.

The results obtained by the research demonstrates the importance of the suture enriched developed at Unicamp research to the process of recovery and prevention of intestinal fistulae. More than that, the same methodology can be used in other types of similar procedures, which increases the potential for use in medicine, according to researchers at the Multidisciplinary Group of Cellular Therapy.

This research group at Unicamp, and researchers of the Faculty of Medical Sciences (FCM), also involves the Faculties of Mechanical Engineering (FEM), Chemical Engineering (HFE), plus the Institutes of Biology (IB), Chemistry (IQ) and Physics (IFGW), and the National Institute of Science and Technology in Biomanufacturing (Biofabris) and the National Synchrotron Light Laboratory (LNLS).

<b> Manufacturing </ b>

In the study conducted at Unicamp, the mesenchymal stem cells were extracted from fat. This type of cell exists in all tissues, but to extract, manipulate, fat is the easiest source, the researcher explained Angela Luzo. A bone marrow or cord blood would be other sources of this material.

From about 20 ml to 30 ml of fat, in which there are about one million MSCs, the method allows to multiply them in the laboratory and at the end of 20 days, there will 4000000-5000000 them, ready for the process manufacturing wire enriched. The human fibrin glue, used in production, has been enhanced to improve the adherence of these fibers suture (see the art). Doctors use it to contain bleeding, especially when the points do not account alone to stanch the blood.

In practice, to absorb the wire enriched with cells, the body receives the drug which accelerates proliferation of tissue at the wound site and that accelerates healing.

Caught the suture, put in culture, drip a million cells in it and apply the glue so that they are adhered. Once impregnated material, not loose and they are alive, explains Bruno, remembering that there were three main problems to be overcome: the cells die in contact with the glue, which does not resist both the surgical procedure (fabric application) and that there were enough of them to activate the process of regeneration of the area affected by the injury.

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Suture with stem cells    suture    stem cells    wound cicratização    mesenchymal stem cells    fat cells    intestinal fistulae    Bosch Bruno Volpe    health   
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