IntroductionTrophic ulcers are a common health problem, and there are numerous treatment methods. Irreversible damage in the skin, subcutaneous tissue, and fascia with long-term ulcer existence make standard autotransplantation inneffective. Skin grafts are often complicated by partial or complete rejection of skin flaps. The aim of this study was to examine the feasibility of using transplanted cultivated allogenic fibroblasts on the backing of a cellularless xenogenic fabric for wound healing.MethodsTransplantation of cultured embryonic fibroblasts on a backing of xenogenic tissue was used in the complex treatment of trophic ulcers for stimulation of regenerative processes. Decellularization xenogenic film was previously held. Then allogenic fibroblasts were cultivated on the surface of collagen-elastin matrix. Since 2013, we treated 12 patients with giant ulcers caused by the following: lymphedema (2 patients), vascular disease (3 patients), diabetes (2 patients), after injury (4 patients), and radiation ulcer (1 patient). Dimensions of ulcers were from 150 to 600 cm2. Duration of the lower limb ulcers ranged from 8 months to 10 years. For a number of years, all patients were on a complex therapy, which had not resulted in healing wounds. During the operation when excision of granulation tissue was performed, plastic wounds perforated with the ratio 1:2 autoskin. Xenogenic fabric with cultured fibroblasts was applied on top. In this case, xenogenic film protected the skin from drying, created optimal microclimate, and cultured fibroblasts stimulating regeneration and improving engraftment.ResultsThe first redress was held on the fifth day. In all cases, the results of engraftment skin grafts achieved maximum possible (100%) and optimal (90%). Complete epithelialization of the cell perforation was seen in five patients on the fifth day and three on seventh day after skin plastics. Average period of inpatient treatment was 20.7 days. All patients were discharged with healed wounds.ConclusionThus, the treatment of trophic ulcers can be successfully solved using advances in biotechnology. Transplantation of cultivated allogenic fibroblasts on a backing of cellularless xenogenic fabric shows good clinical results due to the stimulation of regenerative processes and creates the optimum environment for autotransplants.

IntroductionMicroelements, essential nutrients that are needed in small amounts including minerals such as calcium, zinc, iron and other vitamins (A, B, C, and etc.), are macronutrients necessary for a healthy life.The role of micronutrients in vivo is well known, and there are several publications that have examined the effects of micronutrients on genomic stability. Furthermore, a number of vitamins and microelements are substrates and/or cofactors in metabolic pathways, which regulate DNA synthesis and/or repair and gene expression.A deficiency in such nutrients may result in disruption of genomic integrity and alterations in DNA methylation patterns, linking cellular nutrition with change in gene expression. For example, lack of vitamin C is known to cause increased DNA oxidation and chromosomal damage. Vitamin A, as well as other micronutrients, have a protective effect, whereas higher concentrations are associated with increased DNA damage.Ubiquinone (coenzyme Q10) and dihydroquercetin are used in therapy as antioxidant compounds and electron carriers, which reduce lipid peroxidation of cell membranes. However, previous studies indicate that various ubiquinone analogs may cause a divergent effect on oxidative stress and oxidative phosphorylation.The aim of our study was to investigate the effect of vitamins A and C, coenzyme Q10, and dihydroquercetin on the proliferative potential of cultured human embryonic diploid fibroblasts (M-22).MethodsIn the first series of experiments, nontoxic concentrations of vitamins for the cells were identified using MTT assay.ResultsVitamins A and C, dihydroquercetin of 1μM, and coenzyme Q10 of 5μM were nontoxic for human skin fibroblasts. In the second series of experiments, cell cultivation was carried out with nontoxic concentrations. A vitamin C concentration of 1μM for 7 consecutive passages increased the proliferation index (PI) compared to the control. Thus, the average PI in the experiments was 2.3, whereas in the control, it was 1.7. Similar results were obtained when dihydroquercetin was added to the growth medium. However, further cultivation of cells in the presence of vitamin C decreased PI to 1.4, while the control value remained the same. Daily examination revealed no morphological changes in the cell culture, but the cell growth had slowed significantly. The use of vitamin A in a nontoxic concentration of 1 μM reduced PI to 0.7 in the first passage, so further culturing of human cells with vitamin A was stopped.ConclusionStudies examining the effect of different combinations of microelements on the proliferation of human diploid cells and the expression of specific proteins in them are still being conducted.

IntroductionBurns are an important public health challenge due to the frequency of getting burns in day-to-day life, occupational hazards, and catastrophes. Treatment of burns is complex and is associated with high morbidity and mortality. Duration and complexity of burn treatment require finding new ways of curing and rehabilitating burns. The result of burn treatment plays a significant role in post-traumatic status of a patient and his or her consequent adaptation in society. Chitosan is a natural safe non-toxic product compatible with human tissues, characterized by hydrosorbid, anticoagulant, antibacterial, and wound healing features. The study aims to show a clinical application of chitosan-pectin scaffold with cultured human skin fibroblasts in the treatment of deep burns.MethodsThe substrate was prepared by dissolving 3% chitosan in 0.5N acetic acid, which was then mixed with 3% solution of pectin dissolved in distillated water. Chitosan film was formed in a Petri dish for 20–24 hours at 20–25 °C. After drying the film, cultured allogeneic fibroblasts (patent number RK-25091) were seeded on its surface.ResultsThe results from an in vitro culture study showed that human allogeneic fibroblasts could adhere well and grow on the selected scaffold with a typical morphology. During autodermoplasty surgery, cultured allogeneic fibroblasts were applied on granulating wounds of 9 patients with IIIA to IVB degree burns and limited donor resources. Wounds treated with the fibroblastseeded scaffold among all patients provided the highest level of re-epithelialization (day 5), in comparison to cell-free scaffold (day 7) and untreated surface of wounds (day 10).ConclusionOur results indicate the potential use of chitosan for wound healing due to its allogenic fibroblast adherence to scaffolding as well as high epithelization. This warrants further studies on chitosan for use in wounds resulting from third and fourth degree burns.