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The aim of the study was to evaluate in vitro the effect of native and deproteinized compact and spongy allogenic bone matrices on the characteristics of adipose mesenchymal stromal cells (ASC) in combined tissue engineering. Material and Methods. 24 samples of native and deproteinized compact and spongy bone were examined, which were exposed to mechanical treatment, modeling, followed by sterilization of the samples by ionizing radiation and bacteriological control of sterilization. Some of the samples underwent deproteinization. The characterized cultures of human ASC were used as test cultures to assess the interaction with the bone samples. The Cytation-5 fluorescent imager and Hoechst 3334 fluorochromes (BD Pharmingen™) and calcein (Calcein AM, BD Pharmingen™) were used to characterize the degree of adhesion, migration, and viability of ASC on bone matrix samples. Matrix cytotoxicity was evaluated by MTT assay on days 1 and 7 of extraction. Results. The bone matrix samples are characterized by the absence of cytotoxicity (rank 1). ASC demonstrated good adhesion and migration on any surface of the bone matrix and preservation of cell viability during 7 days of observation. Nuclei sizes of the cells adhered to the deproteinized bone matrix of the spongy structure increased by 25–30% compared to other samples. The cells on deproteinized bone matrix had greater size (the size of the cells from nuclei 8.8 to 11.5 μm, the average size of cells nuclei from an 86.3 μm to 129,0 μm, the average perimeter of the cells nuclei from 30.7 μm to 40.7 μm) than in the native bone matrix samples. Conclusion. The results of the study of various allogeneic bone matrices demonstrate that deep purification of the bone matrix determines the absence of cytotoxicity and the most favorable conditions for the adhesion, migration, proliferation and viability of ASC. Also makes it possible to use tissue engineering based on bone matrices of different structures. Deproteinized spongy bone matrices are best suited for this purpose.

Here we present the data comparing maturation of peripheral blood mononuclear cell-derived dendritic cells (DCs) isolated from healthy volunteers and Staphylococcus aureus-positive patients with chronic osteomyelitis. Dendritic cells were cultured in a standard maturation cell medium (RPMI-1640, supplemented with antibiotics, L-glutamine, 15% calf embryonic serum) added with interleukin-4 and granulocyte-macrophage colony-stimulating factor, followed by adding a stimulating factor cocktail containing interleukin-1β, tumor necrosis factor-α, interleukin-6, and prostaglandin E2. Dendritic cell maturation was analyzed by estimating visual characteristics under Zeiss ODSERVER.Z1 inverted microscope using Axiovision Rel.4.8 imaging software as well as light and phase-contrast microscopy at magnification of ×40, ×100, ×200, ×400, ×630. Dendritic cell immunophenotyping was carried out by using a panel of anti-human monoclonal antibodies: anti-CD80 FITC-conjugated, anti-CD86 (B7–2) PE-conjugated, anti-HLA-DR PC7-conjugated (all from Beckman Coulter, USA), anti-CD14 PerCP-Cy5.5-conjugated, anti-CD83 APC-conjugated, anti-CD40 PE-Cy7-conjugated (Becton Dickinson, USA) as well as isotype-matched control antibodies on the FACS Canto II f low cytometer (Becton Dickinson, USA). It was shown that while maturation dendritic cells derived both from patients or volunteers increased in size and underwent dendrite formation. Moreover, expression of CD86, CD83, CD80, and CD40 markers on dendritic cells derived from patients vs. volunteers was lowered. However, DC stimulation resulted in significantly increased percentage of DCs positive for CD83 DCs co-stimulation molecules CD86, CD80, CD40 chronic osteomyelitis. However, such differences found in immature DCs in both groups disappeared upon maturation, so that expression of the key markers on day 10 was maintained at close level. In particular, expression of CD83 differentiation marker and the CD80 co-stimulation molecule on DCs from patients vs. volunteers was increased stronger. Thus, a maturation potential in DCs isolated from patients with Staphylococcus aureus-caused chronic osteomyelitis was not impaired in vitro. The data obtained open up an opportunity to use dendritic cells as a natural adjuvant-substituting component for development of individualized vaccines in treatment and prevention of recurrent chronic osteomyelitis.

The purpose of the study is to assess the possibility of applying the integrated module as the basis of a celltissue equivalent for treatment of wounds of skin and soft tissues. In the frame of the set task the following problems were being solved: research of the spatial structure and architectonics of the surface of the developed base collagen-containing materials and their biocompatibility with cell cultures.Materials and methods. The study of a material which is a two-layer complex film, consisting of collagen and polysaccharide components was carried out. The collagen was separated from the dermis and was then impregnated with particulate demineralized bone matrix (DCM) according to the original methodology. For the purposes of the study the dehydrated material was created in the form of a film. Electron microscopic examination of surfaces was performed on scanning electron microscope JEOL JSM-IT300LV in high vacuum and at low values of probe current (< 0,1 nА). Studies to assess the viability of the cells cultivated on films of collagen material (tested for cytotoxicity and the adhesive capacity) were performed in vitro using strains of diploid human fibroblasts 4–6 passage. The culture condition was visually assessed using an inverted Leica microscope DM IL (Carl Zeiss, Austria), equipped with a computerizes program of control of culture growth (Leica IM 1000).Results. The data obtained in the study of the surface structure of the developed complex module showed that it seems to be promising as a basic component of the cellular-tissue system with its large number of structural formations for fixation of the cells and a well-organized barrier layer capable of vapor - permeability. Experiments in vitro confirmed the absence of toxicity of the material being studied in relation to the culture of dermal human fibroblasts, suggesting the possibility of creation on its basis of cell-tissue complex and further experimental studies in vivo.Conclusion. Thus it was experimentally confirmed that the physical characteristics of the developed integrated module satisfy the requirements for the materials for cultivation of cells. The absence of cytotoxicity on the model of a culture of dermal human fibroblasts allows to make a conclusion about a possibility of its use as the basis of cell-tissue equivalent. Preliminary results indicate the advisability of further experiments in vivo aimed at improving complex collagen-containing materials, the development of different ways of their application and clinical evaluation of the effectiveness in the treatment of wounds of various genesis.

Цель. Оценить выживаемость больных ХСН, страдающих СД 2; выявить параметры диабета, влияющие на течение и прогноз пациентов с недостаточностью кровообращения. Материалы и методы. Проведено исследование когорты дожития, которую составили 72 больных ХСН, страдающих СД 2. Судьба больных прослежена в течение 12 мес. У всех больных был собран анамнез и проведено физикальное обследование. Производилась оценка ФК ХСН. Выполнено Эхо-КГ. Исследовали уровень предсердного натрийуретического пептида (ПНУП). Результаты. Всего за 12 мес. наблюдения скончались 17 больных. Достоверно с выживаемостью связаны показатели тяжести ХСН, отражающие переносимость нагрузки (функциональный класс), клиническую характеристику (ШОКС) и нейрогуморальную активацию (ПНУП). Уровень ПНУП, более чем в 4 раза превышающий норму (т.е. более 8000 фмоль/мл), явился мощным предиктором неблагоприятного исхода в течение одного года. В группе выживших больные со ?стажем? СД более 10 лет составили 43% против 76% в группе скончавшихся. В группе больных с уровнем НвА1с менее 6,5% в половине случаев отмечались признаки хронической почечной недостаточности. При ХПН снижается активность почечной инсулиназы. достоверно на выживаемость оказывали влияние тяжесть ХСН и наличие уремической стадии диабетической нефропатии. На каждый балл увеличения ШОКС риск смерти в течение года возрастает на 25%. Наличие ХПН увеличивает риск смерти в 3 раза. Выводы. Почти 1/4 часть больных ХСН, страдающих СД 2, погибает в течение года. Существенное влияние на выживаемость оказывает исходная тяжесть ХСН. Наибольшее значение для прогноза среди характеристик СД 2 имеет диабетическая нефропатия. Ухудшение выживаемости отмечено уже на стадии микроальбуминурии. При наличии уремии риск смерти возрастает в 3 раза. Для быстрой оценки риска смерти в течение года можно ориентироваться на ШОКС и на наличие признаков ХПН. Для улучшения прогноза больных ХСН, страдающих СД 2, необходимо особое внимание уделять профилактике, ранней диагностике и лечению диабетического поражения почек.

Background: The quality of the wound healing at the donor site significantly determines the overall condition of the burn patient, the extent of wound fluid and protein losses, the severity of any systemic in-flammatory reaction, and the intensity of the pain syndrome. It is known that the stromal vas-cular fraction (SVF) has a beneficial effect on the healing of wound defects. This study is aimed at assessing the safety and effectiveness of the application of the SVF of autologous adipose tis-sue to stimulate wound healing of the donor site in patients with burns., Methods: This placebo-controlled clinical study included 38 patients with third-degree thermal skin burns. The patients underwent liposuction, enzymatic isolation of the SVF, and intradermal injection of the preparation into the wounds in the donor site, followed by tewametry, cutome-try, thermography and biopsy after 12 days. Quantitative indicators were compared using the Mann-Whitney test for unrelated groups and the Wilcoxon test for related groups. Spearman's rank correlation coefficient (RS) was used to assess the correlation., Results: Epithelization of the wounds in all patients was seen over an average area of 88 (84;92) %, there being no significant differences between the actual and the control wound sites for this parameter. Transdermal water loss in the test wound sites was 2 times lower than in the control sites ( P = 0.001). The wound donor sites regained their temperature distribution faster than the control sites ( P = 0.042). Histological preparations of the skin of the wound sites revealed that their epidermal layer was 19% thicker compared to the controls ( P = 0.043). It should be noted that five adverse events related to manipulations in the postoperative period were registered., Conclusion: Transplantation of SVF autologous adipose tissue into the wound area in most clinical cases proceeded without complications. The area of epithelialization of wound areas af-ter the introduction of SVF did not change, although a significant decrease in transdermal water loss was observed in the wound areas with an improvement in their thermoregulation and an increase in the thickness of the epidermis., Competing Interests: The authors declare that they have no competing interests., (© 2023 Iran University of Medical Sciences.)