Revitalization of decellularized or devitalized matrix scaffolds in tracheal tissue engineering typically involves seeding the autologous recipient cells or allogeneic cells under long-term cultivation. Objective: to study the capability of human nasal chondrocytes for colonization of devitalized scaffolds based on native human tracheal cartilage, with proinflammatory stimulation (cytokine) by adding Interleukin-1-beta (IL-1β) to the culture medium. Materials and methods. Scaffolds for tracheal tissue engineering were obtained from native human tracheal cartilage through devitalization and laser etching. The scaffold was revitalized by seeding the human nasal chondrocytes. Histological examination was performed after staining with hematoxylin and safranin-O, with further microscopy using a Nikon Eclipse L200 light microscope. X-ray microtomography was performed on a Phoenix nanotom m apparatus. Electron microscopy was performed on a Nova NanoSEM 230 setup. Results . There was statistically significant increase in the intensity of colonization (p = 0.0008) with nasal chondrocytes and stimulation of their migration activity (p < 0.0001) in the presence of IL-1β compared with the control groups. Conclusion. Addition of proinflammatory cytokine IL-1β (1 μg/ml) to the culture medium enhances volumetric seeding of devitalized cartilage scaffold with human nasal chondrocytes, allowing to create highly revitalized materials for tracheal tissue engineering.

Objective : to study the viability of a tissue-engineered graft (TEG) based on a devitalized tracheal scaffold (DTS) seeded with mesenchymal stromal and epithelial cells in an experiment on rabbits with assessment of cytocompatibility and biocompatibility in vivo . Materials and methods . Syngeneic mesenchymal stromal bone marrow cells (MSBMCs) and syngeneic lung epithelial cells of rabbit were obtained. The morphology and phenotype of the MSBMC culture were confirmed via immunofluorescence staining for CD90 and CD271 markers. Pulmonary epithelial cells obtained by enzymatic treatment of minced rabbit lung tissue were stained with CKPan, CK8/18 and CK14 markers characteristic of epithelial cells. The donor trachea was devitalized in three successive freezethawing cycles. Double-layer cell seeding of DTS was performed under static and dynamic culturing. Orthotopic implantation of TEGs was performed at the site of the anterolateral wall defect in the rabbit that was formed as a result of tracheal resection over four rings. Results were evaluated by computed tomography, histological and immunohistochemical analyzes. Results . A TEG implant, based on DTS, with bilayer colonization by cell cultures of rabbit MSBMC and epithelial cells was obtained. Three months after implantation, TEG engraftment was noted, no tracheal wall stenosis was observed. However, slight narrowing of the lumen in the implantation site was noted. Six months after implantation, viability of TEG was confirmed by histological method. Epithelialization and vascularization of the tracheal wall, absence of signs of purulent inflammation and aseptic necrosis were shown. The small narrowing of the lumen of trachea was found to have been caused by chronic inflammation due to irritation of the mucous membrane with suture material. Conclusion . A new model for assessing the viability of a tissue engineering implant when closing a critical airway defect was created. The developed TEG – based on DTS seeded (bilayer) by lung epithelial cells and BMSCs – was successfully used to replace non-extended tracheal defects in an in vivo experiment. The use of tracheal tissue-engineered graft for orthotopic implantation showed biocompatibility with minimal tissue response.

Проведение доклинических исследований для оценки безопасности и эффективности лекарственного средства является необходимым условием для его регистрации и внедрения в клиническую практику. Успех данных исследований во многом зависит от выбора животного и способа моделирования определенной патологии. В представленном обзоре методов анализируются данные по моделированию гинекологической патологии на лабораторных животных. В частности, рассмотрены методы воздействия, позволяющие моделировать синдром Ашермана, эндометриоз и хронический эндометрит. Поиск исследований производился в электронной библиотеке PubMed и базe клинических исследований Clinical Trials. В работе приводится характеристика использующихся хирургических вмешательств, физических, химических и инфекционных методов нарушения целостности матки и формирования в органе воспалительного и спаечного процессов. Рассматривается возможность моделирования эндометриоза при помощи стрессового воздействия. Оценена эффективность применения данных стратегий и возможность их дальнейшего использования. Анализ литературы показал, что наиболее оптимальным вариантом для моделирования повреждения матки считается использование крыс-самок в возрасте от 5 до 8 нед. Наиболее воспроизводимым способом имитации патоморфологических механизмов внутриматочной адгезии является повреждение эндометрия 95-процентным этанолом. Наиболее универсальным способом моделирования хронического эндометрита считается подшивание участков эндометрия к париетальной брюшине передней брюшной стенки. Preclinical studies are performed to assess safety and efficacy of a drug. They are a prerequisite for drug registration and implementation. Success of these studies depends on the choice of the animal and the method of modeling the pathology. This review focuses on modeling gynecological pathologies in preclinical studies. Specifically, the authors addressed experimental impacts for modeling Asherman’s syndrome, endometriosis, and chronic endometritis. The search for literature was conducted in PubMed and Clinical Trials databases. The review describes surgical interventions, physical, chemical, and infectious methods for affecting the uterus integrity and induction of inflammatory and adhesive processes in this organ. A possibility of modeling endometriosis by stress exposures is considered. Effectiveness of these strategies and prospects for their future use were evaluated. According to the analysis of reports the most suitable animal for modeling the uterine damage is 4-8-week-old female rats. The most reproducible way to activate mechanisms of pathomorphological intrauterine adhesion is chemical injury of the endometrium with 95% ethanol. The best approach in modeling chronic endometritis is endometrial and parietal peritoneum stitching.

Синтезированы наночастицы серебра с использованием для их восстановления и стабилизации арабиногалактана и диоктилсульфосукцината натрия. Средний гидродинамический размер наночастиц, определенный по данным фотонной корреляционной спектроскопии, составлял 30 нм, дзета-потенциал –34.04 ± 1.54 мВ. По данным метода электронной дифракции серебро в образце золя находится в металлической форме. Препарат наночастиц серебра проявлял антибактериальную активность в отношении условно-патогенных грамотрицательных (Escherichia coli) и грамположительных (Bacillus subtilis и B. coagulans) бактерий. Наночастицы серебра также обладали антифунгальной активностью в отношении штаммов фитопатогенных грибов рода Fusarium sporotrichioides и F. solani. Проведено исследование цитотоксической активности наночастиц серебра в отношении клеток гепатомы печени человека линии HepG2. Продемонстрировано ингибирующее действие наночастиц серебра в отношении метаболической активности и жизнеспособности опухолевых клеток. Средние относительные значения EC50 для наночастиц серебра составляли 1.5 ± 0.4 и 41.2 ± 3.9 мкг/мл. Препарат стабилизированных наночастиц серебра может найти применение в медицине в качестве потенциального антимикробного и противоопухолевого средства, а также в сельском хозяйстве в качестве средства подавления роста фитопатогенных грибов.

The aim of the study was to evaluate the effect on angiogenesis of a biocompatible, biodegradable material-derived scaffold implanted into rats and functionalized using a plasmid with a vascular growth factor gene. Methods. Experiments were performed on 24 female Wistar rats aged 2 months weighing 180-200 g. We investigated 1 cm x 1 cm flat scaffolds obtained by electrospinning from polycaprolactone functionalized scaffolds with a VEGF-165 plasmid (gene therapy drug, Neovasculgen) incorporated inside the fibers at two concentrations, low (0.005 mg/ml) and high (0.05 mg/ml). The sample and control were simultaneously implanted subcutaneously into two formed symmetrical pockets in the interblade zone. At 7, 16, 33, 46, and 64 days, the scaffolds were removed, and histological examination was performed; the tissue reaction was studied including morphometric evaluation of density and diameter of blood vessels in the implantation area, and the area of the image occupied by the material was measured. Results. Tissue rejection was absent after implantation of either control or modified material. When the material was exposed in vivo, besides resorption of the material, blood vessel number and diameter changed. As the Neovasculgen concentration in samples increased, a dose-dependent effect of angiogenesis stimulation became evident. Vascular density was increased by 46% (high concentration, 33 days) in functionalized matrices compared to the control. After cessation of the drug treatment, the vascular density approached the control values. Conclusion. The developed technique for functionalizing polymeric scaffolds by administration of a solution of the gene therapy drug, Neovasculgen, into microfibers provides a prolonged and dose-dependent effect on growth of blood vessels in the implantation zone.

Цель исследования - оценка влияния на ангиогенез конструкций из волокнистого поликапролактона, модифицированного плазмидой с геном сосудистого фактора роста, при имплантации крысам. Методика. Эксперименты выполнены на 24 крысах-самках Вистар в возрасте 2 мес, массой 180-200 г. В работе исследовали плоские каркасы размером 1 см х 1 см, полученные методом эмульсионного электроспиннинга из раствора поликапролактона. Материал каркасов витализировали плазмидой VEGF-165 (геннотерапевтический препарат Неоваскулген), введенной внутрь двух типов волокнистых материалов в разных концентрациях: низкой - 0,005 мг/мл, и высокой - 0,05 мг/мл. Образец и контроль (материал без витализации) одномоментно имплантировали подкожно в два сформированных симметричных кармана в межлопаточной зоне. Окружающие каркас ткани на 7-е, 16-е, 33-и, 46-е и 64-е сутки извлекали, проводили гистологическое исследование: изучали тканевую реакцию с морфометрической оценкой плотности распределения и диаметра сосудов в области имплантации, а также оценивали степень биодеградации волокнистого материала. Результаты. Признаков тканевой реакции отторжения при имплантации как контрольного, так и модифицированного материала не выявлено. Показано, что при экспозиции материала in vivo наряду с резорбцией материала происходят изменения количества и диаметра сосудов. Выявлен дозозависимый эффект стимуляции ангиогенеза при увеличении концентрации Неоваскулгена в образцах. Для витализированных материалов отмечено увеличение плотности распределения сосудов на 46% (высокая концентрация, 33-и сут) по сравнению с контролем. После прекращения воздействия препарата, плотность распределения сосудов приближалась к значениям в контроле. Заключение. Разработанная методика витализации полимерных каркасов с внесением раствора геннотерапевтического препарата Неоваскулген внутрь микроволокон обеспечивает пролонгированный и дозозависимый эффект на рост сосудов в зоне имплантации. The aim of the study was to evaluate the effect on angiogenesis of a biocompatible, biodegradable material-derived scaffold implanted into rats and functionalized using a plasmid with a vascular growth factor gene. Methods. Experiments were performed on 24 female Wistar rats aged 2 months weighing 180-200 g. We investigated 1 cm x 1 cm flat scaffolds obtained by electrospinning from polycaprolactone functionalized scaffolds with a VEGF-165 plasmid (gene therapy drug, Neovasculgen) incorporated inside the fibers at two concentrations, low (0.005 mg/ml) and high (0.05 mg/ml). The sample and control were simultaneously implanted subcutaneously into two formed symmetrical pockets in the interblade zone. At 7, 16, 33, 46, and 64 days, the scaffolds were removed, and histological examination was performed; the tissue reaction was studied including morphometric evaluation of density and diameter of blood vessels in the implantation area, and the area of the image occupied by the material was measured. Results. Tissue rejection was absent after implantation of either control or modified material. When the material was exposed in vivo , besides resorption of the material, blood vessel number and diameter changed. As the Neovasculgen concentration in samples increased, a dose-dependent effect of angiogenesis stimulation became evident. Vascular density was increased by 46% (high concentration, 33 days) in functionalized matrices compared to the control. After cessation of the drug treatment, the vascular density approached the control values. Conclusion. The developed technique for functionalizing polymeric scaffolds by administration of a solution of the gene therapy drug, Neovasculgen, into microfibers provides a prolonged and dose-dependent effect on growth of blood vessels in the implantation zone.

On an experimental model of chronic fibrotic liver damage (male rats Wistar (n-60), damage of CCl4, the duration of the experiment 90 days) it was studied the effectiveness of cell therapy for the correction of chronic liver failure. These rats were divided into 3 experimental groups: in the Ist-group (control, n=10) isotonic saline (650 mkl.) was injected; in the IInd-group (n=20) suspension of liver cells was applicated in a dose 8 - l0 x 10(6) cells; in the IIIrd-group (n=30) suspension of liver cells and bone marrow cells (mesenchymal stromal cells) in ratio 5:1 were used as cell associates on microparticles intjectable heterogeneous biopolymer hydrogel "SpheroGEL" (cell-engineering design) in common dose 8 - l0 x 10(6) It was ascertained that in the 2nd and in the 3rd groups the accelerated normalization of disturbed liver functional indices (ALT, AST, ALP) took place - to 30 days, but in the control group only to 90 days. The reliable differences in rats ofnormalization offunctional indices were absent between the IInd and the IIIrd groups. But in 90 days by using special histological dyeing it was found out that defibrotic processes in liver tissue were more expressed in the IIIrd group in comparison with the IIIrd group. Received results were consequence of prolonged vital activity of cells (liver cells and mesenchymal stromal bone marrow cells) into cell-engineering designs, which were transplanted in the IIIrd group. The obtained effect can be explained by that the developed cell-engineering designs provide adequate conditions for prolonged vital activity of the transplanted cells.

On an experimental model of chronic fibrotic liver damage (male rats Wistar (n=60), damage of CCl 4 , the duration of the experiment 90 days) it was studied the effectiveness of cell therapy for the correction of chronic liver failure. These rats were divided into 3 experimental groups: in the I st group (control, n=10) isotonic saline (650 mkl.) was injected; in the II nd group (n=20) suspension of liver cells was applicated in a dose 8-10×10 6 cells; in the III rd group (n=30) suspension of liver cells and bone marrow cells (mesenchymal stromal cells) in ratio 5:1 were used as cell associates on microparticles injectable heterogeneous biopolymer hydrogel «SpheroGEL» (cell-engineering design) in common dose 8-10×10 6 cells . It was ascertained that in the 2 nd and in the 3 rd groups the accelerated normalization of disturbed liver functional indices (ALT, AST, ALP) took place – to 30 days, but in the control group only to 90 days. The reliable differences in rats of normalization of functional indices were absent between the II nd and the III rd groups. But in 90 days by using special histological dyeing it was found out that defibrotic processes in liver tissue were more expressed in the III rd group in comparison with the III nd group. Received results were consequence of prolonged vital activity of cells (liver cells and mesenchymal stromal bone marrow cells) into cell-engineering designs, which were transplanted in the III rd group. The obtained effect can be explained by that the developed cell-engineering designs provide adequate conditions for prolonged vital activity of the transplanted cells.

For the last decade many experimental and clinical data about the study and application of regenerative medicine methods in the maxillofacial surgery were accumulated. For better bone regeneration mesenchymal stem cells are often used. Considering the general wariness of researchers in some aspects of cell therapy, methods of study of mesenchymal stem cells and the technologies of its clinical application are constantly being upgraded. This review will consider methods of tissue engineering used to regenerate bone tissue defects in maxillofacial surgery.

Before using MSC transplantation in the clinic to conduct preclinical studies MSCs to animals with acute and chronic pancreatitis. Work out the timing and dose of MSCs. The rationale of MSCs transplantation for the regeneration of damaged pancreatic tissue. The essence of the experiments is to establish the existence of common pathogenetic mechanisms for the development of pathological processes and sanogenesis toxic damage of pancreatic tissue. The study was work out in the rat model of acute and chronic pancreatitis, to explore beneficial and adverse effects of allogeneic stem cells for regenerative-reduction processes. For cell transplantation using allogenic stromal cell fraction of bone marrow, the cell suspension was injected at a dose of 2 x 10(6) and 5 x 10(6) cells.