Potential of using cultivated dermal fibroblasts on the biodegradable polymeric matrices for treating skin damages in the experiment

ФИБРОБЛАСТЫЭКСПЕРИМЕНТЫ НА ЖИВОТНЫХБИОМЕДИЦИНСКИЕ ИССЛЕДОВАНИЯБИОДЕГРАДАЦИЯСОЕДИНИТЕЛЬНОЙ ТКАНИ КЛЕТКИКРЫСЫ ЛИНИИ WISTARКОЖА /ПОВРЕЖДДЕКСТРАНА ФОСФАТПОЛИЛАКТИДРАНЫ ЗАЖИВЛЕНИЕКЛЕТОК ТРАНСПЛАНТАЦИЯКЛЕТОЧНАЯ ТЕРАПИЯЦель. Оценить возможность применения культивированных дермальных фибробластов на полимер- ных биодеградируемых матрицах для лечения повреждений кожного покрова в эксперименте. Материал и методы. Исследование выполнено на 90 крысах линии Wistar массой тела 180-200 г, с чистыми ранами диаметром 2,5 см. Было сформировано 6 групп – по животных в каждой. Культуру дер- мальных фибробластов выделяли из кожи новорожденных крыс; для накопления необходимой биомассы клеток проводили субпассирование. В 1 группе (контроль) клеточные биопрепараты не применялись. В группах 2-6 для лечения экспериментальных ран были использованы биодеградируемые матрицы (фосфат декстрана либо полилактид) как без клеточного биопрепарата, так и с содержанием культивированных фибробластов в количестве от 250?103 до 700?103 клеток на один раневой дефект. Выводы о влиянии дермаль- ных фибробластов на полимерных биодеградируемых матрицах на раневой процесс делали на основании сравнительного анализа результатов компьютерной планиметрии и морфологического исследования. Результаты. На всех этапах (7, 14, 21, 28 сутки) эксперимента минимальная скорость заживления была зарегистрированы в группе 1. Шестая группа, в которой в рану вносили полилактид, содержащий 700?103 клеток, характеризовалась максимальными – относительно остальных групп – значениями скоро- сти заживления. В микропрепаратах 6 группы на 21 сутки эксперимента отмечена полная эпителизация ра- невого дефекта. Выявлено наличие сильной прямой корреляционной связи между количеством трансплан- тированных клеток и скоростью заживления раневых дефектов на 7 сутки (?=0,923), 14 сутки (?=0,924), а также 21 сутки эксперимента (?=0,914). Заключение. В условиях эксперимента установлено, что локальное применение культивированных дермальных фибробластов на полимерных биодеградируемых матрицах (фосфат декстрана и полилактид) ведет к достоверному сокращению сроков эпителизации раневых дефектов.Objectives. To evaluate the possibility of using of cultivated dermal fibroblasts on the biodegradable polymeric matrices for treating skin damages in the experiment. Methods. The study was performed on 90 rats of Wistar line (180-200 g weight) with pure wounds (D-2,5cm). Six groups were formed, 15 animals per each group. The culture of dermal fibroblasts was isolated from the skin of newborn rats; to accumulate the required biomass of cells subpassage was carried out. The cellular biological products were not applied in group 1 (control). Biodegradable matrices (dextran phosphate or polylactide) were used for treating wounds in the groups 2-6 as matrices without cellular biological product and matrices with cultured fibroblasts (250?103 to 700?103 cells per wound). Conclusion about the impact of biodegradable polymeric matrices with dermal fibroblasts on the wound healing was made on the basis of a comparative analysis of the results of digital planimetry and morphological studies. Results. At all stages (7, 14, 21, 28 days) minimal rate of healing was registered in the first group. The sixth group, where polylactide containing 700?103 cells was added in the wound, is characterized by the biggest values of the healing rate – with respect to other groups. Complete epithelization of the wound defects in the micropreparations of the sixth group was marked by the 21st day of the experiment. The strong direct correlation between the number of transplanted cells and wound rate healing was detected on the 7th day (?=0,923), the 14th day (?=0,924), and the 21st day of the experiment (?=0,914). Conclusion. In the experiment it has been found out, that local application of the cultivated dermal fibroblasts on biodegradable polymer matrices (dextran phosphate and polylactide) results in a significant reduction of terms of the wound epithelization

РЕГУЛЯТОРНЫЕ СВОЙСТВА КВЕРЦЕТИНА В КУЛЬТУРАХ МЕЗЕНХИМАЛЬНЫХ СТВОЛОВЫХ КЛЕТОК

The resistance to the oxidative stress, the proliferative activity, the immunophenotype stability, and the ability to differentiate in the adipogenic direction of MSCs from adipose tissue of rats after the incubation of cells in the presence of quercetin has been investigated. The cultivation of MSCs in the presence of 100 nM – 100 mM quercetin enhances the antioxidant defense system activity and the resistance of cells to the oxidative stress, decreases the intracellular ROS content (hydrogen peroxide), and inhibits the adipogenic differentiation of cells. At concentrations of 10–100 nM, polyphenol has a stimulating effect on the proliferative activity of MSCs and does not affect the immunophenotype of cells. At concentrations of 10–100 mM, quercetin decreases the proliferative activity of the cells and changes the expression of markers of multipotent MSCs CD29, CD44, and CD90.Приведено изучение устойчивости к окислительному стрессу, пролиферативной активности, стабильности иммунофенотипа и способности к дифференцировке в адипогенном направлении МСК ЖТ крыс в разные сроки после инкубации клеток в присутствии кверцетина в широком диапазоне его концентраций. Установлено, что культивирование МСК ЖТ в присутствии 100 нМ – 100 мкМ кверцетина увеличивает активность системы антиоксидантной защиты и устойчивость клеток к окислительному стрессу, снижает внутриклеточное содержание АФК (перекись водорода) и ингибирует дифференцировку клеток в адипогенном направлении. В концентрациях 10–100 нМ полифенол оказывает стимулирующее влияние на пролиферативную активность МСК ЖТ и не влияет на иммунофенотип клеток, при концентрациях 10–100 мкМ наблюдается снижение пролиферативной активности и изменение экспрессии маркеров мультипотентных МСК CD29, CD44 и CD90

Фурье-преобразование в сферических системах как инструмент решения физических задач структурной биологии

Applications of the most common adaptation of Fourier analysis in spherical coordinate systems used to solve a number of problems in structural biology, namely, flat wave decomposition (flat waves are represented as spherical functions decomposition), are herein considered. Arguments in favor of this decomposition are compared with other decompositions in superposition of special functions. A more general justification for the correctness of this decomposition is obtained than that existing today. A method for representing groups of atoms in the form of a Fourier object is proposed. It is also considered what opportunities give such a representation. The prospects for the application of Fourier analysis in structural biophysics are discussed.Рассмотрено применение наиболее распространенной адаптации Фурье-анализа в сферических системах координат для решения ряда задач структурной биологии, а именно: разложения по плоским волнам (при этом плоские волны представляются в виде разложения по сферическим функциям). Приводятся аргументы в пользу этого разложения в сравнении с другими разложениями по суперпозициям специальных функций. Получено более общее обоснование корректности данного разложения, чем существующие в настоящее время. Предложен способ представления групп атомов в виде Фурье-объекта и рассмотрены его возможности. Обсуждаются перспективы применения Фурье-анализа в структурной биофизике

Влияние аллогеной трансплантации мезенхимальных стволовых клеток жировой ткани на ноцицептивную чувствительность и репаративные процессы при экспериментальной травме ахиллова сухожилия у крыс

On the model of Achilles tendon injury in rats, the antinociceptive and regenerative effect of allogeneic transplantation of various doses of adipose-derived mesenchymal stem cells (ADMSCs) into the lesion area was studied. It was found that netiher of tested regimens of allogeneic transplantation of ADMSCs contributed to a decrease in the edema of the injured limb. Among the studied regimens, only a twofold administration of 0.25 • 106 ADMSCs exhibited an antinociceptive effect. Administration of 0.50 • 106 ADMSCs promoted the accelerated onset of neovascularization of tendon tissue, while enhancing inflammation and the formation of granulation tissue. Double transplantation of 0.25 • 106 ADMSCs led to accelerated reorganization of collagen fibers, later neovascularization, however, there was an absence of inflammatory infiltrate, lipomatosis, and massive formation of granulation tissue in the lesion area.На модели травмы ахиллова сухожилия у крыс изучен антиноцицептивный и регенеративный эффект аллогенной трансплантации различных доз мезенхимальных стволовых клеток жировой ткани (МСК ЖТ) в область повреждения. Установлено, что аллогенная трансплантация МСК ЖТ ни в одном из тестируемых режимов не способствовала уменьшению величины отека травмированной конечности. Среди исследуемых режимов только двукратное введение 0,25 • 106 МСК ЖТ оказывало антиноцицептивное действие. Введение 0,50 • 106 МСК ЖТ способствовало ускоренному началу неоваскуляризации ткани сухожилия, одновременно усугубляя воспаление и образование грануляционной ткани. Двукратная трансплантация 0,25 • 106 МСК ЖТ приводила к ускорению реорганизации коллагеновых волокон, более поздней неоваскуляризации, однако наблюдалось отсутствие воспалительного инфильтрата, липоматоза и массивного образования грануляционной ткани в области травмы

Induced pluripotent stem cells. Obtaining, properties and application prospects in biology and medicine

This review dedicated to induced pluripotent stem cells discovery of which became one of the outstanding achievements of biomedical science at the beginning of 21th century. These cells are in fact a artificially induced analogue of embryonic stem cells. They are the progenitors of all cells contained in organism of animals and human. The pool of embryonic stem cells forms in fertilized cell and they are involved in extremely complex processes of organogenesis. The position of these cells in general and of induced pluripotent stem cells in particular in hierarchy of cells in organism is described.The main properties of these cells, similarities and differences between embryonic and induced pluripotent stem cells, prospects for their practical use in biology and medicine including treatment of human diseases are considered.The mechanisms of induced dedifferenciation in both unipotent cells, for example fibroblasts and keretotinocytes, and multipotent cells as hemopoetic or mesenchimal stromal stem cells are described.In this review the information on used in the world approaches for obtaining of induced pluripotent stem cells is presented, the ways of reprogramming different in the methods of delivery of genetic material into the cells and conditions of their cultivation during dedifferentiation process and efficacy. As an example, description of all dedifferentiation cycle of human fibroblasts in induced pluripotent stem cells is provided. Besides, the data on unique properties of induced pluripotent stem cells what allowed to study ontogenesis processes, pathophysiology of various diseases both in vitro and in animals' models to be obtained using induced pluripotent stem cells and also on approaches of these cells application for medicine screening when they are synthesized and tested are given.</jats:p

Fourier transformation in spherical systems as a tool of structural biology

Applications of the most common adaptation of Fourier analysis in spherical coordinate systems used to solve a number of problems in structural biology, namely, flat wave decomposition (flat waves are represented as spherical functions decomposition), are herein considered. Arguments in favor of this decomposition are compared with other decompositions in superposition of special functions. A more general justification for the correctness of this decomposition is obtained than that existing today. A method for representing groups of atoms in the form of a Fourier object is proposed. It is also considered what opportunities give such a representation. The prospects for the application of Fourier analysis in structural biophysics are discussed.</jats:p

Clinical and roentgenological evaluation of the status of periodontal tissues of laboratory animals in the application of mesenchymal stem cells

The article examines the clinical and roentgenological changes in the periodontal tissues of laboratory animals when mesenchymal stem cells (MSC) are used.The aim of the study is to create a model of experimental periodontitis and identify the characteristics of clinical and radiological changes in periodontal tissues when applying a biomedical cell product based on allogeneic mesenchymal adipose stem cells (AT MSCs).During the examination of the clinical and radiological changes in the periodontal tissues ofexperimental animals with formed bone defects filled with AT MSCs, it was found that the mucous membrane regeneration time in the surgical area was comparable in all main groups of animals. Postoperative gum recession was observed in the control group animals. The significant differences between the clinical pictures in groups I–IV during all observation periods after surgery were not revealed. However, the restoration process signs in the post-resection area found during the roentgenological examination in the groups using osteoinduced MSCs, as well as a mixture of MSC cultures and osteo-induced MSCs, were most pronounced, which is confirmed by the bone mineral density.The experimental periodontitis model, which could be used for assessing the bone tissue restoration processes of a labioratory animal, was developed. Thus, the use of collagen membranes with a suspension of allogeneic osteo-induced AT MSCs cultures, as well as membranes with a suspension of a mixture of allogeneic and allogeneic osteo-induced AT MSCs in the ratio of 1:1 allows achieving higher bone tissue recovery rates.</jats:p

Morphological changes in periodontal tissues in laboratory animals using mesenchymal stem cells

The article presents the analysis of the morphological changes of the periodontal tissues of laboratory animals using mesenchymal stem cells (MSCs). The goal of the study is to create a model of experimental periodontitis and to identify the characteristics of morphological changes in the periodontal tissues using a biomedical cell product based on the allogeneic adipose tissue of MSCs (AT MSCs). The application of a mixture of AT MSCs and osteoinduced AT MSCs (in the ratio of 1:1) allows reducing the time of bone defect regeneration in comparison to that of bone tissue regeneration when AT MSCs and osteoinduced AT MSCs are used separately, which is expressed in the filling of the bone defect with a fibroreticular osteogenic tissue, as well as with a muscle tissue one month after surgery. In 2 months, in the defect area filled with a collagen membrane with a mixture of AT MSCs and osteoinduced AT MSCs, the initial signs of the formation of trabecula of bones were detected, which is evident of a more comprehensive osteosynthesis process compared to the blood clot use.</jats:p