Thermal injury healing in the context of neocollagenogenesis induction: preclinical randomized experimental study

Background. The healing of skin wounds having various etiologies is known to involve a multistep process characterized by certain intercellular interactions affecting dermal cells, their attachment, migration, and differentiation. Here, recovery is interpreted as the return of dermis to its original state. The fact is, however, that the dermal extracellular matrix (ECM) is structurally impaired, which suppresses the regulatory and repository functions of the dermis, leading to the formation of a scar that inhibits several biological functions in the affected area and causes aesthetic problems associated with mobility.Objectives. To evaluate the structural features of dermis during wound healing using a calcium-containing biodegradable implant.Methods. The study used 60 rats that were inflicted with a third-degree burn injury (partially damaged dermis). The selected animals were divided into two groups: experimental and control. On post-burn day 14, a calcium-containing biodegradable implant was administered to rats from the experimental group, while a sterile saline solution was used in the control group. Material was sampled at two months (74 days) and four months (134 days). In order to assess the morphological state of the burn area, its sections were stained with hematoxylin and eosin, according to Mallory and Van-Gieson. For the selective detection of collagens, immunohistochemical tests using antibodies to collagen types I and III (Abcam, England) were employed. To characterize dermal cells, the authors used antibodies to vimentin (LabVision, USA), as well as to СD-68 (cluster of differentiation 68), α-SMA (alpha-smooth muscle actin), CD-105 (cluster of differentiation 105), and VEGF (vascular endothelial growth factor) receptors (Abcam, England). The obtained results were processed using the Statistica 6 software (StatSoft, USA).Results. The administration of a calcium-containing biodegradable filler during the burn healing process was found to ensure local fibroblast activation with the formation of collagen types I and III. When the implant residence time was prolonged up to four months, an increase in the number of macrophages expressing CD-68 receptors was observed. Of note is that these cells retained their localization, while α-SMA-expressing cells were localized in both the superficial and deep dermal compartments. The number of cells expressing CD-105 and VEGF rose as well.Conclusion. The use of the biodegradable filler is found to be promising in terms of post-burn dermal regeneration, as well as providing a dermal ECM, whose collagen network composition and assembly are similar to the original. Here, macrophages act as the primary synthesis regulators of the dermal ECM and stimulate fibroblasts, which ensures re-epithelialization and angiogenesis of the inflicted area

Механизмы ангиогенеза при трансплантации тканеинженерных конструкций

There is a number of problems regarding bioengineered structures creation that require further study in the fi eld of regenerative medicine. One of the critical tasks that require solution is the fact that tissue engineered constructions, as a rule, are large, which signifi cantly limits the possibility of diffusion of nutrients and oxygen in them. Thus, the key task of fundamental medicine is to fi nd a technology for restoring the perfusion of the structures created. The article presents a modern overview of the mechanisms of angiogenesis and possible ways of its stimulation during transplantation of tissue engineered constructions.В области регенеративной медицины при создании биоинженерных конструкций имеется ряд проблем, нуждающихся в дальнейшем изучении. Одной из актуальных задач, требующих решения, является тот факт, что тканеинженерные конструкции, как правило, имеют большие размеры, что значительно ограничивает возможность диффузии в них питательных веществ и кислорода. Таким образом, ключевая задача фундаментальной медицины заключается в поиске технологии восстановления перфузии создаваемых конструкций. В статье представлен современный обзор механизмов ангиогенеза и возможных путей его стимуляции при трансплантации тканеинженерных конструкций

Морфологическая оценка качества децеллюляризации пищевода кры

The main aim of our investigation was analysis and optimization of the existing protocols of esophagus decellularization in rats, identification of parameters affecting the quality of decellularization and intactness of the native properties of esophageal extracellular matrix (ECM). Materials and methods. We developed a modified decellularization protocol based on detergent-enzymatic method using sodium deoxycholate and DNAse. Results. Morphological evaluation of the obtained decellularized matrices demonstrated the absence of cells and cell elements while mechanical properties of ECM were preserved. Conclusion. The developed protocol of esophagus decellularization is a potential basis for obtaining of tissue-engineered esophagus scaffold constructions.Основной целью нашего исследования явились анализ и оптимизация существующих протоколов децеллюляризации пищевода на модели крысы, определение параметров, влияющих на качество децеллюляризации и на сохранность исходных свойств ВКМ пищевода. Материалы и методы. Децеллюляризацию пищевода крысы выполняли с использованием оригинального и модифицированного протоколов, детергент-энзиматическим методом с применением дезоксихолата натрия и ДНКазы. Качество децеллюляризации оценивали с помощью рутинных гистологических методов исследования. Результаты. Морфологические методы оценки полученного децеллюляризированного матрикса подтвердили отсутствие ядер и клеток при сохранности трехмерности, архитектоники каркаса и биомеханических свойств ВКМ. Заключение. Разработанный протокол децеллюляризации пищевода крысы является перспективной основой для получения тканеинженерных конструкций пищевода

Dermal response to combined double filler administration

Aim. This experimental work presents a comparative assessment of the effect of hyaluronic acid and crystalline calcium hydroxyapatite on derma in separate and combined application setting.Materials and methods. The study used rats (30 animals) for subdermal injection of 0.05 ml medium. Group 1 was administered calcium hydroxyapatite (Radiesse (Merz, Germany)), group 2 received hyaluronic acid (Restylane (SabQ, Sweden)) and Group 3 — both preparations combined in ratio 1:1 (Mix). Results were evaluated 4 months after the filler injection. Sections were stained with haematoxylin–eosin, van Gieson’s and Masson’s trichrome techniques. Collagen types I and III were detected with polyclonal antibodies (Abcam, England). Fibroblasts were positively identified with vimentin (LabVision), macrophages — with CD68 (LabVision) tagging. Effect of extracellular matrix remodelling was studied with α-SMA actin (Abcam, England).Results. We demonstrate that separate filling of hyaluronic acid and calcium hydroxyapatite produces different response mechanisms, while their combined administration does not lead to a marked voluming of the dermal extracellular matrix.Conclusion. Combined administration of the two fillers under study may facilitate a prolonged effect of their combined action exceeding 4 months, due to the absence of fibrosis, complications and side effects

Status epidermis in introduktion of the drug «radiesse» and «luminaire»

Changes occurring which took place in epidermis skin after injection drugs «radiesse» and «luminaire» are analyzed in the experiment. Preparations introduced endodermal evaluation results conducted at 2, 4 and 16 weeks experiment to drug «radiesse» and at 2 and 16 weeks to drug «luminaire». Slices stained hematoxylin and eosin, by Weigert and Van-Gieson for selective detection DNA used reaction Feulgen. Distribution of total protein studied in preparations stained amido - black at pH 2.2, and cationic at pH 8.2. Spended the morphometry of cores were differentiated in layers epidermis and given assessment their optical density. It is shown that the introduction of fillers lead to increase thickness epidermis more than 1.5. The zone of germenative compartment increases mitotic activity in epithelial cells. Cells of granular layer increasing temp of protein synthesis having cationic charge. Revealed changes induced status DNA cell nuclei differentiated layers epidermis

MORPHOGISTOSCHEMICAL CHARACTERISTIC OF THE IMPLANT FROM POLYLACTIC ACID

Aim. The study was conducted for the characteristics of the morphoimmunohistochemical status of the implant from polylactic acid (PLA) located in the dermis.Materials and methods. The study was performed on rats (30 individuals) with subdermal administration of the drug in a volume of 0,05 ml. The results were evaluated after two weeks, 1 and 2 months after the injection. The sections were stained with hematoxylin and eosin, according to Van Gieson, with the help of Mallory and Masson reaction. Collagen fibers were studied after the staining with picrosirius red under polarized light. Monoclonal antibodies of CD 68 and antibodies to vimentin were used to determine the expression of immunohistochemical markers. Results. It is shown that the rate of collagen synthesis varies in different parts of the dermis. There is a sharp increase in the volume of the fibrous component around the implant, and there is only a tendency to increase in the synthesis in the implant itself. In this case, the increase in the volume of the extracellular matrix is associated with type I and III collagen.Conclusion. The detected activation of collagen synthesis by fibroblasts can be used as a regulator of the volume of the matrix, which is very important for the modern regenerative medicine

Morphometric characteristic of nuclear junction zone myometrium cells in adenomyosis

A morphometric study of cell nuclei intact junctional zone and in patients with adenomyosis. The material for the study is based on samples amputated during queens operations (47 cases), and autopsy material from apparently healthy women (6 cases). Wiring and sealing in paraffin. Measuring the optical density was nuclei (Feulgen reaction). The diameters of the cores were used to calculate the index of nuclear symmetry. We measure the amount of amorphous and fibrous components, as well as the total amount of hypochromic and hyperchromic nuclei. It was found that in patients with adenomyosis in the area junctional zone of endometrial stromal cell nuclei are characterized by an increase in the coefficient of ellipticity and absorbance. The nuclei of smooth muscle cells show pleomorphism and increase the index of the nuclear symmetry. The formed foci of adenomyosis identified unidirectional changes. It is possible that the nuclear symmetry index and the index of the optical density of the nuclei can be interpreted as molecular predictors of oncogenic transformation

СРАВНИТЕЛЬНАЯ ХАРАКТЕРИСТИКА ВЛИЯНИЯ ЭНДОГЕННЫХ И ЭКЗОГЕННЫХ НЕЙРОПЕПТИДОВ НА СЛИЗИСТУЮ ОБОЛОЧКУ ДВЕНАДЦАТИПЕРСТНОЙ КИШКИ COMPARATIVE CHARACTERISTICS OF MORPHOLOGICAL AND HISTOCHEMICAL OF THE INFLUENCE OF ENDOGENOUS AND EXOGENOUS NEUROPEPTIDES ON THE MUCO

; ГБУЗ «Детская краевая клиническая больница», Краснодар Работа посвящена изучению динамики морфологических и гистохимических преобразований, разы-грывающихся в слизистой оболочке двенадцатиперстной кишки (ДПК) в ответ на экзогенное введение ней-ропептидов и на нейропептиды, выделяющиеся эндогенно под влиянием транскраниальной электростиму-ляции (ТЭС). Материалом для исследования послужили кусочки ДПК, взятые от крыс, разделенных на три группы. Первая -контрольная, вторая -животные, подвергавшиеся ТЭС-терапии. Третья -это животные, которым вводили даларгин в дозе 24 мкг/кг массы тела каждые 12 часов в течение 11 дней. Изучены мор-фологические параметры слизистой ДПК и темп синтеза эпителиоцитами слизистой муцина. Оказалось, что трехдневная ТЭС-терапия приводит к увеличению толщины слизистой (в 2 раза), высоты ворсинок и глубины крипт. При этом глубина крипт нарастает от 144 ± 7,3 мкм (в контроле) до 284 ± 26,03 мкм. Бо-каловидные клетки ДПК увеличивают темп синтеза муцинов, и общая муцин продуцирующая активность резко увеличивается. При экзогенном введении даларгина, морфометрические параметры ДПК нарастают постепенно: первоначально увеличивается толщина слизистой (2-е сутки), затем высота ворсинок и глубина крипт (4-е сутки). Активация синтеза муцинов приурочена лишь к 4-м суткам наблюдения. На 11-е сутки введения даларгина и число бокаловидных клеток, и темп синтеза ими муцина приближаются к уровню контроля. Таким образом, эффект ТЭС-терапии -это резко выраженная гиперплазия слизистой и активация темпа синтеза муцинов бокаловидными клетками. Эффект экзогенного даларгина однонаправлен, но про-лонгирован во времени, а темп синтеза муцинов выражен менее активно. This paper studies the dynamics of morphological and histochemical changes that take place in the mucosa of duodenum in response to exogenous administration of neuropeptides and to neuropeptides endogenously evolved under the infl uence of transcranial electrostimulation (TES). Material for investigation are pieces of duodenum taken from the rats which were divided into three groups. First -a control, the second -the animals exposed TEStherapy. Third -animals which were administered with dalargin in a dose of 24 mg/kg body weight, every 12 hours within 11 days. Morphological parameters of duodenal mucosa and rate of mucin synthesis by mucosal epithelial cells are studied. It turned out that a three-days TES therapy increases mucosal thickness (2 ), the height of the villi and crypt depth . Thus crypt depth increases from 144 ± 7,3 mm (control) to 284 ± 26,03 microns. Goblet cells of duodenum increase a rate of mucin synthesis, and total mucin producing activity increases dramatically. When dalargin is administered exogenously, morphometric parameters of duodenum grow gradually: initially increases mucosal thickness (2nd day), then the height of villi and crypt depth (4 th day). Activation of mucin synthesis occurs only on 4 th day of observation. On 11th day after dalargin administration the number of goblet cells and the rate of mucin synthesis approach a control level. Thus, the effect of TES-therapy is a pronounced hyperplasia of the mucosa and activation of rate of mucin synthesis by goblet cells. The effect of exogenous dalargin administration is unidirectional, but prolonged in time, and the rate of mucin synthesis is less active