Трансплантация тканевых эквивалентов в лечении некоторых повреждений кожи

Chronic ulcers are a common and socially significant problem worldwide. Autodermoplasty is the gold standard treatment for chronic ulcers. However, it is not always possible to perform this surgical procedure for a rather large group of patients, due to some reasons, which include high risk of autodermotransplant rejection, lack of donor material, and patient’s unwillingness to undergo surgery with an often unpredictable result. A potential solution to the problem is to use skin equivalents from allogeneic donor material. The use of allogeneic (donor) human cells makes it possible to fill the deficit of the patient’s donor resources and close wound without causing additional injury to the patient. This paper provides an overview of the application of foreign and domestic biomedical cell products in clinical trials and real clinical practice. We draw conclusions on the efficiency of the considered biomedical cell products in the treatment of chronic ulcers, evaluate the conducted research, and make recommendations on the most efficient use of allogeneic dermatotropic biomedical cell products.Хронические раны являются распространенной и социально значимой проблемой во всем мире. Золотым стандартом лечения хронических ран является аутодермопластика, однако ее выполнение не всегда возможно у довольно большой группы больных, в том числе из-за высокого риска отторжения аутодермотрансплантата, дефицита донорского материала, нежелания пациента подвергать себя хирургической операции с часто непредсказуемым результатом. Возможным решением проблемы является использование эквивалентов кожи из аллогенного донорского материала. Использование аллогенных (донорских) человеческих клеток позволит восполнить дефицит донорских ресурсов пациента, выполнить закрытие раневого дефекта без нанесения пациенту дополнительного повреждения. В настоящей статье представлен обзор применения в клинических исследованиях и реальной клинической практике зарубежных и отечественных биомедицинских клеточных продуктов. Сделаны выводы об эффективности рассмотренных биомедицинских клеточных продуктов в лечении хронических ран, дана оценка проведенным исследованиям, даны рекомендации по наиболее эффективному использованию аллогенных дерматотропных биомедицинских клеточных продуктов

Реконструкция гортаноглотки с использованием аутологичных тканеинженерных эпителизированных лоскутов

After removal of metastatic malignant tumors of the hypopharynx and larynx, hypopharyngeal defects are formed. To restore the hypopharynx, a mucosa and a muscular component are needed.The objective of this study is to develop a hypopharyngeal reconstruction technique using prelaminated pectoralis major flap with mucosal epithelium analogue from autologous epithelial layers.Materials and methods. Nine patients underwent reconstruction of the hypopharynx using bioengineered prelaminated pectoralis major flaps. The mucosa was restored by tissue-engineered autologous epithelial cell layers that were obtained by culturing in vitro cells isolated from skin biopsies that were previously obtained from patients.Results. Oral nutrition was restored in all cases. Pharyngeal stenosis was detected in one (11%) patient. A stratified squamous epithelium on the pectoral fascia was revealed in 67% of cases at week 2 after prelamination, in 89% of cases at week 4 after reconstruction and in 100% of cases at month 3, 6, 12 and 24 after reconstruction.Conclusion. Reconstruction using prelaminated bioengineered flaps allows recreating the anatomical integrity and function of the hypopharynx.После удаления распространенных злокачественных опухолей гортаноглотки и гортани образуются дефекты верхних пищеварительных путей, для устранения которых необходимы слизистая оболочка и мышечный компонент.Целью данного исследования является разработка методики реконструкции гортаноглотки с использованием преламинированных пекторальных лоскутов с аналогом эпителия слизистой оболочки из аутологичных эпителиальных пластов.Материалы и методы. Девяти пациентам выполнена реконструкция гортаноглотки биоинженерными преламинированными пекторальными лоскутами с восстановлением слизистой оболочки тканеинженерными аутологичными эпителиальными клеточными пластами, которые были получены путем культивирования in vitro клеток, выделенных из предварительно полученных у пациентов биоптатов кожи.Результаты. Во всех случаях было восстановлено пероральное питание. У одного (11%) пациента был выявлен стеноз глотки. Многослойный плоский эпителий на фасции пекторального лоскута выявлен в 67% случаев через 2 недели после преламинации, в 89% случаев – через 4 недели после реконструкции и в 100% случаев – через 3, 6, 12, 24 месяца после реконструкции.Заключение. Реконструкция с использованием преламинированных биоинженерных лоскутов позволяет воссоздать анатомическую целостность и функцию гортаноглотки

Label-free sorting of iPS cells during neuronal differentiation using FLIM and multiphoton fluorescence microscopy

The changes in cell metabolism can affect the epigenome-modifying enzymes activity during iPSCs differentiation and thus control the functional potential of the final cell. Therefore, for therapeutic applications, the restoration of a fully functional mitochondrial network specific for the cell types derived from iPSCs will be required to support the energy and other mitochondrial factors. Recently, FLIM method allows to study the metabolic changes that accompanying cell differentiation noninvasively and without additional labels. In this study, we investigated the metabolic changes in iPSCs during neural differentiation using two-photon fluorescence microscopy and FLIM. Cellular metabolism was examined by monitoring the optical redox ratio (FAD/NAD(P)H), the fluorescence lifetime contributions of the free and bound forms of NADH and NADPH. Given that neural differentiation is also accompanied by synthetic proceßes and oxidative streß, this proceß was included in the scope of this work. We demonstrated an increased contribution of protein-bound NADH and NADPH in neuron aßociated with metabolic switch to oxidative phosphorylation and the biosynthetic proceßes or oxidative streß, respectively. We also found that the optical redox ratio FAD/NAD(P)H decreased during neural differentiation, and this was likely to be explained by the intensive lipid membrane synthesis or ROS generating and the enhanced NADPH production aßociated with them. The biochemical analysis was carried out to verify the metabolic status of iPSCs and their neural derivatives. Based on the data on glucose consumption, lactate and ATP amount we registered the trend to the metabolic pathways redistribution towards the oxidative phosphorylation in neuron. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only

Probing metabolic alteration of differentiating induced pluripotent stem cells using label-free FLIM

The differentiation of endothelial cells from human iPSC has incontestable advantages in diseases research and therapeutic applications. However, the safe use of iPSC derivatives in regenerative medicine requires an enhanced understanding and control of factors that optimize in vitro reprogramming and differentiation protocols. Shifts in cellular metabolism associated with intracellular pH changes affect the enzymes that control epigenetic configuration, which impact chromatin reorganization and gene expression changes during reprogramming and differentiation. FLIM-based metabolic imaging of NADH and FAD is a powerful tool for measuring mitochondrial metabolic state and widely used diagnostic method for identification of neoplastic diseases, skin diseases, ocular pathologies and stem cells differentiation. Therefore, in this study, we used the potential of FLIM-based metabolic imaging and fluorescence microscopy of NADH and FAD to study the metabolic changes during iPSC differentiation in endothelial cells. The evaluation of the intracellular pH was carried out with the fluorescent pH-sensor SypHer-2 and fluorescence microscopy to obtain complete information about metabolic status of iPSC and their endothelial derivatives. Based on the FAD/NAD(P)H optical redox ratios increase and the contributions rise of the NAD(P)H fluorescence lifetime in iPSC during endothelial differentiation, we demonstrated an contribution increase of OXPHOS to cellular metabolism. Based on the shift toward more acidic intracellular pH in endothelial cell derived from iPSCs we verified their oxidative state. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only

Tissue equivalent transplantation in the treatment of certain skin injuries

Chronic ulcers are a common and socially significant problem worldwide. Autodermoplasty is the gold standard treatment for chronic ulcers. However, it is not always possible to perform this surgical procedure for a rather large group of patients, due to some reasons, which include high risk of autodermotransplant rejection, lack of donor material, and patient’s unwillingness to undergo surgery with an often unpredictable result. A potential solution to the problem is to use skin equivalents from allogeneic donor material. The use of allogeneic (donor) human cells makes it possible to fill the deficit of the patient’s donor resources and close wound without causing additional injury to the patient. This paper provides an overview of the application of foreign and domestic biomedical cell products in clinical trials and real clinical practice. We draw conclusions on the efficiency of the considered biomedical cell products in the treatment of chronic ulcers, evaluate the conducted research, and make recommendations on the most efficient use of allogeneic dermatotropic biomedical cell products