[Closing of nasal septum perforation using adipose stromal vascular fraction: an experimental study]. / Zakrytie perforatsii peregorodki nosa s primeneniem stromal'no-vaskulyarnoi zhirovoi fraktsii: eksperimental'noe issledovanie.

Nasal septal perforation (NSP) is a complex problem in otorhinolaryngology, which leads to impaired nasal breathing and dryness in the nose. This reduces the patient's quality of life and leads to psychological discomfort. The treatment of nasal septum perforation is selected taking into account the clinical manifestations, perforation parameters and general condition of the patient. Currently, a large number of different surgical methods have been described in order to closing the defect of nasal septum. To date, there is no universally accepted method for closing NSP, which stimulates the search and development of new treatment options. OBJECTIVE: Under experimental conditions, to study a new method for closing nasal septum perforation using a collagen scaffold together with adipose stromal vascular fraction containing multipotent mesenchymal stromal cells. MATERIAL AND METHODS: The experiment was carried out on a model of nasal septum perforation in 24 male rabbits divided into four groups, depending on the construct, implanted into the defect zone: the 1st group was the control group - without the introduction of implantation material; the 2nd group - collagen scaffold without adipose stromal vascular fraction; the 3rd group - collagen scaffold with xenogenic adipose stromal vascular fraction; the 4th group - collagen scaffold with allogeneic adipose stromal vascular fraction with further dynamic evaluation of endoscopic control on day 14, after 1 month, 3 months, and 6 months. At month 6, the animals were removed from the experiment, followed by morphological examination in color with hematoxylin and eosin, as well as safranin and methyl green. RESULTS: As a result of the experiment using adipose stromal vascular fraction of allogeneic and xenogenic origin, closing of perforation of the nasal septum of a rabbit for 3 months of dynamic endoscopic control, as well as according to morphological research, was demonstrated. CONCLUSION: Our study showed that the use of adipose stromal vascular fraction containing not only endothelial cells and pericytes, but also multipotent mesenchymal stromal cells in combination with a collagen scaffold closes the perforation of the nasal septum in a rabbit, without increasing the risk of violations of habitual vital activity.

Biodegradable iron-silicon implants produced by additive manufacturing.

Due to many negative and undesirable side effects from the use of permanent implants, the development of temporary implants based on biocompatible and biodegradable materials is a promising area of modern medicine. In the presented study, we have investigated complex-shaped iron-silicon (Fe-Si) scaffolds that can be used as potential biodegradable framework structures for solid implants for bone grafting. Since iron and silicon are biocompatible materials, and their alloy should also have biocompatibility. It has been demonstrated that cells, mesenchymal stromal cells derived from the human umbilical cord (UC-MSC) and 3T3, were attached to, spread, and proliferated on the Fe-Si scaffolds' surface. Most of UC-MSC and 3T3 remained viable, only single dead cells were observed. According to the results of biological testing, the scaffolds have shown that deposition of calcium phosphate particles occurs on day one in the scaffold at the defect site that can be used as a primary marker of osteodifferentiation. These results demonstrate that the 3D-printed porous iron-silicon (Fe-Si) alloy scaffolds are promising structures for bone grafting and regeneration.

Regenerative Potential of a Suspension and Spheroids of Multipotent Mesenchymal Stromal Cells from Human Umbilical Cord on the Model of Myocardial Infarction in Rats.

Regenerative potential of multipotent mesenchymal stromal cells from the human umbilical cord (MMSC-UC) in the suspension and spheroid form was revealed during the progression of experimental small focal myocardial infarction in rats. In isoproterenol-induced myocardial infarction, foci of necrosis and inflammatory infiltrate and at later terms fibrosis foci were found mainly in the left ventricle of rat heart. In rats receiving MMSC-UC, destructive changes in the myocardium, fibrous scars, and inflammatory process were less pronounced. MMSC-UC also contributed to normalization of the morphofunctional parameters of the heart. Spheroids exhibited higher efficiency in comparison with cell suspension.

2D/3D buccal epithelial cell self-assembling as a tool for cell phenotype maintenance and fabrication of multilayered epithelial linings in vitro.

Maintaining the epithelial status of cells in vitro and fabrication of a multilayered epithelial lining is one of the key problems in the therapy using cell technologies. When cultured in a monolayer, epithelial cells change their phenotype from epithelial to epithelial-mesenchymal or mesenchymal that makes it difficult to obtain a sufficient number of cells in a 2D culture and to use them in tissue engineering. Here, using buccal epithelial cells from the oral mucosa, we developed a novel approach to recover and maintain the stable cell phenotype and form a multilayered epithelial lining in vitro via the 2D/3D cell self-assembling. Transitioning the cells from the monolayer to non-adhesive 3D culture conditions led to formation of self-assembling spheroids, with restoration of their epithelial characteristics after epithelial-mesenchymal transition. In 7 days, the cells within spheroids restored the apical-basal polarity, and the formation of both tight (ZO1) and adherent (E-cadherin) intercellular junctions was shown. Thus, culturing buccal epithelial cells in a 3D system allowed us to recover and durably maintain the morphological and functional characteristics of epithelial cells. The multilayered epithelial lining formation was achieved after placing spheroids for 7 days onto a hybrid matrix, which consisted of collagen layers and reinforcing poly (lactide-co-glycolide) fibers and was proven promising for replacement of the urothelium. Thus, we offer an effective technique of forming multilayered epithelial linings on carrier-matrices using cell spheroids that was not previously described elsewhere and can find a wide range of applications in tissue engineering, replacement surgery, and regenerative medicine.

Angiogenic potential of spheroids from umbilical cord and adipose-derived multipotent mesenchymal stromal cells within fibrin gel.

One of the essential goals in regenerative medicine is microvascularization which enables an effective blood supply within de novo constructed tissues and organs. In our study, we used two common multipotent mesenchymal stromal cell (MMSC) sources (subcutaneous adipose tissue and Wharton's jelly of the umbilical cord) where is a subpopulation of endothelial precursors. In the medium supplemented with VEGF, the 3D cultures of UC MMSCs and ADSCs promoted the endothelial cell differentiation. To evaluate their ability to form a capillary-like network, we encapsulated spheroids within non-modified and PEGylated fibrin hydrogels. The PEGylated hydrogel supported better the formation of multibranched cords than the pure fibrin gel. Analysis of tubule growth rate, length, and branching showed that the differentiated ADSCs had higher angiogenic potential than the differentiated hUC MMSCs. Our study can be a basis for the development of new strategies in tissue engineering and treatment of vascular diseases.

[Cellular Model Based on Laser Microsurgery of Cell Spheroids to Study the Repair Process].

In this study, modern techniques of laser microsurgery of cell spheroids have been used to develop a new simple, reproducible model for studying the mechanisms of repair and regeneration in vitro. Nanosecond laser pulses were applied to perform a microdissection of the outer and the inner zones of the spheroids from dermal fibroblasts. To achieve effective dissection and preservation of spheroid viability, the optimal parameters were chosen: 355 nm wavelength, 100 Hz frequency, 2 ns pulse duration, laser pulses in the range of 7­9 µ J. After microdissection, we observed injury of the spheroids : the edges of the wound surface opened and the angular opening reached a value of more than 180°. As early as during the first hour after spheroid microdissection with laser radiation, the surviving cells changed their shape: cells on the spheroid surface and directly in the damaged area became rounded. One day after microdissection, the structure of the spheroids began to partially recover, the cells in the surface layers began to take the original flattened shape; debris of dead damaged cells and their fragments was gradually cleared from the spheroid composition. In the proposed model, the first data on stimulation of structure recovery of injured spheroids from dermal fibroblasts with a P199 synthetic polypeptide, which is used in cosmetology for the initiation of antiaging and regenerative effects in the skin, were received. After microdissection, recovery of the spheroids structure with a few surface layers of flattened imbricated arranged cells and polygonal cells of the inner zone in the presence of P199 peptide was faster than in the control group, and was completed within 7 days, presumably due to the remodeling of the survived cells.

The technology of obtaining multipotent spheroids from limbal mesenchymal stromal cells for reparation of injured eye tissues.

It is known that stem and progenitor cells open new possibilities for restoring injured eye tissues. Limbal eye zone, formed mainly by derivatives of neural crest, is the main source of stem cells for regeneration. The current study considers development of innovative technology for obtaining 3D spheroids from L-MMSC. It was shown that under 3D conditions L-MMSC due to compactization and mesenchymal-epithelial transition self-organize into cellular reparative modules. Formed L-MMSC spheroids retain and promote undifferentiated population of stem and progenitor limbal cells, as supported by expression of pluripotency markers - Oct4, Sox2, Nanog. Extracellular matrix synthetized by cells in spheroids allows retaining the functional potential of L-MMSC that are involved in regeneration of both anterior and, probably, posterior eye segment.

3D-technology of the formation and maintenance of single dormant microspheres from 2000 human somatic cells and their reactivation in vitro.

We developed an original reproducible 3D-technology for preparation of single dormant microspheres consisting of 2000 somatic cells. The dynamics of microsphere assembly from mesenchymal and epithelial cells of retinal pigment epithelium was traced using time-lapse microscopy: formation of a loose aggregate over 24 h followed by its gradual consolidation and formation of a compact viable microsphere with a diameter of 100-150 µ by day 7. The cell number in the formed microspheres remains unchanged. Reactivation observed upon fusion of epithelial and/or mesenchymal microspheres results in the formation of a united compact microtissue. The fusion dynamics reproduces spherogenesis irrespective of the initial amount of co-cultured microspheres. Reactivation via two-step induced angiogenesis opens new prospects for production of vascularized microspheres and microtissues.

[Experimental study of safety in application of the Iris Medical IQ 810 diode laser in clinical treatment of age-related macular degeneration].

During the study of diode laser radiation effect in micropulse mode on culture cells of human retinal pigment epithelium it was revealed that the quota of dead cells was a minimum. Besides, a certain conformity between dead cells quota and parameter characteristics of laser radiation. Based on the performed experimental study it was revealed that for a work using the Iris Medical IQ 810 diode laser in the micropulse mode following parameters: duration of pulse set--300ms, duration of function--9.1%, power--750mW are safe for retinal pigment epithelium cells. Rationales of safety in application of the infrared diode laser radiation in micropulse mode in clinic for treatment of age-related macular degeneration (AMD) exampled by cell culture of human retinal pigment epithelium.

Behavior of Transplanted Multipotent Cells after in Vitro Transplantation into the Damaged Retina.

The use of stem cell technologies in retinal defect reparation therapy has produced beneficial results. Nowadays, numerous protocols exist which provide a neural differentiation of the stem cells transplanted into the retina. However, questions concerning the functional replacement of the missing retinal neurons by transplanted cells thus far remain unanswered. The organotypic culture protocol was used in this study in order to prove the possibility of transdifferentiation of bone marrow stromal cells (MMSCs) and neural stem/progenitor cells (NSPCs) from EGFP-positive mice and the functional integration of these cells. This technique enables a detailed characterization of cell behavior post-transplantation. Using atomic force microscopy, we reliably demonstrated the difference (p < 0.01) between the thickness of the outgrowths formed by glial and endothelial retina cells and the thickness of neurites and neuro-like transplanted MMSC outgrowths. MMSCs are also shown to form synapses up to 2.5 ± 0.06 µm in diameter on day 4 after the transplantation. Following electrical stimulation (20V, 0.5Hz, 200ms), clear depolarization of retinal neurons and their outgrowths is detected. It is shown that some of these GFP+ MMSCs, which changed their morphology after the transplantation in retinal explants to neuro-like MMSCs, are capable of depolarizing after exogenous stimulation.

[Ultrasonographic guidance in diagnostic and therapeutic interventions in diseases of the internal organs]. / Ul'trazvukovoi metod kontrolia za diagnosticheskimi i lechebnymi vmeshatel'stvami pri zabolevaniiakh vnutrennikh organov.

A hundred fifty four invasive diagnostic and therapeutical interventions were made in patients with diseases of the abdomen and retroperitoneal space under ultrasonographic guidance. Various biopsies were performed in 139 patients; positive results were achieved in 120 (86.3%) cases. In 15 patients, diagnostic biopsies were combined with therapeutical interventions, such as aspiration of cysts in the liver (n = 3) and kidney (n = 2); drainage of abscesses in the abdomen (n = 5) and liver (n = 5). Fourteen patients with mechanical jaundice caused by extrahepatic bile duct tumors or pancreatic head cancer underwent percutaneous transhepatic cholecystocholangiography followed by external drainage. Percutaneous transhepatic drainage of the gallbladder was made in 1 patient with acute cholecystitis.

[MRI diagnosis of soft tissue tumors]. / MR-tomograficheskaia diagnostika opukholei miagkikh tkanei.

MRI findings of 13 patients with soft tissue tumors (STT) are presented. There are were abnormalities, such as primary STTs of the hip in 5 patients, the back in 1, and the neck in 1, STT relapses of the hip in 2 and those of the back in 1. Two patients had chronic hip STT hematomas and 1 had hip STT metastatic melanoma. The diagnosis was verified in 11 cases (in 10 cases at surgery and 1 at needle biopsy). MRI makes it possible to define the accurate sizes of the tumor, its structure and relationship to its adjacent tissues, which is important in choosing at treatment policy, the type and scope of a surgical interventions.

[Puncture methods of the diagnosis and treatment under the control of US, CT and X-ray video image]. / Punktsionnye metody diagnostiki i lecheniia pod kontrolem UZ, KT i rentgenotelevizionnogo izobrazheniia.

Exploratory and therapeutical punctures were made in 454 patients under the control of ultrasound, CT, and X-ray teleimage. Diagnostic biopsies established the cytological and histological nature of diseases in 67.6% of cases, positive ultrasound-, CT-, and X-ray teleimage-controlled biopsies being 76.3, 64.9, and 71.4%, respectively. Beneficial effects of therapeutical interventions were obtained in 89.5% of patients.

[Diagnostic possibilities of CT-guided lung biopsy]. / Diagnosticheskie vozmozhnosti biopsii legkikh pod kontrolem KT.

The authors analyse the results of computer-aided tomographic biopsies in 81 patients with various pulmonary diseases (63 with tumors, 53 of these with malignant ones, and 18 with nontumors diseases). Positive results were obtained in 86.4% of cases. Biopsies proved to be informative in 44 out of 48 patients with lung cancer, in 8 out of 10 ones with benign tumors, and in 10 out of 12 ones with nonspecific inflammations. The results were not informative in 13.6% of patients. In 4 cases the failure was caused by the absence of cellular material in the puncture biopsy specimen, in 5 it was due to the presence of only pulmonary cells. In two more cases the diagnosis of lung cancer made by computer-aided biopsy was erroneous because of the presence of inflammation elements and amorphous mass in the examined. Complications occurred in 22 (27.2%) patients, pneumothorax in 16 (19.8%) and local pulmonary hemorrhages in 6 (7.4%) of these. Hence, computer-aided biopsy of the lung is an informative diagnostic method, permitting a correct diagnosis in the majority of cases.

[Computerized myelotomography in lesions of the spine and spinal cord]. / Komp'iuternaia mielotomografiia pri porazheniiakh pozvonochnika i spinnogo mozga.

CT myelotomography was carried out in 262 patients with various conditions of the spine and cord. Pathologic changes were detected in 233 (87.6%) of the examinees. The most frequent diseases were tumors and metastases (59 cases), degenerative processes, including primary and recurrent hernias (59 cases), atrophic processes (46 cases). The accuracy of the diagnosis was found to be 98.6%. A false-positive result was obtained in one case with a suspected tumor of the cord. No complications were recorded. Computer-aided myelotomography is an effective method for the diagnosis of involvement of the spine and cord.

[Errors and complications in computed tomography-guided biopsies]. / Oshibki i oslozhneniia pri biopsiiakh pod kontrolem komp'iuternoi tomografii.

The article analyses the results of 104 biopsies which were carried out in 84 patients with different diseases of the organs of thoracic (50) or abdominal (16) cavities, postperitoneal space (32), bones and soft tissues (6). Positive results were obtained in 67 cases, negative--in 26, false-negative--in 11. The specificity was 70.3%, sensitivity--100%, authenticity--64.4%. The authors make a conclusion that the results and authenticity of biopsies depend on such factors as the proper organ to be punctured, character and structure of the affection, its location, volume and spreading, as well as the type of needles to be used.

[Invasive interventions with computerized tomography guidance]. / Invazivnye vmeshatelstva pod kontrolem kompíuternoi tomografii.

Invasive interventions (diagnostic and therapeutic) were conducted on 125 patients. Fifty-six has diseases of the lungs, 16 of the mediastinum, 4 of the pleura, 17 of the liver, 2 of the pancreas, one of the spleen, 15 of the kidneys, 7 of the retroperineal space, two of the soft tissues, and 5 patients had diseases of the skeletal bones. Biopsy was combined with aspiration of the renal cyst content in 7 cases. The diagnostic value of biopsy under control of CT was 86% (106 cases); negative and pseudonegative results were recorded in 18 (14%) patients. The efficacy of biopsy is determined by such factors as the size of the lesion and the depth of its localization in the involved organ, the character of the pathological process and the region of its localization, the site from which the material is collected, and the needles used. Among 7 patients with therapeutic interventions, 5 had good and 2 had satisfactory immediate results. Complications were revealed in 4 (3.3%) patients, pneumothorax was encountered most frequently.

[The potentials of computed tomography in the treatment of cysts and inflammatory diseases of the internal organs]. / Vozmozhnosti komp'iuternoi tomografii v lechenii kist i vospalitel'nykh zabolevanii vnutrennikh organov.

Computed tomography (CT) is used more and more often recently in the treatment of some surgical diseases of the internal organs. The subject of discussion in this work are the results of treatment of cysts and pyoinflammatory diseases of the internal organs. The work was conducted on CT IV generation "QUAD 1" of the CMS firm (England). Needles measuring 14-18d in diameter (Charrière scale) were used for aspiration. A total of 23 therapeutic interventions were performed in 19 patients with exudative pleurisy (1 patient), pleuritis (4), pulmonary abscesses (1), abscesses of the liver (1), abscesses of the subhepatic and subdiaphragmatic spaces (2), cysts of the liver (1) and kidneys (1 patient). Among the interventions 21 were aspirations and 2 drainage of the abscess. A mechanical syringe is used lately for more effective evacuation of the contents. The removed contents, ranging in volume from 10.0 to 600.0 ml, was subjected to cytological, biochemical, and bacteriological analysis. A single puncture is sufficient, as a rule, for aspiration of the cyst contents, one-stage aspiration from cysts of both kidneys may be conducted (as was the case in two of our patients). 96% alcohol was used as the sclerosing agent. In cases of small abscesses aspiration may be repeated for removal of the contents, irrigation of the cavity, and administration of antibiotics. In abscesses measuring more than 10 cm drainage is advisable. Control CT examination is conducted once a week. The condition of the cavity is appraised by combining CT with its contrast study. The immediate results were good and satisfactory in 17 (94.7%) patients.(ABSTRACT TRUNCATED AT 250 WORDS)