Correlative Fluorescent Scanning Probe Nanotomography Used to Study the Intracellular Distribution of Doxorubicin in MCF-7 Human Breast Adenocarcinoma Cells.

Developing technologies for efficient targeted drug delivery for oncotherapy requires new methods to analyze the features of micro- and nanoscale distributions of antitumor drugs in cells and tissues. A new approach to three-dimensional analysis of the intracellular distribution of cytostatics was developed using fluorescence scanning optical-probe nanotomography. A correlative analysis of the nanostructure and distribution of injected doxorubicin in MCF-7 human breast adenocarcinoma cells revealed the features of drug penetration and accumulation in the cell. The technology is based on the principles of scanning optical probe nanotomography and is applicable to studying the distribution patterns of various fluorescent or fluorescence-labelled substances in cells and tissues.

Stem and Somatic Cell Monotherapy for the Treatment of Diabetic Foot Ulcers: Review of Clinical Studies and Mechanisms of Action.

Diabetic foot ulcer (DFU) is one of the most severe complications of diabetes mellitus, often resulting in a limb amputation. A cell-based therapy is a highly promising approach for an effective DFU treatment. However, there is no consensus regarding the most effective cell type for DFU treatment. Various cell types contribute to chronic wound healing via different mechanisms. For example, application of keratinocytes can stimulate migration of native keratinocytes from the wound edge, while mesenchymal stem cells can correct limb ischemia. To assess the effectiveness of a certain cell type, it should be administered as a monotherapy without other substances and procedures that have additional therapeutic effects. In the present review, we described therapeutic effects of various cells and provided an overview of clinical studies in which stem and somatic cell-based therapy was administered as a monotherapy. Topical application of somatic cells contributes to DFU healing only, while injection of mesenchymal stem cells and mononuclear cells can break a pathophysiological chain leading from insufficient blood supply to DFU development. At the same time, the systemic use of mesenchymal stem cells carries greater risks. Undoubtedly, cell therapy is a potent tool for the treatment of DFU. However, it is vital to conduct further high-quality clinical research to determine the most effective cell type, dosage and way of administration for DFU treatment. Ischemia, neuropathy and neuro-ischemia are underlying factors of diabetic foot ulcer. Stem and somatic cells monotherapy can improve chronic wound healing via different mechanisms.

Biological Activity of Agaricinic Acid Nanoparticles against Human Hepatoma HepG2 Cells.

A stable preparation of agaricinic acid nanoparticles was obtained. The mean hydrodynamic size of nanoparticles according to photon correlation spectroscopy was 200 nm and zeta potential was -57 mV. Cytotoxic activity of agaricinic acid nanoparticles against human HepG2 hepatoma cells was evaluated. Nanoparticles with a low concentration of agaricinic acid stimulated and with high concentration - suppressed metabolic activity and viability of hepatoma cells. The EC50 for the stimulating effect was 32.8 µg/ml, and the IC50=602.1 mg/ml. The preparation of agaricinic acid nanoparticles can be used in medicine as a potential antitumor agent.

[Neurohumoral response and Fas-ligand-induced apoptosis in peripheral blood of patients with acute ischemic stroke]. / Neirogumoral'nyi otvet i aktivnost' Fas-liganda perifericheskoi krovi v ostrom periode ishemicheskogo insul'ta.

OBJECTIVE: To identify an effect of the neurohumoral response on the severity and orientation of Fas-ligand-initiated processes in the acute period of IS. MATERIAL AND METHODS: The study included 155 patients with IS in the territory of the left and right middle cerebral arteries, the control group consisted of 28 people. The National Institutes of Health Stroke Scale (NIHSS), the Hospital Anxiety and Depression Scale (HADS), and the Daily Life Stress scale were used. Concentrations of sFas, sFasL, cortisol (K), adrenaline (A), norepinephrine (NE), adrenocorticotropic hormone (ACTH) in the blood plasma of patients with IS were measured by enzyme-linked immunosorbent assay on days 1, 7 and 21 and once in the control group. CD3CD95+ lymphocytes phenotyping was performed using flow cytometry. RESULTS AND CONCLUSION: The hypothalamic-pituitary-adrenal axis dominance is associated with the activation of the apoptosis-inducing properties of peripheral blood in the first week after the IS onset and their decrease towards the end of the acute period, which is clinically represented by the increased levels of anxiety and depression, an unfavorable outcome of the acute period of IS.

[Long-term oucomes of tracheal transplantation: success and unsolved problems]. / Otdalennyi rezul'tat transplantatsii trakhei: uspekh i nereshennye problemy.

AIM: To analyze long-term outcomes of tracheal transplantation. MATERIAL AND METHODS: There were 1128 patients with cicatricial tracheal stenosis who have been operated at the Petrovsky Russian Research Center for Surgery and the Sechenov First Moscow State Medical University for the period 1963-2015. RESULTS: Operations have become safer. Postoperative morbidity and mortality reduced from 41.4% (1963-1980) to 5.6% (2001-2015) and from 21.9% (1963-1980) to 0.5% (2001-2015), respectively. Tracheal transplantation was performed in 2 cases and fundamentally different tracheal structures were applied. Donor thyreotracheal complex with restored blood supply through thyroid vessels was used in the first case (2006). Perennial experimental trials preceded clinical application of this technique. In the second case (2010) we applied scientific results of foreign colleagues (cellular technologies and methods of regenerative medicine to create artificial trachea). Patients are still alive after 12 and 8 years, respectively. Restoration of blood supply of donor trachea is possible through thyroid collaterals. This technique is successful in long-term period. Tissue-engineered trachea cannot be considered true trachea due to no all tracheal components. However, such trachea provides air-conducting, evacuation and protective functions. Tracheomalacia requires further researches as one of the main problems of tracheal transplantation.

[Apoptosis as a systemic adaptive mechanism in ischemic stroke]. / Apoptoz kak sistemnyi adaptivnyi mekhanizm pri ishemicheskom insul'te.

This paper presents a literature review considering the role and mechanism of apoptosis in the pathogenesis of ischemic stroke (IS). The authors introduce a new concept: the functional request of the patient as a set of external (the nature and intensity of rehabilitation measures, characteristics of everyday life, diet, etc.) and internal (genetic factors, internal picture of the disease, availability of rental and other psychological facilities and etc.) attributes. This concept allows a new angle in understanding the pathogenesis of IS and creates fundamental and clinical potential for more successful approaches to therapy and rehabilitation after IS.

[Possible use of the growing liver biological set for hepatic recovery after toxic damage (an experimental study)]. / Vozmozhnosti biologicheskoi kombinatsii, poluchennoi iz rastushchei pecheni, dlia ee vosstanovleniia pri toksicheskom povrezhdenii (éksperimental'noe issledovanie).

The lack of acceptable pharmacological approaches for restoration of the injured liver is associated with complex of mechanisms involved in hepatic regeneration and with difficulty of the target selection. The aim of this research was to study the hepatoprotective function of the extract from both the growing and regenerating liver containing a natural set of factors crucial for the hepatic restoration. Extracts from both regenerating liver of rats after 70% hepatic resection and the growing liver of neonatal pigs were obtained using own original technique. The set of resultant extracts was named as the hepatic regeneration set (HRS). HRS fractionation was carried out using the Toyopearl HW-50S sorbent. The efficiency of HRS and its fractions was estimated using a model of the mouse liver thioacetamide injury and monitoring hepatic enzyme activity in blood serum. The activities of AST and ALT in intact animals were 50 U/l and 80 U/l, respectively; after thioacetamide administration they increased to 2059±212 U/l and 4280±440 E/l, respectively (p<0.05). Treatment of injured animals with HRS from the rat regenerating liver resulted in a significant decrease of transaminase activities to 924±148 U/l (AST; p<0.05) and 1633±308 U/l (ALT; p<0.05). A similar effect was observed after treatment with HRS from the neonatal pig liver: the AST decreased to 937±138 U/l (p<0.05), while ALT activity decreased to 1710±237 U/l (p<0.05). HRs fractionation resulted in identification two active fractions characterized by much higher (8-29) hepatotropic effect that that of the whole extract. These fractions contained peptide/protein components with the range of molecular mass of 3-60 kDa (fraction 1) and 3-25 kDa (fraction 2a). Fraction 1 also contained some polynucleotides in fraction 1. Subsequent studies of these fractions exceeding the hepatotropic effect of original HRS is clearly needed to identify their individual components by immunochromatography methods, ELISA, MRM mass spectrometry and quantitative PCR.

[Urethra Reconstruction with Tissue-Engineering Technology].

Urethral stricture is a disease characterized by a pathological narrowing of the urethra. Treatment for this condition often requires surgery using autologous grafts (urethroplasty). It is common practice to use patient's own tissue like genital and extragenital skin, tunica vaginalis, buccal mucosa as a source of the graft. Alternative and safer approach is to use tissue-engineered graft created in a laboratory using patient's autologous cells and biocompatible matrix (scaffold). The article presents the up-to-date achievements in lab-created tissue-engineered graft, describes all components needed to build a tissue-engineered structure of the graft for urethroplasty, and summarizes authors' thoughts on advantages and disadvantages of various approaches to choose both cellular component and the matrix of future construction. The article reviews clinical studies conducted in the field of tissue engineering of the graft material for urethraplasty.

[The Possibilities of Cell Technologies in the Treatment of Cicatricial Lesions of the Vocal Folds].

The article is a brief review of publications devoted to the problem of persistent dysphonia. The main cause of voice disorders is the scarring of the vocal folds resulting from trauma, surgical manipulation, inflammatory process. Treatment of cicatricial lesions of the vocal folds remains a challenge, as far as existing methods do not ensure the recovery of the ultrastructure of the vocal folds. The authors present modern data on the structure of the vocal folds at the cellular level. Considered pathologic processes occur in different stages of scarring. Applied technologies of phonosurgery and conservative treatment, their effectiveness and shortcomings are covered. Analysis of experimental research conducted in the world demonstrates the promise of using the methods of tissue engineering to treat scarring of the vocal folds and to restore the microstructure of the latter. Identified current issues remain unresolved, which leads to the need for further experimental and clinical studies in the treatment of this pathology.

IMMUNOMODULATORY EFFECT OF ALLOGENEIC MESENCHYMAL STEM CELLS OF RATS.

INTRODUCTION: Recently, more and more attracted the attention of cell therapy, which requires a study of the efficacy and safety of allogeneic MSCs transplantation in acute and chronic inflammatory reactions. The aim of our study was to examine the effectiveness of transplantation of allogeneic mesenchymal bone marrow stromal cells for the healing of surgical wounds the glandular stomach in rats. MATERIAL AND METHODS: Using white Wistar rats. Producing cell transplantation mononuclear fraction derived from rat bone marrow aspirate. Injected cells 8 and 9 th passage. The dose of cells administered to 3-th and / or 7-th days 3,5h106 cells / ml twice or 5,0h106 cells / ml dose. Autopsy on day 10-th and 17-th. The serum ELISA determined the content of the pro- and anti-inflammatory cytokines IL1ß, TNFα, IFNy, IL-4. RESULTS AND DISCUSSION: Introduction MSCs contributed to the decline of the immune mediators of inflammation IL1P, TNFa, IFNy, increase anti-inflammatory IL4. Quality improved healing. CONCLUSION: Rapid curative effect of stem cells may be associated with the formation of blood immune cells (macrophages) that produce substances that restore damaged tissue. They restore the balance between Th1 and Th2.

[PLGA mesh-collagen hybrid scaffold and tissue-engineered product in substitution urethroplasty: experimental validation].

Urethral strictures are a pressing issue in modern medicine. Substitution urethroplasty is considered one of the most effective treatment methods. However, despite the surgery showing good results, many problems remain unresolved, one being substitute material deficiency in extensive or recurrent strictures, as well as in cases requiring multistage surgeries, including those used to treat hypospadias. Graft removal also leaves the donor area prone to diseases and increases the length of surgery leading to a higher risk of intra- and postoperative complications. Tissue engineering (namely tissue-engineered products comprised of scaffolds and cells) may be a useful tool in dealing with these issues. The authors assessed the characteristics of a novel hybrid scaffold created from "reconstructed" collagen and a poly(lactic-co-glycolic acid) mesh. The resulting composite product showed good mechanical properties and functional performance. The hybrid scaffold was non-cytotoxic and provided an adequate base for cell adhesion and proliferation. Biodegradation resulted in the scaffold being replaced by urothelium and urethral mucosa. The newly formed tissues possessed adequate structural and functional properties. Only one rabbit out of 12 developed urethral stricture at the site of scaffold implantation. The above-mentioned facts suggest that the novel hybrid scaffold is a promising tissue-engineered product with potential implication in substitution urethroplasty.

[In vitro Cultivation of Functioning Passaged Ciliated Epithelium for Trachea Tissue Engineering].

BACKGROUND: Currently all tissue engineered trachea transplants had no ciliated epithelium until transplantation, and long-term temporary lack of mucociliary clearance leads to patients' condition decline and reduced life quality in postoperative period. So, the need for a better cultivation method and studying ciliated epithelium growth characteristics in cell culture increased rapidly. OBJECTIVE: The aim of our study was to investigate cultivation offunctionally complete passaged ciliated epitheliumfor trachea tissue engineering. METHODS: Human ciliated epithelium isolated from intraoperative bioptate was used for culturing in the special complex medium with morphological and functional characteristics evaluation. RESULTS: Ciliated epithelial cell-groups were obtained by culturing in the special complex medium. Generated cell-groups had ciliary activity and showed well-coordinated movement with functional characteristics similar to native epithelial tissue. The basic parameters of cell-activity were studied. CONCLUSION: Thus our study provides a new insight for the problem of ciliated epithelium in vitro culturing as well as developing the optimal laboratory method.

[Pathophysiological aspects of wound healing in normal and diabetic foot].

The main cause of long-term healing of ulcers in patients with diabetic foot is considered to be direct mechanical damage when walking due to reduced sensitivity to due to neuropathy, hyperglycemia, infection and peripheral artery disease. These factors determine the standard approaches to the treatment of diabeticfoot, which include: offloading, glycemic control, debridement of ulcers, antibiotic therapy and revascularization. Recently, however, disturbances in the healing process of the skin in diabetes recognized an additional factor affecting the timing of healing patients with diabetic foot. Improved understanding and correction of cellular, molecular and biochemical abnormalities in chronic wound in combination with standard of care for affords new ground for solving the problem of ulcer healing in diabetes.