[Texture analysis of 3D models for the prediction of the grade of clear cell renal cell carcinoma of the kidney (pilot study)].

AIM: To evaluate the possibilities of textural analysis of 3D models in differentiating the degree of nuclear dysplasia of the clear cell renal cell carcinoma (ccRCC). MATERIALS AND METHODS: The specimens after surgical treatment of 190 patients with ccRCC were analyzed. In all cases, nephron-sparing surgery (NSS) was performed through laparoscopic access. The clinical characteristics were evaluated, including age, gender, tumor localization (side, surface and segments), absolute tumor volume, Charlson comorbidity index, body mass index, nephrometry scores (RENAL, PADOVA, C-index). Patients were divided into 2 groups. In group 1, there were 119 patients with the ccRCC of Grade 1 or 2, while group 2 consisted of 71 patients with ccRCC of Grade 3 and 4. All patients underwent 3D virtual planning of procedure using the 3D modeling program "Amira". At the first stage, two experienced radiologists performed manual segmentation of 3D models of kidney parenchyma tumors. At the second stage, the tumor shape was analyzed with a mathematical calculation of three indicators and more than 300 textural features of statistics of types 1-2 were extracted. Further, an intellectual analysis was carried out. For the evaluation of tumor grade according to Furman system, the classification problem was solved using the machine learning algorithm Stochastic Gradient Descent and cross-validation k=5. RESULTS: The accuracy of classification for the two groups of Grade 1 or 2 and Grade 3 or 4 on the F1 metric was 72.2. To build the model, the following parameters were selected: the absolute tumor volume, the Charlson comorbidity index, "Energy", the first quartile and the second decile of the pixel intensity distribution. CONCLUSION: The texture analysis of 3D models for the prediction of Fuhrman grade in ccRCC demonstrated satisfactory quality for two groups of Grade 1 or 2 and Grade 3 or 4 nuclear dysplasia.

[Multidisciplinary approach in urology. Research aspect].

Multidisciplinary studies in urology is becoming a new global trend which is associated with an emerging of omix technology, big data, the development of IT and other technologies. The concept of "multidisciplinary approach" as well as a brief historical analysis of the development of urologic science are discussed in the article, based on the international Scopus database. An example of a multidisciplinary approach implemented in the Science and Technology Park for Biomedicine of Sechenov University is given. The stages of the creation and implantation of the tissue-engineering urethral wall are described. For the successful development of urologic researches, the active involvement of specialists from various fields of knowledge is required. Such a combination of competencies has already allowed to solve problems of an extremely high level of complexity now, and not in the distant future.

[Use of local baroimpulse therapy in the treatment of patients with chronic abacterial prostatitis / chronic pelvic pain syndrome and erectile dysfunction].

AIM: To study the efficiency of local baroimpulse therapy (BIT) in the complex treatment of patients with chronic abacterial prostatitis/chronic pelvic pain syndrome (CP/CPPS) and erectile dysfunction (ED). MATERIALS AND METHODS: A total of 112 men with confirmed diagnosis of CP/CPPS and ED, who received drug therapy according to the standards and guidelines were included in the study. In the main group (n=68) all patients additionally received local BIT in the form of rectal pneumovibromassage (PVM) of the prostate (totally, 10 procedures). In the control group (n=44) only standard treatment was provided. The efficiency of therapy was assessed based on the complex clinical examination, laboratory tests and imaging methods. RESULTS: At the visit 2 (1 month after the start of therapy) there were significant differences in clinical (resolution of symptoms of CP/CPPS and improvement of erectile function), laboratory and imaging criteria between 2 groups. The rectal PVM using apparatus MKV-01 "Inavita" resulted in decrease the activity of inflammation process and levels of pro-inflammatory cytokines, increase of flow rate and intraprostatic blood flow. According to the analysis of long-term results, after 6 months of therapy the remission was seen in the most of patients in the main group (79.4%), which was 40.8% higher than in the control group. CONCLUSION: The use of local BIT by mean of rectal PVM allows to increase efficiency of complex treatment of patients with CP/CPPS and ED.

[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.

[Idiopathic hypercalciuria. Diagnosis and treatment].

Most patients with idiopathic hypercalciuria and calcium nephrolithiasis have a family history of the disease. Idiopathic hypercalciuria is a metabolic abnormality with various causes and developmental pathways. The systematic review describes specific mutations associated with idiopathic hypercalciuria and nephrolithiasis. Detection of these mutations may provide a better understanding of the pathogenesis of this heterogeneous disease and personalize patient management depending on the detected polymorphisms. A promising treatment option for a mutation in the vitamin D receptor gene is thiazide diuretics in combination with bisphosphonates. Among bisphosphonates, the drug of choice which has been most strongly supported by research evidence is alendronate.

[Using of cell biocomposite material in tissue engineering of the urinary bladder].

In a systematic review, to present an overview of the current situation in the field of tissue engineering of urinary bladder related to the use of cell lines pre-cultured on matrices. The selection of eligible publications was conducted according to the method described in the article Glybochko P.V. et al. "Tissue engineering of urinary bladder using acellular matrix." At the final stage, studies investigating the application of matrices with human and animal cell lines were analyzed. Contemporary approaches to using cell-based tissue engineering of the bladder were analyzed, including the formation of 3D structures from several types of cells, cell layers and genetic modification of injected cells. The most commonly used cell lines are urothelial cells, mesenchymal stem cells and fibroblasts. The safety and efficacy of any types of composite cell structures used in the cell-based bladder tissue engineering has not been proven sufficiently to warrant clinical studies of their usefulness. The results of cystoplasty of rat bladder are almost impossible to extrapolate to humans; besides, it is difficult to predict possible side effects. For the transition to clinical trials, additional studies on relevant animal models are needed.

[Tissue engineering of urinary bladder using acellular matrix].

INTRODUCTION: Tissue engineering has become a new promising strategy for repairing damaged organs of the urinary system, including the bladder. The basic idea of tissue engineering is to integrate cellular technology and advanced bio-compatible materials to replace or repair tissues and organs. AIM: of the study is the objective reflection of the current trends and advances in tissue engineering of the bladder using acellular matrix through a systematic search of preclinical and clinical studies of interest. MATERIALS AND METHODS: Relevant studies, including those on methods of tissue engineering of urinary bladder, was retrieved from multiple databases, including Scopus, Web of Science, PubMed, Embase. The reference lists of the retrieved review articles were analyzed for the presence of the missing relevant publications. In addition, a manual search for registered clinical trials was conducted in clinicaltrials.gov. RESULTS AND DISCUSSION: Following the above search strategy, a total of 77 eligible studies were selected for further analysis. Studies differed in the types of animal models, supporting structures, cells and growth factors. Among those, studies using cell-free matrix were selected for a more detailed analysis. CONCLUSION: Partial restoration of urothelium layer was observed in most studies where acellular grafts were used for cystoplasty, but no the growth of the muscle layer was observed. This is the main reason why cellular structures are more commonly used in clinical practice.

[Using cell technologies to treat urologic diseases].

Stem and progenitor cells being introduced into the body have the ability to stimulate regeneration of tissues and organs by differentiating into specialized cells. Stem cell therapy is used in urology to treat various disorders, including erectile dysfunction, urinary incontinence, Peyronies disease, and male infertility. This review presents the results of international preclinical and clinical research on stem cell based medications for treating the above diseases. The most promising appears to be the use of adipose tissue-derived mesenchymal stem cells.

[Morphology of collagen matrices for tissue engineering (biocompatibility, biodegradation, tissue response)].

OBJECTIVE: to perform a comparative morphological study of biocompatibility, biodegradation, and tissue response to implantation of collagen matrices (scaffolds) for tissue engineering in urology and other areas of medicine. MATERIAL AND METHODS: Nine matrix types, such as porous materials reconstructed from collagen solution; a collagen sponge-vicryl mesh composite; decellularized and freeze-dried bovine, equine, and fish dermis; small intestinal submucosa, decellularized bovine dura mater; and decellularized human femoral artery, were implanted subcutaneously in 225 rats. The tissues at the implantation site were investigated for a period of 5 to 90 days. Classical histology and nonlinear optical microscopy (NLOM) were applied. RESULTS: The investigations showed no rejection of all the collagen materials. The period of matrix bioresorption varied from 10 days for collagen sponges to 2 months for decellularized and freeze-dried vessels and vicryl meshes. Collagen was prone to macrophage resorption and enzymatic lysis, being replaced by granulation tissue and then fibrous tissue, followed by its involution. NLOM allowed the investigators to study the number, density, interposition, and spatial organization of collagen structures in the matrices and adjacent tissues, and their change over time during implantation. CONCLUSION: The performed investigation could recommend three matrices: hybrid collagen/vicryl composite; decellularized bovine dermis; and decellularized porcine small intestinal submucosa, which are most adequate for tissue engineering in urology. These and other collagen matrices may be used in different areas of regenerative medicine.

[Kinetics of methyluracil release from bioresorbable polymeric carriers].

Bioresorbable poly(lactic-co-glycolic) matrix-carriers containing 20 wt. % of 6-methyluracil (MU) have been prepared by supercritical fluid monolithization without organic solvents. Raman spectroscopy was used to analyze both the spatial distribution MU over polymer matrices and the MU release kinetics from the carrier into phosphate buffer solution. It was found that, during the first 24 h, the amount of released MU did not exceed 15-20% of its encapsulated content. After that, the MU release kinetics passed to almost linear regime with simultaneous retarding of the process. On the 40th day of observation, the MU content in solution reached up to 80% of its initial content in the carriers. Thus, using 6-methyluracil as a model, it was shown that the proposed bioresorbable and bioactive composites can be used as matrix-carriers for targeted and long-term drug release.

[URETERAL TISSUE ENGINEERING: CHALLENGES AND PROSPECTS].

A broad range of pathologic conditions of the ureter (strictures, obliterations, fistulas, and so on) requiring reconstructive plastic surgery is a challenging urological problem. A variety of approaches to solve the problem indicates the need of searching for new opportunities. A new direction in reconstructive surgery of the ureter is the tissue engineering. Tissue engineering involves the usage of matrices and cells. The matrices can be used both with cultured cells, and without them. This review represents the results of preclinical studies on feasibility of tissue engineering using as a matrix both natural and synthetic materials for different ureter impairments. Presently, there are no data on the use of tissue-engineering for the ureter reconstruction in clinical trials (i.e. involving human subjects). The results of studies presented in the review inspire certain optimism, but ureteral tissue-engineering is a difficult task requiring a balanced approach and well-thought-out design of preclinical studies.

[RISK FACTORS FOR ANTERIOR URETHRAL STRICTURES AFTER TRANSURETHRAL RESECTION OF BENIGN PROSTATIC HYPERPLASIA].

OBJECTIVE: To determine the risk factors for anterior urethral strictures after transurethral resection of benign prostatic hyperplasia. MATERIALS AND METHODS: The present study consists of a prospective and retrospective part. Prospective analysis of 110 patients who had undergone, at the urology clinic of First MSMU named after I.M. Sechenov in the period from January 2009 to February 2014 was performed transurethral resection (TUR) of the prostate. In the retrospective part, the case histories of 85 patients who were treated in the urology clinic First MSMU named after I.M. Sechenov from 2009 to 2013 with a diagnosis of urethral stricture were analysed. Of them, 29 cases urethral stricture occurred earlier after undergoing TURP. Patients from both sides were divided into two groups: group number 1 patients undergoing TURP for BPH who have not formed a urethral stricture, and group number 2 - patients who had urethral stricture formed. RESULTS: According to the criteria for inclusion in group number 1 there were 40 patients who did not form a urethral stricture after TURP and 33 patients were included in the group number 2, in which the late postoperative period was complicated by the formation of a stricture of the urethra. Prostate volume was significantly different in both groups. In group number 1 prostate volume averaged 60 cm3±23 cm3 in group number 2 prostate size equaled an average of 80 cm3±24 cm3. (p<0,05). Having a urethral catheter or cystostomic drainage before surgery was observed in 12% (n=5) in the first group and 27% (n=9) in the second group. Duration of operative benefits was assessed at intervals up to 60 minutes and more than 60 minutes. Thus, the results in the first group are divided into 75% (30) 25% (10), respectively. In the second group, these values were 24,2% (n=8) and 75.8% (n=25). Diabetes mellitus was noted in a first group in 12.5% (n=5) of the patients, in the second group - 30% (n=10). The number of patients seen earlier for hypertension in the group, which did not form a urethral stricture, was 37,5% (n=13) and in the group with urethral strictures - 60,6% (n=20). Having chronic inflammation, which was confirmed upon subsequent morphologic examination of prostate tissue resection, were detected in the first group at 32,5% (n=13) patients and in the group number 2 at 66,6% (n=22) patients and was significantly higher in the second group (p<0,05). CONCLUSIONS: The duration of TURP more than 60 minutes, prostate volume of more than 70 cm3, the presence of diabetes mellitus, and also chronic inflammation of the prostate significantly increases the risk of urethral stricture in the late postoperative period.

[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.

[TISSUE-ENGINEERED SUBSTITUTION URETHROPLASTY BASED ON DECELLULARIZED VASCULAR MATRIX AND AUTOLOGOUS CELLS OF THE BUCCAL MUCOSA: THE FIRST EXPERIENCE].

Urethral strictures and anomalies remain a challenging urological problem. Reconstructive plastic surgery has been shown to be the most effective way to treat them. There are two main types of urethroplasty: anastomosis (anastomotic urethroplasty) and expansion of the urethral lumen using of flaps and grafts (substitution urethroplasty). Currently the ideal material for substitution urethroplasty does not exist. Tissue engineering of the urethra seems to be one of the most promising approaches to address this problem. Various tissues-engineering techniques were proposed for substitution urethroplasty. In this study, tissue-engineering design was based on the decellularized cadaveric arterial wall. The study results demonstrated the feasibility of creating stable tissue-engineered structures with autologous cultured epithelial cells of the buccal mucosa and decellularized matrix from human cadaveric arterial wall (DMCAW). There was a complete engraftment of tissue-engineering design based on DMCAW and buccal mucosa cells, used for substitution urethroplasty in a patient with the bulbar urethral stricture. Postoperatively (within 4 months after surgery) no complications and/or adverse events were observed. However, in the late postoperative period (12 months) there was recurrence of urethral stricture in the middle of the tissue-engineering design and the native urethra that warranted another surgery. Tissue-engineering design based on DMCAW and autologous buccal mucosa is safe as a material for substitution urethroplasty. Further research is required to ascertain the effectiveness of the method.

[Experimental validation of the developing a matrix based on decellularized vascular wall for subsequent substitution urethroplasty].

Urethral strictures are urgent urological problem. Anastomotic and substitution urethroplasty are the most effective treatments. For substitution urethroplasty, buccal mucosa is most often used. There are the following difficulties associated with the substitution urethroplasty: complications in the donor area, the lack of tissue for substitution, an additional incision, and increased timing of surgery due to the need to obtain a flap or graft. Tissue engineering can be useful in solving the above problems. Tissue engineering involves the use a matrix without cells and matrix with one or more types of cells (tissue-engineering designs). In our study we have evaluated the ability to create a matrix for the substitution urethroplasty in animal experiments. The decellularized cadaveric arterial wall was used as a matrix. Decellularization was performed using enzymatic method. At the first stage, we transplanted matrix fragments in interscapular region in rats. An extremely weak bioactivity dof decellularized matrix of cadaveric arterial wall (DMCAW) due to the low immunogenicity of the material was revealed. Thus resorption of DMCAW was quite slow (60-90 days). At the second stage, in an experiment on rabbits, substitution urethroplasty using tubular DMCAW was successfully performed. Intraoperative urethral defect up to 1.8 cm was created, which was replaced by a tubular DMCAW. The use of this type of matrix has showed good structural and functional results: urethral strictures did not arise, the rejection of the matrix was not observed. A slow degradation of the matrix and progressive epithelialization of onnective tissue capsule were revealed. Decellularized matrix based on cadaveric arterial wall can be considered as a material for substitution urethroplasty.

Osteoinducing scaffolds with multi-layered biointerface.

This study was aimed to design and characterise hybrid tissue-engineered constructs composed of osteoinducing polylactide-based scaffolds with multi-layered cellular biointerface for bone tissue reconstruction. Three-dimensional scaffolds with improved hydrophilic and osteoinducing properties were produced using the surface-selective laser sintering (SSLS) method. The designed scaffold pattern had dimensions of 8 × 8 × 2.5 mm and ladder-like pores (∼700 µm in width). Hyaluronic acid-coated polylactide microparticles (∼100 µm in diameter) were used as building blocks and water was used as the photosensitizer for SSLS followed by photocross-linking with Irgacure 2959 photoinitiator. Resulting scaffolds provided successful adhesion and expansion of human bone marrow mesenchymal stromal cells from a single-cell suspension. Induced calcium deposition by the cells associated with osteogenic differentiation was detected in 7-21 days of culturing in basal medium. The values were up to 60% higher on scaffolds produced at a higher prototyping speed under the experimental conditions. Innovative approach to graft the scaffolds with multi-layered cell sheets was proposed aiming to facilitate host tissue-implant integration. The sheets of murine MS-5 stromal cell line exhibited contiguous morphology and high viability in a modelled construct. Thus, the SSLS method proved to be effective in designing osteoinducing scaffolds suitable for the delivery of cell sheets.

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.

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.