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Construction of many tissues and organs de novo requires the use of external epithelial cell sources. In the present study, we optimized the isolation, expansion, and characterization of porcine oral epithelial cells from buccal tissue (Buccal Epithelial Cells, BECs). Additionally, we tested whether key markers [cytokeratin 14 (ck14), p63 protein, and sonic hedgehog molecule (shh)] expression profiles are correlated with three buccal epithelial clone types. Two digestion methods of BECs isolation [Method 1, M1 (collagenase IV/dispase and accutase) and Method 2, M2 (collagenase IV/dispase and trypsin/EDTA)] were compared. Cells obtained by more effective method were further cultured to the third passage and analyzed. Holoclone-, meroclone-, and paraclone-like colonies were identified based on BEC morphology. Immunofluorescent staining was performed to compare selected markers for the indicated buccal clone types. Comparative analysis demonstrated the advantage of isolation using M1 over M2. Cells from the third passage exhibited average 92.73% ± 2.27% presence of ck14. Real-time polymerase chain reaction confirmed expression of tested genes [

Current strategies in urinary bladder augmentation include use of gastrointestinal segments, however, the technique is associated with inevitable complications. An acellular biologic scaffold seems to be a promising option for urinary bladder augmentation. The aim of this study was to evaluate the utility of bladder acellular matrix (BAM) for reconstruction of clinically significant large urinary bladder wall defects in a long-term porcine model. Urinary bladders were harvested from 10 pig donors. Biological scaffolds were prepared by chemically removing all cellular components from urinary bladder tissue. A total of 10 female pigs underwent hemicystectomy and subsequent bladder reconstruction with BAM. The follow-up study was 6 months. Reconstructed bladders were subjected to radiological, macroscopic, histological, immunohistochemical, and molecular evaluations. Six out of ten animals survived the 6-month follow-up period. Four pigs died during observation due to mechanical failure of the scaffold, anastomotic dehiscence between the scaffold and native bladder tissue, or occluded catheter. Tissue engineered bladder function was normal without any signs of postvoid residual urine in the bladder or upper urinary tracts. Macroscopically, graft shrinkage was observed. Urothelium completely covered the luminal surface of the graft. Smooth muscle regeneration was observed mainly in the peripheral graft region and gradually decreased toward the center of the graft. Expression of urothelial, smooth muscle, blood vessel, and nerve markers were lower in the reconstructed bladder wall compared to the native bladder. BAM seems to be a promising biomaterial for reconstruction of large urinary bladder wall defects. Further research on cell-seeded BAM to enhance urinary bladder regeneration is required.

Introduction Human adipose tissue-derived mesenchymal stem/stromal cells (hAT-MSCs) are multipotent stromal cells with a high potential application in tissue engineering and regenerative medicine. Laser irradiation of the place where the cells were implanted can stimulate their proliferation, increase the secretion of growth factors and thus increase the therapeutic effect. Aim To evaluate the influence of two lasers: Er:YAG and diode on the growth of hAT-MSCs in vitro. Material and methods hAT-MSCs were isolated from human subcutaneous adipose tissue. Immunophenotype of hAT-MSCs was confirmed by flow cytometry. Multipotency of hAT-MSCs was confirmed by differentiation into adipogenic, osteogenic and chondrogenic lineages. hAT-MSCs were irradiated with Er:YAG laser (wavelength 2940 nm, frequency 5, 10 Hz, doses: 0.1-1.2 J/cm2) for 2 s and 4 s and diode laser (wavelength 635 nm and doses: 1-8 J/cm2) for 5, 10, 20, 30 and 40 s. Cell viability was analysed 24 h after the exposure using MTT assay. Results Growth stimulation of hAT-MSCs after 5 Hz Er:YAG laser exposure, 0.1 J/cm2 dose for 4 s and 0.3 J/cm2 dose for 4 s was shown in comparison with the control group. Significant growth stimulation of hAT-MSCs after diode laser irradiation in doses of 1-4 J/cm2 was demonstrated compared to the control group. Conclusions The presented results indicate that both lasers, Er:YAG and diode can be used to stimulate stem/stromal cell growth in vitro. The biostimulative effect of laser therapy on stromal cells may be used in the future in aesthetic dermatology in combined laser and cell therapy.

Mumio (Shilajit) is a traditional medicinal drug known and used for hundreds of years. Bladder cancer is one of the most common cancer types and better treatments are needed. This study analysed the in vitro effect of Mumio on urinary bladder cancer cells (T24 and 5637) in comparison to normal uroepithelial cells (SV-HUC1). Cytotoxicity of Mumio was analysed in these cell lines via MTT and real-time cell growth assays as well via the assessment of the cytoskeleton, apoptosis, and cell cycle. Mumio affected the viability of both cell types in a time and concentration dependent manner. We observed a selectivity of Mumio against cancer cells. Cell cycle and apoptosis analysis showed that Mumio inhibited G0/G1 or S phase cell cycle, which in turn induced apoptosis. Our results showed that Mumio was significantly more cytotoxic to urinary bladder cancer cells than to normal cells. These results are promising and indicate Mumio as a great candidate for urinary bladder cancer treatment and further investigations should be performed.

Intestinal segments are currently used in reconstructive urology to create urinary diversion after cystectomy. Ileal conduit (IC) is the dominant type of urinary diversion. Nevertheless, IC is not an ideal solution as the procedure still requires entero-enterostomy to restore the bowel continuity. This step is a source of relevant complications that might prolong recovery time. Fabrication of artificial urinary conduit is a tempting idea to introduce an alternative form of urinary diversion which might improve cystectomy outcomes.The aim of this review is to discuss available research data about artificial urinary conduit and identify major challenges for future studies.Fabrication of artificial urinary conduit is in range of current tissue engineering technology but there are still many challenges to overcome. There is an urgent need for studies to be conducted on large animal models with long follow up to expose the limitation of experimental strategies and to gather data for translational research.

Introduction: Introducing new drugs for clinical application is a very difficult, long, drawn-out, and costly process, which is why drug repositioning is increasingly gaining in importance. The aim of this study was to analyze the cytotoxic properties of ciprofloxacin and levofloxacin on bladder and prostate cell lines in vitro. Methods: Bladder and prostate cancer cell lines together with their non-malignant counterparts were used in this study. In order to evaluate the cytotoxic effect of both drugs on tested cell lines, MTT assay, real-time cell growth analysis, apoptosis detection, cell cycle changes, molecular analysis, and 3D cultures were examined. Results: Both fluoroquinolones exhibited a toxic effect on all of the tested cell lines. In the case of non-malignant cell lines, the cytotoxic effect was weaker, which was especially pronounced in the bladder cell line. A comparison of both fluoroquinolones showed the advantage of ciprofloxacin (lower doses of drug caused a stronger cytotoxic effect). Both fluoroquinolones led to an increase in late apoptotic cells and an inhibition of cell cycle mainly in the S phase. Molecular analysis showed changes in BAX, BCL2, TP53, and CDKN1 expression in tested cell lines following incubation with ciprofloxacin and levofloxacin. The downregulation of topoisomerase II genes (TOP2A and TOP2B) was noticed. Three-dimensional (3D) cell culture analysis confirmed the higher cytotoxic effect of tested fluoroquinolone against cancer cell lines. Conclusions: Our results suggest that both ciprofloxacin and levofloxacin may have great potential, especially in the supportive therapy of bladder cancer treatment. Taking into account the low costs of such therapy, fluoroquinolones seem to be ideal candidates for repositioning into bladder cancer therapeutics.

The use of an ileal segment is a standard method for urinary diversion after radical cystectomy. Unfortunately, utilization of this method can lead to numerous surgical and metabolic complications. This study aimed to assess the tissue-engineered artificial conduit for urinary diversion in a porcine model. Tissue-engineered tubular polypropylene mesh scaffolds were used for the right ureter incontinent urostomy model. Eighteen male pigs were divided into three equal groups: Group 1 (control ureterocutaneostomy), Group 2 (the right ureter-artificial conduit-skin anastomoses), and Group 3 (4 weeks before urostomy reconstruction, the artificial conduit was implanted between abdomen muscles). Follow-up was 6 months. Computed tomography, ultrasound examination, and pyelogram were used to confirm the patency of created diversions. Morphological and histological analyses were used to evaluate the tissue-engineered urinary diversion. All animals survived the experimental procedures and follow-up. The longest average patency was observed in the 3rd Group (15.8 weeks) compared to the 2nd Group (10 weeks) and the 1st Group (5.8 weeks). The implant’s remnants created a retroperitoneal post-inflammation tunnel confirmed by computed tomography and histological evaluation, which constitutes urostomy. The simultaneous urinary diversion using a tissue-engineered scaffold connected directly with the skin is inappropriate for clinical application.

This study developed a new procedure of urinary bladder transplantation on a rat model (n = 40). Heterotopic urinary bladder transplantation (n = 10) in the right groin vessels was performed. Direct urinary bladder examination, microangiography, histological analysis, and India ink injection were performed to evaluate the proposed method's functionality. Observation time was four weeks. One week after the procedure, the graft survival rate was 80%, two urinary bladders were lost due to anastomosis failure. The rest of the grafts survived two weeks without any complications. Lack of transitional epithelium or smooth muscle layer loss and lack of inflammatory process development were observed. This study was performed in order to obtain the necessary knowledge about urinary bladder transplantation. The proposed technique offers a new approach to the existing orthotropic models.

Tissue engineering offers the possibility to overcome limitations of current management for postprostatectomy incontinence and ED. Developed in recent years biotechnological feasibility of mesenchymal stem cell isolation, in vitro cultivation and implantation became the basis for new cell-based therapies oriented to induce regeneration of adult tissue. The perspective to offer patients suffering from post-prostatectomy incontinence or erectile dysfunction minimal invasive one-time procedure utilizing autologous stem cell transplantation is desired management.

Background Evaluating health-related quality of life (HR-QoL) is important in urological oncology. Objectives To evaluate the impact of minimally invasive radical cystectomy (RC) on the patients' QoL depending on the surgical strategy (laparoscopic compared to robotic). Material and methods The study involved 110 patients divided into 2 groups: group 1 (n = 65), qualified for robotic-assisted RC (RARC), and group 2 (n = 45), qualified for laparoscopic RC (LRC). A prospective analysis of QoL was performed. In the study, we used sociodemographic data taken from the patients' medical records, and data from standardized questionnaires of QoL surveys entitled Functional Assessment of Cancer Therapy - General (FACT-G) and Functional Assessment of Cancer Therapy for Bladder Cancer (FACT-Bl). Results In the patients who had undergone LRC, a statistically significant difference in HR-QoL was noted only for the subjective well-being (SWB) domain regarding family and social life: The patients had higher SWB values before surgery than after it. In the patients who had undergone RARC, statistically significant differences in HR-QoL were noted in 3 domains: 1) SWB - family and social life, 2) FACT-Bl assessment and 3) FACT-G assessment. Both before and after surgery, no statistically significant differences were found between the 2 groups for any of the HR-QoL domains. Conclusions The surgical technique of minimally invasive endoscopic RC used (laparoscopic or robotic) does not affect HR-QoL domains.

Tissue engineering allows to combine biomaterials and seeded cells to experimentally replace urinary bladder wall. The normal bladder wall however, includes branched neuronal network propagating signals which regulate urine storage and voiding. In this study we introduced a novel biocomposite built from amniotic membrane (Am) and graphene which created interface between cells and external stimuli replacing neuronal network. Graphene layers were transferred without modifying Am surface. Applied method allowed to preserve the unique bioactive characteristic of Am. Tissue engineered constructs composed from biocomposite seeded with smooth muscle cells (SMC) derived from porcine detrusor and porcine urothelial cells (UC) were used to evaluate properties of developed biomaterial. The presence of graphene layer significantly increased electrical conductivity of biocomposite. UCs and SMCs showed an organized growth pattern on graphene covered surfaces. Electrical filed stimulation (EFS) applied in vitro led additionally to increased SMCs growth and linear arrangement. 3D printed chamber equipped with 3D printed graphene based electrodes was fabricated to deliver EFS and record pressure changes caused by contracting SMCs seeded biocomposite. Observed contractile response indicated on effective SMCs stimulation mediated by graphene layer which constituted efficient cell to biomaterial interface.

Long-term culture of mesenchymal stromal/stem cells in vitro leads to their senescence. It is very important to define the maximal passage to which the mesenchymal stromal/stem cells maintain their regenerative properties and can be used for cellular therapies and construction of neo-organs for clinical application. Adipose-derived stromal/stem cells were isolated from porcine adipose tissue. Immunophenotype, population doubling time, viability using bromodeoxyuridine assay, MTT assay, clonogencity, β-galactosidase activity, specific senescence-associated gene expression, apoptosis, and cell cycle of adipose-derived mesenchymal stromal/stem cells (AD-MSCs) were analyzed. All analyses were performed through 12 passages (P). Decreasing viability and proliferative potential of AD-MSCs with subsequent passages together with prolonged population doubling time were observed. Expression of β-galactosidase gradually increased after P6. Differentiation potential of AD-MSCs into adipogenic, chondrogenic, and osteogenic lineages decreased at the end of culture (P10). No changes in the cell cycle, the number of apoptotic cells and expression of specific AD-MSC markers during the long-term culture were revealed. Molecular analysis showed increased expression of genes involved in activation of inflammatory response. AD-MSCs can be cultured for in vivo applications without loss of their properties up to P6.

Purpose Minimally invasive techniques have had an increasing implementation trend in recent years but they are associated with significantly higher costs. Objective To assess the costs of radical cystectomy procedure performed using a laparoscopic (LRC) and robot-assisted (RARC) technique and to determine the postoperative complications resulting from surgical treatment and estimate the impact of complications on the formation of total costs. Materials and Methods A retrospective costs analysis was conducted between February 2016 and March 2018 in 89 patients who underwent radical cystectomy with ureterocutaneostomy or ileal urinary diversion. Primary outcomes included patients’ characteristics and variables which had an influence on the overall cost of the surgery. Secondary outcomes included selected direct medical costs: hospitalization, operating room, blood products, parenteral nutrition, diagnostic procedures and costs of surgical instruments. Results Perioperative data were similar in the compared groups. The Clavien-Dindo complications rate ≤30 days was lower for RARC. LRC had a higher mean length of hospital stay than RARC but a shorter operating time by an average of 66 min. The shorter operating time had an impact on the operating room costs (LRC: €769; RARC: €1015; p Conclusions In the Polish healthcare system RARC had a higher total cost but a shorter length of stay, a lower volume of blood transfusions and a lower percentage complications rates than LRC. The cost analysis presented in this paper showed the superiority of LRC over RARC.

The aim of this paper was to collect currently available data related to the use of stem cells in aesthetic dermatology and plastic surgery based on a systemic review of experimental and clinical applications. We found that the use of stem cells is very promising but the current state of art is still not effective. This situation is connected with not fully known mechanisms of cell interactions, possible risks and side effects. We think that there is a big need to create and conduct different studies which could resolve problems of stem cells use for implementation into aesthetic dermatology and plastic surgery.

Introduction: Urinary tract regeneration using tissue engineering is one of the most challenging issues in the field of reconstructive urology. Cells seeded on scaffold are exposed to urine immediately after the implantation. The outcome of urinary bladder regeneration is depended on the ability of these cells to survive, proliferate and regenerate. The aim of this study was to compare a sensitivity of three different cell lines to urine in vitro. Material and methods: Three different cell lines were isolated from porcine bladder (urothelial cells, UCs and smooth muscle cells, SMCs) and adipose tissue (adipose-derived stem cells, ADSCs). Cell viability (MTT assay), proliferation (real-time cell analysis using xCELLigence system) and apoptosis/necrosis (flow cytometry) were analyzed after exposition to urine. Results: ADSCs were the most sensitive to urine compared to two other tested cell lines. Among the bladder cell lines the urothelial cells were more resistant to urine than SMCs. Twenty four hour incubation of UCs, SMCs and ADSCs with urine lead to ∼40%, ∼70% and ∼90% reduction of their viability, respectively. The mechanism of urine mediated cytotoxicity differed depending on the tested cell type. Conclusion: Urothelial and smooth muscle cells seem to be more suitable for urinary bladder regeneration compared to mesenchymal stem cells, however these cells have limited application especially in the case of urinary bladder cancer.

Regenerative medicine is a new branch of medicine based on tissue engineering technology. This rapidly developing field of science offers revolutionary treatment strategy aimed at urinary bladder regeneration. Despite many promising announcements of experimental urinary bladder reconstruction, there has been a lack in commercialization of therapies based on current investigations. This is due to numerous obstacles that are slowly being identified and precisely overcome. The goal of this review is to present the current status of research on urinary bladder regeneration and highlight further challenges that need to be gradually addressed. We put an emphasis on expectations of urologists that are awaiting tissue engineering based solutions in clinical practice. This review also presents a detailed characteristic of obstacles on the road to successful urinary bladder regeneration from urological clinician perspective. A defined interdisciplinary approach might help to accelerate planning transitional research tissue engineering focused on urinary tracts.

Tissue engineering requires suitable polymeric scaffolds, which act as a physical support for regenerated tissue. A promising candidate might be polyurethane (PUR) scaffold, due to the ease of the PUR properties design, which can be adjusted directly to the intended purpose. In this study, we report a successful fabrication of porous polyurethane scaffolds (PPS) using solvent casting/particulate leaching technique combined with thermally induced phase separation. The obtained PPS had comparable chemical structure to native PUR, which was confirmed by FTIR and HNMR analyses. The performed DSC study determined a similar T g of the obtained PPS to native PUR (−38 °C). The analysis of TEM micrographs revealed that PPS had a homogenous structure. The studied PPS interactions with canola oil, distilled water, saline solution and phosphate-buffered saline after 3 months of incubation revealed that these materials have stable character in these media. The significant decrease of contact angle from 68° for native PUR to 54° for PPS was noted, as well as the decrease of mechanical properties (T Sb ~ 1 MPa and e b ~ 95% of PPS were comparable to the native aorta tissue of T Sb ~ 0.3–0.8 MPa and e b ~ 50–100%). Through SEM analysis, the morphology of the PPS was determined: the porosity was 87% and the pore sizes in the range of 98–492 µm. The biological studies revealed that the obtained PPS are sensitive to microorganisms such as Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli and that they are biocompatible with the 3T3 NIH cell line. In summary, the obtained PPS scaffolds may be a suitable material for soft tissue engineering like blood vessels.

BACKGROUND Electrospun nanofibers have widespread putative applications in the field of regenerative medicine and tissue engineering. When compared to naturally occurring collagen matrices, electrospun nanofiber scaffolds have two distinct advantages: they do not induce a foreign body reaction and they are not at risk for biological contamination. However, the exact substrate, structure, and production methods have yet to be defined. MATERIAL AND METHODS In the current study, tubular-shaped poly(L-lactide-co-caprolactone) (PLCL) constructs produced using electrospinning technology were evaluated for their potential application in the field of tissue regeneration in two separate anatomic locations: the skin and the abdomen. The constructs were designed to have an internal diameter of 3 mm and thickness of 200 μm. Using a rodent model, 20 PLCL tubular constructs were surgically implanted in the abdominal cavity and subcutaneously. The constructs were then evaluated histologically using electron microscopy at 6 weeks post-implantation. RESULTS Histological evaluation and analysis using scanning electron microscopy showed that pure scaffolds by themselves were able to induce angiogenesis after implantation in the rat model. Vascularization was observed in both tested groups; however, better results were obtained after intraperitoneal implantation. Formation of more and larger vessels that migrated inside the scaffold was observed after implantation into the peritoneum. In this group no evidence of inflammation and better integration of scaffold with host tissue were noticed. Subcutaneous implantation resulted in more fibrotic reaction, and differences in cell morphology were also observed between the two tested groups. CONCLUSIONS This study provides a standardized evaluation of a PLCL conduit structure in two different anatomic locations, demonstrating the excellent ability of the structure to achieve vascularization. Functional, histological, and mechanical data clearly indicate prospective clinical utilization of PLCL in critical size defect regeneration.

Background Molecular mechanisms underlying the regenerative process induced by stem cells in tissue-engineered urinary bladder are poorly explained. The study was performed to explore the pathways associated with regeneration process in the urinary bladder reconstructed with adipose tissue-derived mesenchymal stromal cells (ASCs). Methods Rat urinary bladders were reconstructed with bladder acellular matrix (BAM) (n = 52) or BAM seeded with adipose tissue-derived mesenchymal stromal cells (ASCs) (n = 52). The process of bladder healing was analyzed at 7, 30, 90, and 180 days postoperatively using macroscopic histologic and molecular techniques. Gene expression was analyzed by microarrays and confirmed by real-time PCR. Results Numerous differentially expressed genes (DEGs) were identified between the bladders augmented with BAM seeded with ASCs or BAM only. Pathway analysis of DEGs allows to discover numerous pathways among them Hedgehog, TGF-β, Jak-STAT, PI3-Akt, and Hippo modulated by ASCs during the healing process of tissue-engineered urinary bladder. Real-time PCR analysis confirmed upregulation of genes involved in the Hedgehog signaling pathway including Shh, Gli1, Smo, Bmp2, Bmp4, Wnt2, Wnt2b, Wnt4, Wnt5a, and Wnt10 in urinary bladders reconstructed with ASC-seeded grafts. Conclusion The study provided the unequivocal evidence that ASCs change the molecular pattern of healing in tissue-engineered urinary bladder and indicated which signaling pathways triggered by ASCs can be associated with the regenerative process. These pathways can be used as targets in the future studies on induced urinary bladder regeneration. Of particular interest is the Hedgehog signaling pathway that has been upregulated by ASCs during healing of tissue-engineered urinary bladder. Electronic supplementary material The online version of this article (10.1186/s13287-019-1266-1) contains supplementary material, which is available to authorized users.

The causes of bladder cancer are not yet fully uncovered, however the research has identified a number of factors that may increase the risk of developing this cancer. The chemical carcinogenesis of bladder cancer due to chronic exposure to aromatic hydrocarbons has been well-established. The identification of this correlation led to an improvement of safety measures in chemical industry and a gradual decrease of bladder cancer cases among workers. Nevertheless, in the majority of bladder cancer cases, the specific cause of the disease still can't be specified. It makes the question of unrecognized factors associated with bladder cancer development even more relevant. Taking under consideration known chemical carcinogenesis of bladder cancer, this minireview takes under investigation the possible link between using scented candles and a risk of bladder cancer development. Burning scented candles contain many of the substances that are associated with a bladder cancer. Furthermore the scented candles are not only very popular but also widely available on the market, with limited quality regulations and unspecified raw materials determining a spectrum of potentially dangerous substances emitted during burning.

Pancreatic islet β cells synthesize the hormone insulin, which is fundamental for the control of glucose metabolism. β cells can be lost as the result of an autoimmune process, leading to type 1 diabetes (T1D), they can develop functional impairments in the context of insulin resistance (type 2 diabetes), or they can be impaired owing to genetic conditions compromising their development or function (monogenic diabetes). Cell-based therapies for beta-cell replacement are under investigation. Transplantation of cadaveric pancreas or pancreatic islets can correct diabetes, but the low numbers of organs available and the need for immunosuppression limit these strategies. β cells can be derived from embryonic stem cells, induced pluripotent stem cells, or adult stem/progenitor cells. Novel findings indicate that stem/progenitor cells exist in the adult human pancreas. Differentiation strategies are largely based on knowledge from embryology. Other regenerative strategies include reprogramming or the delivery of prodifferentiation signals. This chapter will also cover studies of mesenchymal stem cell (MSC) transplantation in diabetic patients. MSCs can modulate immunity and preserve or even stimulate the regeneration of β cells. MSCs had a good safety profile in initial clinical trials for T1D and are being tested as a therapeutic option for complications of diabetes mellitus.

Tissue engineering is focusing research effort on search for new biomaterials that might be applied to create artificial urinary conduit. Nevertheless, the demanding biomechanical characteristics necessary for proper conduit function is difficult to be replicated. In this study, we are introducing novel marine biomaterial obtained by decellularization of squid mantle derived from Loligo vulgaris. Squid mantles underwent decellularization according to developed dynamic flow two-staged procedure. Efficacy of the method was confirmed by computational dynamic flow analysis. Subsequently Decellularized Squid Mantle (DSM) underwent extensive histological analysis and mechanical evaluation. Based on gained biomechanical data the computational modelling using finite element method was utilized to simulate behavior of DSM used as a urinary conduit. Taking into account potential application in reconstructive urology, the DSM was then evaluated as a scaffold for urothelial and smooth muscle cells derived from porcine urinary bladder. Conducted analysis showed that DSM created favorable environment for cells growth. In addition, due to polarized structure and natural external polysaccharide layer, it protected seeded cells from urine.

Clinical pancreatic islet transplantation can be considered one of the safest and least invasive transplant procedures. Remarkable progress has occurred in both the technical aspects of islet cell processing and the outcomes of clinical islet transplantation. With1,500 patients treated since 2000, this therapeutic strategy has moved from a curiosity to a realistic treatment option for selected patients with type 1 diabetes mellitus (that is, those with hypoglycaemia unawareness, severe hypoglycaemic episodes and glycaemic lability). This Review outlines the techniques required for human islet isolation, in vitro culture before the transplant and clinical islet transplantation, and discusses indications, optimization of recipient immunosuppression and management of adjunctive immunomodulatory and anti-inflammatory strategies. The potential risks, long-term outcomes and advances in treatment after the transplant are also discussed to further move this treatment towards becoming a more widely available option for patients with type 1 diabetes mellitus and eventually a potential cure.

Many experimental approaches have been conducted in order to isolate urothelial cells from bladder tissue biopsies, but each method described has utilized different protocols and sources of bladder tissue. In this study, we compared the different methods of urothelial cell isolation available in literature together with standardized methods in order to obtain more unified results. Five methods for primary porcine urothelial culture establishment were compared: tissue explants and four enzymatic methods utilizing collagenase II, dispase II, combination of dispase II and trypsin, and trypsin alone. The average number of isolated cells, cell morphology, success of established culture, average number of cells from the first passage, expression of p63 and pancytokeratin and the characterization of urothelial cell growth, and aging were analyzed during the in vitro culture. The method utilizing dispase II was the most efficient and reproducible method for the isolation and culture of porcine urothelial cells when compared to the other tested methods. Urothelial cells obtained by this method grew considerably well and the cultures were established with high efficiency, which enabled us in obtaining a large quantity of cells with normal morphology. Contamination with fibroblasts in this method was the lowest. The utilization of a proper method for urothelial cell isolation is a critical step in the urinary tract regeneration when using tissue engineering techniques. In summary, this study demonstrated that by utilizing the described method with dispase II, a suitable number of cells was achieved, proving the method useful for tissue regeneration.

On the 60th anniversary of the first successfully performed renal transplantation, we summarize the historical, current and potential future status of kidney transplantation. We discuss three different aspects with a potential significant influence on kidney transplantation progress: the development of surgical techniques, the influence of regenerative medicine and tissue engineering, and changes in immunosuppression. We evaluate the standard open surgical procedures with modern techniques and compare them to less invasive videoscopic as well as robotic techniques. The role of tissue engineering and regenerative medicine as a potential method for future kidney regeneration or replacement and the interesting search for novel solutions in the field of immunosuppression will be discussed. After 60 years since the first successfully performed kidney transplantation, we can conclude that the greatest achievements are associated with the development of surgical techniques and with planned systemic immunosuppression.

The use of FDM/FFF in 3D printing for medical sciences is becoming common. This is due to the high availability and decent price of both 3D printers and filaments useful for FDM/FFF. Currently, researchers' attention is focused mainly on the study of medical filaments based on PLA, PCL or their modifications. This contributes to insufficient diversity of medical-grade filaments on the market. Moreover, due to the lack of specified standards for filaments testing, manufacturers often provide merely the characteristics of the raw materials, which were used for filaments fabrication. This lack of comprehensive data can be problematic when viewed as medical-grade material. As a consequence of this overview, we have performed a comprehensive evaluation of a flexible medical-grade filament for FDM/FFF 3DP - Bioflex® (Filoalfa). We have performed complex characterization through a variety of methods and techniques including spectroscopic analysis (FTIR, Raman), dynamic mechanical analysis (DMA), thermal properties (DSC, TGA), rheological characteristic (MFR). In the next step, printed Bioflex® samples were utilized to characterize the material behaviour after the 3D printing process. The mechanical analysis allowed to estimate how the material strength decreases after the printing process according to the values given in the technical data sheet. The contact angle measurements determined wettability of the Bioflex® printouts. Performed series of in vitro studies were carried out to assess its potential as as implantable structures. In conclusion, 3D printing process did not affect the printouts biocompatibility (ISO 10993:5). Accelerated degradation studies indicated elevated hydrolysis resistance of printed samples. In turn, performed incubation in simulated body fluid (SBF) solution, revealed carbonated hydroxyapatite (HAp) deposition on printouts surface indicating their bioactive properties. Thus, studied filament seems to be a suitable candidate for further development of FDM/FFF 3DP structures for advanced biological and medical application.

The chemical reaction between cis-[PtCl2(dmso)2] and kinetin riboside (KR) or 1-β- d -ribofuranosyl-4-(2-pyridyl)-1H-1,2,3-triazole (PTR) has resulted in two novel platinum(II) complexes being obtained with different compositions of the coordination sphere: cis-[PtCl2(PTR)] (1) and cis-[PtCl2(dmso)(KR)] (2). Based on multinuclear NMR results (1H, 13C, 15N, 195Pt), we have demonstrated that the N-donor ligands PTR and KR are able to coordinate to the Pt(II) ion as bidentate via two nitrogen atoms (N(3) and N(7)) or monodentate via only one nitrogen atom (N(7)), respectively. On the other hand, biological studies showed that the novel platinum(II) complexes exhibit different in vitro cytotoxicity towards A549 (lung carcinoma epithelial cells), T24 (urinary bladder cancer cells) and CRL1872 (malignant melanoma cells). The complex cis-[PtCl2(dmso)(KR)] (2) shows in vitro cytotoxicity against T24 and CRL1872 cell lines (the IC50 parameters amount to 52.2 μM and 21.4 μM, respectively), whereas cis-[PtCl2(PTR)] (1) is not able to inhibit a proliferation of these cells in the range of tested concentrations.

Introduction: In reconstructive urology, autologous tissues such as intestinal segments, skin, and oral mucosa are used. Due to their limitations, reconstructive urologists are waiting for a novel material, which would be suitable for urinary tract wall replacement. Human amniotic membrane (AM) is a naturally derived biomaterial with a capacity to support reepithelization and inhibit scar formation. AM has a potential to become a considerable asset for reconstructive urology, i.e., reconstruction of ureters, urinary bladder, and urethrae. Areas covered: This review aims to discuss the potential application of human AM in reconstructive urology. The environment for urinary tract healing is particularly unfavorable due to the presence of urine. Due to its fetal origin, the bioactivity of AM is orientated to induce intrinsic regeneration mechanisms and inhibit scarring. This review introduces the concept of applying human AM in reconstructive urology procedures to improve their outcomes and future tissue engineering based strategies. Expert opinion: Many fields of medicine that have accomplished translational research have proven the usefulness of AM in clinical practice. There is an urgent need for studies to be conducted on large animal models that might convincingly demonstrate the underestimated potential of AM to urologists around the world.

Background The tissue engineering of urinary bladder advances rapidly reflecting clinical need for a new kind of therapeutic solution for patients requiring urinary bladder replacement. Majority of the bladder augmentation studies have been performed in small rodent or rabbit models. Insufficient number of studies examining regenerative capacity of tissue-engineered graft in urinary bladder augmentation in a large animal model does not allow for successful translation of this technology to the clinical setting. The aim of this study was to evaluate the role of adipose-derived stem cells (ADSCs) in regeneration of clinically significant urinary bladder wall defect in a large animal model. Methods ADSCs isolated from a superficial abdominal Camper’s fascia were labeled with PKH-26 tracking dye and subsequently seeded into bladder acellular matrix (BAM) grafts. Pigs underwent hemicystectomy followed by augmentation cystoplasty with BAM only (n = 10) or BAM seeded with autologous ADSCs (n = 10). Reconstructed bladders were subjected to macroscopic, histological, immunofluoresence, molecular, and radiological evaluations at 3 months post-augmentation. Results Sixteen animals (n = 8 for each group) survived the 3-month follow-up without serious complications. Tissue-engineered bladder function was normal without any signs of post-voiding urine residual in bladders and in the upper urinary tracts. ADSCs enhanced regeneration of tissue-engineered urinary bladder but the process was incomplete in the central graft region. Only a small percentage of implanted ADSCs survived and differentiated into smooth muscle and endothelial cells. Conclusions The data demonstrate that ADSCs support regeneration of large defects of the urinary bladder wall but the process is incomplete in the central graft region. Stem cells enhance urinary bladder regeneration indirectly through paracrine effect. Electronic supplementary material The online version of this article (10.1186/s13287-018-1070-3) contains supplementary material, which is available to authorized users.

Introduction Prostate cancer (PCa) is the most common type of cancer among men in Europe. Current recommendations for screening and diagnosis are based on prostate-specific antigen (PSA) measurements and the digital rectal examination (DRE). Both of them are triggers for prostate biopsy. Limited specificity of the PSA test brings, however, a need to develop new, better diagnostic tools. Several commercially available variations of the PSA test including: prostate health index (PHI), 4Kscore as well as molecular PCA3 score, have already revealed its value, lowering the number of unnecessary biopsies. Material and methods This review summarizes published results of the current most promising, clinically proven and experimentally evaluated PCa biomarkers which have potential for creation of new diagnostic tests. Results In the last few years new approaches for providing significantly better biomarkers, an alternative to PSA, have been introduced. Modern biomarkers show improvement in being used as not only a diagnosis procedure, but also for staging, evaluating aggressiveness and managing the therapeutic process. The most promising group are molecular markers, among them microRNAs(miRNAs) and long noncoding RNAs (lncRNAs) are most frequent. Their superiority, over standard PSA, in predicting tumor formation in early stages, and clinically non-symptomatic metastases has been noticed. Extracellular vesicles presence in biofluids have brought focus of many research groups, indicating their potential significance. This group of nanoparticles has potential not only in diagnostic and therapy management process, but also as a potential therapeutic target. Conclusions Finding better PCa biomarkers, replacing the current PSA measurement, is firmly needed in modern urology practice.

Urothelial cell populations which differ in morphology and proliferation capacities can be isolated from the urinary bladder. The goal of this study was to analyze a clonal, proliferative, and self-renewing potential of porcine urothelial cells and to compare expression of selected adhesion and tight junction molecules, urothelial and stem cell markers for the urothelial clone types. Urothelial cells were isolated from 10 porcine urinary bladders. Three different clone types: holoclone-, meroclone-and paraclone-like colonies were identified based on their morphology. To characterize and compare the urothelial clones the immunofluorescent stains were performed. Expression of pancytokeratin (PanCK), Ki-67 and p63 was higher for holoclone- like cells compared to meroclone-and paraclone-like cells (P 0.05). Meroclone-like cells expressed higher levels of p63 compared to paraclone- like cells (P 0.05). The level of Ki-67 and PanCK for meroclone- and paraclone- like cells was comparable (P 0.05). β1 and β4 integrins were not expressed. Expression of zonula occludens-1 (ZO-1) in cell-cell junctions for paraclone-, meroclone-and holoclone-like cells was 17.6 ± 0.6, 14.7 ± 0.5, and 16.1 ± 0.4, respectively. The results of actin filaments (F-actin) expression were 253,634 ± 6,920 for meroclone-like cells, 198,512 ± 7,977 for paraclone-like cells and 133,544 ± 3,169 for holoclone-like cells. Three urothelial cell types with differing features can be isolated from the bladder. Holoclone-like cells are the richest in stem cells and should be used in further studies for construction of neo-bladder or neo-conduit using tissue engineering methods.

In this paper we created novel poly(ester urethane)s (PESUs) designed specifically for tissue engineering. The PESUs were derived from oligomeric α,ω-dihydroxy(ethylene-butylene adipate) (dHEBA), 1,4-butanediol (BDO) and aliphatic 1,6-hexamethylene diisocyanate (HDI) and modified with l-ascorbic acid to improve their biocompatibility. In addition, we determined their mechanical properties (such as tensile strength, elongation at break and hardness). To determine how these materials may potentially behave after implementation in tissue, we estimated the degradation behavior of obtained PESUs in various chemical environments, which were exemplified by canola oil, saline solution, distilled water and phosphate buffered saline (PBS). Finally, the PESU biocompatibility and hemocompatibility tests were carried out. Biocompatibility was determined with a MTT assay and it was performed with 3T3 cell line. The results showed satisfactory mechanical properties, good hemocompatibility and improved biocompatibility, which let us to conclude that PESUs modified with ascorbic acid might find an application in biomedical field of soft tissues engineering.

Pancreatic islet implantation has been recently shown to be an efficient method of treatment for type 1 diabetes. However, limited availability of donor islets reduces its use. Bone morrow would provide potentially unlimited source of stem cells for generation of insulin-producing cells. This study was performed to evaluate the influence of extracellular matrix proteins like collagen, laminin, and vitronectin on bone marrow mesenchymal stem cells (BM-MSCs) transdifferentiation into islet-like cells (ILCs) in vitro. To our knowledge, this is the first report evaluating the importance of vitronectin in transdifferentiation of BM-MSCs into ILCs. Rat BM-MSCs were induced to ILCs using four-step protocol on plates coated with collagen type IV, laminin type I and vitronectin type I. Quantitative real-time PCR was performed to detect gene expression related to pancreatic β cell development. The induced cells expressed islet-related genes including: neurogenin 3, neurogenic differentiation 1, paired box 4, NK homeobox factor 6.1, glucagon, insulin 1 and insulin 2. Laminin but not collagen type IV or vitronectin enhanced expression of insulin and promoted formation of islet-like structures in monolayer culture. Laminin triggered transdifferentiation of BM-MSCs into ILCs.

In recent years, urine has emerged as a source of urine cells. Two different types of cells can be isolated from urine: urine derived stem cells (USCs) and renal tubular cells called urine cells (UCs). USCs have great differentiation properties and can be potentially used in genitourinary tract regeneration. Within this paper, we attempt to demonstrate that such as easily accessible source of cells, collected during completely non-invasive procedures, can be better utilized. Cells derived from urine can be isolated, stored, and used for the creation of urine stem cell banks. In the future, urine holds great potential to become a main source of cells for tissue engineering and regenerative medicine.

The drug-carrier system used as innovative haemostatic dressing with oncostatic action is studied. It is obtained from CDDP (cisplatin) doped SWCNT (single walled carbon nanotubes), modified and purified by H2O2 in hydrothermal treatment process. In the in vivo nephron sparing surgery (NSS) study we used 35 BALB/c nude mice with induced renal cancer using adenocarcinoma 786-o cells. Animals were divided into four groups: CDDP(M-), CDDP(M+), CONTROL(M-) and CONTROL(M+). In CDDP(M-) and CDDP(M+) groups we used, intraoperatively, carbon nanotubes filled with cisplatin (CDDP). In CONTROL(M-) and CONTROL(M+) groups carbon nanotubes were used alone. During NSS free margin (M-) or positive margin (M+) was performed. In the CDDP(M-) group, we do not observe local tumor recurrences. In Group CDDP(M+) only one animal was diagnosed with tumor recurrence. In control groups the recurrent tumor formation was observed. In our study, it is shown that CDDP filled SWCNT inhibit cancer recurrence in animal model NSS study, and can be successfully applied as haemostatic dressings for local chemoprevention.

// Maciej Nowacki 1 , Margarita Peterson 2 , Tomasz Kloskowski 3 , Eleanor McCabe 2 , Delia Cortes Guiral 4 , Karol Polom 5, 6 , Katarzyna Pietkun 7 , Barbara Zegarska 7 , Marta Pokrywczynska 3 , Tomasz Drewa 3 , Franco Roviello 7 , Edward A. Medina 8 , Samy L. Habib 9, 10 and Wojciech Zegarski 1 1 Chair of Department of Surgical Oncology, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Oncology Centre of Franciszek Łukaszczyk Memorial Hospital, Bydgoszcz, Poland 2 Department of Plastic and Reconstructive Surgery, Wake Forest School of Medicine, Winston-Salem, NC, USA 3 Chair of Urology, Department of Regenerative Medicine, Ludwik Rydygier’s Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Torun, Poland 4 Department of General Surgery (Peritoneal Surface Surgical Oncology), Fundacion Jimenez Diaz Hospital, Madrid, Spain 5 General Surgery and Surgical Oncology Department, University of Siena, Siena, Italy 6 Department of Surgical Oncology, Medical University of Gdansk, Gdansk, Poland 7 Chair of Cosmetology and Aesthetic Dermatology, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun. Bydgoszcz, Poland 8 Department of Pathology, University of Texas Health, San Antonio, TX, USA 9 Department of Cell Systems and Anatomy, University of Texas Health Geriatric Research Education, San Antonio, TX, USA 10 South Texas Veterans Health Care System, San Antonio, TX, USA Correspondence to: Maciej Nowacki, email: maciej.s.nowacki@gmail.com Samy L. Habib, email: habib@uthscsa.edu Keywords: CRS + HIPEC, HIPEC, PIPAC, peritoneal carcinomatosis, nanomedicine Received: April 21, 2017 Accepted: August 15, 2017 Published: August 31, 2017 ABSTRACT The treatment of peritoneal surface malignances has changed considerably over the last thirty years. Unfortunately, the palliative is the only current treatment for peritoneal carcinomatosis (PC). Two primary intraperitoneal chemotherapeutic methods are used. The first is combination of cytoreductive surgery (CRS) and Hyperthermic IntraPEritoneal Chemotherapy (HIPEC), which has become the gold standard for many cases of PC. The second is Pressurized IntraPeritoneal Aerosol Chemotheprapy (PIPAC), which is promising direction to minimally invasive as safedrug delivery. These methods were improved through multicenter studies and clinical trials that yield important insights and solutions. Major method development has been made through nanomedicine, specifically nanoparticles. Here, we are presenting the latest advances of nanoparticles and their application to precision diagnostics and improved treatment strategies for PC. These advances will likely develop both HIPEC and PIPAC methods that used for in vitro and in vivo studies. Several benefits of using nanoparticles will be discussed including: 1) Nanoparticles as drug delivery systems; 2) Nanoparticles and Near Infrred (NIR) Irradiation; 3) use of nanoparticles in perioperative diagnostic and individualized treatment planning; 4) use of nanoparticles as anticancer dressing’s, hydrogels and as active beeds for optimal reccurence prevention; and 5) finally the curent in vitro and in vivo studies and clinical trials of nanoparticles. The current review highlighted use of nanoparticles as novel tools in improving drug delivery to be effective for treatment patients with peritoneal carcinomatosis.

Background: Research is scarce regarding the effectiveness of dermal fillers containing autologous stem cells. Objectives: The authors sought to determine the local and systemic effects of adipose-derived stem cells (ADSCs) as a component of dermal fillers in an animal model. Methods: Wistar rats were injected with 1 of the following dermal fillers: ADSCs combined with hyaluronic acid (ADSC-HA), ADSCs combined with fish collagen (ADSC-COL), HA alone (CONTROL-HA), or COL alone (CONTROL-COL). Fillers were injected into the glabella, dorsum, and chest of each animal. The ADSCs were labeled with PKH26 to assess cell migration. Filling effects (FEs) were measured immediately after injection and at 1.5 months and 3 months after injection. Skin specimens were stained with hematoxylin and eosin to assess localization and persistence of ADSCs. Results: Mean FEs in animals implanted with ADSCs were greater and persisted longer than those of controls. No inflammatory responses were observed in any group. Three months after injection, PKH26-positive cells comprised nearly 70% of cells at the injection site in animals treated with ADSC-HA. PKH26 fluorescence also was detected in the spleen but not in the brain, kidney, or lung. Conclusions: Stem cells have the potential to improve the aesthetic effects and longevity of dermal fillers.

OBJECTIVES This study was designed to evaluate the impact of fructose-rich diet and chronic kidney disease (CKD) on the in vitro function of pancreatic islets. METHODS Fifty-four rats were divided into 3 equal groups as follows: control, rats with CKD 1/2 that underwent surgical uninephrectomy, and rats with CKD 5/6 that underwent uninephrectomy and kidney cortex mass resection. Each group was further assigned to 3 diet protocols--regular diet, regular diet with 10% fructose (F10), and 60% fructose-rich diet (F60). After 8 weeks of insulin administration, C-peptide, glycated hemoglobin level, serum urea nitrogen, creatinine clearance, and homeostasis model assessment of insulin resistance were evaluated. Static glucose insulin stimulation test of isolated pancreatic islets and histologic analysis of pancreatic tissue were performed. RESULTS The F10 diet increased the levels of insulin and C-peptide in all groups. Homeostasis model assessment of insulin resistance was increased in all animals fed with fructose. The elevated levels of creatinine and diminished creatinine clearance were detected in CKD 5/6 rats fed with 60% fructose-rich diet. The F10 diet resulted in high levels of serum insulin and C-peptide and glucose-stimulated insulin secretion. Fructose-rich diet increased the islet size and number, with irregular morphology and exocrine tissue fibrosis. CONCLUSIONS The fructose-rich diet accelerates the progression of CKD and affects the pancreatic islet function.