Mesenchymal stromal cells modulate the molecular pattern of healing process in tissue-engineered urinary bladder: the microarray data.

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-ß, 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.

Understanding the role of mesenchymal stem cells in urinary bladder regeneration-a preclinical study on a porcine model.

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.

Are agricultural and natural sources of bio-products important for modern regenerative medicine? A review.

[b] Abstract Introduction and objectives[/b]. As tissue engineering and regenerative medicine have continued to evolve within the field of biomedicine, the fundamental importance of bio-products has become increasingly apparent. This true not only in cases where they are derived directly from the natural environment, but also when animals and plants are specially bred and cultivated for their production. [b]Objective.[/b] The study aims to present and assess the global influence and importance of selected bio-products in current regenerative medicine via a broad review of the existing literature. In particular, attention is paid to the matrices, substances and grafts created from plants and animals which could potentially be used in experimental and clinical regeneration, or in reconstructive procedures. [b]Summary.[/b] Evolving trends in agriculture are likely to play a key role in the future development of a number of systemic and local medical procedures within tissue engineering and regenerative medicine. This is in addition to the use of bio-products derived from the natural environment which are found to deliver positive results in the treatment of prospective patients.

Does the Mesenchymal Stem Cell Source Influence Smooth Muscle Regeneration in Tissue-Engineered Urinary Bladders?

A variety of tissue engineering techniques utilizing different cells and biomaterials are currently being explored to construct urinary bladder walls de novo, but so far no approach is clearly superior. The aim of this study was to determine whether mesenchymal stem cells (MSCs) isolated from different sources, (bone marrow [BM-MSCs] and adipose tissue [ADSCs]), differ in their potential to regenerate smooth muscles in tissue-engineered urinary bladders and to determine an optimal number of MSCs for urinary bladder smooth muscle regeneration. Forty-eight rats underwent hemicystectomy and bladder augmentation with approximately 0.8 cm2 graft. In the first and second groups, urinary bladders were reconstructed with small intestinal submucosa (SIS) seeded with 10 × 106 or 4 × 106 ADSCs/cm2, respectively. In the third and fourth groups, urinary bladders were augmented with SIS seeded with 10 × 106 or 4 × 106 BM-MSCs/cm2, respectively. In the fifth group, urinary bladders were augmented with SIS without cells. The sixth group (control) was left intact. Smooth muscle regeneration was evaluated by real-time polymerase chain reaction (RT-PCR) and histological examinations. Histologically, there were no significant differences between urinary bladders augmented with ADSCs and BM-MSCs, but there was a marked increase in smooth muscle formation in bladders augmented with grafts seeded with MSCs in higher density (10 × 106/cm2) compared to lower density (4 × 106/cm2). Molecular analysis revealed that bladders reconstructed with ADSC-seeded grafts expressed higher levels of smooth muscle myosin heavy chain, caldesmon, and vinculin. Bladders augmented with unseeded SIS were fibrotic and devoid of smooth muscles. ADSCs and BM-MSCs have comparable smooth muscle regenerative potential, but the number of MSCs used for graft preparation significantly affects the smooth muscle content in tissue-engineered urinary bladders.

Transdifferentiation of Bone Marrow Mesenchymal Stem Cells into the Islet-Like Cells: the Role of Extracellular Matrix Proteins.

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 ß 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.

Long-term influence of bone marrow-derived mesenchymal stem cells on liver ischemia-reperfusion injury in a rat model.

BACKGROUND: The aim of this study was to evaluate the long-term usefulness of intraportal injection of the bone marrow-derived mesenchymal stem cells (BM-MSCs) in limitation of experimentally induced ischemia-reperfusion injury (IRI) in a rat model. MATERIAL AND METHODS: Twenty Wistar rats were divided into 3 groups: donor group (n=5), study group (n=10), and control group (n=5). IRI was performed using a modified hanging-weight system after left portal triad occlusion in study group animals. Isolated autologous BM-MSCs were labeled with fluorochrome PKH-26 then intraportally injected into the rats in the study group. Control group animals were intraportally injected with 1 ml of PBS. Follow-up was 3 months, after which animals were sacrificed for histopathological examination. Migration of BM-MSCs into different organs was examined. RESULTS: H&E staining of liver tissue sections from "time zero" biopsies did not show many irregularities in structural or histological construction compared to liver sections from the control group. However, a small amount of centrilobular hepatocyte necrosis and coagulative necrosis with neutrophil infiltration areas was observed in liver sections of the study group. The migration assay of BM-MSCs labeled with PKH-26 showed the highest positive BM-MSCs staining (6%) in the spleen, while few positively stained cells were found (2%) in liver sections. No BM-MSCs were detected in brain, kidney, or lung tissues. CONCLUSIONS: These results suggest that intraportal bone marrow-derived mesenchymal stem cell injection is safe and cells do not migrate chaotically to other organs after targeted implementation.

Is the poly (L- lactide- co- caprolactone) nanofibrous membrane suitable for urinary bladder regeneration?

The purpose of this study was to compare: a new five-layered poly (L-lactide-co-caprolactone) (PLC) membrane and small intestinal submucosa (SIS) as a control in rat urinary bladder wall regeneration. The five-layered poly (L-lactide-co-caprolactone) membrane was prepared by an electrospinning process. Adipose tissue was harvested from five 8-week old male Wistar rats. Adipose derived stem cells (ADSCs) were seeded in a density of 3×10(6) cells/cm2 onto PLC membrane and SIS scaffolds, and cultured for 5-7 days in the stem cell culture medium. Twenty male Wistar rats were randomly divided into five equal groups. Augmentation cystoplasty was performed in a previously created dome defect. Groups: (I) PLC+ 3×10(6)ADSCs; (II) SIS+ 3×10(6)ADSCs; (III) PLC; (IV) SIS; (V) control. Cystography was performed after three months. The reconstructed urinary bladders were evaluated in H&E and Masson's trichrome staining. Regeneration of all components of the normal urinary bladder wall was observed in bladders augmented with cell-seeded SIS matrices. The urinary bladders augmented with SIS matrices without cells showed fibrosis and graft contraction. Bladder augmentation with the PLC membrane led to numerous undesirable events including: bladder wall perforation, fistula or diverticula formation, and incorporation of the reconstructed wall into the bladder lumen. The new five-layered poly (L-lactide-co-caprolactone) membrane possesses poorer potential for regenerating the urinary bladder wall compared with SIS scaffold.

Filling effects, persistence, and safety of dermal fillers formulated with stem cells in an animal model.

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.

[The effect of fluoxetine and tianeptine on emotional and eating disorders in postmenopausal women]. / Wplyw fluoksetyny i tianeptyny na stan emocjonalny i zaburzenia odzywiania u kobiet po menopauzie.

UNLABELLED: In postmenopausal period, there occur a variety of psychosomatic symptoms in a female organism, which include deterioration of mood, anxiety, fear, sleep disorders, excessive irritability, impaired concentration and memory, overt depression with psychomotor retardation, decreased interest and life activity, decreased or increased appetite. Fluoxetine -selective serotonin reuptake inhibitor (SSRI) is used in the treatment of obesity and depression. Tianeptine--selective serotonin reuptake enhancer (SSRE) is used in the treatment of depressive episodes. Metabolic effects of prolonged tianeptine use are not well known. The aim of the study was the evaluation of the effect of long-term use of fluoxetine and tianeptine on emotional state and eating disorders in postmenopausal women. MATERIAL AND METHODS: The study was conducted in two groups of 30 postmenopausal women each, aged 52-66 years. The patients took fluoxetine for 6 months at a dose of 1 x 20 mg in the morning (group I) or tianeptine at a dose of 3 x 12.5 mg/daily (group II). Follow-up visits were conducted at 2, 4, 8, 12, 16 and 20 weeks. At week 24, extended tests were performed, including the assessment of the level of anxiety (Hamilton Anxiety Rating Scale-HARS) and depression (Beck Depression Inventory-BDI), the body mass index (BMI) and the waist/ hip ratio (WHR), which were compared with the corresponding results prior to the treatment with fluoxetine and tianeptine. RESULTS: After 6 months, in the group receiving fluoxetine, the reduction of the level of anxiety was obtained from 22.92 +/- 4.08 points to 12.36 +/- 2.43 points (p < 0.001) and a decrease in depression symptoms from 19.28 +/- 2.53 points to 10, 44 +/- 2.02 points (p < 0.001), but no reduction in body mass index was obtained -respectively 29.4 +/- 3.54 and 29.7 +/- 3.26 (p > 0.05) nor in the waist/ hip ratio--respectively 0.886 +/- 0.03 and 0.879 +/- 0.03 (p > 0.05). After 6 months, in the group receiving tianeptine, the reduction of the level of anxiety was achieved from 22.00 +/- 3.35 points to 15.20 +/- 3.42 points (p < 0.001) and a decrease in depression symptoms from 18.80 +/- 2.45 points to 14,16 +/- 4.06 points (p < 0.001), whereas no reduction in body mass index was obtained- respectively 27.7 +/- 2.81 and 27.4 +/- 2.65 (p > 0.05), even though slightly reduced waist/ hip ratio--from 0.877 +/- 0.03 to 0.863 +/- 0.03 (p < 0.05) was observed. Fluoxetine was more effective than tianeptine in the reduction of the level of anxiety (chi2 = 17.459, p < 0.01) as well as the reduction in the symptoms of depression (chi2 = 17.469, p < 0.01). CONCLUSIONS: Both tianeptine and fluoxetine are effective in the treatment of emotional disturbances in postmenopausal women. Both drugs may decrease appetite but long-term treatment does not alter body weight to the desired extent and they do not match the standards as monotherapy for obesity.

Is regenerative medicine a new hope for kidney replacement?

The availability of kidney and other organs from matching donors is not enough for many patients on demand for organ transplant. Unfortunately, this situation is not better despite the many of new interesting projects of promoting family, cross or domino transplants. These inexorable global statistics forced medical researchers to find a new potential therapeutic option that would guarantee safety and efficacy for the treatment of ESRD comparable to kidney transplantation. The aim of our review is to summarize the scientific literature that relating to the modern as well as innovative experimental methods and possibilities of kidney regeneration and, in addition, to find whether the regenerative medicine field will be a new hope for curing the patient with renal disease complications. The most important achievements in the field of regenerative medicine of kidney, which were mentioned and described here, are currently cumulated in 4 areas of interest: stem cell-based therapies, neo-kidneys with specially designed scaffolds or cell-seeded matrices, bioartificial kidneys and innovative nanotechnologically bioengineered solutions. Nowadays, we can add some remarks that the regenerative medicine is still insufficient to completely replace current therapy methods used in patients with chronic kidney disease especially with the end-stage renal disease where in many cases kidney transplantation is the only one chance. But we think that development of regenerative medicine especially in the last 20 years brings us more and more closer to solve many of today's problems at the frontier of nephrology and transplantology.

The effect of melatonin supplementation on the quality of sleep and weight status in postmenopausal women.

AIM OF THE STUDY: We evaluated the effect of melatonin supplementation on the nutritional status of postmenopausal women. MATERIAL AND METHODS: The study included 56 women (51-65 years) and 25 healthy women (27-36 years). The emotional state was assessed using Hamilton Depression Rating Scale (HAM-D), the quality of sleep using Insomnia Severity Index (ISI). Body mass index (BMI) and waist-hip ratio (WHR) were also calculated. The patients were divided into 3 groups: group I (control) - 25 women with normal body weight, group II - 26 postmenopausal women with normal body weight, group III - 30 postmenopausal women with high body weight. In women from group II and III, routine laboratory tests, levels of thyroid-stimulating hormone (TSH), 17ß-estradiol, prolactin, follicle-stimulating hormone (FSH) and the concentration of 6-hydroxymelatonin sulphate (6-HMS) in day/night urine fractions were determined. On the day of the examination, women remained on a liquid diet (1800 kcal). Next, a balanced diet of 1500 kcal and 5 mg of melatonin administration were recommended. The follow-up examinations were performed after 4, 8, 12, 16, 20 and 24 weeks. RESULTS: The patients from groups II and III showed similar mild levels of anxiety and depression and a significant degree of sleep disorders. In group III, lower urinary 6-HMS excretion was observed at night. In both groups a negative correlation was found between urinary 6-HMS excretion and the degree of sleep disorders. After 24 weeks, a statistically significant improvement of quality of sleep was obtained. A negative correlation was detected between urinary 6-HMS excretion and BMI. CONCLUSION: Melatonin supplementation contributed to body weight reduction.