Adipose Tissue-derived Stem Cell Therapy Research Articles

Application of adipose tissue-derived stem cell therapy with a clinically relevant dose does not significantly affect atherosclerotic plaque characteristics in a streptozotocin-induced hyperglycaemia mouse model

AimsDiabetes mellitus (DM) induces increased inflammation of atherosclerotic plaques, resulting in elevated plaque instability. Mesenchymal stem cell (MSC) therapy was shown to decrease plaque size and increase stability in non-DM animal models. We now studied the effect of MSC therapy in a streptozotocin-induced hyperglycaemia mouse model using a clinically relevant dose of adipose tissue-derived MSCs (ASCs). MethodsHyperglycaemia was induced in male C57BL/6 ApoE−/− mice (n=24) via intraperitoneal streptozotocin (STZ) injection (0.05 mg/g bodyweight) for 5 consecutive days. 16 weeks after the first STZ injection, the mice received either 100,000 ASCs (n=9) or vehicle (n=14) intravenously. The effects of ASC treatment on the size and stability of aortic root atherosclerotic plaques were determined 4 weeks post-treatment via (immuno)histochemical analyses. Furthermore, plasma monocyte subsets within 3 days pre- and 3 days post-treatment, and 4 weeks post-treatment, were studied. ResultsASC treatment did not significantly affect atherosclerotic plaque size or intra-plaque inflammation. Although ASC-treated mice had a higher percentage of intra-plaque fibrosis (42.5±3.3%) compared to vehicle-treated mice (37.6±6.8%, p=0.07), this did not reach significance. Additionally, although differences in the percentages of circulating pro- and anti-inflammatory monocytes were observed after ASC treatment compared to pre-treatment (p=0.005), their levels did not differ significantly at any time point compared to vehicle-treated mice. ConclusionsASC treatment with a clinically relevant dose did not significantly affect atherosclerotic plaque size or intra-plaque inflammation in a hyperglycaemia mouse model. Despite a borderline significant improvement in intraplaque fibrotic content, the potential of ASC treatment on atherosclerotic plaque stability in a diabetic environment remains to be determined.

Seven-Year Longitudinal Study: Clinical Evaluation of Knee Osteoarthritic Patients Treated with Mesenchymal Stem Cells.

Background/Objectives: Numerous studies have demonstrated the safety and efficacy of intraarticular stem cell injections for treating osteoarthritic knee joints, reporting symptom reduction and pain relief within a few months of treatment. Here, we report the results of a 7-year follow-up after a single intraarticular injection of 0.5-1 × 107 autologous adipose tissue-derived mesenchymal stem cells in patients with OA (Kellgren-Lawrence grade 2 to 4). Methods: Nine patients were treated, and two patients had bilateral disease. Patients were evaluated clinically and radiologically using X-ray and MRI. A comprehensive statistical analysis was undertaken to evaluate the obtained results. Results: All clinical scores and range of motion significantly improved within the first six months after injection. At the 18-month time point, a significant improvement in cartilage structure was observed on MRI while X-ray showed no changes in subchondral bone of distal femur and proximal tibia. At the 60-month time point, the clinical scores were still improved compared to baseline, except for the range of motion, which decreased almost back to the baseline level. At 84 months, the clinical scores decreased significantly toward the baseline level, but the MRI structural characteristics of cartilage still remained significantly better than those measured at baseline. Conclusions: Adipose tissue-derived stem cell therapy has substantial long-term clinical effects on patients with knee osteoarthritis.

Effect of adipose tissue-derived stem cells therapy on clinical response in patients with primary Sjogren’s syndrome

The purpose of this trial was to clinically assess the effect and safety of Adipose Tissue-derived Stem Cells (ADSCs) treatment on primary Sjogren’s Syndrome (pSS). In this 6-month randomized, triple-blind, placebo-controlled clinical trial, pSS patients were randomly assigned to two groups. After demographic characteristics and clinical examination were achieved, local injection of ADSCs into bilateral glands was performed with patients in ADSCs group (n = 35) and placebo solution was used for another group (n = 39) at three time points. Patients were followed up at 1-, 3- and 6-month. At each visit, studies of clinical and laboratory outcomes, as well as subjective symptoms, were conducted. A total of 74 subjects who met the including criteria were allocated in two groups and eventually 64 subjects (86.5%) completed the treatments and the follow-up assessments. Secretion of salivary and lachrymal glands were significantly improved in 3-month (P < 0.05). A great improvement of European League Against Rheumatism Sjögren’s Syndrome Disease Activity Index (ESSDAI) was found after ADSCs treatment with intergroup comparison from baseline to follow-up (P < 0.05). There is also a significant difference of European Alliance of Associations for Rheumatology SS Patient Reported Index (ESSPRI) between the two groups in the follow-up (P < 0.05). A significant abatement of IgG, IgM, C3, C4 and ESR between two groups was observed in part of follow-up time points (P < 0.05). The ADSCs therapy can provide relief of oral and eye’s dryness in our trial in a short time and has potential improvement of subjective and systemic syndromes of pSS.

"Adipose-derived mesenchymal stem cell therapy for the management of female sexual dysfunction: Literature reviews and study design of a clinical trial".

Hormone imbalance and female sexual dysfunction immensely affect perimenopausal female health and quality of life. Hormone therapy can improve female hormone deficiency, but long-term use increases the risk of cardiovascular diseases and cancer. Therefore, it is necessary to develop a novel effective treatment to achieve long-term improvement in female general and sexual health. This study reviewed factors affecting syndromes of female sexual dysfunction and its current therapy options. Next, the authors introduced research data on mesenchymal stromal cell/mesenchymal stem cell (MSC) therapy to treat female reproductive diseases, including Asherman’s syndrome, premature ovarian failure/primary ovarian insufficiency, and vaginal atrophy. Among adult tissue-derived MSCs, adipose tissue-derived stem cells (ASCs) have emerged as the most potent therapeutic cell therapy due to their abundant presence in the stromal vascular fraction of fat, high proliferation capacity, superior immunomodulation, and strong secretion profile of regenerative factors. Potential mechanisms and side effects of ASCs for the treatment of female sexual dysfunction will be discussed. Our phase I clinical trial has demonstrated the safety of autologous ASC therapy for women and men with sexual hormone deficiency. We designed the first randomized controlled crossover phase II trial to investigate the safety and efficacy of autologous ASCs to treat female sexual dysfunction in perimenopausal women. Here, we introduce the rationale, trial design, and methodology of this clinical study. Because aging and metabolic diseases negatively impact the bioactivity of adult-derived MSCs, this study will use ASCs cultured in physiological oxygen tension (5%) to cope with these challenges. A total of 130 perimenopausal women with sexual dysfunction will receive two intravenous infusions of autologous ASCs in a crossover design. The aims of the proposed study are to evaluate 1) the safety of cell infusion based on the frequency and severity of adverse events/serious adverse events during infusion and follow-up and 2) improvements in female sexual function assessed by the Female Sexual Function Index (FSFI), the Utian Quality of Life Scale (UQOL), and the levels of follicle-stimulating hormone (FSH) and estradiol. In addition, cellular aging biomarkers, including plasminogen activator inhibitor-1 (PAI-1), p16 and p21 expression in T cells and the inflammatory cytokine profile, will also be characterized. Overall, this study will provide essential insights into the effects and potential mechanisms of ASC therapy for perimenopausal women with sexual dysfunction. It also suggests direction and design strategies for future research.

Efficacy and safety of autologous adipose tissue-derived stem cell therapy for children with refractory Crohn's complex fistula: a Phase IV clinical study.

PurposeAutologous adipose tissue-derived stem cells (ASCs) have been proposed for patients with refractory Crohn disease, but research is lacking in pediatric patients. This Phase IV study evaluated the efficacy and safety of ASCs in children with refractory Crohn's fistulae.MethodsPatients with a refractory Crohn's fistula who did not have conventional therapy for more than 3 months or with a recurrent complex Crohn's fistula were included. All patients were at least 14 years old. Patients with infection, poor condition, or active Crohn disease with a disease activity index of 450 and above were excluded. Five patients were treated with ASCs from 2014 to 2015 in Asan Medical Center. ASC administration was adjusted according to fistula size (1 mL per cm2). We evaluated the efficacy and safety 8 weeks after injection and followed patients for 6 months.ResultsFistulae were healed in 4 patients by 8 weeks after ASC injection. Of these 4 patients, 1 had complete fistula closure and sustainability after 6 months. The other 3 with healing effects had less than 50% fistula closure by 6 months. None of these 4 patients have persistent fistulae. One patient had no healing effect, and seton ligation was performed 8 months after ASC injection. There were no adverse effects related to ASC administration.ConclusionASC therapy is a simple and well-tolerated therapeutic option for children with refractory Crohn's complex fistulae. Complete closure was well-sustained. However, more data from a larger number of patients are needed.

Liver damage in schistosomiasis is reduced by adipose tissue-derived stem cell therapy after praziquanteltreatment.

BackgroundIn view of the potential immunosuppressive and regenerative properties of mesenchymal stem cells (MSC), we investigated whether transplantation of adipose tissue-derived stem cells (ASC) could be used to control the granulomatous reaction in the liver of mice infected with Schistosoma mansoni after Praziquantel (PZQ) treatment.Methodology/Prinicpal findingsC57BL/6 mice infected with S. mansoni were treated with PZQ and transplanted intravenously with ASC from uninfected mice. Liver morpho-physiological and immunological analyses were performed. The combined PZQ/ASC therapy significantly reduced the volume of hepatic granulomas, as well as liver damage as measured by ALT levels. We also observed that ASC accelerated the progression of the granulomatous inflammation to the advanced/curative phase. The faster healing interfered with the expression of CD28 and CTLA-4 molecules in CD4+ T lymphocytes, and the levels of IL-10 and IL-17 cytokines, mainly in the livers of PZQ/ASC-treated mice.ConclusionsOur results show that ASC therapy after PZQ treatment results in smaller granulomas with little tissue damage, suggesting the potential of ASC for the development of novel therapeutic approaches to minimize hepatic lesions as well as a granulomatous reaction following S. mansoni infection. Further studies using the chronic model of schistosomiasis are required to corroborate the therapeutic use of ASC for schistosomiasis.

Effect of Intracorporeal Human Adipose-Derived Stem Cells (hADSCs) on Corpora Cavernosa Transforming Growth Factor β1 (TGFβ1) and Collagen Type I Concentration in Wistar Rat Priapism Model.

IntroductionThe ischemic process in priapism can lead to displacement of normal tissue with fibrotic tissue, due to collagen deposition, and eventually leads to erectile dysfunction. Many studies have identified that the supernatant of adipose tissue–derived stem cells (ADSCs) significantly ameliorates fibrosis of different tissue, but limited attention has been paid to its efficacy on fibrosis of the corpora cavernosa.MethodsA total of 22 Wistar rats divided into five groups, with two groups each consisting of five male wistar rats with priapism without human ADSC (hADSC) therapy (group I) and two other groups consisting of five rats with priapism, were given 106 cells' intracorporeal hADSC injection after 12 hours of penile clamping (group II) were euthanized after 2 and 4 weeks of observation. The last group consisted of two rats without any treatment or model (group III). Following euthanasia, penises were harvested for TGFβ1 and collagen type I measurement using ELISA. Statistical analysis using independent-sample t-tests was done with SPSS 21.0.ResultsPenile TGFβ1 concentration in the treatment group was significantly lower in the second and fourth weeks of observation (p2=0.004, p4=0.003), and collagen type I was significantly lower in the second and fourth weeks (p2=0.003, p4=0.011).ConclusionIntracorporeal hADSC injection limited the fibrosis process in a priapism model. Although the mechanism was unclear, it may be related to the potential of hADSCs to produce various growth factors that could limit TGFβ1 and collagen production.

Autologous Stem Cell Therapy for Cerebral Palsy

Introduction: We describe treatment of Cerebral Palsy with adult stem cells derived from bone marrow and fat of the same patient. Adult stem cells are of two types, the mesenchymal and haemopoietic stem cells which have potential to duplicate, indefinitely produce 50 types of growth factors that repair and regenerate tissues in an epigenetic manner. Every organ has its own stem cells, for example kidney stem cells, liver stem cells, etc. When specialized cells in an organ get damaged, the local stem cells come forward and get differentiated into specialized cells and the tissue damage is replenished. But when the stock of this reserve of local stem cell is over, the organ starts failing. In autologous stem cell therapy, we harvest stem cells from other healthy organs like fat and bone marrow which have abundant stem cells and put them into the diseased organ. Adult stem cells differentiate into neurons in vitro when added with nerve growth factor which is present in every nerve tissue. Our stem cell research was done by following all the guidelines set at national and international level. In India the incidence of cerebral palsy is around 5/1000 live births. In cerebral Palsy the upper motor neuron is weak, hence the lower motor neuron becomes hyper active leading to spasticity. The treatment involves mainly physiotherapy and prognosis is poor. Physiotherapy is a palliation and does not repair damage in the brain. The conventional therapy has failed to give any satisfactory results in these children hence something new needs to be done. Materials and Methods: In last 2 years we did stem cell therapy for 21 cases of CP in our center with age of 9 months to 17 years. Out of 21 cases of CP, 7 (33.3%) cases had quadriplegia, 6 (28.57%) paraplegia and 1 (4.76%) monoplegia, 2 (9%) hypotonic, 5 (23.8%) hemiplegia, 4 (19.04%) with mental retardation and 3 (14.28) had features of ADHD and Autism. 18 patients had squint in the eyes, 8 patients had food regurgitation and drooling of saliva, 2 patients had audiovisual impairment, 3 patients had bronchial asthma and one patient had menstrual irregularity. 26 cases were put as control with age range of 9 months to 19 years. 9 (34.61%) patients had quadriplegia, 8 (30.76%) paraplegia and 3 (11.53) had monoplegia, 3 (11.53%) hypotonic, 5 (19.23) with mental retardation and 6 (23.07) had features of ADHD and autism. 21 patients had squint in the eyes, 11 patients had food regurgitation and drooling of saliva, 4 patients had audiovisual impairment, 5 patients had bronchial asthma in the control group. 11 patients were subjected to adipose tissue derived stem cell therapy and 10 with bone marrow derived stem cell therapy. All patients were examined at 3 monthly intervals. Maximum follow up was 2.5 years and minimum of one year. Results: The results of bone marrow derived stem cells and adipose tissue derived stem cells were almost the same. Results took 3 months to appear and positive outcome came till one year since therapy. The spasticity was significantly reduced in 15 out of 18 patients in 6 months. Out of 18 patients who had squint, it was regressed completely in 12 (66.66%) patients and partially in 3 (16.66%) in 6 months’ time. 8 patients had faulty deglutition and 75% of them had 90% reg

Adipose Tissue-Derived Stem Cell Therapy for Cavernous Nerve Injury-Induced Erectile Dysfunction in the Rat Model: A Systematic Review and Meta-Analysis Using Methodological Quality Assessment.

Background and ObjectivesFew studies were evaluated the effect of blindness on outcome in animal models, though a potential effect of blinding has been reported in clinical trials. We evaluated the effects of adipose tissue-derived stem cells (ADSCs) on cavernous nerve injury (CNI)-induced erectile dysfunction (ED) in the rat and examined how proper blinding of the outcome assessor affected treatment effect.Methods and ResultsWe searched in Pubmed, EMBASE, Cochrane and Web of Science databases from inception to January 2019. We included CNI animal model, randomized controlled experiments, and ADSC intervention. Erectile function and structural changes were assessed by intracavernous pressure and mean arterial pressure (ICP/MAP) ratios, neuronal nitric oxide synthase (nNOS) levels, cavernous smooth muscle and collagen (CSM/collagen) ratios, and cyclic guanosine monophosphate (cGMP).ResultsNineteen studies were included in the final meta-analysis. The ICP/MAP ratio of the ADSC treatment group increased compared to the control group (SMD=1.33, 95%CI: 1.11~1.56, I2=72%). The nNOS level (SMD=2.29, 95%CI: 1.74~2.84, I2=75%), CSM/collagen (SMD=2.57, 95%CI: 1.62~3.52; I2=85%), and cGMP (SMD=2.96, 95%CI: 1.82~4.10, I2=62%) were also increased in the ADSC treatment group. Preplanned subgroup analysis was conducted to explore the source of heterogeneity. Five studies with blinded outcome assessment were significantly less effective than the unblinded studies (SMD=1.33, 95%CI: 0.86~1.80; SMD=1.81, 95%CI: 1.17~2.46, respectively).ConclusionsADSCs might be effective in improving erectile function and structural change in CNI-induced ED. However, non-blinded outcome assessors might cause detection bias and overestimate treatment efficacy. Therefore, the ADSC efficacy must be further evaluated with a rigorous study design to avoid bias.

Antioxidant preconditioning enhances adipose tissue-derived stem cell therapy by increasing intrahepatic engraftment for liver fibrosis

Introduction: Although it has been pre-clinically suggested that adipose-derived mesenchymal stem cells (ADSC)-based therapy can effectively treat liver cirrhosis, the hepatic engraftment of ADSC is still extremely low, which affects the clinical translation of ADSC for liver cirrhosis treatment. Method: ADSCs were treated with hydrogen peroxide to induce cell stress. Antioxidants, including reduced glutathione and melatonin were co-treated to examine their protective effects and mechanisms on stem cell damage. Liver fibrosis model was also established to test the transplantation efficacy of ADSCs with or without antioxidants pre-incubation. Result: Our data showed that precondition of antioxidants synergistically enhances the anti-stress ability of ADSCs induced by hydrogen peroxide in vitro. We found that preconditioning with antioxidants significantly improves therapeutic efficacy of ADSCs transplantation in liver fibrosis mice, including ameliorating histological damage, oxidative stress, inflammation and fibrosis. In addition, we also found that preconditioned with antioxidants increases intrahepatic engraftment of ADSCs through reducing the hepatic content of NOXs and ROS both in vitro and in vivo. Conclusion: These findings suggested that antioxidant preconditioning can enhance ADSC-based therapy for liver fibrosis via in increasing hepatic engraftment of ADSCs, which may through suppressing NOXs/ROS signaling.

Clinical assessment after human adipose stem cell transplantation into dogs.

Adipose tissue-derived stem cell (ASCs) are an attractive source of stem cells with therapeutic applicability in various fields for regenerating damaged tissues because of their stemness characteristics. However, little has reported on evaluating adverse responses caused by human ASC therapy. Therefore, in the present study, a clinical assessment after human ASC transplantation into dogs was undertaken. A total of 12 healthy male dogs were selected and divided into four groups: saline infusion, saline bolus, ASC infusion, and ASC bolus groups. Physical assessment and blood analysis were performed following ASC transplantation, and the concentrations of angiogenic factors, and pro- and anti-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA). There were no adverse vital sign responses among the dogs. Blood analyses revealed no remarkable complete blood count or serum chemistry results. ELISA results for angiogenic and anti-inflammatory factors including matrix metalloproteinase 9 (MMP9), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and interleukin-10 (IL-10) were significantly higher in the two ASCs groups than in the controls. In conclusion, this study demonstrated that transplantation of human ASCs produced no adverse effects and could be used safely in dogs. In addition, human ASCs could be involved in modulating secretions of angiogenic factors including MMP9, VEGF, bFGF, and HGF and anti-inflammatory factor IL-10.

Adipose-Derived Stem Cells Ameliorate Experimental Murine Colitis via TSP-1-Dependent Activation of Latent TGF-β.

Adipose tissue-derived stem cells (ASCs) have been investigated as therapeutic tools for a variety of autoimmune diseases, including inflammatory diseases. However, the mechanisms underlying the immunomodulatory properties of ASCs are not well understood. Here, we investigated the mechanism of regulatory T cell (Treg) induction in ASC therapy in a murine model of inflammatory bowel disease. Acute colitis was induced in mice using dextran sulfate sodium and ASCs administered intraperitoneally. Tregs and CD103+ dendritic cells were analyzed in the mesenteric lymph nodes (MLNs), spleen, and colonic lamina propria (CLP). Activation of latent TGF-β by ASCs was analyzed in vitro using ELISA. siRNA technology was used to create ASCs in which TSP-1 or integrinαv was knocked down in order to investigate the involvement of these proteins in the activation of latent TGF-β. In addition, TSP-1-knockdown ASCs were administered to mice with colitis to assess their clinical efficacy in vivo. Systemic administration of ASCs significantly lessened the clinical and histopathological severity of colitis. ASCs were distributed throughout the lymphatic system in the MLNs and spleen. Tregs were increased in the MLNs and CLP, but CD103+ dendritic cells were not significantly altered. The ASCs activated latent TGF-β. TSP-1 knockdown impaired TGF-β activation in vitro and abrogated the therapeutic effects of the ASCs in vivo. Furthermore, Tregs were not increased in the MLNs and CLP from mice treated with TSP-1-knockdown ASCs. These results demonstrate that ASCs induce Tregs by activating latent TGF-β via TSP-1, independent of CD103+ dendritic cell induction.

Adipose tissue-derived stem cell therapy for erectile dysfunction in rats: a systematic review and meta-analysis.

We aimed to systematically assess the effect of adipose tissue-derived stem cell (ADSC) therapy and its influential factors on the treatment of erectile dysfunction (ED) in rats. Two authors independently searched for published studies through PubMed and EMBASE from study inception until August 31, 2016. A meta-analysis was used to combine the effect estimate from the published studies. A subgroup analysis was performed to identify the effect of some influential factors. The pooled standardized mean differences (SMDs) with 95% confidence intervals (CIs) were calculated by a fixed-effects or random-effects model analysis. Twenty studies with a total of 248 rats were included in this meta-analysis. The pooled analysis showed that ADSC therapy significantly increased the ratio of intracavernous pressure and mean arterial pressure (ICP/MAP; SMD 3.46, 95% CI 2.85-4.06; P<0.001) compared to control therapy. The levels of neuronal nitric oxide synthase (nNOS; SMD 6.37, 95% CI 4.35-8.39; P<0.001), the cavernous smooth muscle content (CSMC; SMD 3.65, 95% CI 2.65-4.65; P<0.001), the ratio of cavernous smooth muscle and collagen (CSM/collagen; SMD 4.16, 95% CI 2.59-5.72; P<0.001), and the cyclic guanosine monophosphate (cGMP; SMD 7.12, 95% CI 2.76-11.48; P=0.001) were higher following ADSC therapy than following control therapy. Subgroup analysis showed that ADSCs modified by growth or neurotrophic factors significantly recovered erectile function (P<0.001) compared with ADSC therapy. The adequate data indicated that ADSC therapy recovered erectile function and regenerated cavernous structures in ED rats, and ADSCs modified by some growth and neurotrophic factors accelerated the recovery of erectile function and cavernous structures in ED rats.

Superparamagnetic iron oxide nanoparticle targeting of adipose tissue-derived stem cells in diabetes-associated erectile dysfunction.

Erectile dysfunction (ED) is a major complication of diabetes, and many diabetic men with ED are refractory to common ED therapies. Adipose tissue-derived stem cells (ADSCs) have been shown to improve erectile function in diabetic animal models. However, inadequate cell homing to damaged sites has limited their efficacy. Therefore, we explored the effect of ADSCs labeled with superparamagnetic iron oxide nanoparticles (SPIONs) on improving the erectile function of streptozotocin-induced diabetic rats with an external magnetic field. We found that SPIONs effectively incorporated into ADSCs and did not exert any negative effects on stem cell properties. Magnetic targeting of ADSCs contributed to long-term cell retention in the corpus cavernosum and improved the erectile function of diabetic rats compared with ADSC injection alone. In addition, the paracrine effect of ADSCs appeared to play the major role in functional and structural recovery. Accordingly, magnetic field-guided ADSC therapy is an effective approach for diabetes-associated ED therapy.

Fat for fostering: Regenerating injured heart using local adipose tissue

Fat for fostering: Regenerating injured heart using local adipose tissue

State of the art. Autologous fat graft and adipose tissue-derived stromal vascular fraction injection for hand therapy in systemic sclerosis patients

Systemic sclerosis is an autoimmune disease characterized by sclerosis (hardening) of the skin and deep viscera associated with microvascular functional and structural alteration, which leads to chronic ischemia. In the hands of patients, ischemic and fibrotic damages lead to both pain and functional impairment. Hand disability creates a large burden in professional and daily activities, with social and psychological consequences. Currently, the proposed therapeutic options for hands rely mainly on hygienic measures, vasodilatator drugs and physiotherapy, but have many constraints and limited effects. Developing an innovative therapeutic approach is crucial to reduce symptoms and improve the quality of life. The discovery of adult stem cells from adipose tissue has increased the interest to use adipose tissue in plastic and regenerative surgery. Prepared as freshly isolated cells for immediate autologous transplantation, adipose tissue-derived stem cell therapy has emerged as a therapeutic alternative for the regeneration and repair of damaged tissues. We aim to update literature in the interest of autologous fat graft or adipose derived from stromal vascular fraction cell-based therapy for the hands of patients who suffer from systemic sclerosis.

Cytokines inflammatory genes profile in experimental frozen-thawed ovarian grafts treated with scaffold-base delivery of adipose tissue-derived stem cells

Event Abstract Back to Event Cytokines inflammatory genes profile in experimental frozen-thawed ovarian grafts treated with scaffold-base delivery of adipose tissue-derived stem cells Luciana Damous1, Juliana S. Nakamuta2, Ana E. Saturi De Carvalho2, Katia C. Carvalho1, Manuel J. Simões3, José M. Soares-Jr1, José E. Krieger2 and Edmund C. Baracat1 1 Faculdade de Medicina da Universidade de São Paulo, Obstetrics and Gynecology, Brazil 2 Instituto do coração, Faculdade de Medicina da Universidade de São Paulo, Laboratório de Cardiologia Genética e Molecular, Brazil 3 Universidade Federal de São Paulo, Morphology, Brazil Introduction: Previous works of our group demonstrated that cellular therapy with adipose tissue-derived stem cells (ASC) of frozen-thawed ovarian autografts preserves the graft morphology and didn’t enhance fibrosis[1]. However, new researchers are necessary to better evaluate the inflammatory response in this graft. Materials and Methods: Twelve 12-week-old adult female Wistar rats were use. Frozen-thawed ovarian grafts of adult female rats were treated with rat ASC delivery in an acellular matrix (Gelfoam) immediately after an autologous retroperitoneal transplant (n=6). Controls received Gelfoam with vehicle (DMEM low glucose, n=6). The ovarian grafts were retrieved 30 days after transplantation. Quantitative gene expression (qPCR) for inflammatory cytokines (84 genes) was evaluated by Real Time-Polymerase Chain Reaction (RT-PCR). All reactions were performed in triplicate and controlled by negative RT (no enzyme) and no-template controls and all gene expression levels were normalized to the mean of the internal genes. A Fold Regulation reference value of -2 and +2 was considered for expression analyses. Further analyses were done by Ingenuity Pathway Analysis (IPAÒ) software in order to determine the main canoninal pathways and the expression values of the top molecules. Results and Discussion: There were 36 genes up-regulated and three down-regulated. The ten top molecules up-regulated, in descending order, were CXCL6 (+13.1), IL21 (+10), LTA (+7.6), IL1A (+7.1), CXCL11 (+6.7), CCL19 (+6.4), CCR3 (+6.2), CCR6 (+5.4), LTB (+5.1) and CXCR2 (+5). The top canonical pathways involved were Differential Regulation of Cytokine Production in Macrophages and T Helper Cells (44%), Differential Regulation of Cytokine Production in Intestinal Epithelial Cells (43%), Role of Cytokines in Mediating Communication between Immune Cells (18%), Granulocyte Adhesion and Diapedesis (9%) and Agranulocyte Adhesion and Diapedesis (7%). The top diseases and biofunctions envolved were Inflammatory Response (38 molecules), Inflammatory Disease (37), Immunological Disease (37), Connective Tissue Disorders (31) and Skeletal and Muscular Disorders (31). Tox functions were little involved and represented mainly by hepatic mechanisms such as Increases Liver Damage (11 molecules), Hepatic Cholestasis (10), Hepatic Fibrosis (9), Liver Proliferation (9) and Increases Liver Hepatitis (6). Conclusion: ASC therapy based on scaffold base-delivery strategy in rat ovarian grafts induces upregulation in 36 genes involved in inflammatory cytokines pathway. New researchers are necessary to evaluate if these findings could compromise the graft quality. São Paulo Research Foundation (Process numbers 2010/17897-5 and 2012/09469-9); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for post-doctoral scholarship

Does adipose tissue-derived stem cell therapy improve graft quality in freshly grafted ovaries?

BackgroundA major concern in ovarian transplants is substantial follicle loss during the initial period of hypoxia. Adipose tissue-derived stem cells (ASCs) have been employed to improve angiogenesis when injected into ischemic tissue. This study evaluated the safety and efficacy of adipose tissue-derived stem cells (ASCs) therapy in the freshly grafted ovaries 30 days after injection.MethodsRat ASCs (rASCs) obtained from transgenic rats expressing green fluorescent protein (GFP)-(5 × 104 cells/ovary) were injected in topic (intact) or freshly grafted ovaries of 30 twelve-week-old adult female Wistar rats. The whole ovary was grafted in the retroperitoneum without vascular anastomosis, immediately after oophorectomy. Vaginal smears were performed daily to assess the resumption of the estrous cycle. Estradiol levels, grafts morphology and follicular viability and density were analyzed. Immunohistochemistry assays were conducted to identify and quantify rASC-GFP+, VEGF tissue expression, apoptosis (cleaved caspase-3 and TUNEL), and cell proliferation (Ki-67). Quantitative gene expression (qPCR) for VEGF-A, Bcl2, EGF and TGF-β1 was evaluated using RT-PCR and a double labeling immunofluorescence assay for GFP and Von Willebrand Factor (VWF) was performed.ResultsGrafted ovaries treated with rASC-GFP+ exhibited earlier resumption of the estrous phase (p < 0.05), increased VEGF-A expression (11-fold in grafted ovaries and 5-fold in topic ovaries vs. control) and an increased number of blood vessels (p < 0.05) in ovarian tissue without leading to apoptosis or cellular proliferation (p > 0.05). Estradiol levels were similar among groups (p > 0.05). rASC-GFP+ were observed in similar quantities in the topic and grafted ovaries (p > 0.05), and double-labeling for GFP and vWF was observed in both injected groups.ConclusionrASC therapy in autologous freshly ovarian grafts could be feasible and safe, induces earlier resumption of the estrous phase and enhances blood vessels in rats. This pilot study may be useful in the future for new researches on frozen-thawed ovarian tissue.Electronic supplementary materialThe online version of this article (doi:10.1186/s12958-015-0104-2) contains supplementary material, which is available to authorized users.

Scaffold-based delivery of adipose tissue-derived stem cells in rat frozen-thawed ovarian autografts: preliminary studies in a rat model.

This study aimed to evaluate whether a gelatin-based Gelfoam sponge is feasible as a scaffold for adipose tissue-derived stem cell (ASC) therapy in rat frozen-thawed ovarian autografts. Two sets of studies were performed. The in vitro set evaluated ASCs' viability in the Gelfoam scaffold at different times of co-culturing (after 24, 48, 72, 96, and 120 h). The in vivo set used 20 12-week-old adult female Wistar rats. Frozen-thawed ovarian grafts were treated with ASCs delivered in Gelfoam scaffolds immediately after an autologous retroperitoneal transplant (ASCs-GS, n = 10). The controls received Gelfoam with a culture medium (GS, n = 10). Assessment of graft quality was conducted by vaginal smears (until euthanasia on the 30th postoperative day), histological analyses, follicular density, and viability and fibrosis. Immunohistochemical staining for VEGF-A expression, vascular network (vWF), apoptosis (caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)), cell proliferation (Ki-67), and hormone receptors (estrogen and progesterone) were performed. The cells remained viable in Gelfoam for up to 120 h of co-culturing. The graft morphology was similar among the groups. ASC therapy promoted the earlier resumption of the estrous phase (GS 16.6 ± 3 vs. ASCs-GS 12.8 ± 1.3 days) and enhanced estrogen receptors compared with the controls (p < 0.05) without interfering with the quantity and viability of the ovarian follicles, fibrosis, endothelial cells, VEGF immunoexpression, apoptosis, or cell proliferation (p > 0.05). The Gelfoam scaffold could be a feasible and safe non-invasive technique for ASC delivery in the treatment of frozen-thawed ovarian autografts. Future studies should evaluate the real benefit of this treatment on the survival and endocrine activity of the graft.

Functional Recovery and in Vivo Cell Tracking after Human Adipose Tissue-Derived Stem Cells Therapy for Experimental Spinal Cord Injury

Introduction The use of human adipose tissue-derived stem cells (hADSC) as therapy in spinal cord injury (SCI) is a promising approach for the recovery of motor function of injured patients. This study aims to evaluate the effects of hADSC, applied to the cauda equina in an animal model of blunt spinal trauma, on their motor function. A secondary aim is to analyze cell migration toward the injury site. Method All samples were collected after an informed consent had been obtained in accordance with guidelines for research involving human subjects, and with the approval of the ethics committee (approval number 769). A total of 40 Wistar rats were anesthetized, injured at Th10 using the NYU impactor device and randomly divided into four groups: Group A, animals were submitted to one culture medium injection immediately after injury; group B, animals were submitted to two culture medium injections, immediately after and 7 days postlesion; group C, animals were submitted to one cell injection (1 × 106 cells) immediately after injury and group D, animals were submitted to two cell injections, immediately after and 7 days postlesion. Standard animal care, according to the university's protocol was maintained for 40 days with motor analysis every 72 hours using the Basso, Beattie, and Bresnahan (BBB) scale. Some animals were transplanted with luc gene-transduced cells to evaluate cell migration, which was assessed by in vivo bioluminescence images of animals in groups C and D (IVIS Lumina II). Results A statistically significant recovery of motor function was observed in animals transplanted with hADSC compared with the control group ( p = 0.007 for one injection protocol, p = 0.005 for the two injection protocol). No differences were observed between one and two injection protocols ( p = 0.13). Bioluminescence analysis revealed cell migration to the injury site after injection. Conclusion hADSC injected at the cauda equina migrate toward the injury site and promote significant motor recovery.