Human Adipose Tissue-derived Stem Cells Research Articles

The effect of polycaprolactone/graphene oxide electrospun scaffolds on the neurogenic behavior of adipose stem cells

Stem cell destiny can be controlled with scaffold biomaterials in tissue engineering and regenerative medicine. This study aimed to investigate the neuronal differentiation potential of human adipose tissue-derived mesenchymal stem cells in graphene nanofiber matrix in vitro. Stem cell isolation was performed from adipose tissue taken from human by mechanical and enzymatic methods. The differentiation potential was examined after incubation of adipose stem cells in normal medium and neural differentiation medium, on graphene oxide (GO) and polycaprolactone (PCL) composite scaffolds produced by electrospinning technique. In vitro studies indicated that the presence of GO in PCL scaffold increases an effect on cell attachment, proliferation, infiltration into the scaffold, and neuronal differentiation. Also, unlike subcutaneous tissue, it has been shown immunohistochemically that mesenchymal stem cells derived from epidural adipose tissue tend to differentiate into oligodendrocytes.

In Vitro Effect of Methamphetamine on Proliferation, Differentiation and Apoptosis of Adipose Tissue Stem Cells.

Among abused substances, methamphetamine is a psychostimulant drug widely used recreationally with public health importance. This study investigated the effect of methamphetamine on proliferation, differentiation, and apoptosis of human adipose tissue stem cells (AdSCs). AdSCs were isolated from human abdominal adipose tissue and were characterized for mesenchymal properties and growth kinetics. MTT assay was undertaken to assess methamphetamine toxicity on proliferation and differentiation properties and apoptosis of hAdSCs. Isolated cells were shown to have mesenchymal properties and a population doubling time (PDT) of 40.1 h. Following methamphetamine treatment, expressions of KI-67 and TPX2 as proliferation genes and Col1A1 and PPARg as differentiation genes decreased. Methamphetamine administration increased the expression of Bax and decreased Bcl-2 genes responsible for apoptosis. Our data suggested when AdSCs were exposed to methamphetamine, it decreased proliferation and differentiation properties of stem cells together with an increase in apoptosis. These findings can be added to the literature, especially when methamphetamine is used recreationally for weight loss purposes.

Study on the endometrium regeneration treatment using human injured uterine tissue model

The disorder of endometrial tissue regeneration is the main cause of uterine adhesions, amenorrhea, and infertility in women. Monthly changes in the endometrial cycle and injured uterine regeneration confirm the role of stem cells, hormonal regulation as well as endometrial tissue structure. In previous studies, the use of a single or combined stem cell therapy, hormone therapy, and scaffold transplantation had been increasingly emphasized. This paper presented the evaluatation of the role of the stem cell and matrix combination in the presence of estrogen hormone in endometrial regeneration on the human injured uterine tissue model. Human uterine tissue fragments were collected and mechanically damaged for in vitro modeling. The fibrin scaffolds were fabricated and combined with, respectively, human adipose tissue stem cells (hADSCs) and human bone marrow stem cells (hBMSCs) to be treated in an established injured model in conditions with or without estrogen. Histological assessments were performed to compare and confirm the regenerative effect of the treatments after 14 days of investigation. The results showed that there was no difference in the histological structure between the control treatment (no treatment) and the single treatment with stem cells. However, when stem cells were combined with fibrin matrix in the presence of estrogen, the uterine glands were formed where the layered structure of the endometrium was clearly demonstrated. In conclusion, the endometrial regenerating effect was well established by the combined treatment of these stem cells with fibrin scaffolds in the presence of estrogen. The combination of cells and scaffolds in tissue engineering therapy gradually showed the potential applications for in vitro studies, especially in the field of infertility treatment.

Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Steroidogenic Cells by NR5A1 and Implantation into Adrenal Insufficient Mice

Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells into Steroidogenic Cells by NR5A1 and Implantation into Adrenal Insufficient Mice

Development of a stem cell spheroid‐laden patch with high retention at skin wound site

Mesenchymal stem cells such as human adipose tissue‐derived stem cells (hADSCs) have been used as a representative therapeutic agent for tissue regeneration because of their high proliferation and paracrine factor‐secreting abilities. However, certain points regarding conventional ADSC delivery systems, such as low cell density, secreted cytokine levels, and cell viability, still need to be addressed for treating severe wounds. In this study, we developed a three‐dimensional (3D) cavity‐structured stem cell‐laden system for overdense delivery of cells into severe wound sites. Our system includes a hydrophobic surface and cavities that can enhance the efficiency of cell delivery to the wound site. In particular, the cavities in the system facilitate hADSC spheroid formation, increasing therapeutic growth factor expression compared with 2D cultured cells. Our hADSC spheroid‐loaded patch exhibited remarkably improved cell localization at the wound site and dramatic therapeutic efficacy compared to the conventional cell injection method. Taken together, the hADSC spheroid delivery system focused on cell delivery, and stem cell homing effect at the wound site showed a significantly enhanced wound healing effect. By overcoming the limitations of conventional cell delivery methods, our overdense cell delivery system can contribute to biomedical and clinical applications.

ASC and SVF Cells Synergistically Induce Neovascularization in Ischemic Hindlimb Following Cotransplantation.

Previously, we reported the angio-vasculogenic properties of human stromal vascular fraction (SVF) and adipose tissue-derived mesenchymal stem cells (ASCs). In this study, we investigated whether the combination of ASCs and SVF cells exhibited synergistic angiogenic properties. We conducted quantitative (q)RT-PCR, Matrigel plug, tube formation assays, and in vivo therapeutic assays using an ischemic hind limb mouse model. Immunohistochemical analysis was also conducted. qRT-PCR results revealed that FGF-2 was highly upregulated in ASCs compared with SVF, while PDGF-b and VEGF-A were highly upregulated in SVF. Conditioned medium from mixed cultures of ASCs and SVF (A+S) cells showed higher Matrigel tube formation and endothelial cell proliferation in vitro. A+S cell transplantation into ischemic mouse hind limbs strongly prevented limb loss and augmented blood perfusion compared with SVF cell transplantation. Transplanted A+S cells also showed high capillary density, cell proliferation, angiogenic cytokines, and anti-apoptotic potential in vivo compared with transplanted SVF. Our data indicate that A+S cell transplantation results in synergistic angiogenic therapeutic effects. Accordingly, A+S cell injection could be an alternative therapeutic strategy for treating ischemic diseases.

3D-Printed PCL Scaffolds Coated with Nanobioceramics Enhance Osteogenic Differentiation of Stem Cells.

With advances in bone tissue engineering, various materials and methods have been explored to find a better scaffold that can help in improving bone growth and regeneration. Three-dimensional (3D) printing by fused deposition modeling can produce customized scaffolds from biodegradable polyesters such as polycaprolactone (PCL). Although the fabricated PCL scaffolds exhibited a lack of bioactivity and poor cell attachment on their surfaces, herein, using a simple postfabrication modification method with hydroxyapatite (HA) and bioglasses (BGs), we obtained better cell proliferation and attachment. Biological behavior and osteosupportive capacity of the 3D-printed scaffolds including PCL, PCL/HA, PCL/BG, and PCL/HA/BG were evaluated in this study, while human adipose tissue-derived mesenchymal stem cells (hADSCs) were cultured on the scaffolds. The cell morphology, attachment, and proliferation were investigated using scanning electron microscopy (SEM), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, and 4′,6-diamidino-2-phenylindole (DAPI) staining. In the next step, the ability of stem cells to differentiate into osteoblasts was evaluated by measuring alkaline phosphatase (ALP) activity, calcium deposition, and bone-related gene and protein expression. In the end, the expression levels of miR-20a, miR-125a, and their target genes were also investigated as positive and negative regulators in osteogenesis pathways. The results showed that the coated scaffolds with bioceramics present a more appropriate surface for cell adhesion and proliferation, as well as efficient potential in inducing osteoconduction and osteointegration compared to PCL alone and control. The PCL/HA/BG scaffold exhibited higher in vitro cell viability and bone formation compared to the other groups, which can be due to the synergistic effect of HA and BG. On the whole, this tricomponent 3D-printing scaffold has a promising prospect for bone tissue engineering applications.

Secretome Analysis of Rabbit and Human Mesenchymal Stem and Endothelial Progenitor Cells: A Comparative Study.

Human adipose tissue-derived mesenchymal stem cells (AT-MSCs) have been studied several years for their immunomodulatory effect through the paracrine mechanism and cytokine secretion. In combination with endothelial progenitor cells (EPCs), MSCs have great therapeutical potential for the repair of endothelium and wound healing. However, little is known about the cytokine profile of rabbit AT-MSCs or even EPCs. The aim of this study was to analyze the secretomes of these rabbit stem/progenitor cells. A large-scale human cytokine array (up to 80 cytokines) was used to identify and compare cytokines secreted into conditioned media of human and rabbit AT-MSCs as well as HUVECs and rabbit EPCs. Few cytokines were highly expressed by human AT-MSCs (TIMP-2, TIMP-1), HUVECs (MCP-1, TIMP-2, GRO, Angiogenin, IL-8, TIMP-1), or by rabbit EPCs (TIMP-2). Several cytokines have moderate expression by human (MCP-1, GRO, Angiogenin, TGF-β 2, IL-8, LIF, IL-6, Osteopontin, Osteoprotegerin) and rabbit AT-MSCs (TIMP-2, TGF-β 2, LIF, Osteopontin, IL-8, IL-5, IL-3) or by HUVECs (IL-6, MIF, TGF-β 2, GCP-2, IGFBP-2, Osteoprotegerin, EGF, LIF, PDGF-BB, MCP-3, Osteopontin, Leptin, IL-5, ENA-78, TNF-β) and rabbit EPCs (TGF-β 2, Osteopontin, GRO, LIF, IL-8, IL-5, IL-3). In conclusion, the proposed method seems to be useful for the secretome analysis of rabbit stem/progenitor cells.

Improvement of an Effective Protocol for Directed Differentiation of Human Adipose Tissue-Derived Adult Mesenchymal Stem Cells to Corneal Endothelial Cells.

Corneal disease affects 12.5 million individuals worldwide, with 2 million new cases each year. The standard treatment consists of a corneal transplantation from a human donor; however, the worldwide demand significantly exceeds the available supply. Lamellar endothelial keratoplasty, the replacement of only the endothelial layer of the cornea, can partially solve the problem. Progressive efforts have succeeded in expanding hCECs; however, the ability to expand hCECs is still limited, and new sources of CECs are being sought. Crucial advances have been achieved by the directed differentiation of embryonic or induced pluripotent stem cells, but these cells have disadvantages, such as the use of oncogenes, and are still difficult to establish. We aimed to transfer such knowledge to obtain hCECs from adipose tissue-derived adult mesenchymal stem cells (ADSC) by modifying four previously published procedures. We present several protocols capable of the directed differentiation of human ADSCs to hCECs. In our hands, the protocol by Ali et al. was the best adapted to such differentiation in terms of efficiency, time, and financial cost; however, the protocol by Wagoner et al. was the best for CEC marker expression. Our results broaden the type of cells of autologous extraocular origin that could be employed in the clinical setting for corneal endothelial deficiency.

DESIGN ALGINATE BASED BLENDS FOR LIVING COMPOSITE FIBERS TO PROMOTE WOUND HEALING

The currently used approaches in the treatment of wounds and burns have been studied for many years to eliminate problems related with mechanical strength, elasticity, biocompatibility and cost. Nowadays, fabrication of composite fibers by a fiber as core and hydrogels as shell, which can be seeded by cells is rapidly increasing. In this study, it is aimed to produce a natural polymer-based dressing that can provide controlled antibiotic release to accelerate wound healing with low cost and high efficiency. The composites have been achieved by using surgical suture as a core and alginate in the shell part, which modified with starch and gelatin. Evaluating low-cost hydrogel material such as alginate, starch and gelatin in the shell layer of composite fibers by different concentrations were investigated in addition to study their swelling and drug release behaviors. The parameters for the model of an antibiotic release that can prevent common infections can be manipulated by using a biotextile-based approach to quantify the amount of antibiotics and its release to satisfy clinical requirements. Toluidine blue and Penicillin/Streptomycin were chosen as antibiotic models for drug release experiments. Moreover, human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) were applied to evaluate cell viability experiments. Results demonstrated that alginate modified starch and gelatin can be used as low-cost and promising materials for use in biomedical applications.

Cytokines Released from Human Adipose-tissue–Derived Stem Cells By b-FGF Stimulation: Effects on Angiogenesis and Lymphatic Vessels Formation By IL-8 and CXCL1

INTRODUCTION: Recently, adipose-derived stem cells (ADSCs) have been widely used in therapeutic such as wounds, ischemic limbs, and lymphedema, due to their ability to release various cytokines. In addition, ADSCs can improve the efficiency of fat transplantation. However, the molecular mechanisms in the paracrine effect, such as the types of cytokines released from ADSCs, are largely unclear. Previously, basic fibroblast growth factor (b-FGF) was reported to increase the proliferative capacity of ADSCs. In this study, we searched for cytokines released from ADSCs upon b-FGF stimulation and examined their effects on angiogenesis and lymphatic vessels formation. METHODS: In order to analyze the maximal effect of b-FGF stimulation, ADSCs obtained from a 45-year-old African-American man were cultured in either control or serum-free medium supplemented with 20, 25, and 50 μg/mL concentrations of b-FGF, respectively and 0.5 and 4 hours of stimulation time, respectively. qRT-PCR was performed on around 200 genes using TaqMan array in order to define the optimal condition for releasing cytokines with stimulation of b-FGF as stimulation condition (SC). Secondly, ADSCs were stimulated under either SC or SC without b-FGF (as a control) followed by qRT-PCR and ELISA analyses in order to select the gene and cytokines that show at least 1.5 times higher expression level than control, respectively. Based on the results from screening experiment, ADSCs derived from a 30-year-old woman (Asian), a 55-year-old man (White), and 47-year-old man (mix of African and White) were incubated under SC, and qRT-PCR was performed for the selected cytokines. Tube formation assay was performed with the 20 ng/ml concentration of selected cytokines for HUVEC (Human Umbilical Vein Endothelial Cells), TIME-GFP (Human Microvascular Endothelial Cells expressing green fluorescence protein), and HDLEC (Human Dermal Lymphatic Endothelial Cells). Finally, formed tube area (pixels)/total view area (pixels) of a still image was calculated with ImageJ software and we defined that as the tube formation rate (%). Unpaired t-test was used for statistically analysis, with significance set at P < 0.05. RESULTS: SC was determined as a combination of serum-free medium with 20 μg per mL of b-FGF for 4-hour incubation. By qRT-PCR analysis, the gene expression of CXCL1, HB-EGF, IL1B, and IL8 were upregulated and further confirmation by ELISA revealed that the secretion of CXCL1 and IL8 was promoted by b-FGF. Promotion of CXCL1 and IL8 induction by b-FGF was observed regardless of age, race, and gender by qRT-PCR analysis. Tube formation assay demonstrated that the rate of tube formation using HUVEC, TIME-GFP, and HDLEC with CXCL1 were 2.9-, 4.0-, and 1.5-fold increase compared with the control group (without CXCL1 and IL8), respectively, which were all statistically significant (P < 0.05). CONCLUSIONS: These findings suggest that b-FGF stimulation promotes the release of CXCL1 and IL8 from ADSCs, regardless of age, race, and gender in this study. These cytokines significantly promoted angiogenesis and lymphatic vessel formation and may promote various wound healing and engraftment of transplanted fat, as well as lymphatic vessel regeneration of lymphedema.

Establishment of a Pre-vascularized 3D Lung Cancer Model in Fibrin Gel-Influence of Hypoxia and Cancer-Specific Therapeutics.

Lung cancer is the most frequently diagnosed cancer worldwide and the one that causes the highest mortality. In order to understand the disease and to develop new treatments, in vitro human lung cancer model systems which imitate the physiological conditions is of high significance. In this study, a human 3D lung cancer model was established that features the organization of a tumor with focus on tumor angiogenesis. Vascular networks were formed by co-culture of human umbilical vein endothelial cells and adipose tissue-derived mesenchymal stem cells (ASC) for 14 days in fibrin. A part of the pre-vascularized fibrin gel was replaced by fibrin gel containing lung cancer cells (A549) to form tri-cultures. This 3D cancer model system was cultured under different culture conditions and its behaviour after treatment with different concentrations of tumor-specific therapeutics was evaluated. The evaluation was performed by measurement of metabolic activity, viability, quantification of two-photon laser scanning microscopy and measurement of the proangiogenic factor vascular endothelial growth factor in the supernatant. Hypoxic conditions promoted vascularization compared to normoxic cultured controls in co- and tri-cultures as shown by significantly increased vascular structures, longer structures with a higher area and volume, and secretion of vascular endothelial growth factor. Cancer cells also promoted vascularization. Treatment with 50 µM gefitinib or 50 nM paclitaxel decreased the vascularization significantly. VEGF secretion was only reduced after treatment with gefitinib, while in contrast secretion remained constant during medication with paclitaxel. The findings suggest that the herein described 3D lung cancer model provides a novel platform to investigate the angiogenic potential of cancer cells and its responses to therapeutics. Thus, it can serve as a promising approach for the development and patient-specific pre-selection of anticancer treatment.

Cytochalasin B-induced membrane vesicles from human mesenchymal stem cells overexpressing TRAIL, PTEN and IFN-β1 can kill carcinoma cancer cells

Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) are of interest as a new vector for the delivery of therapeutic agents into the tumor microenvironment. Cell-free EV-based therapy has a number of advantages over cell-based therapy, since the use of EVs allows avoiding potential undesirable transformation associated with MSCs. MSC-derived EVs can transfer natural proteins with immunomodulatory or antitumor properties. The aim of this study was to produce vesicles from mesenchymal stem cells with simultaneous overexpression of TRAIL, PTEN and IFN-β1 and analyze its antitumor and immunomodulatory properties. In this work, a stable line of human adipose tissue-derived mesenchymal stem cells (hADSCs) with simultaneous overexpression of TRAIL, PTEN and IFN-β1 was produced. To obtain this cell line hADSCs were genetically modified with a genetic multicistronic cassette encoding TRAIL, PTEN, and IFN-β1 genes separated with a self-cleaving P2A peptide nucleotide sequence. Membrane vesicles (CIMVs) were obtained from genetically modified hADSCs using cytochalasin B treatment. Antitumor and immunomodulatory properties of the CIMVs were analyzed in vitro. It was shown that CIMVs isolated from genetically modified hADSCs overexpressing TRAIL, PTEN and IFN-β1 genes are able to activate human immune cells and induce apoptosis in various types of carcinomas in vitro. Thus, the immunomodulatory and antitumor properties of CIMVs were shown. However, further studies on animal models in vivo are required.

Effect of Hypoxic Preconditioning on Angiogenesis and Senescence in Human Adipose Tissue-Derived Mesenchymal Stem Cells

Effect of Hypoxic Preconditioning on Angiogenesis and Senescence in Human Adipose Tissue-Derived Mesenchymal Stem Cells

Genetic profiling of human bone marrow and adipose tissue-derived mesenchymal stem cells reveals differences in osteogenic signaling mediated by graphene

BackgroundIn the last decade, graphene surfaces have consistently supported osteoblast development of stem cells, holding promise as a therapeutic implant for degenerative bone diseases. However, until now no study has specifically examined the genetic changes when stem cells undergo osteogenic differentiation on graphene.ResultsIn this study, we provide a detailed overview of gene expressions when human mesenchymal stem cells (MSCs) derived from either adipose tissue (AD-MSCs) or bone marrow (BM-MSCs), are cultured on graphene. Genetic expressions were measured using osteogenic RT2 profiler PCR arrays and compared either over time (7 or 21 days) or between each cell source at each time point. Genes were categorized as either transcriptional regulation, osteoblast-related, extracellular matrix, cellular adhesion, BMP and SMAD signaling, growth factors, or angiogenic factors. Results showed that both MSC sources cultured on low oxygen graphene surfaces achieved osteogenesis by 21 days and expressed specific osteoblast markers. However, each MSC source cultured on graphene did have genetically different responses. When compared between each other, we found that genes of BM-MSCs were robustly expressed, and more noticeable after 7 days of culturing, suggesting BM-MSCs initiate osteogenesis at an earlier time point than AD-MSCs on graphene. Additionally, we found upregulated angiogenic markers in both MSCs sources, suggesting graphene could simultaneously attract the ingrowth of blood vessels in vivo. Finally, we identified several novel targets, including distal-less homeobox 5 (DLX5) and phosphate-regulating endopeptidase homolog, X-linked (PHEX).ConclusionsOverall, this study shows that graphene genetically supports differentiation of both AD-MSCs and BM-MSCs but may involve different signaling mechanisms to achieve osteogenesis. Data further demonstrates the lack of aberrant signaling due to cell-graphene interaction, strengthening the application of specific form and concentration of graphene nanoparticles in bone tissue engineering.Graphic abstract

Zone-Dependent Architecture and Biochemical Composition of Decellularized Porcine Nasal Cartilage Modulate the Activity of Adipose Tissue-Derived Stem Cells in Cartilage Regeneration.

Previous anatomical studies have shown different functional zones in human nasal septal cartilage (NC). These zones differ in respect to histological architecture and biochemical composition. The aim of this study was to investigate the influence of these zones on the fate of stem cells from a regenerative perspective. Therefore, decellularized porcine septal cartilage was prepared and subjected to histological assessment to demonstrate its equivalence to human cartilage. Decellularized porcine NC (DPNC) exposed distinct surfaces depending on two different histological zones: the outer surface (OS), which is equivalent to the superficial zone, and the inner surface (IS), which is equivalent to the central zone. Human adipose tissue-derived stem cells (ASCs) were isolated from the abdominal fat tissue of five female patients and were seeded on the IS and OS of DPNC, respectively. Cell seeding efficiency (CSE), vitality, proliferation, migration, the production of sulfated glycosaminoglycans (sGAG) and chondrogenic differentiation capacity were evaluated by histological staining (DAPI, Phalloidin, Live-Dead), biochemical assays (alamarBlue®, PicoGreen®, DMMB) and the quantification of gene expression (qPCR). Results show that cell vitality and CSE were not influenced by DPNC zones. ASCs, however, showed a significantly higher proliferation and elevated expression of early chondrogenic differentiation, as well as fibrocartilage markers, on the OS. On the contrary, there was a significantly higher upregulation of hypertrophy marker MMP13 (p < 0.0001) and GAG production (p = 0.0105) on the IS, whereas cell invasion into the three-dimensional DPNC was higher in comparison to the OS. We conclude that the zonal-dependent distinct architecture and composition of NC modulates activities of ASCs seeded on DPNC. These findings might be used for engineering of cartilage substitutes needed in facial reconstructive surgery that yield an equivalent histological and functional structure, such as native NC.

Role of peroxiredoxin 6 in the chondroprotective effects of microvesicles from human adipose tissue-derived mesenchymal stem cells.

BackgroundOsteoarthritis (OA) is a joint disease characterized by cartilage degradation, low-grade synovitis and subchondral bone alterations. In the damaged joint, there is a progressive increase of oxidative stress leading to disruption of chondrocyte homeostasis. The modulation of oxidative stress could control the expression of inflammatory and catabolic mediators involved in OA. We have previously demonstrated that extracellular vesicles (EVs) present in the secretome of human mesenchymal stem cells from adipose tissue (AD-MSCs) exert anti-inflammatory and anti-catabolic effects in OA chondrocytes. In the current work, we have investigated whether AD-MSC EVs could regulate oxidative stress in OA chondrocytes as well as the possible contribution of peroxiredoxin 6 (Prdx6).MethodsMicrovesicles (MV) and exosomes (EX) were isolated from AD-MSC conditioned medium by differential centrifugation with size filtration. The size and concentration of EVs were determined by resistive pulse sensing. OA chondrocytes were isolated from knee articular cartilage of advanced OA patients. 4-Hydroxynonenal adducts, IL-6 and MMP-13 were determined by enzyme-linked immunosorbent assay. Expression of Prdx6 and autophagic markers was assessed by immunofluorescence and Western blotting. Prdx6 was downregulated in AD-MSCs by transfection with a specific siRNA.ResultsMV and to a lesser extent EX significantly reduced the production of oxidative stress in OA chondrocytes stimulated with IL-1β. Treatment with MV resulted in a dramatic upregulation of Prdx6. MV also enhanced the expression of autophagy marker LC3B. We downregulated Prdx6 in AD-MSCs by using a specific siRNA and then MV were isolated. These Prdx6-silenced MV failed to modify oxidative stress and the expression of autophagy markers. We also assessed the possible contribution of Prdx6 to the effects of MV on IL-6 and MMP-13 production. The reduction in the levels of both mediators induced by MV was partly reverted after Prdx6 silencing.ConclusionOur results indicate that EVs from AD-MSCs regulate the production of oxidative stress in OA chondrocytes during inflammation. Prdx6 may mediate the antioxidant and protective effects of MV.The translational potential of this article: This study gives insight into the protective properties of EVs from AD-MSCs in OA chondrocytes. Our findings support the development of novel therapies based on EVs to prevent or treat cartilage degradation.

Impact of human adipose tissue-derived stem cells on dermatofibrosarcoma protuberans cells in an indirect co-culture: an in vitro study

BackgroundAutologous adipose tissue transfer may be performed for aesthetic needs following the resection of dermatofibrosarcoma protuberans (DFSP), the most common cutaneous soft tissue sarcoma, excluding Kaposi sarcoma. The regenerative effectiveness of cell-assisted lipotransfer is dependent on the presence of adipose tissue-derived stem cells (ADSCs). This is the first study to evaluate the potential oncological risks as ADSCs could unintentionally be sited within the proximity of the tumor microenvironment of DFSP cells.MethodsPrimary DFSP cells were indirectly co-cultured with ADSCs in a conditioned medium or in a Transwell system. The impact was analyzed by assessing proliferation, migration, invasion, angiogenesis, and tumor-associated genes and proteins. Results of these assays were compared between co-culture and mono-culture conditions.ResultsOur experimental results showed that ADSCs were able to promote proliferation, migration, invasion, and angiogenesis of DFSP cells; this was accompanied by a significant increase in the expression levels of beta-type platelet-derived growth factor receptor, collagen type I alpha 1 chain, vascular endothelial growth factor, hepatocyte growth factor, and basic fibroblast growth factor.ConclusionsThe current report clearly demonstrates that ADSCs can enhance different malignant properties of DFSP cells in vitro, which should not be neglected when considering the clinical use of human ADSCs and its related derivatives in skin regenerative therapies.

Influence of Charge and Heat on the Mechanical Properties of Scaffolds from Ionic Complexation of Chitosan and Carboxymethyl Cellulose.

As one of the most abundant, multifunctional biological polymers, polysaccharides are considered promising materials to prepare tissue engineering scaffolds. When properly designed, wetted porous scaffolds can have biomechanics similar to living tissue and provide suitable fluid transport, both of which are key features for in vitro and in vivo tissue growth. They can further mimic the components and function of glycosaminoglycans found in the extracellular matrix of tissues. In this study, we investigate scaffolds formed by charge complexation between anionic carboxymethyl cellulose and cationic protonated chitosan under well-controlled conditions. Freeze-drying and dehydrothermal heat treatment were then used to obtain porous materials with exceptional, unprecendent mechanical properties and dimensional long-term stability in cell growth media. We investigated how complexation conditions, charge ratio, and heat treatment significantly influence the resulting fluid uptake and biomechanics. Surprisingly, materials with high compressive strength, high elastic modulus, and significant shape recovery are obtained under certain conditions. We address this mostly to a balanced charge ratio and the formation of covalent amide bonds between the polymers without the use of additional cross-linkers. The scaffolds promoted clustered cell adhesion and showed no cytotoxic effects as assessed by cell viability assay and live/dead staining with human adipose tissue-derived mesenchymal stem cells. We suggest that similar scaffolds or biomaterials comprising other polysaccharides have a large potential for cartilage tissue engineering and that elucidating the reason for the observed peculiar biomechanics can stimulate further research.

Adipose-Derived Stem Cell Features and MCF-7.

Human adipose tissue-derived stem cells (hADSCs) are highly suitable for regeneration therapies being easily collected and propagated in vitro. The effects of different external factors and culturing conditions are able to affect hADSC proliferation, senescence, differentiation, and migration, even at the molecular level. In the present paper, we exposed hADSCs to an exhausted medium from the breast cancer cell line (MCF-7) to evaluate whether the soluble factors released by these cells may be able to induce changes in stem cell behavior. In particular, we investigated the expression of stemness-related genes (OCT4; Sox 2; Nanog), the cell-cycle regulators p21 (WAF1/CIP1) p53, epigenetic markers (DNMT1 and Sirt1), and autophagy-related proteins. From our results, we can infer that the exhausted medium from MCF-7 is able to influence the hADSCs behavior increasing the expression of stemness-related genes, cell proliferation, and autophagy. Polyamines detectable in MCF-7 exhausted medium could be related to the higher proliferation capability observed in hADSCs, suggesting direct crosstalk between these molecules and the observed changes in stem cell potency.