Induction of spontaneous neo-angiogenesis and tube formation in human endometrial stem cells by bioglass

Abstract Endothelial dysfunction is a broad pathological disorder of the endothelium (innermost layer of blood vessels) which is assigned by vasoconstriction, thrombosis and ischemic diseases, alone or with other disorders such as coronary artery disease, hypertension and atherosclerosis. The fundamental imperfection of endothelial layer injury due to decrease in the number of functional endothelial progenitor cells and inhibition of endothelial progenitor cell differentiation, resulting into impairment of angiogenesis, vasculogenesis, tube formation properties and endothelial regeneration. Multiple significant therapeutic achievements in impediment and treatment of vascular diseases include the use of antithrombotic agents, statin class of drugs, lifestyle changes, and revascularization therapies. Nevertheless, a certain number of patients with endothelial dysfunction disease are resistant to the usual therapies, so new therapeutic strategies for endothelial dysfunction disease are urgently needed. Recent studies show that stem cell-based therapy has important promise for repair and treatment of vascular dysfunction. In this study, we describe a novel choice for treatment of endothelial dysfunction in vascular regenerative medicine via the human endometrial stem cell culture (as a new source for the increasing the number of endothelial progenitor cells) with bioglass (angiogenic agent) to investigate the enhancing expression of CD34, CD31 and gene markers of endothelial progenitor cells and endothelial cells. In the end, application of immunoprivileged, readily available sources of adult stem cells like human endometrial stem cells with bioglass would be a promising strategy to increase the number of endothelial progenitor cells and promote spontaneous angiogenesis needed in endothelial layer repair and regeneration. �2015 Tehran University of Medical Sciences. Published by Elsevier Ltd. This is an open access articl

Lentiviral Mediated Overexpression of NGF in Adipose-derived Stem Cells

Introduction: Human adipose-derived stem cells (ADSCs) are multipotent stem cells that can self-renew and differentiate into various types of cells such as adipocytes, osteocytes, and neural cells. These stem cells can be isolated by minimally invasive technique in large amounts. ADSCs are a useful resource for cell therapy and regenerative medicine. Nerve growth factor (NGF) is the first neurotrophin factor discovered and characterized for its anti-apoptotic role in neural development. NGF can promote neuronal survival and neurite outgrowth and it also promotes neuron differentiation and migration. Moreover, research showed that NGF could protect axons from inflammatory damage, improve cognitive function in damaged brain models, and function in the prevention and treatment of neurological diseases like Alzheimer’s disease. In this study we use Lentiviral vector-mediated gene transfer technique to deliver NGF gene to ADSCs and overexpress this factor in ADSCs. Method and Materials: ADSCs extracted from human adipose tissue after lipoaspiration by digestion method. ADSCs characterized with Flowcytometry and differentiation assay in adipogenic and osteogenic differential media. The NGF gene was cloned in pCDH-513B-1 (System Bioscience, Mountain View, CA, United States) under a cytomegalovirus (CMV) promoter. Recombinant lentiviruses were produced according to the Prof. Trono lab protocol with some modifications in HEK 293T cells. The spinfection method was used to transduce ADSCs. NGF expression was assayed using fluorescent microscope to trace green fluorescent protein (GFP) marker, RT-PCR and western blotting. Results: Extracted ADSCs had mesenchymal morphology and differentiated into adipocytes and osteocytes in differentiating media. HEK293T easily transfected with pCDH-513B-1 and over 99% of them expressed GFP so we gathered pseudoviruses from the supernatant. ADSCs transduced with these pseudoviruses transferred NGF and after transduction expressed GFP, as seen under fluorescent microscope. RT-PCR and western blotting verified NGF overexpression in them

In vitro evaluation of human endometrial stem cell-derived osteoblast-like cells’ behavior on gelatin/collagen/bioglass nanofibers’ scaffolds

New biomimetic nanocomposite scaffold was prepared by the combination of nanofibrilar bioglass containing copper ion as the inorganic phase and gelatin/collagen as the organic phase of bone tissue. In this study for fabrication of the scaffold, freeze drying and electrospinning methods were used, and genipin was used as the cross-linking agent for increasing the mechanical properties of the scaffold. The growth and viability of human endometrial stem cell-derived osteoblast-like cells were investigated on this biomimetic scaffold. Cellular biocompatibility assays illustrated that this scaffold has more viabilities and osteoblast growths in comparison with two-dimensional culture. Copper ion increased growth of the osteoblasts on nanocomposite scaffold containing nanofibrous bioglass. Thus, the results obtained from this study indicate that the prepared scaffold is suitable for osteoblast growth and attachment; thus, potentially, this nanocomposite scaffold is an appropriate scaffold for bone tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2210–2219, 2016

Nanotoxicology and nanoparticle safety in biomedical designs

Nanotechnology has wide applications in many fields, especially in the biological sciences and medicine. Nanomaterials are applied as coating materials or in treatment and diagnosis. Nanoparticles such as titania, zirconia, silver, diamonds, iron oxides, carbon nanotubes, and biodegradable polymers have been studied in diagnosis and treatment. Many of these nanoparticles may have toxic effects on cells. Many factors such as size, inherent properties, and surface chemistry may cause nanoparticle toxicity. There are methods for improving the performance and reducing toxicity of nanoparticles in medical design, such as biocompatible coating materials or biodegradable/biocompatible nanoparticles. Most metal oxide nanoparticles show toxic effects, but no toxic effects have been observed with biocompatible coatings. Biodegradable nanoparticles are also used in the efficient design of medical materials, which will be reviewed in this article

Mesenchymal stromal cells induce inhibitory effects on hepatocellular carcinoma through various signaling pathways

Hepatocellular carcinoma (HCC) is the most prevalent type of malignant liver disease worldwide. Molecular changes in HCC collectively contribute to Wnt/β-catenin, as a tumor proliferative signaling pathway, toll-like receptors (TLRs), nuclear factor-kappa B (NF-κB), as well as the c-Jun NH2-terminal kinase (JNK), predominant signaling pathways linked to the release of tumor-promoting cytokines. It should also be noted that the Hippo signaling pathway plays an important role in organ size control, particularly in promoting tumorigenesis and HCC development. Nowadays, mesenchymal stromal cells (MSCs)-based therapies have been the subject of in vitro, in vivo, and clinical studies for liver such as cirrhosis, liver failure, and HCC. At present, despite the importance of basic molecular pathways of malignancies, limited information has been obtained on this background. Therefore, it can be difcult to determine the true concept of interactions between MSCs and tumor cells. What is known, these cells could migrate toward tumor sites so apply efects via paracrine interaction on HCC cells. For example, one of the inhibitory efects of MSCs is the overexpression of dickkopf-related protein 1 (DKK-1) as an important antagonist of the Wnt signaling pathway. A growing body of research challenging the therapeutic roles of MSCs through the secretion of various trophic factors in HCC. This review illustrates the complex behavior of MSCs and precisely how their inhibitory signals interface with HCC tumor cells. Keywords: Mesenchymal stromal cells, Hepatocellular carcinoma, Wnt signaling, Toll like receptor, Nuclear factorkappa B, JNK pathwa

Differentiation of human endometrial stem cells encapsulated in alginate hydrogel into oocyte-like cells

Introduction: Human endometrial mesenchymal stem cells (hEnMSCs) are a rich source of mesenchymal stem cells (MSCs) with multi-lineage differentiation potential, making them an intriguing tool in regenerative medicine, particularly for the treatment of reproductive and infertility issues. The specific process of germline cell-derived stem cell differentiation remains unknown, the aim is to study novel ways to achieve an effective differentiation method that produces adequate and functioning human gamete cells. Methods: We adjusted the optimum retinoic acid (RA) concentration for enhancement of germ cell-derived hEnSCs generation in 2D cell culture after 7 days in this study. Subsequently, we developed a suitable oocyte-like cell induction media including RA and bone morphogenetic protein 4 (BMP4), and studied their effects on oocyte-like cell differentiation in 2D and 3D cell culture media utilizing cells encapsulated in alginate hydrogel. Results: Our results from microscopy analysis, real-time PCR, and immunofluorescence tests revealed that 10 µM RA concentration was the optimal dose for inducing germ-like cells after 7 days. We examined the alginate hydrogel structural characteristics and integrity by rheology analysis and SEM microscope. We also demonstrated encapsulated cell viability and adhesion in the manufactured hydrogel. We propose that in 3D cell cultures in alginate hydrogel, an induction medium containing 10 µM RA and 50 ng/mL BMP4 can enhance hEnSC differentiation into oocyte-like cells. Conclusion: The production of oocyte-like cells using 3D alginate hydrogel may be viable in vitro approach for replacing gonad tissues and cells

Li-Doped Bioactive Ceramics:Promising Biomaterials for Tissue Engineering and Regenerative Medicine

Lithium (Li) is a metal with critical therapeutic properties ranging from the treatment of bipolar depression to antibacterial, anticancer, antiviral and pro-regenerative effects. This element can be incorporated into the structure of various biomaterials through the inclusion of Li chloride/carbonate into polymeric matrices or being doped in bioceramics. The biocompatibility and multifunctionality of Li-doped bioceramics present many opportunities for biomedical researchers and clinicians. Li-doped bioceramics (capable of immunomodulation) have been used extensively for bone and tooth regeneration, and they have great potential for cartilage/nerve regeneration, osteochondral repair, and wound healing. The synergistic effect of Li in combination with other anticancer drugs as well as the anticancer properties of Li underline the rationale that bioceramics doped with Li may be impactful in cancer treatments. The role of Li in autophagy may explain its impact in regenerative, antiviral, and anticancer research. The combination of Li-doped bioceramics with polymers can provide new biomaterials with suitable flexibility, especially as bio-ink used in 3D printing for clinical applications of tissue engineering. Such Li-doped biomaterials have significant clinical potential in the foreseeable future