saeed azandeh

For tissue engineering and clinical translation strategies, it is essential to have a reliable and safe lineage-specific differentiation of stem cells. To deal with several problems caused by growth factor delivery systems and growth factors, exosomes have been used as biomimetic tools to trigger the differentiation of stem cells. It is believed that cell type-specific exosomes can induce lineage-specific differentiation of stem cells. Exosomes trigger cell viability, cell proliferation and differentiation, embryonic implantation, and migration. They have been used successfully in regenerative medicine, such as liver fibrosis, renal diseases, cardiac ischemia, stroke, and skin injuries. The findings highlighted the necessity to take into account the condition and source of exosome donor cells before selecting them for therapeutic use.

Human Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs) have been recognized as a potential tool to replace damaged cells by improving the survival of the dopaminergic cells in Parkinson’s disease (PD). In this study, we examined the effects of hWJ-MSCs and associated with L-dopa/carbidopa on motor disturbances in the PD model. PD was induced by injection of 6-hydroxydopamine (6-OHDA) (16 μg/2 μl into medial forebrain bundle (MFB)). Sham group received a vehicle instead of 6-OHDA. PD+C group received hWJ-MSCs twice on the 14th and 28th days post PD induction. PD+C+D group received hWJ-MSCs and also L-dopa/carbidopa (10/30 mg/kg). PD+D group received L-dopa/carbidopa alone. Four months later, motor activities (the parameters of  locomotor and muscle stiffness) were evaluated, dopaminergic neurons were counted in substantia nigra pars compacta (SNc), the level of dopamine (DA), and tyrosine hydroxylase (TH) were measured in the striatum.   Data indicated that motor activities, the number of dopaminergic neurons, and levels of DA and TH activities were significantly reduced in PD rats as compared to the sham group (p <0.001). However, the same parameters were improved in the treated groups when compared with the PD group (p The chronic treatment of PD rats with hWJ-MSCs and L-dopa/carbidopa, improved motor activity, which may be the result of increased TH activity and due to released DA from dopaminergic neurons.

Wounds have a bad prognostic nature and excessive discharges whose regular wound dressings are ineffective. Hydrogels are the best candidates for dressing such wounds due to their high water content and ability to exchange substances. Accordingly, the purpose of this study was to make a novel hydrogel wound dressing following the integration of various findings on wound healing and the use of regenerative medicine.

Various compounds were fabricated by glycerol/chitosan/polyvinyl alcohol (PVA) and then characterized to obtain the optimal composition using several techniques, including a water vapor passage test, scanning electron microscopy, water absorption, tensile strength, biodegradability, Fourier transform infrared spectroscopy, and antibacterial test.

The findings revealed the optimal dressing ratio. Better antibacterial activity was found for the silver nanoparticle (AgNP) dressing.

Our new fabricated dressing, glycerol/chitosan/PVA hydrogel loaded with AgNPs, exhibited satisfactory wound healing properties.

There are several differentiation methods for mesenchymal stem cells (MSCs) into hepatocyte-like cell. Investigators reported various hepatic differentiation protocols such as modifying culturing conditions or using various growth factors/cytokines. In this literature review, we compared different MSCs extraction and isolation protocols from Wharton’s jelly (WJ) and explored various MSCs differentiation methods. Various protocols have been recommended for MSCs isolated from WJ, such as enzymatic, enzymatic-explant, and explant methods. In the explant method, valuable time is wasted, but the cost and biological contaminations are reduced and the number of isolated cells is high. However, other features, such as immune phenotype and multiline-age differentiation capacity, do not differ from other methods. There are also several differentiation methods for hepatocyte-like cell including the induction of MSC by cytokines and growth factors, and the differentiation of MSC in 2- and 3-dimensional matrix (2D and 3D). Among several cytokines, hepatocyte growth factor (HGF) and fibroblast growth factor (FGF) are essential. In the early stage of the differentiation, 2D culture is useful, and in the development stage, 3D culture system with HGF and FGF cytokines are more effective in the process of differentiation. Some studies have used 3D culture system in biocompatible scaffolds, such as alginate, collagen, gelatin, and peptide-Gly-Leu-amide (PGLA).In conclusion, Wharton’s jelly-Mesenchymal stem cells (WJ-MSCs) can be considered as an appropriate source for hepatocyte differentiation. Moreover, we introduced the explant method as the most effective protocol. This review attempted to highlight factors in hepatocyte differentiation, but the most effective protocol is not still unknown.

In this study, effects of encapsulated umbilical cord stem cells (UCSCs)-derived hepatocyte-like cells (HLCs) in high mannuronic alginate scaffolds was investigated on CCl4-induced acute liver failure (ALF) in rats.
Material and UCSCs were encapsulated in high mannuronic alginate scaffolds. Then the UCSCs differentiated into HLCs for treatment of CCl4-induced ALF in rats. Thirty rats randomly divided into 5 groups: Intoxicated group received only CCl4 to induce ALF. In other groups including cell-free, UCSCs and HLCs, alginate scaffolds were transplanted into the liver 4 days after CCl4 injection. Biochemical markers including albumin (ALB), blood urea nitrogen (BUN), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were evaluated. Histological changes and gene expression of ALB, alpha-fetoprotein (AFP), and cytokeratin 18 (CK-18) were also assessed.
Expression of CK-18 significantly increased in HLCs compared to the UCSCs in vitro. This indicates that UCSCs can effectively differentiate into the HLCs. In CCl4-intoxicated group, BUN, AST and ALT levels, and histological criteria, such as infiltration of inflammatory cells, accumulation of reticulocytes, nuclear pyknosis of hepatocyte and sinusoidal dilation, significantly increased. In this group, ALB secretion significantly decreased, while AFP expression significantly increased. Both UCSCs and HLCs encapsulated in alginate scaffolds effectively attenuated biochemical tests, improved liver cytoarchitecture, increased expression of ALB and reduced AFP expression.
Finding of the present study indicated that encapsulation of UCSCs or HLCs in alginate mannuronic scaffolds effectively improve CCl4-induced ALF.

Human umbilical cord Wharton’s jelly-derived mesenchymal stem cells are one of the valuable sources for cell therapy and tissue engineering, and for these purposes, it is necessary to proliferate them in culture medium. Culture medium imposes oxidative stress on cells. Vitamin C (vit C) is a potent antioxidant. The aim of this study was to investigate the effect of vit C on proliferation and differentiation of these cells.
After isolation and culture of cells from umbilical cord Wharton’s jelly, cells of third passage culture, were treated with different concentrations of vit C in five groups, including: 1- without vit C, 2- supplemented medium with 5µM of vit C; 3- supplemented medium with 50µM of vit C; 4- supplemented medium with 250, and 5- supplemented medium with 500µM of vit C. The treatment period was 9days. Cellular bioviability was assessed by MTT assay, and their differentiation potential was assessed by osteogenic and adipogenic differentiation inducer medium and histochemical staining.
In this study, vit C with concentration of 250µM increased cellular bioviability, while other concentrations decreased it (p≤0.05). Also, 50µM concentration improved osteogenic differentiation; while, in terms of adipogenic differentiation, it could just uniform dispersion of lipid droplets with 50 µM concentration.
The results of this study showed that an appropriate concentration of vit C can increase the viability of Wharton’s jelly and affect osteogenic and adipogenic differentiation in a dose-dependent manner.

The goal of the study described here, was to investigate the potential of umbilical cord derived mesenchymal stem cell (UC-MSCs) into hepatocyte like cells in a sequential 2D and 3D differentiation protocols as alternative therapy.
Mesenchymal stem cells (MSCs) were isolated from the umbilical cord (UC) and CD markers were analyzed by flow cytometry. For hepatic differentiation of UC-MSCs, cells were induced with a sequential 4-step protocol in 3D and 2D culture system. Urea concentration and albumin secretion into the culture medium was quantified by ELISA. Gene expression levels of AFP, ALB, and CK18 were determined by RT-PCR. Data were statistically analyzed by the SPSS software. The difference between the mean was considered significant when p Growth factor dependent morphological changes from elongated fibroblast-like cells to round epithelial cell morphology were observed in 2D culture. Cell proliferation analysis showed round-shaped morphology with clear cytoplasm and nucleus on the alginate scaffold in 3D culture. The mean valuses of albumin production and urea secretion were significantly higher in the 3D Culture system when compared with the 2D culture (p = 0.005 vs p = 0.001), respectively. Treatment of cells with TSA in the final step of differentiation induced an increased expression of CK18 and a decreased expression of αFP in both the 3D and 2D cultures (p = 0.026), but led to a decreased albumin gene expression, and an increased expression in the 2D culture (p = 0.001).
Findings of the present study indicated that sequential exposure of UC-MSCs with growth factors in 3D culture improves hepatic differentiation.

Umbilical cord derived mesenchymal stem cells (MSCs) are one of the best sources for cell-based therapies. MSCs reveal the neuronal morphology and expression of neuronal marker in differentiation conditions. The aim of the present study was to differentiate of MSCs into neuron-like cells using Activin A and nicotinamide.
Umbilical cords were cut close to the placenta and smaller pieces (2-4 cm) of umbilical cords prepared and Wharton's jelly extracted. MSCs were isolated by an explant culture method and surface phenotype of cells analyzed by Dako flow cytometry and the FlowJo software. MSCs were then induced for osteogenic and adipogenic differentiation. Cells were exposed to neuronal differentiation condition and treated with 20 μg / ml Activin A and 0.1% FBS for 3 days in RPMI,then with 10 mMnicotinamide, 2% B27 and 0.1% FBS for 7 days.
spindle-shaped MSCs were observed after 7-10 days of explants culture. Immunophenotype of cultured MSCs were positive for mesenchymal markers such as CD105 and CD90 but negative for hematopoietic markers such as CD34 and CD45. After MSCs differentiation to the osteogenic or adipogenic lineage, lipid droplets and calcium deposition were observed. The differentiated neural-like MSCs were appeared as pyramidal nerve-like cells with neuritis.
Some of th cells. During neuronal differentiation, axon and dendritic like process wereosbserved in some of Bipolar and mulipolarmulipolar cells. Furthermore direct synaptic like contacts were observed between cells.
The present study showed that Activin A and nicotinamide could induce differentiation of MSCs into neural lineage.

Due to increasing demand for liver tissue engineering, three-dimensional (3D) liver cells culture techniques have been proposed. Therefore, the aim of the present study was to examine the cells isolation and expansion of umbilical cord derived mesenchymal stem cells and in vitro 2D and 3D hepatocyte differentiation. Also functional characteristics of hepatocytes were analyzed. The study performed in several phases. In the first umbilical cord derived mesenchymal stem cells obtained and isolated, thereafter cellss expanded. Determination of Immunophenotype using Flowcytometry performed by DAKO – Galaxy Hepatic differentiation UC-MSCs was performed by four step sequential method using FGF-4, ITS, HGF, dexamethasone, glucagon, OSM and TSA. Urea production was quantified by ELISA. Section of tissue constructs stained with hematoxyllin and eosin for histological examination. MSCs isolated from umbilical cord expressed mesenchymal surface antigen such as CD73, but were negative against CD31. Several cell clusters mainly between the round cells were observed in alginate scaffold after 3d differentiation. Urea production was increased time- dependable and was significantly higher in the experimental group of 3D culture (P=0.001). Tissue construct of 3D culture revealed multicellular tissue with several euchromatin cell plates. The finding of the present study indicated that four step differentiation of umbilical cord derived mesenchymal stem cells within hydrogel scaffold induced functionally and morphologically characteristics of hepatocytes such as urea production and cell plates.