somaieh kazemnejad

In this study we differentially showed the effects of cell-seeded bilayer scaffold wound dressing in accelerating healing process in diabetic ulcers that still remains as a major clinical challenge. The aim of the study was to compare immunomodulatory and angiogenic activity, and regenerative effect differences between Menstrual blood-derived Stem Cells (MenSCs) and foreskin-derived keratinocytes/fibroblasts.

The streptozotocin-induced diabetic mice model was developed in male C57/BL6 mice. A bilayer scaffold was fabricated by electrospining silk fibroin nanofibers on human Amniotic Membrane (AM). Dermal fibroblasts and keratinocyte isolated from neonatal foreskin and MenSCs were isolated from the menstrual blood of healthy women. The diabetic mice were randomly divided into three groups including no treatment group, fibroblast/keratinocyte-seeded bilayer scaffold group (bSC+FK), and MenSCs-seeded bilayer scaffold group. The healing of full-thickness excisional wounds evaluations in the diabetic mice model in each group were evaluated at 3, 7, and 14 days after treatment.

The gross and histological data sets significantly showed wound healing promotion via re-epithelialization and wound contraction along with enhanced regeneration in MenSCs-seeded bilayer scaffold group with the most similarity to adjacent intact tissue. Immunofluorescence staining of mouse skin depicted a descending trend of type III collagen along with the higher expression of involucrin as keratinocyte marker in the MenSCs-seeded bilayer nanofibrous scaffold group in comparison with other treatment groups from day 7 to day 14. Moreover, higher levels of CD31 and von Willebrand factor (VWF), and also a higher ratio of M2/M1 macrophages in association with higher levels of the neural marker were observed in the bSC+MenSCs group in comparison with bSC+FK and no treatment groups.

Healing symptoms in wounds dressed with keratinocyte/fibroblast-seeded bilayer scaffold was significantly lower than MenSCs-seeded bilayer scaffold done on impaired diabetic wound chronicity.

To evaluate the efficiency of Menstrual blood Stromal/Stem Cells (MenSCs) administration in Myocardial Infarction (MI), the effects of MenSCs and their derived conditioned Medium (CM) on cardiac function in MI rat model was assessed.

Animals were divided into four groups including sham group, MI group, MenSCs derived CM group (CM group), and MenSCs suspended in CM (MenSCs+ CM) group. The injection of different groups was carried out 30 min after ligation of left anterior descending coronary artery into the infarct border zone.

The results showed a significant reduction in scar size after injection of MenSCs+CM compared to MI group. Ejection fraction and fractional shortening of MenSCs+CM group were higher than CM and MI group at day 28. Administration of MenSCs+CM led to much more survival of cardiomyocytes, and prevention of metaplastic development. Moreover, human mitochondrial transfer from MenSCs to cardiomyocytes was seen in group treated by MenSCs+CM. Indeed, MenSCs+CM treatment evoked nuclear factor-κB (NF-κB) down-regulation more than other treatments.

MenSCs+CM treatment could significantly ameliorate cardiac function by different mechanisms including inhibition of cartilaginous metaplasia, inhibition of NF- κB and mitochondrial transfer.

Remote organ injury is a phenomenon that could happen following myocardial infarction (MI). We evaluated the potency of menstrual blood stromal (stem) cells (MenSCs) and bone marrow stem cells (BMSCs) to alleviate remote organ injuries following MI in rats.

2 × 106 MenSCs or BMSCs were administrated seven days after MI induction via the tail vein. Four weeks after cell therapy, activities of aspartate aminotransferase (AST), urea, creatinine, and Blood Urea Nitrogen (BUN) were evaluated. The level of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 were determined by ELISA assay. The expression of Nuclear Factor-κB (NF-κB) was evaluated by immunohistochemical staining. Apoptosis activity and tissue damage were also determined by TUNEL and H&E staining, respectively.

MenSCs and BMSCs administration caused a significant reduction in AST, urea, and BUN levels compared with the MI group. In addition, systemic injection of MenSCs significantly decreased the IL-1β level compared with BMSCs and MI groups (P<0.05 and P<0.01 respectively). Apoptosis in injured kidneys was noticeably diminished in MenSCs-treated rats compared with BMSCs administrated and MI groups (P<0.05 and P<0.05, respectively). In hepatic tissue, limited numbers of TUNEL-positive cells were detected in all groups. Interestingly, MenSCs therapy evoked inhibition of NF-κB in the kidney strikingly. Although, no significant NF-κB expression was observed in hepatic tissue in any group (P>0.05).

MenSCs are probably more protective than BMSCs on remote organ injuries following MI via decreasing cell death and immunoregulatory properties.

Premature luteinizing hormone (LH) surge is one of the causes for assisted reproductive technology cycle cancellation, and it is needed to find novel approaches with improved efficacy and safety profile.

To compare the effects of Duphaston and Cetrotide on the prevention of premature LH surge and characteristics of retrieved follicles and embryos in women undergoing intracytoplasmic sperm injection.

In this retrospective cross-sectional study, 200 patients who were administrated recombinant follicle-stimulating hormone from the third day of menstruation cycle were included. When the follicular diameter reached above 13-14 mm, Cetrotide was prescribed in the control group, while in the case group, Duphaston was taken orally from the third day of cycle. The retrieved oocytes were fertilized in vitro by intracytoplasmic sperm. The level of hormones on the third day of menstruation and the characteristic of follicles, oocytes, and embryos were compared between the two groups.

Duphaston successfully inhibits premature LH surge. There was no significant difference in the level of follicle-stimulating hormone, estradiol, and LH between the case and control groups (p > 0.05). However, results also showed that Duphaston causes more oocyte retrieval in comparison with Cetrotide (p = 0.04). Although, the number of follicles above 14 mm, mature oocyte, and the total number of viable embryos in the case group was slightly higher, it did not reach a significant difference compared with the control group (p > 0.05).

Duphaston could be used as an appropriate medication instead of gonadotropin-releasing hormone antagonists in women undergoing controlled ovarian hyperstimulation. Duphaston prescription not only prevents premature LH surge but also improves the number of retrieved oocytes.

Tissue engineering and regenerative medicine have typically matured from benchtop ideas to commercially applicable products in the clinic 1. However, despite of typical advances in tissue engineering field, some limitations such as no reproducibility, no control of structure geometry including pore size and pore distribution and no integrity of cell distribution and migration in the construct have impelled the scientists into bioprinting technology. The most advantage of 3D bioprinting sounds to be precise fabrication of 3D deposition with controlled geometric structure and cells distribution 2. Over the past decade, lots of researches in bioprinting of different tissues and organs has been carried out using different bioprinting modalities particularly inkjet based printing for skin tissue engineering and extrusion based printing for 3D depositions like bone, cartilage, heart, liver and heart valve. The key factor in extrusion-based bioprinting is bioink preparation, cell encapsulation in the bioink and bioprinting procedure. Indeed, preparation of bioink with appropriate gelation rate, suitable mechanical strength and elasticity which preserve cell viability and proliferation is the most challenge of bioprinting technology. So far, different strategies such as dual bioink cross-linkers, multi-step polymerization and using of core-shell nozzle have been reported to improve viability, quality and functionality of the printed product 3. However, some issues including creation of constructs supporting in vivo vascularization, scaling up tissue constructs and in situ bioprinting have been remained to resolve. A few bioprinting products have been commercialized especially in orthopedic and skin tissue engineering fields and given the fast development of this industry over the past years; it supposed that the bioprinting products will eventually take a big proportion of the medical market to help patients suffering from a wide range of diseases in the future.

It is more than sixty years that the concept of the fetal allograft and immunological paradox of pregnancy was proposed and in this context, several regulatory networks and mechanisms have been introduced so far. It is now generally recognized that mesenchymal stem cells exert potent immunoregulatory activity. In this study, for the first time, the potential impact of Menstrual blood Stem Cells (MenSCs), as surrogate for endometrial stem cells, on proliferative capacity of CD4 T cells was tested.
MenSCs and Bone marrow Mesenchymal Stem Cells (BMSCs) were isolated and assessed for their immunophenotypic features and multi-lineage differentiation capability. BMSCs and MenSCs with or without IFNγ pre-stimulation were co-cultured with purified anti-CD3/CD28-activated CD4 T cells and the extent of T cell proliferation at different MenSCs: T cell ratios were investigated by CSFE flow cytometry. IDO activity of both cell types was measured after stimulation with IFNγ by a colorimetric assay.
MenSCs exhibited dual mesenchymal and embryonic markers and multi-lineage differentiation capacity. MenSCs significantly increased proliferation of CD4 cells at ratios 1:2, 1:4 and 1:8. IFNγ pre-treated BMSCs but not MenSCs significantly suppressed CD4 T cells proliferation. Such proliferation promoting capacity of MenSCs was not correlated with IDO activity as these cells showed the high IDO activity following IFNγ treatment.
Although augmentation of T cell proliferation by MenSCs can be a basis for maintenance of endometrial homeostasis to cope with ascending infections, this may not fulfill the requirement for immunological tolerance to a semi-allogeneic fetus. However, more investigation is needed to examine whether or not the immunomodulatory properties of these cells are affected by endometrial microenvironment during pregnancy.

Cartilage is an avascular, aneural, and alymphatic connective tissue with a limited capacity caused by low mitotic activity of its resident cells, chondrocytes. Natural repair of full thickness cartilage defects usually leads to the formation of fibrocartilage with lower function and mechanical force compared with the original hyaline cartilage and further deterioration can occur. Tissue engineering and regenerative medicine is a promising strategy to repair bone and articular cartilage defects and rehabilitate joint functions by focusing on the optimal combination of cells, material scaffolds, and signaling molecules. The unique physical and topographical properties of nanofibrous structures allow them to mimic the extracellular matrix of native cartilage, making an appropriate resemblance to induce cartilage tissue regeneration and reconstruction. To improve simulation of native cartilage, the incorporation of nanofibrous scaffolds with suitable corresponsive cells could be effective. In this review article, an attempt was made to present the current state of cartilage tissue engineering using nanofibrous scaffolds and stem cells as high proliferative immune privilege cells with chondrogenic differentiation ability. The comprehensive information was retrieved by search of relevant subject headings in Medline/Pubmed and Elsevier databases.

It has been reported that secreted frizzled-related protein-4 known as an antagonist of Wnt signaling pathway plays a role in luteinization process of rodent granulosa cells. The purpose of this study was twofold: 1) to determine whether recombinant human secreted frizzled-related protein-4 (rhSFRP-4) could directly induce terminal differentiation of rat Granulosa Cells (GCs) and 2) to understand how the modulation of β-catenin and Protein Kinase B (PKB)/AKT activity by exogenous SFRP-4 could be involved in steroidogenesis.
GCs were firstly stimulated with Follicle-Stimulating Hormone (FSH) named as FSH-primed cells then were treated with luteinizing hormone (LH). Then estradiol (E2) and progesterone (P4) production levels were assessed in the absence or presence of rhSFRP-4 treatment. The expression levels of activated -catenin, pAKTser473, pGSK3ser9 were assessed by western blot or immuno-fluoresence.
In the presence of rhSFRP-4, there was 38% decreased E2 levels compared to untreated FSH-primed cells (p Taken together, our results showed that rhSFRP-4 could directly induce terminal differentiation in GCs via the modulation of β-catenin and PKB/AKT pathways and that it does so in a dose-dependent manner.

At present, Iran has been known one of the up-warding countries in the world in regenerative medicine using stem cells therapy. In fact, the outcomes of some clinical trials on stem cell therapy of myocardial infarction, vitiligo, decompensated cirrhosis, and osteoarthritis narrate the feasibility of stem cell-based therapy for treatment of human diseases 1,2. However, in a similar manner with global configuration, the commercialization and translation of tissue engineering products into clinical phase has been restricted. It might be due to weak collaboration of different specialties for technology transfer of the multidisciplinary projects of tissue engineering field into clinical phase. Basic tissue engineers mostly prefer elegant studies, whereas physicians have tendency to solve medical problems with products indicating efficiency, easy to use, and cost benefit. Actually, a surgeon encountered with a dilemma between a partially effective tissue-engineered product that is both expensive and difficult to apply and a more traditional approach may choose the latter option. Therefore, a coherent teamwork between basic sciences and medicine as well as acquisition of competent knowledge about target tissue is necessary to conduct tissue engineering in the clinic. Moreover, it should be considered that in developing countries including Iran the high cost of high-tech biomedical research necessitates government investment 3. Currently, the policy makers have established some action plans to support of science-based companies financially. This is a suitable opportunity to ligature basic research and market for commercialization of tissue engineering products. However, because private investors beyond academic laboratories should provide financing of tissue engineering products, incentive of private companies for investment should not be neglected. It is notable that tissue-engineered products will fulfill small market size unless they could indicate much superior results than competitive alternatives.
It is noticeable that tissue engineers should determine the requirements of community and develop strategies to penetrate the products into clinic. Indeed, the communication between scientists and policy makers should be increased to better definition of national research priorities. On the other hand, considering local necessities and natural resources should be rather than subjective experts’ notions or international superiorities. Finally, ethical and legal regulations should be actually defined that indubitably make great profits to the society.

In recent decades, stem cell therapy has been introduced as a novel therapeutic approach for patients suffering from cardiac disorders. Recently, identification of menstrual blood-derived stem cells (MenSCs) as a unique source of stem cell with some characteristics as well as ease of access, high proliferative ability and renewability has created enormous promise for cell therapy.
In this study, differentiation ability of MenSCs into cardiomyocytes has been investigated. After MenSCs immunophenotyping, their differentiation into cardiomyocyte was investigated in the presence of 5-azacytidine and basic-fibroblast growth factor. Then, expression of the putative myogenic cells at mRNA and protein levels was determined by immunofluorescent staining and real-time quantitative PCR.
Based on flow cytometric analysis, the isolated MenSCs typically expressed mesenchymal stem cell markers like CD105, CD73, CD44 and CD166 in parallel to OCT-4 as an embryonic marker. The differentiated MenSCs expressed cardiomyocyte markers at mRNA/protein level. The myogenic cells differentiated from MenSCs were positive for Connexin-43 and troponin T2 (TNNT2) protein. The mRNAs of Connexin-43, Connexin-40, Alpha actinin, Tropomyosin1 and TNNT2 were highly expressed in the differentiated myogenic cells.
Based on our data, MenSCs are a unique cell population with differentiation ability into cells with characteristics commonly attributed to cardiomyocytes.

Menstrual blood-derived stem cells (MenSCs) are a novel source of stem cells that can be easily isolated non-invasively from female volunteered donor without ethical consideration. These mesenchymal-like stem cells have high rate of proliferation and possess multi lineage differentiation potency. This study was undertaken to isolate the MenSCs and assess their potential in differentiation into epidermal lineage. About 5-10 ml of menstrual blood (MB) was collected using sterile Diva cups inserted into vagina during menstruation from volunteered healthy fertile women aged between 22-30 years. MB was transferred into Falcon tubes containing phosphate buffered saline (PBS) without Ca2+ or Mg2+ supplemented with 2.5 µg/ml fungizone, 100 µg/mL streptomycin, 100 U/mL penicillin and 0.5 mM EDTA. Mononuclear cells were separated using Ficoll-Hypaque density gradient centrifugation and washed out in PBS. The cell pellet was suspended in DMEM-F12 medium supplemented with 10% FBS and cultured in tissue culture plates. The isolated cells were co-cultured with keratinocytes derived from the foreskin of healthy newborn male aged 2-10 months who was a candidate for circumcision for differentiation into epidermal lineage. The isolated MenSCs were adhered to the plate and exhibited spindle-shaped morphology. Flow cytometric analysis revealed the expression of mesenchymal markers of CD10, CD29, CD73, and CD105 and lack of hematopoietic stem cells markers. An early success in derivation of epidermal lineage from MenSCs was visible. The MenSCs are a real source to design differentiation to epidermal cells that can be used non-invasively in various dermatological lesions and diseases.

Toll-like receptor (TLR)-mediated inflammatory processes are supposed to be involved in pathophysiology of spontaneous abortion and preterm labor. Here, we investigated functional responses of human endometrial stromal cells (ESCs) and whole endometrial cells (WECs) to lipopolysaccharide (LPS) and lipoteichoic acid (LTA). Endometrial tissues were obtained from 15 cycling women who underwent laparoscopic tubal ligation. Modulation of TLR2, TLR4 and MyD88 expression and pro-duction of pro-inflammatory cytokines by WECs and ESCs in response to LPS and LTA were assessed. WECs and ESCs expressed significant levels of TLR4 and MyD88 transcripts but, unlike WECs, ESCs failed to express TLR2 gene. Regardless of positive results of Western blotting, ESCs did not express TLR4 at their surface as judged by flow cytometry. Immunofluorescent staining revealed intracellular localization of TLR4 with predominant perinuclear pattern. LPS stimulation marginally increased TLR4 gene ex-pression in both cell types, whereas such treatment significantly upregulated MyD88 gene expression after 8 hr (p<0.05). At the protein level, however, LPS activation significantly increased TLR4 expression by ESCs (p<0.05). LTA stimulation of WECs was accompanied with non-significant increase of TLR2 and MyD88 transcripts. LPS and LTA stimulation of WECs caused significant production of IL-6 and IL-8 in a dose-dependent manner (p<0.05). Similarly, ESCs produced significant amounts of IL-6, IL-8 and also TNF-α in response to LPS activation (p<0.05). Our results provided further evidence of initiation of inflammatory processes following endometrial TLR activation by bacterial components which could potentially be harmful to developing fetus.

Despite the extensive information available in the literature, cell surface marker signature of human Amniotic Epithelial Cells (hAECs) remains controversial. The aim of the present study was to characterize immuno-phenotypic features, proliferative capacity and immunogenicity of hAECs. We also tested whether expression of some cell surface markers is influenced by the type of trypsin used for tissue digestion. Single cell suspensions of amniotic membranes from four human placentas were isolated by enzymatic digestion and expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, HLA-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. The differential impact of four trypsin types on the yield and expression of CD105 and HLA-I was also determined. The proliferative capacity of cultured hAECs was assessed and compared in the presence and absence of Epidermal Growth Factor (EGF). To test their immunogenicity, hAECs were injected into Balb/c mice and the reactivity of hyperimmunized sera was examined by immunofluorescence staining. Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73 and the embryonic marker, SSEA-4. A large proportion of the cells also expressed STRO-1 and OCT-4. The purified cells also expressed HLA-G and HLA-I. A very small proportion of hAECs expressed CD34, CD38, CD44, CD133 and HLA-DR. The type of trypsin used for enzymatic digestion affected both the percentage and expression of HLA-I and CD105. hAECs revealed substantial proliferative capacity only when cultured in the medium supplemented with EGF. These cells were shown to be capable of inducing high amounts of anti-donor antibodies. Here we provided evidence that hAECs are immunogenic cells with high level of HLA-I expression. Furthermore, this work highlighted the impact of isolation procedure on the immunophenotype of hAEC.

Currently several studies are being carried out on various properties of mesenchymal stem cells (MSCs) however there are a few investigations about drug metabolizing properties of these cells. The aim of this study was to measure the key factors involved in drug metabolism in human bone marrow MSCs. For this purpose, cellular glutathione (GSH), glutathione S-transferase (GSTs) and cytochrome P450 class 3A4 (CYP3A4) were detected in these cells. Results showed that CYP3A4 and GSTA1-1 mRNA are not detectable in MSCs however mRNA specific for GSTP1-1 was considerably expressed in MSCs. GSH content and GST activity was also detected in MSCs. These data suggest that human bone marrow MSCs possess the drug metabolizing activity which may be useful in handling drugs and chemotherapeutic agents passing to the bone marrow.

Human bone marrow derived mesenchymal stem cells (HBMSCs) have the potential to differentiate into cells such as adipocyte, osteocyte, hepatocyte and endothelial cells. In this study, the differentiation of hBMSCs into endothelial like-cells was induced in presence of vascular endothelial growth factor (VEGF) and insulin-like growth factor (IGF-1). The differentiated endothelial cells were examined for their ability to express VEGF receptor-2 (VEGFR2) and von willebrand factor (vWF). Then the cells were adopted to grow and develop capillary network in a semisolid gel matrix in vitro. The capillary network formation in a well of 24-well plate was found to be 85% in presence of VEGF (50ng/ml) and IGF-1 (20ng/ml) of the culture media. These data may suggest that the expression of endothelial markers in endothelial like-cells derived from hBMSCs is associated with their ability to form capillaries.

In this present study, we examined the differentiation potential of human bone marrow derived mesenchymal stem cells (hBMSCs) into hepatocytes on a three-dimentional (3D) nanofibrous scaffold formed by Poly (e-caprolactone) (PCL), collagen and polyethersulfone (PES). The nanofiber was prepared by the electrospining technique. HBMSCs were isolated using combining gradient density centrifugation with plastic adherence. Flow cytometric analysis was used to identify the isolated MSCs. The performance of the cells on the scaffold was evaluated by scanning electron microscopy (SEM) and MTT assay. The hBMSCs were then cultured in a hepatic differentiation medium containing hepatocyte growth factor (HGF), oncostatin M (OSM) and dexamethasone (DEX) for up to 21 days. The results showed that the isolated hBMSCs expressed specific markers such as CD44, CD166, CD105 and CD13. The integrity of the MSCs was further confirmed by their differentiation potential to osteogenic and adipogenic lineages. Scanning electron micrographs and MTT analysis revealed that the cells adhered and proliferated well on the nanofibrous hybrid scaffolds. Immunocytochemical analysis of albumin and a-fetoprotein (AFP) showed the accumulation of these markers in the differentiated cells on the scaffold. Hepatocyte differentiation was further confirmed by showing expression of albumin, AFP and cytokeratin-19 (CK-19) at mRNA levels in differentiated cells. In conclusion, the evidences presented in this study show that the engineered scaffold is promising for maintenance of hepatocyte-like cells suitable for transplantation.

Class-Pi of glutathione s-transferases (GST-Pi) is the specific form of GSTs that are known to participate particularly in the mechanisms of resistance to drugs and carcinogens.This class of the enzyme is referred to as class-P or class-Pior class? . The accepted terminology in this review article is class-Pi. In this article following a brief description of identified molecular forms of GSTs, we focus on GST-Pi. We review new findings about the structure and regulation of GST-Pi gene. Then, the role of GST-Pi in liver damage, oxidative stress, carcinogenesis and drug resistance are discussed.Also, the presence of common genetic polymorphism,hypermethylation in GST-Pi gene and the consequences GST-Pi knock out is regarded.