Medical Journal of The Islamic Republic of Iran (MJIRI)

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Ex vivo expansion of human umbilical cord blood cells (HUCBC) is explored by several investigators to enhance the repopulating potential of HUCBC. The proliferation and expansion of human hematopoietic stem cells (HSC) in ex vivo culture was examined with the goal of generating a suitable clinical protocol for expanding HSC for patient transplantation. Using primary human mesenchymal stem cells, we established a serum-free culture system to expand human primitive progenitors and transplantable stem cells. Non-enriched cord blood CD34+ cells were cultured on a monolayer of human mesenchymal stem cells in the presence of tlu-ombopoietin (TPO), flt31flk2 ligand (FL), and/or stem cell factor (SCF), interleukin 6 (IL-6), interleukin 3 (IL-3) under serum-free conditions. After I or 2 weeks of culture, cells were examined for clonogenic progenitors and percentage of CD34+ CD38- cells. In the presence of TPO, FL, and SCF, fetal MSC cells supported more than a 35- and 20-fold expansion of CD34+ cells and colonyforming units in culture after 1 and 2 weeks of incubation, respectively. In addition, LTC-IC assay were expanded more than 7- and 16-fold after 1 and 2 weeks of culture, respectively. UCB-HSC can be expanded in culture to numbers theoretically adequate for safe, rapid engraftment of adult patients. Additional studies are needed to establish the functional activity of expanded UCB-HSC. This ex vivo expansion system should prove valuable in clinical settings in which stromal cells are available from recipients or stem cell donors.

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 Primary and secondary malignant central nervous system (CNS) tumors are devastating invasive tumors able to give rise to many kinds of differentiated tumor cells. Glioblastoma multiform (GBM), is the most malignant brain tumor, in which its growth and persistence depend on cancer stem cells with enhanced DNA damage repair program that also induces recurrence and resists current chemo- and radiotherapies. Unlike non-tumor stem cells, tumor stem cells lack the normal mechanisms that regulate proliferation and differentiation, resulting in uncontrolled production and incomplete differentiation of tumor cells. In current paper recent developments and new researches in the field of brain tumor stem cells have been reviewed.

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Background: Doxorubicin, by aggregating in bone marrow, causes genotoxic effects, and thus reduces the repair ability of cells. The present study was conducted as an in vitro evaluation of age effects on the cytotoxicity induced by doxorubicin in mesenchymal stem cells (MSCs).
   Methods: The MSCs of female BALB/c mice aged 1, 8, and 16 months were separated, characterized, and subsequently evaluated in cellular growth media. After 24 hours, exposure of the MSCs of the 3 groups of mice to doxorubicin (25, 50, 100, 200, 400, 800, 1200 nM) and cytotoxicity were assessed, and the sublethal dose was determined using flow cytometry technique and lactate dehydrogenase (LDH) release assay.
   Results: The IC50 values determined by flow cytometry for the separated MSCs of 1 young, 8 middle- aged, and 16 old mice were and respectively. Interestingly, the results of these 2 methods in determining cytotoxicity were in agreement, and a concentration of approximately 25 nM was considered to be the shared sublethal dose for different ages.
   Conclusion: The results indicated that MSCs of middle-aged mice were more resistant to the toxic effects of the drug. Besides, MSCs separated from the old mice were the most sensitive to chemotherapy and its side effects such as disruptions of cell proliferation and viability. These disruptions can be ascribed to the alteration of function and physiological processes with age. Determining proper concentration of doxorubicin drug to destruct cancerous cells based on age and individual sensitivity can minimize the amount of toxicity.
 

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Cancer stem cells (CSCs) have critical roles in tumor development, progression, and recurrence. They are responsible for current cancer treatment failure and remain questionable for the design and development of new therapeutic strategies. With this issue, medical imaging provides several clues for finding biological mechanisms and strategies to treat CSCs. This review aims to summarize current molecular imaging approaches for detecting CSCs. In addition, some promising issues for CSCs finding and explaining biological mechanisms have been addressed. Among the molecular imaging approaches, modalities including Magnetic resonance imaging (MRI) and positron emission tomography (PET) have the greatest roles and several new approaches such as optical imaging are in progress.

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Background: Ischemic cardiomyopathies are the leading causes of mortality and morbidity. Stem cell therapy using amniotic membrane mesenchymal stem cells have emerged as a promising cardiac regeneration modality. They have shown great immunological advantage when used in allogeneic or xenogeneic transplantation. The aim of the current study is to accumulate evidence from published preclinical studies on the application of amniotic membrane derived mesenchymal stem cells (AMSCs) in the treatment of ischemic cardiomyopathies including myocardial ischemia and heart failure. The aim is to define if there is enough high-quality current evidence to support starting the use of these cells in clinical trials.
   Methods: PubMed, SCOPUS, EMBASE, and ISI Web of Science databases were searched without temporal and language restrictions. Data were extracted from selected studies. The primary outcomes were left ventricular ejection fraction (LVEF) and LV fibrosis. The risk of bias (ROB) assessment was performed using SYRCLE’s ROB tool. After qualitative synthesis, provided that data meets the criteria for quantitative analysis, a meta-analysis was performed using Stata software V12 to investigate the heterogeneity of the data and to get an overall estimate of the effect size of the treatment on each outcome.
   Results: On primary search, 438 citations were retrieved. After screening, three studies were selected for quantitative analysis of each of the outcomes LVEF and LV fibrosis. Their administration in acute and chronic MI alleviates heart failure and improves LVEF (SMD=3.56, 95% CI: 2.24-4.87, I-squared=83.1%, p=0.003) and reduces infarct size (SMD= -4.41, 95% CI: (-5.68)-(-3.14), I-squared=79.0%, p=0.009). These observations were achieved in the acute MI model, HF following ischemia due to coronary artery stenosis and coronary artery occlusion with the early restoration of the perfusion.
   Conclusion: Present low and medium quality evidence from preclinical studies confirm the efficacy of the AMSCs in the preclinical models of acute MI and HF following ischemia due to coronary artery stenosis and permanent/temporary coronary artery occlusion. High-quality preclinical studies are indicated to bridge the gaps in translation of the current findings of AMSCs research for the treatment of patients with acute and chronic myocardial ischemia and heart failure.

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Background: GD2 synthase (GD2S) is the key enzyme required for ganglioside GD2 synthesis. It is commonly expressed in normal tissues and various cancers. Ganglioside GD2 is identified as a breast cancer stem cells (BCSCs) marker that promotes tumorigenesis. As GD2S has been found to be a useful molecular marker in neuroblastoma and retinoblastoma tumors, we suggest that it can be considered as a suitable candidate for the detection of CSCs in breast cancer tissues.
    Methods: Expression of GD2S was examined in 65 breast tumors compared to adjacent normal tissues, applying quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The association between GD2S expression level and patients’ clinical characteristics was also assessed.
   Results: Our findings showed that GD2S mRNA expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissue samples (4.92-fold change, p<0.001) in advanced grades (p<0.001) and stages (p<0.001). It was also shown that GD2S protein expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissues (4.86-fold change, p=0.010) in advanced grades (p=0.010), stages (p=0.005) and larger tumor size (p=0.002).
   Conclusion: The current study showed that increased expression of GD2S in advanced breast cancer potentiates it as a promising tumor marker in these patients.

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    Background: Currently, stem cell therapy has been proposed as an efficient strategy to prevent or treat myocardial injuries. The current study was conducted to examine cardioprotective effects of human mesenchymal stem cells derived from amniotic membrane (hAMSCs) against isoproterenol (ISO)-induced myocardial injury and explore its potential mechanisms.
    Methods: The hAMSCs were injected intramyocardially in male Wistar rats 28 days after last injection of ISO (170 mg/kg body weight for 4 consecutive days). The echocardiography was performed to confirm induction of myocardial damage and cardiac function 28 days after last injection of ISO and 4 weeks hAMSCs transplantation after HF induction. The expression of apoptotic markers such as Bcl-2, Bax and P53 was evaluated using Western blotting assay. Masson’s trichrome staining was used to determine fibrosis. The cytoarchitecture of myocardial wall and morphology of cells were investigated using hematoxylin and eosin (H&E) staining.
   Results: As compared to ISO group, hAMSCs transplantation after heart failure (HF) induction significantly blunted the increasing of cardiac dimensions and restored ejection fraction (EF) and fractional shortening (FS) parameters (p<0.05). Moreover, hAMSCs transplantation after HF induction increased the expression of antiapoptotic markers such as Bcl-2 and decreased the expression of pro-apoptotic markers such as P53 and Bax (p<0.05). As compared to ISO group, hAMSCs transplantation after HF induction markedly reduced interstitial myocardial fibrosis and contributed to maintain of normal cytoarchitecture of myocardial wall and morphology of cells.
   Conclusion: Collectively, the results of current study suggest that transplantation of hAMSCs confers cardioprotection by targeting ISO‐induced mitochondria‐dependent (intrinsic) pathway of apoptosis.

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    Background: The burn wound is one of the health problems in the world that affects physical and mental health. Today, adipose-derived mesenchymal stem cells (ADSCs) have received medical attention for their accessibility and the ability to reproduce and repair. The present study was designed to investigate the effect of ADSCs on burn wound healing.
   Methods: The present experimental study was performed on 36 male Wistar rats divided into 1 control group and 3 experimental groups. The second-degree burns with a radius of 10 mm were induced after anesthesia. ADSCs and Dulbecco's Modified Eagle Medium (DMEM) were injected into the dermis around the burn area in the ADSCs and DMEM groups, respectively. Silver sulfadiazine (SSD) ointment was applied topically once daily as the SSD group. The control group did not receive any treatment. The rats were evaluated for 21 days. Wound healing rate, histopathological parameters, and the number of fibroblasts were evaluated by the immunofluorescence technique and vascular endothelial growth factor and transforming growth factor β (TGF-β) gene expression by reverse transcription-polymerase chain reaction. The results were entered into SPSS software (SPSS Inc) and analyzed with 1-way analysis of variance and repeated measures analysis.
   Results: The number of fibroblasts, the number of vessels, TGF-β, and VEGF gene expression in the burn area were significantly higher in the ADSCs group than in the SSD, DMEM, and control groups. The results also showed that the amount of inflammation was significantly lower in the ADSCs group compared with the control group (p<0.001). Moreover, the percentage of wound recovery was significantly higher in the ADSCs group compared with other groups (p<0.001).
   Conclusion: ADSCs accelerate and improve burn wound healing by affecting fibroblasts, keratinocytes, and inflammatory cells as well as increasing the expression of the TGF-β and VEGF genes, and thus increase in angiogenesis.

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Background: In patients with diabetes, transplantation of stem cells increases C-peptide levels and induces insulin independence for some period. Today, this positive therapeutic outcome is widely attributed to the well-documented immunomodulatory properties of stem cells. The aim of this study was to report alternations (the trend of increase or decrease) in different lymphocyte populations in a stem cell clinical trial performed in our institute.
   Methods: Recorded data of a clinical trial conducted on 72 patients with type 1 diabetes who had received fetal stem cell transplantation several years ago and were regularly monitored before and after the procedure in 1, 3, 6, 12, 24 months were analyzed. In these regular follow-up visits, insulin demand, HbA1c, C-peptide, and alternation to B cell and T cell populations were analyzed and recorded. For the purpose of the current study, patients were retrospectively divided into 2 groups, namely, those with the positive response to treatment and patients without such response. Temporary positive therapeutic response was defined by 2 different indicators, namely, plasma C-peptide levels and insulin dose-adjusted A1C (IDAA1c), which was calculated as A1C (percent) + (4 × insulin dose (units per kilogram per 24 h). Data analysis was performed by means of SPSS Version 18.
   Results: Besides the short-term therapeutic effect, we observed remarkably significant alternations to the populations of B and T lymphocytes in the recipients. When patients were retrospectively assigned to 2 different groups of patients with a positive therapeutic response (based on C-peptide changes) and those without it, it was observed that alternations to different populations of B-cells and T-cells were significantly different in these 2 groups.
   Conclusion: Our results demonstrated that transplantation of stem cells leads to significant positive therapeutic outcomes in one group of patients who showed totally distinct patterns of alternation to different groups of lymphocytes. 

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Background: The quality of the wound healing at the donor site significantly determines the overall condition of the burn patient, the extent of wound fluid and protein losses, the severity of any systemic in-flammatory reaction, and the intensity of the pain syndrome. It is known that the stromal vas-cular fraction (SVF)  has a beneficial effect on the healing of wound defects. This study is aimed at assessing the safety and effectiveness of the application of the SVF of autologous adipose tis-sue to stimulate wound healing of the donor site in patients with burns.
   Methods: This placebo-controlled clinical study included 38 patients with third-degree thermal skin burns. The patients underwent liposuction, enzymatic isolation of the SVF, and intradermal injection of the preparation into the wounds in the donor site, followed by tewametry, cutome-try, thermography and biopsy after 12 days. Quantitative indicators were compared using the Mann-Whitney test for unrelated groups and the Wilcoxon test for related groups. Spearman's rank correlation coefficient (RS) was used to assess the correlation
   Results: Epithelization of the wounds in all patients was seen over an average area of 88 (84;92) %, there being no significant differences between the actual and the control wound sites for this parameter. Transdermal water loss in the test wound sites was 2 times lower than in the control sites (P = 0.001). The wound donor sites regained their temperature distribution faster than the control sites (P = 0.042). Histological preparations of the skin of the wound sites revealed that their epidermal layer was 19% thicker compared to the controls (P = 0.043). It should be noted that five adverse events related to manipulations in the postoperative period were registered.
   Conclusions: Transplantation of SVF autologous adipose tissue into the wound area in most clinical cases proceeded without complications. The area of epithelialization of wound areas af-ter the introduction of SVF did not change, although a significant decrease in transdermal water loss was observed in the wound areas with an improvement in their thermoregulation and an increase in the thickness of the epidermis.
 

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Background: This study aims to map the research trends in the field of stem cell research in Iran by presenting a systematic and analytical bibliometrics approach based on data from the Web of Science database.
   Methods: In this study, we provide a visualization overview of the distribution of stem cell publications in Iran. The HistCite software was used to draw and analyze the historiographical maps, based on Global Citation Score (GSC) and Local Citation Score (LCS) in order to indicate the most frequent thematic trends. The accuracy of clustering and classification of scientific fields is enhanced by the incorporation of algorithms and main bibliometric analysis.
   Results: A total of 5123 records were collected from the Web of Science database in 2020. The most prolific author had a GCS of 5890 and the most productive university earned GCS of 13677. “Cell Journal,” with 186 records contributed the highest number of publications. The highest cited document based on GSC had a score of 646 and the highest cited article based on LCS had a score of 71. We documented regular growth in outputs. In addition, the scientific maps based on LCS and GCS have been drawn. The prominent, distinguished areas of study revolve around differentiation, generation, proliferation, and the therapeutic use of stem cells as well as “genotoxicity in stem cells”, “mesenchymal stem cells” and “embryonic stem cells”.  Journal articles were the predominant document type.
   Conclusion: Research on stem cells is a biomedical venture with great scientific impact, and its development in Iran is undeniable. This study provides an overview and a framework for the weaknesses and strengths of Iranian research outputs on stem cells, representing the main clusters in scientific maps. We hope that our results help researchers to plan future studies and promote their research productions.