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Cell Journal [Yakhteh]. 2019; 20 (4): 450-458
in English | IMEMR | ID: emr-199613


Objective: Mesenchymal stem cells [MSC] from various sources have the potentials to positively affect regenerative medicine.Furthermore, pre-conditioning strategies with different agents could improve the efficacy of cell therapy. This study compares the effects of an anti-inflammatory and antioxidant agent, melatonin, on protection of bone marrow-derived MSCs [BMSCs] and adipose tissue-derived MSCs [ADSCs]

Materials and Methods: In this experimental study, rat BMSCs and ADSCs were isolated and expanded. Pre-conditioning was performed with 5 ìM melatonin for 24 hours. Cell proliferation and viability were detected by MTT assay. Expression of BAX, BCL2, melatonin receptors and osteocalcin genes were evaluated by reverse transcriptase-polymerase chain reaction [RT-PCR]. Also, apoptosis was detected with tunnel assay. Osteogenic differentiation was analyzed using alizarin red staining

Results: No significant increase was found in cell viability between BMSCs and ADSCs after melatonin preconditioning. Following melatonin preconditioning, BAX expression was significantly down-regulated in both ADSCs and BMSCs [P<0.05], with the difference being more significant in ADSCs compared to BMSCs. BCL2 expression was increased significantly in both cell types after preconditioning. Metalothionine 1 and Metalothionine 2 were both upregulated significantly in the two cell types [P<0.05]. Melatonin increased osteogenesis capability through increasing osteocalcin expression. However, expression of osteocalcin in BMSCs before and after preconditioning was higher than that in ADSCs. On the other hand, melatonin expression in ADSCs was in higher levels than in BMSCs. Melatonin also improved alizarin red concentration significantly in both BMSCs and ADSCs [P<0.05]. Alizarin red staining severity increased significantly in ADSCs after preconditioning compared to BMSCs [P<0.05]

Conclusion: Here we have shown that the effects of preconditioning on melatonin expression in ADSCs are higher than those in BMSCs. These findings could be used in adoption of a proper preconditioning protocol based on the sources of MSCs in specific clinical applications, especially in bone regeneration

Cell Journal [Yakhteh]. 2017; 19 (3): 361-374
in English | IMEMR | ID: emr-193044


Objective: Immunotherapy and gene therapy play important roles in modern medicine. The aim of this study is to evaluate the overexpression of interleukin-4 [IL-4], IL-10 and leukemia inhibitory factor [LIF] in Wharton's jelly stem cells [WJSCs] in the experimental autoimmune encephalomyelitis [EAE] mice model

Materials and Methods: In this experimental study, a DNA construction containing IL- 4, IL-10 and LIF was assembled to make a polycistronic vector [as the transfer vector]. Transfer and control vectors were co-transfected into Human Embryonic Kidney 293 [HEK-293T] cells with helper plasmids which produced recombinant lentiviral viruses [rLV]. WJSCs were transduced with rLV to make recombinant WJSC [rWJSC]. In vitro protein and mRNA overexpression of IL-4, LIF, and IL-10 were evaluated using quantitative polymerase chain reaction [qPCR], enzyme-linked immunosorbent assay [ELISA] and western blot [WB] analysis. EAE was induced in mice by MOG-CFA and pertussis toxin. EAE mice were injected twice with 2×10[5] rWJSCs. The in vivo level of IL-4, LIF, IL-10 cytokines and IL-17 were measured by ELISA. Brain tissues were analyzed histologically for evaluation of EAE lesions

Results: Isolated WJSCs were performed to characterize by in vitro differentiation and surface markers were analyzed by flow cytometry method. Cloning of a single lentiviral vector with five genes was done successfully. Transfection of transfer and control vectors were processed based on CaPO4 method with >90% efficiency. Recombinant viruses were produced and results of titration showed 2-3×10[7] infection-unit/ml. WJSCs were transduced using recombinant viruses. IL-4, IL-10 and LIF overexpression were confirmed by ELISA, WB and qPCR. The EAE mice treated with rWJSC showed reduction of Il-17, and brain lesions as well as brain cellular infiltration, in vivo. Weights and physical activity were improved in gene-treated group

Conclusion: These results showed that gene therapy using anti-inflammatory cytokines can be a promising approach against multiple sclerosis [MS]. In addition, considering the immunomodulatory potential of WJSCs, an approach using a combination of WJSCs and gene therapy will enhance the treatment efficacy

Cell Journal [Yakhteh]. 2016; 17 (4): 639-647
in English | IMEMR | ID: emr-179292


Objective: Bone marrow [BM] is one of the major hematopoietic organs in postnatal life that consists of a heterogeneous population of stem cells which have been previously described. Recently, a rare population of stem cells that are called very small embryonic-like [VSEL] stem cells has been found in the BM. These cells express several developmental markers of pluripotent stem cells and can be mobilized into peripheral blood [PB] in response to tissue injury. In this study we have attempted to investigate the ability of these cells to migrate toward an injured spinal cord after transplantation through the tail vein in a rat model

Materials and Methods: In this experimental study, VSELs were isolated from total BM cells using a fluorescent activated cell sorting [FACS] system and sca1 and stage specific embryonic antigen [SSEA-1] antibodies. After isolation, VSELs were cultured for 7 days on C2C12 as the feeder layer. Then, VSELs were labeled with 1,1´-dioctadecyl-3,3,3´,3´- tetramethylindocarbocyanine perchlorate [DiI] and transplanted into the rat spinal cord injury [SCI] model via the tail vein. Finally, we sought to determine the presence of VSELs in the lesion site

Results: We isolated a high number of VSELs from the BM. After cultivation, the VSELs colonies were positive for SSEA-1, Oct4 and Sca1. At one month after transplantation, real-time polymerase chain reaction analysis confirmed a significantly increased expression level of Oct4 and SSEA-1 positive cells at the injury site

Conclusion: VSELs have the capability to migrate and localize in an injured spinal cord after transplantation

Zahedan Journal of Research in Medical Sciences. 2014; 16 (6): 12-15
in English | IMEMR | ID: emr-169273


Increased vascular stiffness is a marker of atherosclerosis, which is diagnosed in the early stages of diabetes II. Matrix metalloproteinases [MMPs] and their tissue inhibitors [TIMPs] are a family of proteolytic enzymes necessary for structure and function of great vessels. This study examined the effects of 8 weeks of aerobic exercise on MMP9 and TIMP-1 levels in type II diabetic women. This is a quasi-experimental study which included 20 in type II diabetic women with mean age of 53.2 +/- 2.5 years, body mass index [BMI] of 28.73 +/- 2.27 and fat percentage of 30.6 +/- 2.05, who were randomly divided into two groups: aerobic exercise group [8 weeks, 3 sessions per week for 50 minutes] and control group. To examine changes in MMP[9] and TIMP-1, 5 ml of blood was taken from the brachial vein of patients before and 48 hours after completion of exercise period and after 12 hours of fasting at rest. Data analysis was performed using SPSS-16 software with the independent and paired t-tests. A significant decrease was observed in body mass index and body fat percentage in the experimental group [p<0.05]. Compared with the control group, the aerobic exercise group showed a significant decrease in MMP[9] [p=0.01] and a significant increase in TIMP-1 levels [p=0.02] after 8 weeks of aerobic exercise. The results showed that aerobic exercise as a stimulus can change the levels of matrix metalloproteinases and their tissue inhibitors in order to prevent cardiovascular diseases in diabetics

Cell Journal [Yakhteh]. 2012; 14 (3): 231-236
in English | IMEMR | ID: emr-153862


Ecstasy, also known as 3, 4-methylenedioxymethamphetamine [MDMA], is a psychoactive recreational hallucinogenic substance and a major worldwide recreational drug. There are neurotoxic effects observed in laboratory animals and humans following MDMA use. MDMA causes apoptosis in neurons of the central nervous system [CNS]. Withdrawal signs are attenuated by treatment with the adenosine receptor [A2A receptor]. This study reports the effects of glutamyl cysteine synthetase [GCS], as an A2A receptor agonist, and succinylcholine [SCH], as an A2A receptor antagonist, on Sprague Dawley rats, both in the presence and absence of MDMA. In this experimental study, we used seven groups of Sprague Dawley rats [200-250 g each]. Each group was treated with daily intraperitoneal [IP] injections for a period of one week, as follows: i. MDMA [10 mg/kg]; ii. GCS [0.3 mg/kg]; iii. SCH [0.3 mg/kg]; iv. GCS + SCH [0.3 mg/kg each]; v. MDMA [10 mg/kg] + GCS [0.3 mg/kg]; vi. MDMA [10 mg/kg] + SCH [0.3 mg/kg]; and vi. normal saline [1 cc/kg] as the sham group. Bax [apoptotic protein] and Bcl-2 [anti-apoptotic protein] expressions were evaluated by striatum using RT-PCR and Western blot analysis. There was a significant increase in Bax protein expression in the MDMA+SCH group and a significant decrease in Bcl-2 protein expression in the MDMA+SCH group [p<0.05]. A2A receptors have a role in the apoptotic effects of MDMA via the Bax and Bcl-2 pathways. An agonist of this receptor [GCS] decreases the cytotoxcity of MDMA, while the antagonist of this receptor [SCH] increases its cytotoxcity

Animals, Laboratory , N-Methyl-3,4-methylenedioxyamphetamine/adverse effects , Receptors, Purinergic P1 , Adenosine A2 Receptor Agonists , Adenosine A2 Receptor Antagonists , Cellular Apoptosis Susceptibility Protein , Receptors, Adenosine A2
IJI-Iranian Journal of Immunology. 2012; 9 (2): 119-127
in English | IMEMR | ID: emr-152168


Genistein [GEN], a naturally occurring flavonoid present in soy bean, has attracted scientific interest for its possible benefits in cancer. The potential immunomodulatory effects of genistein on the immune system and against TC-1 tumor cell line were evaluated in adult female C57BL/6 mice. Mice were treated with GEN 10 days before to 10 days after the tumor induction. Thirty days after the last GEN treatment, lymphocyte proliferation, Lactase Dehydrogenase [LDH] cytolytic activity and cytokine secretion were analyzed in GEN and control groups. The results showed that ingestion of genistein significantly increased lymphocyte proliferation and LDH release. Furthermore, the treatment with genistein also caused a significant increment in interferon gamma [IFN-alpha]. In addition, the treatment achieved significant therapeutic effect in tumor models compared to the control group. These results indicated that the effect of GEN on tumor growth may be attributed to its effect on lymphocyte proliferation, cytolytic activity and IFN-alpha production. These results demonstrate that GEN exerts an immunomodulatory effect in a mouse model of Human Papillomavirus [HPV] associated-cervical cancer