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Cell Journal [Yakhteh]. 2019; 20 (4): 477-482
in English | IMEMR | ID: emr-199616


Objective: Type 1 diabetes is caused by destruction of beta cells of pancreas. Vildagliptin [VG], a dipeptidyl peptidase IV [DPP IV] inhibitor, is an anti-diabetic drug, which increases beta cell mass. In the present study, the effects of VG on generation of insulin-producing cells [IPCs] from adipose-derived mesenchymal stem cells [ASCs] is investigated

Materials and Methods: In this experimental study, ASCs were isolated and after characterization were exposed to differentiation media with or without VG. The presence of IPCs was confirmed by morphological analysis and gene expression [Pdx-1, Glut-2 and Insulin]. Newport Green staining was used to determine insulin-positive cells. Insulin secretion under different concentrations of glucose was measured using radioimmunoassay method

Results: In the presence of VG the morphology of differentiated cells was similar to the pancreatic islet cells. Expression of Pdx-1, Glut-2 and Insulin genes in VG-treated cells was significantly higher than the cells exposed to induction media only. Insulin release from VG-treated ASCs showed a nearly 3.6 fold [P<0.05] increase when exposed to a highglucose medium in comparison to untreated ASCs. The percentage of insulin-positive cells in the VG-treated cells was approximately 2.9-fold higher than the untreated ASCs

Conclusion: The present study has demonstrated that VG elevates differentiation of ASCs into IPCs. Improvement of this protocol may be used in cell therapy in diabetic patients

Article in English | WPRIM | ID: wpr-732287


Background: Recent studies have demonstrated that many nanoparticles have an adverseor toxic effect on the kidney.Objective: To investigate the nephroprotective effect of quercetin (QT) against renal injuryinduced by titanium dioxide nanoparticles (NTiO2) in rats.Methods: NTiO2-intoxicated rats received 50 mg/kg of NTiO2 for seven days. The QT +NTiO2 group was pretreated with QT for seven days before being administered NTiO2. Uric acid,creatinine, and blood urea nitrogen were considered to be biomarkers of nephrotoxicity. Catalase(CAT) and superoxide dismutase (SOD) activities and renal levels of malondialdehyde (MDA) weremeasured to assess the oxidative stress caused by NTiO2.Results: NTiO2 significantly increased the plasma level of the biomarkers. It alsosignificantly decreased the activities of CAT (P = 0.008) and SOD (P = 0.004), and significantlyincreased the MDA levels (P = 0.007). NTiO2 caused proximal tubule damage, the accumulationof red blood cells, the infiltration of inflammatory cells, and reduced the glomerular diameters,as well as induced apoptosis in the proximal tubules. Pre-treatment with QT attenuated thehistological changes, normalised the plasma biomarkers, suppressed oxidative stress, amelioratedthe activities of CAT (P = 0.007) and SOD (P = 0.006), and reduced apoptosis (P < 0.001).Conclusion: QT was found to have a potent protective effect against nephrotoxicityinduced by NTiO2 in rats. It also reduced apoptosis caused by NTiO2.

Cell Journal [Yakhteh]. 2016; 17 (4): 720-729
in English | IMEMR | ID: emr-179300


Objective: To evaluate the effect of Exendine-4 [EX-4], a Glucagon-like peptide 1 [GLP-1] receptor agonist, on the differentiation of insulin-secreting cells [IPCs] from rat adipose-derived mesenchymal stem cells[ADMSCs]

Materials and Methods: In this experimental study, ADMSCs were isolated from rat adipose tissue and exposed to induction media with or without EX-4. After induction, the existence of IPCs was confirmed by morphology analysis, expression pattern analysis of islet-specific genes [Pdx-1, Glut-2 and Insulin] and insulin synthesis and secretion

Results: IPCs induced in presence of EX-4 were morphologically similar to pancreatic islet-like cells. Expression of Pdx-1, Glut-2 and Insulin genes in EX-4 treated cells was significantly higher than the cells exposed to differentiation media without EX-4. Compared to EX-4 untreated ADMSCs, insulin release from EX-4 treated ADMSCs showed a nearly 2.5 fold [P<0.05] increase when exposed to a high glucose [25 mM] medium. The percentage of insulin positive cells in the EX-4 treated group was approximately 4-fold higher than in the EX-4 untreated ADMSCs

Conclusion: The present study has demonstrated that EX-4 enhances the differentiation of ADMSCs into IPCs. Improvement of this method may help the formation of an unlimited source of cells for transplantation

Cell Journal [Yakhteh]. 2016; 18 (3): 289-301
in English | IMEMR | ID: emr-183763


Objective: the organotypic co-culture is a well-known technique to examine cellular interactions and their roles in stem cell proliferation and differentiation. This study aims to evaluate the effects of dermal fibroblasts [DFs] on epidermal differentiation of adipose-derived stem cells [ASCs] using a three-dimensional [3D] organotypic co-culture technique

Materials and Methods: in this experimental research study, rat DFs and ASCs were isolated and cultured separately on electrospun polycaprolactone [PCL] matrices. The PCL matrices seeded by ASCs were superimposed on to the matrices seeded by DFs in order to create a 3D organotypic co-culture. In the control groups, PCL matrices seeded by ASCs were placed on matrices devoid of DFs. After 10 days, we assessed the expressions of keratinocyte-related genes by real-time reverse transcriptase-polymerase chain reaction [RT-PCR] and expression of pan-cytokeratin protein by immunofluorescence in the differentiated keratinocyte-like cells from co-culture and control groups. Keratinocyte-like cell morphologies were also observed by scanning electron microscopy [SEM]

Results: the early, intermediate, and terminal differentiation keratinocyte markers-Cytokeratin14, Filaggrin, and Involucrin significantly expressed in the co-culture groups compared to the control ones [P<0.05]. We observed pan-cytokeratin in keratinocyte-like cells of both groups by immunofluorescence. SEM observation of the co-culture groups showed that the differentiated keratinocyte-like cells developed a polygonal cobblestone shape, considered characteristic of keratinocytes

Conclusion: the 3D organotypic co-culture bilayered construct that consisted of DFs and ASCs was an effective technique for epidermal differentiation of ASCs. This co-culture might be useful for epidermal differentiation of stem cells for future applications in skin regeneration

Article in English | WPRIM | ID: wpr-78764


PURPOSE: The present study was conducted to evaluate the favorable or harmful effects of betulinic acid (BA) on a diabetic reproductive system. MATERIALS AND METHODS: In this experimental study, 60 male Naval Medical Research Institute mice (20∼25 g) were randomly divided into 6 groups: control, diabetes, diabetes+BA (10, 20, and 40 mg/kg), and diabetes+ metformin (200 mg/kg). A diabetic model was induced by a single dose of streptozotocin (STZ) (65 mg/kg) injection intraperitoneally 15 minutes after an intraperitoneal administration of nicotinamide (NA) (120 mg/kg). BA and metformin were gavaged for 2 weeks after confirmed diabetes induction in the treatment groups. One day after the last treatment, plasma luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone levels were evaluated. The cauda epididymis and testis were removed to analyze the sperm count and testis histopathology. RESULTS: LH levels increased in diabetic (p<0.001) and diabetic BA-treated mice (p=0.009). Plasma levels of testosterone (p< 0.001) and sperm count (p=0.04) decreased in these groups when compared to the control group. Furthermore, administration of 10 mg/kg (p=0.001), 20 mg/kg (p=0.004), or 40 mg/kg (p<0.001) of BA led to a greater reduction in plasma testosterone levels compared to the diabetes group. Seminiferous tubule vacuole numbers increased in diabetic and diabetic BA-treated mice, but testis morphology and FSH level assessment revealed no significant differences between the groups. CONCLUSIONS: STZ-NA can induce diabetic alterations in the male reproductive system and the administration of BA in diabetic treated mice resulted in a worse outcome.

Academies and Institutes , Animals , Diabetes Mellitus , Epididymis , Follicle Stimulating Hormone , Humans , Luteinizing Hormone , Male , Metformin , Mice , Niacinamide , Plasma , Seminiferous Tubules , Sperm Count , Spermatozoa , Streptozocin , Testis , Testosterone , Vacuoles
Cell Journal [Yakhteh]. 2015; 17 (3): 412-421
in English | IMEMR | ID: emr-174874


Objective: Zinc oxide nanoparticles [ZnO-NPs] are increasingly used in sunscreens, biosensors, food additives, pigments, manufacture of rubber products, and electronic materials. There are several studies about the effects of NPs on dermal fibroblast or keratinocytes, but very little attention has been directed towards adipose-derived mesenchymal stem cells [ASCs]. A previous study has revealed that ZnO-NPs restricted the migration capability of ASCs. However, the potential toxicity of these NPs on ASCs is not well understood. This study intends to evaluate the effects of ZnO-NPs on subcutaneous ASCs

Materials and Methods: In this experimental study, In order to assess toxicity, we exposed rat ASCs to ZnO-NPs at concentrations of 10, 50, and 100 Mug/ml for 48 hours. Toxicity was evaluated by cell morphology changes, cell viability assay, as well as apoptosis and necrosis detection

Results: ZnO-NPs concentration dependently reduced the survival rates of ASCs as revealed by the trypan blue exclusion and 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium-bromide [MTT] tests. ZnO-NPs, at concentrations of 10 and 50 Mug/ml, induced a significant increase in apoptotic indices as shown by the annexin V test. The concentration of 10 Mug/ml of ZnO-NPs was more toxic

Conclusion: Lower concentrations of ZnO-NPs have toxic and apoptotic effects on subcutaneous ASCs. We recommend that ZnO-NPs be used with caution if there is a dermatological problem

IJPR-Iranian Journal of Pharmaceutical Research. 2015; 14 (1): 329-334
in English | IMEMR | ID: emr-154894


Grape seed proanthocyanidin extract [GSPE] has a broad spectrum of biologic properties against oxidative stress. This study aimed to investigate the effects of GSPE on biochemical factors and antioxidant enzymes of erythrocyte in diabetic rats. Diabetes was induced through single injection of streptozotocin [50 mg.Kg[-1], i.p]. Forty Male Sprague-Dawley rats were divided into four Groups: Group 1, healthy control group; Group 2, healthy group treated with GSPE [200 mg.Kg[-1]]; Group 3, diabetic control group; Group 4, diabetic group treated with GSPE [200 mg.Kg[-1]] for 4 weeks. At the end, the experimental animals were sacrificed and blood samples were collected and plasma parameters and erythrocytes antioxidant status were evaluated. The results show, treatment with GSPE significantly reduced [P<0.001] urine volume, proteinuria and biochemical factors such as blood urea nitrogen, creatinine, triglyceride, total cholesterol, low density lipoprotein and very low density lipoprotein as well as malondialdehyde. Also GSPE treatment significantly [P<0.005] increased high density lipoprotein, total protein and albumin. Moreover GSPE significantly increased antioxidant enzymes activity such as: superoxide dismutase, glutathione peroxidase and catalase. These results suggest that GSPE can ameliorate biochemical abnormalities and antioxidant system status in streptozotocin- induced diabetic rats probably by its potent antioxidant features

Animals, Laboratory , Grape Seed Extract , Antioxidants , Erythrocytes , Rats, Sprague-Dawley , Diabetes Mellitus, Experimental , Streptozocin , Biomarkers
Cell Journal [Yakhteh]. 2014; 16 (1): 43-52
in English | IMEMR | ID: emr-148446


Finding cell sources for cartilage tissue engineering is a critical procedure. The purpose of the present experimental study was to test the in vitro efficacy of the beta-tricalcium phosphate-alginate-gelatin [BTAG] scaffold to induce chondrogenic differentiation of human umbilical cord blood-derived unrestricted somatic stem cells [USSCs]. In this experimental study, USSCs were encapsulated in BTAG scaffold and cultured for 3 weeks in chondrogenic medium as chondrogenic group and in Dulbecco's Modified Eagle's Medium [DMEM] as control group. Chondrogenic differentiation was evaluated by histology, immunofluorescence and RNA analyses for the expression of cartilage extracellular matrix components. The obtain data were analyzed using SPSS version 15. Histological and immunohistochemical staining revealed that collagen 2 was markedly expressed in the extracellular matrix of the seeded cells on scaffold in presence of chondrogenic media after 21 days. Reverse transcription-polymerase chain reaction [RT-PCR] showed a significant increase in expression levels of genes encoded the cartilage- specific markers, aggrecan, type 1 and 2 collagen, and bone morphogenetic protein [BMP]-6 in chondrogenic group. This study demonstrates that BTAG can be considered as a suitable scaffold for encapsulation and chondrogenesis of USSCs

Humans , Chondrogenesis , Calcium Phosphates , Fetal Blood , Alginates , Mesenchymal Stem Cells
IJRM-International Journal of Reproductive Biomedicine. 2013; 11 (2): 119-126
in English | IMEMR | ID: emr-193216


Background: galectin-3 [Gal-3], a beta-galactoside-binding lectin, is a multifunctional lectin that involves in a number of critical biological processes

Objective: the purpose of this study was to investigate the expression pattern of Gal-3 in mouse endometrium during estrus phase of estrous cycle and pre-implantation

Materials and Methods: in this experimental study 42 NMRI female mice were divided in seven different groups. Ovulation in NMRI female mice was stimulated by injecting hMG and hCG. Estrus phase was considered as stimulated and un-stimulated groups. The other groups of mice were mated, and the day of vaginal plug formation was considered as the day 1 of pregnancy. The mice of all groups were sacrificed on different days of pre-implantation period and their uterine horns were fixed and avid in- biotin complex method of immunohistochemistry [IHC] was applied

Results: in estrus group, Gal-3 immunoreactivity in luminal epithelium was strong, in stromal cells very strong, in glandular epithelium very weak and endothelial cells very strong. No identifiable difference was observed in un-stimulated and stimulated estrus phase. In test groups, days 1-2, insignificant difference of Gal-3 expression was observed. On day 3, luminal epithelium and stromal cells showed significant decrease in comparison to estrus and day 1 [p=0.001]. On the 4[th] and 5[th] days, luminal epithelium and stromal cells showed significant decrease in comparison to estrus phase and days 1-3 [p=0.0001]

Conclusion: the data suggested that successful implantation is probably associated with the downregulation of Gal-3 in the mouse endometrium at the beginning of pregnancy