Wound healing activity of Pimpinella anisum methanolic extract in streptozotocin-induced diabetic rats

Objective: To assess the wound healing potential of Pimpinella anisum on cutaneous wounds in diabetic rats. Method: Full-thickness excisional wounds were made on the back of male, Sprague-Dawley rats with diabetes. The rats were randomly allocated into four treatment groups: 1 ml basal cream; tetracycline (3); Pimpinella anisum 10 for 14 days; and a control group. At days seven, 14 and 21 post-injury, five animals of each group were euthanised, and wounds were assessed through gross, histopathological and oxidant/antioxidant evaluations. Additionally, the dry matter and hydroxyproline contents of the skin samples were measured. Results: A total of 60 rats were used in the study. A significant decrease in the wound size was observed in treated animals with Pimpinella anisum compared with other groups during the experiment. Additionally, treatment with Pimpinella anisum decreased the number of lymphocytes and improved the number of fibroblasts at the earlier stages and increased a number of fibrocytes at the later stages of wound healing. Other parameters such as re-epithelialisation, tissue alignment, greater maturity of collagen fibres and large capillary-sized blood vessels revealed significant changes when compared with the control. Pimpinella anisum significantly reverted oxidative changes of total antioxidant capacity, malondialdehyde and glutathione peroxidase induced by diabetic wounds (p<0.05). Furthermore, it significantly increased the dry matter and hydroxyproline contents at various stages of wound healing (p<0.05). Conclusion: The present study showed that application of Pimpinella anisum extract promotes wound healing activity in diabetic rats. The wound-healing property of Pimpinella anisum can be attributed to the phytoconstituents present in the plant

Hydrogen peroxide: a potent inducer of differentiation of human adipose-derived stem cells into chondrocytes

Common protocols for chondrogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs) are generally expensive and time-consuming and, so far, have not successfully recreated pure chondrocytes. We hypothesise that a low level of H2O2 may induce differentiation of ADSCs into chondrocytes in a shorter incubation time and relatively lower cost. Therefore, this study aimed to comparatively investigate the effectiveness of H2O2-containing or free medium in the induction of ADSCs to chondrocytes. ADSCs were isolated from the lipoaspirate of four healthy females and evaluated by immunophenotyping for their CD90, CD73, CD44, CD34, and CD45 cell surface markers. Chondrogenic differentiation was carried out using differentiation medium in the presence or absence of 10 and 50 µM H2O2 in normal and three-dimensional culture system. The intracellular contents of reactive oxygen species (ROS) were detected by flow cytometry and fluorescence microscopy. The hydroxyproline, was assessed as marker of collagen and the glycosaminoglycans (GAGs) content was both qualitatively detected and quantitatively determined. Real-time PCR was performed to determine the gene expression level of aggrecan (ACAN), type-II collagen, and transcription factor Sox9. H2O2-treated cells showed pre-chondrocyte morphology on day 1 and chondrocyte pellets were formed on day 14. H2O2-treated cells induced greater pellet sizes and showed significantly higher content of GAGs and hydroxyproline level compared with untreated cells. The gene expression levels of ACAN, collagen type-II, and Sox9 were markedly upregulated by H2O2. Our findings showed for the first time that H2O2-containing differentiation medium is potentially more effective than H2O2-free differentiation medium in the induction of chondrogensis of ADSCs. © 2018 Informa UK Limited, trading as Taylor & Francis Grou

The ability of H2O2 to induce differentiation of human mesenchymal stem cells into chondrocytes

Background and Aim: Cartilage disorders may deteriorate following oxidative stress injuries affecting mature chondrocytes. Meantime, mesenchymal stem cells (MSCs) can differentiate into chondrocytes in the presence of oxidative conditions and act as a source of compensation for injured chondrocytes. The present study aimed to investigate the effect of H2O2 on MSCs differentiation into chondrocytes in order to cast light on the dual roles of oxidative stress in the pathogenesis of diseases. Materials and Methods: Human mesenchymal stem cells were isolated from abdominal adipose tissue of three different donors and cultured in the presence of 50 μM H2O2 in order to differentiate into chondrocytes. We determined cell viability by tetrazolium assay and measured reactive oxygen species (ROS) level by flow cytometry. Presence of glycoseaminoglycans was confirmed by safranin staining. Results: The percentage of cells containing ROS was significantly higher in the cells treated with hydrogen peroxide (29.2 ± 1) compared to that in the untreated control cells (7.7 ± 1.4). A significant increase in glycoseaminoglycan content was observed in H2O2 treated cells compared to that in the control cells both on the 9th day (treated: 1.57�104 ± 0.1 vs control: 0.91�104 ± 0.09) and 21st day (treated: 2.87�104 ± 0.2 vs control: 0.96�104 ± 0.07). In addition, comparison of glycoseaminoglycan content on the 9th and 21st days showed a significantly higher content in both treated and control cells on the 21st day (p&lt;0.05). Conclusion: Hydrogen peroxide resulted in increased differentiation of adipose tissue-derived MSCs into chondrocytes. Therefore, we concluded that, oxidative stress had positive role in the induction of chondrocyte differentiation. © 2018, Kurdistan University of Medical Sciences. All rights reserved

The ability of H2O2 to induce differentiation of human mesenchymal stem cells into chondrocytes

Background and Aim: Cartilage disorders may deteriorate following oxidative stress injuries affecting mature chondrocytes. Meantime, mesenchymal stem cells (MSCs) can differentiate into chondrocytes in the presence of oxidative conditions and act as a source of compensation for injured chondrocytes. The present study aimed to investigate the effect of H2O2 on MSCs differentiation into chondrocytes in order to cast light on the dual roles of oxidative stress in the pathogenesis of diseases. Materials and Methods: Human mesenchymal stem cells were isolated from abdominal adipose tissue of three different donors and cultured in the presence of 50 μM H2O2 in order to differentiate into chondrocytes. We determined cell viability by tetrazolium assay and measured reactive oxygen species (ROS) level by flow cytometry. Presence of glycoseaminoglycans was confirmed by safranin staining. Results: The percentage of cells containing ROS was significantly higher in the cells treated with hydrogen peroxide (29.2 ± 1) compared to that in the untreated control cells (7.7 ± 1.4). A significant increase in glycoseaminoglycan content was observed in H2O2 treated cells compared to that in the control cells both on the 9th day (treated: 1.57�104 ± 0.1 vs control: 0.91�104 ± 0.09) and 21st day (treated: 2.87�104 ± 0.2 vs control: 0.96�104 ± 0.07). In addition, comparison of glycoseaminoglycan content on the 9th and 21st days showed a significantly higher content in both treated and control cells on the 21st day (p&lt;0.05). Conclusion: Hydrogen peroxide resulted in increased differentiation of adipose tissue-derived MSCs into chondrocytes. Therefore, we concluded that, oxidative stress had positive role in the induction of chondrocyte differentiation. © 2018, Kurdistan University of Medical Sciences. All rights reserved

Combined effect of retinoic acid and calcium on the in vitro differentiation of human adipose-derived stem cells to adipocytes

Context: It has been shown that adipogenesis can be modulated by factors such as all-trans retinoic acid (ATRA) and calcium. Objective: To determine, the combined effect of ATRA and calcium on the differentiation of human adipose-derived stem cells (hADSCs). Methods: Mesenchymal stem cells (MSCs) were differentiated into the adipocytes by 0.5 and 1 µM of ATRA and 5 and 10 mM calcium separately or in combination. After MTS assay the differentiation of MSCs to adipocyte was evaluated, Oil Red O staining, GLUT4 concentration and gene expression of PPARG2, adiponectin, and GLUT4 were measured by Real-Time PCR. Results: Except 10 mM calcium treated group, other groups and more significantly combination treatments could reduce all adipocyte markers compared to the control. Conclusion: These results suggest that ATRA and calcium together have significant inhibitory effect on adipogenesis that can be helpful for finding new mechanisms to prevent or control the adipogenesis. © 2017 Informa UK Limited, trading as Taylor & Francis Group

Protective effects of combined Losartan and Nilotinib on carbon tetrachloride (CCl4)-induced liver fibrosis in rats

Tyrosine kinase inhibitors (TKIs) have been developed as therapeutic compounds for inhibiting the progression of liver fibrosis. In the present study, the simultaneous treatment of Nilotinib (TKIs) and Losartan was studied. Forty rats were divided into eight groups of fibrosis induced by carbon tetrachloride (CCl4) and therapeutics (Nilotinib, Losartan, and combination therapy). In the end, serum parameters of the liver and gene expression analysis of transforming growth factor-β1, its receptors (TβRII), platelet-derived growth factor, its receptors (PDGFRβ), matrix metalloproteinases (MMP-2 and MMP-9), tumor necrosis factor-α, cytochrome P450 2E1, and collagen1 type 1 were performed. The oxidant/antioxidant factors were also analyzed. Histopathology analysis along with α-SMA immunohistochemistry and hydroxyproline evaluation was also conducted for a more in-depth study. The overall results indicated a better therapeutic effect of co-treatment of Nilotinib–Losartan in comparison with the treatment of each of them alone. Interestingly, some gene and protein factors and fibrotic indices were reduced even to the normal levels of the control group. The results of this study suggest that co-administration of these two combinations, strengthens their anti-fibrotic properties and, due to the routine use of these compounds against AML and blood pressure, these compounds can be used with caution against human liver fibrosis. © 2018, © 2018 Informa UK Limited, trading as Taylor & Francis Group

Protective effects of combined Losartan and Nilotinib on carbon tetrachloride (CCl 4 )-induced liver fibrosis in rats

Tyrosine kinase inhibitors (TKIs) have been developed as therapeutic compounds for inhibiting the progression of liver fibrosis. In the present study, the simultaneous treatment of Nilotinib (TKIs) and Losartan was studied. Forty rats were divided into eight groups of fibrosis induced by carbon tetrachloride (CCl 4 ) and therapeutics (Nilotinib, Losartan, and combination therapy). In the end, serum parameters of the liver and gene expression analysis of transforming growth factor-β 1 , its receptors (TβRII), platelet-derived growth factor, its receptors (PDGFR β ), matrix metalloproteinases (MMP-2 and MMP-9), tumor necrosis factor-α, cytochrome P450 2E1, and collagen1 type 1 were performed. The oxidant/antioxidant factors were also analyzed. Histopathology analysis along with α-SMA immunohistochemistry and hydroxyproline evaluation was also conducted for a more in-depth study. The overall results indicated a better therapeutic effect of co-treatment of Nilotinib�Losartan in comparison with the treatment of each of them alone. Interestingly, some gene and protein factors and fibrotic indices were reduced even to the normal levels of the control group. The results of this study suggest that co-administration of these two combinations, strengthens their anti-fibrotic properties and, due to the routine use of these compounds against AML and blood pressure, these compounds can be used with caution against human liver fibrosis. © 2018, © 2018 Informa UK Limited, trading as Taylor & Francis Group