Aim To evaluate the effect of MTA and Biodentine on viability, osteogenic differentiation and BMP-2 expression in osteogenic cells. Methodology Saos-2 cells were used as a model of osteoblastic cells. Overexpression of BMP-2 was induced by transfection of a CMV-driven plasmid construct including the human BMP-2 coding sequence, and stably transfected cells were selected. Cell viability was assessed by the mitochondrial dehydrogenase enzymatic (MTT) assay. The bioactivity of the materials was evaluated by the alkaline phosphatase (ALP) assay and detection of calcium deposits with alizarin red staining (ARS). The gene expression of BMP-2 and ALP was quantified with real-time PCR. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (a = 0.05). Results Viability tests revealed that MTA and Biodentine were not cytotoxic at the higher dilution (1 : 8) to BMP-2-transfected cells. MTA and Biodentine exhibited the highest ALP activity when compared to the Saos-BMP-2-unexposed control group (P < 0.05). Cell exposure to Biodentine and MTA had a significant stimulatory effect on the formation of mineralized nodules (P < 0.05). The highest increase in BMP-2 gene expression was observed after 3 days of BMP-2-transfected cells exposure to MTA and Biodentine in non-osteogenic medium in relation to Saos-BMP-2-unexposed control cells (P < 0.05). Exposure of cells to MTA in osteogenic medium for 1 day increased ALP gene expression by approximately 1.3-fold in relation to Saos-BMP-2-unexposed control cells (P < 0.05). Conclusions Both MTA and Biodentine showed biocompatibility and bioactivity in Saos-BMP-2 overexpressing cells. Biodentine had a significantly greater effect on mineralization than MTA. Both MTA and Biodentine enhanced BMP-2 mRNA expression in the transfected system. Both MTA and Biodentine are suitable materials to improve osteoblastic cell mineralization.
The aim of the present study is to evaluate the effect of hot water extract of black tea in regenerating beta cells in streptozotocin-induced diabetic mice. Light microscopic examination of pancreatic sections of streptozotocin-induced diabetic mice showed the acinar cells to be small, shrunken, and with deteriorated beta cells. The dose of streptozotocin not only altered the function of beta cells but also damaged the acinar region. The changes in acinar cells were coarsening of endoplasmic reticulation suggesting alteration in their secretory function. The control pancreatic tissue showed well-defined granulated islets and dark beta cells when stained with chrome hematoxylin and phloxine. Interestingly, pancreatic sections of diabetic mice fed with black-tea extract showed regeneration of beta cells and acinar region appeared normal with increased numbers of beta cells. To understand the probable mechanism of action of black-tea extract, we analyzed inducible nitric oxide synthase (iNOS) expression by immunohistochemistry and the results showed an increased iNOS levels in streptozotocin-induced diabetic pancreas, and such high iNOS levels were inhibited in black-tea extract treated mice. According to histological results obtained, it can be concluded that the black-tea extract helps in regeneration of damaged pancreas and protects pancreatic beta cells by its antioxidant action against nitrosative stress in streptozotocin-induced diabetes.
Hemocyanins are large extracellular respiratory proteins distributed within the hemolymph of arthropods, molluscs and larval stages of certain insects. In the present study, we characterized the humoral immune functions of cation specific antibacterial hemocyanin (Ab-Hcy) from mud crab, S. serrata. The bacteriolytic activity of Ab-Hcy was against pathogenic and non-pathogenic to crustaceans which includes, Bacillus sp. N1, B. flexus N3, E. coli, P. aeruginosa, V. harveyi, V. parahaemolyticus and V. vulnificus and also expressed bacteriostatic activity against E. coli, B. flexus N3 and V. harveyi. The agglutination activity of Ab-Hcy ranged from 4-16 against B. flexus N3, E. coli, V. harveyi and V. vulnificus and Ab-Hcy also agglutinated with rat and human O erythrocytes (HA titer: 2). The Ab-Hcy lysed the panel of mammalian erythrocytes and strong hemolytic activity of 1.40, 1.28 and 1.22 units•min −1 •mg protein −1 against human A, B and O erythrocytes respectively. It was also found that the Ab-Hcy possess phenoloxidase (PO) activity by oxidizing L-DOPA. These results suggest that Ab-Hcy from the serum of mud crab, S. serrata, is capable of performing multiple humoral immune functions in addition to oxygen transportation. Statement of relevance: This manuscript deals with the humoral immune functions, particularly antibacterial property of hemocyanin in crustacean. In this paper we report the different type of antibacterial activity and other humoral immune functions of a hemocyanin isolated from the serum of mud crab, Scylla serrata. This work will help to understand the role of respiratory molecule in immune system, also will be helpful improve disease free crustacean aquaculture industry. And we also state this research work is relevant and suitable to publish in the Journal "Aquaculture".
The aim of this study is to investigate the mechanism of an action of compound isolated from Vitex negundo in streptozotocin-induced diabetic mice. Light microscopic examination of liver, kidney and pancreatic sections of streptozotocin-induced diabetic mice showed changes like coarsening of acinar cells of endoplasmic reticulum, destruction of β-cells, and alteration in their secretory function were observed in the pancreas. Changes like dilation of vein, unusual concentric arrangement of hepatocytes, and liver fibrosis were observed in the liver. Thickening of tubules and expansion of glomerulus were observed in kidneys. All these altered parameters were reversed close to normal condition upon treatment using idopyranose. The results show the antidiabetic potential of idopyranose. Interestingly, liver, kidney, and pancreatic sections of diabetic mice fed with the isolated 1, 2 di-substituted idopyranose showed regeneration of hepatocytes, nephrocytes, as well as β-cells and acinar region appeared normal with increased numbers of β-cells. To understand the probable mechanism of action of 1, 2 di-substituted idopyranose, we analyzed proinflammatory inducible nitric oxide synthase (iNOS) and nuclear factor-kappa B (NF-κB) expression by immunohistochemistry and the results showed an increased iNOS and NF-κB levels in streptozotocin-induced diabetic liver, kidney and pancreas. Such high iNOS and NF-κB levels were inhibited in 1, 2 di-substituted idopyranose treated mice. The results suggest that 1, 2 di-substituted idopyranose helps in the protection of hepatocytes, nephrocytes and pancreatic β-cells probably by its action against NF-κB and iNOS mediated inflammation in streptozotocin-induced diabetes.
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