Mtt Assay
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2022 ◽  
Vol 12 (5) ◽  
pp. 920-925
Author(s):  
He Bai ◽  
Jian He

The BMSCs are one of the components of tumor micro-environment and participate in tumor evolution. Our study aimed to discuss the effect of exosome derived from BMSC on gastric cancer cells. Tumor and para-tumor tissues were isolated to measure miR-206 level by RT-PCR. Gastric cancer cell behaviors were analyzed using MTT assay and scratch test. Gastric cancer model was established and treated TIGIT inhibitor to assess its role in the tumor growth in vivo. The miR-206 in exosome from BMSCs in cancer tissue was detected. CD8 expression excreted by DC could be induced after miR-206 treatment possibly through regulating the signaling pathway of TIGIT/PVR. Inhibition of TIGIT decreased tumor growth, development and reversed tumor phenotype. In conclusion, miR-206 derived from BMSCs induces CD8 expression in gastric cancer through regulating the signaling pathway of TIGIT/PVR, indicating that it might be a novel target for the treatment of gastric cancer.


2022 ◽  
Vol 12 (5) ◽  
pp. 971-977
Author(s):  
Ruoyu Zhu ◽  
Zhonglin Wang

This study investigated the impact of microRNA (miR)-376b derived from BMSCs on glioma progression. BMSCs were transfected with miR-376b mimic, miR-376b inhibitor or NC and then cocultured with glioma cells followed by measuring cell behaviors by MTT assay, Transwell assay and flow cytometry, FOXP2 and miR-376b expression by Western blot and RT-qPCR. After confirming the inhibitory and mimicking activity of transfection, we found that overexpression of miR-376b in BMSCs decreased glioma cell invasion, migration and proliferation but promoted cell apoptosis within 24 h and 48 h after transfection along with reduced number of cells in S-phase. Mechanically, miR-376b targeted miR-376b and up-regulation of miR-376b caused down-regulation of FOXP2 (p < 0.05). Overexpression of miR-376b in BMSCs decelerated glioma cell cycle and inhibitedmalignant behaviors of glioma cells by targeting FOXP2 expression. These evidence unveils the potential role of FOXP2 as a biomarker for the treatment of gliomas.


2022 ◽  
Vol 8 (1) ◽  
pp. 185-191
Author(s):  
Mahesh Kumar D

Background: Silver Nanoparticles are extensively studied by the scientific community for therapeutic applications. With respect to the fundamental pillars of bioethics “Primum non nocere” equal emphasis should be given to evaluate the toxicological perspectives of Silver nanoparticles. This study aims at evaluating the InVitro cytotoxic effects of Silver nanoparticles synthesized using hesperidin. Aim: To study the In Vitro cytotoxicity of silver nanoparticles on PBMC cells using (3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Methods: Synthesized silver nanoparticles at various concentrations are incubated with peripheral blood mononuclear cells (PBMC). After 24 hours MTT is added to the mixture to evaluate the cell viability post incubation. Yellow MTT (a tetrazole) which is reduced to purple formazan in the mitochondria of living cells. The absorbance of this colored solution can be quantified by measuring at 570 nm by a spectrophotometer. This reduction takes place only when mitochondrial reductase enzymes are active, and therefore conversion can be directly related to the number of viable (living) cells. Results: ?.Conclusion: Silver Nanoparticles do not exhibit any significant cytotoxicity on PBMCs and also there were no dose dependent trends in the results.


Molecules ◽  
2022 ◽  
Vol 27 (2) ◽  
pp. 451
Author(s):  
Hermizi Hapidin ◽  
Nor Munira Hashim ◽  
Mohamad Zahid Kasiram ◽  
Hasmah Abdullah

Background: This study investigates the effect of tannic acid (TA) combined with pamidronate (PAM) on a human osteoblast cell line. Methods: EC50 for TA, PAM, and different combination ratios of TA and PAM (25:75, 50:50, 75:25) were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The combination index value was utilized to analyze the degree of drug interaction, while trypan blue assay was applied to analyze the cells proliferation effect. The mineralization and detection of bone BSP and Osx genes were determined via histochemical staining and PCR test, respectively. Results: The EC50 of osteoblasts treated with TA and a 75:25 ratio of TA and PAM were more potent with lower EC50 at 0.56 µg/mL and 0.48 µg/mL, respectively. The combination of TA and PAM (75:25) was shown to have synergistic interaction. On Day 7, both TA and PAM groups showed significantly increased proliferation compared with control and combination groups. On Day 7, both the TA and combination-treated groups demonstrated a higher production of calcium deposits than the control and PAM-treated groups. Moreover, on Day 7, the combination-treated group showed a significantly higher expression of BSP and Osx genes than both the TA and PAM groups. Conclusion: Combination treatment of TA and PAM at 75:25 ameliorated the highest enhancement of osteoblast proliferation and mineralization as well as caused a high expression of BSP and Osx genes.


Author(s):  
Mahdis Parsafar ◽  
Sayed Khatiboleslam Sadrnezhaad ◽  
Nahid Hassanzadeh Nemati

: Due to the increasing number of diseases related to the spine, we expect to see more research on the implants used in this area. These implants should have features such as strength, fatigue resistance, chemical stability and biocompatibility, which fortunately are seen in an alloy called nitinol. In this study, six lumbar vertebral implants were made of nitinol and these samples been studied by some experiments such as: X-ray diffraction and biocompatibility (evaluation of cytotoxicity by MTT assay). Finally, we came to the conclusion that the mentioned alloy with suitable microstructure is appropriate for medical applications specially as an orthopedic implant. According to the X-ray diffraction pattern, the samples have austenitic structures in the room temperature and the predominant phase of the porous sample is B2-NiTi. Since the sample should be biocompatible after placement in the body and should not cause an immune system reaction, this test was also examined and the samples were tested in vitro with an MTT kit and the biocompatibility was assessed. The results of biocompatibility tests also indicate the suitability of the implant in terms of cellular characteristics. These properties have made this alloy superior to other alloys in orthopedic implant utilization, especially in areas under continuous loading. It is hoped that the construction of this type of implant will pave the way for facilitating the treatment of spinal abnormalities.


2022 ◽  
Vol 25 ◽  
pp. 69-76
Author(s):  
Leyla Anari ◽  
Davood Mehrabani ◽  
Mahboobeh Nasiri ◽  
Shahrokh Zare ◽  
Iman Jamhiri ◽  
...  

Purpose: Among abused substances, methamphetamine is a psychostimulant drug widely used recreationally with public health importance. This study investigated the effect of methamphetamine on proliferation, differentiation, and apoptosis of human adipose tissue stem cells (AdSCs). Methods: AdSCs were isolated from human abdominal adipose tissue and were characterized for mesenchymal properties and growth kinetics. MTT assay was undertaken to assess methamphetamine toxicity on proliferation and differentiation properties and apoptosis of hAdSCs. Results: Isolated cells were shown to have mesenchymal properties and a population doubling time (PDT) of 40.1 h. Following methamphetamine treatment, expressions of KI-67 and TPX2 as proliferation genes and Col1A1 and PPARg as differentiation genes decreased. Methamphetamine administration increased the expression of Bax and decreased Bcl-2 genes responsible for apoptosis. Conclusions: Our data suggested when AdSCs were exposed to methamphetamine, it decreased proliferation and differentiation properties of stem cells together with an increase in apoptosis. These findings can be added to the literature, especially when methamphetamine is used recreationally for weight loss purposes.


Author(s):  
Marjan Bahraminasab ◽  
Samaneh Arab ◽  
Nesa Doostmohammadi

The aim of this study was to evaluate the biocompatibility of Al2O3-Ti functionally graded material (FGM) successfully fabricated by Spark Plasma Sintering (SPS) technology, and to compare with pure Ti and alumina. Pre-osteoblast MC3T3-E1 cells were used to examine cell viability, proliferation and differentiation using lactate dehydrogenase (LDH) cytotoxicity detection kit, MTT assay and Alkaline Phosphatase (ALP) colorimetric test at different time points. Furthermore, ion release from the materials into the culture medium was assessed. The results showed cell viability over 80% for FGM and alumina which dismissed any cytotoxicity risk due to materials or manufacturing. The results of MTT tests identified superiority of FGM than Ti and alumina, particularly in late proliferation. Nevertheless, in cell differentiation, all materials performed similarly with no statistical differences. Furthermore, it was indicated that Ti had no ion release, while alumina had small amount of Al ion dissolution. FGM, however, had more ions detachment, particularly Al ions.


Author(s):  
Alaa A. Mohammed ◽  
Jawad K. Oleiwi

Polyetheretherketone is a semi-crystalline thermoplastic polymer, that so with heat treatments, it is possible to get different properties which are very important for the material performance. Heat treatment is a broadly utilized to develop the semi-crystalline polymers properties. In the present investigation, annealing of polyetheretherketone (PEEK) was carried out at temperatures above its glass transition temperature (Tg) to study its effects upon the biological conduct of the control and PEEK ternary composites. The bioactivity of the specimens was evaluated by investigating the apatite formation after immersion for different periods in a simulated body fluid (SBF). The biocompatibility of specimens was assessed by MTT assay. Additionally, the antibacterial property of the specimens versus S. aureus was observed with the optical density methods. The results manifested that the formation of hydroxyapatite was obviously observed on specimens after immersion for (7 and 14 days) in the simulated body fluid (SBF). Otherwise, the results of MTT assay recorded the PEEK specimens that excited the activity of fibroblasts, and therefore a high cytocompatibility was noticed and the specimens revealed antibacterial properties against S. aureus. So, the results of the bioactivity, biocompatibility and antibacterial tests in vitro demonstrated that the heat treatment enhanced biological behavior.


2022 ◽  
Author(s):  
Pegah Sarraf ◽  
Razieh Sadat Moayeri ◽  
Noushin Shokouhinejad ◽  
Mehrfam Khoshkhounejad ◽  
Roya Karimi ◽  
...  

Abstract Background: PRF as one of the favorable scaffolds in Regenerative Endodontic Treatment (RET), has several limitations such as the need for blood sampling and special equipment. High available commercial scaffolds such as fibrin are able to meet all the necessary requirements of dentin tissue engineering. The present study was designed to evaluate the effect of PRF and fibrin gel, with and without the presence of EDTA-treated radicular dentin segments on SCAP viability, proliferation, migration, and differentiation.Methods: Radicular dentin were prepared from extracted teeth and treated by EDTA 17% .The samples were divided into 6 groups: Dentin/PRF/Cell, Dentin/Fibrin/Cell, Dentin/Cell, PRF/Cell, Fibrin/Cell and Cell (Control). SCAP viability was assessed using MTT assay. Gene expression levels of odontogenic markers [Dentin sialophosphoprotein (DSPP), Dentin matrix protein 1(DMP1), Collagen type I Alpha 1(COL 1A1) and Alkaline phosphatase (ALP) were assessed using qrt-PCR. Cell migration were also evaluated by means of scratch test. Results: The results of MTT assay at showed that the viability of SCAP significantly increased after 7 days for both groups containing fibrin (P <0.05). The viability of SCAP seeded on Dentin/PRF and PRF significantly decreased after 7 days (P <0.001). The odontogenic markers were significantly expressed for both scaffolds in the presence of dentin segment (p<0.05). Significant decrease in scratch area was seen in Fibrin/Dentin group (p < 0.001)Conclusions:Fibrin beside EDTA-treated dentin showed great ability in survival, proliferation, differentiation, and migration of SCAP rather than PRF.


Author(s):  
Houri Sadri ◽  
Mahmoud Aghaei ◽  
Vajihe Akbari

Nisin, an antimicrobial peptide produced by Lactococcus lactis, is widely used as a safe food preservative and has been recently attracting the attention of many researchers as a potential anticancer agent. The cytotoxicity of nisin against HeLa, OVCAR-3, SK-OV-3, and HUVEC cells was evaluated using MTT assay. The apoptotic effect of nisin was identified by Annexin-V/propidium iodide assay, and then it was further confirmed by western blotting analysis, mitochondrial membrane potential (ΔΨm) analysis, and reactive oxygen species (ROS) assay. The MTT assay showed concentration-dependent cytotoxicity of nisin towards cancer cell lines, with the IC50 values of 11.5-23 µM, but less toxicity against normal endothelial cells. Furthermore, treatment of cervical cancer cells with 12 µM nisin significantly (P<0.05) increased the Bax/Bcl-2 ratio (4.9-fold), reduced ΔΨm (70%), and elevated ROS levels (1.7-fold). These findings indicated that nisin might have anticancer and apoptogenic activities through mitochondrial dysfunction and oxidative stress damage in cervical cancer cells.


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