A novel method for evaluating the dynamic biocompatibility of degradable biomaterials based on real-time cell analysis

Abstract Degradable biomaterials have emerged as a promising type of medical materials because of their unique advantages of biocompatibility, biodegradability and biosafety. Owing to their bioabsorbable and biocompatible properties, magnesium-based biomaterials are considered as ideal degradable medical implants. However, the rapid corrosion of magnesium-based materials not only limits their clinical application but also necessitates a more specific biological evaluation system and biosafety standard. In this study, extracts of pure Mg and its calcium alloy were prepared using different media based on ISO 10993:12; the Mg2+ concentration and osmolality of each extract were measured. The biocompatibility was investigated using the MTT assay and xCELLigence real-time cell analysis (RTCA). Cytotoxicity tests were conducted with L929, MG-63 and human umbilical vein endothelial cell lines. The results of the RTCA highly matched with those of the MTT assay and revealed the different dynamic modes of the cytotoxic process, which are related to the differences in the tested cell lines, Mg-based materials and dilution rates of extracts. This study provides an insight on the biocompatibility of biodegradable materials from the perspective of cytotoxic dynamics and suggests the applicability of RTCA for the cytotoxic evaluation of degradable biomaterials.

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Decorosides A and B, Cytotoxic Flavonoid Glycosides from the Leaves of Rhododendron decorum

Natural Product Communications ◽

10.1177/1934578x1400900410 ◽

2014 ◽

Vol 9 (4) ◽

pp. 1934578X1400900 ◽

Cited By ~ 1

Author(s):

Mostafa E. Rateb ◽

Hossam M. Hassan ◽

El-Shaimaa A. Arafa ◽

Marcel Jaspars ◽

Rainer Ebel

Keyword(s):

Spectral Data ◽

Cell Lines ◽

Cancer Cell ◽

Mtt Assay ◽

Methanolic Extract ◽

Cancer Cell Lines ◽

Biological Evaluation ◽

Flavonoid Glycosides ◽

Cytotoxic Activities ◽

Diphenyltetrazolium Bromide

Bioassay and NMR-guided fractionation of the methanolic extract of Rhododendron decorum leaves resulted in the isolation of two new flavonoid glycosides, 5,7-dihydroxy-6,8-dimethyldihydroflavanone-7-O-α-L-arabinopyranosyl(l→6)-β-D-glucopyranoside (decoroside A, 1) and its 3-hydroxy congener (decoroside B, 2), along with five known compounds myricitrin (3), afzelin (4), (-)-epicatechin (5), (+)-catechin (6), and ampeloptin (7). The structures of the isolated compounds were elucidated by extensive interpretation of their spectral data. Biological evaluation using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed promising cytotoxic activities of these compounds against different cancer cell lines.

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iCELLigence real-time cell analysis system for examining the cytotoxicity of drugs to cancer cell lines

Experimental and Therapeutic Medicine ◽

10.3892/etm.2017.4781 ◽

2017 ◽

Vol 14 (3) ◽

pp. 1866-1870 ◽

Cited By ~ 17

Author(s):

Leyla Türker Şener ◽

Gürcan Albeni̇z ◽

Bi̇rcan Di̇nç ◽

Işil Albeni̇z

Keyword(s):

Real Time ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Cell Analysis ◽

Analysis System

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The role of methylation leading to capecitabine resistant malignant mesothelioma

Journal of Clinical Oncology ◽

10.1200/jco.2006.24.18_suppl.17105 ◽

2006 ◽

Vol 24 (18_suppl) ◽

pp. 17105-17105 ◽

Cited By ~ 2

Author(s):

K. V. Kosuri ◽

X. Wu ◽

G. Otterson

Keyword(s):

Real Time ◽

Cell Lines ◽

Malignant Mesothelioma ◽

Mtt Assay ◽

Real Time Pcr ◽

Dihydropyrimidine Dehydrogenase ◽

Relative Difference ◽

Western Blots ◽

Mesothelioma Cell

17105 Background: Malignant mesothelioma is a deadly malignancy whose global incidence continues to be on the rise. Established therapies have been less than optimal. The current therapeutic standard is intravenous pemetrexed, an antifolate medication. Yet, another folate antimetabolite, capecitabine, is significantly less effective than pemetrexed. The enzymes thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), and thymidylate phosphatase (TP) are critical to the efficacy of antifolates. Specifically, for capecitabine to be converted into a potent cytotoxic agent, the enzyme TP must be present and active. In one of four mesothelioma cell lines examined, the gene that encodes for TP, extracellular growth factor-1 (ECGF-1), is methylated. Methylation of this gene and the subsequent downregualtion of the TP enzyme confer a diminished cytotoxic effect by a capecitabine prodrug, dioxyfluridine (DFUR). Methods: Cells were cultured treated with and without 1uM decitabine (DAC) under identical conditions. DNA, RNA, and protein lysates were collected after 72 hours. Bisulfite-treated DNA was examined by MS-PCR for evidence of methylation of TS, DPD, and TP. RNA was collected and cDNA was synthesized. Real time PCR was utilized to detect the relative difference in RNA quantity. Western blots were done to evaluate the differences in protein expression between DAC treated and untreated cells. MTT assay was performed with DAC pretreated and untreated cell lines subsequently treated with DFUR and 5-fluorouracil. Results: One of the four mesothelioma cell lines showed consistent evidence of TP methylation by MS-PCR. The addition of 1uM DAC to the cell lines conferred a six-fold difference in expression of the methylated gene by both real time PCR as well as by Western blot. The prodrug DFUR subsequently shows increased cytotoxicity in the methylated cell line by MTT assay when pretreated with DAC compared when not exposed to the DAC. Conclusion: By demethylating the ECGF-1 gene with DAC, the now upregulated TP enzyme has an increased ability to convert DFUR to 5-FU; thus enhancing the cytotoxicity of a drug thought to be ineffective in malignant mesothelioma. No significant financial relationships to disclose.

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Phenotypic testing of patient herpes simplex virus type 1 and 2 isolates for acyclovir resistance by a novel method based on real-time cell analysis

Journal of Clinical Virology ◽

10.1016/j.jcv.2020.104303 ◽

2020 ◽

Vol 125 ◽

pp. 104303 ◽

Cited By ~ 2

Author(s):

Oliver Caliaro ◽

Maria Teresa Barbani ◽

Shkipe Klenja ◽

Florence Morfin ◽

Emilie Frobert ◽

...

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Real Time ◽

Virus Type ◽

Herpes Simplex Virus Type ◽

Cell Analysis ◽

Novel Method ◽

Simplex Virus

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Synthesis and Biological Evaluation of New Quinoline-Based Thiazolyl Hydrazone Derivatives as Potent Antifungal and Anticancer Agents

Letters in Drug Design & Discovery ◽

10.2174/1570180814666171003145227 ◽

2018 ◽

Vol 15 (2) ◽

pp. 193-202 ◽

Cited By ~ 6

Author(s):

Ali Erguc ◽

Mehlika Dilek Altintop ◽

Ozlem Atli ◽

Belgin Sever ◽

Gokalp Iscan ◽

...

Keyword(s):

Anticancer Activity ◽

Cell Lines ◽

Mtt Assay ◽

Anticancer Agents ◽

Mouse Embryonic Fibroblast ◽

Biological Evaluation ◽

Diffusion Method ◽

Breast Adenocarcinoma ◽

Nih 3T3 ◽

Mcf 7

Background: In medicinal chemistry, thiazoles have gained great importance in antifungal and anticancer drug design and development. Objectives: The aim of this study was to synthesize new quinoline-based thiazolyl hydrazone derivatives and evaluate their anticandidal and anticancer effects. Methods: New thiazolyl hydrazone derivatives were evaluated for their anticandidal effects using disc diffusion method. Ames MPF assay was carried out to determine the genotoxicity of the most effective antifungal derivative. MTT assay was also performed to assess the cytotoxic effects of the compounds on A549 human lung adenocarcinoma, HepG2 human hepatocellular carcinoma, MCF- 7 human breast adenocarcinoma and NIH/3T3 mouse embryonic fibroblast (healthy) cell lines. Methods: Results: 4-(4-Fluorophenyl)-2-(2-((quinolin-4-yl)methylene)hydrazinyl)thiazole (4) showed antifungal activity against Candida albicans and Candida krusei in the concentration of 1 mg/mL. In MTT and Ames MPF tests, it was determined that compound 4 did not show cytotoxic and genotoxic effects. MTT assay indicated that 4-(naphthalen-2-yl)-2-(2-((quinolin-4-yl)methylene) hydrazinyl)thiazole (10) showed more selective anticancer activity than cisplatin against A549 and MCF-7 cell lines. Besides, 4-(4-chlorophenyl)-2-(2-((quinolin-4-yl)methylene)hydrazinyl)thiazole (5) exhibited more selective anticancer activity than cisplatin against HepG2 cell line. Conclusion: Due to their high selectivity index, these compounds are considered as candidate compounds to participate in further research.

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Design, Synthesis of novel coumarin conjugated acetamides as promising anticancer agents: An in-silico and in-vitro approach.

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200714140820 ◽

2020 ◽

Vol 20 ◽

Author(s):

Mamatha S. V ◽

S. L. Belagali ◽

Mahesh Bhat ◽

Vijay M. Kumbar

Keyword(s):

Anticancer Activity ◽

Cell Lines ◽

Mtt Assay ◽

Anticancer Agents ◽

In Silico ◽

Active Sites ◽

Biological Activities ◽

Biological Evaluation ◽

Mcf 7

Background: Coumarin and benzophenone possess a vast sphere of biological activities whereas thiazoles display various pharmacological properties. Hence we focused on incorporation of coumarin and thiazole core to the benzophenone skeleton to enhance the bioactivity anticipating their interesting biological properties. Objective: The objective of the current work is synthesis and biological evaluation of a novel series of coumarin fused thiazole derivatives. Methods: A novel series of Coumarin conjugated thiazolyl acetamide hybrid derivatives were synthesized by multistep reaction sequence and were characterized by the FT-IR, LCMS and NMR spectral techniques. The newly synthesized compounds were screened for anticancer activity by in-silico and in-vitro methods. The cytotoxicity of the synthesized unique compounds had been executed for two different cancer cell lines MCF-7 (Breast cancer) and KB (Oral cancer) in comparison with standard paclitaxel by MTT assay. Results: The compound 7f is the potent motif with an acceptable range of IC 50 values for anticancer activity were 63.54 µg/ml and 55.67 µg/ml, against the MCF-7 and KB cell lines, respectively. Molecule docking model revealed that this compound formed three conventional hydrogen bonds with the active sites of the amino acids MET 769, ARG 817 and LYS 721. Conclusion: Compound 7f with two methyl groups on the phenoxy ring and one 4-position methoxy group on the benzoyl ring, showed a significant cytotoxic effect. An advantageous level of low toxicity against normal cell line (L292) by MTT assay was determined.

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Cyanidin 3-O-Glucoside Induces the Apoptosis in the Osteosarcoma Cells through Upregulation of the PPARγ and P21: An In Vitro Study

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200408081111 ◽

2020 ◽

Vol 20 (9) ◽

pp. 1087-1093

Author(s):

Hesam A. Atashi ◽

Hamid Z. Arani ◽

Amirhossein Shekarriz ◽

Hamidreza Nazari ◽

Amirhossein Zabolian ◽

...

Keyword(s):

Real Time ◽

Cell Lines ◽

Mtt Assay ◽

Malignant Tumors ◽

Apoptotic Cell ◽

Annexin V ◽

In Vitro Study ◽

Osteosarcoma Cell ◽

Osteosarcoma Cells

Background: Osteosarcoma (OS) is known as the malignant tumors in the bone. Cyanidin 3-OGlucoside (C3G) has a potential to induce the apoptotic cell death in different cancer cells; however, the mechanisms of action for C3G have not been clarified yet. Objective: In this study, the apoptotic effects of C3G on three different osteosarcoma cell lines including Saso-2, MG-63, and G-292 (clone A141B1) were investigated. Methods: The 24-hr IC50 of C3G for Saso-2, G-292, and MG-63 cells was evaluated by the MTT assay. Apoptosis induction in these cell lines after treatment with the C3G was approved by the Annexin V/PI flow cytometry. Changes at the mRNA expression level of PPARγ, P21, Bax, and Bcl-xl genes were investigated by real-time Polymerase Chain Reaction (PCR) technique, and P21 expression was further confirmed by the western blotting. Results: The MTT assay results demonstrated that the 24-hr IC50 of C3G was equal to 110μg/ml for Saso-2 and G-292 cells while it was about 140μg/ml for the MG-63 cells. The results of real-time PCR clearly showed that treatment of the cells with 24hrs IC50 of C3G caused the upregulation of PPARγ, P21, and Bax genes. Moreover, western blot analysis confirmed that P21 protein overexpressed endogenously after treatment of the cells with the C3G, and it was more upregulated in the MG-63 cells compared to the other cell lines. Conclusion: According to the findings of the study, the C3G is a novel anti-osteosarcoma agent with the ability to induce the apoptosis in different osteosarcoma cells through upregulation of the PPARγ and P21 genes.

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