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Alternative Methods for Assessing the Effects of Chemicals in the Eye
Published in David W. Hobson, Dermal and Ocular Toxicology, 2020
Leon H. Bruner, John Shadduck, Diane Essex-Sorlie
Selected technique — Highest tolerated dose.37,39–41 BALB/c 3T3 cells are seeded into 96-well culture plates at densities sufficient to obtain semiconfluent cultures 24 h after seeding. Growth medium is removed and replaced with medium containing the test chemicals; then, the cells are incubated in the presence of the chemical for 24 h. Cells are observed microscopically and evaluated for morphologic alterations using phase contrast microscopy. Various concentrations of the test chemical are used, and the highest concentration of the chemical that does not produce morphological alterations is designated the highest tolerated dose. Unexposed BALB/c 3T3 cells are used for comparison. The indices evaluated include decreased cell density, increased cell roundedness, and cellular cytoplasmic granularity. In terms of ranking a chemical relative to others in a set, agreement with other in vitro tests and rabbit eye irritation test data has been good in a limited number of evaluations.
Mathematical Models of Irradiated Cell Survival
Published in Leonid G. Hanin, Lyudmila V. Pavlova, Andrej Yu. Yakovlev, Biomathematical Problems in Optimization of Cancer Radiotherapy, 2020
Leonid G. Hanin, Lyudmila V. Pavlova, Andrej Yu. Yakovlev
No significant difference between stationary and exponential states of growth was found for HeLa cells as far as numerical parameters of model 5.1 were concerned. It is not that surprising in view of the fact that, being irradiated in different phases of their growth, HeLa cells demonstrate virtually identical dose-effect patterns (Figure 2). On the contrary, marked differences manifest themselves for the two states of cell culture when results with 3T3 cells are examined (see Table 4 and Figure 3). In the stationary phase, 3T3 cells are considerably less sensitive than in the exponential phase as is reflected by the lower value of the parameter μ. At the same time, the critical number of radiation-induced lesions also appears to be lower for these cells in the stationary phase of their growth than in the exponential phase. This discrepancy can possibly be explained by a more efficient sublethal damage repair in the exponentially growing cells. Thus, the actively proliferating 3T3 cells simultaneously possess both a higher sensitivity to radiation (the mean number of the radiation-induced lesions per unit dose) and a greater capacity for sublethal damage repair (the critical number of hits) than their resting counterparts.
Cellular and Viral Oncogenes
Published in Pimentel Enrique, Oncogenes, 2020
The cells used as recipients in most transfection experiments are NIH/3T3 cells, which are fibroblasts of murine (mouse) origin maintained as a contact-inhibited, nontumorigenic cell line. These cells have been considered as normal cells because they grow in sheets with the characteristic flattened appearance of fibroblasts, forming a confluent monolayer in Petri dishes, after which there is no more growth. However, as other immortal cell lines, NIH/3T3 cells cannot strictly be considered as normal cells. They acquire clearly evident neoplastic characteristics a few weeks after being transfected with high molecular weight (>30 kb)
Is a non-cytotoxic and non-genotoxic novel bioinspired dipeptide structure synthesis possible for theragnostic applications?
Published in Drug and Chemical Toxicology, 2023
Merve Bacanlı, Jülide Secerli, Burcu Karayavuz, Onur Erdem, Hakan Erdoğan
NIH/3T3 cells were cultured as described in 2.4. 3–(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed by the method of Mosmann (1983) with the modifications of Kuźma et al. (2012). A total of 105 cells/well were plated in 96 well tissue-culture plates and the cells were incubated for 24 h at 37 °C in an atmosphere supplemented with 5% CO2. After the cells adhere to the plates, various concentrations (1, 5, 10, and 20 µg/mL) of Hg2+/Phe–Phe dipeptides for 24, 48, and 72 h in full medium were applied to the cells. Full medium was used as negative control. After the exposure, the medium was aspirated, cells were washed with PBS and MTT solution (5 mg/mL of stock in PBS) was added (10 µL/well in 100 µL of cell suspension), and cells were incubated for an additional 3-h with MTT. At the end of incubation period, the dye was carefully removed and 100 µL of DMSO was added to each well. The absorbance of the solution was measured in a microplate reader (Reader Epoch, Biotek, Winooski, VT) at 570 nm.
Anticarcinogenic potential of gold nanoparticles synthesized from Trichosanthes kirilowii in colon cancer cells through the induction of apoptotic pathway
Published in Artificial Cells, Nanomedicine, and Biotechnology, 2019
Xiaodong Han, Xiaojia Jiang, Lanjie Guo, Yongxin Wang, Vishnu Priya Veeraraghavan, Surapaneni Krishna Mohan, Zhigang Wang, Dandan Cao
The anti-carcinogenic effect of phytoconstiuent-mediated AuNPs creates a center of attention on growing interest in AuNPs synthesis, which are being well thought-out as novel agents. In addition, AuNPs act as nanocarriers in the targeted system of drug delivery [29]. In this report, we elucidated the therapeutic potential anti-carcinogenic effect of AuNPs on HCT-116 cancer cells and 3T3 cells. In order to find out the safe working concentration, HCT-116 cells were treated with 5, 10, 15, 20, and 25 μg/ml concentrations of AuNPs. These findings showed AuNPs considerably reduced the cell viability by 86.47%, 71.25%, 61.14%, 53.24%, 41.29% and 29.87% at 5, 10, 15, 20, and 25 μg/ml concentration, respectively; approximately 50% cell viability inhibition was documented with AuNPs at 15.5 μg/ml (Figure 4(A)). Particularly, HCT-116 cells were very much sensitive to T. kirilowii extract mediated AuNPs based on a concentration-dependent manner. However, no significant toxicity was observed (Figure 4(B)) in normal 3T3 cells which clearly shows the safety nature to the normal cells.
A novel conjunction of folate-targeted carbon nanotubes containing protohemin and oridonin-liposome loaded microbubbles for cancer chemo-sonodynamic therapy
Published in Journal of Drug Targeting, 2019
Chuan-Jin Wang, Heng-Zhi Wang, Wei Li
The cytotoxicity induced by blank LUM, ultrasound alone, FMT alone, Ph alone, MTP alone, FMTP alone, ORI alone, LO alone, LUMO alone, MTP-LUMO alone, FMTP-LUMO alone, Blank LUM plus ultrasound, FMT plus ultrasound, Ph plus ultrasound, MTP plus ultrasound, FMTP plus ultrasound, ORI plus ultrasound, LO plus ultrasound, LUMO plus ultrasound, MTP-LUMO plus ultrasound, FMTP-LUMO plus ultrasound on HepG-2 cells were tested by the CCK-8 method, as reported previously [28,29]. Briefly speaking, 3T3 cells and HepG-2 cells were firstly counted, then transferred to 96-well microtiter plates, and sequentially incubated for 24 h previous to the addition of test drug carriers. Compounds were dispersed in DMSO and diluted with sterile water for injection to obtain the suitable concentration (ORI concentration of 12 μg/mL and/or Ph concentration of 24 μg/mL). Inhibition of 3T3 cells and HepG-2 cell growth of chemo-sonodynamic therapy were investigated according to our previous work [27].