Introducing Wound Healing Assays in the Undergraduate Biology Laboratory Using Ibidi Plates

Suh, Yewseok; Robinson, A; Zanghi, Nicholas; Kratz, Austin; Gustetic, Andrew; Crow, Mackenzie M.; Ritts, Taylor; Hankey, William; Segarra, Verónica A.

doi:10.1128/jmbe.00061-22

Cited by 5 publications

(4 citation statements)

References 7 publications

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“…The sample treatment includes control, 5 mmol/L Met, and 1,000 V/cm + 5 mmol/L Met. The culture-insert 2-well from Ibidi ® was used to create a defined cell-free gap of 500 µm in width [ 83 ]. The 10 mm culture dish was sterilized, and the culture-insert 2-well was inserted, creating two individual wells separated by a central divider.…”

Section: Methodsmentioning

confidence: 99%

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Efficacy of metformin and electrical pulses in breast cancer MDA-MB-231 cells

Sahu,

Camarillo,

Sundararajan

2024

Exploration of Targeted Anti-Tumor Therapy

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Aim: Triple-negative breast cancer (TNBC) is a very aggressive subset of breast cancer, with limited treatment options, due to the lack of three commonly targeted receptors, which merits the need for novel treatments for TNBC. Towards this need, the use of metformin (Met), the most widely used type-2 diabetes drug worldwide, was explored as a repurposed anticancer agent. Cancer being a metabolic disease, the modulation of two crucial metabolites, glucose, and reactive oxygen species (ROS), is studied in MDA-MB-231 TNBC cells, using Met in the presence of electrical pulses (EP) to enhance the drug efficacy. Methods: MDA-MB-231, human TNBC cells were treated with Met in the presence of EP, with various concentrations Met of 1 mmol/L, 2.5 mmol/L, 5 mmol/L, and 10 mmol/L. EP of 500 V/cm, 800 V/cm, and 1,000 V/cm (with a pulse width of 100 µs at 1 s intervals) were applied to TNBC and the impact of these two treatments was studied. Various assays, including cell viability, microscopic inspection, glucose, ROS, and wound healing assay, were performed to characterize the response of the cells to the combination treatment. Results: Combining 1,000 V/cm with 5 mmol/L Met yielded cell viability as low as 42.6% at 24 h. The glucose level was reduced by 5.60-fold and the ROS levels were increased by 9.56-fold compared to the control, leading to apoptotic cell death. Conclusions: The results indicate the enhanced anticancer effect of Met in the presence of electric pulses. The cell growth is inhibited by suppressing glucose levels and elevated ROS. This shows a synergistic interplay between electroporation, Met, glucose, and ROS metabolic alterations. The results show promises for combinational therapy in TNBC patients.

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Section: Methodsmentioning

confidence: 99%

“…A wound healing assay was used to study the cell migration [82] of MDA-MB-231 cells under the influence of Met and electric pulse. The sample treatment includes control, 5 mmol/L Met, and 1,000 V/cm + 5 mmol/L Met.…”

Section: Wound Healing Assaymentioning

confidence: 99%

Efficacy of metformin and electrical pulses in breast cancer MDA-MB-231 cells

Sahu,

Camarillo,

Sundararajan

2024

Exploration of Targeted Anti-Tumor Therapy

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“…inhibit hyperphosphorylated tau via the upregulation of IRS-1/PI3K/Akt and downregulation of GSK-3β. Molecular docking results further demonstrate that these polyphenols exhibit a good binding property with insulin receptors [ 135 ].…”

Section: Polyphenols Vs Neurodegenerative Diseasementioning

confidence: 99%

Neuro-Nutraceutical Polyphenols: How Far Are We?

et al. 2023

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The brain, composed of billions of neurons, is a complex network of interacting dynamical systems controlling all body functions. Neurons are the building blocks of the nervous system and their impairment of their functions could result in neurodegenerative disorders. Accumulating evidence shows an increase of brain-affecting disorders, still today characterized by poor therapeutic options. There is a strong urgency to find new alternative strategies to prevent progressive neuronal loss. Polyphenols, a wide family of plant compounds with an equally wide range of biological activities, are suitable candidates to counteract chronic degenerative disease in the central nervous system. Herein, we will review their role in human healthcare and highlight their: antioxidant activities in reactive oxygen species-producing neurodegenerative pathologies; putative role as anti-acetylcholinesterase inhibitors; and protective activity in Alzheimer’s disease by preventing Aβ aggregation and tau hyperphosphorylation. Moreover, the pathology of these multifactorial diseases is also characterized by metal dyshomeostasis, specifically copper (Cu), zinc (Zn), and iron (Fe), most important for cellular function. In this scenario, polyphenols’ action as natural chelators is also discussed. Furthermore, the critical importance of the role exerted by polyphenols on microbiota is assumed, since there is a growing body of evidence for the role of the intestinal microbiota in the gut–brain axis, giving new opportunities to study molecular mechanisms and to find novel strategies in neurological diseases.

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“…The plates were washed with PBS and incubated in 1% FBS DMEM medium three consecutive times. The Ibidi culture system was also used in our experiments [20]. This was repeated three times for imaging measurements.…”

Section: Cell Migration Ability Detected By Wound Scratch Assaymentioning

confidence: 99%

Dihydroartemisinin inhibits EMT of glioma via gene BASP1 in extrachromosomal DNA

Que

Zhou

Liu

et al. 2023

Preprint

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The mechanism of dihydroartemisinin (DHA) inhibiting the migration and invasion of glioma in an ROS-DSB-dependent manner has been revealed. Extrachromosomal DNAs (ecDNAs) which are generated by DNA damage have great potential in glioma treatment. However, the role of ecDNAs in DHA pharmacological mechanisms in glioma is still unknown. In this study, DHA was found to inhibit proliferative activity, increase ROS levels and promote apoptosis in U87 and U251 cells. Migration and invasion have also been suppressed. ecDNA expression profiles were found in gliomas. EcDNA-BASP1 was found, by means of bioinformatics analysis, to be present in GBM tissues and positively correlated with patient prognosis. Proliferation, migration and invasion were upregulated after knockdown of ecDNA-BASP1. The expression of vimentin and N-cadherin also had the same tendency. Finally, we found that the ecDNA-BASP1 content in nude mouse transplant tumors was significantly increased after DHA treatment, which might exert a better suppressive effect on glioma. The upregulation of tumor suppressor ecDNA-BASP1 played an important role in the suppression of glioma progression induced by DHA. EcDNA-BASP1 may inhibit glioma migration and invasion through repressing epithelial-mesenchymal transition (EMT)

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Introducing Wound Healing Assays in the Undergraduate Biology Laboratory Using Ibidi Plates

Abstract: The wound healing assay is a simple and inexpensive method that allows researchers to experimentally mimic cell growth and migration leading to wound healing. In this assay, a wound is created on a monolayer of cultured mammalian cells and cell migration is monitored.

Cited by 5 publications

References 7 publications

Efficacy of metformin and electrical pulses in breast cancer MDA-MB-231 cells

Efficacy of metformin and electrical pulses in breast cancer MDA-MB-231 cells

Neuro-Nutraceutical Polyphenols: How Far Are We?

Dihydroartemisinin inhibits EMT of glioma via gene BASP1 in extrachromosomal DNA

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