The Synergistic Effects of Ultrasound Waves and Ethanol Exposure on Intracellular Calcium and Reactive Oxygen Species Rates of Hippocampal Cells During the Embryonic Course in Rats

Ranaei, Elahe; Abbasi, Naser; Moayeri, Ardeshir; Kafashi, Mina; Maleki, Maryam; Kaffashian, Mohammad Reza

doi:10.4274/tnd.2022.33270

Cited by 1 publication

(1 citation statement)

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“…This is in contrast with the observations by others, although this discrepancy could be explained due to different US properties. Indeed, in our experiments, we used 12 MHz frequencies instead of the commonly used 1-5 MHz [48,50,51]. Moreover, US treatment has been reported to trigger the ER-stress response, leading to the production of GRP78 [49].…”

Section: Discussionmentioning

confidence: 99%

Morphological and Functional Effects of Ultrasound on Blood–Brain Barrier Transitory Opening: An In Vitro Study on Rat Brain Endothelial Cells

Branca

Boninsegna

Morucci

et al. 2023

Cells

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With the recent advances in medicine, human life expectancy is increasing; however, the extra years of life are not necessarily spent in good health or free from disability, resulting in a significantly higher incidence of age-associated pathologies. Among these disorders, neurodegenerative diseases have a significant impact. To this end, the presence of the protective blood–brain barrier (BBB) represents a formidable obstacle to the delivery of therapeutics. Thus, this makes it imperative to define strategies to bypass the BBB in order to successfully target the brain with the appropriate drugs. It has been demonstrated that targeting the BBB by ultrasound (US) can transiently make this anatomical barrier permeable and in so doing, allow the delivery of therapeutics. Thus, our aim was to carry out an in depth in vitro molecular and morphological study on the effects of US treatment on the BBB. The rat brain endothelial (RBE4) cell line was challenged with exposure to 12 MHz diagnostic US treatment for 10, 20, and 30 min. Cell viability assays, Western blotting analysis on the endoplasmic reticulum (ER), and oxidative stress marker evaluation were then performed, along with cytological and immunofluorescence staining, in order to evaluate the effects of US on the intercellular spaces and tight junction distribution of the brain endothelial cells. We observed that the US treatment exerted no toxic effects on either RBE4 cell viability or the upregulation/dislocation of the ER and oxidative stress marker (GRP78 and cytochrome C, respectively). Further, we observed that the application of US induced an increase in the intercellular spaces, as shown by Papanicolaou staining, mainly due to the altered distribution of the tight junction protein zonula occludens-1 (ZO-1). This latter US-dependent effect was transient and disappeared 20 min after the removal of the stimulus. In conclusion, our results show that US induces a transient alteration of the BBB, without altering the intracellular signaling pathways such as the ER and oxidative stress that could potentially be toxic for endothelial cells. These results suggested that US treatment could represent a potential strategy for improving drug delivery to the brain.

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