title: Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro.
abstract: This work is focused on the in vitro study of the effects induced by medical ultrasound (US) in murine fibroblast cells (NIH-3T3) at a low-intensity of exposure (spatial peak temporal average intensity Ita<0.1Wcm(-2)). Conventional 1MHz and 3MHz US devices of therapeutic relevance were employed with varying intensity and exposure time parameters. In this framework, upon cells exposure to US, structural changes at the molecular level were evaluated by infrared spectroscopy; alterations in plasma membrane permeability were monitored in terms of uptake efficiency of small cell-impermeable model drug molecules, as measured by fluorescence microscopy and flow cytometry. The results were related to the cell viability and combined with the statistical PCA analysis, confirming that NIH-3T3 cells are sensitive to therapeutic US, mainly at 1MHz, with time-dependent increases in both efficiency of uptake, recovery of wild-type membrane permeability, and the size of molecules entering 3T3. On the contrary, the exposures from US equipment at 3MHz show uptakes comparable with untreated samples.
authors.affiliation: Sapienza University, Physics Department, Rome, Italy. Electronic address: fabiodomenici@gmail.com.
authors.forename: Fabio
authors.initials: F
authors.lastname: Domenici
authors.affiliation: INAIL, Italian Workers' Compensation Authority, Rome, Italy. Electronic address: c.giliberti@inail.it.
authors.forename: Claudia
authors.initials: C
authors.lastname: Giliberti
authors.affiliation: INAIL, Italian Workers' Compensation Authority, Rome, Italy.
authors.forename: Angelico
authors.initials: A
authors.lastname: Bedini
authors.affiliation: INAIL, Italian Workers' Compensation Authority, Rome, Italy.
authors.forename: Raffaele
authors.initials: R
authors.lastname: Palomba
authors.affiliation: Roma Tre University, Department of Science, Rome, Italy.
authors.forename: Ion
authors.initials: I
authors.lastname: Udroiu
authors.affiliation: Sapienza University, Physics Department, Rome, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Di Giambattista
authors.affiliation: Sapienza University, Department of Molecular Medicine, Rome, Italy.
authors.forename: Deleana
authors.initials: D
authors.lastname: Pozzi
authors.affiliation: Sapienza University, Department of Experimental Medicine, Rome, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Morrone
authors.affiliation: Sapienza University, Physics Department, Rome, Italy.
authors.forename: Federico
authors.initials: F
authors.lastname: Bordi
authors.affiliation: Sapienza University, Physics Department, Rome, Italy. Electronic address: agostina.congiu@gmail.com.
authors.forename: Agostina
authors.initials: A
authors.lastname: Congiu Castellano
mesh_terms: D000818:Animals; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology*; D005434:Flow Cytometry; D005452:Fluoresceins / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D008856:Microscopy, Fluorescence; D013055:Spectrophotometry, Infrared; D014465:Ultrasonics / Q000379:methods*
chemical_list: D005452:Fluoresceins; C007740:fluorexon
keywords: Cell uptake; FTIR spectroscopy; Fluorescence microscopy; Membrane sonoporation; Ultrasound