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In vitro measurement of attenuation and nonlinear scattering from echogenic liposomes
Authors:Paul Shirshendu  Russakow Daniel  Nahire Rahul  Nandy Tapas  Ambre Avinash H  Katti Kalpana  Mallik Sanku  Sarkar Kausik
Institution:a Mechanical Engineering, University of Delaware, Newark, DE 19716, United States
b North Dakota State University, Fargo, ND 58108, United States
c Mechanical and Aerospace Engineering, George Washington University, Washington, DC 20052, United States
Abstract:Echogenic liposomes (ELIP) are an excellent candidate for concurrent imaging and drug delivery applications. They combine the advantages of liposomes-biocompatibility and ability to encapsulate both hydrophobic and hydrophilic drugs-with strong reflections of ultrasound. The objective of this study is to perform a detailed in vitro acoustic characterization - including nonlinear scattering that has not been studied before - along with an investigation of the primary mechanism of echogenicity. Both components are critical for developing viable clinical applications of ELIP. Mannitol, a cryoprotectant, added during the preparation of ELIP is commonly believed to be critical in making them echogenic. Accordingly, here ELIP prepared with varying amount of mannitol concentration are investigated for their pressure dependent linear and non-linear scattered responses. The average diameter of these liposomes is measured to be 125-185 nm. But they have a broad size distribution including liposomes with diameters over a micro-meter as observed by TEM and AFM. These larger liposomes are critical for the overall echogenicity. Attenuation through liposomal solution is measured with four different transducers (central frequencies 2.25, 3.5, 5, 10 MHz). Measured attenuation increases linearly with liposome concentration indicating absence of acoustic interactions between liposomes. Due to the broad size distribution, the attenuation shows a flat response without a distinct peak in the range of frequencies (1-12 MHz) investigated. A 15-20 dB enhancement with 1.67 μg/ml of lipids is observed both for the scattered fundamental and the second harmonic responses at 3.5 MHz excitation frequency and 50-800 kPa amplitude. It demonstrates the efficacy of ELIP for fundamental as well as harmonic ultrasound imaging. The scattered response however does not show any distinct subharmonic peak for the acoustic excitation parameters studied. Small amount of mannitol proves critical for echogenicity. However, mannitol concentration above 100 mM shows no effect.
Keywords:Ultrasound imaging  Contrast agents  Echogenic liposomes  Scattering  Attenuation
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