Preparation and Characterization of PLA Ultrasound Contrast Agents by Combining an Ultrasound Method and a Shirasu Porous Glass (SPG) Membrane Emulsification Technique

Abstract

The ultrasound contrast agent based on a poly lactic acid (PLA) was prepared by combining an ultrasound method and a Shirasu Porous Glass (SPG) membrane emulsification technique. An aqueous phase containing ammonium bicarbonate was used as the internal water phase (Wi), and PLA and Span 80 were dissolved in a solvent of dichloromethane (DCM), which was used as the oil phase (O). These two solutions were probe sonicated to form a Wi/O primary emulsion. The primary emulsion was permeated through the uniform pores (1.1 mum) of an SPG membrane into the external water phase (W <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> ) by the pressure of nitrogen gas to form the uniform W <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">1</sub> /O/W <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">2</sub> droplets. After DCM was evaporated, the hardened PLA microcapsules were further formulated into a lyophilized powder containing decafluorobutane gas. SEM image demonstrated that the PLA microcapsules were sphere-shaped and internally hollow with an average diameter ranging from 1.99 to 3.58 mum. In vitro, the PLA contrast agents showed high acoustic enhancement properties, the enhancement increased nonlinearly with dose, and the minimal loss (less than 5 dB) of signal was observed over 20 min of analysis at 37degC, the maximum acoustic enhancement was 45 dB, which significantly higher (p < 0.01) compared to a value of 28 dB for those prepared by a conventional solvent evaporation method. In conclusion, the hollow PLA microcapsules prepared by the novel method have the characteristics desirable for an intravenously administered ultrasound contrast agents.