Phospholipid-Coated Gas Bubble Engineering: Key Parameters for Size and Stability Control, as Determined by an Acoustical Method

Abstract

We have recently reported the sampling of differently sized monomodal populations of microbubbles from a polydisperse lipid-coated bubble preparation. The microbubbles were coated with dimyristoylphosphatidylcholine (DMPC) and stabilized by perfluorohexane (PFH). Such microbubbles are useful as contrast agents and, potentially, for oxygen, drug, and gene delivery and as therapeutic devices. Monomodal populations of small bubbles (approximately 1.6 microm in radius) and large bubbles (approximately 5.4 microm) have been obtained, as assessed by acoustical measurement, static light scattering, and optical microscopy. In this paper, we have determined the influence of various preparation parameters on the initial size characteristics (mean radius and radii distribution) of the microbubbles and on their stability upon time. The bubble size was determined acoustically, with a homemade acoustic setup equipped with a low-power emitter, to avoid altering the bubble stability. We have focused on the effects of the bubble flotation time during the fractionation process and on the DMPC concentration. PFH was indispensable for obtaining stable bubbles. The nature of the buffer [Isoton II vs N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)] used as the continuous phase did not significantly impact the bubble characteristics and stability. In both buffers, the half-lives of small bubbles (approximately 1.6 microm in radius in Isoton II and approximately 2.1 microm in HEPES) were found to be longer than those of larger ones (approximately 5.4 and approximately 5.9 microm in Isoton II and HEPES, respectively). The bubble stability study revealed that in both buffers, the average radius of the population of large bubbles progressively increased with time. On the other hand, the average radius of the population of small bubbles decreased slightly in Isoton II and remained constant in HEPES. This suggests that the dissolution behavior of small and large bubbles is governed by different mechanisms.