Eleanor Stride

The cytoplasm of living cells behaves as a poroelastic material

The cytoplasm is the largest part of the cell by volume and hence its rheology sets the rate at which cellular shape changes can occur. Recent experimental evidence suggests that cytoplasmic rheology can be described by a poroelastic model, in which the cytoplasm is treated as a biphasic material consisting of a porous elastic solid meshwork (cytoskeleton, organelles, macromolecules) bathed in an interstitial fluid (cytosol). In this picture, the rate of cellular deformation is limited by the rate at which intracellular water can redistribute within the cytoplasm. However, direct supporting evidence for the model is lacking. Here we directly validate the poroelastic model to explain cellular rheology at physiologically relevant timescales using microindentation tests in conjunction with mechanical, chemical and genetic treatments. Our results show that water redistribution through the solid phase of the cytoplasm (cytoskeleton and macromolecular crowders) plays a fundamental role in setting cellular rheology.

Microbubble ultrasound contrast agents: A review:

Abstract The superior scattering properties of gas bubbles compared with blood cells have made microbubble ultrasound contrast agents important tools in ultrasound diagnosis. Over the past 2 years they have become the focus of a wide and rapidly expanding field of research, with their benefits being repeatedly demonstrated, both in ultrasound image enhancement, and more recently in drug and gene delivery applications. However, despite considerable investigation, their behaviour is by no means fully understood and, while no definite evidence of harmful effects has been obtained, there remain some concerns as to their safety. In this review the existing theoretical and experimental evidence is examined in order to clarify the extent to which contrast agents are currently understood and to identify areas for future research. In particular the disparity between the conditions considered in theoretical models and those encountered both in vitro, and more importantly in vivo is discussed, together with the controversy regarding the risk of harmful bio-effects.

Cavitation and contrast: The use of bubbles in ultrasound imaging and therapy

Abstract Microbubbles and cavitation are playing an increasingly significant role in both diagnostic and therapeutic applications of ultrasound. Microbubble ultrasound contrast agents have been in clinical use now for more than two decades, stimulating the development of a range of new contrast-specific imaging techniques which offer substantial benefits in echocardiography, microcirculatory imaging, and more recently, quantitative and molecular imaging. In drug delivery and gene therapy, microbubbles are being investigated/developed as vehicles which can be loaded with the required therapeutic agent, traced to the target site using diagnostic ultrasound, and then destroyed with ultrasound of higher intensity energy burst to release the material locally, thus avoiding side effects associated with systemic administration, e.g. of toxic chemotherapy. It has moreover been shown that the motion of the microbubbles increases the permeability of both individual cell membranes and the endothelium, thus enhancing therapeutic uptake, and can locally increase the activity of drugs by enhancing their transport across biologically inaccessible interfaces such as blood clots or solid tumours. In high-intensity focused ultrasound (HIFU) surgery and lithotripsy, controlled cavitation is being investigated as a means of increasing the speed and efficacy of the treatment. The aim of this paper is both to describe the key features of the physical behaviour of acoustically driven bubbles which underlie their effectiveness in biomedical applications and to review the current state of the art.

Novel microbubble preparation technologies

Gas microbubbles, stabilised by a surfactant or polymer coating, have become well established over the past 20–30 years as the most effective type of contrast agent available for ultrasound radiography. More recently, their potential for use in therapeutic applications including targeted drug delivery, gene therapy, thrombolysis and focused ultrasound surgery has also been investigated. Developments in both diagnostic and therapeutic applications have greatly increased the need for more advanced preparation technologies which provide a high degree of control over microbubble size, composition, stability and uniformity. Conventional processing techniques such as sonication and high shear emulsification offer high yield and low cost production but poor control over microbubble size and uniformity. This is being addressed by the development of new technologies, such as membrane emulsification, inkjet printing, electrohydrodynamic atomisation and microfluidic processing which offer significant improvements in terms of control over microbubble characteristics. The aim of this paper is to review the range of techniques available for microbubble preparation and how these have evolved to keep pace with advances in clinical practice.

Quantitative contrast-enhanced ultrasound imaging: a review of sources of variability

Ultrasound provides a valuable tool for medical diagnosis offering real-time imaging with excellent spatial resolution and low cost. The advent of microbubble contrast agents has provided the additional ability to obtain essential quantitative information relating to tissue vascularity, tissue perfusion and even endothelial wall function. This technique has shown great promise for diagnosis and monitoring in a wide range of clinical conditions such as cardiovascular diseases and cancer, with considerable potential benefits in terms of patient care. A key challenge of this technique, however, is the existence of significant variations in the imaging results, and the lack of understanding regarding their origin. The aim of this paper is to review the potential sources of variability in the quantification of tissue perfusion based on microbubble contrast-enhanced ultrasound images. These are divided into the following three categories: (i) factors relating to the scanner setting, which include transmission power, transmission focal depth, dynamic range, signal gain and transmission frequency, (ii) factors relating to the patient, which include body physical differences, physiological interaction of body with bubbles, propagation and attenuation through tissue, and tissue motion, and (iii) factors relating to the microbubbles, which include the type of bubbles and their stability, preparation and injection and dosage. It has been shown that the factors in all the three categories can significantly affect the imaging results and contribute to the variations observed. How these factors influence quantitative imaging is explained and possible methods for reducing such variations are discussed.

Generation of multilayered structures for biomedical applications using a novel tri-needle coaxial device and electrohydrodynamic flow

In this short communication, we describe the scope and flexibility of using a novel device containing three coaxially arranged needles to form a variety of novel morphologies. Different combinations of materials are subjected to controlled flow through the device under the influence of an applied electric field. The resulting electrohydrodynamic flow allows us to prepare double-layered bubbles, porous encapsulated threads and nanocapsules containing three layers. The ability to process such multilayered structures is very significant for biomedical engineering applications, for example, generating capsules for drug delivery, which can provide multistage controlled release.

A new method for the preparation of monoporous hollow microspheres.

The feasibility of producing a hollow microsphere with a single hole in its shell by coaxial electrohydrodynamic atomization (CEHDA) is demonstrated. Polymethylsilsesquioxane (PMSQ) was used as a model shell material encapsulating a core of a volatile liquid, perfluorohexane (PFH), which was subsequently evaporated to produce the hollow microspheres. The diameters of the microspheres and of the single surface pore were controlled by varying the flow rate of the components, the concentration of the PMSQ solution, and the applied voltage in the CEHDA process. The particles were characterized by scanning electron microscopy, and the ranges obtained were 275-860 nm for the microsphere diameter and 35-135 nm for the pore size. The process overcomes several of the key problems associated with existing methods of monoporous microsphere formation including removing the need for elevated temperatures, multiple processing steps, and the use of surfactants and other additives.

One-step electrohydrodynamic production of drug-loaded micro- and nanoparticles

The objective of this work was to produce drug-loaded nanometre- and micrometre-scale particles using a single-step process that provides control over particle size and size distribution. Co-axial electrohydrodynamic processing was used, at ambient temperature and pressure, with poly(lactic-co-glycolic acid) as the polymeric coating material and oestradiol as the encapsulated drug. The particle diameter was varied from less than 120 nm to a few micrometres, by simple methodical adjustments in the processing parameters (polymer concentration and applied voltage). In vitro studies were performed to determine the drug release profile from the particles during unassisted and ultrasound-stimulated degradation in simulated body fluid. An encapsulation efficiency of approximately 70% was achieved and release of the drug was sustained for a period of over 20 days. Exposing the particles to ultrasound (22.5 kHz) increased the rate of release by approximately 8 per cent. This processing method offers several advantages over conventional emulsification techniques for the preparation of drug-loaded particles. Most significantly, process efficiency and the drugs functionality are preserved, as complex multistep processing involving harsh solvents, other additives and elevated temperatures or pressures are avoided. Production rates of 1012 particles min−1 can be achieved with a single pair of co-axial needles and the process is amenable to being scaled up by using multiple sets.

Mapping microbubble viscosity using fluorescence lifetime imaging of molecular rotors

Encapsulated microbubbles are well established as highly effective contrast agents for ultrasound imaging. There remain, however, some significant challenges to fully realize the potential of microbubbles in advanced applications such as perfusion mapping, targeted drug delivery, and gene therapy. A key requirement is accurate characterization of the viscoelastic surface properties of the microbubbles, but methods for independent, nondestructive quantification and mapping of these properties are currently lacking. We present here a strategy for performing these measurements that uses a small fluorophore termed a “molecular rotor” embedded in the microbubble surface, whose fluorescence lifetime is directly related to the viscosity of its surroundings. We apply fluorescence lifetime imaging to show that shell viscosities vary widely across the population of the microbubbles and are influenced by the shell composition and the manufacturing process. We also demonstrate that heterogeneous viscosity distributions exist within individual microbubble shells even with a single surfactant component.

On the destruction of microbubble ultrasound contrast agents

In recent years, the use of microbubble ultrasound (US) contrast agents as carriers in drug and gene delivery applications has intensified the need for a clear understanding of the processes involved in their destruction. In this study, an analysis of the conditions in the shell of a contrast agent particle has been made, based on the full numerical solution of a modified Rayleigh-Plesset equation. The results indicate that extremely high shell stresses may be expected under typical clinical conditions. Examination of previous experimental evidence in the light of these findings suggests that the shells are almost invariably disrupted, even if they are not visibly destroyed. This has some serious implications, both for targeted delivery processes and reliable assessment of the potential for harmful bioeffects. At present, neither the model nor the experimental data provide an adequate description of contrast agent behaviour. This is due primarily to the lack of information regarding the mechanical response of the shell material and the restriction of the model to the case of small, spherically symmetrical oscillations. Methods for addressing these deficiencies in future work are proposed.