In vivo performance of a microfabricated catheter for intraparenchymal delivery.

BACKGROUND: Convection-enhanced delivery (CED) is currently the only effective clinical technique to deliver biological therapeutic agents that would otherwise not cross the blood-brain barrier. Despite the promise of CED, several technical problems have limited its effectiveness. NEW METHOD: Brain infusions into a large mammal (pig) were performed with a catheter that was fabricated using micro-electro-mechanical systems (MEMS) technology (Olbricht et al., 2010). The performance of the catheter was evaluated for infusions at increasing infusion rates. Magnetic resonance (MR) images were acquired in real time to examine the distribution of infused tracers in the parenchyma. RESULTS: Both backflow and the distribution of CED of infusates into a variety of cytoarchitectures in porcine brain were quantified. Concentration profiles were determined for several MR contrast reagents as well as a fluorescent dye that are the sizes of small molecules, therapeutic proteins and an adeno-associated virus (AAV). The reagents can serve as surrogates for assessing the convective distribution of active molecules. Infusion rates up to 20µL/min were attained without evidence of backflow along the catheter. COMPARISON WITH EXISTING METHODS: The device performed well in terms of both backflow and infusion, superior to that of many studies reported in the literature on other catheters. All infused molecules had comparable ratios of distribution to infusion volumes. CONCLUSIONS: The catheter described in this report appears able to target tissue structures with precision, deliver therapeutics at high infusion rates, and resist backflow that can compromise the efficacy of CED therapy. The technology allows development of "smart" catheters for future applications.

Ultrasound-assisted convection-enhanced delivery to the brain in vivo with a novel transducer cannula assembly: laboratory investigation.

OBJECT: In convection-enhanced delivery (CED), drugs are infused locally into tissue through a cannula inserted into the brain parenchyma to enhance drug penetration over diffusion strategies. The purpose of this study was to demonstrate the feasibility of ultrasound-assisted CED (UCED) in the rodent brain in vivo using a novel, low-profile transducer cannula assembly (TCA) and portable, pocket-sized ultrasound system. METHODS: Forty Sprague-Dawley rats (350-450 g) were divided into 2 equal groups (Groups 1 and 2). Each group was divided again into 4 subgroups (n = 5 in each). The caudate of each rodent brain was infused with 0.25 wt% Evans blue dye (EBD) in phosphate-buffered saline at 2 different infusion rates of 0.25 µl/minute (Group 1), and 0.5 µl/minute (Group 2). The infusion rates were increased slowly over 10 minutes from 0.05 to 0.25 µl/minute (Group 1) and from 0.1 to 0.5 µl/minute (Group 2). The final flow rate was maintained for 20 minutes. Rodents in the 4 control subgroups were infused using the TCA without ultrasound and without and with microbubbles added to the infusate (CED and CED + MB, respectively). Rodents in the 4 UCED subgroups were infused without and with microbubbles added to the infusate (UCED and UCED + MB) using the TCA with continuous-wave 1.34-MHz low-intensity ultrasound at a total acoustic power of 0.11 ± 0.005 W and peak spatial intensity at the cannula tip of 49.7 mW/cm(2). An additional 4 Sprague-Dawley rats (350-450 g) received UCED at 4 different and higher ultrasound intensities at the cannula tip ranging from 62.0 to 155.0 mW/cm(2) for 30 minutes. The 3D infusion distribution was reconstructed using MATLAB analysis. Tissue damage and morphological changes to the brain were assessed using H & E. RESULTS: The application of ultrasound during infusion (UCED and UCED + MB) improved the volumetric distribution of EBD in the brain by a factor of 2.24 to 3.25 when there were no microbubbles in the infusate and by a factor of 1.16 to 1.70 when microbubbles were added to the infusate (p < 0.001). On gross and histological examination, no damage to the brain tissue was found for any acoustic exposure applied to the brain. CONCLUSIONS: The TCA and ultrasound device show promise to improve the distribution of infused compounds during CED. The results suggest further studies are required to optimize infusion and acoustic parameters for small compounds and for larger molecular weight compounds that are representative of promising antitumor agents. In addition, safe levels of ultrasound exposure in chronic experiments must be determined for practical clinical evaluation of UCED. Extension of these experiments to larger animal models is warranted to demonstrate efficacy of this technique.

In vivo two-photon excited fluorescence microscopy reveals cardiac- and respiration-dependent pulsatile blood flow in cortical blood vessels in mice.

Subtle alterations in cerebral blood flow can impact the health and function of brain cells and are linked to cognitive decline and dementia. To understand hemodynamics in the three-dimensional vascular network of the cerebral cortex, we applied two-photon excited fluorescence microscopy to measure the motion of red blood cells (RBCs) in individual microvessels throughout the vascular hierarchy in anesthetized mice. To resolve heartbeat- and respiration-dependent flow dynamics, we simultaneously recorded the electrocardiogram and respiratory waveform. We found that centerline RBC speed decreased with decreasing vessel diameter in arterioles, slowed further through the capillary bed, and then increased with increasing vessel diameter in venules. RBC flow was pulsatile in nearly all cortical vessels, including capillaries and venules. Heartbeat-induced speed modulation decreased through the vascular network, while the delay between heartbeat and the time of maximum speed increased. Capillary tube hematocrit was 0.21 and did not vary with centerline RBC speed or topological position. Spatial RBC flow profiles in surface vessels were blunted compared with a parabola and could be measured at vascular junctions. Finally, we observed a transient decrease in RBC speed in surface vessels before inspiration. In conclusion, we developed an approach to study detailed characteristics of RBC flow in the three-dimensional cortical vasculature, including quantification of fluctuations in centerline RBC speed due to cardiac and respiratory rhythms and flow profile measurements. These methods and the quantitative data on basal cerebral hemodynamics open the door to studies of the normal and diseased-state cerebral microcirculation.

Real-time imaging of perivascular transport of nanoparticles during convection-enhanced delivery in the rat cortex.

Convection-enhanced delivery (CED) is a promising technique for administering large therapeutics that do not readily cross the blood brain barrier to neural tissue. It is of vital importance to understand how large drug constructs move through neural tissue during CED to optimize construct and delivery parameters so that drugs are concentrated in the targeted tissue, with minimal leakage outside the targeted zone. Experiments have shown that liposomes, viral vectors, high molecular weight tracers, and nanoparticles infused into neural tissue localize in the perivascular spaces of blood vessels within the brain parenchyma. In this work, we used two-photon excited fluorescence microscopy to monitor the real-time distribution of nanoparticles infused in the cortex of live, anesthetized rats via CED. Fluorescent nanoparticles of 24 and 100 nm nominal diameters were infused into rat cortex through microfluidic probes. We found that perivascular spaces provide a high permeability path for rapid convective transport of large nanoparticles through tissue, and that the effects of perivascular spaces on transport are more significant for larger particles that undergo hindered transport through the extracellular matrix. This suggests that the vascular topology of the target tissue volume must be considered when delivering large therapeutic constructs via CED.

Cortical microhemorrhages cause local inflammation but do not trigger widespread dendrite degeneration.

Microhemorrhages are common in the aging brain, and their incidence is correlated with increased risk of neurodegenerative disease. Past work has shown that occlusion of individual cortical microvessels as well as large-scale hemorrhages can lead to degeneration of neurons and increased inflammation. Using two-photon excited fluorescence microscopy in anesthetized mice, we characterized the acute and chronic dynamics of vessel bleeding, tissue compression, blood flow change, neural degeneration, and inflammation following a microhemorrhage caused by rupturing a single penetrating arteriole with tightly-focused femtosecond laser pulses. We quantified the extravasation of red blood cells (RBCs) and blood plasma into the brain and determined that the bleeding was limited by clotting. The vascular bleeding formed a RBC-filled core that compressed the surrounding parenchymal tissue, but this compression was not sufficient to crush nearby brain capillaries, although blood flow speeds in these vessels was reduced by 20%. Imaging of cortical dendrites revealed no degeneration of the large-scale structure of the dendritic arbor up to 14 days after the microhemorrhage. Dendrites close to the RBC core were displaced by extravasating RBCs but began to relax back one day after the lesion. Finally, we observed a rapid inflammatory response characterized by morphology changes in microglia/macrophages up to 200 µm from the microhemorrhage as well as extension of cellular processes into the RBC core. This inflammation persisted over seven days. Taken together, our data suggest that a cortical microhemorrhage does not directly cause significant neural pathology but does trigger a sustained, local inflammatory response.

Fluid mechanics in the perivascular space.

Perivascular space (PVS) within the brain is an important pathway for interstitial fluid (ISF) and solute transport. Fluid flowing in the PVS can affect these transport processes and has significant impacts on physiology. In this paper, we carry out a theoretical analysis to investigate the fluid mechanics in the PVS. With certain assumptions and approximations, we are able to find an analytical solution to the problem. We discuss the physical meanings of the solution and particularly examine the consequences of the induced fluid flow in the context of convection-enhanced delivery (CED). We conclude that peristaltic motions of the blood vessel walls can facilitate fluid and solute transport in the PVS.

Fluid and solid mechanics in a poroelastic network induced by ultrasound.

We made a theoretical analysis on the fluid and solid mechanics in a poroelastic medium induced by low-power ultrasound. Using a perturbative approach, we were able to linearize the governing equations and obtain analytical solutions. We found that ultrasound could propagate in the medium as a mechanical wave, but would dissipate due to frictional forces between the fluid and the solid phase. The amplitude of the wave depends on the ultrasonic power input. We applied this model to the problem of drug delivery to soft biological tissues by low-power ultrasound and proposed a mechanism for enhanced drug penetration. We have also found the coexistence of two acoustic waves under certain circumstances and pointed out the importance of very accurate experimental determination of the high-frequency properties of brain tissue.

Microfluidic probes in the treatment of brain-related diseases.

Many new therapeutic compounds have been developed that target malignancies and other disorders of the brain. However, delivering these compounds to diseased tissue remains a difficult challenge. One option for local drug delivery in the brain is direct infusion of the compounds through a catheter into the brain parenchyma. Over the last decade, new infusion catheters have been developed to improve this delivery method. Some of these catheters are needles or cannulas that have been modified specifically to increase the infusion rate that can be achieved without leakage of the infusate out of the brain. Other new catheters have been fabricated using micromachining techniques adapted from electronics manufacturing. These microfabricated catheters can achieve comparable infusion rates as standard needles, but they also can incorporate features that would be difficult to build into needles or cannulas to improve drug delivery. This article reviews the development of these devices, their performance in preclinical studies and their potential benefits to neural drug delivery.

Retro-convection enhanced drug delivery: a computational study.

Retro-convection enhanced delivery (R-CED) is an emerging drug delivery method to overcome the blood brain barrier (BBB). We have developed a mathematical model to understand the fluid flow and mass transfer in the interstitium of brain tissue in R-CED therapy. The model was used to predict pressure distributions, fluid flow patterns, and drug concentration profiles. Some numerical results were obtained by computer simulations. Three kinds of microdialysis membranes used in R-CED protocols were analyzed in detail. While fluid flow was independent of the choice of membranes, mass transfer processes and drug distributions were found to be highly dependent on the choice of membranes. Sensitivity analysis on a variety of parameters and over a wide range of parameter values was carried out. Although R-CED turned out to be efficacious in generating fluid flows within the brain tissue, it did not favor a large effective treatment volume and needed to be re-examined and re-evaluated experimentally.

A portable high-intensity focused ultrasound device for noninvasive venous ablation.

BACKGROUND: Varicose veins and other vascular abnormalities are common clinical entities. Treatment options include vein stripping, sclerotherapy, and endovenous laser treatment, but all involve some degree of invasive intervention. The purpose of this study was to determine ex vivo the effectiveness of a novel hand-held, battery-operated, high-intensity focused ultrasound (HIFU) device for transcutaneous venous ablation. METHODS: The ultrasound device is 14 x 9 x 4 cm, weighs 650 g, and is powered by 4 lithium ion battery packs. An ex vivo testing platform consisting of two different models comprised of sequentially layered skin-muscle-vein or skin-fat-vein was developed, and specimens were treated with HIFU. The tissues were then disassembled, imaged, and processed for histology. The luminal cross-sectional area of vein that had been treated with HIFU and nontreated controls were measured, and the values presented as median and interquartile range (IQR). The values were compared using a Wilcoxon rank-sum test, and statistical significance was set at P < .05. RESULTS: On gross and histologic examination, veins that had been treated with HIFU showed evidence of coagulation necrosis. The surface of the muscle in direct contact with the vein had a pinpoint area of coagulation, whereas the adjacent fat appeared undisturbed; the skin, fat, and the surface of the muscle in contact with the transducer remained completely unaffected. The cross-sectional area was 3.79 mm(2) (IQR, 3.38-4.22) of the control vein lumen and 0.16 mm(2) (IQR, 0.04-0.39) in those that had been treated with HIFU (P = .0304). CONCLUSION: This inexpensive, portable HIFU device has the potential to allow clinicians to easily perform venous ablation in a manner that is entirely noninvasive and without the expense or inconvenience of large, complicated devices. This device represents a significant step forward in the development of new applications for HIFU technology.

Design and characterization of a high-power ultrasound driver with ultralow-output impedance.

We describe a pocket-sized ultrasound driver with an ultralow-output impedance amplifier circuit (less than 0.05 ohms) that can transfer more than 99% of the voltage from a power supply to the ultrasound transducer with minimal reflections. The device produces high-power acoustical energy waves while operating at lower voltages than conventional ultrasound driving systems because energy losses owing to mismatched impedance are minimized. The peak performance of the driver is measured experimentally with a PZT-4, 1.54 MHz, piezoelectric ceramic, and modeled using an adjusted Mason model over a range of transducer resonant frequencies. The ultrasound driver can deliver a 100 V(pp) (peak to peak) square-wave signal across 0-8 MHz ultrasound transducers in 5 ms bursts through continuous wave operation, producing acoustic powers exceeding 130 W. Effects of frequency, output impedance of the driver, and input impedance of the transducer on the maximum acoustic output power of piezoelectric transducers are examined. The small size, high power, and efficiency of the ultrasound driver make this technology useful for research, medical, and industrial ultrasonic applications.

Flexible microfluidic devices supported by biodegradable insertion scaffolds for convection-enhanced neural drug delivery.

Convection enhanced delivery (CED) can improve the spatial distribution of drugs delivered directly to the brain. In CED, drugs are infused locally into tissue through a needle or catheter inserted into brain parenchyma. Transport of the infused material is dominated by convection, which enhances drug penetration into tissue compared with diffusion mediated delivery. We have fabricated and characterized an implantable microfluidic device for chronic convection enhanced delivery protocols. The device consists of a flexible parylene-C microfluidic channel that is supported during its insertion into tissue by a biodegradable poly(DL-lactide-co-glycolide) scaffold. The scaffold is designed to enable tissue penetration and then erode over time, leaving only the flexible channel implanted in the tissue. The device was able to reproducibly inject fluid into neural tissue in acute experiments with final infusate distributions that closely approximate delivery from an ideal point source. This system shows promise as a tool for chronic CED protocols.

Development of a portable therapeutic and high intensity ultrasound system for military, medical, and research use.

We have developed a portable high power ultrasound system with a very low output impedance amplifier circuit (less than 0.3 Omega) that can transfer more than 90% of the energy from a battery supply to the ultrasound transducer. The system can deliver therapeutic acoustical energy waves at lower voltages than those in conventional ultrasound systems because energy losses owing to a mismatched impedance are eliminated. The system can produce acoustic power outputs over the therapeutic range (greater then 50 W) from a PZT-4, 1.54 MHz, and 0.75 in diameter piezoelectric ceramic. It is lightweight, portable, and powered by a rechargeable battery. The portable therapeutic ultrasound unit has the potential to replace "plug-in" medical systems and rf amplifiers used in research. The system is capable of field service on its internal battery, making it especially useful for military, ambulatory, and remote medical applications.

Dilation and degradation of the brain extracellular matrix enhances penetration of infused polymer nanoparticles.

This study investigates methods of manipulating the brain extracellular matrix (ECM) to enhance the penetration of nanoparticle drug carriers in convection-enhanced delivery (CED). A probe was fabricated with two independent microfluidic channels to infuse, either simultaneously or sequentially, nanoparticles and ECM-modifying agents. Infusions were performed in the striatum of the normal rat brain. Monodisperse polystyrene particles with a diameter of 54 nm were used as a model nanoparticle system. Because the size of these particles is comparable to the effective pore size of the ECM, their transport may be significantly hindered compared with the transport of low molecular weight molecules. To enhance the transport of the infused nanoparticles, we attempted to increase the effective pore size of the ECM by two methods: dilating the extracellular space and degrading selected constituents of the ECM. Two methods of dilating the extracellular space were investigated: co-infusion of nanoparticles and a hyperosmolar solution of mannitol, and pre-infusion of an isotonic buffer solution followed by infusion of nanoparticles. These treatments resulted in an increase in the nanoparticle distribution volume of 51% and 123%, respectively. To degrade hyaluronan, a primary structural component of the brain ECM, a pre-infusion of hyaluronidase (20,000 U/mL) was followed after 30 min by infusion of nanoparticles. This treatment resulted in an increase in the nanoparticle distribution of 64%. Our results suggest that both dilation and enzymatic digestion can be incorporated into CED protocols to enhance nanoparticle penetration.

Rapid self-assembly of core-shell organosilicon microcapsules within a microfluidic device.

The preparation of hierarchically structured organosilicon microcapsules from commercially available starting materials is described. Using a microfluidic device, an emulsion of dichlorodiphenylsilane is formed in a continuous phase of aqueous glycerol. The silane droplets undergo hydrolysis, condensation, and crystallization within minutes to form self-assembled, core-shell microcapsules. The microparticles have been characterized with light and electron microscopy, nuclear magnetic resonance spectroscopy (NMR), diffusion-ordered NMR spectroscopy (DOSY), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and powder X-ray diffraction (XRD). The characterization data show that the microcapsule walls consist of amorphous, oligomeric poly(diphenylsiloxane) surrounded by a spiny layer of crystalline diphenylsilanediol. Glycerol is occluded within the wall material but is not covalently bound to the silicon components. Glycerol is a crucial element for producing low-dispersity microcapsules with well-ordered surface spines, as the use of methyl cellulose as viscomodifier yields amorphous surfaces.

Building drug delivery into tissue engineering.

The creation of efficient methods for manufacturing biotechnology drugs--many of which influence fundamental but complex cell behaviours, such as proliferation, migration and differentiation--is creating new opportunities for tissue repair. Many agents are potent and multifunctional; that is, they produce different effects within different tissues. Therefore, control of tissue concentration and spatial localization of delivery is essential for safety and effectiveness. Synthetic systems that can control agent delivery are particularly promising as materials for enhancing tissue regeneration. This review discusses the state of the art in controlled-release and microfluidic drug delivery technologies, and outlines their potential applications for tissue engineering.