Pulmonary tuberculosis (TB), which is caused by Mycobacterium tuberculosis (Mtb), remains a global pandemic, despite the widespread use of the parenteral live attenuated Bacillus Calmette–Guérin (BCG) vaccine during the past decades. Mucosal administration of next generation TB vaccines has great potential, but developing a safe and efficacious mucosal vaccine is challenging. Hence, understanding the in vivo biodistribution and pharmacokinetics of mucosal vaccines is essential for shaping the desired immune response and for optimal spatiotemporal targeting of the appropriate effector cells in the lungs. A subunit vaccine consisting of the fusion antigen H56 (Ag85B-ESAT-6-Rv2660) and the liposome-based cationic adjuvant formulation (CAF01) confers efficient protection in preclinical animal models. In this study, we devise a novel immunization strategy for the H56/CAF01 vaccine, which comply with the intrapulmonary (i.pulmon.) route of immunization. We also describe a novel dual-isotope (111In/67Ga) radiolabeling approach, which enables simultaneous non-invasive and longitudinal SPECT/CT imaging and quantification of H56 and CAF01 upon parenteral prime and/or i.pulmon. boost immunization. Our results demonstrate that the vaccine is distributed evenly in the lungs, and there are pronounced differences in the pharmacokinetics of H56 and CAF01. We provide convincing evidence that the H56/CAF01 vaccine is not only well-tolerated when administered to the respiratory tract, but it also induces strong lung mucosal and systemic IgA and polyfunctional Th1 and Th17 responses after parenteral prime and i.pulmon. boost immunization. The study furthermore evaluate the application of SPECT/CT imaging for the investigation of vaccine biodistribution after parenteral and i.pulmon. immunization of mice.

No abstract

Droplet microfluidics technology has recently been introduced to generate particles for many biomedical applications that include therapeutic embolizing agents in hepatic, uterine or bronchial arteries. Embolic agents are available in a variety of shapes and sizes that are adjusted according to the target vessel characteristics. Magnetic embolic agents can additionally be navigated to the target location (e.g., a tumor) through the blood system by applying an external magnetic field. This technology is termed Magnetic Resonance Navigation (MRN). Here we introduce a high throughput method to produce homogeneously sized magnetic microspheres (MMS) as blood vessel embolic agents for use in combination with MRN. The system for MMS production consists of a simple 3D printed micro coflowing device that is able to produce biocompatible, degradation rate controllable poly­(lactic-co-glycolic acid) (PLGA) microspheres encasing magnetic nanoparticles. Axisymmetric flow is obtained with a central needle injecting the dispersed phase surrounded by a continuous phase and leads to the formation of size-controlled droplets that turn into homogeneously sized MMS linearly dependent on the inner needle diameter. MMS morphology, mean particle size and size distribution were quantified from SEM images. Magnetic performance of MMS was investigated using a vibrating sample magnetometer. MMS were nontoxic toward HUVEC (human umbilical vein endothelial cells) and HEK293 (human embryonic kidney) cells. The presented micro coflowing method allows for the reliable production of large MMS sized 130–700 μm with narrow size distribution (CV < 7%) and magnetic properties useful for MRN.

Purpose The purpose of this study was to demonstrate the feasibility of using a custom gradient sequence on an unmodified 3T magnetic resonance imaging (MRI) scanner to perform magnetic resonance navigation (MRN) by investigating the blood flow control method in vivo, reproducing the obtained rheology in a phantom mimicking porcine hepatic arterial anatomy, injecting magnetized microbead aggregates through an implantable catheter, and steering the aggregates across arterial bifurcations for selective tumor embolization. Materials and Methods In the first phase, arterial hepatic velocity was measured using cine phase‐contrast imaging in seven pigs under free‐flow conditions and controlled‐flow conditions, whereby a balloon catheter is used to occlude arterial flow and saline is injected at different rates. Three of the seven pigs previously underwent selective lobe embolization to simulate a chemoembolization procedure. In the second phase, the measured in vivo controlled‐flow velocities were approximately reproduced in a Y‐shaped vascular bifurcation phantom by injecting saline at an average rate of 0.6 mL/s with a pulsatile component. Aggregates of 200‐μm magnetized particles were steered toward the right or left hepatic branch using a 20‐mT/m MRN gradient. The phantom was oriented at 0°, 45°, and 90° with respect to the B0 magnetic field. The steering differences between left–right gradient and baseline were calculated using Fisher's exact test. A theoretical model of the trajectory of the aggregate within the main phantom branch taking into account gravity, magnetic force, and hydrodynamic drag was also designed, solved, and validated against the experimental results to characterize the physical limitations of the method. Results At an injection rate of 0.5 mL/s, the average flow velocity decreased from 20 ± 15 to 8.4 ± 5.0 cm/s after occlusion in nonembolized pigs and from 13.6 ± 2.0 to 5.4 ± 3.0 cm/s in previously embolized pigs. The pulsatility index measured to be 1.7 ± 1.8 and 1.1 ± 0.1 for nonembolized and embolized pigs, respectively, decreased to 0.6 ± 0.4 and 0.7 ± 0.3 after occlusion. For MRN performed at each orientation, the left–right distribution of aggregates was 55%, 25%, and 75% on baseline and 100%, 100%, and 100% (P < 0.001, P = 0.003, P = 0.003) after the application of MRN, respectively. According to the theoretical model, the aggregate reaches a stable transverse position located toward the direction of the gradient at a distance equal to 5.8% of the radius away from the centerline within 0.11 s, at which point the aggregate will have transited through a longitudinal distance of 1.0 mm from its release position. Conclusion In this study, we showed that the use of a balloon catheter reduces arterial hepatic flow magnitude and variation with the aim to reduce steering failures caused by fast blood flow rates and low magnetic steering forces. A mathematical model confirmed that the reduced flow rate is low enough to maximize steering ratio. After reproducing the flow rate in a vascular bifur...

No abstract

Background The discovery and development of new medicines requires high-throughput screening of possible therapeutics in a specific model of the disease. Infrared thermal imaging (IRT) is a modern assessment method with extensive clinical and preclinical applications. Employing IRT in longitudinal preclinical setting to monitor arthritis onset, disease activity and therapeutic efficacies requires a standardized framework to provide reproducible quantitative data as a precondition for clinical studies. Methods Here, we established the accuracy and reliability of an inexpensive smartphone connected infrared (IR) camera against known temperature objects as well as certified blackbody calibration equipment. An easy to use protocol incorporating contactless image acquisition and computer-assisted data analysis was developed to detect disease-related temperature changes in a collagen-induced arthritis (CIA) mouse model and validated by comparison with two conventional methods, clinical arthritis scoring and paw thickness measurement. We implemented IRT to demonstrate the beneficial therapeutic effect of nanoparticle drug delivery versus free methotrexate (MTX) in vivo. Results The calibrations revealed high accuracy and reliability of the IR camera for detecting temperature changes in the rheumatoid arthritis animal model. Significant positive correlation was found between temperature changes and paw thickness measurements as the disease progressed. IRT was found to be superior over the conventional techniques specially at early arthritis onset, when it is difficult to observe subclinical signs and measure structural changes. Conclusion IRT proved to be a valid and unbiased method to detect temperature changes and quantify the degree of inflammation in a rapid and reproducible manner in longitudinal preclinical drug efficacy studies.

No abstract

Objec0ve: Superparamagne0c nanopar0cles (SPIONs) can be combined with tumor chemoemboliza-0on agents to form magne0c drug-elu0ng beads (MDEBs), which are navigated magne0cally in the MRI scanner through the vascular system. We aim to develop a method to accurately quan0fy and localize these par0cles and to validate the method in phantoms and swine models. Meth-ods: MDEBs were made of Fe3O4 SPIONs. Aaer injected known numbers of MDEBs, suscep0bility ar0facts in three-dimensional (3D) volumetric interpolated breath-hold ex-amina0on (VIBE) sequences were acquired in glass and Polyvinyl alcohol (PVA) phantoms, and two living swine. Image processing of VIBE images provided the volume rela0onship between MDEBs and their ar0fact at different VIBE acquisi0ons and post-processing parameters. Sim-ulated hepa0c-artery emboliza0on was performed in vivo with an MRI-condi0onal magne0c-injec0on system, using the volume rela0onship to locate and quan0fy MDEB distri-bu0on. Results: Individual MDEBs were spa0ally identified, and their ar0facts quan0fied, showing no correla0on with magne0c-field orienta0on or sequence bandwidth, but ex-hibi0ng a rela0onship with echo 0me and providing a lin-ear volume rela0onship. Two MDEB aggregates were mag-ne0cally steered into desired liver regions while the other 19 had no steering, and 25 aggregates were injected into another swine without steering. The MDEBs were spa0ally iden0fied and the volume rela0onship showed accuracy in assessing the number of the MDEBs, with small errors (≤ 8.8%). Conclusion and Significance: MDEBs were able to be steered into desired body regions and then localized using 3D VIBE sequences. The resul0ng volume rela0onship was linear, robust, and allowed for quan0ta0ve analysis of the MDEB distribu0on.