Switching between magnetotactic and aerotactic displacement controls to enhance the efficacy of MC-1 magneto-aerotactic bacteria as cancer-fighting nanorobots

The delivery of drug molecules to tumor hypoxic areas could yield optimal therapeutic outcomes. This suggests that effective cancer-fighting micro- or nanorobots would require more integrated functionalities than just the development of directional propelling constructs which have so far been the main general emphasis in medical micro- and nanorobotic research. Development of artificial agents that would be most effective in targeting hypoxic regions may prove to be a very challenging task considering present technological constraints. Self-propelled, sensory-based and directionally-controlled agents in the form of Magnetotactic Bacteria (MTB) of the MC-1 strain have been investigated as effective therapeutic nanorobots in cancer therapy. Following computer-based magnetotactic guidance to reach the tumor area, the microaerophilic response of drug-loaded MC-1 cells could be exploited in the tumoral interstitial fluid microenvironments. Accordingly, their swimming paths would be guided by a decreasing oxygen concentration towards the hypoxic regions. However, the implementation of such a targeting strategy calls for a method to switch from a computer-assisted magnetotactic displacement control to an autonomous aerotactic displacement control. In this way, the MC-1 cells will navigate to tumoral regions and, once there, target hypoxic areas through their microaerophilic behavior. Here we show not only how the magnitude of the magnetic field can be used for this purpose but how the findings could help determine the specifications of a future compatible interventional platform within known technological and medical constraints

Using the Fringe Field of MRI Scanner for the Navigation of Microguidewires in the Vascular System

Le traitement du cancer, la prévention des accidents vasculaires cérébraux et le diagnostic ou le traitement des maladies vasculaires périphériques sont tous des cas d'application d'interventions à base de cathéter par le biais d'un traitement invasif minimal. Cependant, la pratique du cathétérisme est généralement pratiquée manuellement et dépend fortement de l'expérience et des compétences de l'interventionniste. La robotisation du cathétérisme a été étudiée pour faciliter la procédure en augmentant les niveaux d’autonomie par rapport à cette pratique clinique. En ce qui concerne ce problème, un des problèmes concerne le placement super sélectif du cathéter dans les artères plus étroites nécessitant une miniaturisation de l'instrument cathéter / fil de guidage attaché. Un microguide qui fonctionne dans des vaisseaux sanguins étroits et tortueux subit différentes forces mécaniques telles que le frottement avec la paroi du vaisseau. Ces forces peuvent empêcher la progression de la pointe du fil de guidage dans les vaisseaux. Une méthode proposée consiste à appliquer une force de traction à la pointe du microguide pour diriger et insérer le dispositif tout en poussant l’instrument attaché à partir de l’autre extrémité n’est plus pratique, et à exploiter le gradient du champ de franges IRM surnommé Fringe Field Navigation (FFN ) est proposée comme solution pour assurer cet actionnement. Le concept de FFN repose sur le positionnement d'un patient sur six DOF dans le champ périphérique du scanner IRM afin de permettre un actionnement directionnel pour la navigation du fil-guide. Ce travail rend compte des développements requis pour la mise en oeuvre de la FFN et l’étude du potentiel et des possibilités qu’elle offre au cathétérisme, en veillant au renforcement de l’autonomie. La cartographie du champ de franges d'un scanner IRM 3T est effectuée et la structure du champ de franges en ce qui concerne son uniformité locale est examinée. Une méthode pour la navigation d'un fil de guidage le long d'un chemin vasculaire souhaité basée sur le positionnement robotique du patient à six DOF est développée. Des expériences de FFN guidées par rayons X in vitro et in vivo sur un modèle porcin sont effectuées pour naviguer dans un fil de guidage dans la multibifurcation et les vaisseaux étroits. Une caractéristique unique de FFN est le haut gradient du champ magnétique. Il est démontré in vitro et in vivo que cette force surmonte le problème de l'insertion d'un fil microguide dans des vaisseaux tortueux et étroits pour permettre de faire avancer le fil-guide avec une distale douce au-delà de la limite d'insertion manuelle. La robustesse de FFN contre les erreurs de positionnement du patient est étudiée en relation avec l'uniformité locale dans le champ périphérique. La force élevée du champ magnétique disponible dans le champ de franges IRM peut amener les matériaux magnétiques doux à son état de saturation. Ici, le concept d'utilisation d'un ressort est présenté comme une alternative vi déformable aux aimants permanents solides pour la pointe du fil-guide. La navigation d'un microguide avec une pointe de ressort en structure vasculaire complexe est également réalisée in vitro. L'autonomie de FFN en ce qui concerne la planification d'une procédure avec autonomie de tâche obtenue dans ce travail augmente le potentiel de FFN en automatisant certaines étapes d'une procédure. En conclusion, FFN pour naviguer dans les microguides dans la structure vasculaire complexe avec autonomie pour effectuer le positionnement du patient et contrôler l'insertion du fil de guidage - avec démonstration in vivo dans un modèle porcin - peut être considéré comme un nouvel outil robotique facilitant le cathétérisme vasculaire. tout en aidant à cibler les vaisseaux lointains dans le système vasculaire.----------ABSTRACT Treatment of cancer, prevention of stroke, and diagnosis or treatment of peripheral vascular diseases are all the cases of application of catheter-based interventions through a minimal-invasive treatment. However, performing catheterization is generally practiced manually, and it highly depends on the experience and the skills of the interventionist. Robotization of catheterization has been investigated to facilitate the procedure by increasing the levels of autonomy to this clinical practice. Regarding it, one issue is the super selective placement of the catheter in the narrower arteries that require miniaturization of the tethered catheter/guidewire instrument. A microguidewire that operates in narrow and tortuous blood vessels experiences different mechanical forces like friction with the vessel wall. These forces can prevent the advancement of the tip of the guidewire in the vessels. A proposed method is applying a pulling force at the tip of the microguidewire to steer and insert the device while pushing the tethered instrument from the other end is no longer practical, and exploiting the gradient of the MRI fringe field dubbed as Fringe Field Navigation (FFN) is proposed as a solution to provide this actuation. The concept of FFN is based on six DOF positioning of a patient in the fringe field of the MRI scanner to enable directional actuation for the navigation of the guidewire. This work reports on the required developments for implementing FFN and investigating the potential and the possibilities that FFN introduces to the catheterization, with attention to enhancing the autonomy. Mapping the fringe field of a 3T MRI scanner is performed, and the structure of the fringe field regarding its local uniformity is investigated. A method for the navigation of a guidewire along a desired vascular path based on six DOF robotic patient positioning is developed. In vitro and in vivo x-ray Guided FFN experiments on a swine model of are performed to navigate a guidewire in the multibifurcation and narrow vessels. A unique feature of FFN is the high gradient of the magnetic field. It is demonstrated in vitro and in vivo that this force overcomes the issue of insertion of a microguidewire in tortuous and narrow vessels to enable advancing the guidewire with a soft distal beyond the limit of manual insertion. Robustness of FFN against the error in the positioning of the patient is investigated in relation to the local uniformity in the fringe field. The high strength of the magnetic field available in MRI fringe field can bring soft magnetic materials to its saturation state. Here, the concept of using a spring is introduced as a deformable alternative to solid permanent magnets for the tip of the guidewire. Navigation of a microguidewire with a viii spring tip in complex vascular structure is also performed in vitro. The autonomy of FFN regarding planning a procedure with Task Autonomy achieved in this work enhances the potential of FFN by automatization of certain steps of a procedure. As a conclusion, FFN to navigate microguidewires in the complex vascular structure with autonomy in performing tasks of patient positioning and controlling the insertion of the guidewire – with in vivo demonstration in swine model – can be considered as a novel robotic tool for facilitating the vascular catheterization while helping to target remote vessels in the vascular system

Building a Magnetosome-like Nanoparticle for Magnetic Resonance Imaging

With its superb spatial and temporal resolution, magnetic resonance imaging (MRI) has great potential to track cellular activities that define early stages of disease. To improve molecular imaging techniques, we are developing MRI reporter gene expression based on the magnetosome. In magnetotactic bacteria (MTB), magnetosome formation compartmentalizes iron biominerals in membrane-enclosed vesicles. We hypothesize that essential magnetosome proteins interact in any cell type to form rudimentary magnetosome-like nanoparticles, providing a genetically-controlled contrast agent for molecular MRI. MTB genes mamE, mamB, mamI, and mamL were cloned from M. magneticum sp. AMB-1 genomic DNA by PCR and inserted into fluorescent vectors to create Mam fusion proteins then stably expressed in human MDA-MB-435 melanoma cells. Cines of fluorescent elements detected in intact cells were captured with confocal microscopy (Nikon A1R) and analyzed using both ImageJ and Mathematica for Brownian motion and velocity. To obtain longitudinal and transverse relaxation rates, cells stably expressing magnetosome proteins were supplemented with 250 µM ferric nitrate, harvested, mounted in a gelatin phantom, and scanned at 3 Tesla (Biograph mMR). Tomato-MamL, Tomato-MamL/GFP-MamI, and Tomato-MamB all express punctate, mobile fluorescence, while GFP-MamE expresses punctate but stationary fluorescence. Analysis of motility revealed that magnetosome proteins have variable diffusion coefficients due to their variable sizes, but all magnetosome proteins travel at a velocity of around 0.2 µm/s. Relaxation rates of iron-supplemented cells expressing Tomato-MamB, GFP-MamI, or Tomato-MamL have significantly higher R2 and R2* than non-supplemented cells. Interestingly, iron-supplemented cells expressing GFP-MamE or co-expressing FLAG-MamL/GFP-MamI had relaxation rates comparable to unsupplemented cell types. This is the first report characterizing essential magnetosome proteins MamE, MamB, MamI, and MamL in mammalian cells. Analysis of motion shows that magnetosome proteins travel at velocities comparable to the mammalian motor protein myosin. Expression of either MamB, MamI or MamL increases transverse relaxation rates; however, co-expression of MamI and MamL reduces them again, suggesting a regulatory effect of magnetosome gene combinations. Biosynthesis of magnetosome-like nanoparticles in mammalian cells would provide an endogenous magnetic resonance (MR) contrast agent under genetic control. This patented technology would provide long-term molecular imaging for tracking cellular and molecular activities throughout the cell\u27s life cycle