Engineered Bacteria with Genetic Circuits Accumulating Nanomagnets as MRI Contrast Agents.

The demand for highly efficient cancer diagnostic tools increases alongside the high cancer incidence nowadays. Moreover, there is an imperative need for novel cancer treatment therapies that lack the side effects of conventional treatment options. Developments in this aspect employ magnetic nanoparticles (MNPs) for biomedical applications due to their stability, biocompatibility, and magnetic properties. Certain organisms, including many bacteria, can synthesize magnetic nanocrystals, which help their spatial orientation and survival by sensing the earth's geomagnetic field. This work aims to convert Escherichia coli to accumulate magnetite, which can further be coupled with drug delivery modules. The authors design magnetite accumulating bacterial machines using genetic circuitries hiring Mms6 with iron-binding activity and essential in magnetite crystal formation. The work demonstrates that the combinatorial effect of Mms6 with ferroxidase, iron transporter protein, and material binding peptide enhances the paramagnetic behavior of the cells in magnetic resonance imaging (MRI) measurements. Cellular machines are also engineered to display Mms6 peptide on the cell surface via an autotransporter protein that shows augmented MRI performance. The findings are promising for endowing a probiotic bacterium, able to accumulate magnetite intracellularly or extracellularly, serving as a theranostics agent for cancer diagnostics via MRI scanning and hyperthermia treatment.

Simultaneous temperature and viscosity estimation capability via magnetic nanoparticle relaxation.

PURPOSE: Magnetic particle imaging (MPI) is emerging as a highly promising imaging modality. Magnetic nanoparticles (MNPs) are used as imaging tracers in MPI, and their relaxation behavior provides the foundation for its functional imaging capability. Since MNPs are also utilized in magnetic fluid hyperthermia (MFH) and MPI enables localized MFH, temperature mapping arises as an important application area of MPI. To achieve accurate temperature estimations, however, one must also take into account the confounding effects of viscosity on the MPI signal. In this work, we analyze the effects of temperature and viscosity on MNP relaxation and determine temperature and viscosity sensitivities of relaxation time constant estimations via TAURUS (TAU estimation via Recovery of Underlying mirror Symmetry) at a wide range of operating points to empower simultaneous mapping of these two parameters. METHODS: A total of 15 samples were prepared to reach four target viscosity levels (0.9-3.6 mPa · $\cdot$ s) at five different temperatures (25-45 ∘ $^\circ$ C). Experiments were performed on a magnetic particle spectrometer (MPS) setup at 60 different operating points at drive field amplitudes ranging between 5 and 25 mT and frequencies ranging between 1 and 7 kHz. To enable these extensive experiments, an in-house arbitrary-waveform MPS setup with temperature-controlled heating capability was developed. The operating points were divided into four groups with comparable signal levels to maximize signal gain during rapid signal acquisition. The relaxation time constants were estimated via TAURUS, by restoring the underlying mirror symmetry property of the positive and negative half cycles of the time-domain MNP response. The relative time constants with respect to the drive field period, τ ̂ $\hat{\tau }$ , were computed to enable quantitative comparison across different operating points. At each operating point, a linear fit was performed to τ ̂ $\hat{\tau }$ as a function of each functional parameter (i.e., temperature or viscosity). The slopes of these linear fits were utilized to compute the temperature and viscosity sensitivities of TAURUS. RESULTS: Except for outlier behaviors at 1 kHz, the following global trends were observed: τ ̂ $\hat{\tau }$ decreases with drive field amplitude, increases with drive field frequency, decreases with temperature, and increases with viscosity. The temperature sensitivity varies slowly across the operating points and reaches a maximum value of 1.18%/ ∘ $^\circ$ C. In contrast, viscosity sensitivity is high at low frequencies around 1 kHz with a maximum value of 13.4%/(mPa · $\cdot$ s) but rapidly falls after 3 kHz. These results suggest that the simultaneous estimation of temperature and viscosity can be achieved by performing measurements at two different drive field settings that provide complementary temperature/viscosity sensitivities. Alternatively, temperature estimation alone can be achieved with a single measurement at drive field frequencies above 3 kHz, where viscosity sensitivity is minimized. CONCLUSIONS: This work demonstrates highly promising temperature and viscosity sensitivities for TAURUS, highlighting its potential for simultaneous estimation of these two environmental parameters via MNP relaxation. The findings of this work reveal the potential of a hybrid MPI-MFH system for real-time monitored and localized thermal ablation treatment of cancer.

Anisotropic dynamics of a self-assembled colloidal chain in an active bath.

Anisotropic macromolecules exposed to non-equilibrium (active) noise are very common in biological systems, and an accurate understanding of their anisotropic dynamics is therefore crucial. Here, we experimentally investigate the dynamics of isolated chains assembled from magnetic microparticles at a liquid-air interface and moving in an active bath consisting of motile E. coli bacteria. We investigate both the internal chain dynamics and the anisotropic center-of-mass dynamics through particle tracking. We find that both the internal and center-of-mass dynamics are greatly enhanced compared to the passive case, i.e., a system without bacteria, and that the center-of-mass diffusion coefficient D features a non-monotonic dependence as a function of the chain length. Furthermore, our results show that the relationship between the components of D parallel and perpendicular with respect to the direction of the applied magnetic field is preserved in the active bath compared to the passive case, with a higher diffusion in the parallel direction, in contrast to previous findings in the literature. We argue that this qualitative difference is due to subtle differences in the experimental geometry and conditions and the relative roles played by long-range hydrodynamic interactions and short-range collisions.

Calibration-Free Relaxation-Based Multi-Color Magnetic Particle Imaging.

Magnetic particle imaging (MPI) is a novel imaging modality with important potential applications, such as angiography, stem cell tracking, and cancer imaging. Recently, there have been efforts to increase the functionality of MPI via multi-color imaging methods that can distinguish the responses of different nanoparticles, or nanoparticles in different environmental conditions. The proposed techniques typically rely on extensive calibrations that capture the differences in the harmonic responses of the nanoparticles. In this paper, we propose a method to directly estimate the relaxation time constant of the nanoparticles from the MPI signal, which is then used to generate a multi-color relaxation map. The technique is based on the underlying mirror symmetry of the adiabatic MPI signal when the same region is scanned back and forth. We validate the proposed method via simulations, and via experiments on our in-house magnetic particle spectrometer setup at 10.8 kHz and our in-house MPI scanner at 9.7 kHz. Our results show that nanoparticles can be successfully distinguished with the proposed technique, without any calibration or prior knowledge about the nanoparticles.

Development of tailored SPION-PNIPAM nanoparticles by ATRP for dually responsive doxorubicin delivery and MR imaging.

Biocompatible, colloidally stable and ultra-small Fe3O4 nanoparticles (SPIONs) coated with poly(N-isopropylacrylamide) (PNIPAM) were synthesized via surface-initiated ATRP (atom transfer radical polymerization) to prevent excessive aggregation of magnetic cores and interparticle crosslinking, and to provide control over polymer content. These SPION-PNIPAM nanoparticles (NPs) have a hydrodynamic size between 8 and 60 nm depending on the PNIPAM content, and hence are ultrasmall in size and have an LCST around 38 °C. They had a high drug-loading capacity reaching 9.6 wt% doxorubicin in the final composition. The Dox release studies revealed pH and temperature-dependent release, which was not reported for PNIPAM before. Release of Dox under physiological conditions was below 20%, but around 90% at 42 °C and pH 5. This dually responsive nature is very advantageous to increase the drug efficacy and reduce side-effects, simultaneously. The cytocompatability of the SPION-PNIPAM NPs and the influence of Dox delivery to cells were investigated via in vitro cell viability, apoptosis, DNA-damage and confocal microscopy studies. The NPs were shown to be highly cytocompatible and induce significant cell death due to Dox when loaded with the drug. Besides, it was seen that the polymeric content can be used as an additional factor in tuning the release kinetics. Lastly, these nanoparticles reduced the signal intensity significantly in the T2 mode, acting as a potential SPION-based contrast agent. Overall, here, we demonstrate the design of small, smart theranostic nanoparticles with high drug-loading capacity and pH-dependent temperature-sensitive release characteristics with the ability to generate contrast in MRI.

A new class of cubic SPIONs as a dual-mode T1 and T2 contrast agent for MRI.

Superparamagnetic iron oxide nanoparticles (SPIONs) are widely used as a robust negative contrast agent on conventional MRI. In this study, we (a) synthesized a new class of cubic SPIONs as a dual-mode contrast agent in MRI and (b) showed the in-vivo feasibility of these nanaoparticles as a simultaneous positive and negative contrast agent. Relaxation properties and contrast enhancement analysis of the synthesized SPIONs with two different shapes (cubic vs. spherical) and three different sizes 7nm, 11nm, and 14nm were investigated to evaluate contrast enhancement in-vitro. In-vivo MRI experiments were performed on a 3T MR scanner, where a healthy anesthetized rat was imaged before, and from 20 to 80min after intravenous injection of 1mg/kg of contrast agent. Representative transmission electron microscopy (TEM) images of the synthesized nanoparticles reveal that the particles are well dispersed in a solvent and do not aggregate. The in-vitro relaxivity and contrast enhancement analysis show that, among all six SPIONs tested, 11-nm cubic SPIONs possess optimal molar relaxivities and contrast enhancement values, which can shorten the spin-lattice and spin-spin relaxation times, simultaneously. No noticeable toxicity is observed during in-vitro cytotoxicity analysis. In-vivo T1-and T2-weighted acquisitions at 60-min post-injection of 11-nm cubic SPIONs result in 64% and 48% contrast enhancement on the T1-and T2-weighted images, respectively. By controlling the shape and size of SPIONs, we have introduced a new class of cubic SPIONs as a synergistic (dual-mode) MRI contrast agent. 11-nm cubic SPIONs with smaller size and high positive and negative contrast enhancements were selected as a promising candidate for dual-mode contrast agent. Our proof-of-concept MRI experiments on rat demonstrate the in-vivo dual-mode contrast enhancement feasibility of these nanoparticles.