The Mark Coventry Award: PhotothermAA Gel Combined With Debridement, Antibiotics, and Implant Retention Significantly Decreases Implant Biofilm Burden and Soft-Tissue Infection in a Rabbit Model of Knee Periprosthetic Joint Infection.

BACKGROUND: Chronic periprosthetic joint infection (PJI) is a major complication of total joint arthroplasty. The underlying pathogenesis often involves the formation of bacterial biofilm that protects the pathogen from both host immune responses and antibiotics. The gold standard treatment requires implant removal, a procedure that carries associated morbidity and mortality risks. Strategies to preserve the implant while treating PJI are desperately needed. Our group has developed an anti-biofilm treatment, PhotothermAA gel, which has shown complete eradication of 2-week-old mature biofilm in vitro. In this study, we tested the anti-biofilm efficacy and safety of PhotothermAA in vivo when combined with debridement, antibiotics and implant retention (DAIR) in a rabbit model of knee PJI. METHODS: New Zealand white rabbits (n = 21) underwent knee joint arthrotomy, titanium tibial implant insertion, and inoculation with Xen36 (bioluminescent Staphylococcus aureus) after capsule closure. At 2 weeks, rabbits underwent sham surgery (n = 6), DAIR (n = 6), or PhotothermAA with DAIR (n = 9) and were sacrificed 2 weeks later to measure implant biofilm burden, soft-tissue infection, and tissue necrosis. RESULTS: The combination of anti-biofilm PhotothermAA with DAIR significantly decreased implant biofilm coverage via scanning electron microscopy compared to DAIR alone (1.8 versus 81.0%; P < .0001). Periprosthetic soft-tissue cultures were significantly decreased in the PhotothermAA with DAIR treatment group (log reduction: Sham 1.6, DAIR 2.0, combination 5.6; P < .0001). Treatment-associated necrosis was absent via gross histology of tissue adjacent to the treatment area (P = .715). CONCLUSIONS: The addition of an anti-biofilm solution like PhotothermAA as a supplement to current treatments that allow implant retention may prove useful in PJI treatment.

Combination d-Amino Acid and Photothermal Hydrogel for the Treatment of Prosthetic Joint Infections.

Prosthetic joint infection (PJI) is a devastating complication requiring surgical intervention and prolonged antimicrobial treatment. The prevalence of PJI is on the rise, with an average incidence of 60,000 cases per year and a projected annual cost of $1.85 billion in the US. The underlying pathogenesis of PJI involves the formation of bacterial biofilms that protect the pathogen from the host immune response and antibiotics, making it difficult to eradicate such infections. Biofilms on implants are also resistant to mechanical brushing/scrubbing methods of removal. Since the removal of biofilms is currently only achievable by the replacement of the prosthesis, therapies aimed at eradicating biofilms while enabling retention of implants will revolutionize the management of PJIs. To address severe complications associated with biofilm-related infections on implants, we have developed a combination treatment that is based on a hydrogel nanocomposite system, containing d-amino acids (d-AAs) and gold nanorods, which can be delivered and transforms from a solution to a gel state at physiological temperature for sustained release of d-AAs and light-activated thermal treatment of infected sites. Using this two-step approach to utilize a near-infrared light-activated hydrogel nanocomposite system for thermal treatment, following initial disruption with d-AAs, we were able to successfully demonstrate in vitro the total eradication of mature Staphylococcus aureus biofilms grown on three-dimensional printed Ti-6Al-4V alloy implants. Using a combination of cell assays, computer-aided scanning electron microscopy analyses, and confocal microscopy imaging of the biofilm matrix, we could show 100% eradication of the biofilms using our combination treatment. In contrast, we were only able to see 25% eradication of the biofilms using the debridement, antibiotics, and implant retention method. Moreover, our hydrogel nanocomposite-based treatment approach is adaptable in the clinical setting and capable of combating chronic infections brought about by biofilms on medical implants.

Standardized quantification of biofilm in a novel rabbit model of periprosthetic joint infection.

Periprosthetic joint infection (PJI) is one of the most devastating complications of total joint arthroplasty. The underlying pathogenesis involves the formation of bacterial biofilm that protects the pathogen from the host immune response and antibiotics, making eradication difficult. The aim of this study was to develop a rabbit model of knee PJI that would allow reliable biofilm quantification and permit the study of treatments for PJI. In this work, New Zealand white rabbits ( n = 19 ) underwent knee joint arthrotomy, titanium tibial implant insertion, and inoculation with Xen36 (bioluminescent Staphylococcus aureus) or a saline control after capsule closure. Biofilm was quantified via scanning electron microscopy (SEM) of the tibial explant 14 d after inoculation ( n = 3 noninfected, n = 2 infected). Rabbits underwent debridement, antibiotics, and implant retention (DAIR) ( n = 6 ) or sham surgery ( n = 2 noninfected, n = 6 infected) 14 d after inoculation, and they were sacrificed 14 d post-treatment. Tibial explant and periprosthetic tissues were examined for infection. Laboratory assays supported bacterial infection in infected animals. No differences in weight or C-reactive protein (CRP) were detected after DAIR compared to sham treatment. Biofilm coverage was significantly decreased with DAIR treatment when compared with sham treatment (61.4 % vs. 90.1 %, p < 0 .0011) and was absent in noninfected control explants. In summary, we have developed an experimental rabbit hemiarthroplasty knee PJI model with bacterial infection that reliably produces quantifiable biofilm and provides an opportunity to introduce treatments at 14 d. This model may be used to better understand the pathogenesis of this condition and to measure treatment strategies for PJI.

Hyperthermia-mediated changes in the tumor immune microenvironment using iron oxide nanoparticles.

Iron oxide nanoparticles (IONPs) have often been investigated for tumor hyperthermia. IONPs act as heating foci in the presence of an alternating magnetic field (AMF). It has been shown that hyperthermia can significantly alter the tumor immune microenvironment. Typically, mild hyperthermia invokes morphological changes within the tumor, which elicits a secretion of inflammatory cytokines and tumor neoantigens. Here, we focused on the direct effect of IONP-induced hyperthermia on the various tumor-resident immune cell subpopulations. We compared direct intratumoral injection to systemic administration of IONPs followed by application of an external AMF. We used the orthotopic 4T1 mouse model, which represents aggressive and metastatic breast cancer with a highly immunosuppressive microenvironment. A non-inflamed and 'cold' microenvironment inhibits peripheral effector lymphocytes from effectively trafficking into the tumor. Using intratumoral or systemic injection, IONP-induced hyperthermia achieved a significant reduction of all the immune cell subpopulations in the tumor. However, the systemic delivery approach achieved superior outcomes, resulting in substantial reductions in the populations of both innate and adaptive immune cells. Upon depletion of the existing dysfunctional tumor-resident immune cells, subsequent treatment with clinically approved immune checkpoint inhibitors encouraged the repopulation of the tumor with 'fresh' infiltrating innate and adaptive immune cells, resulting in a significant decrease of the tumor cell population.

Di- and tri-component spinel ferrite nanocubes: synthesis and their comparative characterization for theranostic applications.

Spinel ferrite nanocubes (NCs), consisting of pure iron oxide or mixed ferrites, are largely acknowledged for their outstanding performance in magnetic hyperthermia treatment (MHT) or magnetic resonance imaging (MRI) applications while their magnetic particle imaging (MPI) properties, particularly for this peculiar shape different from the conventional spherical nanoparticles (NPs), are relatively less investigated. In this work, we report on a non-hydrolytic synthesis approach to prepare mixed transition metal ferrite NCs. A series of NCs of mixed zinc-cobalt-ferrite were prepared and their magnetic theranostic properties were compared to those of cobalt ferrite or zinc ferrite NCs of similar sizes. For each of the nanomaterials, the synthesis parameters were adjusted to obtain NCs in the size range from 8 up to 15 nm. The chemical and structural nature of the different NCs was correlated to their magnetic properties. In particular, to evaluate magnetic losses, we compared the data obtained from calorimetric measurements to the data measured by dynamic magnetic hysteresis obtained under alternating magnetic field (AMF) excitation. Cobalt-ferrite and zinc-cobalt ferrite NCs showed high specific adsorption rate (SAR) values in aqueous solutions but their heating ability was drastically suppressed once in viscous media even for NCs as small as 12 nm. On the other hand, non-stoichiometric zinc-ferrite NCs showed significant but lower SAR values than the other ferrites, but these zinc-ferrite NCs preserved almost unaltered their heating trend in viscous environments. Also, the presence of zinc in the crystal lattice of zinc-cobalt ferrite NCs showed increased contrast enhancement for MRI with the highest T2 relaxation time and in the MPI signal with the best point spread function and signal-to-noise ratio in comparison to the analogue cobalt-ferrite NC. Among the different compositions investigated, non-stoichiometric zinc-ferrite NCs can be considered the most promising material as a multifunctional theranostic platform for MHT, MPI and MRI regardless of the media viscosity in which they will be applied, while ensuring the best biocompatibility with respect to the cobalt ferrite NCs.

Engineered magnetic nanoparticles enhance chlorophyll content and growth of barley through the induction of photosystem genes.

This study investigates the impact of an engineered magnetic nanoparticle (MNP) on a crop plant. For this purpose, a sonochemical synthetic approach was utilized in order to dope magnetic elements (Co and Nd) into technologically important iron oxide NPs. After being characterized by using TEM, SEM, and XRD instruments, the MNPs were hydroponically applied to barley plants with varying doses (from 125 to 1000 mg/L) both in germination (4 days) and early growing stages (3 weeks). Physiological responses, as well as expression of photosystem marker genes, were assessed. Compared to the untreated control, MNP treatment enhanced germination rate (~ 31%), tissue growth (8% in roots, 16% in shoots), biomass (~ 21%), and chlorophyll (a, b) (~ 20%), and carotenoids (~ 22%) pigments. In general, plants showed the highest growth enhancement at 125 or 250 mg/L treatment. However, higher doses diminished the growth indices. Compared to the control, the catalase activity was significantly reduced in the leaves (~ 33%, p < 0.005) but stimulated in the roots (~ 46%, p < 0.005). All tested photosystem marker genes (BCA, psbA, and psaA) were overexpressed in MNP-treated leaves than non-treated control. Moreover, the gene expressions were found to be proportionally increased with increasing MNP doses, indicating a positive correlation between MNPs and the photosynthetic machinery, which could contribute to the enhancement of plant growth.

Uncovering the Magnetic Particle Imaging and Magnetic Resonance Imaging Features of Iron Oxide Nanocube Clusters.

Multifunctional imaging nanoprobes continue to garner strong interest for their great potential in the detection and monitoring of cancer. In this study, we investigate a series of spatially arranged iron oxide nanocube-based clusters (i.e., chain-like dimer/trimer, centrosymmetric clusters, and enzymatically cleavable two-dimensional clusters) as magnetic particle imaging and magnetic resonance imaging probes. Our findings demonstrate that the short nanocube chain assemblies exhibit remarkable magnetic particle imaging signal enhancement with respect to the individually dispersed or the centrosymmetric cluster analogues. This result can be attributed to the beneficial uniaxial magnetic dipolar coupling occurring in the chain-like nanocube assembly. Moreover, we could effectively synthesize enzymatically cleavable two-dimensional nanocube clusters, which upon exposure to a lytic enzyme, exhibit a progressive increase in magnetic particle imaging signal at well-defined incubation time points. The increase in magnetic particle imaging signal can be used to trace the disassembly of the large planar clusters into smaller nanocube chains by enzymatic polymer degradation. These studies demonstrate that chain-like assemblies of iron oxide nanocubes offer the best spatial arrangement to improve magnetic particle imaging signals. In addition, the nanocube clusters synthesized in this study also show remarkable transverse magnetic resonance imaging relaxation signals. These nanoprobes, previously showcased for their outstanding heat performance in magnetic hyperthermia applications, have great potential as dual imaging probes and could be employed to improve the tumor thermo-therapeutic efficacy, while offering a readable magnetic signal for image mapping of material disassemblies at tumor sites.

Exploring the chelation-based plant strategy for iron oxide nanoparticle uptake in garden cress (Lepidium sativum) using magnetic particle spectrometry.

Although iron is one of Earth's most abundant elements, its availability to plants remains an agricultural challenge, particularly in high pH environments. At high pH, iron forms insoluble ferric oxide-hydroxides that makes it inaccessible to plants. It is estimated that 30% of the world's cropland is too alkaline for optimal plant growth. Staple crops, like rice, are particularly susceptible to iron deficiency, thereby, necessitating the need for continued research in developing iron-based fertilizers. Recent studies have demonstrated the potential of using iron oxide nanoparticles (IONPs) as fertilizers to address iron deficiency in plants, but some studies have generated conflicting results. One of the major challenges associated in investigating IONP plant uptake and translocation is the inability to distinguish between intact IONPs versus leached iron ions. In this study, we utilized a new approach based on magnetic particle spectrometry (MPS) to monitor the uptake and distribution of different sized (10 and 20 nm) chelated IONPs in plants. We exposed garden cress (Lepidium sativum) plants to EDTA-capped IONPs and observed an 8-fold enhancement in total biomass and 1.4 times increase in chlorophyll production compared to plants treated with a commercial chelated iron fertilizer (Fe-EDTA). Moreover, we demonstrated that the uptake and tissue distribution of IONPs can be quantitatively monitored using MPS, and the results of the analysis were validated by atomic absorption spectroscopy, which is the conventional method used to study IONP plant uptake. Our study demonstrates that MPS is a reliable, sensitive, and effective analytical tool for the development of IONP-based fertilizers.

Applications and challenges of using 3D printed implants for the treatment of birth defects.

Pediatric implants are a special subclass of a vast number of clinically used medical implants, uniquely designed to address the needs of young patients who are at the onset of their developmental growth stage. Given the vulnerability of the implant receiver, it is crucial that the implants manufactured for small children with birth-associated defects be given careful considerations and great attention to design detail to avoid postoperative complications. In this review, we focus on the most common types of medical implants manufactured for the treatment of birth defects originating from both genetic and environmental causes. Particular emphasis is devoted toward identifying the implant material of choice and manufacturing approaches for the fabrication of pediatric prostheses. Along this line, the emerging role of 3D printing to enable customized implants for infants with congenital disorders is presented, as well as the possible complications associated with prosthetic-related infections that is prevalent in using artificial implants for the treatment of birth malformations.

A novel synthetic route for high-index faceted iron oxide concave nanocubes with high T2 relaxivity for in vivo MRI applications.

Iron oxide nanoparticles (IONPs) with high-index facets have shown great potential as high performance T2 contrast agents for MRI. Previous synthetic approaches focused mainly on ion-directed or oxidative etching methods. Herein, we report a new synthetic route for preparing high-index faceted iron oxide concave nanocubes using a bulky coordinating solvent. Through the systematic replacement of a non-coordinating solvent, 1-octadecene, with trioctylamine, the solvent interaction with the nanoparticle surface is modified, thereby, promoting the growth evolution of the IONPs from spherical to concave cubic morphology. The presence of the bulky trioctylamine solvent results in particle size increase and the formation of nanoparticles with enhanced shape anisotropy. A well-defined concave nanocube structure was evident from the early stages of particle growth, further confirming the important role of bulky coordinating solvents in nanoparticle structural development. The unique concave nanocube morphology has a direct influence on the magnetic properties of the IONPs, ultimately leading to an ultra-high T2 relaxivity (862.2 mM-1 s-1), and a 2-fold enhancement in T2*-weighted in vivo MRI contrast compared to spherical IONP analogs.

Magnetic Glycol Chitin-Based Hydrogel Nanocomposite for Combined Thermal and d-Amino-Acid-Assisted Biofilm Disruption.

Bacterial biofilms are highly antibiotic resistant microbial cell associations that lead to chronic infections. Unlike free-floating planktonic bacterial cells, the biofilms are encapsulated in a hardly penetrable extracellular polymeric matrix and, thus, demand innovative approaches for treatment. Recent advancements on the development of gel-nanocomposite systems with tailored therapeutic properties provide promising routes to develop novel antimicrobial agents that can be designed to disrupt and completely eradicate preformed biofilms. In our study, we developed a unique thermoresponsive magnetic glycol chitin-based nanocomposite containing d-amino acids and iron oxide nanoparticles, which can be delivered and undergoes transformation from a solution to a gel state at physiological temperature for sustained release of d-amino acids and magnetic field actuated thermal treatment of targeted infection sites. The d-amino acids in the hydrogel nanocomposite have been previously reported to inhibit biofilm formation and also disrupt existing biofilms. In addition, loading the hydrogel nanocomposite with magnetic nanoparticles allows for combination thermal treatment following magnetic field (magnetic hyperthermia) stimulation. Using this novel two-step approach to utilize an externally actuated gel-nanocomposite system for thermal treatment, following initial disruption with d-amino acids, we were able to demonstrate in vitro the total eradication of Staphylococcus aureus biofilms, which were resistant to conventional antibiotics and were not completely eradicated by separate d-amino acid or magnetic hyperthermia treatments.

Sensitivity Limits for in vivo ELISA Measurements of Molecular Biomarker Concentrations.

The extremely high sensitivity that has been suggested for magnetic particle imaging has its roots in the unique signal produced by the nanoparticles at the frequencies of the harmonics of the drive field. That sensitivity should be translatable to other methods that utilize magnetic nanoparticle probes, specifically towards magnetic nanoparticle spectroscopy that is used to measure molecular biomarker concentrations for an "in vivo ELISA" assay approach. In this paper, we translate the predicted sensitivity of magnetic particle imaging into a projected sensitivity limit for in vivo ELISA. The simplifying assumptions adopted are: 1) the limiting noise in the detection system is equivalent to the minimum detectable mass of nanoparticles; 2) the nanoparticle's signal arising from Brownian relaxation is completely eliminated by the molecular binding event, which can be accomplished by binding the nanoparticle to something so massive that it can no longer physically rotate and is large enough that Neel relaxation is minimal. Given these assumptions, the equation for the minimum concentration of molecular biomarker we should be able to detect is obtained and the in vivo sensitivity is estimated to be in the attomolar to zeptomolar range. Spectrometer design and nonspecific binding are the technical limitations that need to be overcome to achieve the theoretical limit presented.

High-performance iron oxide nanoparticles for magnetic particle imaging - guided hyperthermia (hMPI).

Magnetic particle imaging (MPI) is an emerging imaging modality that allows the direct and quantitative mapping of iron oxide nanoparticles. In MPI, the development of tailored iron oxide nanoparticle tracers is paramount to achieving high sensitivity and good spatial resolution. To date, most MPI tracers being developed for potential clinical applications are based on spherical undoped magnetite nanoparticles. For the first time, we report on the systematic investigation of the effects of changes in chemical composition and shape anisotropy on the MPI performance of iron oxide nanoparticle tracers. We observed a 2-fold enhancement in MPI signal through selective doping of magnetite nanoparticles with zinc. Moreover, we demonstrated focused magnetic hyperthermia heating by adapting the field gradient used in MPI. By saturating the iron oxide nanoparticles outside of a field free region (FFR) with an external static field, we can selectively heat a target region in our test sample. By comparing zinc-doped magnetite cubic nanoparticles with undoped spherical nanoparticles, we could show a 5-fold improvement in the specific absorption rate (SAR) in magnetic hyperthermia while providing good MPI signal, thereby demonstrating the potential for high-performance focused hyperthermia therapy through an MPI-guided approach (hMPI).

Magnetic Nanoparticles: Material Engineering and Emerging Applications in Lithography and Biomedicine.

We present an interdisciplinary overview of material engineering and emerging applications of iron oxide nanoparticles. We discuss material engineering of nanoparticles in the broadest sense, emphasizing size and shape control, large-area self-assembly, composite/hybrid structures, and surface engineering. This is followed by a discussion of several non-traditional, emerging applications of iron oxide nanoparticles, including nanoparticle lithography, magnetic particle imaging, magnetic guided drug delivery, and positive contrast agents for magnetic resonance imaging. We conclude with a succinct discussion of the pharmacokinetics pathways of iron oxide nanoparticles in the human body -- an important and required practical consideration for any in vivo biomedical application, followed by a brief outlook of the field.

Iron Oxide and Titanium Dioxide Nanoparticle Effects on Plant Performance and Root Associated Microbes.

In this study, we investigated the effect of positively and negatively charged Fe3O4 and TiO2 nanoparticles (NPs) on the growth of soybean plants (Glycine max.) and their root associated soil microbes. Soybean plants were grown in a greenhouse for six weeks after application of different amounts of NPs, and plant growth and nutrient content were examined. Roots were analyzed for colonization by arbuscular mycorrhizal (AM) fungi and nodule-forming nitrogen fixing bacteria using DNA-based techniques. We found that plant growth was significantly lower with the application of TiO2 as compared to Fe3O4 NPs. The leaf carbon was also marginally significant lower in plants treated with TiO2 NPs; however, leaf phosphorus was reduced in plants treated with Fe3O4. We found no effects of NP type, concentration, or charge on the community structure of either rhizobia or AM fungi colonizing plant roots. However, the charge of the Fe3O4 NPs affected both colonization of the root system by rhizobia as well as leaf phosphorus content. Our results indicate that the type of NP can affect plant growth and nutrient content in an agriculturally important crop species, and that the charge of these particles influences the colonization of the root system by nitrogen-fixing bacteria.

Magnetic Particle Imaging Tracers: State-of-the-Art and Future Directions.

Magnetic particle imaging (MPI) is an emerging imaging modality with promising applications in diagnostic imaging and guided therapy. The image quality in MPI is strongly dependent on the nature of its iron oxide nanoparticle-based tracers. The selection of potential MPI tracers is currently limited, and the underlying physics of tracer response is not yet fully understood. An in-depth understanding of the magnetic relaxation processes that govern MPI tracers, gained through concerted theoretical and experimental work, is crucial to the development of optimized MPI tracers. Although tailored tracers will lead to improvements in image quality, tailored relaxation may also be exploited for biomedical applications or more flexible image contrast, as in the recent demonstration of color MPI.

Highly efficient antibacterial iron oxide@carbon nanochains from wüstite precursor nanoparticles.

A new hydrothermal synthesis approach involving the carbonization of glucose in the presence of wüstite (FeO) nanoparticles is presented, which leads to the fabrication of rapidly acting and potent antibacterial agents based on iron oxide@carbon (IO@C) nanochains. By using nonmagnetic FeO precursor nanoparticles that slowly oxidize into the magnetic Fe3O4 crystal structure under hydrothermal conditions, we were able to prepare well-defined and short-length IO@C nanochains that are highly dispersed in aqueous media and readily interact with bacterial cells, leading to a complete loss in bacterial cell viability within short incubation times at minimal dosage. The smaller IO@C nanochains synthesized using the FeO precursor nanoparticles can reach above 2-fold enhancement in microbe-killing activity when compared to the larger nanochains fabricated directly from Fe3O4 nanoparticles. In addition, the synthesized IO@C nanochains can be easily isolated using an external magnet and be subsequently recycled to effectively eradicate Escherichia coli cells even after five separate treatment cycles.

Effect of metal precursor on the growth and electrochemical sensing properties of Pt-Ag nanoboxes.

Pt-Ag nanoboxes prepared using a Pt(4+) versus Pt(2+) precursor show not only stoichiometric, but also catalytic effects on the nanobox growth, resulting in metal content and surface morphology differences. Electrochemical studies on these nanoboxes reveal the combined importance of composition and hollow, rough-walled nanostructure for improved detection of electroactive molecules.

Iron oxide-loaded hollow mesoporous silica nanocapsules for controlled drug release and hyperthermia.

Magnetic field-responsive iron oxide-loaded hollow mesoporous silica nanocapsules that exhibit high drug loading capacity were synthesized using polymer nanospheres as sacrificial templates. Due to their magnetic field induced heating and remotely triggered drug release capabilities, these hybrid nanomaterials provide an excellent platform for the combination of hyperthermia and chemotherapy.