Jana Mihalic

Magnetic resonance imaging contrast of iron oxide nanoparticles developed for hyperthermia is dominated by iron content

Abstract Purpose: Magnetic iron oxide nanoparticles (MNPs) are used as contrast agents for magnetic resonance imaging (MRI) and hyperthermia for cancer treatment. The relationship between MRI signal intensity and cellular iron concentration for many new formulations, particularly MNPs having magnetic properties designed for heating in hyperthermia, is lacking. In this study, we examine the correlation between MRI T2 relaxation time and iron content in cancer cells loaded with various MNP formulations. Materials and methods: Human prostate carcinoma DU-145 cells were loaded with starch-coated bionised nanoferrite (BNF), iron oxide (Nanomag® D-SPIO), Feridex™, and dextran-coated Johns Hopkins University (JHU) particles at a target concentration of 50 pg Fe/cell using poly-D-lysine transfection reagent. T2-weighted MRI of serial dilutions of these labelled cells was performed at 9.4 T and iron content quantification was performed using inductively coupled plasma mass spectrometry (ICP-MS). Clonogenic assay was used to characterise cytotoxicity. Results: No cytotoxicity was observed at twice the target intracellular iron concentration (∼100 pg Fe/cell). ICP-MS revealed highest iron uptake efficiency with BNF and JHU particles, followed by Feridex and Nanomag-D-SPIO, respectively. Imaging data showed a linear correlation between increased intracellular iron concentration and decreased T2 times, with no apparent correlation among MNP magnetic properties. Conclusions: This study demonstrates that for the range of nanoparticle concentrations internalised by cancer cells the signal intensity of T2-weighted MRI correlates closely with absolute iron concentration associated with the cells. This correlation may benefit applications for cell-based cancer imaging and therapy including nanoparticle-mediated drug delivery and hyperthermia.

Preliminary study of injury from heating systemically delivered, nontargeted dextran- superparamagnetic iron oxide nanoparticles in mice

AIM To assess the potential for injury to normal tissues in mice due to heating systemically delivered magnetic nanoparticles in an alternating magnetic field (AMF). MATERIALS & METHODS Twenty three male nude mice received intravenous injections of dextran-superparamagnetic iron oxide nanoparticles on days 1-3. On day 6, they were exposed to AMF. On day 7, blood, liver and spleen were harvested and analyzed. RESULTS Iron deposits were detected in the liver and spleen. Mice that had received a high-particle dose and a high AMF experienced increased mortality, elevated liver enzymes and significant liver and spleen necrosis. Mice treated with low-dose superparamagnetic iron oxide nanoparticles and a low AMF survived, but had elevated enzyme levels and local necrosis in the spleen. CONCLUSION Magnetic nanoparticles producing only modest heat output can cause damage, and even death, when sequestered in sufficient concentrations. Dextran-superparamagnetic iron oxide nanoparticles are deposited in the liver and spleen, making these the sites of potential toxicity. Original submitted 16 August 2011; Revised submitted 21 March 2012; Published online 26 July 2012.

The effect of cell-cluster size on intracellular nanoparticle-mediated hyperthermia: is it possible to treat microscopic tumors?

AIM To compare the measured surface temperature of variable size ensembles of cells heated by intracellular magnetic fluid hyperthermia with heat diffusion model predictions. MATERIALS & METHODS Starch-coated Bionized NanoFerrite (Micromod Partikeltechnologie GmbH, Rostock, Germany) iron oxide magnetic nanoparticles were loaded into cultured DU145 prostate cancer cells. Cell pellets of variable size were treated with alternating magnetic fields. The surface temperature of the pellets was measured in situ and the associated cytotoxicity was determined by clonogenic survival assay. RESULTS & CONCLUSION For a given intracellular nanoparticle concentration, a critical minimum number of cells was required for cytotoxic hyperthermia. Above this threshold, cytotoxicity increased with increasing cell number. The measured surface temperatures were consistent with those predicted by a heat diffusion model that ignores intercellular thermal barriers. These results suggest a minimum tumor volume threshold of approximately 1 mm(3), below which nanoparticle-mediated heating is unlikely to be effective as the sole cytotoxic agent.

The Longitudinal Dependence of Black Carbon Concentration on Traffic Volume in an Urban Environment

Abstract The purpose of this study was to evaluate the effect of traffic volume on ambient black carbon (BC) concentration in an inner-city neighborhood “hot spot” while accounting for modifying effects of weather and time. Continuous monitoring was conducted for 12 months at the Baltimore Traffic Study site surrounded by major urban streets that together carry over 150,000 vehicles per day. Outdoor BC concentration was measured with an Aethalometer; vehicles were counted pneumatically on two nearby streets. Meteorological data were also obtained. Missing data were imputed and all data were normalized to a 5-min observational interval (n = 105,120). Time-series modeling accounted for autoregressively (AR) correlated errors. This study found that outdoor BC was positively correlated at a statistically significant level with neighborhood-level vehicle counts, which contributed at a rate of 66 ± 10 (SE) ng/m3 per 100 vehicles every 5 min. Winds from the SW-S-SE quarter were associated with the greatest increases in BC (376-612 ng/m3). These winds would have entrained BC from Baltimore’s densely trafficked central business district, as well as a nearby interstate highway. The strong influence of wind direction implicates atmospheric transport processes in determining BC exposure. Dew point, mixing height, wind speed, season, and workday were also statistically significant predictors. Background exposure to BC was estimated to be 905 ng/m3. The optimal, statistically significant representation of BC’s autocorrelation was AR([1:6]) × 288 × 2016, where the short-term AR factor (lags 1-6) indicated that BC concentrations are correlated for up to 30 min, and the AR factors for lags 288 and 2016 indicate longer-term autocorrelations at diurnal and weekly cycles, respectively. It was concluded that local exposure to BC from mobile sources is substantially modified by meteorological and temporal conditions, including atmospheric transport processes. BC concentration also demonstrates statistically significant autocorrelation at several time scales.

Assessment of heterogeneity of metal composition of fine particulate matter collected from eight U.S. counties using principal component analysis

The main objectives of this study are to (1) characterize chemical constituents of particulate matter (PM) and (2) compare overall differences in PM collected from eight U.S. counties. This project was undertaken as a part of a larger research program conducted by the Johns Hopkins Particulate Matter Research Center (JHPMRC). The goal of the JHPMRC is to explore the relationship between health effects and exposure to ambient PM of differing composition. The JHPMRC collected weekly filter-based ambient fine particle samples from eight U.S. counties between January 2008 and January 2010. Each sampling effort consisted of a 5–6-week sampling period. Filters were analyzed for 25 metals using inductively coupled plasma mass spectrometry (ICP-MS). Overall compositional differences were ranked by principal component analysis (PCA). The results showed that weekly concentrations of each element varied 3–40 times between the eight counties. PCA showed that the first five principal components explained 85% of the total variance. The authors found significant overall compositional differences in PM as the average of standardized principal component scores differed between the counties. These findings demonstrate PCA is a useful tool to identify the differences in PM compositional mixtures by county. These differences will be helpful for epidemiological and toxicological studies to help explain why health risks associated with PM exposure are different in locations with similar mass concentrations of PM. Implications: Previous studies have demonstrated associations between health effects and particulate matter (PM) using a single component or a combination of few components. Other studies have shown constituents of PM can vary greatly by location and that these differences may explain why the health effects associated with PM exposure are different by location. However, a single or a combination of a few components cannot represent PM as a whole. To address the need for evaluating PM as a complex mixture, the authors demonstrated the utility of principal component analysis to assess heterogeneity of PM.

Design and characterization of a sequential cyclone system for the collection of bulk particulate matter

In this paper, we describe the design, development and characterization of a high-volume sequential cyclone system for the collection of size-segregated PM in dry bulk form from the ambient environment in sufficient quantity for physical, chemical and toxicological characterization. The first stage of the system consists of a commercially available high volume PM(10) inlet. The second stage cyclone was designed by us to collect inhalable coarse particles (<10 µm and >2.5 µm). When tested individually with a challenge aerosol, a D(50) cut-size of this stage was found to be 2.3 µm at a flow rate of 1 m(3) min(-1). The third stage, a commercially available cyclone designed for surface dust sampling, had a D(50) cut-size of 0.3 µm when tested at the same flow rate. The purpose of the third stage is to collect the fine particle portion of PM(2.5) or accumulation mode (PM <2.5 µm and >0.1 µm). Thus, the sequential cyclone system will collect bulk samples of both the inhalable coarse particles and the fine particle portion of PM(2.5). The operation and maintenance of the new system are straightforward and allow for reliable collection of dry bulk ambient PM at relatively low cost.

Characterization of intratumor magnetic nanoparticle distribution and heating in a rat model of metastatic spine disease: Laboratory investigation

OBJECT The goal of this study was to optimize local delivery of magnetic nanoparticles in a rat model of metastatic breast cancer in the spine for tumor hyperthermia while minimizing systemic exposure. METHODS A syngeneic mammary adenocarcinoma was implanted into the L-6 vertebral body of 69 female Fischer rats. Suspensions of 100-nm starch-coated iron oxide magnetic nanoparticles (micromod Partikeltechnologie GmbH) were injected into tumors 9 or 13 days after implantation. For nanoparticle distribution studies, tissues were harvested from a cohort of 36 rats, and inductively coupled plasma mass spectrometry and histopathological studies with Prussian blue staining were used to analyze the samples. Intratumor heating was tested in 4 anesthetized animals with a 20-minute exposure to an alternating magnetic field (AMF) at a frequency of 150 kHz and an amplitude of 48 kA/m or 63.3 kA/m. Intratumor and rectal temperatures were measured, and functional assessments of AMF-exposed animals and histopathological studies of heated tumor samples were examined. Rectal temperatures alone were tested in a cohort of 29 rats during AMF exposure with or without nanoparticle administration. Animal studies were completed in accordance with the protocols of the University Animal Care and Use Committee. RESULTS Nanoparticles remained within the tumor mass within 3 hours of injection and migrated into the bone at 6, 12, and 24 hours. Subarachnoid accumulation of nanoparticles was noted at 48 hours. No evidence of lymphoreticular nanoparticle exposure was found on histological investigation or via inductively coupled plasma mass spectrometry. The mean intratumor temperatures were 43.2°C and 40.6°C on exposure to 63.3 kA/m and 48 kA/m, respectively, with histological evidence of necrosis. All animals were ambulatory at 24 hours after treatment with no evidence of neurological dysfunction. CONCLUSIONS Locally delivered magnetic nanoparticles activated by an AMF can generate hyperthermia in spinal tumors without accumulating in the lymphoreticular system and without damaging the spinal cord, thereby limiting neurological dysfunction and minimizing systemic exposure. Magnetic nanoparticle hyperthermia may be a viable option for palliative therapy of spinal tumors.

An optimised spectrophotometric assay for convenient and accurate quantitation of intracellular iron from iron oxide nanoparticles

Abstract We report the development and optimisation of an assay for quantitating iron from iron oxide nanoparticles in biological matrices by using ferene-s, a chromogenic compound. The method is accurate, reliable and can be performed with basic equipment common to many laboratories making it convenient and inexpensive. The assay we have developed is suited for quantitation of iron in cell culture studies with iron oxide nanoparticles, which tend to manifest low levels of iron. The assay was validated with standard reference materials and with inductively coupled plasma-mass spectrometry (ICP-MS) to accurately measure iron concentrations ∼1 × 10−6 g in about 1 × 106 cells (∼1 × 10−12 g Fe per cell). The assay requires preparation and use of a working solution to which samples can be directly added without further processing. After overnight incubation, the absorbance can be measured with a standard UV/Vis spectrophotometer to provide iron concentration. Alternatively, for expedited processing, samples can be digested with concentrated nitric acid before addition to the working solution. Optimization studies demonstrated significant deviations accompany variable digestion times, highlighting the importance to ensure complete iron ion liberation from the nanoparticle or sample matrix to avoid underestimating iron concentration. When performed correctly, this method yields reliable iron ion concentration measurements to ∼2 × 10−6 M (1 × 10−7 g/ml sample).

Physical characterization and in vivo organ distribution of coated iron oxide nanoparticles

Citrate-stabilized iron oxide magnetic nanoparticles (MNPs) were coated with one of carboxymethyl dextran (CM-dextran), polyethylene glycol-polyethylene imine (PEG-PEI), methoxy-PEG-phosphate+rutin, or dextran. They were characterized for size, zeta potential, hysteresis heating in an alternating magnetic field, dynamic magnetic susceptibility, and examined for their distribution in mouse organs following intravenous delivery. Except for PEG-PEI-coated nanoparticles, all coated nanoparticles had a negative zeta potential at physiological pH. Nanoparticle sizing by dynamic light scattering revealed an increased nanoparticle hydrodynamic diameter upon coating. Magnetic hysteresis heating changed little with coating; however, the larger particles demonstrated significant shifts of the peak of complex magnetic susceptibility to lower frequency. 48 hours following intravenous injection of nanoparticles, mice were sacrificed and tissues were collected to measure iron concentration. Iron deposition from nanoparticles possessing a negative surface potential was observed to have highest accumulation in livers and spleens. In contrast, iron deposition from positively charged PEG-PEI-coated nanoparticles was observed to have highest concentration in lungs. These preliminary results suggest a complex interplay between nanoparticle size and charge determines organ distribution of systemically-delivered iron oxide magnetic nanoparticles.

Immunodetection and quantification of airborne (1–3)-β-D-glucan-carrying particles with the halogen immunoassay

Fungal cell wall components, such as (1-3)-β-D-glucan, are known to be capable of activating the innate immune system and pose a respiratory health risk in different environments. Mass-based non-viable techniques commonly used for assessment of fungal exposures could be β-D-glucan-specific, but are limited to analysis of liquid extracts. The variable solubility of different β-D-glucans may underestimate β-D-glucan exposure and long sampling times required for mass-based methods make assessing short-term exposures difficult. In this study, we evaluated the utility of the halogen immunoassay (HIA), an immunoblotting technique previously used for allergens, to immunodetect and quantify β-D-glucan-carrying particles (BGCPs). The HIA was able to detect BGCPs without background staining when β-D-glucan standards and air samples collected at a poultry house during short sampling periods were evaluated. The image analysis protocol previously developed by our group for mouse allergen allowed simultaneous immunodetection and quantification of β-D-glucan-containing particles. Our results suggest that the HIA holds promise for quantifying β-D-glucan exposures. To our knowledge, this is the first time in which the HIA was used for non-allergenic compounds of microbial or fungal origins.