Microemulsion-made gadolinium carbonate hollow nanospheres showing magnetothermal heating and drug release.

Gadolinium carbonate (Gd2(CO3)3) hollow nanospheres and their suitability for drug transport and magnetothermally-induced drug release are presented. The hollow nanospheres are prepared via a microemulsion-based synthesis using tris(tetramethylcyclopentadienyl)gadolinium(iii) and CO2 as the starting materials. Size, structure and composition of the as-prepared Gd2(CO3)3 hollow nanospheres are comprehensively validated by several independent analytical methods (HRTEM, HAADF-STEM, DLS, EDXS, XRD, FT-IR, DTA-TG). Accordingly, they exhibit an outer diameter of 26 ± 4 nm, an inner cavity of 7 ± 2 nm, and a wall thickness of 9 ± 3 nm. As a conceptual study, the nanocontainer-functionality of the Gd2(CO3)3 hollow nanospheres is validated upon filling with the anti-cancerogenic agent doxorubicin (DOX), which is straightforward via the microemulsion (ME) strategy. The resulting DOX@Gd2(CO3)3 nanocontainers provide the option of multimodal imaging including optical and magnetic resonance imaging (OI, MRI) as well as magnetothermal heating and drug release. As a proof-of-concept, we could already prove the intrinsic DOX-based fluorescence, a low systemic toxicity according to in vitro studies as well as the magnetothermal effect and a magnetothermally-induced DOX release. In particular, the latter is new for Gd-containing nanoparticles and highly promising in view of theranostic nanocontainers and synergistic physical and chemical tumor treatment.

Improved Hyperthermia Treatment of Tumors Under Consideration of Magnetic Nanoparticle Distribution Using Micro-CT Imaging.

PURPOSE: Heterogeneous magnetic nanoparticle (MNP) distributions within tumors can cause regions of temperature under dosage and reduce the therapeutic efficiency. Here, micro-computed tomography (CT) imaging was used as a tool to determine the MNP distribution in vivo. The therapeutic success was evaluated based on tumor volume and temperature distribution. PROCEDURES: Tumor-bearing mice were intratumorally injected with iron oxide particles. MNP distribution was assessed by micro-CT with a low radiation dose protocol. RESULTS: MNPs were clearly visible, and the exact distribution to nontumor structures was detected by micro-CT. Knowledge of the intratumoral MNP distribution allowed the generation of higher temperatures within the tumor and led to higher temperature values after exposure to an alternating magnetic field (AMF). Consequently, the tumor size after 28 days was reduced to 14 and 73 % of the initial tumor volume for the MNP/AMF/CT and MNP/AMF groups, respectively. CONCLUSIONS: The MNP distribution pattern mainly governed the generated temperature spots in the tumor. Knowing the MNP distribution enabled individualized hyperthermia treatment and improved the overall therapeutic efficiency.

Magnetorelaxometry for localization and quantification of magnetic nanoparticles for thermal ablation studies.

In magnetic heating treatments, intratumorally injected superparamagnetic iron oxide nanoparticles (MNP) exposed to an externally applied alternating magnetic field generate heat, specifically at the tumor region. This inactivates cancer cells with minimal side effects to the normal tissue. Therefore, the quantity of MNP needs to be thoroughly controlled to govern adequate heat production. Here, we demonstrate the capability of magnetorelaxometry (MRX) for the non-invasive quantification and localization of MNP accumulation in small animal models. The results of our MRX measurements using a multichannel vector magnetometer system with 304 SQUIDs (superconductive quantum interference device) on three mice hosting different carcinoma models (9L/lacZ and MD-AMB-435) are presented. The position and magnitude of the magnetic moment are reconstructed from measured spatial magnetic field distributions by a magnetic dipole model fit applying a Levenberg-Marquadt algorithm. Therewith, the center of gravity and the total amount of MNP accumulation in the mice are determined. Additionally, for a fourth mouse the distribution of MNP over individual organs and the tumor is analyzed by single-channel SQUID measurements, obtaining a sensitive spatial quantification. This study shows that magnetorelaxometry is well suited to monitor MNP accumulation before cancer therapy, with magnetic heating being an important precondition for treatment success.

[Magnetically based enhancement of nanoparticle uptake in tumor cells: combination of magnetically induced cell labeling and magnetic heating]. / Magnetisch basierte Steigerung der Nanopartikelaufnahme in Tumorzellen: Kombination von magnetisch induzierter Zellmarkierung und magnetischer Wärmebehandlung.

PURPOSE: Magnetic nanoparticles (MNP) are known to be versatile tools in diagnostic and interventional radiology. The goal of the present study was to assess whether MNP can be selectively accumulated on human adenocarcinoma cells in vitro using an external magnetic field (magnetically induced cell labeling) and whether these labeled tumor cells can then be destroyed after being exposed to an alternating magnetic field (magnetically induced heating). In this context, a long-term goal is to combine these two developing methods to achieve an additive effect in tumor therapy. MATERIALS AND METHODS: BT-474 cells were incubated until confluence. Magnetic nanoparticles (0.32 mg Fe/ml culture medium) were then added and the flask was exposed to an external magnetic field gradient (magnetically induced cell labeling, 56 or 83 mT magnets) for 24 hours in order to label the tumor cells with nanoparticles. Cells without both MNP and magnetic labeling as well as cells with MNP incubation but without magnetic labeling served as controls. After MNP incubation, the magnetically labeled cells (5 x 10 (7) cells/ml) were exposed to an alternating magnetic field for 5.45 minutes (frequency 400 kHz, amplitude 24.6 kA/m). The combination effect of both magnetic labeling and magnetic heating was assessed by determining the temperature increase. The amount of MNP accumulated within the cells was determined by measuring the iron content via atomic absorption spectrometry. For statistical analysis mean values and standard deviations of temperature increases and iron contents were calculated and the differences were analyzed using the Student's t-test. RESULTS: A significant temperature increase (p < 0.01) during magnetic heating of 41.76 +/- 4.60 K was detected after magnetic labeling of the cells (5 x 10 (7) cells/ml, 83 mT) incubated with MNP. In comparison, the cells incubated with MNP but without magnetic labeling revealed a temperature increase of 32.03 +/- 3.33 K, naked cells of only 2.69 +/- 0.34 K. CONCLUSION: The results demonstrated the magnetically based enhancement of cellular uptake of nanoparticles by tumor cells, resulting in the intensification of the generated temperature increase during magnetic heating. Consequently, magnetic nanoparticles are shown to be valuable tools for the combination of magnetically based therapy modalities.

[Magnetic thermotherapy of breast tumors: an experimental therapeutic approach]. / Magnetische Thermotherapie von Tumoren der Brust: ein experimenteller Therapieansatz.

The therapeutic strategy for breast cancer is changing, especially for early tumor stages with good prognosis. One potential minimally invasive therapy modality consists in the accumulation of a well-tolerated magnetic material (iron oxides, particularly magnetite) in the target tissue. By applying an alternating magnetic field, energy is selectively absorbed and induces harmful heating of the tumor. The present review deals with the essential conditions and parameters as studied in vitro and in vivo in animal experiments. Extrapolations to the clinical situation are discussed, in particular, the heating potential of the magnetic material, the selection of the magnetic field parameters, the occurrence of eddy currents, the generation of localized heating spots and the expected temperature rises and their effects on the tumor area.

Use of magnetic nanoparticle heating in the treatment of breast cancer.

Magnetic nanoparticles are promising tools for the minimal invasive elimination of small tumours in the breast using magnetically-induced heating. The approach complies with the increasing demand for breast conserving therapies and has the advantage of offering a selective and refined tuning of the degree of energy deposition allowing an adequate temperature control at the target. The biophysical basis of the approach, the magnetic and structural properties of magnetic nanoparticles are reviewed. Results with model targets and in vivo experiments in laboratory animals are reported.

[In vitro imaging of magnetite particles]. / Bildgebende Darstellung von Magnetiten in vitro.

PURPOSE: To find an optimal imaging modality for the assessment of magnetite agglomerations used as the heating sources during magnetic thermoablation of tumors. METHODS: 1 to 107 mg of coated (starch) magnetite particles were directly administered to an in vitro tumor model (swine lymph nodes) and investigated immediately (radiography) or after being embedded within a 4 % agar-phantom (sonography). T1-weighted MR images (TR = 400 ms, TE = 14 ms) were acquired from lymph nodes containing 0.5 to 25 mg magnetite. RESULTS: All investigated magnetite masses were qualitatively detectable by radiography. Sonographically, only mass agglomerations containing 107 mg magnetite were appropriately discernible. MRT images revealed distinct susceptibility artifacts. CONCLUSIONS: Based on the investigated imaging modalities, radiography is the method of choice for assessment of magnetite agglomerations using relevant dosages for magnetic thermoablation of tumors.

Vacuum-assisted resection of malignant tumors with and without subsequent radiofrequency ablation: feasibility of complete tumor treatment tested in an animal model.

PURPOSE: To evaluate the feasibility of vacuum-assisted tumor excision with and without RF ablation for the minimally invasive treatment of small tumors. MATERIALS AND METHODS: Twenty VX2 tumors were implanted bilaterally into the spine muscle of 10 rabbits. Tumor excision was performed after tumor sizes reached 10 mm (12-27 d incubation) with use of a vacuum-assisted biopsy device. Three or four directed vacuum-assisted biopsies were performed in angle steps of 30 degrees. In 10 tumors, ultrasound (US)-guided radiofrequency (RF) ablation (8 min, 60 W) was subsequently performed with use of a cooled-tip electrode system. Follow-up US was performed at 3-4-day intervals for as long as 3 weeks after excision/RF ablation. Autopsy and histopathologic analysis were performed. RESULTS: The duration of vacuum excision ranged from 12 to 45 minutes (25 min +/- 7). Histologically tumor-free margins in the outer round of the core biopsy specimens were found in only four of 20 cases (20%). Maximum lesion sizes during RF ablation ranged from 18 to 25 mm (20 mm +/- 2.6). Histologic examination of the excision specimens documented tumor-free margins in only three tumors (30%) among the excision-only group and only one (10%) among the combined excision/ablation group. Local recurrences occurred in eight of 10 cases (80%) after vacuum excision alone, whereas recurrence after combined excision and RF ablation occurred only in two of 10 cases (20%; P <.05). CONCLUSIONS: Local tumor resection with use of vacuum-assisted biopsy is feasible and promising as a minimally invasive therapy for the treatment of small focal breast neoplasms. Combined excision and RF ablation techniques may reduce the rate of local recurrence considerably.

Electromagnetic heating of breast tumors in interventional radiology: in vitro and in vivo studies in human cadavers and mice.

PURPOSE: To assess relevant parameters for the minimally invasive elimination of breast tumors by using a selective application of magnetite and exposure of the breast to an alternating magnetic field. MATERIALS AND METHODS: The specific absorption rate (SAR) of different magnetite samples was determined calorimetrically. Temperature elevations based on magnetite mass (7-112 mg) and magnetic field amplitude (1.2-6.5 kA/m frequency, 400 kHz) were investigated by using human breast tissue. Parameter combinations (21 mg +/- 9 [SD], 242-second magnetic field exposure, 6.5-kA/m amplitude) were tested in 10 immunodeficient mice bearing human adenocarcinomas (MX-1 cells). Histologic sections of heated tumor tissue were analyzed. RESULTS: SAR data of different magnetite particle types ranged from 3 to 211 W/g. Temperature elevation (DeltaT) as a function of the magnetite mass increased linearly up to 28 mg; at higher masses, a saturation of DeltaT was observed at nearly 88 degrees C. The dependence of DeltaT on magnetic field amplitude (H) revealed a third-order power law: DeltaT = 0.26 degrees C/(kA/m)(3). H(3), with r(2) = 0.95. A mean temperature of 71 degrees C +/- 8 was recorded in the tumor region at the end of magnetic field exposure of the mice. Typical macroscopic findings included tumor shrinkage after heating. Histologically nuclear degenerations were observed in heated malignant cells. CONCLUSION: Magnetic heating of breast tumors is a promising technique for future interventional radiologic treatments.

Saline-enhanced radiofrequency ablation of breast tissue: an in vitro feasibility study.

RATIONALE AND OBJECTIVES: The feasibility of radiofrequency (RF) ablation for the treatment of breast tumors was investigated in vitro. The best parameters for ablation of breast tissue were chosen. METHODS: Saline-enhanced RF ablation was performed in human breast tissue specimens and cow udder tissue. Temperature profiles were measured depending on RF power (20, 28, 36 W) and NaCl infusion rate (15, 30, 60 mL/h) using eight thermocouples. Lesion development was monitored by ultrasound. Thermolysis efficiency was measured by tissue weight determinations before and after ablation. RESULTS: After RF ablation of tissue samples, 73.6% turned into a fat/saline emulsion. Ultrasound monitoring showed a cone-shaped hyperechoic area during the first 2 minutes of RF ablation, followed by an irregular expansion of the area. Time-dependent spatial temperature curves were more homogeneous at low infusion rates (15 mL/h). Peak temperatures up to 160 degrees C were measured. CONCLUSIONS: Controlled RF ablation of breast tissue is feasible. The irregular expansion of RF lesions in fatty breast tissue is due to liquefied fat. Low saline interstitial infusion rates result in better control of lesioning.

Effects of magnetic thermoablation in muscle tissue using iron oxide particles: an in vitro study.

RATIONALE AND OBJECTIVES: To study the effects of magnetic thermoablation in muscle tissue from cow to assess interrelations that might be relevant for a minimally invasive therapy system in the long term. METHODS: Magnetite particles (50-180 mg) were placed in muscle tissue. Temperature elevations as a function of time and distance from the center of the magnetite deposition area were measured during the exposure (up to 304 seconds) to an alternating magnetic field (frequency 400 kHz, amplitude 6.5 kA/m) generated by a circular coil (diameter 90 mm). Measured curves were reproduced by numerical calculations. Tissue alterations, macroscopically visible as light-brown discoloration, were recorded by volume estimations and histopathologic studies. RESULTS: Significant temperature elevations (up to 87 degrees C) were reported within a distance of less than 15 mm from the magnetite deposition area. High initial heating rates were observed during the first 150 seconds of heating. The reproduction of the measured curves by numerical calculations was good (SD = 0.7 degrees C). The theoretical simulation was verified and applied to situations beyond the range of experimental conditions. Damaged tissue comprised pyknotic cell nuclei and degenerated myofibrils. Corresponding volumes were found to be up to 10 times higher than the volume of iron oxide dispersion. CONCLUSIONS: The data demonstrate the applicability of local magnetic thermoablation for therapy of muscle lesions in the long term.

First experiences in using a new ultrasound mode and ultrasound contrast agent in the diagnosis of blunt renal trauma: a feasibility study in an animal model.

RATIONALE AND OBJECTIVES: To evaluate the diagnostic value of the new ultrasound mode "wide-band harmonic" (WBH) using an ultrasound contrast agent in blunt renal trauma in an animal model. METHODS: A defined blunt renal trauma was induced in 10 rabbits according to published standards. Ultrasound (B-mode, color and power Doppler, WBH) was performed before and after trauma, with and without using a contrast agent (Levovist). Ultrasound features were compared with histologic findings. RESULTS: In 2 of the 10 rabbits, three focal renal intraparenchymal lesions with diameters ranging from 1.0 to 1.8 mm were found that could be identified only using WBH with contrast. Six of the 10 rabbits developed a subcapsular hematoma with a thickness of up to 1.5 mm, which was identified by conventional B-mode as well as WBH. Histologic workup confirmed these findings of intraparenchymal hematomas and did not reveal further lesions. CONCLUSIONS: Only 20% of the experimental subjects developed parenchymal lesions with diameters of 1.0 mm or larger. All these lesions were identified only using WBH. These results indicate the potential to use WBH plus contrast for the diagnosis of blunt renal trauma.

Evaluation of temperature increase with different amounts of magnetite in liver tissue samples.

RATIONALE AND OBJECTIVES: The biologic effects of magnetically induced heating effects using iron oxide, magnetite, were examined in vitro in liver tissue samples as a first step toward potential applications in cancer therapy. METHODS: For the determination of the temperature profile around an iron oxide sample, a cylinder containing 170 mg of magnetite was constructed and placed into pureed liver tissue from pig, together with thermocouples of copper and constantan wires positioned at defined distances from it. Temperature measurements were performed during the exposure to an alternating magnetic field (frequency: 400 kHz; amplitude: approximately 6.5 kA/m) generated by a circular coil (90 mm of diameter). Moreover, variable amounts of magnetite (dissolved in approximately 0.2 mL physiologic saline) were injected directly into carrageenan gels. During the exposure to a magnetic field for 4 minutes the temperature increase was determined in the area of iron oxide deposition using a thermocouple. Additionally, variable amounts of magnetite were injected directly into isolated liver tissue samples (diameter: 20 mm; height: 30 mm) and exposed to a magnetic field for 2 minutes. The extent of the induced macroscopically visible tissue alterations (light brown colorations caused by heating) was examined by means of volume estimations. The degrees of cellular necrosis were investigated by histopathologic studies. RESULTS: The temperature profile around a magnetite cylinder revealed a significant decrease of temperature difference between the beginning and the end of heating, depending on increasing distance from the sample center. The extent of the temperature difference correlated with increasing heating time. No significant variations of temperature were observed at a distance of approximately 12 mm from the sample center. A good correlation (r = 0.98) between the injected amounts (31 to 200 mg) and the temperature increase since the start of heating (6.8-33.7 degrees C) in the area of iron oxide deposits was detected. The volume of damaged liver tissue was approximately seven times higher than the injected volume of iron oxide dispersion. Histologically different degrees of cellular necrosis were observed. CONCLUSIONS: The parameters determined in this article show that iron oxides are able to induce considerable heating effects in the surroundings. After an adequate optimization of the technical procedure, it is conceivable that heating properties of magnetites can be used in future cancer treatments.

Sensitivity of a hamster lung cell line to direct and indirect acting carcinogens.

Cytotoxicity of benzo(a)pyrene (B(a)P), 7,12-dimethylbenz(a)anthracene (DMBA), aflatoxin B1 (AB1), and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was estimated in vitro by using a hamster lung cell line. The studies were conducted to assess the usefulness of an organ-specific cell culture system for demonstrating differences in the cytotoxic potency of diverse chemical carcinogens. Cytotoxicity was determined by using the succinate dehydrogenase assay (MTT assay) after different incubation times and concentrations with the corresponding carcinogens. The effective concentration EC50 as well as the slope of the regression line were used as parameters for the biological effects. The results from these studies indicate a clear dose-dependent reaction after incubation of the cells with aflatoxin B1 (EC50: 2.3 microM) and MNNG (EC50: 4.0 microM). For the polycyclic hydrocarbons benzo(a)pyrene and DMBA, a dose-independent reaction was found. These results indicate that consideration of the EC50 values only might not be sufficient to characterize differences in the cytotoxic activity of different substances. Chemicals can lead to equal values in the EC50, but cells can differ significantly in their biological sensitivity, meaning that the extent of reduction in cell proliferation depends on the chemical used. By considering the two above-mentioned parameters, a ranking for the analyzed substances will be possible in the following way: AB1, MNNG, DMBA and B(a)P. Taken together, our experiments show that it is possible to reveal differences in the cytotoxic potency of chemicals by using in vitro methods.