Ingrid Hilger

Physical limits of hyperthermia using magnetite fine particles

Structural and magnetic properties of fine particles of magnetite are investigated with respect to the application for hyperthermia. Magnetic hysteresis losses are measured in dependence on the field amplitude for selected commercial powders and are discussed in terms of grain size and structure of the particles. For ferromagnetic powders as well as for ferrofluids, results of heating experiments within organic gels in a magnetic high frequency field are reported. The heating effect depends strongly on the magnetic properties of the magnetite particles which may vary appreciably for different samples in dependence on the particle size and microstructure. In particular, the transition from ferromagnetic to superparamagnetic behavior causes changes of the loss mechanism, and accordingly, of the heating effect. The maximum attainable heating effect is discussed in terms of common theoretical models. Rise of temperature at the surface of a small heated sample as well as in its immediate neighborhood in the surrounding medium is measured in dependence on time and is compared with solutions of the corresponding heat conductivity problem. Conclusions with respect to clinical applications are given.

Application of magnetite ferrofluids for hyperthermia

A comparative study is presented for the specific loss power generated by an external magnetic field in superparamagnetic as well as ferromagnetic magnetite particles suspended in molten and solidified gel. The field amplitude dependence of magnetic losses obeys power laws of third order for ferromagnetic samples and second order for superparamagnetic samples, respectively. Calorimetrically determined data are compared with results of hysteresis measurements. Consequences for the application for hyperthermia are discussed.

Temperature distribution as function of time around a small spherical heat source of local magnetic hyperthermia

A spherical region containing magnetic particles embedded in extended muscle tissue is taken as model of small breast carcinomas. Using analytically derived equations the spatial temperature distribution is calculated as function of the time for exposing to an alternating magnetic field. In vitro measurements with muscle tissue yielded such an agreement with the calculations that treatment of small tumors in slightly vascularized tissues on the base of mathematical predictions seems now to be more promising than in the past.

Heating potential of iron oxides for therapeutic purposes in interventional radiology.

RATIONALE AND OBJECTIVES In addition to their diagnostic applications, iron oxides could be used therapeutically to eliminate tumors with heat if their heating powers are adequate. The authors therefore examined the specific absorption rate (SAR) of different iron oxide (magnetite) samples suspended in water and in liquid or solidified gel. MATERIALS AND METHODS The authors compared two ferromagnetic fine powders (total particle size, >350 nm and 100 nm), five superparamagnetic ferrofluidic samples (total particle size, 10-280 nm), and a commercially available contrast medium (ferumoxides injectable solution, Endorem). The SARs of the magnetic material-suspended in distilled water or in liquid or solid agar-were estimated from time-dependent calorimetric measurements during exposure to an alternating current magnetic field (amplitude, 6.5 kA/m; frequency, 400 kHz). RESULTS SARs varied considerably between the different iron oxide samples. The highest value was found for a ferrofluidic sample (>93 W/g), while Endorem had little heating power (<0.1 W/g). The SAR was clearly dependent on the aggregation state of the matrix only for the large-particle-size ferromagnetic sample, yielding the highest values for particle suspensions in water (74 W/g) and lowest for solid agar (8 W/g). The heating power of the smaller-particle-size ferromagnetic sample did not exceed 8 W/g. CONCLUSION Heating powers differed according to the interaction of multiple physical parameters. Iron oxides should be selected carefully for therapeutic applications in magnetic heating.

Magnetic multicore nanoparticles for hyperthermia--influence of particle immobilization in tumour tissue on magnetic properties.

When using magnetic nanoparticles as a heating source for magnetic particle hyperthermia it is of particular interest to know if the particles are free to move in the interstitial fluid or are fixed to the tumour tissue. The immobilization state determines the relaxation behaviour of the administered particles and thus their specific heating power. To investigate this behaviour, magnetic multicore nanoparticles were injected into experimentally grown tumours in mice and magnetic heating treatment was carried out in an alternating magnetic field (H = 25 kA m(-1), f = 400 kHz). The tested particles were well suited for magnetic heating treatment as they heated a tumour of about 100 mg by about 22 K within the first 60 s. Upon sacrifice, histological tumour examination showed that the particles form spots in the tissue with a mainly homogeneous particle distribution in these spots. The magnetic ex vivo characterization of the removed tumour tissue gave clear evidence for the immobilization of the particles in the tumour tissue because the particles in the tumour showed the same magnetic behaviour as immobilized particles. Therefore, the particles are not able to rotate and a temperature increase due to Brown relaxation can be neglected. To accurately estimate the heating potential of magnetic materials, the respective environments influencing the nanoparticle mobility status have to be taken into account.

An in vitro characterization study of new near infrared dyes for molecular imaging

The spectroscopic properties, stability, and cytotoxicity of series of cyanine labels, the dyes DY-681, DY-731, DY-751, and DY-776, were studied to identify new tools for in vivo fluorescence imaging and to find substitutes for DY-676 recently used by us as fluorescent label in a target-specific probe directed against carcinoembryonic antigen (CEA). This probe enables the selective monitoring of CEA-expressing tumor cells in mice, yet displays only a low fluorescence quantum yield and thus, a non-optimum sensitivity. All the DY dyes revealed enhanced fluorescence quantum yields, a superior stability, and a lower cytotoxicity in comparison to clinically approved indocyanine green (ICG). With DY-681 and far-red excitable DY-731 and DY-751, we identified three dyes with improved properties compared to DY-676 and ICG.

Suitable Labels for Molecular Imaging – Influence of Dye Structure and Hydrophilicity on the Spectroscopic Properties of IgG Conjugates

Aiming at the design of highly brilliant NIR emissive optical probes, e.g., for in vivo near-infrared fluorescence imaging (NIRF), we studied the absorption and fluorescence properties of the asymmetric cyanines Dy678, Dy681, Dy682, and Dy676 conjugated to the model antibody IgG. The ultimate goal was here to derive general structure-property relationships for suitable NIR fluorescent labels. These Dy dyes that spectrally match Cy5 and Cy5.5, respectively, were chosen to differ in chromophore structure, i.e., in the substitution pattern of the benzopyrylium end group and in the number of sulfonic acid groups. Spectroscopic studies of the free and IgG-bound fluorophores revealed a dependence of the obtained dye-to-protein ratios on dye hydrophilicity and control of the fluorescence quantum yields (Φ(f)) of the IgG conjugates by the interplay of different fluorescence reduction pathways like dye aggregation and fluorescence resonance energy transfer (FRET). Based upon aggregation studies with these dyes, the amount of dye dimers in the IgG conjugates was determined pointing to dye hydrophilicity as major parameter controlling aggregation. To gain further insight into the exact mechanism of dye dimerization at the protein, labeling experiments at different reaction conditions but constant dye-to-protein ratios in the reaction solution were performed. With Dy682 that displays a Φ(f) of 0.20 in PBS and 0.10 for moderate dye-to-protein ratio of 2.5, a low aggregation tendency, and a superior reactivity in IgG labeling, we identified a promising diagnostic tool for the design of NIR fluorescent probes and protein conjugates.

Efficient treatment of breast cancer xenografts with multifunctionalized iron oxide nanoparticles combining magnetic hyperthermia and anti-cancer drug delivery

IntroductionTumor cells can effectively be killed by heat, e.g. by using magnetic hyperthermia. The main challenge in the field, however, is the generation of therapeutic temperatures selectively in the whole tumor region. We aimed to improve magnetic hyperthermia of breast cancer by using innovative nanoparticles which display a high heating potential and are functionalized with a cell internalization and a chemotherapeutic agent to increase cell death.MethodsThe superparamagnetic iron oxide nanoparticles (MF66) were electrostatically functionalized with either Nucant multivalent pseudopeptide (N6L; MF66-N6L), doxorubicin (DOX; MF66-DOX) or both (MF66-N6LDOX). Their cytotoxic potential was assessed in a breast adenocarcinoma cell line MDA-MB-231. Therapeutic efficacy was analyzed on subcutaneous MDA-MB-231 tumor bearing female athymic nude mice.ResultsAll nanoparticle variants showed an excellent heating potential around 500 W/g Fe in the alternating magnetic field (AMF, conditions: H = 15.4 kA/m, f = 435 kHz). We could show a gradual inter- and intracellular release of the ligands, and nanoparticle uptake in cells was increased by the N6L functionalization. MF66-DOX and MF66-N6LDOX in combination with hyperthermia were more cytotoxic to breast cancer cells than the respective free ligands. We observed a substantial tumor growth inhibition (to 40% of the initial tumor volume, complete tumor regression in many cases) after intratumoral injection of the nanoparticles in vivo. The proliferative activity of the remaining tumor tissue was distinctly reduced.ConclusionThe therapeutic effects of breast cancer magnetic hyperthermia could be strongly enhanced by the combination of MF66 functionalized with N6L and DOX and magnetic hyperthermia. Our approach combines two ways of tumor cell killing (magnetic hyperthermia and chemotherapy) and represents a straightforward strategy for translation into the clinical practice when injecting nanoparticles intratumorally.

Diagnosis of Arthritis Using Near-infrared Fluorochrome Cy5.5

Purpose:Near-infrared range fluorescence (NIRF) imaging is a potential tool to diagnose biologic processes in vivo. This applicability study sought to define whether imaging with fluorochrome Cy5.5 can identify arthritis in murine antigen-induced arthritis (AIA). Materials and Methods:On day 7 of AIA (n = 9 mice), fluorescence intensities in inflamed and contralateral knee joints (the latter as internal control) were measured before and after intravenous injection of Cy5.5 (until 72 hours). Cy5.5 joint deposition was verified by confocal laser-scanning microscopy. Dye phagocytosis was evaluated in cultured macrophages (cell line PMJ2-R) by FACS analysis. Cy5.5 binding to serum protein was tested by NIRF scanning and gel electrophoresis. Results:Between 2 and 72 hours, the arthritic knee joints showed significantly higher fluorescence intensities compared with contralateral joints. Microscopy confirmed Cy5.5 deposition in the synovial membrane. Cultured macrophages actively phagocytosed Cy5.5. Cy5.5 bound mainly to albumin as the main serum protein. Conclusion:NIRF imaging with Cy5.5 can identify arthritic joints in vivo, likely due to nonspecific deposition.

In vivo applications of magnetic nanoparticle hyperthermia

Abstract Hyperthermia is considered to be a promising tool for the treatment of tumours. Intensive research activities reveal a distinct impact not only on the cellular level but also on tumour physiology which favours the combination with the classical oncologic modalities radio- and chemotherapy. Different techniques have been established so far. Among them, magnetic hyperthermia exploits the intrinsic magnetic properties of iron oxide nanoparticles (magnetite and maghemite) which induce heating during the exposure to an alternating magnetic field. Beyond the advantage that heating is generated within the tumour and not from outside the body, the amounts of magnetic material and their intratumoral distribution patterns are key factors determining the therapeutic outcome. They can be influenced by the use of different application routes, which will be discussed in this paper.