Functional magnetic resonance (fMR) imaging of a rat brain tumor model: implications for evaluation of tumor microvasculature and therapeutic response.

Functional MR (fMR) imaging techniques based on blood oxygenation level dependent (BOLD) effects were developed and applied to a rat brain tumor model to evaluate the potential utility of the method for characterizing tumor growth and regression following treatment. Rats bearing 9L brain tumors in situ were imaged during inhalation of room air and after administration of 100% oxygen + acetazolamide (ACZ) injected 15 mg/kg intravenously. Pixel-to-pixel fMR maps of normalized signal intensity change from baseline values were calculated from T2 weighted spin echo (SE) images acquired pre- and post- oxygen + ACZ administration. Resultant fMR maps were then compared to gross histological sections obtained from corresponding anatomical regions. Regions containing viable tumor with increased cellular density and localized foci of necrotic tumor cells consistent with hypoxia were visualized in the fMR images as regions with decreased signal intensities, indicating diminished oxyhemoglobin concentration and blood flow as compared to normal brain. Histological regions having peritumor edema, caused by increased permeability of tumor vasculature, were visualized in the fMR images as areas with markedly increased signal intensities. These results suggest that fMR imaging techniques could be further developed for use as a non-invasive tool to assess changes in tumor oxygenation/hemodynamics, and to evaluate the pharmacologic effect of anti-neoplastic drugs.

Liposome-mediated therapy of intracranial brain tumors in a rat model.

PURPOSE: Malignant brain tumors represent a serious therapeutic challenge, and survival often is low. We investigated the delivery of doxorubicin (DXR) to rat brain tumors in situ via liposomes, to test the hypothesis that intact liposomes undergo deposition in intracranial tumor through a compromised blood-tumor vasculature. Both therapeutic effect and intra-tumor drug carrier distribution were evaluated to identify variables in carrier-mediated delivery having impact on therapy. METHODS: The rat 9L gliosarcoma tumor was implanted orthotopically in Fischer 344 rats in the caudate-putamen region. The tumor-bearing rats were treated with DXR, either free or encapsulated in long-circulating, sterically-stabilized liposomes. Anti-tumor efficacy was assessed by survival time. In parallel, liposomes labeled with a fluorescent phospholipid analog were injected into tumor-bearing rats. At predetermined intervals, the brains were perfused with fixative, sectioned, and imaged with laser scanning confocal microscope (LSCM) to investigate the integrity of the tumor vascular bed and the intratumor deposition of liposomes. RESULTS: Free DXR given in 3 weekly iv injections was ineffective in increasing the life span of tumor-bearing rats at cumulative doses < or = 17 mg/kg, and at the highest dose (17 mg/kg) decreased survival slightly, compared to saline-treated controls. In contrast, DXR encapsulated in long-circulating liposomes mediated significant increases in life span at 17 mg/kg. Rats showed a 29% percent increase in median survival, respectively, compared to saline-control animals. The delay of treatment after tumor implantation was a major determinant of therapeutic effect. Fluorescent liposomes were deposited preferentially in tumor rather than normal brain, and were distributed non-uniformly, in close proximity to tumor blood vessels. CONCLUSIONS: Liposomes can be used to enhance delivery of drugs to brain tumors and increase therapeutic effect. The therapeutic effect may arise from release of drug from liposomes extravasated in discrete regions of the tumor vasculature and the extravascular space.

Boronated metalloporphyrins: a novel approach to the diagnosis and treatment of cancer using contrast-enhanced MR imaging and neutron capture therapy.

Porphyrins are a unique class of metal chelating agents that have shown specific affinity for neoplasms. The water-soluble free-base derivative, tetrakiscarborane carboxylate ester of 2,4-(alpha,beta-dihydroxyethyl) deuteroporphyrin IX (BOPP), an agent designed for neutron capture therapy, has previously demonstrated selective localization and retention in a C6 murine glioma. In the present work, the authors demonstrate that the manganese chelate of BOPP also selectively localizes in a rat 9L gliosarcoma and preferentially enhances the tumor-normal brain contrast of T1-weighted images for at least 92 hours. The data indicate a maximal enhancement of contrast between tumor and normal brain at 24 hours after injection, compared with 5 minutes for manganese (III) tetraphenylporphine sulfonate (TPPS4). The results also indicate that Mn-BOPP may have a slower uptake in the 9L glioma than Mn-TPPS4 but a longer retention in the tumor. Mn-BOPP is unique in that it represents, to the authors' knowledge, the first example of a single agent that can enhance contrast between tumor and normal tissue and be potentially effective as an agent for boron neutron capture therapy.

A microcomputer based system for three-dimensional reconstructions from tomographic or histologic sections.

The reconstructions of three-dimensional (3-D) objects from serial two-dimensional (2-D) images can contribute to the understanding of many biologic structures, from organelles to organs and tissues. The 3-D reconstruction of sections can be divided into several major tasks: image acquisition, alignment of slices, internal object definition, object reconstruction and rotation of the completed image. A fast, versatile, interactive system was devised for the reconstruction of 3-D objects from serial 2-D images using a low-cost microcomputer, original programs and commercial software. The system allows reconstruction from any serial images, e.g., electron micrographs, histologic sections or computed tomograms. A photographic image or a microscopic field is acquired into the computer memory using a video digitizer. Slices are superimposed and aligned to each other using an operator-interactive program. A contour-(edge-) finding algorithm isolates an object of interest from the background image by "subtraction" of the image from an overlaid, slightly shifted identical image. Contours for each slice are input to a reconstruction procedure, which calculates the x, y and z coordinates of every point in a slice and the thickness and number of slices. It then calculates the illumination for every point using a given point source of light and an intensity-fading coefficient. Finally, the points are represented by cubes to provide dimension and reflective surfaces. A cube of appropriate shade and color represents in 2-D the equivalent of a 3-D object; this results in a very effective 3-D image. The reconstruction is rotated by recalculating the positions of every point defining the object and rebuilding the image.(ABSTRACT TRUNCATED AT 250 WORDS)

A microcomputer-based algorithm for digitized image simplification and information extraction.

Spatial analysis of objects often requires significant image simplification prior to information extraction and application of a decision-making algorithm. Much decision making based on images (e.g., histologic diagnoses) requires identifying patterns in complex backgrounds (image simplification) and comparison of those patterns to other patterns (decision making). Automated extraction of information from images commonly requires the extraction system to recognize edges (contours) of structures and their internal discontinuities (such as gradations in density) and to selectively suppress irrelevant data in order to conserve memory and speed computation; data from homogeneous image areas occupy memory, but are noncontributory or redundant. This paper describes the development of a microcomputer-based algorithm that deletes all homogeneous information from overlaid digitized images, generating contours in the place of nonhomogeneities. Contours corresponding to different areas or objects depend on color differences between an object and its surroundings. Any set of contours can be deleted almost instantaneously, leaving only those of interest. Contours can be highlighted by an operator-driven interactive process if desired and can be deleted and retrieved until an appropriate image is obtained. This contour-generating and image-simplification algorithm facilitates three-dimensional reconstruction of an object from serial images by reducing the number of calculations required and yielding a cleaner final image.