Search

The development of efficient and biocompatible contrast agents is particularly urgent for modern clinical surgery. Nanostructured materials raised great interest as contrast agents for different imaging techniques, for which essential features are high contrasts, and in the case of precise clinical surgery, minimization of the signal spatial dispersion when embedded in biological tissues. This study deals with the development of a multimodal contrast agent based on an injectable hydrogel nanocomposite containing a lanthanide-activated layered double hydroxide coupled to a biocompatible dye (indocyanine green), emitting in the first biological window. This novel nanostructured thermogelling hydrogel behaves as an efficient tissue marker for optical and magnetic resonance imaging because the particular formulation strongly limits its spatial diffusion in biological tissue by exploiting a simple injection. The synergistic combination of these properties permits to employ the hydrogel ink simultaneously for both optical and magnetic resonance imaging, easy monitoring of the biological target, and, at the same time, increasing the spatial resolution during a clinical surgery. The biocompatibility and excellent performance as contrast agents are very promising for possible use in image-guided surgery, which is currently one of the most challenging topics in clinical research.

The article concerns the obtainment of liposomal doxorubicin (Dox) in which liposomes are externally modified with a targeting peptide able to drive the formulation in a selective way on membrane receptors overexpressed in tumors. We developed a kit composed by three different vials: (A) a vial containing a sterile, translucent, red dispersion of the liposomal doxorubicin drug (Doxil®), (B) a vial filled with a lyophilized powder of a modified phospholipid with a reactive function (DSPE-Peg-maleimide), and (C) a vial containing a 1–9 bombesin peptide analogue (Cys-BN-AA1) chemically modified to react in stoichiometric ratio respect to DSPE-Peg-maleimide. The chosen peptide is a stable analogue antagonist of the wild-type 1–9 bombesin peptide; it is very stable in serum; maintains high specificity, with nanomolar affinity, towards gastrin release peptide receptors (GRPRs indicated also as BB2); and is overexpressed in some cancer cells. Results on animal studies clearly indicate that in mice treated with the kit product (i.e., pegylated liposomal Dox modified with the bombesin analogue, Doxil-BN-AA1), tumor growth is reduced, with an improved efficacy respect to mice treated with non-modified pegylated liposomal Dox or with saline solution.

AbstractObjective: We studied ob/ob and wild-type (WT) mice to characterize the adipose tissues depots and other visceral organs and to establish an experimental paradigm for in vivo phenotyping.Research Methods and Procedures: An in vivo evaluation was conducted using magnetic resonance imaging and 1H-magnetic resonance spectroscopy (1H-MRS). We used T1-weighted images and three-dimensional spin echo T1-weighted images for the morphological analysis and 1H-MRS spectra on all body mass, as well as 1H-MRS spectra focalized on specific lipid depots [triglyceride (TG) depots] for a molecular analysis.Results: In ob/ob mice, three-dimensional evaluation of the trunk revealed that ∼64% of the volume consists of white adipose tissue, which is 72% subcutaneous and 28% visceral. In vivo 1H-MRS showed that 20.00 ± 6.92% in the WT group and 58.67 ± 6.65% in the ob/ob group of the total proton content is composed of TG protons. In in vivo-localized spectra of ob/ob mice, we found a polyunsaturation degree of 0.5247 in subcutaneous depots. In the liver, we observed that 48.7% of the proton signal is due to water, whereas in the WT group, the water signal amounted to 82.8% of the total proton signal. With the sequences used, the TG amount was not detectable in the brain or kidneys.Discussion: The present study shows that several parameters can be obtained by in vivo examination of ob/ob mice by magnetic resonance imaging and 1H-MRS and that the accumulated white adipose tissue displays low polyunsaturation degree and low hydrolipidic ratio. Relevant anatomical alterations observed in urinary and digestive apparatuses should be considered when ob/ob mice are used in experimental paradigms.

The aim of this study was to compare the efficacy of gadoteridol, B22956/1 (a new protein binding blood pool contrast agent), and (Gd-DTPA)37-albumin in detecting, by dynamic contrast-enhanced magnetic resonance imaging (MRI), the effect in vivo of tamoxifen in an experimental model of breast tumor implanted in rats.

This study was designed to evaluate the influence of age on the dynamic contrast-enhanced MRI of ischemic tissue. A well-established model of peripheral arterial insufficiency (i.e., the rat hindlimb ischemia after removal of femoral artery) in different age groups (i.e., young, presenescent, and senescent rats) was studied. The analysis of the MR signal demonstrated a marked accumulation of a contrast agent (Gd-DTPA) in the ischemic leg (ischemia-related enhancement, IRE). IRE was an age-related event: 4-month-old rats showed a strong IRE while 12-month-old rats and 20-month old rats showed a significantly reduced IRE in comparison to young animals. Histological analysis of the ischemic muscles revealed that there was no evidence of significant necrosis of the muscle tissue but only a weak interstitial fibrosis; CD31-immunostaining revealed a preserved microcirculatory bed. Magn Reson Med 49:386–390, 2003. © 2003 Wiley-Liss, Inc.

This study was designed to evaluate the influence of age on the dynamic contrast‐enhanced MRI of ischemic tissue. A well‐established model of peripheral arterial insufficiency (i.e., the rat hindlimb ischemia after removal of femoral artery) in different age groups (i.e., young, presenescent, and senescent rats) was studied. The analysis of the MR signal demonstrated a marked accumulation of a contrast agent (Gd‐DTPA) in the ischemic leg (ischemia‐related enhancement, IRE). IRE was an age‐related event: 4‐month‐old rats showed a strong IRE while 12‐month‐old rats and 20‐month old rats showed a significantly reduced IRE in comparison to young animals. Histological analysis of the ischemic muscles revealed that there was no evidence of significant necrosis of the muscle tissue but only a weak interstitial fibrosis; CD31‐immunostaining revealed a preserved microcirculatory bed. Magn Reson Med 49:386–390, 2003. © 2003 Wiley‐Liss, Inc.

The intra‐orbital lachrymal gland (Harderian gland, or HG) of the female rat was studied by magnetic resonance imaging (MRI) to evaluate whether MRI can be used to visualize the gland in vivoand localized‐1H‐spectroscopy detect its lipid content. The results were correlated with post‐mortem anatomical sections, and with light and electron microscopy. On MRI, HG presented as a mass located between the ocular bulb and the orbit. In strongly T2W sequences the secretory structures had a reduced signal while intraparenchymal connective tissue was visible. T2‐quantitative maps values of HG (60.12 ± 8.15 ms, mean ± SD) were different from other tissues (i.e. muscular tissue, T2 = 44.79 ± 3.43 ms and olfactory bulb, T2 = 79.26 ± 4.25 ms). In contrast‐enhanced‐MRI, HG had a signal‐intensity‐drop of 0.074 ± 0.072 (mean ± SD), after injection of AMI‐25, significantly different from the muscle (0.17 ± 0.10). Localized MRI spectra gave a large part of the signal originating from fat protons, but with a significant percentage from water protons. At light and electron microscopy the lipid deposition appeared to be composed of low‐density material filling a large part of the cytoplasm, and the porphyrin aggregates were easily recognizable. The data demonstrate that an in vivostudy of the HG was feasible and that high‐field MRI allowed analysis of the gross anatomy detecting the lipid content of the gland.

Modifications in the contralateral “healthy” hemisphere in a population of rats bearing cortical infarction were studied in vivo by magnetic resonance imaging (MRI) with the aim to investigate whether cerebral areas not directly involved in the lesion react at the presence of an ischemic lesion. The study was performed in rats in which a transtemporal approach was adopted to occlude the right middle cerebral artery (MCA). For MRI, the animals were examined at 4.7 Tesla and quantitative T2 parametric images were obtained by a multiecho sequence. Healthy rats and sham-operated animals were used as control groups. The quantitative T2 parametric images showed that in the first week after the ischemia a significant increase in the mean T2 was seen in the lesioned parietal cortex, compared to the corresponding region of healthy rats (106 msec vs. 68 msec, P < 0.001). The contralateral “healthy” hemisphere showed T2 mean values not significantly different from the corresponding hemisphere of healthy rats (71 msec vs. 70 msec). However, a statistically significant increase in the T2 values was evident in the hypothalamic region (74 msec vs. 66 msec, P < 0.001). In rats examined 1 month after the ischemia, the T2 values of the hypothalamus were lower than those observed one week after ischemia (69 msec) but remained higher than in controls. The present study demonstrates that after a cerebral ischemia areas of secondary involvement distant from the lesion are present and can be studied in vivo by quantitative MRI. Anat Rec 266:118–122, 2002. © 2002 Wiley-Liss, Inc.

Modifications in the contralateral “healthy” hemisphere in a population of rats bearing cortical infarction were studied in vivo by magnetic resonance imaging (MRI) with the aim to investigate whether cerebral areas not directly involved in the lesion react at the presence of an ischemic lesion. The study was performed in rats in which a transtemporal approach was adopted to occlude the right middle cerebral artery (MCA). For MRI, the animals were examined at 4.7 Tesla and quantitative T2 parametric images were obtained by a multiecho sequence. Healthy rats and sham‐operated animals were used as control groups. The quantitative T2 parametric images showed that in the first week after the ischemia a significant increase in the mean T2 was seen in the lesioned parietal cortex, compared to the corresponding region of healthy rats (106 msec vs. 68 msec, P< 0.001). The contralateral “healthy” hemisphere showed T2 mean values not significantly different from the corresponding hemisphere of healthy rats (71 msec vs. 70 msec). However, a statistically significant increase in the T2 values was evident in the hypothalamic region (74 msec vs. 66 msec, P< 0.001). In rats examined 1 month after the ischemia, the T2 values of the hypothalamus were lower than those observed one week after ischemia (69 msec) but remained higher than in controls. The present study demonstrates that after a cerebral ischemia areas of secondary involvement distant from the lesion are present and can be studied in vivo by quantitative MRI. Anat Rec 266:118–122, 2002. © 2002 Wiley‐Liss, Inc.

Parametric mapping of polyunsaturated fatty acids (PUFA) distribution in adipose tissues was obtained by ¹H chemical shift imaging (CSI). A matrix of spectra, acquired with a CSI sequence having two spatial and one spectroscopic dimension, was processed with ad hoc algorithms. The protocol was applied to phantoms containing different lipids in which the degree of polyunsaturation was determined by high-resolution nuclear magnetic resonance (NMR). High correlation (R² = 0.998) between degrees of polyunsaturation given by our protocol and that measured by high-resolution NMR was found. The thoracic region of rats was also examined. Parametric maps of the polyunsaturation degree were obtained for the brown adipose tissue and the white axillary fat: the first deposit was found more polyunsaturated than the second. Finally, in vivo mapping of the inguinal region of the rat was produced that allowed us to individuate PUFA-rich areas in adipose tissue. This work demonstrates the feasibility of PUFA imaging in vivo. Magn Reson Med 46:879–883, 2001. © 2001 Wiley-Liss, Inc.

Parametric mapping of polyunsaturated fatty acids (PUFA) distribution in adipose tissues was obtained by 1H chemical shift imaging (CSI). A matrix of spectra, acquired with a CSI sequence having two spatial and one spectroscopic dimension, was processed with ad hoc algorithms. The protocol was applied to phantoms containing different lipids in which the degree of polyunsaturation was determined by high‐resolution nuclear magnetic resonance (NMR). High correlation (R2= 0.998) between degrees of polyunsaturation given by our protocol and that measured by high‐resolution NMR was found. The thoracic region of rats was also examined. Parametric maps of the polyunsaturation degree were obtained for the brown adipose tissue and the white axillary fat: the first deposit was found more polyunsaturated than the second. Finally, in vivo mapping of the inguinal region of the rat was produced that allowed us to individuate PUFA‐rich areas in adipose tissue. This work demonstrates the feasibility of PUFA imaging in vivo. Magn Reson Med 46:879–883, 2001. © 2001 Wiley‐Liss, Inc.

The possible persistence of a microvascular deficit at long time intervals after cerebral ischemia induction is not well established. In rats, we have generated in vivomaps of the regional cerebral blood volume (rCBV) at different time intervals after middle cerebral artery occlusion (MCAo) with the aim to evaluate the persistence of a rCBV deficit in the damaged area or in the surrounding regions. The rats were examined by magnetic resonance imaging (MRI) at different time intervals, starting from the first day until three months after ischemia and postmortemhistological and ultrastructural correlation was obtained. All MRI experiments were carried out using an imager-spectrometer equipped with a 4.7 Tesla magnet. To produce the susceptibility-weighted rCBV images, a suspension of superparamagnetic iron oxide nanoparticles (AMI-25) was injected to the rat. In a control group (nonoperated or sham-operated rats), a symmetrical distribution of rCBV values was found between the two hemispheres (differences between left and right cortex below 8%). In the rats with MCAo an evident vascular asymmetry was found 24 h after ischemia (differences between left and right ranging from 22 and 77%) and reduced rCBV values were evident in the ischemic areas. In a time range following the 15th day most of the rats showed a complete recovery of the lesion while only four animals still had a small residual lesion, as probed by T2-weighted (T2W) images. In three of these four cases, the reduction of rCBV in the ipsilateral cortex with respect to the contralateral was greater than 20%. Correlation was found (Y> 0.8) between late rCBV measurement and the initial volume of the lesion (hyperintense region in T2W images). The postmortem measurements correlate much better with the rCBV data than with the T2W ones. In conclusion, the present work demonstrates that cortical lesions may result in a deficit of rCBV for long periods and that a mismatch between T2w and rCBV data can be present during the repair process.

We propose an experimental protocol, based on chemical shift magnetic resonance imaging (CSI) that improves the methods presently available for the in vivo study of the thymus in small animals. Male Balb/c mice were examined in an imager-spectrometer equipped with a 4.7 T magnet. Three groups of animals with different ages were used: the first group consisted of 3-month-old mice (n = 5), the second group of 19-month-old mice (n = 5), and the third group of 26-month-old mice (n = 4). The identification of thymic parenchyma was obtained by two (T1-weighted spin-echo and CSI water-selective) images. The T1-weighted spin-echo image provided a detailed anatomical description of the organs located in the thorax. The CSI water-selective image provided a detailed description of thymic location, shape, and dimensions. The cross-sectional area of the thymus, measured from CSI images, showed a decreasing trend with increasing age. The values of the thymus-muscle contrast-to-noise ratio were measured in both spin echo and CSI images. While the contrast between thymus and muscle was greatly improved in the young and presenescent group, the difference was not statistically significant in the senescent group. In conclusion, the proposed method allows the study of thymic modification during the passage from young to pre-senescent age and from pre-senescent to old age. This method could be useful in studies in which experimental manipulation or drug treatments produce changes in the dimension and fat content of this organ. The proposed protocol, based on CSI, appears to be an improved methodology for study of the thymus.J. Magn. Reson. Imaging 1999;10:97–101. © 1999 Wiley-Liss, Inc.

We propose an experimental protocol, based on chemical shift magnetic resonance imaging (CSI) that improves the methods presently available for the in vivo study of the thymus in small animals. Male Balb/c mice were examined in an imager‐spectrometer equipped with a 4.7 T magnet. Three groups of animals with different ages were used: the first group consisted of 3‐month‐old mice (n= 5), the second group of 19‐month‐old mice (n= 5), and the third group of 26‐month‐old mice (n= 4). The identification of thymic parenchyma was obtained by two (T1‐weighted spin‐echo and CSI water‐selective) images. The T1‐weighted spin‐echo image provided a detailed anatomical description of the organs located in the thorax. The CSI water‐selective image provided a detailed description of thymic location, shape, and dimensions. The cross‐sectional area of the thymus, measured from CSI images, showed a decreasing trend with increasing age. The values of the thymus‐muscle contrast‐to‐noise ratio were measured in both spin echo and CSI images. While the contrast between thymus and muscle was greatly improved in the young and presenescent group, the difference was not statistically significant in the senescent group. In conclusion, the proposed method allows the study of thymic modification during the passage from young to pre‐senescent age and from pre‐senescent to old age. This method could be useful in studies in which experimental manipulation or drug treatments produce changes in the dimension and fat content of this organ. The proposed protocol, based on CSI, appears to be an improved methodology for study of the thymus.J. Magn. Reson. Imaging 1999;10:97–101. © 1999 Wiley‐Liss, Inc.