Search

The implantation of breast prostheses for both aesthetic and reconstructive purposes has been growing exponentially in the last 20 years. Safety and prosthesis lifespan are majorly debated issues in relation to the correlated long-term complications. Mainly the periprosthetic capsule that develops around the implant is often the cause of complications and particularly for macrotextured silicone breast implants. Some reports have tried to elucidate the mechanism by which macrotextured silicone implants undergo damage and cause double capsule formation. In this study, we investigated the morphological characteristics of double capsule of macrotextured implants surgically removed from patients. With the use of microscopy techniques, this work analyzed the newly formed tissue observed in the interaction between synthetic and biological surfaces.

About twenty years ago, nanotechnology began to be applied to biomedical issues giving rise to the research field called nanomedicine. Thus, the study of the interactions between nanomaterials and the biological environment became of primary importance in order to design safe and effective nanoconstructs suitable for diagnostic and/or therapeutic purposes. Consequently, imaging techniques have increasingly been used in the production, characterisation and preclinical/clinical application of nanomedical tools. This work aims at making an overview of the microscopy and imaging techniques in vivo and in vitro in their application to nanomedical investigation, and to stress their contribution to this developing research field.

Background Tissue expanders are widely utilized in plastic surgery. Traditional expanders usually are “inflatable balloons,” which are planned to grow additional skin and/or to create space to be filled, for example, with an implant. In very recent years, reports suggest that negative pressure created by an external device (ie, Brava) induces both skin expansion and adipogenesis. Objectives The authors evaluated and assessed the adipogenetic potential of a novel internal tissue expander in an in vivo animal model. Methods New Zealand female rabbits were enrolled in the study. A prototype spiral inner tissue expander was employed. It consisted of a-dynamic conic expander (DCE) with a valve at the end: when empty, it is flat (Archimedean spiral), whereas when filled with a fluid, it takes a conic shape. Inside the conic spiral, a negative pressure is therefore created. DCE is implanted flat under the latissimus dorsi muscle in experimental animals (rabbit) and then filled to reach the conical shape. Animals were investigated with magnetic resonance imaging, histology, and transmission electronic microscopy at 3, 6, and 12 months. Results Magnetic resonance imaging revealed a marked increase in newly formed adipose tissue, reaching its highest amount at 12 months after the DCE implantation. Histology confirmed the existence of new adipocytes, whereas transmission electronic microscopy ultrastructure confirmed that most of these new cells were mature adipocytes. Conclusions Tensile stress, associated with negative-pressure expanders, generated newly white subcutaneous adipose tissue.

Interstitial lung involvement in Systemic Sclerosis (SSc-ILD) is a complication with high morbidity and mortality. Specifically, engineered gold nanoparticles (GNPs) are proposed as targeted delivery system increasing efficacy of drugs with antifibrotic effect, such as tyrosine kinases. We aimed to test in vitro and in vivo the activity of targeted Imatinib (Im)-loaded GNP on SSc-ILD patients derived cells and in experimental model of lung fibrosis. GNPs functionalized with anti-CD44 and loaded with Im (GNP-HCIm) were synthesized. Lung fibroblasts (LFs) and alveolar macrophages from bronchoalveolar lavage fluids of SSc-ILD patients were cultured in presence of nanoparticles. GNP-HCIm significantly inhibited proliferation and viability inducing apoptosis of LFs and effectively reduced IL-8 release, viability and M2 polarization in alveolar macrophages. Anti-fibrotic effect of tracheal instilled GNP-HCIm was evaluated on bleomycin lung fibrosis mouse model comparing effect with common route of Im administration. GNP-HCIm were able to reduce significantly lung fibrotic changes and collagen deposition. Finally, electron microscopy revealed the presence of GNPs inside alveolar macrophages. These data support the use of GNPs locally administered in the development of new therapeutic approaches to SSc-ILD.

Object Pancreatic cancer is one of the most lethal human cancer and appropriate experimental tumor models are needed for the development of innovative therapeutic approaches. This paper describes an experimental model of human pancreatic cancer and a related non invasive imaging technique suitable for monitoring tumor growth and metastatization. The aim of the work was the implementation of an experimental platform suitable for assessing the efficacy of new therapeutic agents. Materials and methods Human pancreatic cancer cells (PANC-1-Luc +) were injected into the pancreas of female athymic CD1 mice. Magnetic Resonance Imaging (MRI) at 4.7 T and Bioluminescence Imaging (BLI) were performed in each mouse at three time points after cell inoculation (1, 2 and 3 months). Two groups of mice were studied: the first group of n = 13 mice in which 5 * 10 6 cells were injected and the second group of n = 10 mice in which 2 * 10 6 cells were injected. MRI examination included T2w acquisitions and (at the last time point) Dynamic-contrast-enhanced-MRI (DCE-MRI). Results Each mouse underwent three longitudinal MRI and BLI examinations. BLI was more sensitive than MRI producing higher detection rate at early time points. Moreover in one case of abdominal dissemination of pancreatic tumor cells, small tumoral masses were detected by BLI and not detected by MRI. However BLI appears more prone to experimental error most likely due to photon attenuation. In 4 mice BLI produced false negative results. DCE-MRI experiments providing information on tumor perfusion were conducted successfully in this anatomical district and demonstrated that the tumor tissues from the second experimental group are more vascularized compared to the first group. Conclusion The present study performed on the experimental model of pancreatic cancer here described shows that MRI and BLI are complementary techniques and that synergistic application of both can overcome the intrinsic limitations of each.

Background Systemic Sclerosis (SSc) is an autoimmune disorder frequently affected by an interstitial lung involvement (ILD) that significantly deteriorates long term outcomes. In previous experiments we proved that specifically engineered gold-nanoparticles (GNP) loaded with imatinib and targeted with an anti CD44 Ab (GNP-HCim) significantly inhibited proliferation and induced apoptosis of fibroblast-like cells derived from ILD-SSc patients [1]. In vitro, GNP-HCim showed higher efficacy compared to the drug alone. Objectives To demonstrate in vivo the efficacy of GNP-HCim in ameliorating bleomycin-induced lung fibrosis Methods Eight-week-old C57BL6 male mice (n=8/group) were assigned to either: (1) controls receiving intratracheal aereosolization of saline solution and unloaded functionalised GNP (GNP-HC); (2) mice treated with intratracheal instillation of bleomycin (50 uL) on day 0 and GNP-HC; (3) mice treated with bleomycin on day 0 plus GNP-HCim; (4) mice treated with bleomycin plus intraperitoneal (i.p.) Imatinib (50 mg/kg, once daily). GNP-HC or GNP-HCim were administered by intratracheal instillation on day 10–15–20–25 and 3 h before culling. All mice were sacrificed on day 28. Lung specimens were analysed by electron microscopy, immunohistochemistry and immunofluorescence (IF). Data were evaluated by 2 blind observers and analysed with GraphPrism software for statistics. Results The administration of imatinib i.p or via GNP-HC reduced pathologic changes of the lungs as evaluated by the Lung Injury score and the Ashcroft score (p Conclusions In the experimental model of bleomycin-induced lung fibrosis imatinib delivered to lungs through inhalation of anti CD44 targeted GNP was as effective as imatinib administered by i.p. route. These data favour the use of GNP in the development of new therapeutic approaches to SSc-ILD, which might be associated to a lower toxicity and side effects of systemic treatment. References Cova E, Ann rheum Dis 2016, 75(Suppl2): 61. Disclosure of Interest None declared

Structural and functional MRI was used in conjunction with computerized electron microscopy morphometry to study changes 2 h, 24 h and 3 days after 4-aminopyridine-induced seizures lasting 2 h in rats. T2 (relaxation time) values showed changes throughout the cerebral cortex, hippocampus, amygdala and medial thalamus, with a different temporal progression, showing a complete recovery only after 3 days. Two hours after seizures, the apparent diffusion coefficient was decreased throughout the brain compared to control animals, and a further decrease was evident 24 h after seizures. This was followed by a complete recovery at 3 days post-seizures. Functional MRI was performed using regional cerebral blood volume (rCBV) maps. The rCBV was increased shortly after convulsions (2 h) in all structures investigated, with a significant return to baseline values in the parietal cortex and hippocampus, but not in the medial thalamic nuclei, 24 h after seizure onset. No rCBV alterations were detected 3 days after seizures. Electron microscopy of tissue samples of parietal neocortex and hippocampus revealed prominent astrocytic swelling 2 h post-convulsions which decreased thereafter gradually. In conclusion, this experiment reports for the first time structural and functional brain alterations, lasting several hours, in 4-aminopyridine-treated rats after seizure onset. MRI approach combined with histological and ultrastructural analysis provided a clarification of the mechanisms involved in the brain acute response to ictal activity.

Objectives: 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) 3 7 -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. Materials and Methods: Tumors were induced by subcutaneous injection of 10 6 mammary adenocarcinoma cells (13762 MAT B III). Treatment with tamoxifen (or vehicle) started on day 4 after implantation. On day 10 after implantation, animals were observed by MRI using B22956/1 (or gadoteridol) and, 24 hours later, using (Gd-DTPA)37-albumin. Results: Dynamic contrast-enhanced magnetic resonance imaging data showed that tamoxifen treatment decreased vascular permeability to B22956/1, whereas no difference was detectable in permeability to gadoteridol or to (Gd-DTPA) 3 7 -albumin. No effect on fractional plasma volume was detected with either of contrast agents. Conclusions: B22956/1 is superior to both small Gd chelates and macromolecular contrast agents in the assessment of the effect of tamoxifen treatment on tumor vasculature.

Purpose To compare T1- or T2-weighted magnetic resonance imaging (MRI) and ultrasonography (US) as tools for in vivo mapping of different tissue components in spontaneous tumors of transgenic mice. Materials and Methods Human-like mammary adenocarcinomas from FVB/neuT transgenic mice were analyzed by T2-weighted and T1-weighted MRI at 4.7 Tesla and US and then, after excision, were paraffin-embedded for histologic analysis. The histologic samples were prepared taking care to obtain sections that spatially matched the MRI and US images as precisely as possible. Results US can obtain basic information such as the size of developing tumors in experimental animals and can identify necrotic areas. T2-weighted MRI, especially if compared to T1-weighted MRI and/or US, allows advanced analysis of morphologic aspects, with high resolution in the differentiation of details of necrotic areas such as coagulation, liquefaction, biphasic splitting of cysts, and fibrotic and lipidic infiltration. Conclusion Of the three methods, T2-weighted MRI provides the most information about the anatomy of tumors. However, when distinctions between the different types of necrosis are not needed, US analysis is to be preferred for its practicality. J. Magn. Reson. Imaging 2004;19:570–579. © 2004 Wiley-Liss, Inc.

Spontaneous light emission from living animals can overcome the investigated light signals in small animal luminescence imaging. Despite autofluorescence emission is well studied the spontaneous luminescence background is less known and its importance is growing due to the new born imaging techniques like Cerenkov Luminescence Imaging and Radionuclide Luminescence Imaging in which faint sources are often involved. In order to investigate the spontaneous emission we studied the background luminescence in vivo from health Nu/Nu mice in optical imaging acquisitions and we related it with the optical properties of the diet of the animals. In particular luminescence images of mice feed with normal diet used in animal facilities were acquired using a commercial optical imager. The intensity and the spectral features of the luminescence emission from the animal surface after sunshine exposition and after normal lighting laboratory conditions were measured. The same was done with the pellets of food used to feed the animals. We found a background emission from the entire animal surface and localized light sources in the abdominal/lumbar region. Their intensity can be modulated by the light exposition of the animals before the imaging session and decreases along the time when they are put in darkness. The comparison of the luminescence time decay of animals and pellets suggests that the light sources are related to the persistent luminescence of the molecules contained in the food. So ambient exposure before imaging is important for luminescence imaging in order to keep down the background. The optical properties of food are also important and it necessary to check them before to feed the animals not only in fluorescence imaging but also in luminescence imaging.

Object The work is aimed to develop a murine model of rectal cancer, which could be used to monitor lymph node metastasis development by magnetic resonance imaging (MRI) and optical imaging (OI) techniques. Subjects and methods Ht-29 cancer cells were directly injected into the submucosal layer of the rectum of athymic nude mice using trans-anal rectal cancer cell injection (TARCI). Thirty-six mice were inoculated with 10 × 105 cells and five mice were treated with sterile phosphate buffer solution. One to 4 weeks after cell injection, tumor growth was evaluated in vivo using T2-weighted MRI at 4.7T. A further group of animal (n = 6) treated with ht-29_luc cells, with the same protocol, was monitored by optical imaging. In both groups, the presence of the primary tumor and of lymph nodes metastasis was confirmed by histology. Results In all animals, primary tumors were detectable by MRI, 1 week from TARCI. After 4 weeks primary tumors showed a mean longitudinal diameter of about 2 cm. All animals developed regional lymph node metastases. Others organs (e.g. lung or liver) were not affected. In fat-suppressed, T2-weighted MRI, lymph nodes appeared as small areas characterized by hyper-intense signal compared to muscle. OI permitted evaluation of the primary tumor growth in perineal region. Conclusions TARCI of ht-29 cells into the rectum of nude mice is a feasible way to obtain a easily reproducible model of regional lymph node metastases could be monitored by magnetic resonance and optical imaging techniques.

The main purpose of this work was to investigate both in vitro and in vivo Cherenkov radiation (CR) emission coming from 18F and 32P. The main difference between 18F and 32P is mainly the number of the emitted light photons, more precisely the same activity of 32P emits more CR photons with respect to 18F. In vitro results obtained by comparing beta counter measurements with photons average radiance showed that Cherenkov luminescence imaging (CLI) allows quantitative tracer activity measurements. In order to investigate in vivo the CLI approach, we studied an experimental xenograft tumor model of mammary carcinoma (BB1 tumor cells). Cherenkov in vivo dynamic whole body images of tumor bearing mice were acquired and the tumor tissue time activity curves reflected the well-known physiological accumulation of 18F-FDG in malignant tissues with respect to normal tissues. The results presented here show that it is possible to use conventional optical imaging devices for in vitro or in vivo study of beta emitters.

In this reply, we address the criticisms by Drs Mitchell, Gill and Cherry (Mitchell et al 2010 Phys. Med. Biol. 55 L43–4) regarding our paper recently published in Physics Medicine and Biology (Spinelli et al 2010 Phys. Med. Biol. 55 483–95). In our paper we showed that it is possible to acquire Cerenkov radiation images of mice injected with 18F-FDG using an optical imaging device. We also showed two different approaches that can be used in order to estimate the depth of the Cerenkov radiation source inside the mouse. More precisely, we will discuss the criticisms regarding the proposed model of the Cerenkov radiation spectrum for positron emitters and the estimation methods of the radiation source depth by providing further experimental results.

In this paper, we showed that Cerenkov radiation (CR) escaping from the surface of small living animals injected with (18)F-FDG can be detected with optical imaging techniques. (18)F decays by emitting positrons with a maximum energy of 0.635 MeV; such positrons, when travelling into tissues faster than the speed of light in the same medium, are responsible of CR emission. A detailed model of the CR spectrum considering the positron energy spectrum was developed in order to quantify the amount of light emission. The results presented in this work were obtained using a commercial optical imager equipped with charged coupled detectors (CCD). Our data open the door to optical imaging (OI) in vivo of the glucose metabolism, at least in pre-clinical research. We found that the heart and bladder can be clearly identified in the animal body reflecting the accumulation of the (18)F-FDG. Moreover, we describe two different methods based on the spectral analysis of the CR that can be used to estimate the depth of the source inside the animal. We conclude that (18)F-FDG can be employed as it is as a bimodal tracer for positron emission tomography (PET) and OI techniques. Our results are encouraging, suggesting that it could be possible to apply the proposed approach not only to beta(+) but also to pure beta(-) emitters.

In vivo optical imaging instruments are generally devoted to the acquisition of light coming from fluorescence or bioluminescence processes. Recently, an instrument was conceived with radioisotopic detection capabilities (Kodak in Vivo Multispectral System F) based on the conversion of x-rays from the phosphorus screen. The goal of this work is to demonstrate that an optical imager (IVIS 200, Xenogen Corp., Alameda, USA), designed for in vivo acquisitions of small animals in bioluminescent and fluorescent modalities, can even be employed to detect signals due to radioactive tracers. Our system is based on scintillator crystals for the conversion of high-energy rays and a collimator. No hardware modifications are required. Crystals alone permit the acquisition of photons coming from an in vivo 20 g nude mouse injected with a solution of methyl diphosphonate technetium 99 metastable (Tc99m-MDP). With scintillator crystals and collimators, a set of measurements aimed to fully characterize the system resolution was carried out. More precisely, system point spread function and modulation transfer function were measured at different source depths. Results show that system resolution is always better than 1.3 mm when the source depth is less than 10 mm. The resolution of the images obtained with radioactive tracers is comparable with the resolution achievable with dedicated techniques. Moreover, it is possible to detect both optical and nuclear tracers or bi-modal tracers with only one instrument.

Objective: 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.

Purpose: To compare two dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) techniques in terms of their ability in assessing the early antiangiogenic effect of SU11248, a novel selective multitargeted tyrosine kinase inhibitor, that exhibits direct antitumor and antiangiogenic activity via inhibition of the receptor tyrosine kinases platelet-derived growth factor receptor, vascular endothelial growth factor receptor, KIT, and FLT3. Experimental Design: A s.c. tumor model of HT29 human colon carcinoma in athymic mice was used. Two DCE-MRI techniques were used based, respectively, on macromolecular [Gd-diethylenetriaminepentaacetic acid (DTPA)-albumin] and low molecular weight (Gd-DTPA) contrast agents. The first technique provided a quantitative measurement of transendothelial permeability and fractional plasma volume, accepted surrogate markers of tumor angiogenesis. With the second technique, we quantified the initial area under the concentration-time curve, which gives information related to tumor perfusion and vascular permeability. Experiments were done before and 24 hours after a single dose administration of SU11248. Results: The early antiangiogenic effect of SU11248 was detected by DCE-MRI with macromolecular contrast agent as a 42% decrease in vascular permeability measured in the tumor rim. The effect was also detected by DCE-MRI done with Gd-DTPA as a 31% decrease in the initial area under the concentration-time curve. Histologic slices showed a statistically significant difference in mean vessel density between the treated and control groups. Conclusions: The early antiangiogenic activity of SU11248 was detected in vivo by DCE-MRI techniques using either macromolecular or low molecular weight contrast agents. Because DCE-MRI techniques with low molecular weight contrast agents can be used in clinical studies, these results could be relevant for the design of clinical trials based on new paradigms.

This preliminary study describes magnetic resonance imaging (MRI) data on the effects of sub-chronic nicotine administration in rats. Nicotine 0.4 mg/kg s.c. free base was given once a day for 4 days to Wistar adult male rats. On day 5, anaesthetized subjects were observed using an MRI tomography system. Regional cerebral blood volume (rCBV) and transversal relaxation time (T2) MRI parameters were measured. Nicotine treatment increased T2 values, with a significant effect in the cingulate cortex. A trend to increase was also observed in the prefrontal cortex and nucleus accumbens. Similarly, the effect of nicotine on rCBV was a significant increase in values compared to saline treatment. Post hoc analysis showed a significant effect of nicotine in the prefrontal cortex, cingulate cortex, mediodorsal thalamus and lateral posterior thalamus. This study showed for the first time that sub-chronic nicotine administration can induce changes in MRI pattern which are (i) similar to human MRI studies, and (ii) common to those described for markers of neuronal metabolic activation in corticolimbic brain regions known to be involved in nicotine dependence.

Contrast-enhanced ultrasound (CEUS) is an advanced approach to in vivo assessment of tumor vascularity and is being increasingly adopted in clinical oncology. It is based on 1- to 10 μm-sized gas microbubbles, which can cross the capillary beds of the lungs and are effective echo enhancers. It is known that high cell density, high transendothelial fluid exchange, and poorly functioning lymphatic circulation all provoke solid stress, which compresses vessels and drastically reduces tumor blood flow. Given their size, we supposed that the perfusion of microbubbles is affected by anatomic features of tumor vessels more than are contrast agents traditionally used in dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI). Here, we compared dynamic information obtained from CEUS and DCE-MRI on two experimental tumor models exhibiting notable differences in vessel anatomy. We found that tumors with small, flattened vessels show a much higher resistance to microbubble perfusion than to MRI contrast agents, and appear scarcely vascularized at CEUS examination, despite vessel volume adequate for normal function. Thus, whereas CEUS alone could induce incorrect diagnosis when tumors have small or collapsed vessels, integrated analysis using CEUS and DCE-MRI allows in vivo identification of tumors with a vascular profile frequently associated with malignant phenotypes.

Essential oils are currently of great importance to pharmaceutical companies, cosmetics producers and manufacturers of veterinary products. They are found in perfumes, creams, bath products, and household cleaning substances, and are used for flavouring food and drinks. It is well known that some of them act on the respiratory apparatus. The increasing interest in optical imaging techniques and the development of related technologies have made possible the investigation of the optical properties of several compounds. Luminescent properties of essential oils have not been extensively investigated. We evaluated the luminescent and fluorescent emissions of several essential oils, in order to detect them in living organisms by exploiting their optical properties. Some fluorescent emission data were high enough to be detected in dermal treatments. Consequently, we demonstrated how the fluorescent signal can be monitored for at least three hours on the skin of living mice treated with wild chamomile oil. The results encourage development of this technique to investigate the properties of drugs and cosmetics containing essential oils.