58 research outputs found
an experimental study in the rat osteoarthritis model
Background Anti-inflammatory nanoparticular compounds could represent a
strategy to diminish osteoarthritis (OA) progression. The present study was
undertaken to prove the uptake of nanoparticular dendritic polyglycerol
sulfates (dPGS) by rat-derived articular chondrocytes and to answer the
question of whether dPGS could modulate knee joint cartilage degradation in a
rat OA model and whether complications could arise. Methods dPGS uptake and
cytotoxicity was assessed in cultured primary rat-derived articular
chondrocytes. Subsequently, OA was induced in the right knee joints of 12 male
Wistar rats by medial collateral ligament and meniscus transection. Unoperated
left knees remained as controls. Six weeks post surgery six rats were either
treated daily (14 days) with 30 mg/kg dPGS (s.c.) or a similar volume of
physiological saline. Animals were analyzed clinically for gait alterations.
Explanted knee joints were studied histologically using OA scores according to
Mankin (1971), Glasson et al., (2010) and the synovitis score according to
Krenn et al., (2006). Liver, spleen and kidneys were analyzed for degenerative
changes due to dPGS accumulation. Results dPGS was taken up after 2 hours by
the chondrocytes. Whereas no significant clinical signs of OA could be
detected, at the histological level, all operated rat knee joints revealed
features of OA in the medial compartment. The values produced by both OA score
systems were lower in rats treated with dPGS compared with saline-treated
animals. Synovitis score did not significantly differ between the groups. The
analyzed organs revealed no degenerative changes. Conclusions dPGS presented
overall cyto- and biocompatibility, no accumulation in metabolizing organs and
chondroprotective properties in the osteoarthritic knee joint
Controlled Grafting Expansion Microscopy
Expansion microscopy (ExM) is a recently developed technique that allows for the resolution of structures below the diffraction limit by physically enlarging a hydrogel-embedded facsimile of the biological sample. The target structure is labeled and this label must be retained in a relative position true to the original, smaller state before expansion by linking it into the gel. However, gel formation and digestion lead to a significant loss in target-delivered label, resulting in weak signal. To overcome this problem, we have here developed an agent combining targeting, fluorescent labeling and gel linkage in a single small molecule. Similar approaches in the past have still suffered from significant loss of label. Here we show that this loss is due to insufficient surface grafting of fluorophores into the hydrogel and develop a solution by increasing the amount of target-bound monomers. Overall, we obtain a significant improvement in fluorescence signal retention and our new dye allows the resolution of nuclear pores as ring-like structures, similar to STED microscopy. We furthermore provide mechanistic insight into dye retention in ExM
AGTR1 Is Overexpressed in Neuroendocrine Neoplasms, Regulates Secretion and May Potentially Serve as a Target for Molecular Imaging and Therapy
This study identified and confirmed angiotensin II (ATII) as a strong activator of signaling in neuroendocrine neoplasm (NEN) cells. Expression analyses of the ATII receptor type 1 (AGTR1) revealed an upregulation of mRNA levels (RT-qPCR) and radioligand binding (autoradiography) in small-intestinal (n = 71) NEN tissues compared to controls (n = 25). NEN cells with high AGTR1 expression exhibited concentration-dependent calcium mobilization and chromogranin A secretion upon stimulation with ATII, blocked by AGTR1 antagonism and Gαq inhibition. ATII also stimulated serotonin secretion from BON cells. AGTR1 ligand saralasin was coupled to a near-infrared fluorescent (NIRF) dye and tested for its biodistribution in a nude mouse model bearing AGTR1-positive BON and negative QGP-1 xenograft tumors. NIRF imaging showed significantly higher uptake in BON tumors. This proof of concept establishes AGTR1 as a novel target in NEN, paving the way for translational chelator-based probes for diagnostic PET imaging and radioligand therapy
pitfalls of ester linkage
In this study, dendritic polyglycerol sulfate (dPGS) is evaluated as a
delivery platform for the anticancer, tubulin-binding drug paclitaxel (PTX).
The conjugation of PTX to dPGS is conducted via a labile ester linkage. A non-
sulfated dendritic polyglycerol (dPG) is used as a control, and the labeling
with an indocarbocyanine dye (ICC) renders multifunctional conjugates that can
be monitored by fluorescence microscopy. The conjugates are characterized by
1H NMR, UV-vis measurements, and RP-HPLC. In vitro cytotoxicity of PTX and
dendritic conjugates is evaluated using A549 and A431 cell lines, showing a
reduced cytotoxic efficacy of the conjugates compared to PTX. The study of
uptake kinetics reveals a linear, non saturable uptake in tumor cells for
dPGS-PTX-ICC, while dPG-PTX-ICC is hardly taken up. Despite the marginal
uptake of dPG-PTX-ICC, it prompts tubulin polymerization to a comparable
extent as PTX. These observations suggest a fast ester hydrolysis and
premature drug release, as confirmed by HPLC measurements in the presence of
plasma enzyme
A Cyanine‐Bridged Somatostatin Hybrid Probe for Multimodal SSTR2 Imaging in Vitro and in Vivo: Synthesis and Evaluation
Multimodal imaging probes have attracted the interest of ongoing research, for example, for the surgical removal of tumors. Modular synthesis approaches allow the construction of hybrid probes consisting of a radiotracer, a fluorophore and a targeting unit. We present the synthesis of a new asymmetric bifunctional cyanine dye that can be used as a structural and functional linker for the construction of such hybrid probes. 68Ga‐DOTATATE, a well‐characterized radiopeptide targeting the overexpressed somatostatin receptor subtype 2 (SSTR2) in neuroendocrine tumors, was labeled with our cyanine dye, thus providing additional information along with the data obtained from the radiotracer. We tested the SSTR2‐targeting and imaging properties of the resulting probe 68Ga‐DOTA‐ICC‐TATE in vitro and in a tumor xenograft mouse model. Despite the close proximity between dye and pharmacophore, we observed a high binding affinity towards SSTR2 as well as elevated uptake in SSTR2‐overexpressing tumors in the positron emission tomography (PET) scan and histological examination
Towards Probing Conformational States of Y2 Receptor Using Hyperpolarized 129Xe NMR
G protein-coupled receptors can adopt many different conformational states, each of them exhibiting different restraints towards downstream signaling pathways. One promising strategy to identify and quantify this conformational landscape is to introduce a cysteine at a receptor site sensitive to different states and label this cysteine with a probe for detection. Here, the application of NMR of hyperpolarized 129Xe for the detection of the conformational states of human neuropeptide Y2 receptor is introduced. The xenon trapping cage molecule cryptophane-A attached to a cysteine in extracellular loop 2 of the receptor facilitates chemical exchange saturation transfer experiments without and in the presence of native ligand neuropeptide Y. High-quality spectra indicative of structural states of the receptor–cage conjugate were obtained. Specifically, five signals could be assigned to the conjugate in the apo form. After the addition of NPY, one additional signal and subtle modifications in the persisting signals could be detected. The correlation of the spectroscopic signals and structural states was achieved with molecular dynamics simulations, suggesting frequent contact between the xenon trapping cage and the receptor surface but a preferred interaction with the bound ligand
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Design of Polymer-Embedded Heterogeneous Fenton Catalysts for the Conversion of Organic Trace Compounds
Advanced oxidation processes are the main way to remove persistent organic trace compounds from water. For these processes, heterogeneous Fenton catalysts with low iron leaching and high catalytic activity are required. Here, the preparation of such catalysts consisting of silica-supported iron oxide (Fe2O3/SiOx) embedded in thermoplastic polymers is presented. The iron oxide catalysts are prepared by a facile sol–gel procedure followed by thermal annealing (calcination). These materials are mixed in a melt compounding process with modified polypropylenes to stabilize the Fe2O3 catalytic centers and to further reduce the iron leaching. The catalytic activity of the composites is analyzed by means of the Reactive Black 5 (RB5) assay, as well as by the conversion of phenol which is used as an example of an organic trace compound. It is demonstrated that embedding of silica-supported iron oxide in modified polypropylene turns the reaction order from pseudo-first order (found for Fe2O3/SiOx catalysts), which represents a mainly homogeneous Fenton reaction, to pseudo-zeroth order in the polymer composites, indicating a mainly heterogeneous, surface-diffusion-controlled process
Optical Imaging
Optical Coherence Tomography (OCT)We describe the fundamental concept of optical coherence tomography (OCT) and discuss the two main working principles time domain OCT and frequency domain OCT. Then, we review extended functionalities including spectrally and polarization-resolved OCT as well as Doppler-OCT and show concepts for contrast enhancement. Based on these fundamentals, we demonstrate the potential of OCT for small animal imaging on the basis of exemplary studies on retinal imaging and lung imaging.Optoacoustic ImagingThis chapter deals with the fascinating topic of optoacoustic imaging, a recent powerful addition to the arsenal of in vivo functional and molecular small animal imaging. Due to its hybrid nature, involving optical excitation and ultrasonic detection, optoacoustics overcomes the imaging depth limitations of optical microscopy related to light scattering in living tissues while further benefiting from the compelling advantages of optical contrast. To this end, optoacoustic imaging has been shown capable of delivering multiple types of imaging contrast (structural, functional, kinetic, molecular) within a single imaging modality. It can further deliver images with high spatiotemporal resolution that rivals performance of other well-established whole-body imaging modalities. As such, optoacoustics can play a vital role in biomedical research, from early disease detection and monitoring of dynamic phenomena noninvasively to accelerating drug discovery.Optical ProbesThis chapter is devoted to the properties and application of fluorescence dyes as probes for optical imaging. A variety of agents have been described to date, including nontargeting dyes, vascular agents, targeted conjugates, activatable dyes, and sensing probes. The major classes encompass polymethine dyes and xanthenes dyes, both of which are commercially available in broad variations. Addressing the purpose of optical animal imaging, the most relevant parameters to apply such probes are discussed, thereby supporting the reader in choosing reasonable imaging probes and in preparing bioconjugates for his studies
Multidisciplinary Ophthalmic Imaging In Vivo Imaging of a New Indocyanine Green Micelle Formulation in an Animal Model of Laser-Induced Choroidal Neovascularization
METHODS. The ICG was formulated with the nonionic solubilizer and emulsifying agent Kolliphor HS 15 to create ICG/HS 15 to improve the chemical stability and fluorescence efficacy. In vivo imaging was performed in rats that had undergone laser photocoagulation. Retinal uptake and fluorescence intensity of ICG and ICG/HS 15 were compared following intravenous injection of 3 dosages (0.05, 0.1, and 0.15 mg/kg body weight) at 7, 14, and 21 days following laser treatment. Postmortem analysis included histology with frozen sections and flat mounts. RESULTS. Immediately following injection of ICG or ICG/HS 15, a strong fluorescence was visible in the retinal vasculature and at the site of laser lesions. Pixel intensity was higher for ICG/HS 15 compared to conventional ICG at 8 minutes after injection for all different injection days and dosages. Over time, a continuous decrease of the fluorescent signal was observed for up to 60 minutes to baseline level. Flow cytometry data showed an increased uptake of micellar dye of macrophages and endothelial cells. Histology revealed an accumulation of the micellar dye within the laser lesion. CONCLUSIONS. Micelle formulated ICG can be visualized in the retinal vasculature and laserinduced CNV in vivo and ex vivo. Micellar ICG/HS 15 showed in vivo stronger signal intensity when compared to ICG for all tested dosages. Following further investigations, ICG/HS 15 may be evaluated in patients with retinal and choroidal diseases for more refined diagnosis
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