A Trimodal Imaging Platform for Tracking Viable Transplanted Pancreatic Islets In Vivo: F-19 MR, Fluorescence, and Bioluminescence Imaging.

PURPOSE: Combining specific and quantitative F-19 magnetic resonance imaging (MRI) with sensitive and convenient optical imaging provides complementary information about the distribution and viability of transplanted pancreatic islet grafts. In this study, pancreatic islets (PIs) were labeled with positively charged multimodal nanoparticles based on poly(lactic-co-glycolic acid) (PLGA-NPs) with encapsulated perfluoro-15-crown-5-ether and the near-infrared fluorescent dye indocyanine green. PROCEDURES: One thousand and three thousand bioluminescent PIs were transplanted into subcutaneous artificial scaffolds, which served as an alternative transplant site. The grafts were monitored using in vivo F-19 MR, fluorescence, and bioluminescence imaging in healthy rats for 2 weeks. RESULTS: Transplanted PIs were unambiguously localized in the scaffolds by F-19 MRI throughout the whole experiment. Fluorescence was detected in the first 4 days after transplantation only. Importantly, in vivo bioluminescence correlated with the F-19 MRI signal. CONCLUSIONS: We developed a trimodal imaging platform for in vivo examination of transplanted PIs. Fluorescence imaging revealed instability of the fluorescent dye and its limited applicability for longitudinal in vivo studies. A correlation between the bioluminescence signal and the F-19 MRI signal indicated the fast clearance of PLGA-NPs from the transplantation site after cell death, which addresses a major issue with intracellular imaging labels. Therefore, the proposed PLGA-NP platform is reliable for reflecting the status of transplanted PIs in vivo.

Mannan-based conjugates as a multimodal imaging platform for lymph nodes.

We show that mannan-based conjugates possess exceptional features for multimodal imaging because of their biocompatibility, biodegradability and self-targeting properties. Two new mannan conjugates, containing a gadolinium complex and a fluorescent probe, one based only on polysaccharide and the other one comprising polysaccharide with poly(2-methyl-2-oxazoline) grafts, were prepared and simultaneously visualized in vitro and in vivo by magnetic resonance and fluorescence imaging. The synthesis of these mannan-based complexes was based on alkylation with allyl bromide or grafting with poly(2-methyl-2-oxazoline) chains, followed by a thiol-ene click reaction with cysteamine to introduce primary amino groups into their structure. Finally, the obtained conjugates were functionalized with contrast labels using the corresponding N-hydroxysuccinimide esters. When used to detect lymph nodes, the polymers showed better imaging properties than a commercially available contrast agent.

Ln(iii)-complexes of a DOTA analogue with an ethylenediamine pendant arm as pH-responsive PARACEST contrast agents.

A novel macrocyclic DO3A derivative containing a linear diamine pendant arm, H3do3aNN, was prepared and its protonation and complexation properties were studied by means of potentiometry. It determined ligand consecutive protonation constants log K(An) = 12.62, 10.28, 9.67, 8.30, 3.30 and 1.58 and stability constants of selected lanthanide (Eu(iii), Yb(iii)) complexes log K(EuL) = 23.16 and log KYbL = 22.76. The complexes could be protonated on the pendant amino group(s) with log K(HLM) ≈ 5.6 and log K(H2LM) ≈ 4.8. Solution structures of both complexes were studied by NMR spectroscopy. The study revealed that the complex species exist exclusively in the form of twisted-square-antiprismatic (TSA) isomers. The complexes show significant pH dependence of the Chemical Exchange Saturation Transfer (CEST) between their amino groups and the bulk water molecules in the pH range of 5-8. Thus, the pH dependence of the magnetization transfer ratio of CEST signals can be used for pH determination using magnetic resonance imaging techniques in a pH range relevant for in vivo conditions.

Effect of mesenchymal stem cells on the vascularization of the artificial site for islet transplantation in rats.

An adequate vascularization of the artificially created cavity is crucial for subsequent transplantation of isolated pancreatic islets. In a reported study, dynamic contrast-enhanced magnetic resonance imaging was used to assess the effect of mesenchymal stem cells on neoangiogenesis within connective tissue surrounding an implantable biocompatible device. The signal increase detected after injection of magnetic resonance contrast agent in each target region was considered to be an effect of contrast agent, which was related to the blood supply. To minimize the influence of variability in contrast agent application, all outcomes measured in the implanted devices were normalized to the signal intensity of kidney tissue. When supported by mesenchymal stem cells, the mean signal increase intra-abdominally was 42%, 41%, and 64% and within subcutaneously implanted devices was 23%, 54%, and 52% of that measured in kidney.