RESUMO
OBJECTIVE: Previously, the peptide sequence cNGR has been shown to home specifically to CD13/APN (aminopeptidase N) on tumor endothelium. Here, we investigated the feasibility of selective imaging of cardiac angiogenesis using the cNGR-CD13/APN system. METHODS AND RESULTS: CD13/APN induction and cNGR homing were studied in the murine myocardial infarction (MI) model. By real-time polymerase chain reaction (PCR) at 7 days after MI, CD13/APN expression was 10- to 20-fold higher in the angiogenic infarct border zone and the MI area than in non-MI areas. In vivo fluorescence microscopy confirmed specific homing of fluorophore-tagged cNGR to the border zone and MI territory at 4 and 7 days after MI with a local advantage of 2.3, but not at 1 or 14 days after MI. Tissue residence half-life was 9.1+/-0.3 hours, whereas the half-life in plasma was 15.4+/-3.4 minutes. Pulse chase experiments confirmed reversible binding of cNGR in the infarct area. Fluorescent labeled cNGR conjugates or antibodies were injected in vivo, and their distribution was studied ex vivo by 2-photon laser scanning microscopy (TPLSM). cNGR co-localized exclusively with CD13/APN and the endothelial marker CD31 on vessels. CONCLUSIONS: In cardiac angiogenesis endothelial CD13/APN is upregulated. It can be targeted specifically with cNGR conjugates. In the heart cNGR binds its endothelial target only in angiogenic areas.
Assuntos
Antígenos CD13/metabolismo , Infarto do Miocárdio/metabolismo , Neovascularização Patológica/metabolismo , Sinais Direcionadores de Proteínas , Animais , Antígenos CD13/química , Antígenos CD13/genética , Modelos Animais de Doenças , Endotélio Vascular/metabolismo , Endotélio Vascular/patologia , Regulação da Expressão Gênica/genética , Regulação da Expressão Gênica/fisiologia , Masculino , Camundongos , Microscopia de Fluorescência/métodos , Infarto do Miocárdio/genética , Infarto do Miocárdio/patologia , Nanopartículas , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Ligação ProteicaRESUMO
Thin metallic wires with an adherent hydrophilic/ lubricious polymeric coating were manufactured in a new extrusion-like procedure. This procedure is part of a novel and efficient way of assembling lubricious guide wires for intravascular interventions, such as percutaneous transluminal angioplasty. It is reported that heparin can readily be incorporated in the hydrophilic coating. A set of heparin-containing guidewire models was made and studied in detail. This showed that (i). immersion of the guide-wire models in an aqueous environment leads to release of heparin from their surface; (ii). the presence of heparin in the coating does not impede the lubricity of the coils; (iii). addition of stearic acid in the coating, next to heparin, does not influence the lubricity of the guide-wire models. Two different charges of heparin (designated heparin-low and heparin-high) were incorporated in the coating. It is discussed that release of heparin from the surface of medical devices (e.g. guide wires and catheters) is much more effective than systemic heparinization, basically because dissolved heparin molecules have a much larger probability of simply passing a medical device's surface (axial convection) rather than contacting it (radial diffusion).
