RESUMEN
The insertion of a stent in diseased arteries is a common endovascular procedure that can be compromised by the development of short- and long-term inflammatory responses leading to restenosis and thrombosis, respectively. While treatment with drugs, either systemic or localized, has decreased the incidence of restenosis and thrombosis these complications persist and are associated with a high mortality in those that present with stent thrombosis. We reasoned that if stents could be made to undergo accelerated endothelialization in the deployed region, then such an approach would further decrease the occurrence of stent thrombosis and restenosis thereby improving clinical outcomes. Toward that objective, the first step necessitated efficient capture of progenitor stem cells, which eventually would become the new endothelium. To achieve this objective, we engineered intrinsic ferromagnetism within nonmagnetizable, biodegradable magnesium (Mg) bare metal stents. Mg stents were coated with biodegradable polylactide (PLA) polymer embedding magnetizable iron-platinum (FePt) alloy nanoparticles, nanomagnetic particles, nMags, which increased the surface area and hence magnetization of the stent. nMags uniformly distributed on stents enabled capture, under flow, up to 50 mL/min, of systemically injected iron-oxide-labeled (IO-labeled) progenitor stem cells. Critical parameters enhancing capture efficiency were optimized, and we demonstrated the generality of the approach by showing that nMag-coated stents can capture different cell types. Our work is a potential paradigm shift in engineering stents because implants are rendered as tissue in the body, and this "natural stealthiness" reduces or eliminates issues associated with pro-inflammatory immune responses postimplantation.
Asunto(s)
Materiales Biocompatibles Revestidos/química , Reestenosis Coronaria/prevención & control , Magnesio/química , Imanes/química , Poliésteres/química , Stents , Trombosis/prevención & control , Aleaciones/química , Células Endoteliales de la Vena Umbilical Humana , Humanos , Hierro/química , Nanopartículas del Metal/química , Nanopartículas del Metal/ultraestructura , Platino (Metal)/química , Células Madre/citología , Stents/efectos adversosRESUMEN
The diffusive transfer, or paracrine delivery, of chemical factors during the interaction of an emitting cell and a receiving cell is a ubiquitous cellular process that facilitates information exchange between the cells an/or to bystander cells. In the cellular immune response this exchange governs the magnitude and breadth of killing of cellular targets, inflammation or tolerance. Paracrine delivery is examined here by solving the the steady-state diffusion equation for the concentration field surrounding two intensely interacting, equi-sized cells on which surface kinetics limits the rates of factor emission and absorption. These chemical factors may be cytokines, such as Interlukins and Interferons, but the results are presented in a generic form so as to be applicable to any chemical factor and/or cell-type interaction. In addition to providing overall transfer rates and transfer efficiencies, the results also indicate that when the receiving cell is naïve, with few factor receptors on its surface, there may be a significant accumulation of factor in the synaptic region between the cells with a consequent release of factor to the medium where it can signal bystander cells. This factor accumulation may play a critical role in activating a naïve receiving cell. As the receiving cell activates and becomes more absorbent, the factor accumulation diminishes, as does potential bystander signaling.
RESUMEN
Conventional immunosuppressive drug delivery requires high systemic drug levels to provide therapeutic benefit, but frequently results in toxic side effects. Novel drug delivery methods, such as FDA-approved poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs), are promising drug delivery platforms to reduce drug doses and minimize toxicity. Using murine models of skin transplantation, we investigated whether PLGA NPs would effectively deliver mycophenolic acid (MPA), a common clinical immunosuppressant, and avoid the toxicity of conventional drug delivery. We found that intermittent treatment with NPs encapsulated with MPA (NP-MPA) resulted in a significant extension of allograft survival than intermittent conventional MPA treatment even though the concentration of MPA within NP-MPA was a 1000-fold lower than conventional drug. Importantly, recipients who were administered NP-MPA intermittently avoided drug toxicity, whereas those treated with daily conventional drug manifested cytopenias. Dendritic cells (DCs) endocytosed NP-MPA to upregulate programmed death ligand-1 (PD-L1) and displayed a decreased ability to prime alloreactive T cells. Importantly, the ability of NP-MPA to promote allograft survival was partly PD-L1 dependent. Collectively, this study indicates that NPs are potent drug delivery tools that extend allograft survival without drug toxicity.
Asunto(s)
Antígeno B7-H1/metabolismo , Células Dendríticas/efectos de los fármacos , Sistemas de Liberación de Medicamentos , Supervivencia de Injerto/efectos de los fármacos , Ácido Micofenólico/administración & dosificación , Nanopartículas/química , Enfermedades de la Piel/terapia , Animales , Antibióticos Antineoplásicos/administración & dosificación , Antibióticos Antineoplásicos/farmacocinética , Células Cultivadas , Terapia Combinada , Células Dendríticas/metabolismo , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Ácido Láctico/química , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ácido Micofenólico/farmacocinética , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Enfermedades de la Piel/inmunología , Enfermedades de la Piel/mortalidad , Trasplante de Piel , Tasa de Supervivencia , Distribución Tisular , Trasplante HomólogoRESUMEN
This pilot study examines noninvasive MR monitoring of tissue-engineered vascular grafts (TEVGs) in vivo using cells labeled with iron oxide nanoparticles. Human aortic smooth muscle cells (hASMCs) were labeled with ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles. The labeled hASMCs, along with human aortic endothelial cells, were incorporated into eight TEVGs and were then surgically implanted as aortic interposition grafts in a C.B-17 SCID/bg mouse host. USPIO-labeled hASMCs persisted in the grafts throughout a 3 wk observation period and allowed noninvasive MR imaging of the human TEVGs for real-time, serial monitoring of hASMC retention. This study demonstrates the feasibility of applying noninvasive imaging techniques for evaluation of in vivo TEVG performance.
Asunto(s)
Aorta/citología , Prótesis Vascular , Medios de Contraste/farmacología , Hierro/farmacología , Angiografía por Resonancia Magnética/métodos , Músculo Liso Vascular/citología , Miocitos del Músculo Liso/citología , Nanopartículas , Óxidos/farmacología , Ingeniería de Tejidos/métodos , Animales , Dextranos , Óxido Ferrosoférrico , Humanos , Nanopartículas de Magnetita , Ratones , Ratones SCIDRESUMEN
While activated T cells are known to have enhanced biological responses to antigen stimulation, the biophysical basis of this increased sensitivity remains unknown. Here, we show that, on activated T cells, the TCR avidity for peptide-MHC complexes is 20- to 50-fold higher than the TCR avidity of naive T cells. This increased avidity for peptide-MHC depends on TCR reorganization and is sensitive to the cholesterol content of the T cell membrane. Analysis of the binding data indicates the enhanced avidity is due to increases in cross-linking of TCR on activated T cells. Activation-induced membrane (AIM) changes in TCR avidity represent a previously unrecognized means of increasing the sensitivity of activated T cells to small amounts of antigen in the periphery.