RESUMO
Parkinson's disease (PD) is a complex and multifaceted neurodegenerative disorder that results from multiple environmental factors and multicellular interactions. Although several PD neuropathologies have been identified and described, the thorough understanding of PD pathophysiology and research has been largely limited by the absence of reliablein vitromodels that truly recapitulate PD microenvironments. Here, we propose a neuroimmune co-culture system that models PD neuropathologies by combining relevant multicellular interactions with environments that mimic the brain. This system is composed of: (i) 3D bioprinted cultures of mature human dopaminergic (DA) neurons grown on extracellular matrix (ECM)-derived scaffolds doped with electroconductive nanostructures, and (ii) a direct co-culture of human astrocytes and differentiated monocytes that models neuroinflammatory responses. When co-cultured in a transwell format, these two compartments recreate relevant multicellular environments that model PD pathologies after exposure to the neurotoxin A53Tα-synuclein. With immunofluorescent staining and gene expression analyses, we show that functional and mature DA 3D networks are generated within our ECM-derived scaffolds with superior performance to standard 2D cultures. Moreover, by analyzing cytokine secretion, cell surface markers, and gene expression, we define a human monocyte differentiation scheme that allows the appearance of both monocyte-derived macrophages and dendritic cell phenotypes, as well as their optimal co-culture ratios with human astrocytes to recreate synergistic neuroinflammatory responses. We show that the combined response of both compartments to A53Tα-synuclein stimulates the formation of intracellularα-synuclein aggregates, induces progressive mitochondrial dysfunction and reactive oxygen species production, downregulates the expression of synaptic, DA, and mitophagy-related genes, and promotes the initiation of apoptotic processes within the DA networks. Most importantly, these intracellular pathologies were comparable or superior to those generated with a rotenone-stimulated 2D control that represents the current standard forin vitroPD models and showed increased resilience towards these neurotoxic insults, allowing the study of disease progression over longer time periods than current models. Taken together, these results position the proposed model as a superior alternative to current 2D models for generating PD-related pathologiesin vitro.
Assuntos
Doença de Parkinson , Humanos , Doença de Parkinson/metabolismo , Doença de Parkinson/patologia , Técnicas de Cocultura , alfa-Sinucleína/metabolismo , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/patologia , Macrófagos , InflamaçãoRESUMO
Las construcciones y deconstrucciones de la práctica profesional en psicología, son el resultado del ejercicio riguroso académico que el programa a través del comité curricular, comité de prácticas y las distintas reflexiones de los docentes, graduados, cooperadores en las agencias, estudiantes y directivos realizan permanentemente con el fin de fundamentar las acciones desde lo social a través de los distintos escenarios en los que el estudiante que se encuentra en formación encontrará una vez inicie este camino hacia la "praxis". Comprendemos y asentimos que el inicio de la práctica implica una responsabilidad con la sociedad, la institución universitaria y el aprendiz que se lanza al mundo a ser y hacer frente a lo que sabe y como lo sabe
The constructions and deconstructions of professional practice in psychology are the result of the rigorous academic exercise that the program through the curricular committee, practice committee and the different reflections of teachers, graduates, cooperators in agencies, students and managers permanently carry out in order to base actions from the social point of view through the different scenarios in which the student who is in training will find once he or she begins this path towards "praxis". We understand and agree that the beginning of the practice implies a responsibility with society, the university institution and the apprentice who launches himself into the world to be and face what he knows and how he knows it