RESUMEN
Today, it is increasingly recognized that air pollution hurts human health. Consequently, efficient mitigation strategies need to be implemented for substantial environmental and health co-benefits. A valid approach to reducing the air pollution effects on the environment and human health is proposed. Specific guidelines have been elucidated by differentiating them on the base of the final stakeholders (citizens, enterprises, and public authorities), of the emission sources (transport, household energy, industry, and energy generation sector, agriculture, and shipping area), and of the field of implementation (urban and extra-urban context). This paper can provide useful information for governments for the implementation of a strategic plan focused on emphasizing multi-pollutant emission reductions and overall air pollution-related risk.
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Contaminantes Atmosféricos/análisis , Contaminación del Aire/análisis , Agricultura , HumanosRESUMEN
The use of 3D cancer models will have both ethical and economic impact in drug screening and development, to promote the reduction of the animals employed in preclinical studies. Nevertheless, to be effective, such cancer surrogates must preserve the physiological relevance of the in vivo models in order to provide realistic information on drugs' efficacy. To figure out the role of the architecture and composition of 3D cancer models on their tumor-mimicking capability, here we studied the efficacy of doxorubicin (DOX), a well-known anticancer molecule in two different 3D cancer models: our 3D breast cancer microtissue (3D-µTP) versus the golden standard represented by spheroid model (sph). Both models were obtained by using cancer associated fibroblast (CAF) and breast cancer cells (MCF-7) as cellular component. Unlike spheroid model, 3D-µTP was engineered in order to induce the production of endogenous extracellular matrix by CAF. 3D-µTP have been compared to spheroid in mono- (MCF-7 alone) and co-culture (MCF-7/CAF), after the treatment with DOX in order to study cytotoxicity effect, diffusional transport and expression of proteins related to cancer progression. Compared to the spheroid model, 3D-µTP showed higher diffusion coefficient of DOX and lower cell viability. Also, the expression of some tumoral biomarkers related to cell junctions were different in the two models. STATEMENTS OF SIGNIFICANCE: Cancer biology has made progress in unraveling the mechanism of cancer progression, anyway the most of the results are still obtained by 2D cell cultures or animal models, that do not faithfully copycat the tumor microenvironment. The lack of correlation between preclinical models and in vivo organisms negatively influences the clinical efficacy of chemotherapeutic drugs. Consequently, even if a huge amount of new drugs has been developed in the last decades, still people are dying because of cancer. Pharmaceutical companies are interested in 3D tumor model as valid alternative in drug screening in preclinical studies. However, a 3D tumor model that completely mimics tumor heterogeneity is still far to achieve. In our work we compare 3D human breast cancer microtissues and spheroids in terms of response to doxorubicin and drug diffusion. We believe that our results are interesting because they highlight the potential role of the proposed tumor model in the attempts to improve efficacy tests.
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Neoplasias de la Mama , Doxorrubicina/farmacología , Modelos Biológicos , Esferoides Celulares , Microambiente Tumoral , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Femenino , Humanos , Células MCF-7 , Esferoides Celulares/metabolismo , Esferoides Celulares/patologíaRESUMEN
Therapeutic approaches based on nanomedicine have garnered great attention in cancer research. In vitro biological models that better mimic in vivo conditions are crucial tools to more accurately predict their therapeutic efficacy in vivo. In this work, a new 3D breast cancer microtissue has been developed to recapitulate the complexity of the tumor microenvironment and to test its efficacy as screening platform for drug delivery systems. The proposed 3D cancer model presents human breast adenocarcinoma cells and cancer-associated fibroblasts embedded in their own ECM, thus showing several features of an in vivo tumor, such as overexpression of metallo-proteinases (MMPs). After demonstrating at molecular and protein level the MMP2 overexpression in such tumor microtissues, we used them to test a recently validated formulation of endogenous MMP2-responsive nanoparticles (NP). The presence of the MMP2-sensitive linker allows doxorubicin release from NP only upon specific enzymatic cleavage of the peptide. The same NP without the MMP-sensitive linker and healthy breast microtissues were also produced to demonstrate NP specificity and selectivity. Cell viability after NP treatment confirmed that controlled drug delivery is achieved only in 3D tumor microtissues suggesting that the validation of therapeutic strategies in such 3D tumor model could predict human response. STATEMENT OF SIGNIFICANCE: A major issue of modern cancer research is the development of accurate and predictive experimental models of human tumors consistent with tumor microenvironment and applicable as screening platforms for novel therapeutic strategies. In this work, we developed and validated a new 3D microtissue model of human breast tumor as a testing platform of anti-cancer drug delivery systems. To this aim, biodegradable nanoparticles responsive to physiological changes specifically occurring in tumor microenvironment were used. Our findings clearly demonstrate that the breast tumor microtissue well recapitulates in vivo physiological features of tumor tissue and elicits a specific response to microenvironmentally-responsive nanoparticles compared to healthy tissue. We believe this study is of particular interest for cancer research and paves the way to exploit tumor microtissues for several testing purposes.
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Neoplasias de la Mama , Doxorrubicina , Sistemas de Liberación de Medicamentos/métodos , Metaloproteinasa 2 de la Matriz/metabolismo , Nanopartículas/química , Proteínas de Neoplasias/metabolismo , Microambiente Tumoral/efectos de los fármacos , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Doxorrubicina/química , Doxorrubicina/farmacología , Femenino , Humanos , Células MCF-7RESUMEN
We fabricated three-dimensional microtissues with the aim to replicate in vitro the composition and the functionalities of the tumor microenvironment. By arranging either normal fibroblasts (NF) or cancer-activated fibroblasts (CAF) in two different three dimensional (3D) configurations, two kinds of micromodules were produced: spheroids and microtissues. Spheroids were obtained by means of the traditional cell aggregation technique resulting in a 3D model characterized by high cell density and low amount of extracellular proteins. The microtissues were obtained by culturing cells into porous gelatin microscaffolds. In this latter configuration, cells assembled an intricate network of collagen, fibronectin and hyaluronic acid. We investigated the biophysical properties of both 3D models in terms of cell growth, metabolic activity, texture and composition of the extracellular matrix (via histological analysis and multiphoton imaging) and cell mechanical properties (via Particle Tracking Microrheology). In the spheroid models such biophysical properties remained unchanged regardless to the cell type used. In contrast, normal-microtissues and cancer-activated-microtissues displayed marked differences. CAF-microtissues possessed higher proliferation rate, superior contraction capability, different micro-rheological properties and an extracellular matrix richer in collagen fibronectin and hyaluronic acid. At last, multiphoton investigation revealed differences in the collagen network architecture. Taken together, these results suggested that despite to cell spheroids, microtissues better recapitulate the important differences existing in vivo between normal and cancer-activated stroma representing a more suitable system to mimic in vitro the stromal element of the tumor tissues. STATEMENT OF SIGNIFICANCE: This work concerns the engineering of tumor tissue in vitro. Tumor models serve as biological equivalent to study pathologic progression and to screen or validate the drugs efficacy. Tumor tissue is composed by malignant cells surviving in a microenvironment, or stroma. Stroma plays a pivotal role in cancer progression. Current in vitro models, i.e. spheroids, can't replicate the phenomena related to the tumor stroma remodeling. For this reason, to better replicate the tumor physiology in vitro that include functional and morphological changes, a novel 3D cancer model is proposed.
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Microambiente Celular , Neoplasias/patología , Recuento de Células , Núcleo Celular/metabolismo , Colágeno/metabolismo , Matriz Extracelular/metabolismo , Fibroblastos/patología , Fibronectinas/metabolismo , Técnica del Anticuerpo Fluorescente , Humanos , Ácido Hialurónico/metabolismo , Consumo de Oxígeno , Reología , Esferoides Celulares/patología , Células del Estroma/patología , Factores de TiempoRESUMEN
In this work, a new model of breast cancer is proposed featuring both epithelial and stromal tissues arranged on a microfluidic chip. The main task of the work is the in vitro replication of the stromal activation during tumor epithelial invasion. The activation of tumor stroma and its morphological/compositional changes play a key role in tumor progression. Despite emerging evidences, to date the activation of tumor stroma in vitro has not been achieved yet. The tumor-on-chip proposed in this work is built in order to replicate the features of its native counterpart: multicellularity (tumor epithelial cell and stromal cell); 3D engineered stroma compartment composed of cell-assembled extracellular matrix (ECM); reliable 3D tumor architecture. During tumor epithelial invasion the stroma displayed an activation process at both cellular and ECM level. Similarly of what repeated in vivo, ECM remodeling is found in terms of hyaluronic acid and fibronectin overexpression in the stroma compartment. Furthermore, the cell-assembled ECM featuring the stromal tissue, allowed on-line monitoring of collagen remodeling during stroma activation process via real time multiphoton microscopy. Also, trafficking of macromolecules within the stromal compartment has been monitored in real time.