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      Early and late pulmonary effects of nebulized LPS in mice: An acute lung injury model

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          Abstract

          Background and objective

          Acute respiratory distress syndrome (ARDS) has a high mortality rate of 35–46% depending on its severity. Animal models are crucial to better understand the pathophysiology of diseases, including ARDS. This study presents a feasible animal model of acute lung injury (ALI) using nebulized lipopolysaccharide (LPS) in a non-invasive approach, focusing on its short and long-term effects.

          Methods

          Mice received nebulized LPS or vehicle only (control group). Blood, BALF and lung tissue were collected 24 hours (LPS 24h) or 5 weeks (LPS 5w) after the nebulized LPS-induced lung injury. Inflammatory cytokines were assessed in the blood serum, BALF and lung tissue. Stereological analyses and remodeling changes were assessed by histology and immunohistochemistry at the specified time points.

          Results

          The LPS 24h group showed increased pro-inflammatory cytokine levels, intense cell influx, increased total septal volume, septal thickening and decreased surface density of the alveolar septa. The LPS 5w group showed persistent lung inflammation, septal thickening, increased total lung volume, accentuated collagen deposition, especially of collagen type I, and decreased MMP-2 protein expression.

          Conclusion

          We present a feasible, reproducible and non-invasive nebulized-LPS animal model that allows the assessment of both the acute and late phases of acute lung injury. The presence of lung remodeling with collagen deposition after 5 weeks makes it useful to study the pathophysiology, complications, and possible therapeutic intervention studies that aim to understand and reduce pulmonary fibrosis in the late phases of ALI.

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          Most cited references27

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          An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals.

          Acute lung injury (ALI) is well defined in humans, but there is no agreement as to the main features of acute lung injury in animal models. A Committee was organized to determine the main features that characterize ALI in animal models and to identify the most relevant methods to assess these features. We used a Delphi approach in which a series of questionnaires were distributed to a panel of experts in experimental lung injury. The Committee concluded that the main features of experimental ALI include histological evidence of tissue injury, alteration of the alveolar capillary barrier, presence of an inflammatory response, and evidence of physiological dysfunction; they recommended that, to determine if ALI has occurred, at least three of these four main features of ALI should be present. The Committee also identified key "very relevant" and "somewhat relevant" measurements for each of the main features of ALI and recommended the use of least one "very relevant" measurement and preferably one or two additional separate measurements to determine if a main feature of ALI is present. Finally, the Committee emphasized that not all of the measurements listed can or should be performed in every study, and that measurements not included in the list are by no means "irrelevant." Our list of features and measurements of ALI is intended as a guide for investigators, and ultimately investigators should choose the particular measurements that best suit the experimental questions being addressed as well as take into consideration any unique aspects of the experimental design.
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            Resolution of inflammation: an integrated view

            Resolution of inflammation is a coordinated and active process aimed at restoration of tissue integrity and function. This review integrates the key molecular and cellular mechanisms of resolution. We describe how abrogation of chemokine signalling blocks continued neutrophil tissue infiltration and how apoptotic neutrophils attract monocytes and macrophages to induce their clearance. Uptake of apoptotic neutrophils by macrophages reprograms macrophages towards a resolving phenotype, a key event to restore tissue homeostasis. Finally, we highlight the therapeutic potential that derives from understanding the mechanisms of resolution.
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              Animal models of fibrotic lung disease.

              Interstitial lung fibrosis can develop as a consequence of occupational or medical exposure, as a result of genetic defects, and after trauma or acute lung injury leading to fibroproliferative acute respiratory distress syndrome, or it can develop in an idiopathic manner. The pathogenesis of each form of lung fibrosis remains poorly understood. They each result in a progressive loss of lung function with increasing dyspnea, and most forms ultimately result in mortality. To better understand the pathogenesis of lung fibrotic disorders, multiple animal models have been developed. This review summarizes the common and emerging models of lung fibrosis to highlight their usefulness in understanding the cell-cell and soluble mediator interactions that drive fibrotic responses. Recent advances have allowed for the development of models to study targeted injuries of Type II alveolar epithelial cells, fibroblastic autonomous effects, and targeted genetic defects. Repetitive dosing in some models has more closely mimicked the pathology of human fibrotic lung disease. We also have a much better understanding of the fact that the aged lung has increased susceptibility to fibrosis. Each of the models reviewed in this report offers a powerful tool for studying some aspect of fibrotic lung disease.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: Project administrationRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: Validation
                Role: Investigation
                Role: InvestigationRole: Visualization
                Role: Investigation
                Role: Formal analysisRole: Investigation
                Role: InvestigationRole: MethodologyRole: Validation
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: MethodologyRole: Resources
                Role: ConceptualizationRole: Funding acquisitionRole: Resources
                Role: ConceptualizationRole: MethodologyRole: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                27 September 2017
                2017
                : 12
                : 9
                : e0185474
                Affiliations
                [1 ] Laboratory of Experimental Air Pollution (LIM05), University of Sao Paulo—School of Medicine, São Paulo, São Paulo, Brazil
                [2 ] Biochemistry Laboratory, University of Sao Paulo–School of Pharmaceutical Sciences, São Paulo, São Paulo, Brazil
                [3 ] Laboratory of Clinical Immunology and Allergy (LIM60), University of Sao Paulo—School of Medicine, São Paulo, São Paulo, Brazil
                [4 ] Department of Pathology, Case Western Reserve University, Cleveland, Ohio, United States of America
                Hospital for Sick Children, CANADA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-7848-2092
                Article
                PONE-D-17-18837
                10.1371/journal.pone.0185474
                5617199
                28953963
                4721a0ab-5327-46c5-97af-ff63704e6a5f
                © 2017 de Souza Xavier Costa et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 16 May 2017
                : 13 September 2017
                Page count
                Figures: 6, Tables: 0, Pages: 16
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100001807, Fundação de Amparo à Pesquisa do Estado de São Paulo;
                Award ID: 2008/57717-6
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100003593, Conselho Nacional de Desenvolvimento Científico e Tecnológico;
                Award ID: 573813/2008-6
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001807, Fundação de Amparo à Pesquisa do Estado de São Paulo;
                Award ID: 2012/23334-9
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100003593, Conselho Nacional de Desenvolvimento Científico e Tecnológico;
                Award ID: 479679/2012-6
                Award Recipient :
                This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) - 2008/57717-6 - PHNS; Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) - 2012/23334-9 - NSXC; Conselho Nacional de Desenvolvimento Científico e Tecnológico - 573813/2008-6 - PHNS; Conselho Nacional de Desenvolvimento Científico e Tecnológico - 479679/2012-6 - LFFS; Conselho Nacional de Desenvolvimento Científico e Tecnológico: http://cnpq.br/ and Fundação de Amparo à Pesquisa do Estado de São Paulo: http://www.fapesp.br/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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