RESUMO
Although empyema affects more than 65,000 people each year in the United States and in the United Kingdom, there are limited data on the pathogenesis of pleural infection. We investigated the pathogenesis of empyema using animal and cell culture models of Streptococcus pneumoniae infection. The pathological processes during the development of empyema associated with murine pneumonia due to S. pneumoniae (strain D39) were investigated. Lungs were examined using histology, and pleural fluid and blood bacterial colony-forming units, cytokine levels, and cellular infiltrate were determined over time. Bacterial migration across mesothelial monolayers was investigated using cell culture techniques, flow cytometry, and confocal microscopy. After intranasal inoculation with 10(7) S. pneumoniae D39 strain, mice developed pneumonia associated with rapid bacterial invasion of the pleural space; raised intrapleural IL-8, VEGF, MCP-1, and TNF-α levels; and caused significant intrapleural neutrophilia followed by the development of fibrinous pleural adhesions. Bacterial clearance from the pleural space was poor, and in vitro assays demonstrated that S. pneumoniae crossed mesothelial layers by translocation through cells rather than by a paracellular route. This study describes key events during the development of S. pneumoniae empyema using a novel murine model of pneumonia-associated empyema that closely mimics human disease. The model allows for future assessment of molecular mechanisms involved in the development of empyema and evaluation of potential new therapies. The data suggest that transmigration of bacteria through mesothelial cells could be important in empyema development. Furthermore, upon entry the pleural cavity offers a protected compartment for the bacteria.
Assuntos
Modelos Animais de Doenças , Empiema/fisiopatologia , Pneumopatias/microbiologia , Pleura/microbiologia , Doenças Pleurais/microbiologia , Streptococcus pneumoniae/patogenicidade , Animais , Empiema/microbiologia , CamundongosRESUMO
In humans, the greater omentum is a fatty peritoneal fold that extends from the greater curvature of the stomach to cover most abdominal organs. It performs many functions, which include acting as a reservoir of resident peritoneal inflammatory cells, a storage site for lipid, and a regulator of fluid exchange in and out of the peritoneal cavity. Most importantly, the omentum readily adheres to areas of inflammation and peritoneal damage, often leading to adhesion formation. Despite its clinical importance, the omentum remains an understudied organ, and discrepancies exist as to its exact morphology. This study uses a combination of phase contrast microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) to elucidate the structure of the greater omentum of both human and mouse and determine whether it possesses a typical surface mesothelial cell lining similar to other serosa. Results indicated that both human and murine omenta were of similar structure and composed of two distinct types of tissue, one adipose-rich and the other translucent and membranous. The adipose-rich regions were well-vascularised and covered by a continuous mesothelial cell layer except at the sites of milky spots. In contrast, translucent areas were poorly vascularised and contained numerous fenestrations of varying size. The possible function and developmental origin of these gaps is unclear; however, their role in promoting omental adhesion formation and in the successful use of omental graft material is discussed.
Assuntos
Células Epiteliais/ultraestrutura , Epitélio/ultraestrutura , Omento/ultraestrutura , Tecido Adiposo/fisiologia , Tecido Adiposo/ultraestrutura , Idoso , Idoso de 80 Anos ou mais , Animais , Vasos Sanguíneos/fisiologia , Vasos Sanguíneos/ultraestrutura , Células Epiteliais/fisiologia , Epitélio/fisiologia , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Omento/irrigação sanguínea , Omento/fisiologia , Peritonite/complicações , Peritonite/fisiopatologia , Especificidade da Espécie , Aderências Teciduais/fisiopatologiaRESUMO
PURPOSE: This study was designed to investigate the vascular structure and angiogenic activity of human peritoneal adhesions. METHODS: Adhesions were collected from patients undergoing laparotomy (n=32). Histologic features were documented and the distribution of mature and immature vascular markers were determined by immunolocalization and quantified by image analysis. The three-dimensional organization of blood vessels was investigated by confocal microscopy. Expression of vascular endothelial growth factor A, its receptor flk-1, and proliferating cell nuclear antigen were assessed by immunohistochemistry as indicators of angiogenic activity. RESULTS: Adhesions were found to be vascularized structures comprising bundles of collagen, interspersed with varying amounts of adipose tissue. Functional blood vessels expressed recognized vascular markers (vWF, CD34, alpha-SMA, and CD105) and formed a branching network similar to that of the peritoneum. Those adhesions expressing vascular endothelial growth factor A and its receptor showed significantly higher numbers of immature vessels as defined by expression of CD105. Omental adhesions (n=16) contained significantly more adipose tissue (P<0.05) and displayed a higher microvessel density (P<0.01) but lower cellularity (P<0.05) compared with nonomental adhesions (n=16). CONCLUSIONS: All adhesions contained functional blood vessels and most showed evidence of cell proliferation. The presence of vascular endothelial growth factor A and its receptor in human adhesions suggests ongoing angiogenic activity. This study demonstrates that adhesions are vascular structures with evidence of tissue remodeling and suggests potential for new prevention strategies involving antiangiogenic therapies.