RESUMEN
The emission of cerium oxide nanoparticles (CeO2) from diesel engines, using cerium compounds as a catalyst to lower the diesel exhaust particles, is a health concern. We have previously shown that CeO2 induced pulmonary inflammation and lung fibrosis. The objective of the present study was to investigate the modification of fibroblast function and the role of epithelial-mesenchymal transition (EMT) in CeO2-induced fibrosis. Male Sprague-Dawley rats were exposed to CeO2 (0.15 to 7mg/kg) by a single intratracheal instillation and sacrificed at various times post-exposure. The results show that at 28days after CeO2 (3.5mg/kg) exposure, lung fibrosis was evidenced by increased soluble collagen in bronchoalveolar lavage fluid, elevated hydroxyproline content in lung tissues, and enhanced sirius red staining for collagen in the lung tissue. Lung fibroblasts and alveolar type II (ATII) cells isolated from CeO2-exposed rats at 28days post-exposure demonstrated decreasing proliferation rate when compare to the controls. CeO2 exposure was cytotoxic and altered cell function as demonstrated by fibroblast apoptosis and aggregation, and ATII cell hypertrophy and hyperplasia with increased surfactant. The presence of stress fibers, expressed as α-smooth muscle actin (SMA), in CeO2-exposed fibroblasts and ATII cells was significantly increased compared to the control. Immunohistofluorescence analysis demonstrated co-localization of TGF-ß or α-SMA with prosurfactant protein C (SPC)-stained ATII cells. These results demonstrate that CeO2 exposure affects fibroblast function and induces EMT in ATII cells that play a role in lung fibrosis. These findings suggest potential adverse health effects in response to CeO2 nanoparticle exposure.
Asunto(s)
Células Epiteliales Alveolares/efectos de los fármacos , Cerio/toxicidad , Transición Epitelial-Mesenquimal/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Pulmón/efectos de los fármacos , Nanopartículas del Metal/toxicidad , Fibrosis Pulmonar/inducido químicamente , Actinas/metabolismo , Células Epiteliales Alveolares/metabolismo , Células Epiteliales Alveolares/patología , Animales , Líquido del Lavado Bronquioalveolar/química , Forma de la Célula/efectos de los fármacos , Células Cultivadas , Colágeno/metabolismo , Fibroblastos/metabolismo , Fibroblastos/patología , Hidroxiprolina/metabolismo , Pulmón/metabolismo , Pulmón/patología , Masculino , Péptidos/metabolismo , Fibrosis Pulmonar/metabolismo , Fibrosis Pulmonar/patología , Ratas Sprague-Dawley , Medición de Riesgo , Fibras de Estrés/efectos de los fármacos , Fibras de Estrés/metabolismo , Fibras de Estrés/patología , Factores de Tiempo , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Our laboratory has previously demonstrated that application of an antimicrobial spray product containing titanium dioxide (TiO(2)) generates an aerosol of titanium dioxide in the breathing zone of the applicator. The present report describes the design of an automated spray system and the characterization of the aerosol delivered to a whole body inhalation chamber. This system produced stable airborne levels of TiO(2) particles with a median count size diameter of 110 nm. Rats were exposed to 314 mg/m(3) min (low dose), 826 mg/m(3) min (medium dose), and 3638 mg/m(3) min (high dose) of TiO(2) under the following conditions: 2.62 mg/m(3) for 2 h, 1.72 mg/m(3) 4 h/day for 2 days, and 3.79 mg/m(3) 4 h/day for 4 days, respectively. Pulmonary (breathing rate, specific airway resistance, inflammation, and lung damage) and cardiovascular (the responsiveness of the tail artery to constrictor or dilatory agents) endpoints were monitored 24 h post-exposure. No significant pulmonary or cardiovascular changes were noted at low and middle dose levels. However, the high dose caused significant increases in breathing rate, pulmonary inflammation, and lung cell injury. Results suggest that occasional consumer use of this antimicrobial spray product should not be a hazard. However, extended exposure of workers routinely applying this product to surfaces should be avoided. During application, care should be taken to minimize exposure by working under well ventilated conditions and by employing respiratory protection as needed. It would be prudent to avoid exposure to children or those with pre-existing respiratory disease.
Asunto(s)
Antiinfecciosos/toxicidad , Arterias/efectos de los fármacos , Pulmón/efectos de los fármacos , Nanopartículas del Metal/toxicidad , Titanio/toxicidad , Acetilcolina/farmacología , Administración por Inhalación , Aerosoles , Albúminas/metabolismo , Animales , Arterias/fisiología , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Recuento de Células , L-Lactato Deshidrogenasa/metabolismo , Pulmón/fisiología , Masculino , Neutrófilos/citología , Neutrófilos/efectos de los fármacos , Tamaño de la Partícula , Fenilefrina/farmacología , Ratas , Ratas Sprague-Dawley , Pruebas de Función Respiratoria , Cola (estructura animal) , Vasoconstrictores/farmacología , Vasodilatadores/farmacologíaRESUMEN
Nanomaterials are frontier technological products used in different manufactured goods. Because of their unique physicochemical, electrical, mechanical, and thermal properties, single-walled carbon nanotubes (SWCNT) are finding numerous applications in electronics, aerospace devices, computers, and chemical, polymer, and pharmaceutical industries. SWCNT are relatively recently discovered members of the carbon allotropes that are similar in structure to fullerenes and graphite. Previously, we (47) have reported that pharyngeal aspiration of purified SWCNT by C57BL/6 mice caused dose-dependent granulomatous pneumonia, oxidative stress, acute inflammatory/cytokine responses, fibrosis, and decrease in pulmonary function. To avoid potential artifactual effects due to instillation/agglomeration associated with SWCNT, we conducted inhalation exposures using stable and uniform SWCNT dispersions obtained by a newly developed aerosolization technique (2). The inhalation of nonpurified SWCNT (iron content of 17.7% by weight) at 5 mg/m(3), 5 h/day for 4 days was compared with pharyngeal aspiration of varying doses (5-20 microg per mouse) of the same SWCNT. The chain of pathological events in both exposure routes was realized through synergized interactions of early inflammatory response and oxidative stress culminating in the development of multifocal granulomatous pneumonia and interstitial fibrosis. SWCNT inhalation was more effective than aspiration in causing inflammatory response, oxidative stress, collagen deposition, and fibrosis as well as mutations of K-ras gene locus in the lung of C57BL/6 mice.
Asunto(s)
Administración por Inhalación , Inflamación/etiología , Pulmón/efectos de los fármacos , Mutagénesis , Nanotubos de Carbono/efectos adversos , Estrés Oxidativo/efectos de los fármacos , Trastornos Respiratorios/inducido químicamente , Aerosoles/administración & dosificación , Animales , Carbono/farmacología , Femenino , Fibrosis , Inflamación/patología , Pulmón/patología , Ratones , Ratones Endogámicos C57BL , FaringeRESUMEN
Nanoparticles have a fundamental dimension of <100 nm. However, on suspension in media, agglomerates of nanoparticles are the more common structure. This is particularly evident in prior intratracheal instillation or aspiration studies of single-walled carbon nanotubes (SWCNT), in which granulomatous lesions encased by epithelioid macrophages were produced by large agglomerates. In this study, we tested the hypothesis of whether exposure to more dispersed SWCNT structures would alter pulmonary distribution and response. A dispersed preparation of single-walled carbon nanotubes (DSWCNT) with a mean diameter of 0.69 microm was given by pharyngeal aspiration to C57BL/6 mice. Electron microscopy demonstrated a highly dispersed, interstitial distribution of DSWCNT deposits by 1 day postexposure. Deposits were generally <1 microm. Macrophage phagocytosis of DSWCNT was rarely observed at any time point. Lung responses were studied by lavage and morphometry at 1 h, 1 day, 7 day, and 1 mo after a single DSWCNT exposure of 10 microg/mouse. Lung sections and lavage cells demonstrated an early, transient neutrophilic and inflammatory phase that rapidly resolved and was similar to that observed with large agglomerates. No granulomatous lesions or epithelioid macrophages were detected. Morphometric measurement of Sirius red staining was used to assess the connective tissue response. The average thickness of connective tissue in alveolar regions was 0.10 +/- 0.02, 0.09 +/- 0.02, 0.10 +/- 0.01, 0.48 +/- 0.04, and 0.88 +/- 0.19 microm for PBS and 1-h, 1-day, 7-day, and 1-mo postexposure groups, respectively. The results demonstrate that dispersed SWCNT are rapidly incorporated into the alveolar interstitium and that they produce an increase in collagen deposition.
Asunto(s)
Carbono/farmacología , Carbono/farmacocinética , Inhalación/fisiología , Pulmón/fisiología , Nanotubos , Animales , Peso Corporal , Gases/metabolismo , Pulmón/anatomía & histología , Pulmón/efectos de los fármacos , Pulmón/ultraestructura , Ratones , Microscopía Confocal , Microscopía Electrónica de Rastreo , Microscopía Fluorescente , Modelos Animales , Neutrófilos/efectos de los fármacos , Neutrófilos/fisiología , Neutrófilos/ultraestructura , Tamaño de los Órganos , Alveolos Pulmonares/anatomía & histología , Propiedades de SuperficieRESUMEN
As the result of a high prevalence of fixed airways obstruction in workers at a microwave popcorn manufacturing plant, we examined the hypothesis that vapors of butter flavoring used in the manufacture of microwave popcorn and other foods can produce airway injury in rats. Rats were exposed to vapors liberated from heated butter flavoring. Rats were exposed for 6 h by inhalation and were necropsied 1 day after exposure. The exposure was found by GC-MS analysis to be a complex mixture of various organic gases with the major peaks consisting of diacetyl (2,3-butanedione), acetic acid, acetoin (3-hydroxy-2-butanone), butyric acid, acetoin dimers, 2-nonanone, and delta-alkyl lactones. Diacetyl was used as a marker of exposure concentration. In the lung, butter flavoring vapors containing 285-371 ppm diacetyl caused multifocal, necrotizing bronchitis, which was most consistently present in the mainstem bronchus. Alveoli were unaffected. Butter flavoring vapors containing 203-371 ppm diacetyl caused necrosuppurative rhinitis, which affected all four levels of the nose. Within the posterior two nasal levels (T3 and T4), necrosis and inflammation was principally localized to the nasopharyngeal duct. Control rats were unaffected. Therefore, concentrations of butter flavoring vapors that can occur during the manufacture of foods are associated with epithelial injury in the nasal passages and pulmonary airways of rats.
Asunto(s)
Bronquios/patología , Diacetil/toxicidad , Aromatizantes/toxicidad , Mucosa Nasal/patología , Animales , Bronquios/efectos de los fármacos , Bronquios/metabolismo , Líquido del Lavado Bronquioalveolar/citología , Recuento de Células , Histocitoquímica , Exposición por Inhalación , Masculino , Microscopía Electrónica , Líquido del Lavado Nasal/citología , Mucosa Nasal/efectos de los fármacos , Mucosa Nasal/metabolismo , Necrosis , Ratas , Ratas Sprague-Dawley , Organismos Libres de Patógenos EspecíficosRESUMEN
Emphysema is commonly defined as enlargement of airspaces distal to terminal bronchioles accompanied by destruction of alveolar walls, but without obvious fibrosis. Morphometric techniques were used to correlate changes in components of the alveolar septa surrounding enlarged airspaces in human emphysema with the mean linear intercept (Lm) of those airspaces. Alveolar and capillary surface density decreased with increased Lm, but the ratio of these surface densities to each other remained close to normal for mild to moderate increases in Lm. This suggests that the decreased gas exchange observed in emphysema is initiated by a total loss of septa and not by selective pathological changes of the microvasculature. Increases in septal wall thickness directly correlated with increases in Lm. For the mild to moderate emphysema lesions included in this study, an increase of 100% in Lm correlated with a 130% increase in the relative volume of the alveolar septal interstitium. Significant increases occurred in both elastin (0.14 to 0.56 microm(3)/microm(2) basement membrane [BM]) and collagen (0.49 to 1. 63 microm(3)/microm(2) BM). The increase in elastin and collagen raises the possibility of a remodeling process in the connective matrix in alveolar walls. Whether or not the new connective tissue represents a disordered, nonfunctional regional response needs to be determined.
Asunto(s)
Alveolos Pulmonares/patología , Enfisema Pulmonar/patología , Anciano , Capilares/patología , Colágeno/ultraestructura , Tejido Conectivo/patología , Tejido Conectivo/ultraestructura , Elastina/ultraestructura , Femenino , Humanos , Masculino , Persona de Mediana Edad , Alveolos Pulmonares/irrigación sanguínea , Alveolos Pulmonares/ultraestructuraRESUMEN
Nitric oxide (NO)*, the principal airborne pollutant generated from combustion processes such as gas stoves, tobacco smoke, and burning of fossil fuels, is being tested as a therapeutic agent in clinical trials. A prior morphometric study of rats exposed for 9 weeks to 0.5 parts per million (ppm) NO demonstrated focal degeneration of the alveolar interstitium and increased numbers of fenestrated alveolar septa (Mercer et al. 1995). The limited size and distribution of defects in this NO exposure did not alter alveolar surface area or other morphometric indicators of lung function, but were of interest as the responses to inhaled NO appeared to differ from those produced by other oxidants such as ozone (O3) and nitrogen dioxide (NO2). Nitric oxide exposures at the same concentration and duration as prior morphometric studies of O3 and NO2 were necessary in order to make a comparison. This was the purpose of the current study in which F344 rats were exposed for 6 weeks to air, 2 ppm NO, or 6 ppm NO. Following exposure, the lungs of NO- and air-exposed rats were preserved and prepared for electron microscopy. The lungs of replicate groups were lavaged and analyzed for protein content and antioxidants. Ultrastructural alterations due to exposure were determined by quantitative morphometric analyses and serial-section counts of the number of alveolar fenestrae. In contrast to the prior study of NO, there was no significant difference in the number of alveolar fenestrae/lung between control and NO-exposed groups. Morphometric analysis of the 6 ppm NO-exposure group demonstrated a significant increase from controls in the percentage of epithelial basement membrane covered by type II epithelial cells and a significant increase in the number of type II epithelial cells and airspace macrophages. At 2 ppm, only the percentage of epithelial basement membrane covered by type II epithelial cells was significant. No significant differences were found in lavage protein or in lavage ascorbic acid or glutathione content between clean-air controls and NO-exposed groups. Overall, the proinflammatory responses by type II epithelial cells and airspace macrophages following inhaled NO were comparable to those of O3 and NO2. These results, derived from experiments using significantly higher concentrations than in the prior study, demonstrate that inhaled NO produces a pattern of injury similar to that of other oxidants.
Asunto(s)
Contaminantes Atmosféricos/toxicidad , Óxido Nítrico/toxicidad , Alveolos Pulmonares/efectos de los fármacos , Administración por Inhalación , Animales , Líquido del Lavado Bronquioalveolar/inmunología , Relación Dosis-Respuesta a Droga , Epitelio/efectos de los fármacos , Masculino , Dióxido de Nitrógeno/toxicidad , Ozono/toxicidad , Agregación Plaquetaria/efectos de los fármacos , Alveolos Pulmonares/patología , Ratas , Ratas Endogámicas F344RESUMEN
In acute lung injury, a disturbed surfactant system may impair gas exchange. Previous evaluations of hyperoxia effects on surfactant proteins (SPs) followed exposures >1-2 days. To evaluate the effects of brief exposure to hyperoxia on the SP system, we exposed adult male rats to 95% O2 or air for 12, 36, and 60 h. SP-A, -B, and -C mRNAs were analyzed by Northern blot and semiquantitative in situ hybridization (ISH). SP-A and -B were analyzed in whole lung homogenates, lung lavage fluid, and fixed tissue by semiquantitative immunohistochemistry (IHC). All SP mRNAs were diminished at 12 h and rose to or exceeded control by 60 h as determined by Northern blot and ISH. These effects were seen mainly in the intensity of ISH signal per cell in both type II and bronchiolar epithelial (Clara) cells and to a lesser extent on numbers of positively labeled cells. SP-B declined to 50% of control in lavage at 12 h, but no changes in total lung SP-A and -B were seen. The number of SP-A positively labeled cells did not change, but SP-A label intensity measured by IHC in type II cells showed parallel results to Northern blots and ISH. The response of SP-A in Clara cells was similar. SP-B immunolabeling intensity rose in both type II and Clara cells throughout the exposure. SP-C ISH intensity fell at 12 h and was increased to two times control by 60 h of hyperoxia. Sharp declines in SP expression occurred by 12 h of 95% O2 and may affect local alveolar stability.
Asunto(s)
Bronquios/metabolismo , Oxígeno/farmacología , Proteolípidos/genética , Proteolípidos/metabolismo , Alveolos Pulmonares/metabolismo , Surfactantes Pulmonares/genética , Surfactantes Pulmonares/metabolismo , Animales , Northern Blotting , Western Blotting , Bronquios/efectos de los fármacos , Epitelio/efectos de los fármacos , Epitelio/metabolismo , Inmunohistoquímica , Hibridación in Situ , Masculino , Alveolos Pulmonares/efectos de los fármacos , Proteína A Asociada a Surfactante Pulmonar , Proteínas Asociadas a Surfactante Pulmonar , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Several cases of interstitial lung disease have been diagnosed among workers at a nylon flock plant, but the etiologic agent for the disease outbreak was unknown. The results of a medical survey and industrial hygiene study indicated that the dust present in the plant may be responsible. Thus, airborne dust collected at the plant was examined for its inflammatory potential in rat lungs. The endpoints measured were: (1) breathing rates, (2) differential cell counts of bronchoalveolar lavage cells, (3) alveolar macrophage (AM) chemiluminescence, (4) albumin concentration and matrix metalloprotease activities in the acellular fluid from the initial bronchoalveolar lavage, and (5) pulmonary histopathology. In the first study, rats received a single dose of the airborne dust sample (10 mg/kg body weight) by intratracheal (IT) instillation. At 1 d post-IT, all inflammatory endpoints were significantly increased versus controls, but by 29 d post-IT they did not differ significantly from controls. Histopathology demonstrated mild to moderate, multifocal, suppurative pneumonia, usually centered around bronchioles, at 1 d post-IT. At 29 d post-IT, pulmonary inflammation was minimal to mild and characterized by alveolar histocytosis usually restricted to the immediate area of retained bire-fringent fibers. In subsequent experiments, airborne dust was extracted with water and the dust (washed airborne dust) and water extract (soluble fraction) were separated by centrifugation for further study. Nylon tow dust was prepared in the laboratory by milling uncut nylon strands (called tow) that had not been treated with the finish or dyes that are commonly used in the flock plants. Rats were administered a single dose of a dust sample (10 mg/kg body weight) or the soluble fraction (1.3 ml/kg body weight) by IT administration and the same endpoints were measured at 1 d post-IT. The dust samples caused significant increases in all of the inflammatory endpoints; however, the soluble fraction was much less active. Histological analysis of the lungs 1 d post-IT confirmed lung inflammation was occurring and tended to center around bronchioles. The results suggest that: (1) nylon flocking generates particles of respirable size that can interact with AM in the lung and can be detected in the lung 29 d after exposure, (2) the dust samples examined cause an inflammatory response, (3) water-extractable agent(s) from airborne dust contribute only minimally to the inflammatory response, and (4) the acute inflammatory response to these dusts is substantial when compared to other pathologic occupational dusts previously examined in our laboratory.
Asunto(s)
Contaminantes Ocupacionales del Aire/toxicidad , Enfermedades Pulmonares Intersticiales/inducido químicamente , Nylons/toxicidad , Industria Textil , Enfermedad Aguda , Contaminantes Ocupacionales del Aire/análisis , Animales , Líquido del Lavado Bronquioalveolar/citología , Endotoxinas/análisis , Endotoxinas/toxicidad , Mediciones Luminiscentes , Pulmón/metabolismo , Pulmón/patología , Macrófagos Alveolares/efectos de los fármacos , Masculino , Metaloendopeptidasas/metabolismo , Nylons/análisis , Ratas , Ratas Sprague-Dawley , Albúmina Sérica/metabolismoRESUMEN
Surfactant protein D (SP-D), which has structural homology to C-type lectin binding regions, may play a role in host defense and has no known surfactant function. Because other surfactant proteins have been shown to be increased after prolonged periods of hyperoxia, we sought to evaluate the early effects of hyperoxia (95% O2) on expression of SP-D in the adult male rat lung. Animals were exposed to air or to 12, 36, or 60 h of 95% O2. Northern blot analysis of total lung RNA revealed marked SP-D mRNA increases at 12 h 95% O2 compared with air-exposed controls, with decreasing expression to near that of air-exposed animals by 60 h. Semiquantitative in situ RNA hybridization demonstrated parallel results, with increased numbers of labeled alveolar epithelial (AE) and bronchiolar epithelial (BE) cells at 12 h and increased intensity of labeled alveolar cells, compared with air-exposed controls. After 60 h of exposure to 95% O2, mRNA label intensity in AE and BE was decreased to levels near those seen in air-exposed animals. In contrast, Western blotting showed a decline in total lung SP-D with 95% O2 exposure, beginning at 12 h and continuing at 36 and 60 h, respectively. Semiquantitative immunohistochemistry demonstrated a decline in AE labeling parallel to the total lung Western blot results, but labeled total BE cell numbers increased (P = 0.10). Hyperoxia had differential effects on SP-D abundance in AE and BE cells, and therefore may influence the availability of SP-D to bind microbial pathogens in the airways depending on cell type and location.
Asunto(s)
Bronquios/metabolismo , Glicoproteínas/metabolismo , Oxígeno/administración & dosificación , Alveolos Pulmonares/metabolismo , Surfactantes Pulmonares/metabolismo , ARN Mensajero/metabolismo , Animales , Northern Blotting , Western Blotting , Epitelio/metabolismo , Glicoproteínas/genética , Hibridación in Situ , Masculino , Proteína D Asociada a Surfactante Pulmonar , Surfactantes Pulmonares/genética , ARN Mensajero/genética , Ratas , Ratas Sprague-DawleyRESUMEN
The initial pulmonary vasodilation that occurs during nitric oxide (. NO) inhalation does not appear to be maintained chronically in many cases. . NO may acutely relax vascular smooth muscle by increasing levels of guanosine 3',5'-cyclic monophosphate (cGMP), tumor necrosis factor (TNF)-alpha, and inducible nitric oxide synthase (iNOS) while decreasing levels of lipid peroxidation. It was hypothesized that the acute . NO-induced changes in cGMP, TNF-alpha, iNOS, and lipid peroxidation, all of which may mediate vasodilation, are transient rather than sustained. Lungs from rats kept in chambers containing 6 parts/million . NO for 1 h, 1 day, or 1 wk were analyzed for levels of . NO-induced vasodilatory mediators. Pulmonary cGMP, iNOS mRNA, and TNF-alpha were increased 1 h after . NO exposure but decreased to control values at later times. Levels of malonyl dialdehyde, an indicator of lipid peroxidation, were decreased at all times during . NO inhalation. As a whole, the data suggest that in lungs the vasodilatory mediators cGMP, iNOS, and TNF-alpha are only acutely and transiently elevated during inhalation of . NO, consistent with the initially positive clinical response to inhaled . NO that deteriorates over time.
Asunto(s)
GMP Cíclico/metabolismo , Pulmón/fisiología , Óxido Nítrico Sintasa/genética , Óxido Nítrico/farmacología , Transcripción Genética/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Adenilil Ciclasas/biosíntesis , Adenilil Ciclasas/genética , Administración por Inhalación , Animales , Aorta Torácica/citología , Aorta Torácica/enzimología , Células Cultivadas , Cartilla de ADN , Humanos , Interferón gamma/biosíntesis , Cinética , Peroxidación de Lípido/efectos de los fármacos , Pulmón/efectos de los fármacos , Masculino , Malondialdehído/análisis , Modelos Biológicos , Músculo Liso Vascular/citología , Músculo Liso Vascular/enzimología , Óxido Nítrico/administración & dosificación , Óxido Nítrico Sintasa/biosíntesis , Óxido Nítrico Sintasa de Tipo II , Reacción en Cadena de la Polimerasa , ARN Mensajero/biosíntesis , Ratas , Ratas Endogámicas F344 , Factores de Tiempo , Factor de Necrosis Tumoral alfa/biosíntesis , VasodilataciónRESUMEN
The cellular and tissue changes in the lungs of rats were studied using electron microscopy following 20 months exposure to a range of ozone levels from 0.12 to 1.0 ppm. Male and female Fischer 344 rats were exposed and morphometric methods were used to determine the volume, surface area, and cellular changes observed in bronchiole-alveolar duct regions following chronic ozone exposure. No major gender-related effects were observed in response to chronic inhalation of ozone nor were significant effects of ozone exposure found in either terminal bronchioles or the proximal alveolar regions in animals chronically exposed to 0.12 ppm ozone. The proximal alveolar regions of animals exposed for 20 months to 0.5 and 1.0 ppm ozone were significantly altered with exposure. The high-dose, long-term exposure to ozone resulted in a pronounced increase in volume of both the interstitium and epithelium in the proximal alveolar regions. The thickening of the epithelium was due to a change in tissue type from the normal squamous epithelium to a cuboidal epithelium similar, but not identical, to that found in terminal bronchioles. This bronchiolar epithelial metaplasia of proximal alveolar ducts, which was dose related, was composed of differentiated ciliated and Clara cells similar to those found in terminal bronchioles. In addition, unique cells which contained morphologic features of many different cell types were observed. These cells, which may represent stem cells or differentiated but transformed cells, were found associated with the bronchiolar metaplasia of alveolar ducts. In conjunction with the epithelial changes, cellular and matrix components in the interstitium were increased with chronic exposure to 0.5 and 1.0 ppm ozone. All matrix components were increased including collagen, elastin, and basement membrane, as well as other acellular spaces which did not contain identifiable structures. The total volume of interstitial fibroblasts was also increased in the high-dose exposure group. Alveolar macrophages were increased only in the 1.0 ppm exposed animals. The cell and tissue changes in the terminal bronchioles were less pronounced indicating a relative resistance of this tissue to ozone and mainly consisted of a change in cell type from ciliated to Clara cells in the 1.0 ppm exposed animals. The relative resistance of bronchiolar tissue to high concentration ozone exposure and the extensive bronchiolar epithelial metaplasia may be an adaptive mechanism following chronic ozone exposure.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Bronquios/efectos de los fármacos , Oxidantes Fotoquímicos/toxicidad , Ozono/toxicidad , Fibrosis Pulmonar/inducido químicamente , Animales , Bronquios/patología , Bronquios/ultraestructura , Femenino , Masculino , Metaplasia , Alveolos Pulmonares/efectos de los fármacos , Alveolos Pulmonares/patología , Alveolos Pulmonares/ultraestructura , Fibrosis Pulmonar/patología , Ratas , Ratas Endogámicas F344 , Factores SexualesRESUMEN
BACKGROUND: Nitric oxide (NO) and nitrogen dioxide (NO2) are common copollutants resulting from combustion processes such as the burning of fossil fuels and tobacco smoke. The relative toxicity of these two pollutants has not been adequately addressed. Separate low level exposures to each of these two pollutants were carried out to allow comparisons of relative health risks. EXPERIMENTAL DESIGN: Male rats were exposed to either NO or NO2 for 9 weeks at 0.5 ppm with twice daily, 1-hour spikes to 1.5 ppm. Lungs from five rats in each exposed group and from rats from a clean air control group were preserved by vascular perfusion of fixative and were embedded for sectioning. The number of fenestrations in alveolar septa of the lung was determined by using serial sections to directly count the number of fenestrae in a known volume of lung. RESULTS: The average number of fenestrae was 328 +/- 156 x 10(3) (mean +/- SE, n = 5) per lung in the NO group. In the NO2 exposure group, there were 99 +/- 42 x 10(3) fenestrae per lung. The number of fenestrae per lung in the controls (9 +/- 9 x 10(3)) was not statistically different from zero. The number of fenestrae in the NO group was significantly greater than that in the control or NO2 groups. Analyses of total parenchymal cells per lung demonstrated a statistically significant 29% reduction in the number of interstitial cells in the NO group. There were no significant differences in the numbers of other types of cells between the control and exposed groups. The thickness of the interstitial space was reduced in the NO group (0.24 +/- 0.02 microns versus 0.32 +/- 0.02 microns in controls) but not in the NO2 group (0.29 +/- 0.02 microns). Epithelial cell thickness did not differ between groups. CONCLUSIONS: Focal degeneration of interstitial cells, interstitial matrix, and connective tissue fibers is the principal injury resulting from low level NO exposure. NO is significantly more potent than NO2 in the production of these defects in the interstitial spaces of alveolar septa. Although limited in number and size, the formation of fenestrae by atrophy of the interstitial spaces is similar to the initial steps in an emphysema-like destruction of alveolar septa.
Asunto(s)
Óxido Nítrico/toxicidad , Dióxido de Nitrógeno/toxicidad , Alveolos Pulmonares/efectos de los fármacos , Animales , Masculino , Alveolos Pulmonares/patología , Ratas , Ratas Endogámicas F344RESUMEN
Morphometric techniques were used to examine cellular and tissue changes occurring in male and female rat lungs exposed to ozone for a prolonged time. F344/N rats were exposed to 0.0, 0.12, 0.5, or 1.0 parts per million (ppm)* ozone for six hours per day, five days per week, for 20 months. Changes in cell volume, cell surface ratios, and cellular characteristics were studied in the terminal bronchioles and in the proximal alveolar regions of the lungs. Animals exposed for 20 months to 0.5 or 1.0 ppm ozone demonstrated dramatic increases in the volume of interstitium and epithelium along the alveolar ducts. The thickening of the epithelium was caused by an epithelial metaplasia in which the normal squamous epithelium was modified to a cuboidal epithelium similar, but not identical, to the type found in terminal bronchioles. This bronchiolar epithelial metaplasia was directly related to dose of ozone, and was characterized by differentiated ciliated cells and Clara cells similar to those found in terminal bronchioles; undifferentiated cuboidal cells also were found in the animals exposed to 0.5 and 1.0 ppm ozone. A mild fibrotic response was seen in the animals exposed to 1.0 ppm ozone, with increases in both the interstitial matrix and cellular interstitium. The individual components of the interstitial matrix, including collagen, elastin, basement membrane, and acellular spaces, all were increased. The increase in cellular interstitium was due to an increase in the volume of interstitial fibroblasts. A slight inflammatory response, identified by an increase in alveolar macrophages, was observed in the animals exposed to 1.0 ppm. The terminal bronchioles were less affected than the proximal alveolar region by the ozone exposures, which may indicate a resistance of this tissue to ozone damage. The changes in the terminal bronchioles mainly consisted of a shift in cell type from ciliated to Clara cells in the animals exposed to 1.0 ppm ozone. The bronchiolar epithelial metaplasia observed in the proximal alveolar ducts may indicate that a protective mechanism develops in response to prolonged exposure to high concentrations of ozone.
Asunto(s)
Ozono/toxicidad , Alveolos Pulmonares/efectos de los fármacos , Alveolos Pulmonares/patología , Análisis de Varianza , Animales , Bronquios/efectos de los fármacos , Bronquios/patología , Relación Dosis-Respuesta a Droga , Epitelio , Femenino , Enfermedades Pulmonares Intersticiales/inducido químicamente , Enfermedades Pulmonares Intersticiales/patología , Macrófagos Alveolares/efectos de los fármacos , Macrófagos Alveolares/patología , Masculino , Metaplasia , Fibrosis Pulmonar/inducido químicamente , Fibrosis Pulmonar/patología , Ratas , Ratas Endogámicas F344 , Factores Sexuales , Factores de TiempoRESUMEN
Because the retractive forces due to surface tension decrease with increasing radius of curvature, there should be a greater contribution to lung recoil attributable to the stress-bearing role of elastic elements in the lung parenchyma of species with larger alveoli. To examine alterations in lung structure that may relate to this stress-bearing role, the lungs of mice, hamsters, rats, rabbits, rhesus monkeys, baboons, and humans were preserved by vascular perfusion of fixative. The number of alveoli per lung, alveolar radius of curvature, surface area, and volume were measured by serial section reconstruction. Electron-microscopic determinations were made of the volume fraction and thickness of the epithelium, interstitium, and endothelium and of the connective tissue fibers of the alveolar septa and the portions of alveolar septa that form the alveolar ducts. The thickness of the alveolar septal interstitium increased linearly with the increase in radius of curvature of alveoli. The increase in interstitial thickness in lungs with larger alveoli was paralleled by large increases in the volume of collagen and elastin fibers present in this space. Comparable changes in the thickness of connective tissue fibers in alveolar duct walls were also found. This study demonstrates species-related changes in the structure of alveolar septa and in lung collagen and elastin fibers that are consistent with connective tissue fibers having a greater stress-bearing role in both the alveolar septa and alveolar ducts of species with larger alveoli.
Asunto(s)
Alveolos Pulmonares/anatomía & histología , Animales , Colágeno/metabolismo , Tejido Conectivo/anatomía & histología , Tejido Conectivo/metabolismo , Tejido Conectivo/fisiología , Cricetinae , Elasticidad , Elastina/metabolismo , Humanos , Pulmón/anatomía & histología , Macaca mulatta , Ratones , Microscopía Electrónica de Rastreo , Papio , Alveolos Pulmonares/metabolismo , Alveolos Pulmonares/fisiología , Conejos , Ratas , Mecánica Respiratoria/fisiología , Especificidad de la Especie , Fijación del TejidoRESUMEN
Morphometric procedures were used to determine the number of cells, cell volume, cell diameter, and surface areas of the airways in human and rat lungs. Nuclear sizes of epithelial cells from human bronchi were significantly larger than other lung cell nuclei. The average volume of human ciliated cell nuclei was 310 +/- 30 microns 3 and 167 +/- 12 microns 3 in bronchi and bronchioles, respectively. The smaller nuclei of human bronchioles were comparable to those of alveolar cells. In the pseudostratified epithelium of human bronchi, basal cells had a large surface area in contact with the basement membrane (51.3 +/- 4.6 microns 2 per cell) when compared with ciliated (1.1 +/- 0.1 microns 2), goblet (7.6 +/- 1.2 microns 2), or other secretory cells (12.0 +/- 2.1 microns 2). In the first four airway generations distal to the trachea, basal cells account for 30% of the cells in human airway epithelium and 2% of the cells in rat airway epithelium. Total airway surface area from trachea to bronchioles was 2,471 +/- 320 and 27.2 +/- 1.7 cm2 in human and rat lungs, respectively. These direct measurements of airway surface area are less than half of the estimates based on current lung models. The total number of airway epithelial cells were 10.5 x 10(9) for human and 0.05 x 10(9) for rat lungs. For both species, there were 18 times more alveolar cells than bronchial epithelial cells.
Asunto(s)
Bronquios/citología , Pulmón/citología , Adulto , Animales , Membrana Basal/citología , Bronquios/ultraestructura , Núcleo Celular , Tamaño de la Célula , Células Epiteliales , Femenino , Humanos , Pulmón/ultraestructura , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Aerosolized fluorescent microspheres were used to study particle deposition in site-specific regions of the lung with confocal laser scanning microscopy. A nebulizer was used to aerosolize microspheres followed by passage through a heated discharging column to reduce static charge and to remove water surrounding each microsphere. Precoating of microspheres with albumin helped to minimize displacement during vascular fixation of the lungs. Confocal laser microscopy facilitated visualization of microspheres throughout the bronchial tree, ducts, and alveoli of the lungs. The use of fluorescent microspheres and confocal laser imaging provided distinct advantages compared with other methods to study lung particle deposition due to (1) the generation of single microspheres of uniform size by nebulization, (2) easy detection of microspheres in large slabs of microdissected lung tissues, (3) excellent resolution of tissue surfaces and microspheres for an infinite number of orientations and planes of section, and (4) the ability to visualize microspheres below fluid lining layers and on surfaces that could not easily be done by other methods of microscopy.
Asunto(s)
Aerosoles , Colorantes Fluorescentes , Pulmón/anatomía & histología , Microesferas , Animales , Masculino , Microscopía/métodos , Fagocitosis , Ratas , Ratas Sprague-DawleyRESUMEN
The distribution of ozone-induced injury across ventilatory units of the lungs was determined and compared with the predicted distribution of ozone dose across the same units to evaluate dose-response relationships. Sprague-Dawley rats were exposed to either 0.98 ppm ozone 8 h/day for 90 days or to filtered air only. En bloc microdissection was used to identify and isolate in longitudinal profile the bronchiole-alveolar duct junction, first pair of alveolar duct generations, and intervening bifurcation ridge. The first alveolar outpocketing along the bronchiolar wall of each isolation was used to identify the center of a series of concentric arcs radiating outward at 100-microns intervals across each ventilatory unit. The intercept lengths of each arc with the tissue of alveolar septal tips (edges) and alveolar walls were measured and expressed as a function of distance into the ventilatory unit. Relative ozone dose across the ventilatory unit was estimated using the geometry of the tracheobronchial tree and the volume and surface area distribution within individual ventilatory units. This mathematical model of ozone dose demonstrated a high degree of correlation to this measured tissue injury response. The findings of this study demonstrate that microdosimetry and microtoxicology can be used to determine dose-response relationships within the ventilatory unit and to assess questions of tissue sensitivity in ozone-induced lung injury.
Asunto(s)
Ozono/toxicidad , Sistema Respiratorio/efectos de los fármacos , Sistema Respiratorio/lesiones , Animales , Bronquios/efectos de los fármacos , Bronquios/lesiones , Bronquios/patología , Relación Dosis-Respuesta a Droga , Masculino , Microscopía Electrónica de Rastreo , Ozono/administración & dosificación , Alveolos Pulmonares/efectos de los fármacos , Alveolos Pulmonares/lesiones , Alveolos Pulmonares/patología , Ratas , Ratas Sprague-Dawley , Sistema Respiratorio/patologíaRESUMEN
Although total cell number has been determined for the alveolar region of the lungs of many species, it has not been calculated for the nonalveolar lung tissues. The oriented structure of airways and vessels makes the numerical assessment of cells in nonalveolar tissues difficult. This has led many investigators to use the number of cells in the alveolar region as a direct estimate of total lung cell number. To determine the number of cells in the nonalveolar lung tissues, the lungs of eight rats weighing 230 to 380 g were inflation-fixed and embedded in araldite, and 1.5-microns serial sections of the entire left lobe were cut and stained with methylene blue for light microscopy. The sections were then uniformly point-counted using computer-controlled distances between the fields to determine the fraction of points falling on air, blood, cellular tissue, and noncellular tissue for both the alveolar and the nonalveolar regions. The total volume of cell nuclei in each compartment was determined, and the total number of cells was calculated by dividing the total nuclear volume by the mean cell nuclear volume. It was found that 87% of the lung volume was alveolar, of which 6% was tissue and contained 725 x 10(6) cells. The nonalveolar region constituted 13% of the lung volume, of which 23% was tissue and contained 250 x 10(6) cells. The average rat lung therefore contains 975,000,000 cells, of which 74% was in alveolar tissues and 26% in nonalveolar tissues. On the basis of assays of isolated lung cells, there is an average of 7 pg DNA/cell.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Pulmón/citología , Alveolos Pulmonares/citología , Animales , Recuento de Células , Núcleo Celular , ADN/análisis , Estudios de Evaluación como Asunto , Pulmón/anatomía & histología , Análisis Numérico Asistido por Computador , Tamaño de los Órganos , Alveolos Pulmonares/anatomía & histología , Ratas , Ratas Sprague-DawleyRESUMEN
Ultrastructural changes in lung parenchymal elastic fibers were studied morphometrically 1, 4, and 12 wk after a single 12-unit dose of pancreatic elastase and in a saline-instilled control group. The mean linear intercept of the parenchymal air spaces was increased in the 1-, 4-, and 12-wk post-elastase instillation groups compared with age-matched controls. The volume of alveolar connective tissue fibers predominantly composed of elastin (elastic fibers) was decreased by 35% 1 wk after the instillation of elastase but returned to control levels by 4 wk. Although the total volume of elastic fibers was normal 12 wk after instillation of elastase, the volume of elastic fibers in alveolar entrance rings remained significantly reduced. In serial sections of elastic fibers, numerous gaps or separations in the normally continuous band of elastic fibers that encircle each alveolus were identified 1 wk after elastase instillation. There were 169 +/- 8 (SE), 62 +/- 32, and 12 +/- 6 gaps per millimeter of alveolar entrance ring circumference at 1, 4, and 12 wk, respectively, in the elastase-treated groups. The number of gaps at 12 wk was equivalent to two gaps or discontinuities in the elastic fibers of every alveolar entrance ring. No gaps or separations in elastic fibers were detected at 1, 4, or 12 wk in the control groups. These defects occur in concordance with the progression of air space enlargement and presumably contribute to the progression of air space enlargement that occurs after the elastin content of the tissue has returned to normal.