Alveolar Microstructure Research Articles

Pulmonary functional MRI: Detecting the structure-function pathologies that drive asthma symptoms and quality of life.

Pulmonary functional MRI (PfMRI) using inhaled hyperpolarized, radiation-free gases (such as 3 He and 129 Xe) provides a way to directly visualize inhaled gas distribution and ventilation defects (or ventilation heterogeneity) in real time with high spatial (~mm3 ) resolution. Both gases enable quantitative measurement of terminal airway morphology, while 129 Xe uniquely enables imaging the transfer of inhaled gas across the alveolar-capillary tissue barrier to the red blood cells. In patients with asthma, PfMRI abnormalities have been shown to reflect airway smooth muscle dysfunction, airway inflammation and remodelling, luminal occlusions and airway pruning. The method is rapid (8-15 s), cost-effective (~$300/scan) and very well tolerated in patients, even in those who are very young or very ill, because unlike computed tomography (CT), positron emission tomography and single-photon emission CT, there is no ionizing radiation and the examination takes only a few seconds. However, PfMRI is not without limitations, which include the requirement of complex image analysis, specialized equipment and additional training and quality control. We provide an overview of the three main applications of hyperpolarized noble gas MRI in asthma research including: (1) inhaled gas distribution or ventilation imaging, (2) alveolar microstructure and finally (3) gas transfer into the alveolar-capillary tissue space and from the tissue barrier into red blood cells in the pulmonary microvasculature. We highlight the evidence that supports a deeper understanding of the mechanisms of asthma worsening over time and the pathologies responsible for symptoms and disease control. We conclude with a summary of approaches that have the potential for integration into clinical workflows and that may be used to guide personalized treatment planning.

Quantitative analysis of speckle-based X-ray dark-field imaging using numerical wave-optics simulations

The dark-field signal measures the small-angle scattering strength and provides complementary diagnostic information. This is of particular interest for lung imaging due to the pronounced small-angle scatter from the alveolar microstructure. However, most dark-field imaging techniques are relatively complex, dose-inefficient, and require sophisticated optics and highly coherent X-ray sources. Speckle-based imaging promises to overcome these limitations due to its simple and versatile setup, only requiring the addition of a random phase modulator to conventional X-ray equipment. We investigated quantitatively the influence of sample structure, setup geometry, and source energy on the dark-field signal in speckle-based X-ray imaging with wave-optics simulations for ensembles of micro-spheres. We show that the dark-field signal is accurately predicted via a model originally derived for grating interferometry when using the mean frequency of the speckle pattern power spectral density as the characteristic speckle size. The size directly reflects the correlation length of the diffuser surface and did not change with energy or propagation distance within the near-field. The dark-field signal had a distinct dependence on sample structure and setup geometry but was also affected by beam hardening-induced modifications of the visibility spectrum. This study quantitatively demonstrates the behavior of the dark-field signal in speckle-based X-ray imaging.

Lung microstructure in adolescent idiopathic scoliosis before and after posterior spinal fusion.

Adolescent idiopathic scoliosis (AIS) is associated with decreased respiratory quality of life and impaired diaphragm function. Recent hyperpolarized helium (HHe) MRI studies show alveolarization continues throughout adolescence, and mechanical forces are known to impact alveolarization. We therefore hypothesized that patients with AIS would have alterations in alveolar size, alveolar number, or alveolar septal dimensions compared to adolescents without AIS, and that posterior spinal fusion (PSF) might reverse these differences. We conducted a prospective observational trial using HHe MRI to test for changes in alveolar microstructure in control and AIS subjects at baseline and one year. After obtaining written informed consent from subjects' legal guardians and assent from the subjects, we performed HHe and proton MRI in 14 AIS and 16 control subjects aged 8-21 years. The mean age of control subjects (12.9 years) was significantly less than AIS (14.9 years, p = 0.003). At baseline, there were no significant differences in alveolar size, number, or alveolar duct morphometry between AIS and control subjects or between the concave (compressed) and convex (expanded) lungs of AIS subjects. At one year after PSF AIS subjects had an increase in alveolar density in the formerly convex lung (p = 0.05), likely reflecting a change in thoracic anatomy, but there were no other significant changes in lung microstructure. Modeling of alveolar size over time demonstrated similar rates of alveolar growth in control and AIS subjects in both right and left lungs pre- and post-PSF. Although this study suffered from poor age-matching, we found no evidence that AIS or PSF impacts lung microstructure. Trial registration: Clinical trial registration number NCT03539770.

Dry period heat stress induces microstructural changes in the lactating mammary gland.

The bovine dry period is a non-lactating period between consecutive lactations characterized by mammary gland involution and redevelopment phases to replace senescent mammary epithelial cells with active cells primed for the next lactation. Dairy cows exposed to heat stress during the dry period experience milk yield reductions between 3–7.5 kg/d in the next lactation, partially attributed to processes associated with mammary cell growth and turnover during the dry period. However, the carry-over impact of dry period heat stress on mammary morphology during lactation has yet to be determined. In the current study, we hypothesized that exposure to heat stress during the dry period would alter alveolar microstructure and cellular turnover (i.e. proliferation and apoptosis) during lactation. Cows were either subjected to heat stress (HT, access to shade; n = 12) or cooling (CL, access to shade, fans, and soakers; n = 12) for a 46 d dry period. Upon calving, all cows were treated similarly with access to cooling for their entire lactation. Six cows per treatment were randomly selected for mammary gland biopsies at 14, 42, and 84 days in milk. Tissues were sectioned and stained for histological analysis. During lactation, HT cows produced 4 kg less colostrum and 3.7 kg less milk compared with CL cows. Lactating mammary gland microstructure was impacted after exposure to dry period heat stress; HT cows had fewer alveoli and a higher proportion of connective tissue in the mammary gland relative to CL cows, however alveolar area was similar between treatments. Rates of mammary epithelial cell proliferation and apoptosis were similar between treatment groups. This suggests that heat stress exposure during the dry period leads to reductions in milk yield that could be caused, in part, by a reduction in alveoli number in the lactating mammary gland but not to dynamic alterations in cellular turnover once lactation is established.

The role of three-dimensionality and alveolar pressure in the distribution and amplification of alveolar stresses

Alveolar stresses are fundamental to enable the respiration process in mammalians and have recently gained increasing attention due to their mechanobiological role in the pathogenesis and development of respiratory diseases. Despite the fundamental physiological role of stresses in the alveolar wall, the determination of alveolar stresses remains challenging, and our current knowledge is largely drawn from 2D studies that idealize the alveolar septal wall as a spring or a planar continuum. Here we study the 3D stress distribution in alveolar walls of normal lungs by combining ex-vivo micro-computed tomography and 3D finite-element analysis. Our results show that alveolar walls are subject to a fully 3D state of stresses rather than to a pure axial stress state. To understand the contributions of the different components and deformation modes, we decompose the stress tensor field into hydrostatic and deviatoric components, which are associated with isotropic and distortional stresses, respectively. Stress concentrations arise in localized regions of the alveolar microstructure, with magnitudes that can be up to 27 times the applied alveolar pressure. Interestingly, we show that the stress amplification factor strongly depends on the level of alveolar pressure, i.e, stresses do not scale proportional to the applied alveolar pressure. In addition, we show that 2D techniques to assess alveolar stresses consistently overestimate the stress magnitude in alveolar walls, particularly for lungs under high transpulmonary pressure. These findings take particular relevance in the study of stress-induced remodeling of the emphysematous lung and in ventilator-induced lung injury, where the relation between transpulmonary pressure and alveolar wall stress is key to understand mechanotransduction processes in pneumocytes.

Contrast-to-noise ratios and thickness-normalized, ventilation-dependent signal levels in dark-field and conventional in vivo thorax radiographs of two pigs.

Lung tissue causes significant small-angle X-ray scattering, which can be visualized with grating-based X-ray dark-field imaging. Structural lung diseases alter alveolar microstructure, which often causes a dark-field signal decrease. The imaging method provides benefits for diagnosis of such diseases in small-animal models, and was successfully used on porcine and human lungs in a fringe-scanning setup. Micro- and macroscopic changes occur in the lung during breathing, but their individual effects on the dark-field signal are unknown. However, this information is important for quantitative medical evaluation of dark-field thorax radiographs. To estimate the effect of these changes on the dark-field signal during a clinical examination, we acquired in vivo dark-field chest radiographs of two pigs at three ventilation pressures. Pigs were used due to the high degree of similarity between porcine and human lungs. To analyze lung expansion separately, we acquired CT scans of both pigs at comparable posture and ventilation pressures. Segmentation, masking, and forward-projection of the CT datasets yielded maps of lung thickness and logarithmic lung attenuation signal in registration with the dark-field radiographs. Upon correlating this data, we discovered approximately linear relationships between the logarithmic dark-field signal and both projected quantities for all scans. Increasing ventilation pressure strongly decreased dark-field extinction coefficients, whereas the ratio of lung dark-field and attenuation signal changed only slightly. Furthermore, we investigated ratios of dark-field and attenuation noise levels at realistic signal levels via calculations and phantom measurements. Dark-field contrast-to-noise ratio (CNR) per lung height was 5 to 10% of the same quantity in attenuation. We conclude that better CNR performance in the dark-field modality is typically due to greater anatomical noise in the conventional radiograph. Given the high physiological similarity of human and porcine lungs, the presented thickness-normalized, ventilation-dependent values allow estimation of dark-field activity of human lungs of variable size and inspiration, which facilitates the design of suitable clinical imaging setups.

Translation from murine to human lung imaging using x-ray dark field radiography: A simulation study.

Recent studies on murine models have demonstrated the potential of dark field (DF) x-ray imaging for lung diseases. The alveolar microstructure causes small angle scattering, which is visualised in DF images. Whether DF imaging works for human lungs is not a priori guaranteed as human alveoli are larger and system settings for murine imaging will probably have to be adapted. This work examines the potential of translating DF imaging to human lungs. The DF contrast due to murine and human lung models was studied using numerical wave propagation simulations, where the lungs were modelled as a volume filled with spheres. Three sphere diameters were used: 39, 60 and 80 μm for the murine model and 200, 300 and 400 μm spheres for the human model. System settings applied for murine lung response modelling were taken from a prototype grating interferometry scanner used in murine lung experiments. The settings simulated for human lung imaging simulations combine the requirements for grating interferometry and conventional chest RX in terms of x-ray energy and pixel size. The DF signal in the simulated murine model was consistent with results from experimental DF data. The simulated linear diffusion coefficient for medium alveoli diameters was found to be (1.31±0.01)⋅10−11 mm-1, 120 times larger than those of human lung tissue ((1.09±0.01)⋅10−13 mm-1). However, as the human thorax is typically a factor 15 times larger than that of murine animals, the overall DF effect in human lungs remains substantial. At the largest lung thickness and for the DF setup simulated, human lungs have an estimated DF response of around 0.31 and murine lungs of 0.23. Dark field imaging can therefore be considered a promising modality for use in human lung imaging.

Antibacterial Micelles with Vancomycin-Mediated Targeting and pH/Lipase-Triggered Release of Antibiotics

Antibiotic delivery systems play an important role in increasing the efficacy while reducing the off-target toxicity and antibiotic resistance. Though bacterial infections share pathophysiological pathways similar to tumor tissues, few delivery systems have achieved bacterial targeting and on-demand release of antibiotics. In the current study, amphiphilic poly(ethylene glycol)-poly(ε-caprolactone) (PECL) copolymers are conjugated with vancomycin (VAN) as targeting ligands via pH-cleavable hydrazone bonds to obtain micelle carriers (Van-hyd-PECL). Subsequently, ciprofloxacin (CIP) is encapsulated to obtain Van-hyd-PECL/Cip micelles with an average size of 77 nm and a CIP loading amount of 4.5%. The poly(ethylene glycol) shells and the extension of VAN moieties on the micelle surface enhance the blood circulation and selective recognition of bacteria. The deshielding of VAN shells under acidic conditions disrupts the hydrophobic/hydrophilic balance leading to an increase in micelle sizes, which facilitates the degradation of poly(ε-caprolactone) by lipase overexpressed in the infection site and the release of encapsulated CIP for bacterial destruction. The micelle treatment has improved the survival of Pseudomonas aeruginosa-infected mice and reduced the bacterial burdens and alveolar injuries in lungs, compared with free drugs and micelles without inoculation of VAN moieties. Three doses of Van-hyd-PECL/Cip micelles further extend the animal survival, decrease the bacterial colonization in lungs, and almost restore the normal alveolar microstructure. In this regard, this study has demonstrated a strategy to enhance the bacterial targeting of micelles via an antibiotic (VAN) and to sequentially trigger the release of antibiotics (VAN and CIP) at the infection site.

Outside Back Cover: Biotechnology Journal 1/2018

Biotechnology JournalVolume 13, Issue 1 1870005 Outside Back CoverFree Access Outside Back Cover: Biotechnology Journal 1/2018 First published: 10 January 2018 https://doi.org/10.1002/biot.201870005AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Graphical Abstract Spectral confocal microscopy of living human lungs reveals the alveolar microstructure. Green calcein staining shows living cells constituting the alveolar septum, where the gas exchange takes place. The cells are attached to a tight and intertwined collagen and elastin network (grey) from which dying cells (red) detach into the air space (black areas). Image adjusted for illustration. This is reported by Katja Zscheppang, Johanna Berg, Sarah Hedtrich, Leonie Verheyen, Darcy E. Wagner, Norbert Suttorp, Stefan Hippenstiel and Andreas C. Hocke, article number 1700341. Volume13, Issue1Special Issue: Biotech Methods and AdvancesJanuary 20181870005 RelatedInformation

Finite element modeling of 129Xe diffusive gas exchange NMR in the human alveoli

Existing models of 129Xe diffusive exchange for lung microstructural modeling with time-resolved MR spectroscopy data have considered analytical solutions to one-dimensional, homogeneous models of the lungs with specific assumptions about the alveolar geometry. In order to establish a model system for simulating the effects of physiologically-realistic changes in physical and microstructural parameters on 129Xe exchange NMR, we have developed a 3D alveolar capillary model for finite element analysis. To account for the heterogeneity of the alveolar geometry across the lungs, we have derived realistic geometries for finite element analysis based on 2D histological samples and 3D micro-CT image volumes obtained from ex vivo biopsies of lung tissue from normal subjects and patients with interstitial lung disease. The 3D alveolar capillary model permits investigation of the impact of alveolar geometrical parameters and diffusion and perfusion coefficients on the in vivo measured 129Xe CSSR signal response. The heterogeneity of alveolar microstructure that is accounted for in image-based models resulted in considerable alterations to the shape of the 129Xe diffusive uptake curve when compared to 1D models. Our findings have important implications for the future design and optimization of 129Xe MR experiments and in the interpretation of lung microstructural changes from this data.

Lung tissue engineering and preservation of alveolar microstructure using a novel casting method

We used a rat model to decellularize and seed alveolar cells on a three-dimensional lung scaffold to preserve alveolar microarchitecture. We verified the preservation of terminal respiratory structure by casting and by scanning electron microscopy (SEM) of the casts after decellularization. Whole lungs were obtained from 12 healthy Sprague-Dawley rats, cannulated through the trachea under sterile conditions, and decellularized using a detergent-based method. Casting of both natural and decellularized lungs was performed to verify preservation of the inner microstructure of scaffolds for further cell seeding. Alveolar cell seeding was performed using green fluorescent protein (GFP) lung cells and non-GFP lung cells, and a peristaltic pump. We assessed cell seeding using histological and immunohistochemical staining, and enzymatic evaluation. All cellular components were removed completely from the scaffolds, and histological staining and SEM of casts were used to verify the preservation of tissue structure. Tensile tests verified conservation of biomechanical properties. The hydroxyproline content of decellularized lungs was similar to native lung. Histological and immunohistochemical evaluations showed effective cell seeding on decellularized matrices. Enzymatic measurement of trypsin and alpha 1 antitrypsin suggested the potential functional properties of the regenerated lungs. Casts produced by our method have satisfactory geometrical properties for further cell seeding of lung scaffolds. Preservation of micro-architecture and terminal alveoli that was confirmed by SEM of lung casts increases the probability of an effective cell seeding process.

Hyperpolarized 3He and 129Xe magnetic resonance imaging apparent diffusion coefficients: physiological relevance in older never- and ex-smokers.

Noble gas pulmonary magnetic resonance imaging (MRI) is transitioning away from 3He to 129Xe gas, but the physiological/clinical relevance of 129Xe apparent diffusion coefficient (ADC) parenchyma measurements is not well understood. Therefore, our objective was to generate 129Xe MRI ADC for comparison with 3He ADC and with well‐established measurements of alveolar structure and function in older never‐smokers and ex‐smokers with chronic obstructive pulmonary disease (COPD). In four never‐smokers and 10 COPD ex‐smokers, 3He (b = 1.6 sec/cm2) and 129Xe (b = 12, 20, and 30 sec/cm2) ADC, computed tomography (CT) density‐threshold measurements, and the diffusing capacity for carbon monoxide (DLCO) were measured. To understand regional differences, the anterior–posterior (APG) and superior–inferior (∆SI) ADC differences were evaluated. Compared to never‐smokers, COPD ex‐smokers showed greater 3He ADC (P = 0.006), 129Xe ADCb12 (P = 0.006), and ADCb20 (P = 0.006), but not for ADCb30 (P > 0.05). Never‐smokers and COPD ex‐smokers had significantly different APG for 3He ADC (P = 0.02), 129Xe ADCb12 (P = 0.006), and ADCb20 (P = 0.01), but not for ADCb30 (P > 0.05). ∆SI for never‐ and ex‐smokers was significantly different for 3He ADC (P = 0.046), but not for 129Xe ADC (P > 0.05). There were strong correlations for DLCO with 3He ADC and 129Xe ADCb12 (both r = −0.95, P < 0.05); in a multivariate model 129Xe ADCb12 was the only significant predictor of DLCO (P = 0.049). For COPD ex‐smokers, CT relative area <−950 HU (RA950) correlated with 3He ADC (r = 0.90, P = 0.008) and 129Xe ADCb12 (r = 0.85, P = 0.03). In conclusion, while 129Xe ADCb30 may be appropriate for evaluating subclinical or mild emphysema, in this small group of never‐smokers and ex‐smokers with moderate‐to‐severe emphysema, 129Xe ADCb12 provided a physiologically appropriate estimate of gas exchange abnormalities and alveolar microstructure.

Expansion Thoracoplasty Affects Lung Growth and Morphology in a Rabbit Model: A Pilot Study

BackgroundThoracic insufficiency syndrome represents a novel form of postnatal restrictive respiratory disease occurring in children with early-onset scoliosis and chest wall anomalies. Expansion thoracoplasty improves lung volumes in children with thoracic insufficiency syndrome; however, how it affects lung development is unknown.Questions/purposesUsing a rabbit model of thoracic insufficiency syndrome, we evaluated the effect of expansion thoracoplasty on the response of biologic mechanisms in the alveolar microstructure.MethodsUsing archived material from a previous experiment, 10 4-week-old New Zealand rabbits were divided into three groups: normal (n = 3), disease (n = 3), and treated (n = 4). Left ribs four to eight were tethered in seven rabbits at age 5 weeks to induce hypoplasia of the left hemithorax (disease). At age 10 weeks, four of these rabbits were treated by expansion thoracoplasty (treated). At age 24 weeks, lungs were excised and processed. Alveolar density and parenchymal airspace were measured on histologic sections. Immunohistochemistry was performed for vascular endothelial growth factor receptor 2 (angiogenesis), KI-67 (cell proliferation), and RAM-11 (macrophages).ResultsAlveolar walls were poorly perfused and airspace fraction was larger (emphysematous) in disease rabbits than normal or treated rabbits. Immunohistochemistry provided inconclusive evidence to support the concept that pulmonary hypoplasia is induced by thoracic insufficiency syndrome and controlled by expansion thoracoplasty.ConclusionsTreatment of thoracic insufficiency syndrome by expansion thoracoplasty may prevent emphysematous changes in the alveolar microstructure, thereby enhancing gas exchange.Clinical RelevanceCreating an animal model for thoracic insufficiency syndrome should provide insight into the effect of expansion thoracoplasty on lung development otherwise clinically unattainable.

Reflectance microscopy techniques for 3D imaging of the alveolar structure

Lung disease involving the alveoli and distal bronchioles are poorly understood and most commonly studied indirectly via lung function tests. Available imaging tools for the non-destructive assessment of the alveolar structure include X-ray computed tomography, intra-vital fluorescence microscopy and Optical Coherence Tomography, which are either limited by long acquisition time, inadequate resolution and contrast, or shallow imaging depth. In this study, we investigated the potential of two high-resolution reflectance microscopy imaging techniques, Spectrally Encoded Confocal Microscopy (SECM; 1µm (x) x 1µm (y) x 5µm (z) resolution) and Full Field Optical Coherence Microscopy (FFOCM; 1µm (x) x 1µm (y) x 1µm (z) resolution), for imaging alveolar microstructural detail. Two mouse lung samples were imaged with both SECM and FFOCM. The specimens were inflation-fixed using a modified Heitzman fixation technique at 20 cm H2O pressure. They were cut in 500mm thick slices and water immersed for imaging. Images were obtained and analyzed to determine whether or not the resolution and contrast of these techniques are sufficient to visualize the fine structures of the alveolar wall. Alveolar microstructure could be resolved in three dimensions in images obtained by both technologies. Alveolar septal walls from multiple layers could be clearly identified while sub-cellular structures such as nuclei were also visible in the SECM technique. In conclusion, we have demonstrated that two imaging technologies provide important sub-cellular detail that is required to study alveolar microstructure. Future research to develop these imaging modalities further so that they may be used in vivo is merited.

Evaluation of lung inflammation induced by intratracheal administration of LPS in mice: comparison between MRI and histology

To study the in vivo effect of intratracheal administration of lipopolysaccharide (LPS) in mice by magnetic resonance imaging (MRI) and to investigate the correlation with ex vivo histological evaluation of lung inflammation and oedema. LPS (or phosphate buffered saline) was administered intratracheally to thirty male Balb/C mice at a concentration of 0.3 mg/ml in a total volume of 100 microl. Animals were divided into fifteen LPS-treated and fifteen control mice. MR images were acquired 24 h after challenge in freely breathing animals with standard ECG-gated Gradient-Echo (GRE) sequences and, in a limited number of animals, with ECG-gated Ultrashort-echo time (UTE) sequences. After MRI, animals were sacrificed, and lungs were fixed and processed for histological analysis of the total volume of healthy lung tissue. GRE images revealed the presence of high intensity signal in lungs of LPS-treated mice that was attributable to oedema caused by alveolar inflammation. In histological slices, regions of alterations in the normal alveolar microstructure were observed that could account for MRI findings. A good correlation was observed between the volumes of lesioned tissue measured by MRI and by histology. The volume of the lesion detected by GRE sequences was lower than the volume detected by UTE sequences. The effect of intratracheal administration of LPS in mice was investigated by MRI and histology. A good correlation was observed between GRE-MRI and histological findings. MR images obtained with UTE sequences appear to be more sensitive to the presence of lesions than those obtained by standard GRE acquisitions.

Hyperpolarized Gas MR Imaging of the Lung: Current Status as a Research Tool

Hyperpolarized gas magnetic resonance imaging has been explored extensively as a promising tool for the quantitative evaluation of regional pulmonary pathophysiology. This noninvasive technique is capable of providing both structural information down to the level of the alveolar microstructure and functional information, such as dynamic ventilation, intrapulmonary partial pressure of oxygen, and alveolar surface area. This study reviews the role of hyperpolarized 3-helium and 129-xenon magnetic resonance imaging in this research.

MRI of the lungs using hyperpolarized noble gases.

The nuclear spin polarization of the noble gas isotopes (3)He and (129)Xe can be increased using optical pumping methods by four to five orders of magnitude. This extraordinary gain in polarization translates directly into a gain in signal strength for MRI. The new technology of hyperpolarized (HP) gas MRI holds enormous potential for enhancing sensitivity and contrast in pulmonary imaging. This review outlines the physics underlying the optical pumping process, imaging strategies coping with the nonequilibrium polarization, and effects of the alveolar microstructure on relaxation and diffusion of the noble gases. It presents recent progress in HP gas MRI and applications ranging from MR microscopy of airspaces to imaging pulmonary function in patients and suggests potential directions for future developments.

Healthy lungs tolerate repetitive collapse and reopening during short periods of mechanical ventilation.

The possible occurrence of lung damage if alveolar units are allowed to collapse and reopen breath by breath during mechanical ventilation with normal tidal volumes was investigated. Anaesthetised, paralysed, open chest rabbits were subjected to either intrathoracic negative (NEEP; n = 6) or positive (PEEP; n = 6) end-expiratory pressure during volume controlled mechanical ventilation. Both experimental settings were preceded by a 30 min control period and followed by a 30 min recovery period during which a PEEP of 0.2 kPa was maintained. Pao2 and pulmonary compliance deteriorated significantly in the NEEP group during the experimental period and compared to ventilation with PEEP. Partial restoration of lung mechanics and blood gases was achieved during the recovery period. After an alveolar recruitment manoeuvre, this recovery was complete. Lung clearance studied by depositing an aerosol of technetium-99m-labelled diethylenetriamine pentaacetate (99mTc-DTPA) in the alveoli, was significantly faster during ventilation with NEEP compared to the PEEP group (P = 0.0002) as well as the control period (P = 0.0029). It did not recover completely during the recovery period but remained significantly faster. Light microscopic histology was normal in both groups with no evidence of inflammation or epithelial disruption. We conclude that previously healthy rabbit lungs show only a transient disturbance of lung mechanics and blood gases with repetitive collapse and re-expansion. The integrity of the alveolar microstructure is preserved. The disturbance in the alveolo-capillary permeability persists and may indicate surfactant related alveolo-capillary barrier dysfunction.

Disturbance of alveolar lining layer: effects on alveolar microstructure

To further study the influence of altered surface tensions on alveolar micromechanics, we analyzed the structure-function relationships in excised rabbit lungs filled with or rinsed by a fluorocarbon (approximately 15 mN/m) or by hexadecane (approximately 25 mN/m). The lungs were fixed and dehydrated by vascular perfusion, and the tissue samples were analyzed by light, transmission, and scanning electron microscopy. We made three observations. 1) Pressure-volume (P-V) loops hexadecane-filled lungs are shifted to the left and coincide with those of saline-filled lungs, indicating near-zero interfacial tension. In accordance, the alveolar microstructure and surface area of hexadecane-filled lungs resemble those of saline-filled lungs. 2) The P-V loops of fluorocarbon-filled lungs are not shifted to the left but coincide with those of fluorocarbon-rinsed lungs. Under both conditions, the alveolar microstructure is qualitatively identical and the alveolar surface areas are markedly reduced compared with normal air-filled lungs. These findings show that fluorocarbon-filled or fluorocarbon-rinsed lungs are subjected to similar interfacial tensions at the alveolar level. 3) Hexadecane-rinsed lungs show a pear-shaped P-V curve and a complex surface texture of peripheral air spaces. These results, together with in vitro observations, suggest a metamorphic interplay between lung surfactant and hexadecane in lining the surface and determining the surface tension. Evidently, the effects of foreign liquids introduced into the lung on the structure-function relationship cannot accurately be predicted from their in vitro surface tensions. This fact should be considered in the development of artificial surfactants.

Amplitude-modulated Ventilation: Effects in Acute Lung Injury and Characterization of Mechanisms of Action

Amplitude modulated ventilation, administered as intermittent deep breaths to guinea pigs with experimental ALl, resulted in an increase in lung compliance which lasted many respiratory cycles. The response was greater in magnitude and longer lasting than that observed in control animals. Viscoelastic dynamic responses of parenchymal tissue strips and surfactant film preparations from damaged and control animals used in a microstructural model of alveolar stability failed to adequately predict the time responses observed in the intact lung. We are currently developing a more detailed dynamic model of alveolar microstructure which we hope will allow us to better understand how AMV may work, and how we can plan rationale strategies for improving alveolar stability and overall lung function in ALI without promoting damage in previously intact regions of the lung.