Howard W. Mitchell

Airway smooth muscle dynamics: a common pathway of airway obstruction in asthma

Excessive airway obstruction is the cause of symptoms and abnormal lung function in asthma. As airway smooth muscle (ASM) is the effecter controlling airway calibre, it is suspected that dysfunction of ASM contributes to the pathophysiology of asthma. However, the precise role of ASM in the series of events leading to asthmatic symptoms is not clear. It is not certain whether, in asthma, there is a change in the intrinsic properties of ASM, a change in the structure and mechanical properties of the noncontractile components of the airway wall, or a change in the interdependence of the airway wall with the surrounding lung parenchyma. All these potential changes could result from acute or chronic airway inflammation and associated tissue repair and remodelling. Anti-inflammatory therapy, however, does not “cure” asthma, and airway hyperresponsiveness can persist in asthmatics, even in the absence of airway inflammation. This is perhaps because the therapy does not directly address a fundamental abnormality of asthma, that of exaggerated airway narrowing due to excessive shortening of ASM. In the present study, a central role for airway smooth muscle in the pathogenesis of airway hyperresponsiveness in asthma is explored.

THE CAT LUNG STRIP AS AN in vitro PREPARATION OF PERIPHERAL AIRWAYS: A COMPARISON OF β‐ADRENOCEPTOR AGONISTS, AUTACOIDS AND ANAPHYLACTIC CHALLENGE ON THE LUNG STRIP AND TRACHEA

1 A new in vitro preparation, the isolated lung strip of the cat, is described for investigating the direct effect of drugs on the smooth muscle of the peripheral airways of the lung. The preparation comprises a thin strip of lung parenchyma which can be mounted in a conventional organ bath for isometric tension recording. Its pharmacological responses have been characterized and compared with the isolated tracheal preparation of the cat. 2 The lung strip exhibited an intrinsic tone which was relaxed by catecholamines, aminophylline and flufenamate. It was contracted strongly by histamine, prostaglandin F2α, acetylcholine, compound 48/80, potassium depolarizing solution and alternating current field stimulation. In contrast, the cat trachea was unresponsive to histamine and prostaglandin F2α and did not exhibit an intrinsic tone. 3 (–)‐Isoprenaline and (–)‐adrenaline were much more potent in relaxing the lung strip than the trachea. The potency order of relaxation responses to isoprenaline, adrenaline and (±)‐noradrenaline in the lung strip was isoprenaline > adrenaline > noradrenaline but in the trachea was isoprenaline > noradrenaline > adrenaline. 4 β2‐Adrenoceptor selective agonists salbutamol and terbutaline were more potent in the lung strip than the trachea, suggesting β2‐adrenoceptors predominated in the lung strip. Propranolol was equipotent in inhibiting isoprenaline relexations of the lung strip and trachea, whereas practolol was much less effective in inhibiting lung strip than trachea, further supporting a predominance of β2‐adrenoceptors in lung strip and β‐adrenoceptors in trachea. 5 Strong Schultz‐Dale type contractions were elicited in both lung strips and trachea by Ascaris lumbricoides antigen in actively sensitized cats. The initial phase of the contractile response of the lung strip following challenge was shown to be due to histamine release and was absent in the trachea. The delayed phase of the contraction which took several minutes to develop in both the mepyramine‐treated lung strip and trachea was not due to prostaglandins E1, F2α or bradykinin, the probable mediator being slow reacting substance of anaphylaxis (SRS‐A). 6 It is concluded that the isolated lung strip of the cat is useful as an in vitro model for investigating the effect of drugs on the smooth muscle of the peripheral airways of the lungs.

Measuring airway dimensions during bronchoscopy using anatomical optical coherence tomography

Airway dimensions are difficult to quantify bronchoscopically because of optical distortion and a limited ability to gauge depth. Anatomical optical coherence tomography (aOCT), a novel imaging technique, may overcome these limitations. This study evaluated the accuracy of aOCT against existing techniques in phantom, excised pig and in vivo human airways. Three comparative studies were performed: 1) micrometer-derived area measurements in 10 plastic tubes were compared with aOCT-derived area; 2) aOCT-derived airway compliance curves from excised pig airways were compared with curves derived using an endoscopic technique; and 3) airway dimensions from the trachea to subsegmental bronchi were measured using aOCT in four anaesthetised patients during bronchoscopy and compared with computed tomography (CT) measurements. Measurements in plastic tubes revealed aOCT to be accurate and reliable. In pig airways, aOCT-derived compliance measurements compared closely with endoscopic data. In human airways, dimensions measured with aOCT and CT correlated closely. Bland–Altman plots showed that aOCT diameter and area measurements were higher than CT measurements by 7.6% and 15.1%, respectively. Airway measurements using aOCT are accurate, reliable and compare favourably with existing imaging techniques. Using aOCT with conventional bronchoscopy allows real-time measurement of airway dimensions and could be useful clinically in settings where knowledge of airway calibre is required.

Modulation by the epithelium of the extent of bronchial narrowing produced by substances perfused through the lumen.

1 Airway narrowing was determined in vitro as a measure of bronchial reactivity. A bronchial segment from pig lung was perfused with a Krebs solution and the change in flow rate to drugs and small ions perfused intraluminally was compared with that obtained by application to the serosal surface. 2 The sensitivity (EC50) to acetylchloline was 30 times greater on the serosal surface than on the luminal surface. Concentrations of histamine and carbachol which had threshold responses on flow rate when perfused intraluminally virtually stopped flow on the serosal surface. Potassium depolarizing solutions (containing either KCl or K2SO4) and vanadate (VO−3) had little or no effect intraluminally but completely stopped flow through the bronchial segment when applied to the serosal surface, i.e. they closed off the airway. 3 After removal of the epithelium the sensitivity to drugs and K+ perfused intraluminally was increased to equal that on the serosal surface. 4 No evidence for suppression of smooth muscle contraction by a putative epithelium‐derived inhibitory factor (EpDIF) could be obtained: no inhibition of smooth muscle contractility was seen when the agents listed above were perfused intraluminally and their perfusion continued while they were applied to outside. 5 It was concluded that the epithelium plays a crucial role as an impermeant barrier in modulating the responsiveness of the airways smooth muscle.

Foetal airway motor tone in prenatal lung development of the pig

The terminal airways from embryonic lung in situ or as explants exhibit rhythmic spontaneous contractions. Our objective was to see whether narrowing responses of the airways occurred throughout the bronchial tree in the first trimester foetus and, if so, to characterize them. The bronchial tree was freed of vasculature and parenchyma from the lungs of 20-35 g pig foetuses (44-48 days gestation). The airway lumen was visualized directly with transmitted light, and narrowing was recorded in real time by video-imaging microscopy. From the main stem bronchi to the terminal regions of late generation branches (20-35 microns i.d.) strong bronchoconstrictor responses to micromolar concentrations of acetylcholine (ACh), histamine, substance P and K+ depolarizing solution were seen, whilst inhibition of narrowing with beta-adrenoceptor agonists was evidence of beta-receptors on the smooth muscle. Moreover, strong narrowing responses to electrical field stimulation, which were blocked by atropine, indicated that functional cholinergic nerves were present. A remarkable display of spontaneous narrowing in the airways of many of the bronchial tree preparations caused the movement of lung liquid to and fro. We speculate that the bronchomotor tone and associated spontaneous activity, which move the lung fluid along the airways, serve to maintain an even positive pressure in localized areas of the bronchial tree which is essential to provide the stimulus for continued growth of the lung.

Bronchial compliance and wall structure during development of the immature human and pig lung

Maturational changes in the specific compliance could potentially contribute to the development or clinical presentation of respiratory diseases in infants and children. Changes in the specific compliance during development and its structural basis have been well characterized, but changes in bronchial compliance and the mechanisms involved have received little attention. Semistatic pressure-volume curves were generated for isolated bronchial segments from late-term foetal, immature and adult pigs. A small number of bronchi from human infants were also studied. The amount of cartilage in the bronchial wall of pigs of different ages was measured histologically, and morphometric changes in the wall of inflated bronchi were investigated. The specific compliance of bronchi approximately halved from 1 to 4 weeks of age. No change in specific compliance was observed either between 4 week old and adult pigs, or between late-term foetal and 1 week old pigs. Changes in the total wall and cartilage areas did not correlate with changes in specific compliance. Inflation to 20 cmH2O transmural pressure reduced the total wall area of bronchi from 1 week old pigs. Significant changes in bronchial distensibility occur during the early postnatal period. These changes in specific compliance are not caused by an increase in the amount of cartilage. The increase in luminal volume during inflation of bronchial segments occurs, partially, by compression of the airway wall against the cartilage layer.

Airway narrowing and bronchodilation to deep inspiration in bronchial segments from subjects with and without reported asthma.

The present study presents preliminary findings on how structural/functional abnormalities of the airway wall relate to excessive airway narrowing and reduced bronchodilatory response to deep inspiration (DI) in subjects with a history of asthma. Bronchial segments were acquired from subjects undergoing surgery, mostly to remove pulmonary neoplasms. Subjects reported prior doctor-diagnosed asthma (n = 5) or had no history of asthma (n = 8). In vitro airway narrowing in response to acetylcholine was assessed to determine maximal bronchoconstriction and sensitivity, under static conditions and during simulated tidal and DI maneuvers. Fixed airway segments were sectioned for measurement of airway wall dimensions, particularly the airway smooth muscle (ASM) layer. Airways from subjects with a history of asthma had increased ASM (P = 0.014), greater maximal airway narrowing under static conditions (P = 0.003), but no change in sensitivity. Maximal airway narrowing was positively correlated with the area of the ASM layer (r = 0.58, P = 0.039). In tidally oscillating airways, DI produced bronchodilation in airways from the control group (P = 0.0001) and the group with a history of asthma (P = 0.001). While bronchodilation to DI was reduced with increased airway narrowing (P = 0.02; r = -0.64)), when the level of airway narrowing was matched, there was no difference in magnitude of bronchodilation to DI between groups. Results suggest that greater ASM mass in asthma contributes to exaggerated airway narrowing in vivo. In comparison, the airway wall in asthma may have a normal response to mechanical stretch during DI. We propose that increased maximal airway narrowing and the reduced bronchodilatory response to DI in asthma are independent.

Airway narrowing in porcine bronchi with and without lung parenchyma

During bronchoconstriction elastic after-loads arise due to distortion of lung parenchyma by the narrowing airway. In the present study, the functional effect of parenchymal elastic after-load on airway narrowing was determined. Airway narrowing was measured in vivo over a range of transpulmonary pressures and compared with in vitro narrowing measured at corresponding transmural pressures. Bronchi were generation 10 with internal diameters of ∼4 mm. In vivo luminal narrowing was measured by videobronchoscopy in anaesthetised and ventilated pigs. In vitro luminal narrowing was measured by videoendoscopy in isolated bronchial segments. Airways were activated by maximum vagal nerve stimulation and maximum electrical field stimulation in vivo and in vitro, respectively. At 5 cmH2O, stimulation produced a 35.9±3.2% (n = 6) and a 36.5±2.4% (n = 11) decrease in lumen diameter in vivo and in vitro, respectively. At 30 cmH2O, luminal narrowing fell to 23.7±2.0% in vivo and 23.4±2.5% in vitro. There was no difference between luminal narrowing in vivo and in vitro at any pressure. In conclusion, these findings suggest that in mid-sized, cartilaginous bronchi, parenchymal elastic after-loads do not restrict airway narrowing.

Increased responsiveness to cholinergic stimulation of small compared to large diameter cartilaginous bronchi

Model studies incorporating known morphological properties of the airways predict different narrowing characteristics between large and small bronchi. However, the flow-response characteristics of airways from different regions in the bronchial tree are still unknown. We compared the responsiveness of large and small bore bronchi of the pig to luminal or adventitial stimulation by acetylcholine (ACh) during perfusion of the airways at optimal driving pressure. Subsequently, each segment was measured morphometrically. Resting flow through the lumen was directly related to the fourth power of the segment internal diameter (ID). The sensitivity of small bronchi (2.00 mm ID) to ACh applied adventitially was about 870 times greater than large (5.85 mm ID) with a longer latency time of contraction in large bronchi. When perfused luminally the sensitivity difference was 42 fold, and after removing the epithelium the sensitivity difference was reduced to approximately five fold. Maximum concentrations of ACh produced airway closure in small segments (100% reduction in flow) but in large segments a plateau developed at 60-90% flow reduction. Similar findings were obtained with carbachol. Morphometry of the airway wall showed increasing cartilage, mucosal and smooth muscle thickness with increasing diameter, but a decreasing relative wall area. The marked differences in sensitivity to narrowing between large and small bronchi are, in part, related to differences in morphological properties of the mucosa (the epithelium) and adventitia, and to airway location. Airway size plays an important role in the maximum response of airway narrowing, with small airways showing closure and large airways a plateau.

Maintenance of airway caliber in isolated airways by deep inspiration and tidal strains

Deep inspirations (DIs) are large periodic breathing maneuvers that regulate airway caliber and prevent airway obstruction in vivo. This study characterized the intrinsic response of the intact airway to DI, isolated from parenchymal attachments and other in vivo interactions. Porcine isolated bronchial segments were constricted with carbachol and subjected to transmural pressures of 5-10 cmH2O at 0.25 Hz (tidal breathing) interspersed with single DIs of amplitude 5-20 cmH2O, 5-30 cmH2O, or 5-40 cmH2O (6-s duration) or DI of amplitude 5-30 cmH2O (30-s duration). Tidal breathing was ceased after DI in a subset of airways and in control airways in which no DI was performed. Luminal cross-sectional area was measured using a fiber-optic endoscope. Bronchodilation by DI was amplitude dependent; 5-20 cmH2O DIs produced less dilation than 5-30 cmH2O and 5-40 cmH2O DIs (P=0.003 and 0.012, respectively). Effects of DI duration were not significant (P=0.182). Renarrowing after DI followed a monoexponential decay function to pre-DI airway caliber with time constants between 27.4+/-4.3 and 36.3+/-6.9 s. However, when tidal breathing was ceased after DI, further bronchoconstriction occurred within 30s. This response was identical in both the presence and absence of DI (P=0.919). We conclude that the normal bronchodilatory response to DI occurs as a result of the direct mechanical effects of DI on activated ASM in the airway wall. Further bronchoconstriction occurs by altering the airway wall stress following DI, demonstrating the importance of continual transient strains in maintaining airway caliber.