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J Appl Physiol 87: 415-427, 1999;
8750-7587/99 $5.00
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Vol. 87, Issue 1, 415-427, July 1999

Surfactant effects in model airway closure experiments

K. J. Cassidy1, D. Halpern2, B. G. Ressler1, and J. B. Grotberg3

1 Department of Biomedical Engineering, Northwestern University, Evanston, Illinois 60208; 2 Department of Mathematics, University of Alabama, Tuscaloosa, Alabama 35487; and 3 Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109


    ABSTRACT
TOP
ABSTRACT
INTRODUCTION
PREVIOUS RESEARCH
EXPERIMENTS
EXPERIMENTAL RESULTS
LINEAR STABILITY ANALYSIS FOR...
THEORETICAL RESULTS
DISCUSSION
APPENDIX
REFERENCES

The capillary instability that occurs on an annular film lining a tube is studied as a model of airway closure. Small waves in the film can amplify and form a plug across the tube. This dynamical behavior is studied using theoretical models and bench-top experiments. Our model predicts the initial growth rate of the instability and its dependence on surfactant effects. In experiments, an annular film is formed by infusion of water into an initially oil-filled glass capillary tube. The thickness of the oil film varies with the infusion flow rate. The instability growth rate and closure time are measured for a range of film thicknesses. Our theory predicts that a thinner film and higher surfactant activity enhance stability; surfactant can decrease the growth rate to 25% of its surfactant-free value. In experiments, we find that surfactant can decrease the growth rate to 20% and increase the closure time by a factor of 3.8. Functional values of a critical film thickness for closure support the theory that it increases in the presence of surfactant.

pulmonary surfactants; surface tension; closing volume; airway liquid lining; microgravity


    INTRODUCTION
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ABSTRACT
INTRODUCTION
PREVIOUS RESEARCH
EXPERIMENTS
EXPERIMENTAL RESULTS
LINEAR STABILITY ANALYSIS FOR...
THEORETICAL RESULTS
DISCUSSION
APPENDIX
REFERENCES

THE LUNG'S AIRWAYS are liquid-lined flexible tubes. It is well recognized that the liquid lining of the lung can close off a small airway by forming a liquid plug and/or by provoking the collapse of the flexible airway wall (17, 32). The formation of a plug blocks airflow and can further induce the complete collapse of an airway, since liquid continuously drains into the plug. This is a postclosure filling flow, which extends the axial length of the plug. This happens most frequently near the end of expiration, when the airway diameters are small, i.e., at the closing volume. During inhalation the airways expand, which may result in the rupture of the plug and the reestablishment of airflow to the periphery (13, 23, 34). The closure and subsequent reopening of these airways contribute to the shape of the pressure-volume curve in cycled lungs, which characterizes the mechanical response of the lung. Understanding the phenomena that determine it is a fundamental inquiry in pulmonary physiology in normal and microgravity environments. Measuring the occurrence of airway closure is one component of a standard pulmonary function test: the single-breath nitrogen-washout test. When this test shows early airway closure, it is often interpreted that there is inhomogeneous ventilation distribution within the lung, which may be caused by a number of normal and pathological conditions (6). Surfactants tend to stabilize the lining from closure, as has been shown qualitatively (31).

There are likely several contributing forces that lead to airway closure. Studies by Halpern and Grotberg (20, 21) have modeled closure, including the effects of a flexible wall. We focus on modeling closure of smaller airways, typically beyond and including generation 16, in which gravitational effects are small. Our present study investigates one of the key mechanisms believed to cause airway closure: a surface tension-driven instability. By isolating the experimental system from gravitational and flexible-wall effects, we are able to focus on the effects of the surface tension instability on closure and the relative effects of surfactant.

A capillary instability occurs on the wavy interface between two fluids. A circular-cylindrical tube is lined with a uniform layer (the film) of viscous fluid surrounding a less viscous fluid (the core). It is well known that this configuration is unstable to finite-wavelength disturbances. We use a rigid-tube experimental model and compare the experimental results with rigid-tube theory. Depending on the initial film thickness, such disturbances may culminate in stable, steady, finite-amplitude waves or lead to a topological transition in which the film completely pinches off the core fluid, forming a liquid plug. We examine the effects of surfactant on the surface tension-induced instability that contributes to airway closure.

Glossary

We find it helpful to convert some dimensional variables into dimensionless variables. We use the following guide to define which are dimensional and dimensionless. Greek symbols representing the fixed fluid parameters (µ and rho ) are dimensional, and changing fluid parameters are dimensionless unless indicated with an asterisk. Roman variables representing the fixed system parameters (a, b, and g) are dimensional, and all other Roman variables are dimensionless unless indicated with an asterisk.

a Inner radius of the rigid tube
b Radius of the undisturbed cylinder of fluid (core radius)
C* Bulk concentration of surfactant
Ca µfilmU*/sigma *, capillary number for film formation
D*s Surface diffusivity of surfactant
D µfilmD*s/(h*0sigma *m<OVL>&dgr;</OVL>)
 &cjs1690;  epsilon -3D
 <OVL>&dgr;</OVL>  -(Gamma *m/sigma *m)dsigma */d&Ggr;*|&Ggr;*=&Ggr;*m, Marangoni number
 <A><AC>&dgr;</AC><AC>ˆ</AC></A>  epsilon -2<OVL>&dgr;</OVL>, surface activity parameter
 epsilon (a - b)/a, dimensionless thickness of the undisturbed film
 epsilon crit Critical dimensionless film thickness
 epsilon f Functional value of epsilon crit (taken at tc = 2 × 104)
g Gravitational constant
 Gamma * Surface concentration of surfactant
 Gamma *m Mean surface concentration of surfactant
 Gamma  Gamma */Gamma *m = 1 + epsilon 2<OVL>&Ggr;</OVL>
 <OVL>&Ggr;</OVL> Variation of surface concentration from the mean
 <A><AC>&Ggr;</AC><AC>ˆ</AC></A>  epsilon 2<OVL>&Ggr;</OVL>
h*0 Undisturbed film thickness
h* Amplitude of the film disturbance
h h*/epsilon a
h1 Initial amplitude (perturbation parameter)
k* 2pi /lambda *, wave number of the disturbance
k k*a
K k*b
k*1 Surfactant adsorption constant
k*2 Surfactant desorption constant
 lambda * Wavelength of the disturbance
 lambda  lambda */a
µcore Viscosity of the core fluid
µfilm Viscosity of the film fluid
q* Initial growth rate of the disturbance
q q*aµfilm/sigma *
&qcirc; q*epsilon -3aµfilm/sigma *
r r*/a, radial distance from the tube axis
 rho core Density of the core fluid
 rho film Density of the film fluid
 sigma * Surface tension at the interface between the fluids
 sigma *m Mean value of surface tension
t*f Final time of the disturbance (at closure)
t*0 Start time of the disturbance
t*c t*f - t*0, total time for closure to occur
t t*/(aµfilm/sigma *)
t t*/(epsilon -3aµfilm/sigma *)
U* Mean velocity of the infused core fluid (during film formation)
z z*/a, distance along the tube axis


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APPENDIX
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Mechanical phenomena related to airway closure can be drawn from several sources. From the physiological literature, the stabilizing effects of surfactants on models of airway closure have been demonstrated. Liu et al. (31) conducted experiments where air is forced through a narrow glass or esophageal tube segment initially filled with saline or a surfactant-saline solution. They used a constant tube size and driving pressure, with a constant liquid volume localized to the narrow, constricted segment of the tube. The liquid in the tube gives way to airflow but reforms a plug in systems with little surfactant or surfactant weakened by albumin and fibrinogen. However, the plug does not reform if adequate surfactant is present, for the conditions explored. This demonstrates the important clinical effects of exogenous surfactant replacement therapy (11, 30, 44), which can help stabilize airways in patients who have a surfactant deficiency. Liu et al. attributed this additional stability to the reduction of surface tension caused by the surfactant polar molecules. Saline lung lavage in isolated rat lungs reduces the endogenous surfactant concentration, causing an increase in airway resistance due to rapid plug formation (7, 8). However, instillation of exogenous calf lung surfactant extract after the lavage can increase the duration of unobstructed airway time by a factor of 3 (8) to 7 (7), determined by measuring resistance via airway pressure. It would be interesting to examine the airway closure and film amplification for a range of initial film thicknesses and compare behavior before and after addition of exogenous surfactant.

From the engineering literature, the study of capillary instabilities, such as the breakup of a cylindrical column of fluid or a thin film coating a rigid tube, continues to be an area of ongoing investigation. Plateau (37) considered the static stability of a cylindrical column of fluid with undisturbed radius b. He proved, using energy arguments, that a perturbation to the interface is unstable if its wavelength (lambda *) exceeds the circumference of the column (2pi b). The perturbation causes axial waves in the interface, similar to waves in the interface of our system, which we show and discuss in Experimental methods. Rayleigh (40) studied the dynamic stability of an initially cylindrical liquid jet. He found that waves at the surface amplify until the jet breaks up into a stream of droplets, provided Plateau's criterion holds, i.e., that K < 1, where K = k*b = 2pi /lambda * is the disturbance wave number. Rayleigh derived a relationship between the disturbance growth rate (q*) and K. The q* attains a maximum at some critical wave number (Kcrit). He found the least-stable mode at Kcrit = pi /4.5 approx  0.698 for a liquid jet in an inviscid or a viscous fluid. Rayleigh (41) modified this analysis for a jet of air immersed in a liquid and found the same criterion for the maximum growth rate (qmax): Kcrit = pi /4.5.

Goren (16) studied the stability of an annular liquid film lining a tube otherwise filled with air. A rigid tube of radius a, lined with a uniform film at radial distance b, has a dimensionless film thickness epsilon  = (a - b)/a. A disturbance of lambda * causes axial waves in the interface, which can amplify and eventually cause film closure. Goren performed experiments for 0.06 < epsilon  < 0.33 and allowed the systems to reach closure. Plug formation occurred at multiple, spatially periodic locations defined by Kcrit, which was shown to depend weakly on epsilon . In his accompanying theoretical analysis for the initial growth rate, q = q*aµfilm/sigma *, (where µfilm is viscosity of the fluid film), he predicts how qmax depends on Kcrit and epsilon . Goldsmith and Mason (15) performed closure experiments using immiscible liquids of similar density for the film and core fluids to minimize buoyancy. They measured disturbance wave numbers that match Goren's theory well and initial growth rates that were slightly greater than predicted by theory (16). Sample results are shown tracing the logarithm of the amplitude of the growth vs. time for two experiments, at epsilon  = 0.19 and epsilon  = 0.25, on a surfactant-free system. The disturbance amplitude initially grows exponentially with time, at a constant q, but as the wave amplifies, q accelerates. Also, the closure times (tc) of the two sample experiments can be depicted from this graph.

The epsilon  must be larger than a critical value (epsilon crit) for closure to occur. Disturbance waves in a film that is thinner than epsilon crit may amplify to form undulate collars spaced throughout the tube. However, not enough liquid volume is present for the collars to bridge the tube. Everett and Haynes (9) found that a collar of surfactant-free fluid wetting a rigid tube would undergo a topological transformation to a liquid plug, provided the collar volume exceeds 5.47a3 enclosed within one axial wavelength. Similar results were also obtained by Kamm and Schroter (28), who conducted experiments where oil is dripped into small vertical rigid tubes. By assuming an initial film thickness of 10 µm at total lung capacity (TLC), they estimated that airway closure first occurs in the terminal bronchioles (about generations 16-17) at a lung volume of 23% TLC. For an initial film thickness of 5 µm, airway closure first occurs at 12% TLC (28). These closing volumes are commonly seen in pulmonary function tests.

Gauglitz and Radke (12), extending theoretical work by Hammond (22), computed an epsilon crit = 0.12 necessary for closure, whereas their experiments indicate an average epsilon crit = 0.09. Johnson et al. (27) analyzed the stability of a viscous annular film, including the effects of inertia. They found their approximation of the initial growth rate to match Goren's exact solution well. Closure of liquid-lined, linearly elastic tubes has been investigated by Halpern and Grotberg (20). They found that flexibility enhances the growth of the unstable film. Theoretical models developed to examine the effects of surfactant on film stability (21, 35) show that plug formation in a system containing surfactant is slowed by a factor of ~4 over the surfactant-free model. Halpern and Grotberg (21) found that epsilon crit is increased in a system containing surfactant, and epsilon crit decreases with increasing ratio of surface forces to elastic forces.

In EXPERIMENTS, we describe the materials and methods for performing experiments on annular film closure in a capillary tube. We consider a range of initial film thicknesses and examine systems without and with surfactant. The experimental system uses liquids for the core and film fluids that approximately match the viscosity ratio and Reynolds numbers (Re) in the lung system. Gravitational effects are negligible, inasmuch as the density difference between the core and film liquids is small. This allows us to investigate the closure phenomena induced by nongravitational forces. Also, film closure occurs much more slowly in a liquid-liquid system than in an air-liquid system, allowing the film dynamics to be viewed and recorded in real time. Then we report the experimental results of tc, q, and epsilon crit; we also report inflow results of the annular film formation. Next we outline our theoretical model for the initial growth rate in a system containing surfactant and show theoretical results. Then we discuss some observed behavior, compare the experimental data with theory, and make conclusions about the effects of surfactant on film formation and stability.


    EXPERIMENTS
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Materials. Four fluid systems are used in these sets of experiments (Table 1). We use high-viscosity oils as the film fluid and water as the core fluid. In systems 1 and 2, the oil used for the annular film is LB-1715 (Union Carbide Chemical, South Charleston, WV), which is a propylene glycol monobutyl ether. System 1 is surfactant free, and system 2 contains a small amount of the surfactant 1-palmitoyl-2-lauroyl-sn-glycero-3-phosphocholine (Avanti Polar Lipids, Alabaster, AL; PLPC). It is an asymmetric fatty acid that is insoluble in water. System 2 contains 10-3 mM bulk concentration of PLPC dissolved in the oil phase before contact with the water phase. In systems 3 and 4, the oil used for the annular film is LB-400-X (Union Carbide Chemical). System 3 is surfactant free, and system 4 contains a small amount of the surfactant Tween 20 (Sigma Chemical, St. Louis, MO). Tween 20 is polyoxyethylenesorbitan monolaurate and is made up of various fatty acids; it is soluble in water and insoluble in oil. It is initially dissolved in the water phase at a bulk concentration of 0.01% by weight. In Table 1 the film viscosities were measured at room temperature (24-25°C) with an Oswald bulb viscometer, and the film densities are from manufacturer specifications. The surface tension of the LB-1715 and LB-400-X in air is 33-38 dyn/cm. Mean values for the surface tension of an oil-water interface were measured using a ring tensiometer and are displayed in Table 1. In calculations that require a value for the surface tension (sigma *) for these experiments, we use the mean values for each of the four systems.

                              
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Table 1.   Properties of the experimental systems

Similitude: matching parameters. The system parameters are organized into dimensionless groups (Table 2). We attempt to model the ranges found in the pulmonary airways, with some important differences. We assume that at generation 16 in normal lungs the film is the aqueous phase with a mean airway diameter of 0.049 cm (46). In calculating lung properties in Table 2, we use the density and viscosity values of water and air for the film and core fluids, respectively. The surface tension varies by region of the lung because of the local quantity of surfactant and by lung volume because of dynamic surface concentration of surfactant. The tracheal surface tension was directly measured to be 31 dyn/cm in normal horse lungs (24) and 33 and 32 dyn/cm in normal rat and guinea pig lungs, respectively (29). The equilibrium surface tension in the alveolar region of rabbit lungs was measured to be 7 and 30 dyn/cm at 50 and 100% TLC, respectively (43). We expect the surface tension in normal pulmonary airways to be ~20-25 dyn/cm at functional residual capacity, which agrees with previously published values (14, 18). We use sigma * = 20 dyn/cm in Table 2 for calculations of normal lung parameters at generation 16. 

                              
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Table 2.   Dimensionless parameters for the pulmonary system and the experimental models

In normal lungs the film lining is very thin, but many diseases as well as the state of lung inflation can affect epsilon , the ratio of film lining thickness to airway radius. Yager et al. (48) measured the airway surface liquid thickness in the small airways of the guinea pig. With use of data from their Fig. 4 on the airway perimeter and film thickness, and the assumption that the airways are approximately circular in cross section, the average ratio of film thickness to airway radius is 0.02. In a study by Codd et al. (4), in a sheep trachea, the mean epsilon  = 0.04. Excessive bronchial secretions in individuals with unhealthy lungs can commonly produce airway surface liquid films that are 10% of the airway diameter (42); this corresponds to epsilon  = 0.20. In our experiments we model airways in which the film is thick enough to close: epsilon  > 0.12.

The Bond number (Bo) = Delta rho g a2/sigma * (where rho  is fluid density and g is gravitational constant) indicates where gravitational effects are significant. Here, Delta rho  = |rho film - rho core|, the difference in densities between the two fluids. Gravity can be neglected, provided epsilon -1 Bo << 1 (22), which generally holds in smaller airways. Under normal-gravity conditions, for epsilon  = 0.15, we calculate that epsilon -1 Bo < 0.3, starting at generation 16 in adults and starting at generation 9 in infants. In the case of lessened gravity (such as the microgravity environment) or higher surface tension (which can occur in premature infants born with insufficient pulmonary surfactant) the Bo values would be smaller. In our experiments we wish to examine the dynamics of closure as contributed by forces other than gravitational. By using a liquid-liquid system, where the fluid densities are similar, we can create systems with low Bo values by reducing Delta rho . The time scale of these experimental systems is larger than the air-liquid system in the airways by a factor of 104. By slowing the closure process, data capture is simplified, with improved accuracy in the results. These fluids are chosen, oil for the film and water for the core fluid, such that the viscosity ratio gamma  = µcorefilm << 1, which matches the lungs. Fluid inertia is negligible, provided epsilon 2 Refilm = (epsilon 5rho filmsigma *a)/µ2film << 1 (22), which is true in the lungs (Table 2). Similarly, fluid inertia in the core is neglected, since epsilon 2 Recore = [(epsilon 5sigma *)/µfilm][(rho corea)/µcore] << 1, as in the lungs.

To create a surfactant monolayer between the two fluid layers, we first dissolve the surfactant into one of the bulk fluids. When the interface between the two fluids is formed, we assume that the surfactant rapidly adsorbs to the surface and does not rapidly desorb back into the bulk of the fluid. By calculating the solubility parameter beta  = (k*1C*)/(k*2h*0), we can estimate the ratio of time scales for desorption to adsorption (26), where k*1 is the adsorption coefficient, k*2 is the desorption coefficient, C* is the bulk concentration of surfactant, and h*0 is the film thickness. Jensen and Grotberg (26) estimate that the pulmonary surfactant component dipalmitoylphosphatidylcholine (DPPC) dissolved in an aqueous lung surface layer 1 µm thick will have beta  = 5 × 107. At generation 16 the airway surface liquid is ~6-12 µm thick (using 0.02 <=  epsilon  <=  0.03), yielding 4.2 × 106 <=  beta  <=  8.5 × 106. The surfactant PLPC used in our experiments has adsorptive/desorptive properties similar to DPPC. Using the range of film thicknesses found in our experiments (3.5 µm <=  h*0 <=  10.4 µm), we estimate 1.4 × 106 <=  beta  <=  4.8 × 106 for the system containing PLPC. Because beta  >> 1 in the lung and in our experiments, we assume that surfactant concentration at the interface is initially uniform. Surface diffusivity of surfactant is measured by the diffusivity parameter &cjs1690; = epsilon -3D, where D = -[(µfilmD*s)/(h*0sigma *m<OVL>&dgr;</OVL>)] and D*s is the dimensional surface diffusivity. The Marangoni number, defined as <OVL>&dgr;</OVL> = -(Gamma *m/sigma *m)dsigma */d&Ggr;*|&Ggr;*=&Ggr;*m, is a measure of the ability of the surfactant to lower the surface tension of a monolayer. Here, Gamma *m is the mean value of Gamma *, the surface concentration of surfactant, and sigma *m is the mean value of sigma *. Agrawal and Neuman (1) directly measured the surface diffusivity of DPPC in myristic acid and found 10-7 < D*s < 7 × 10-5 cm2/s. Jensen and Grotberg (25) approximate that D is on the order of 10-6 for experiments and 10-4 in an airway. For epsilon  = 0.1, this still makes &cjs1690; quite small: 10-3 <=  &cjs1690; <=  10-1. Therefore, the current theory neglects surface diffusivity.

Experimental methods. A thin cylindrical borosilicate glass capillary tube with an inner diameter of 0.058 cm and length of 20 mm is viewed under a microscope-camera configuration (Fig. 1). The glass capillary is cleaned with chromic acid, rinsed with acetone and deionized water, and dried before the experiment. Fluids are delivered to the capillary tube through flexible Teflon tubing having the same inner and outer diameters as the rigid capillary tube and held together by a sleeve of silicone tubing and glue. A three-way stopcock is used to direct the fluids from respective syringes through the Teflon tubing. The film fluid is injected manually, and a positive-displacement syringe pump (model 341-A, Sage Instruments) drives the core fluid through the tube. The capillary tube, immersed in glycerol and held between two glass slides perpendicular to the microscope to correct for visual distorsion, is then placed under the microscope and into a pipette holder, which is attached to a Leitz micromanipulator. A camera is attached to the top of a Leitz Laborlux D microscope; the capillary tube is viewed at ×200 or ×500 magnification. All experiments are recorded from a Javelin JE3362 camera connected to a JVC SR-S360 video recorder. A Sony PVM-2030 20-in. color monitor is also connected, which provides for viewing capabilities during experiments and later for data acquisition.


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Fig. 1.   Schematic of experimental apparatus.

To measure the value of the mean interfacial surface tension between the film fluid and the core fluid, we use a standard 6-cm platinum-iridium ring tensiometer. Measuring accurately the surface tension between liquids of similar densities can be difficult. In the tensiometer test, a thin ring is pulled up through the horizontal liquid-liquid interface, creating a vertical force on the ring, which is then measured by the tensiometer scale. However, a ring pulled through liquids of similar density may not produce a vertical interface, particularly on the inner edge of the ring. The inaccuracy may be quite large, since the ring tensiometer measures only the vertical component of the force. For contact angles deviating from vertical by up to 30°, the true surface tension may be underestimated by as much as 15%. Assuming a vertical interface, we estimate the mean surface tension of LB-1715 with deionized distilled water to be 4.59 dyn/cm. The addition of the surfactant PLPC added to the oil phase in molar bulk concentrations of 10-4-10-1 mM causes a slight decrease in the mean surface tension to 4.23 dyn/cm (<8% change). The mean surface tension of LB-400-X and water, estimated at 4.77 dyn/cm, exhibits a dramatic decrease with the addition of 0.01% (by weight) Tween 20, to 2.88 dyn/cm (~40%).

As part of the preliminary steps in performing stability experiments, it is necessary to create the two-phase system of an annular film lining a tube with a fluid core. This system is established by first filling the tube with the film fluid from a 10-ml syringe. The core fluid is then pumped into the center of the tube, driven by a positive-displacement syringe pump. This core fluid flows through the center of the tube like a long finger (Fig. 2A) and leaves an axisymmetric annular film on the tube walls (Fig. 2B). The thickness of the film depends on the capillary number (Ca = µfilmU*/sigma *, where µfilm is the film viscosity, U* is the average core fluid velocity, and sigma * is the interfacial surface tension). Previous studies (10, 15, 45) support an empirical equation for epsilon  
&egr; = 0.5Ca<SUP>½</SUP> (1)
for 10-4 < Ca < 10-1. Results from several experimental studies support Bretherton's theoretical analysis (2), indicating
&egr; = 0.643(3Ca)<SUP>2/3</SUP> (2)
for 10-4 < Ca < 10-2. Martinez and Udell (33) conducted a numerical analysis on a finite-length bubble to describe the shape at the leading and trailing menisci. Their predictions of epsilon (Ca) are in excellent agreement with experimental data by Taylor (45) for 10-2 <=  Ca <=  2 and support Eq. 1 for 10-2 <=  Ca <=  10-1. Goldsmith and Mason (15) also performed experiments on two sets of fluid systems with and without the surfactant Tween 20. Without surfactant, they concluded that epsilon  varies like Ca0.5 for small Ca. However, when 0.5% Tween 20 is added to the water phase of the two systems, they find that epsilon  varies like Ca0.28 and Ca0.44, respectively.


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Fig. 2.   Schematic of inflow (A), uniform annular film (B), and disturbed film (C). See Glossary for definition of abbreviations.

The average core fluid velocity is defined as the infusion flow rate divided by the measured cross-sectional area of the core. Using this relationship, we can directly control the formed film thickness by varying the flow rate settings on the syringe pump. In creating the annular film, syringe pump settings range from 0.35 ml/h to 0.49 ml/min. Here, films are formed with epsilon  ~0.12-0.36. It is necessary to record epsilon  before waves develop. Measurements of epsilon  are taken at several locations between about two and five tube diameters from the leading meniscus of the infused core fluid, then averaged. Once the uniform film is established throughout the tube, the inflow is stopped and the downstream end of the tube is plugged. The tube is then scanned to search for small waves at the interface with use of a finely controlled micromanipulator, and wave amplification (Fig. 2C) is viewed and recorded.

Figure 3 shows six photographs taken from a sample experiment in its entirety; Fig. 3, A and B, are from the inflow part; Fig. 3, C-F, correspond to the film instability. We use Targa-Plus software and Frame-Grabber PC board. In Fig. 3A the film fluid is displaced by a finger of the core fluid, which leaves a uniform annular film, as shown in Fig. 3B. The infusion flow is shut off, then a small disturbance is located (Fig. 3C), which then amplifies (Fig. 3D). Measurements of the film interface evolution are taken at the longitudinal position where the radius of the interface is the smallest. Interface positions are recorded periodically, and the time at closure is noted. We see the film as it amplifies just before closure in Fig. 3E, then just after closure in Fig. 3F. The recorded data are analyzed to determine the initial, constant growth rates and the closure times as a function of epsilon  and system parameters. For this sample experiment shown on system 2, recorded values are as follows: inflow rate = 2.7 ml/h, epsilon  = 0.29, q* = 0.155 s-1, and closure time (t*c) = 24 s.



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Fig. 3.   Photographs of experimental procedure. Inflow begins (A) and a uniform film is established (B); then inflow is stopped, a small disturbance is located (C), and disturbance amplifies (D). E: just before closure. F: just after closure.

Determining t*c and initial growth rate. To extract t*c from an experimental trial, it is necessary to determine the start time (t*0) of the disturbance. However, a wave has already begun to amplify (t* > t*0) before it is large enough to be visually detected. Below, we describe a method that allows the dimensional values of t*0 and the initial growth rate q* to be estimated. A small wave disturbance initially grows exponentially with time. A normal mode expression for h = h1eq*t*+ik*z* is used where h = h*/epsilon a is a sinusoidal film amplitude. By plotting ln(h) vs. t* from our data, the abscissa represents an arbitrary time corresponding to the time of the videotape recording. A straight-line curve fit to the early data results in the t* = 0 intercept at h = h1. The t*0 of the perturbation is taken to be the time at which the disturbance amplitude is 5% of the undisturbed film thickness. The slope of the initial portion of this graph also gives q* (in s-1). Then t*c = t*f - t*0, where t*f is the final time documented at closure (in s). Values for the dimensionless growth rate and closure time are then computed.


    EXPERIMENTAL RESULTS
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ABSTRACT
INTRODUCTION
PREVIOUS RESEARCH
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THEORETICAL RESULTS
DISCUSSION
APPENDIX
REFERENCES

In Figs. 4-6, involving our experimental data, symbols represent average values for a set of experiments. A "set" constitutes a group of film systems that were created by using a specific inflow rate to provide consistent values of epsilon  for the set. There is some deviation within a set for epsilon , as well as the values of tc [t*c/(aµfilm/sigma *)], q [q*aµfilm/sigma *], and Ca. For this reason, bidirectional scatter bars, which are based on the standard deviation of the set, are shown. We used 13 flow settings in the range 0.35 ml/h to 0.49 ml/min. For each set, up to 20 experimental trials were performed and averaged. For the experiments on stability, useful data were extracted from two to eight trials per set. For inflow experiments establishing the uniform film thickness, the number of trials providing useful data varied from 2 to 20 per set.

Closure time and minimum film thickness. Figure 4 indicates how tc decreases as epsilon  increases and how the presence of surfactant delays closure. In Fig. 4A, the addition of PLPC to this system increases tc by up to a factor of 3.8 (the value at epsilon  = 0.19). Note that for increasingly thick films, the difference between the surfactant-free and the surfactant data becomes less significant. In Fig. 4B, the addition of Tween 20 causes an increase in tc by up to a factor of 3.0 (at epsilon  = 0.24). The thinnest films we found to close were at epsilon  = 0.123 for system 1, at epsilon  = 0.154 for system 2, at epsilon  = 0.143 for system 3, and at epsilon  = 0.156 for system 4.



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Fig. 4.   Closure time [tc = t*c/(aµfilm/sigma *)] vs. undisturbed film thickness (epsilon ) for systems 1 (bullet ) and 2 (open circle ; A) and systems 3 (bullet ) and 4 (open circle ; B). Surfactant delays closure, and thicker films take longer to close than thinner films. Note how closure time tends to infinity as epsilon  approaches epsilon crit. See text for theoretical curves.

The solid lines in Fig. 4 are computed from the surfactant-free theory of Halpern and Grotberg (20) in the rigid-tube limit. The dashed lines are computed from the theory of Halpern and Grotberg (21) that predicts closure in a system containing surfactant. We simultaneously solve their Eq. A1 numerically. Values are chosen for the surface activity (<OVL>&dgr;</OVL> = 8) and for the disturbance wavelength (lambda  = 2<SUP><FR><NU>3</NU><DE>2</DE></FR></SUP>pi ) corresponding to a system with normal surface activity and a disturbance with the most dangerous wavelength. We calculated epsilon crit = 0.13 for a surfactant system using these values (20).

Initial growth rate of the disturbance. The q of the capillary instability is found to increase as epsilon  increases and to decrease in the presence of surfactant. In Fig. 5A, the presence of PLPC in system 2 decreases the growth rate to as little as 21% (at epsilon  = 0.19) of surfactant-free system 1 values. In Fig. 5B, Tween 20 causes q to decrease to as little as 24% (at epsilon  = 0.20) of the corresponding surfactant-free values for these experiments.



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Fig. 5.   Initial growth rate (q = q*aµfilm/sigma *) vs. epsilon  for systems 1 (bullet ) and 2 (open circle ; A) and systems 3 (bullet ) and 4 (open circle ; B). Surfactant decreases growth rate, and thicker films grow faster than thinner films. See text for theoretical curves.

In Fig. 5, the solid lines are a reproduction of Goren's theory (16) for the maximum growth rate (qmax) of a surfactant-free system of arbitrary thickness epsilon . We have calculated the theory line by rescaling the wave number on the tube radius, or k = k*a. The three broken lines are from our linear theory of qmax, which is valid for thin films; we have shown lines representing the theory up to epsilon  = 0.20. The short-dashed line is our current theory in the limit of no surfactant, or <OVL>&dgr;</OVL> = 0. Derivation of our present theory is detailed in LINEAR STABILITY ANALYSIS FOR THE AIRWAY CLOSURE MODEL WITH SURFACTANT. Details on the theoretical results follow the derivation in THEORETICAL RESULTS. The long-dashed and long-and-short-dashed lines show our theory for an interface with a small amount of surfactant (<OVL>&dgr;</OVL> = 0.01) and saturated with surfactant (<OVL>&dgr;</OVL> = infinity ), respectively. Normal surface activity <OVL>&dgr;</OVL> = 8 lies in the narrow margin between the very small and very large surfactant activity values. As the surface activity increases, the growth rate decreases. Note how even a small amount of surfactant can cause a significant decrease in the growth rate compared with the same system without surfactant.

Thickness of the annular film. The epsilon  of the formed annular film is shown to increase with Ca for the inflow procedure in Fig. 6. In Fig. 6A, some experimental results of Goldsmith and Mason's surfactant-free system 6a (15) are also shown for comparison. There is a smooth transition between their data and ours. We find the asymptotic behavior as predicted by others (5, 33), with our experimental value near epsilon  = 0.34 at Ca = 10. In Fig. 6B, at a given Ca, epsilon  decreases slightly in the presence of Tween 20, from system 3 to system 4. However, the change in epsilon  with the addition of surfactant is comparable to the experimental scatter, which is <10%. The long-dashed line is Eq. 2, Bretherton's theory (2), and the short-dashed line is Eq. 1, from Fairbrother and Stubbs (10). Equation 2 is inaccurate for Ca > 10-2 and overpredicts epsilon  for all our experiments in this study. Equation 1 matches epsilon  well for Ca < 0.2 but is invalid for large Ca and overpredicts epsilon  for Ca > 0.2. We reproduce the numerical results of Martinez and Udell (33) (solid line) for a surfactant-free system, in the range 10-2 < Ca < 10. There is little variation in epsilon  between systems 1 and 2, the deviation being comparable in size to the experimental scatter, which ranges from 2 to 13%.



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Fig. 6.   epsilon  = (a - b)/a vs. capillary number (Ca = µfilmU*/sigma *) of infused fluid for systems 1 (bullet ) and 2 (open circle ) and Goldsmith and Mason's system 6a (black-down-triangle ; A) and systems 3 (bullet ) and 4 (open circle ; B). epsilon  increases with Ca, with little variation between surfactant-free and surfactant systems. See text for theoretical curves.


    LINEAR STABILITY ANALYSIS FOR THE AIRWAY CLOSURE MODEL WITH SURFACTANT
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A linear stability analysis is performed on the airway closure model with surfactant. The model is based on the inertia-free analysis developed by Halpern and Grotberg (20), who derived a system of nonlinear evolution equations (Eq. A1) for the film thickness and surface concentration of surfactant (see APPENDIX). To investigate linear stability of the system, we use normal-mode sinusoidal disturbances
<IT>h</IT> = <IT>h</IT><SUB>1</SUB><IT>e</IT><SUP><IT><A><AC>q</AC><AC>ˆ</AC></A><A><AC>t</AC><AC>ˆ</AC></A></IT> + <IT>ikz</IT></SUP> and <A><AC>&Ggr;</AC><AC>ˆ</AC></A> = <A><AC>&Ggr;</AC><AC>ˆ</AC></A><SUB>1</SUB><IT>e</IT><SUP><IT><A><AC>q</AC><AC>ˆ</AC></A><A><AC>t</AC><AC>ˆ</AC></A></IT> + <IT>ikz</IT></SUP> (3)
where &qcirc; = -q*epsilon -3aµfilm/sigma * and t = t*/(epsilon -3aµfilm/sigma *). The dimensionless surface concentration Gamma  = Gamma */Gamma *m = 1 + epsilon 2<OVL>&Ggr;</OVL>, where <A><AC>&Ggr;</AC><AC>ˆ</AC></A> = epsilon 2<OVL>&Ggr;</OVL> and <OVL>&Ggr;</OVL> is the variation of surface concentration from the mean. For Eq. 3, |h1| << 1 and ‖<A><AC>&Ggr;</AC><AC>ˆ</AC></A><SUB>1</SUB>‖ &z.Lt; 1, and substitute Eq. 3 into Eq. A1. We define the dimensionless wave number k = k*a and the surface activity parameter <A><AC>&dgr;</AC><AC>ˆ</AC></A> = epsilon -2<OVL>&dgr;</OVL>. The dispersion relation for &qcirc; is
<IT><A><AC>q</AC><AC>ˆ</AC></A></IT> = <FR><NU><IT>k</IT><SUP>2</SUP></NU><DE>6</DE></FR> <FENCE>1 − 3<A><AC>&dgr;</AC><AC>ˆ</AC></A> − <IT>k</IT><SUP>2</SUP> + <RAD><RCD><FENCE><FENCE><IT>k</IT><SUP>2</SUP> + <FR><NU>3</NU><DE>2</DE></FR> <A><AC>&dgr;</AC><AC>ˆ</AC></A> − 1</FENCE><SUP>2</SUP> + <FR><NU>27</NU><DE>4</DE></FR> <A><AC>&dgr;</AC><AC>ˆ</AC></A><SUP>2</SUP></FENCE></RCD></RAD></FENCE> (4)
If <A><AC>&dgr;</AC><AC>ˆ</AC></A> = 0, the constant surface tension result is recovered
<IT><A><AC>q</AC><AC>ˆ</AC></A></IT> = <FR><NU><IT>k</IT><SUP>2</SUP></NU><DE>3</DE></FR> (1 − <IT>k</IT><SUP>2</SUP>) (5)
Equation 5 can also be obtained by using normal modes with Hammond's analysis (22) or in the rigid-tube limit of Halpern and Grotberg (20). In the limit <A><AC>&dgr;</AC><AC>ˆ</AC></A> right-arrow infinity  
<IT><A><AC>q</AC><AC>ˆ</AC></A></IT> = <FR><NU><IT>k</IT><SUP>2</SUP></NU><DE>12</DE></FR> (1 − <IT>k</IT><SUP>2</SUP>) (6)
This limit can also be obtained by applying a no-slip condition at the fluid interface. Hence, in relatively large concentrations, surfactant slows closure by up to a factor of 4. This is consistent with results of Otis et al. (35), who discussed these two limits.


    THEORETICAL RESULTS
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Figure 7A shows &qcirc;max vs. <A><AC>&dgr;</AC><AC>ˆ</AC></A>. This comes from Eq. 4 by use of the corresponding most dangerous wave number kcrit at each value of <A><AC>&dgr;</AC><AC>ˆ</AC></A>. The &qcirc;max decreases with increasing surface activity. In the limit of vanishing surface activity, the growth rate approaches the constant surface tension value, &qcirc;max = <FR><NU>1</NU><DE>12</DE></FR>. As the interface becomes saturated with surfactant, the growth rate asymptotes to one-fourth of the surfactant-free value. This corresponds to the rigid-surface limit. Figure 7B shows kcrit vs. <A><AC>&dgr;</AC><AC>ˆ</AC></A>; kcrit varies weakly with <A><AC>&dgr;</AC><AC>ˆ</AC></A>. In the limit of zero surface activity, kcrit = 2-1/2 is recovered. There is a minimum value of kcrit at <A><AC>&dgr;</AC><AC>ˆ</AC></A> = 0.2, and the critical wave number then asymptotes to kcrit = 2-1/2 for an interface saturated with surfactant. Theoretically, typical <OVL>&dgr;</OVL> values range from 0 (no surfactant) to 8 (normal surfactant activity); <OVL>&dgr;</OVL> right-arrow infinity  represents an interface saturated with surfactant. With the assumption that epsilon  = 0.1-0.4, the corresponding values of <A><AC>&dgr;</AC><AC>ˆ</AC></A> range up to 800 (for 0 <=  <OVL>&dgr;</OVL> <=  8). In Fig. 7, for <A><AC>&dgr;</AC><AC>ˆ</AC></A> > 1, kcrit and &qcirc;max rapidly approach their respective asymptotic limits for an interface saturated with surfactant.




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Fig. 7.   Linear film stability, including surfactant effects. Maximum growth rate (&qcirc;max) vs. surfactant activity (<A><AC>&dgr;</AC><AC>ˆ</AC></A>) (A), critical wave number (kc) vs. <A><AC>&dgr;</AC><AC>ˆ</AC></A> (B), and &qcirc; vs. k (C) for (top to bottom) <A><AC>&dgr;</AC><AC>ˆ</AC></A> = 0, 0.1, 1, and infinity , respectively.

Figure 7C shows &qcirc; vs. k for <A><AC>&dgr;</AC><AC>ˆ</AC></A> = 0, 0.1, 1, and infinity . The &qcirc;max decreases with increasing surface activity. Because the growth rate appears to be more sensitive to changes in the <A><AC>&dgr;</AC><AC>ˆ</AC></A> at small values of <A><AC>&dgr;</AC><AC>ˆ</AC></A>, we focus on smaller values of <A><AC>&dgr;</AC><AC>ˆ</AC></A> for demonstration. The kcrit at which the growth rate is maximum shifts slightly as <A><AC>&dgr;</AC><AC>ˆ</AC></A> changes, according to Fig. 7B. At any given wave number, the growth rate of a disturbance is larger in a surfactant-free system than in a system with nonzero surface activity.


    DISCUSSION
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General observations. The growth of the disturbance is related to the epsilon  in general and to the relation of epsilon  to epsilon crit in particular. Figure 8 shows sample data from three different experiments. We observe that for epsilon  > epsilon crit (Fig. 8A; epsilon  = 0.152 compared with predicted epsilon crit = 0.12), the disturbance initially grows at a constant rate, which then accelerates until closure. When the disturbance amplifies, it draws fluid from the thinning region near the wall into the bulge of the film. For thin films where epsilon  is just slightly larger than epsilon crit (Fig. 8B; epsilon  = 0.131, predicted