Mucociliary interaction in vitro: effects of physiological and inflammatory stimuli

1990 ◽  
Vol 68 (4) ◽  
pp. 1421-1426 ◽  
Author(s):  
Z. V. Seybold ◽  
A. T. Mariassy ◽  
D. Stroh ◽  
C. S. Kim ◽  
H. Gazeroglu ◽  
...  
Keyword(s):  
Liquid Velocity ◽  
Beat Frequency ◽  
Platelet Activating Factor ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Base Line ◽  
Mucociliary Transport ◽  
Epithelial Surface ◽  
Surface Liquid

Mucociliary transport in the airways is governed by the interaction between ciliary activity and the depth and rheological properties of the liquids (mucus) covering the epithelial surface. A change in one of these parameters may not predict the direction and magnitude of a concomitant change in mucociliary transport. We therefore determined the effects of physiological (neurotransmitters) and pathological (inflammatory mediators) stimuli on ciliary beat frequency (CBF), surface liquid velocity (SLV), surface liquid depth (SLD), and viscoelasticity of mucus in pieces of sheep trachea (n = 5 for each treatment) mounted in a chamber such that the submucosal side was bathed with Krebs-Henseleit perfusate (KH) and the luminal side was exposed to conditioned air. SLV, SLD, and CBF were measured with a microscope provided with an electronic micrometer and strobe light. Apparent viscosity and shear elastic modulus were measured with a microcapillary method using mucus collected at the downstream end of the preparation. Control CBF, SLV, and SLD were 11.6 +/- 0.4 (SE) Hz, 91 +/- 8 micron/s, and 33 +/- 5 microns, respectively, at base line and did not change during KH perfusion for 100 min. Perfusion with both acetylcholine and epinephrine (10(-5) to 10(-3) M) produced concentration-dependent increases in mean CBF (maximum increases at 10(-3) M of 16 and 9%, P less than 0.05), whereas only acetylcholine increased mean SLV (+56% at 10(-3) M, P less than 0.05). Perfusion with platelet-activating factor (10(-7) to 10(-5) M) decreased both mean CBF and SLV in a dose-dependent fashion (-6 and -63% at 10(-5) M, P less than 0.05), whereas antigen perfusion (1:60 dilution) increased mean CBF (+10%, P less than 0.05) but decreased SLV (-47%, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Stimulation of ciliary beat frequency by autonomic agonists: in vivo

1988 ◽  
Vol 65 (2) ◽  
pp. 971-981 ◽  
Author(s):  
L. B. Wong ◽  
I. F. Miller ◽  
D. B. Yeates
Keyword(s):  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Base Line ◽  
Muscarinic Cholinergic ◽  
The Mean ◽  
Beta 2 ◽  
Ciliary Beat

beta 2-Adrenergic bronchodilator and muscarinic cholinergic bronchoconstrictor agonists both stimulate ciliary activity in vitro. To test the hypothesis that increases in autonomic activity would result in increases in ciliary beat frequency (CBF) in vivo, a correlation analysis heterodyne laser light-scattering system was developed and validated to measure the stimulating effects of sympathomimetic and parasympathomimetic agonists on tracheal CBF in intact, anesthetized beagles. The mean baseline CBF from 42 studies of 274 measurements in 9 (5 male and 4 female) adult beagles was 6.6 +/- 1.1 Hz. The stimulating effects of a beta 2-adrenergic agonist, fenoterol, and a muscarinic cholinergic agonist, methacholine, on CBF were studied on four and eight beagles, respectively. The studies were randomized and blinded. Aerosolized 10(-5) M fenoterol stimulated the CBF from the base line of 6.8 +/- 2.5 to 32.0 +/- 17.9 Hz in four dogs. Aerosolized methacholine stimulated the CBF from the base line of 5.8 +/- 0.7 to 9.4 +/- 3.0 Hz for 10(-8) M, and to 12.6 +/- 3.1 Hz for 10(-6) M in eight dogs. These are the first data obtained in intact animals that demonstrate CBF in the lower respiratory tract is regulated by autonomic agonists.

Role of ciliary motility in acute allergic mucociliary dysfunction

1982 ◽  
Vol 52 (4) ◽  
pp. 1018-1023 ◽  
Author(s):  
D. R. Maurer ◽  
M. Sielczak ◽  
W. Oliver ◽  
W. M. Abraham ◽  
A. Wanner
Keyword(s):  
Beat Frequency ◽  
Slow Motion ◽  
Ciliary Beat Frequency ◽  
Ciliary Activity ◽  
Base Line ◽  
Conscious Sheep ◽  
Line P ◽  
Chemical Mediators

The purpose of this investigation was to determine if abnormal ciliary function contributes to allergic mucociliary dysfunction. In conscious sheep with Ascaris suum hypersensitivity, ciliated cells were obtained with a cytology brush and tracheal mucous velocity (TMV) was determined before and serially for 2 h following antigen inhalation. The recovered cells (also containing mast cells) were suspended in a chamber, and ciliary activity was viewed microscopically and recorded on videotape for subsequent slow-motion analysis of ciliary beat frequency (CBF). One hour after A. suum challenge mean CBF (+/- SE) showed a slight increase from a base-line value of 630 +/- 16 to 716 +/- 30 beats/min (P less than 0.05) when mean TMV was decreased to 57% of base line (P less than 0.05). After 2 h, both mean CBF and TMV returned toward base line. Since possible in vivo actions of chemical mediators liberated by antigen challenge may have been lost after suspension of the brushed cells, we also assessed the effects of antigen on CBF in vitro. A. suum caused a dose-dependent increase in CBF that was blocked by cromolyn sodium. We conclude that 1) allergic mucociliary dysfunction is not caused by a decrease in CBF and 2) antigen-induced release of chemical mediators increases CBF.

scholarly journals CILIARY ACTIVITY OF BRONCHIAL EPITHELIUM UNDER EXPERIMENTAL COLD EXPOSURE IN VITRO

2017 ◽  
Vol 1 (66) ◽  
pp. 41-49
Author(s):  
Андрей Одиреев ◽  
Andrey Odireev ◽  
Ксения Килимченко ◽  
Kseniya Kilimchenko ◽  
Николай Безруков ◽  
...  
Keyword(s):  
Cold Stress ◽  
Cold Exposure ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Solution Temperature ◽  
Ciliary Activity ◽  
Physiological Level ◽  
Functional Disturbance ◽  
Physiological Conditions

To date, the role of dysfunction of the bronchial ciliated epithelium (BCE) in the formation of mucociliary disorders during cold exposure has not been fully established. The aim of the study was to investigate the nature and severity of changes in the ciliary motility of the BCE under the influence of a cold stimulus in vitro. Ten volunteers with asthma underwent a bronchoscopy with biopsy of the lobar bronchus mucosa. The biopsy specimens were placed in a Hank’s balanced salt solution (HBSS) on a slide, located on the thermal stage, used to simulate the effect of various temperature on the BCE. Ciliary beat frequency (CBF, Hz) was recorded using a microscope, high-sensitivity digital camera and computer with specially developed software. The initial recording of CBF was performed at 24ºC, after which the temperature of HBSS was gradually increased to physiological level and repeated recording was made at the control points (28ºC and 36ºC). Then, the temperature of the solution was lowered and CBF was registered again at 28ºC, 21ºC and 17ºC. The initial CBF of the BCE varied from 6.74 to 3.77 Hz with average of 5.25±1.48 Hz (M±m). There was a statistically significant increase in CBF when the solution was heated: at 28ºC it was 6.33±1.36 Hz (p=0.004) and remained at this level up to 36ºC – 7.14±1.33 Hz (p=0.002). A gradual decrease in the solution temperature produced a reduction in CBF in comparison with the physiological conditions: 28ºC – 6.12±1.31 Hz (p=0.121), 21ºC – 5.27±1.32 Hz (p=0.001) and 17ºC – 3.95±1.18 Hz (p=0.0001). By the end of the experiment, CBF of the BCE decreased almost 2-fold in comparison with the physiological conditions. A mathematical model that characterizes the behavior of BCE cilia under cold stress was developed. Thus, the results of the study demonstrate a marked decrease in the motor activity of the BCE cilia under cold stress, which underlines a significant contribution of the functional disturbance of BCE to the pathophysiological mechanisms of hypersecretory disorders induced by inhalation of cold air.

scholarly journals Combination therapy with cystic fibrosis transmembrane conductance regulator modulators augment the airway functional microanatomy

2016 ◽  
Vol 310 (10) ◽  
pp. L928-L939 ◽  
Author(s):  
Susan E. Birket ◽  
Kengyeh K. Chu ◽  
Grace H. Houser ◽  
Linbo Liu ◽  
Courtney M. Fernandez ◽  
...  
Keyword(s):  
Ion Transport ◽  
Functional Imaging ◽  
Clinical Benefit ◽  
Effective Viscosity ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Clinical Results ◽  
Mucociliary Transport ◽  
Sweat Chloride

Recently approved therapies that modulate CFTR function have shown significant clinical benefit, but recent investigations regarding their molecular mechanism when used in combination have not been consistent with clinical results. We employed micro-optical coherence tomography as a novel means to assess the mechanism of action of CFTR modulators, focusing on the effects on mucociliary clearance. Primary human airway monolayers from patients with a G551D mutation responded to ivacaftor treatment with increased ion transport, airway surface liquid depth, ciliary beat frequency, and mucociliary transport rate, in addition to decreased effective viscosity of the mucus layer, a unique mechanism established by our findings. These endpoints are consistent with the benefit observed in G551D patients treated with ivacaftor, and identify a novel mechanism involving mucus viscosity. In monolayers derived from F508del patients, the situation is more complicated, compounded by disparate effects on CFTR expression and function. However, by combining ion transport measurements with functional imaging, we establish a crucial link between in vitro data and clinical benefit, a finding not explained by ion transport studies alone. We establish that F508del cells exhibit increased mucociliary transport and decreased mucus effective viscosity, but only when ivacaftor is added to the regimen. We further show that improvement in the functional microanatomy in vitro corresponds with lung function benefit observed in the clinical trials, whereas ion transport in vitro corresponds to changes in sweat chloride. Functional imaging reveals insights into clinical efficacy and CFTR biology that significantly impact our understanding of novel therapies.

Modulation of Ciliary Beat Frequency in the Upper Airway by Prostaglandins

1996 ◽  
Vol 10 (4) ◽  
pp. 229-234 ◽  
Author(s):  
Rodney J. Schlosser ◽  
Ramon A. Franco ◽  
Judith M. Czaja ◽  
Thomas V. McCaffrey
Keyword(s):  
Culture Media ◽  
Beat Frequency ◽  
Upper Airway ◽  
Ciliary Beat Frequency ◽  
Prostaglandin E ◽  
Ciliary Activity ◽  
Maximum Increase ◽  
Endogenous Prostaglandins ◽  
Ciliary Beat

Prostaglandins are ubiquitous within tissues and participate in several local regulatory functions. Previous reports have presented conflicting evidence on their role in ciliary activity in the upper airway. We studied the in vitro effects of prostaglandin E2 (PGE2) and a prostaglandin I2 analog (Iloprost) on the ciliary beat frequency (CBF) of human adenoid explants and rabbit sinus mucosa. Tissue explants were maintained in culture at 35°C. PGE2 or Iloprost was added to the culture media at concentrations of 10–6, to 10–12 M. CBF was determined using phase contrast microscopy and microphotometry. PGE2 increased CBF with a maximum increase of 54.6% in rabbit mucosa (P < 0.01) and 20.0% in human adenoid (P < 0.009). Iloprost increased CBF with a maximum increase of 35.2% in rabbit mucosa (P < 0.02) and 16.4% in human adenoid (P < 0.03). Induced production of endogenous prostaglandins was determined by treating specimens with a cyclooxygenase inhibitor (diclofenac) prior to addition of PGE2 or Iloprost. Diclofenac had no effect on the ciliostimulatory effects of PGE2 (P > 0.36), however it significantly decreased the ciliostimulatory effect of Iloprost (P < 0.037). Both PGE2 and Iloprost stimulate ciliary motility, yet the action of PGE2 is direct, whereas Iloprost stimulates cilia by release of endogenous prostaglandins in both the human and rabbit.

scholarly journals Epithelial Cell Culture from Human Adenoids: A Functional Study Model for Ciliated and Secretory Cells

2013 ◽  
Vol 2013 ◽  
pp. 1-13 ◽  
Author(s):  
Claudia González ◽  
Marisol Espinosa ◽  
María Trinidad Sánchez ◽  
Karla Droguett ◽  
Mariana Ríos ◽  
...  
Keyword(s):  
Experimental Model ◽  
Defense Mechanism ◽  
Beat Frequency ◽  
Ciliated Cell ◽  
Functional Study ◽  
Secretory Cells ◽  
Ciliary Activity ◽  
Control Mechanisms ◽  
Mucociliary Transport

Background. Mucociliary transport (MCT) is a defense mechanism of the airway. To study the underlying mechanisms of MCT, we have both developed an experimental model of cultures, from human adenoid tissue of ciliated and secretory cells, and characterized the response to local chemical signals that control ciliary activity and the secretion of respiratory mucinsin vitro.Materials and Methods. In ciliated cell cultures, ciliary beat frequency (CBF) and intracellular Ca2+levels were measured in response to ATP, UTP, and adenosine. In secretory cultures, mucin synthesis and secretion were identified by using immunodetection. Mucin content was taken from conditioned medium and analyzed in the presence or absence of UTP.Results. Enriched ciliated cell monolayers and secretory cells were obtained. Ciliated cells showed a basal CBF of 10.7 Hz that increased significantly after exposure to ATP, UTP, or adenosine. Mature secretory cells showed active secretion of granules containing different glycoproteins, including MUC5AC.Conclusion. Culture of ciliated and secretory cells grown from adenoid epithelium is a reproducible and feasible experimental model, in which it is possible to observe ciliary and secretory activities, with a potential use as a model to understand mucociliary transport control mechanisms.

Muscarinic Ciliostimulation Requires Endogenous Prostaglandin Production

1998 ◽  
Vol 12 (3) ◽  
pp. 203-208 ◽  
Author(s):  
Scott M. Gayner ◽  
Thomas V. McCaffrey
Keyword(s):  
Phospholipase A2 ◽  
Phospholipase C ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Pge2 Production ◽  
Ciliary Activity ◽  
Frequency Increase ◽  
Prostaglandin Production ◽  
Ciliary Beat

Prostaglandin E2 (PGE2) is a known modulator in upper airway ciliary activity and may be involved in the transduction of the muscarinic acetylcholine receptor signal. We studied the in vitro effects of muscarinic ciliostimulation on ciliary beat frequency (CBF) and PGE2 in human adenoid explants to determine whether PGE2 production is an essential step in the signal transduction mechanism. Methacholine applied to adenoid explants significantly increased ciliary beat frequency. This effect was blocked by the application of diclofenac, a cyclooxygenase inhibitor. Using radioimmunoassay, PGE2 production was measured during ciliostimulation with methacholine. Methacholine produced a significant increase in production in PGE2 during ciliostimulation. The roles of phospholipase C and phospholipase A2 in prostaglandin production were investigated by inhibiting these enzymes. D609, a phospholipase C inhibitor, significantly inhibited ciliary beat frequency increase and PGE2 production during methacholine stimulation. However, PACOCF3, a phospholipase A2 inhibitor, did not block ciliary beat frequency increase or PGE2 production in response to methacholine. These data show that phospholipase C is required for PGE2 production and ciliostimulation.

Stimulation of tracheal ciliary beat frequency by localized tissue incision

1990 ◽  
Vol 68 (1) ◽  
pp. 411-416 ◽  
Author(s):  
L. B. Wong ◽  
D. B. Yeates
Keyword(s):  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Neck Surgery ◽  
Ciliary Activity ◽  
Base Line ◽  
Arterial Blood ◽  
Tracheal Pressure ◽  
The Right ◽  
Tissue Trauma ◽  
Ciliary Beat

Since measurements of basal ciliary beat frequency (CBF) were significantly lower in our intact canine experiments than reports of ciliary activity in rabbits involving surgical intervention, we hypothesized that local tissue trauma stimulates CBF. The effects of minor neck surgery on tracheal CBF in eight barbiturate-anesthetized eucapnically ventilated beagles were investigated. Each dog underwent two studies. Measurements of CBF were made at 1-min intervals on the right lateral midtracheal surface by means of heterodyne mode correlation analysis laser light scattering. In the control study, CBF was measured in each dog for at least 160 min. In the incision study, base-line CBF was measured for at least 40 min. The overlying sternohyoidus muscles were then separated, and a longitudinal 2- to 3-cm incision was made in the trachea caudally from the fourth to the fifth cartilage ring. CBF was measured at least 5 cm distally from the site of tracheal injury for an additional 120 min. Electrocardiogram, rectal temperature, tracheal pressure, exhaled CO2, and arterial blood pressure, PO2, PCO2, and pH remained stable throughout both studies. The mean base-line CBF was 4.7 +/- 0.4 Hz. It increased to 19.5 +/- 2.9 Hz (P less than 0.0001) 100 min after the incision and remained elevated until the end of the study period (P less than 0.0001). The mechanism(s) causing this stimulation may also be responsible for the high “basal” CBF observed in other studies.(ABSTRACT TRUNCATED AT 250 WORDS)

The Effects of Sinus Bacteria on Human Ciliated Nasal Epithelium in Vitro

1988 ◽  
Vol 98 (4) ◽  
pp. 299-304 ◽  
Author(s):  
Jonathan L. Ferguson ◽  
Thomas V. McCaffrey ◽  
Eugene B. Kern ◽  
William J. Martin
Keyword(s):  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Nasal Epithelium ◽  
Photometric Method ◽  
Ciliary Activity ◽  
Human Beings ◽  
Minute Period ◽  
Ciliary Motion ◽  
Cell Free Filtrate

The mechanisms by which bacteria colonize and damage ciliated epithelium are important in understanding the pathophysiology of rhinitis, sinusitis, and otitis. Bacteria that have the ability to impair mucociliary clearance would be at an advantage in establishing infection of ciliated surfaces. This study investigates the effect of Hemophilus Influenzae, Streptococcus pneumoniae, Brahmanella catarralis, and Staphylococcus epidermidis on the ciliary activity of normal ciliated nasal epithelium in human beings. Ciliary activity of the nasal epithelium in the presence of each pathogen was assessed for more than 240 minutes with a photometric method of ciliary beat frequency (CBF) measurement. H. influenzae exerted significant effects on ciliary activity, with a 46% decrease in the CBF by 4 hours (with bacteria-containing broth) and a 32% decrease with bacteria-free filtrate. S. epidermidis decreased CBF by 44% with the bacterial broth. A sterile cell-free filtrate had no significant effect. S. pneumoniae and B. catarralis had no significant effect on CBF within a 240-minute period. H. influenzae and S. epidermidis disrupted normal synchronous ciliary motion, causing adjacent cilia to beat at different rates.

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