Evaluation of regenerated tracheal cilia function on a collagen-conjugated scaffold in a canine model

Abstract OBJECTIVES It is unclear whether the movement and function of the regenerated cilia on collagen-conjugated artificial trachea are the same as those of normal cilia. This study assessed the ciliary beat frequency (CBF) and ciliary transport functions (CTFs) of regenerated cilia in a canine model. METHODS A tracheal defect introduced into the anterior portion of the cervical trachea of an adult beagle dog was covered with a collagen-conjugated prosthesis. Two months later, the trachea was harvested along the long axis, both from normal and regenerated regions. The cilia were stained with isothiocyanate-conjugated wheat germ agglutinin, and their movement was monitored with a high-speed camera to analyse CBF and CTF. Four samples each were obtained from the regenerated and normal regions for CBF analysis and 7 samples each were obtained for CTF analysis. RESULTS The wheat germ agglutinin-stained cells showed well-regulated beats in both the regenerated and normal regions of the trachea. Mean CBF in the regenerated and normal regions did not differ significantly (7.11 ± 0.41 vs 7.14 ± 1.09 Hz; P = 981). By contrast, CTF was significantly lower in the regenerated region than in the normal region (30.0 ± 6.62 vs 7.43 ± 0.58 μm/s; P = 0.005). CONCLUSIONS Mean CBF in the regenerated and normal regions did not differ significantly at 2 months. The CTF in the regenerated region recovered partially but remained lower than those in the normal region. Methods are needed to improve the CTF of regenerated cilia.

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Betadine has a ciliotoxic effect on ciliated human respiratory cells

The Journal of Laryngology & Otology ◽

10.1017/s0022215114002746 ◽

2014 ◽

Vol 129 (S1) ◽

pp. S45-S50 ◽

Cited By ~ 20

Author(s):

J H Kim ◽

J Rimmer ◽

N Mrad ◽

S Ahmadzada ◽

R J Harvey

Keyword(s):

Epithelial Cells ◽

High Speed ◽

Imaging System ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Positive Control ◽

Clinical Dose ◽

Significant Difference ◽

Sinonasal Mucosa ◽

Ciliary Beat

AbstractObjective:This study investigated the effect of Betadine on ciliated human respiratory epithelial cells.Methods:Epithelial cells from human sinonasal mucosa were cultured at the air–liquid interface. The cultures were tested with Hanks' balanced salt solution containing 10 mM HEPES (control), 100 µM ATP (positive control), 5 per cent Betadine or 10 per cent Betadine (clinical dose). Ciliary beat frequency was analysed using a high-speed camera on a computer imaging system.Results:Undiluted 10 per cent Betadine (n = 6) decreased the proportion of actively beating cilia over 1 minute (p < 0.01). Ciliary beat frequency decreased from 11.15 ± 4.64 Hz to no detectable activity. The result was similar with 5 per cent Betadine (n = 7), with no significant difference compared with the 10 per cent solution findings.Conclusion:Betadine, at either 5 and 10 per cent, was ciliotoxic. Caution should be applied to the use of topical Betadine solution on the respiratory mucosal surface.

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Quantification of Ciliary Beat Frequency in Sinonasal Epithelial Cells Using Differential Interference Contrast Microscopy and High-Speed Digital Video Imaging

American Journal of Rhinology ◽

10.1177/194589240602000123 ◽

2006 ◽

Vol 20 (1) ◽

pp. 124-127 ◽

Cited By ~ 32

Author(s):

Ioana Schipor ◽

James N. Palmer ◽

Akiva S. Cohen ◽

Noam A. Cohen

Keyword(s):

Epithelial Cells ◽

High Speed ◽

Digital Video ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Differential Interference Contrast ◽

Differential Interference Contrast Microscopy ◽

Video Imaging ◽

Contrast Microscopy ◽

Interference Contrast Microscopy

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Deep phenotyping, including quantitative ciliary beating parameters, and extensive genotyping in primary ciliary dyskinesia

Journal of Medical Genetics ◽

10.1136/jmedgenet-2019-106424 ◽

2019 ◽

Vol 57 (4) ◽

pp. 237-244 ◽

Cited By ~ 2

Author(s):

Sylvain Blanchon ◽

Marie Legendre ◽

Mathieu Bottier ◽

Aline Tamalet ◽

Guy Montantin ◽

...

Keyword(s):

Primary Ciliary Dyskinesia ◽

High Speed ◽

Genetic Disorder ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Central Complex ◽

Ciliary Beating ◽

Recovery Stroke ◽

Biallelic Mutations ◽

Ciliary Dyskinesia

BackgroundPrimary ciliary dyskinesia (PCD) is a rare genetic disorder resulting in abnormal ciliary motility/structure, extremely heterogeneous at genetic and ultrastructural levels. We aimed, in light of extensive genotyping, to identify specific and quantitative ciliary beating anomalies, according to the ultrastructural phenotype.MethodsWe prospectively included 75 patients with PCD exhibiting the main five ultrastructural phenotypes (n=15/group), screened all corresponding PCD genes and measured quantitative beating parameters by high-speed video-microscopy (HSV).ResultsSixty-eight (91%) patients carried biallelic mutations. Combined outer/inner dynein arms (ODA/IDA) defect induces total ciliary immotility, regardless of the gene involved. ODA defect induces a residual beating with dramatically low ciliary beat frequency (CBF) related to increased recovery stroke and pause durations, especially in case of DNAI1 mutations. IDA defect with microtubular disorganisation induces a low percentage of beating cilia with decreased beating angle and, in case of CCDC39 mutations, a relatively conserved mean CBF with a high maximal CBF. Central complex defect induces nearly normal beating parameters, regardless of the gene involved, and a gyrating motion in a minority of ciliated edges, especially in case of RSPH1 mutations. PCD with normal ultrastructure exhibits heterogeneous HSV values, but mostly an increased CBF with an extremely high maximal CBF.ConclusionQuantitative HSV analysis in PCD objectives beating anomalies associated with specific ciliary ultrastructures and genotypes. It represents a promising approach to guide the molecular analyses towards the best candidate gene(s) to be analysed or to assess the pathogenicity of the numerous sequence variants identified by next-generation-sequencing.

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Bovine sperm-oviduct interactions are characterized by specific sperm behaviour, ultrastructure and tubal reactions which are impacted by sex sorting

Scientific Reports ◽

10.1038/s41598-020-73592-1 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Miguel Camara Pirez ◽

Heather Steele ◽

Sven Reese ◽

Sabine Kölle

Keyword(s):

Beat Frequency ◽

Ciliary Beat Frequency ◽

Sperm Binding ◽

Tail Movement ◽

Bovine Spermatozoa ◽

Sperm Reservoir ◽

And Function ◽

The Impact

Abstract To date sperm-oviduct interactions have largely been investigated under in vitro conditions. Therefore we set out to characterize the behaviour of bovine spermatozoa within the sperm reservoir under near in vivo conditions and in real-time using a novel live cell imaging technology and a newly established fluorescent sperm binding assay. Sperm structure and tubal reactions after sperm binding were analysed using scanning and transmission electron microscopy and histochemistry. As a model to specify the impact of stress on sperm-oviduct interactions, frozen-thawed conventional and sex-sorted spermatozoa from the same bulls (n = 7) were co-incubated with oviducts obtained from cows immediately after slaughter. Our studies revealed that within the oviductal sperm reservoir agile (bound at a tangential angle of about 30°, actively beating undulating tail), lagging (bound at a lower angle, reduced tail movement), immotile (absence of tail movement) and hyperactivated (whip-like movement of tail) spermatozoa occur, the prevalence of which changes in a time-dependent pattern. After formation of the sperm reservoir, tubal ciliary beat frequency is significantly increased (p = 0.022) and the epithelial cells show increased activity of endoplasmic reticula. After sex sorting, spermatozoa occasionally display abnormal movement patterns characterized by a 360° rotating head and tail. Sperm binding in the oviduct is significantly reduced (p = 0.008) following sexing. Sex-sorted spermatozoa reveal deformations in the head, sharp bends in the tail and a significantly increased prevalence of damaged mitochondria (p < 0.001). Our results imply that the oviductal cells specifically react to the binding of spermatozoa, maintaining sperm survival within the tubal reservoir. The sex-sorting process, which is associated with mechanical, chemical and time stress, impacts sperm binding to the oviduct and mitochondrial integrity affecting sperm motility and function.

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Altered Sinonasal Ciliary Dynamics in Chronic Rhinosinusitis

American Journal of Rhinology ◽

10.2500/ajr.2006.20.2870 ◽

2006 ◽

Vol 20 (3) ◽

pp. 325-329 ◽

Cited By ~ 62

Author(s):

Bei Chen ◽

Jeffrey Shaari ◽

Steven Eau Claire ◽

James N. Palmer ◽

Alexander G. Chiu ◽

...

Keyword(s):

Chronic Rhinosinusitis ◽

Adenosine Triphosphate ◽

High Speed ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Ciliary Activity ◽

Differential Interference Contrast Microscopy ◽

Dynamic Regulation ◽

Mucus Clearance ◽

Persistent Inflammation

Background Although multiple etiologies contribute to the development of rhinosinusitis, a common pathophysiological sequelae is ineffective sinonasal mucociliary clearance, resulting in stasis of sinonasal secretions, with subsequent infection, and persistent inflammation. The respiratory cilia beat continually at a basal rate, while during times of stress, such as exercise or infection, ciliary beat frequency (CBF) increases, accelerating mucus clearance. Previous investigations have led to conflicting results with some authors reporting decreased CBF while others have found normal values of CBF in patients with chronic rhinosinusitis (CRS). Additionally, these studies have only analyzed basal CBF. The goal of this study was to compare the basal as well as the stimulated sinonasal CBF in patients with CRS versus controls. Methods A dual temperature controlled perfusion chamber, differential interference contrast microscopy, and high-speed digital video were used to analyze both basal and adenosine triphosphate (100 μM)–stimulated CBF in human sinonasal mucosal explants. Results Although no difference in basal CBF was detected between control and CRS patients, a marked difference in stimulated CBF was noted. Exogenously applied adenosine triphosphate resulted in a 50–70% increase of CBF in control tissue with a minimally observed CBF increase in explants from CRS patients. Conclusion Dynamic regulation of respiratory ciliary activity is critical for the respiratory epithelium to adapt to varying environmental situations. Thus, diminished or absent adaptation could predispose the sinonasal cavity to accumulation of inhaled infectious and noxious particulate matter resulting in infection/inflammation. Our findings suggest that CRS patients have decreased sinonasal ciliary adaptation to environmental stimuli.

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Effect of Oxymetazoline on Healthy Human Nasal Ciliary Beat Frequency Measured with High-Speed Digital Microscopy and Mucociliary Transport Time

Annals of Otology Rhinology & Laryngology ◽

10.1177/000348940811700211 ◽

2008 ◽

Vol 117 (2) ◽

pp. 127-133 ◽

Cited By ~ 8

Author(s):

Luo Zhang ◽

Demin Han ◽

Xiaohong Song ◽

Kuiji Wang ◽

Hong Wang

Keyword(s):

High Speed ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Mucociliary Transport ◽

Digital Microscopy ◽

Transport Time ◽

Healthy Human ◽

Ciliary Beat

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Intracellular Cl− Regulation of Ciliary Beating in Ciliated Human Nasal Epithelial Cells: Frequency and Distance of Ciliary Beating Observed by High-Speed Video Microscopy

International Journal of Molecular Sciences ◽

10.3390/ijms21114052 ◽

2020 ◽

Vol 21 (11) ◽

pp. 4052

Author(s):

Makoto Yasuda ◽

Taka-aki Inui ◽

Shigeru Hirano ◽

Shinji Asano ◽

Tomonori Okazaki ◽

...

Keyword(s):

Epithelial Cells ◽

High Speed ◽

Beat Frequency ◽

Ciliary Beat Frequency ◽

Video Microscopy ◽

Nasal Epithelial Cells ◽

Ciliary Beating ◽

High Speed Video ◽

Human Nasal Epithelial Cells ◽

Ciliary Beat

Small inhaled particles, which are entrapped by the mucous layer that is maintained by mucous secretion via mucin exocytosis and fluid secretion, are removed from the nasal cavity by beating cilia. The functional activities of beating cilia are assessed by their frequency and the amplitude. Nasal ciliary beating is controlled by intracellular ions (Ca2+, H+ and Cl−), and is enhanced by a decreased concentration of intracellular Cl− ([Cl−]i) in ciliated human nasal epithelial cells (cHNECs) in primary culture, which increases the ciliary beat amplitude. A novel method to measure both ciliary beat frequency (CBF) and ciliary beat distance (CBD, an index of ciliary beat amplitude) in cHNECs has been developed using high-speed video microscopy, which revealed that a decrease in [Cl−]i increased CBD, but not CBF, and an increase in [Cl−]i decreased both CBD and CBF. Thus, [Cl−]i inhibits ciliary beating in cHNECs, suggesting that axonemal structures controlling CBD and CBF may have Cl− sensors and be regulated by [Cl−]i. These observations indicate that the activation of Cl− secretion stimulates ciliary beating (increased CBD) mediated via a decrease in [Cl−]i in cHNECs. Thus, [Cl−]i is critical for controlling ciliary beating in cHNECs. This review introduces the concept of Cl− regulation of ciliary beating in cHNECs.

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