ciliaFA: a research tool for automated, high-throughput measurement of ciliary beat frequency using freely available software

Smith, Claire M.; Djakow, Jana; Free, Robert C; Djakow, Petr; Lonnen, Rana; Williams, Gwyneth; Pohunek, Petr; Hirst, Robert A.; Easton, Andrew J.; Andrew, Peter W.; O’Callaghan, Christopher

doi:10.1186/2046-2530-1-14

Cited by 83 publications

(87 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Automated software CiliaFA was used, which runs on opensource software (ImageJ) (Smith et al, 2012). The software extracts the pixel intensities of the region of interest (ROI) over time from previously recorded video.…”

Section: Organotypic Tissue Culture Modelsmentioning

confidence: 99%

3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product

Iskandar

Mathis

Martin

et al. 2017

ALTEX

View full text Add to dashboard Cite

ratory animals -to reduce the number of animals, to refine the design of procedures such that pain and distress are minimized, and to replace animal models with alternative methods and lower organisms when possible (Doke and Dhawale, 2015). In this regard, the capability to preserve human respiratory cells in culture provides a tool not only to generate mechanistic data, but also to improve the predicted effects of exposure hazard through inhalation in humans. An emphasis on assessment studies using human-derived in vitro culture systems will reduce the problems that are inherent to interspecies translatability (Manuppello and Sullivan, 2015).In vitro toxicology approaches have evolved from focusing on the molecular changes within a cell to understanding the toxicity-related mechanisms (cell-cell interactions) in models IntroductionMechanistic investigation of the impact of exposure on the lower airway remains challenging because access to lung tissues is limited. Although biopsy samples can be obtained from patients/donors for mechanistic studies of exposure-induced injury, a biopsy procedure is invasive and there are related ethical concerns (Peppercorn, 2013). Alternatively, animal studies allow the collection of biological samples from various animal models of human diseases. However, biological responses can vary among animal species, and findings obtained from animal studies may not apply to humans. Alternatives to animal testing have been proposed to overcome these drawbacks, including the 3Rs strategy -reduction, refinement, and replacement of labo- Research SummaryIn vitro toxicology approaches have evolved from a focus on molecular changes within a cell to understanding of toxicity-related mechanisms in systems that can mimic the in vivo environment. The recent development of three dimensional (3-D) organotypic nasal epithelial culture models offers a physiologically robust system for studying the effects of exposure through inhalation. Exposure to cigarette smoke (CS) is associated with nasal inflammation; thus, the nasal epithelium is relevant for evaluating the pathophysiological impact of CS exposure. The present study investigated further the application of in vitro human 3-D nasal epithelial culture models for toxicological assessment of inhalation exposure. Aligned with 3Rs strategy, this study aimed to explore the relevance of a human 3-D nasal culture model to assess the toxicological impact of aerosols generated from a candidate modified risk tobacco product (cMRTP), the Tobacco Heating System (THS) 2.2, as compared with smoke generated from reference cigarette 3R4F. A series of experimental repetitions, where multiple concentrations of THS2.2 aerosol and 3R4F smoke were applied, were conducted to obtain reproducible measurements to understand the cellular/molecular changes that occur following exposure. In agreement with "Toxicity Testing in the 21 st Century -a Vision and a Strategy", this study implemented a systems toxicology approach and found that for all tested concentrations the impa...

show abstract

Section: Organotypic Tissue Culture Modelsmentioning

confidence: 99%

3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product

Iskandar

Mathis

Martin

et al. 2017

ALTEX

View full text Add to dashboard Cite

show abstract

“…Cell cultures (at the w5 time point only) were visualized using an Axiovert 200 M microscope (Carl Zeiss, Germany) connected to a Sony XCD V60 Firewire camera. A total 256 images of the cultures were recorded per insert, and CBF was then calculated using Cilia FA software as described by Smith et al 61 Each condition was tested in triplicate on four independent occasions (n = 4).…”

mentioning

confidence: 99%

Human Asthmatic Bronchial Cells Are More Susceptible to Subchronic Repeated Exposures of Aerosolized Carbon Nanotubes At Occupationally Relevant Doses Than Healthy Cells

et al. 2017

View full text Add to dashboard Cite

Although acute pulmonary toxicity of carbon nanotubes (CNTs) has been extensively investigated, the knowledge of potential health effects following chronic occupational exposure is currently limited and based only upon in vivo approaches. Our aim was to realistically mimic subchronic inhalation of multiwalled CNTs (MWCNTs) in vitro, using the air−liquid interface cell exposure (ALICE) system for aerosol exposures on reconstituted human bronchial tissue from healthy and asthmatic donors. The reliability and sensitivity of the system were validated using crystalline quartz (DQ12), which elicited an increased (pro-)inflammatory response, as reported in vivo. At the administrated MWCNT doses relevant to human occupational lifetime exposure (10 μg/cm 2 for 5 weeks of repeated exposures/5 days per week) elevated cilia beating frequency (in both epithelial cultures), and mucociliary clearance (in asthmatic cells only) occurred, whereas no cytotoxic reactions or morphological changes were observed. However, chronic MWCNT exposure did induce an evident (pro-)inflammatory and oxidative stress response in both healthy and asthmatic cells. The latter revealed stronger and more durable long-term effects compared to healthy cells, indicating that individuals with asthma may be more susceptible to adverse effects from chronic MWCNT exposure. Our results highlight the power of occupationally relevant subchronic exposures on human in vitro models in nanosafety hazard assessment. KEYWORDS: in vitro primary lung system, reconstituted healthy and asthma tissue, air−liquid interface, multiwalled carbon nanotubes, subchronic repeated exposure, occupational doses, hazard assessment T he high aspect ratio, excellent strength, and good conductivity offered by carbon nanotubes (CNTs) has raised their demand in the global industry market.

show abstract

“…Several groups have attempted to overcome these problems by developing software to automate analysis from the digital images [53][54][55].…”

Section: Page 5 Of 36mentioning

confidence: 99%

“…Although assessment of ciliary function by light microscopy is recognised as a critical diagnostic test used as 'first-line' in many centres [11,17,48,53,54,56] it should be noted that some clinicians question its value and recommend that HVMA should be reserved for research purposes [20]. A limitation of HVMA is the need for specialist equipment, rigorous quality control and experienced technicians with a high through-put of normal and abnormal samples.…”

Section: Page 5 Of 36mentioning

confidence: 99%

Diagnostic Methods in Primary Ciliary Dyskinesia

Lucas

Paff

Goggin

et al. 2016

Paediatric Respiratory Reviews

View full text Add to dashboard Cite

Educational Aims; The reader will be able to; Describe the basis of the various diagnostic tests for PCD  Have an understanding of the strengths and limitations of each test  Understand that there is no 'gold standard' test  Understand why highly specialised centres should analyse samples and interpret results. Page 2 of 36A c c e p t e d M a n u s c r i p t  SummaryDiagnosing primary ciliary dyskinesia is difficult. With no reference standard, a combination of tests is needed; most tests require expensive equipment and specialist scientists. We review the advances in diagnostic testing over the past hundred years, with emphasis on recent advances.We particularly focus on use of high-speed video analysis, transmission electron microscopy, nasal nitric oxide and genetic testing. We discuss the international efforts that are in place to advance the evidence base for diagnostic tests.

show abstract

ciliaFA: a research tool for automated, high-throughput measurement of ciliary beat frequency using freely available software

Cited by 83 publications

References 19 publications

3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product

3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product

Human Asthmatic Bronchial Cells Are More Susceptible to Subchronic Repeated Exposures of Aerosolized Carbon Nanotubes At Occupationally Relevant Doses Than Healthy Cells

Diagnostic Methods in Primary Ciliary Dyskinesia

Contact Info

Product

Resources

About