Rapid method of quantification of tight‐junction organization using image analysis

Terryn, Christine; Sellami, Mehdi; Fichel, Caroline; Diebold, Marie-Danielle; Gangloff, Sophie C.; Naour, Richard Le; Polette, Myriam; Zahm, Jean‐Marie

doi:10.1002/cyto.a.22239

Cited by 24 publications

(26 citation statements)

References 7 publications

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“…ImageJ, a public-domain Java image processing program that is freely available from the National Institutes of Health (http://rsb.info.nih.gov/ij/), has been shown to be extremely effective at measuring the area of elliptical or irregularly shaped selections. [23][24][25] The program simplifies area calculations by digitally measuring the number of pixels within a selection. As such, it may be easily be applied to the previously mentioned glenoid bone loss quantification schemes for enhancement of accuracy in measuring the area of a best-fit circle, as well as the area of the irregularly shaped bone defect.…”

Section: Discussionmentioning

confidence: 99%

Glenoid Diameter Is an Inaccurate Method for Percent Glenoid Bone Loss Quantification: Analysis and Techniques for Improved Accuracy

Bhatia

Saigal

Frank

et al. 2015

Arthroscopy: The Journal of Arthroscopic & Related Surgery

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Section: Discussionmentioning

confidence: 99%

Glenoid Diameter Is an Inaccurate Method for Percent Glenoid Bone Loss Quantification: Analysis and Techniques for Improved Accuracy

Bhatia

Saigal

Frank

et al. 2015

Arthroscopy: The Journal of Arthroscopic & Related Surgery

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“…Immunofluorescent images were obtained using an Olympus FV-1000 confocal microscope (Olympus Corporation, Tokyo, Japan). The tight junction organization rate (TiJOR) was calculated using an ImageJ software macro (88) to evaluate the damage to tight junction networks. TiJOR (entire image) was calculated by evaluating the entire representative images obtained by confocal microscopy.…”

Section: Methodsmentioning

confidence: 99%

Bordetella pertussis Adenylate Cyclase Toxin Disrupts Functional Integrity of Bronchial Epithelial Layers

et al. 2018

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The airway epithelium restricts the penetration of inhaled pathogens into the underlying tissue and plays a crucial role in the innate immune defense against respiratory infections. The whooping cough agent, , adheres to ciliated cells of the human airway epithelium and subverts its defense functions through the action of secreted toxins and other virulence factors. We examined the impact of infection and of adenylate cyclase toxin-hemolysin (CyaA) action on the functional integrity of human bronchial epithelial cells cultured at the air-liquid interface (ALI). adhesion to the apical surface of polarized pseudostratified VA10 cell layers provoked a disruption of tight junctions and caused a drop in transepithelial electrical resistance (TEER). The reduction of TEER depended on the capacity of the secreted CyaA toxin to elicit cAMP signaling in epithelial cells through its adenylyl cyclase enzyme activity. Both purified CyaA and cAMP-signaling drugs triggered a decrease in the TEER of VA10 cell layers. Toxin-produced cAMP signaling caused actin cytoskeleton rearrangement and induced mucin 5AC production and interleukin-6 (IL-6) secretion, while it inhibited the IL-17A-induced secretion of the IL-8 chemokine and of the antimicrobial peptide beta-defensin 2. These results indicate that CyaA toxin activity compromises the barrier and innate immune functions ofinfected airway epithelia.

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“…The formation intensity and solidity of tight junctions were analysed using image quantification using Fuji 58 based upon an adapted method previously reported by Terryn et al 59 and McNeil et al 60 . In brief, a baseline threshold was determined prior to analysis.…”

Section: Setting Up the Quasi Vivo 600mentioning

confidence: 99%

A dynamic perfusion based blood-brain barrier model for cytotoxicity testing and drug permeation

Elbakary

Badhan

2020

Sci Rep

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the blood-brain barrier (BBB) serves to protect and regulate the cnS microenvironment. the development of an in-vitro mimic of the BBB requires recapitulating the correct phenotype of the invivo BBB, particularly for drug permeation studies. However the majority of widely used BBB models demonstrate low transendothelial electrical resistance (TEER) and poor BBB phenotype. The application of shear stress is known to enhance tight junction formation and hence improve the barrier function. We utilised a high teeR primary porcine brain microvascular endothelial cell (pBMec) culture to assess the impact of shear stress on barrier formation using the Kirkstall QuasiVivo 600 (QV600) multi-chamber perfusion system. the application of shear stress resulted in a reorientation and enhancement of tight junction formation on both coverslip and permeable inserts, in addition to enhancing and maintaining TEER for longer, when compared to static conditions. Furthermore, the functional consequences of this was demonstrated with the reduction in flux of mitoxantrone across PBMEC monolayers. The QV600 perfusion system may service as a viable tool to enhance and maintain the high teeR pBMec system for use in in-vitro BBB models. The blood-brain barrier (BBB) represents an insidious barrier for the delivery of therapeutic agents for a wide range of central nervous system (CNS) disorders. Penetration of the restrictive brain microvascular endothelial cell barrier is often hindered by the presence of a network of intra-cellular tight junction proteins, in addition to a network of membrane localised active transporter proteins and enzymatic metabolism processes. The development and maintenance of an appropriate restrictive in-vitro BBB model is critical when assessing the potential for small molecule transport. Despite the rise in the use of in-vivo models for assessing BBB structure and function, in-vitro models are still widely used and have been developed from a range of species. However, a consensus on the most appropriate cellular system has still not been achieved, particularly in the context of assessing drug permeation and inherent barrier properties. For example, the human immortalised hCMEC/D3 cell, when grown in co-culture with astrocytes, yields low TEER values of approximately 140 Ω.cm 2 1 , and those from primary endothelial cells from rodents yield TEER values of approximately 300 Ω.cm 2 2. Higher TEER values have been obtained with stem cell based systems (iPSC-derived endothelial cells) and neuronal progenitor cells, when exposed to chemical treatment, resulting in values of 3000-4000 Ω.cm 2 3 to promote BBB formation, although these often require specialised and costly methods to culture. Although human brain tissue derived in-vitro BBB models are idealised for BBB studies, the lack of appropriate monolayer formation and reproductively, in-vitro, has led to other cellular models being attractive options. The use of a porcine primary cell culture system (PBMEC) reporting high TEER without the need for co-culture with as...

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Rapid method of quantification of tight‐junction organization using image analysis

Cited by 24 publications

References 7 publications

Glenoid Diameter Is an Inaccurate Method for Percent Glenoid Bone Loss Quantification: Analysis and Techniques for Improved Accuracy

Glenoid Diameter Is an Inaccurate Method for Percent Glenoid Bone Loss Quantification: Analysis and Techniques for Improved Accuracy

Bordetella pertussis Adenylate Cyclase Toxin Disrupts Functional Integrity of Bronchial Epithelial Layers

A dynamic perfusion based blood-brain barrier model for cytotoxicity testing and drug permeation

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