The Scar-in-a-Jar: studying potential anti?brotic compounds from the epigenetic to extracellular level in a single well
MetadataShow full item record
Background and purpose: Fibrosis, a pathological accumulation of collagen in tissues, represents a major global disease burden. Effective characterization of potential antifibrotic drugs has been constrained by poor formation of the extracellular matrix in vitro, due to tardy procollagen processing by collagen C-proteinase/BMP-1, and difficulties in relating this matrix to cell numbers in experimental samples. Experimental approach: The Scar-in-a-Jar model provided, in vitro, the complete biosynthetic cascade of collagen matrix formation including complete conversion of procollagen by C-proteinase/BMP-1, its subsequent extracellular deposition and lysyl oxidase-mediated cross-linking, achieved by applying the biophysical principle of macromolecular ‘crowding’. Collagen matrix deposition, velocity and morphology can be controlled using negatively charged ‘crowders’ in a rapid (2 days) mode or a mixture of neutral ‘crowders’ in an accelerated (6 days) mode. Combined with quantitative optical bioimaging, this novel system allows for in situ assessment of the area of deposited collagen(s) per cell. Key results: Optical evaluation of known and novel antifibrotic compounds effective at the epigenetic, post-transcriptional/translational/secretional level correlated excellently with corresponding biochemical analyses.Focusing on quantitation of deposited collagen, the Scar-in-a-Jar was most effective in assessing novel inhibitors that may have multiple targets, such as microRNA29c, found to be a promising antifibrotic agent. Conclusions and implications: This novel screening system supersedes current in vitro fibroplasia models, as a fast, quantitative and non-destructive technique. This method distinguishes a reduction in collagen I deposition, excluding collagen cross-linking, and allows full evaluation of inhibitors of C-proteinase/BMP-1 and other matrix metalloproteinases.
Showing items related by title, author, creator and subject.
High-resolution study of the 3D collagen fibrillary matrix of Achilles tendons without tissue labelling and dehydratingWu, J.; Swift, B.; Becker, Thomas; Squelch, Andrew; Wang, A.; Zheng, Y.; Zhao, X.; Xu, J.; Xue, W.; Zheng, M.; Lloyd, D.; Kirk, Brett (2017)Knowledge of the collagen structure of an Achilles tendon is critical to comprehend the physiology, biomechanics, homeostasis and remodelling of the tissue. Despite intensive studies, there are still uncertainties regarding ...
High-resolution study of the 3D collagen fibrillary matrix of Achilles tendons without tissue labelling and dehydratingWu, Jian-Ping; Swift, B.; Becker, Thomas; Squelch, Andrew; Wang, A.; Zheng, Y.; Zhao, X.; Xu, J.; Xue, W.; Zheng, M.; Lloyd, D.; Kirk, Brett (2017)Â© 2017 The Authors Journal of Microscopy Â© 2017 Royal Microscopical Society Knowledge of the collagen structure of an Achilles tendon is critical to comprehend the physiology, biomechanics, homeostasis and remodelling ...
Wang, Z.; Chen, C.; Finger, S.; Kwajah, M.; Jung, M.; Schwarz, H.; Swanson, N.; Lareu, Ricardo; Raghunath, M. (2009)Pulmonary fibrosis represents a fatal stage of interstitial lung diseases of known and idiopathic aetiology. No effective therapy is currently available. Based on an indication-discovery approach we present novel in vitro ...