Fibrosis is involve in wound healing mechanism and represents a major global disease burden. The formation of insoluble pericellular collagen matrix is linked to fibrotic processes and could be assess in vitro for screening products efficacy. To improve collagen deposition in vitro, we use charged macromolecules to increase collagen nucleation lead to a granular deposition on human primary fibroblast. This approach is based on the creation of the excluded volume effect (Lareu et al, 2007).
Figure 1 : Enhanced extra cellular matrix deposition in vitro.
In vitro imaging of pericellular matrix required a multiplex protein labbeling for fluorescence high content imaging using our Micro XLS ImageXpress (Molecular Devices). After 5 days of exposure to L-Ascorbic Acid 2-Phosphate Sesquimagnesium (ASC), pericellular matrix was labbeled and quantified.
The amount of Pro Collagen 1, Collagen 1 and Fibronectin was quantified per cells and compared to control condition (CTRL).
Thought this model pro and anti fibrotic compounds can be evaluated during several days.
Figure 2 : Acide ascorbic reduce intracellular pro collagen 1 and enhance collagen 1 deposition.
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