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Demodex-associated Bacillus proteins induce an aberrant wound healing response in a corneal epithelial cell line (hTCEpi).

Invest Ophthalmol Vis Sci. 2012 Apr 24;

Authors: O'Reilly N, Gallagher C, Katikireddy K, Clynes M, O'Sullivan F, Kavanagh K

Abstract
Purpose: The aim of the work presented here was to establish the response of a corneal epithelial cell line (hTCEpi) to protein extracted from a bacterium (Bacillus oleronius) previously isolated from a Demodex mite from a rosacea patient.Methods: The response of the corneal epithelial cell line to Bacillus proteins was measured in terms of alterations in cell migration and invasiveness. Changes in the expression of metalloproteinase genes and proteins were also assessed. Results: The results indicated increased cell migration (14.5 fold, p = 0.001) as measured using 8 µm PET inserts (BD Falcon) in a transwell assay and invasiveness (1.7 fold, p = 0.003) as measured using 8 µm Matrigel (BD Biocoat) invasion inserts in a 24 well plate assay format, following exposure to the Bacillus proteins. Cells exposed to the Bacillus protein showed a dose dependent increased in expression of genes coding for matrix metalloprotease-3 (MMP-3) (61 fold) and matrix metalloprotease-9 (MPP-9) (301 fold). This dose dependent increase in gene expression was also reflected in elevated levels of MMP-9 protein (1.34 fold, p = 0.033) and increased matrix metalloprotease activity (1.96 fold, p = 0.043) being present in the culture supernatent. Cells also displayed reduced levels of β-integrin (1.25 fold, p = 0.01), indicative of increased motility, and elevated levels of vinculin (2.7 fold, p = 0.0009), suggesting altered motility. Conclusion: The results indicate that exposure of corneal epithelial cells to Bacillus proteins results in an aberrant wound healing response as visualized using a scratch wound assay.

PMID: 22531699 [PubMed - as supplied by publisher]

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?tmpl=NoSidebarfile&db=PubMed&cmd=Retrieve&list_uids=22531699&dopt=Abstract = URL to article

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