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Transplantation of Human Amniotic Membrane Influences Matrix Metalloproteinases (MMPs) and TIMPs in a Murine Model with Herpertic Keratitis
1Heiligenhaus A., 1Li H., 1Yang Y., 2Wasmuth S., 1Bauer D., 3Steuhl K. P.,
1St. Franziskus-Hospital, Augenabteilung (Münster)
2Universität-Gesamthochschule Essen, Zentrum für Augenheilkunde (Essen)
3Universität-Gesamthochschule Essen, Zentrum für Augenheilkunde, Abt. für Erkrankungen des vorderen Augenabschnitts (Essen)
Purpose: There is evidence that amniotic membrane transplantation (AMT) improves the ulcerative process of herpes simplex virus type-1 (HSV-1) keratitis in mice. The authors now studied whether AMT influences metalloproteinases (MMPs) and TIMPs in the corneas.
Methods: BALB/c mice were corneally infected with HSV-1. For the experiments, mice were chosen that presented with corneal ulcers by day 14 p.i. Whereas the corneas of mice in group 1 were covered with AMT, tarsorrhaphies were performed in others (group 2). After 2 days, the appearance of corneal ulcers and stromal inflammation was judged clinically. Corneal sections were studied by immunohistochemistry for the expression of MMP-2, -8 and -9 and TIMP-1 and -2. MMP activity in the corneas was investigated by zymography, and the expression of the enzymes was measured by Western blot technique.
Results: At day 14 p.i., the ulcers were highly positive for MMP-2, -8 and -9, and TIMP-1 and TIMP-2, and the active forms of these MMPs and TIMPs were detected. By zymography, the gelatinolytic and caseinolytic activities were found in the corneas. In the mice of group 1, the ulceration and stromal inflammation markedly improved within 2 days. The immunohistochemical and Western blot studies revealed that this was associated with a reduced expression of MMPs. The MMP enzymatic activity was reduced. This was contrasted by the findings in group 2.
Conclusions: Rapid improvement of HSV-1 induced ulcerative keratitis in mice is noted after amniotic membrane transplantation. The findings suggest that this may result at least in part from a reduced expression and activity of MMPs.