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S. R. Martins, M. Stastna, J. Castro-Combs, G. Noguera, C. Y. Park, J. V. Eyk, A. Behrens, P. J. McDonnell; Proteomics of Eye - Aqueous Humor Protective Mechanisms Against Bacterial Invasion After Cataract Surgery. Invest. Ophthalmol. Vis. Sci. 2007;48(13):3800.
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Clear cornea cataract surgery has been associated with a higher incidence of infectious endophthalmitis. Classic sutured limbal incision is associated with more inflammation, which may be protective against infection. The purpose of this study is to evaluate the presence of antibacterial proteins in the aqueous humor after using these two incision types in an animal model.
Samples of aqueous humor were taken from 45 New Zealand white rabbits. Animals were divided in 3 groups: no surgery (NS) (n=15), clear cornea incision (CC) of 2.0 mm width (n=15), and limbal incision (LI) of 11 mm width (n=15). CC wounds did not receive any sutures, and LI wounds received 5 interrupted nylon 10-0 sutures. Aqueous samples were taken via paracentesis at 30 min, 1 h, 12 h, 24 h and 48 h after completion of the procedure. Techniques of proteomics analysis (1-D gel electrophoresis (1-DE), 2-D gel electrophoresis (2-DE), reversed phase liquid chromatography (RPLC) and protein identification) were carried out.
Selected methods showed a good reproducibility either of the same sample or of multiple rabbit samples. A total of 92 proteins were identified in the NSG, some of which have antibacterial activity. 2DE analysis of CC and LI showed a dramatic increase in protein content within 30 minutes of surgery, with a peak at 30 min and slow decline to baseline over 48 hrs. The LI displayed ~3 fold greater increase in protein concentration as compared to CC (determined by BCA protein assay), which was clearly seen in the 2DE. There was a large number of unique proteins appearing in the aqueous. Comparison between surgery methods even at baseline and 48 hours show unique set of response.
The presence of proteins with known antibacterial activity in the aqueous humor of the rabbit may explain a status of relative resistance to infection after intraocular surgery. Increased amount of these proteins in the LI vs. CC in this animal model may correlate well with the lower incidence of endophthalmitis in humans in the classic old method of cataract surgery. Further characterization of these proteins in the human eye is required to confirm our hypothesis.
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