Background: The eye is frequently involved in patients with scleroderma. Most often, this involvement consists of scleroderma of the eyelids, resulting in tightness of the lids and blepharophimosis. Also, keratoconjunctivitis sicca (KCS) with a Sjogren's-like picture has been described.
Objectives: This study analyzes and compares electrophoretic tears patterns of normal subjects and patients with scleroderma with and without clinical manifestations of KCS. Thus, we try to determine diseases| biomarkers that can be used for noninvasive diagnosis of KCS in patients with scleroderma.
Methods: Subjects were: patients with scleroderma without KCS (n=3), patients with scleroderma with KCS (n=2) and a control group comprising healthy volunteers (n=10). Tears were sampled using the Schirmer method and the sample was eluted from the filter paper in 40 microL of elution solution containing sodium dodecyl sulphate, urea, EDTA, beta-mercaptoethanol and bromphenol blue.
Tear proteins were separated by two-dimensional electrophoresis (in the combination of isoelectric focusing with sodium dodecyl sulphate-poliacrylamide gel electrophoresis) and protein bands were stained with silver.
Results: Tear proteins could be separated into more than 20 bands, main components (tear-specific pre-albumin, lactoferrin, lysozyme, secretory immunoglobulin A and immunoglobulin G) being identified using a marker of molecular weight. Isoelectric points of all proteins separated were determined by comparison with isoelectric point standards. The densitometric analysis of electrophoretic lanes was performed with ordinary flat scanner.
The tear protein patterns of patients with scleroderma and KCS are different in number and intensity of spots from those of healthy subjects. Patients with KCS had decreased levels of lysozyme and lactoferrin in tears.

Conclusion: Two-dimensional electrophoretic analysis of tear protein patterns of patients with scleroderma is a fast, reproducible and simple method that provides information for the diagnosis of KCS in these patients.