Search

The aim of this work was to study the effect of enzymes such as proteinase K, trypsin, collagenase with hyaluronidase, as well as a mixture of all these enzymes, on albumin and collagens incorporated in the vitreous body, using a cyanine dye as a spectral-fluorescent probe. We studied the vitreous body of the eyes of 19/20-week human fetuses, in which, as we showed earlier, the concentration of albumin in the vitreous body is sufficiently high. Proteinase K steeply decreased the albumin content in the vitreous body, whereas trypsin and hyaluronidase with collagenase had no effect on the albumin content. Collagen was not subjected to proteinase K. Enzymatic digestion of collagen occurred under the action of collagenase with hyaluronidase. The content of albumin and collagen sharply decreased in the system after treatment of the vitreous body with mixture of all enzymes. Hence, the results obtained showed that, even being in the mixture, these enzymes have a selective effect on albumin and collagens. The possibility to study the dose-dependent character of enzymatic vitreolysis using a cyanine dye probe has been shown. The spectral-fluorescent probe for albumin and collagens proved to be useful for experimental approaches at screening the enzymatic mixtures possessing the selective action. The study performed is considered as a preclinical trial, and the method presented as promising for the further research in this field. The effect of the enzymes used for therapeutic purposes on the functional conditions of the vitreous body should be studied.

We used one of cyanine dyes as a spectral and fluorescent probe in the study of the composition of the extracellular matrix of the human eye (its vitreous body). Owing to the unique ability of the dye to bind to collagens and human serum albumin, we revealed the simultaneous presence of both types of biomacromolecules in the vitreous body. The formation of the dye complex with human serum albumin leads to appearance of a long-wavelength absorption band (~612 nm) and a steep rise of fluorescence, whereas in the presence of collagens the dye forms J-aggregates with a longer-wavelength absorption band (640-660 nm) and moderate fluorescence. In this work we studied the composition of the human fetus vitreous body and its dynamics from 9 to 31 gestation weeks. On the basis of the data obtained by this method, we may assume that albumin, being a carrier protein, probably provides the vitreous body and surrounding tissues with necessary growth factors, hormones, lipids, vitamins, and some other biomolecules. The data show that the dye is promising not only for study of albumin functions in eye development, but also for characterization of some eye diseases and for analysis of other extracellular media.