Abstract
Purpose:
Cryopreservec amniotic membranes are currently used in ophthalmic surgery. Our purpose is to evaluate the bimolecular profile of 3 solutions prepared with 18 months cryopreserved amniotic membranes by looking for the presence of mRNA of growth factors, cytokines and extracellular matrix constituents.
Methods:
During 18months, we have cryopreserved portions of amniotic membranes at -80°C which were afterwards used in the preparation of 3 different types of solution through 3 distinctive protocols. RT PCR was then performed in order to identify mRNA of Beta-actin, KGF, HGF,IL-1ra, FasLigand, Lumican and α 2-macroglobuline.
Results:
The amniotic membrane extract prepared with trypsination does not seem to contain mRNA of the different tested molecules. On the other hand, amniotic membrane extracts that were exclusively prepared through shredding and sonification contained Lumican mRNA only and seem not to contain growth factor and cytokine mRNAs.
Conclusions:
The results could be explained by loss of the amniotic membrane biomolecular profile through time. This loss seems to be selectively affecting cytokines and growth factors while extracellular matrix products seemed to be less labile. This hypothesis should be verified by another study in order to compare these results with fresh amniotic membrane extracts
Keywords: 543 growth factors/growth factor receptors •
490 cytokines/chemokines •
503 drug toxicity/drug effects