The effects of different preservation processes on the total protein and growth factor content in a new biological product developed from human amniotic membrane |
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Authors: | Alessandra Russo Paola Bonci Paolo Bonci |
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Affiliation: | Eye Bank, Department of Ophthalmology, S. Maria della Scaletta Hospital, Via Montericco 4, Imola, Bologna, Italy. banca.occhi@ausl.imola.bo.it |
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Abstract: | The aim of this work is to quantify the total protein and growth factors content in a tissue-suspension obtained from processed human amniotic membrane (hAM). hAM was collected, frozen, freeze dried, powdered and sterilized by γ-irradiation. At each step of the process, samples were characterized for the total protein amounts by a Bradford protein assay and for the growth factor concentrations by ELISA test of the tissue suspensions. Frozen-hAM samples show higher release of total proteins and specific growth factors in the tissue suspension in comparison with freeze-dried hAM. We observed that even if the protein extraction is hindered once the tissue is dried, the powdering process allows a greater release in the tissue suspension of total proteins and growth factors after tissue re-solubilization in comparison with only the freeze-drying process (+91?±?13% for EGF, +16?±?4% for HGF, +11?±?5% for FGF, +16?±?9% for TGF-β1), and a greater release of EGF (85?±?10%) in comparison with only the freezing process, because proteins become much readily solubilized in the solution. According with these results, we describe a protocol to obtain a new sterile biological product from hAM tissue, with well-known effects of thermal, mechanical and physical processes on the total protein and grow factors contents. |
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