Cell and Tissue Banking

, Volume 13, Issue 2, pp 353–361 | Cite as

The effects of different preservation processes on the total protein and growth factor content in a new biological product developed from human amniotic membrane

Article
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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.

Keywords

Growth factor Human amniotic membrane ELISA Freeze-drying Tissue-powdering Irradiation 
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Notes

Acknowledgments

The authors thank Manuela Voltattorni, (MSc, PhD, Interdepartmental Centre of Biotechnological Research (CIRB) of University of Bologna) for assistance during the ELISA procedure and for the precious suggestions.

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.Eye Bank, Department of OphthalmologyS. Maria della Scaletta HospitalImolaItaly

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