The effects of different preservation processes on the total protein and growth factor content in a new biological product developed from human amniotic membrane
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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DOI：
10.1007/s10561-011-9261-5
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发表时间：
2012-06-01
影响因子：
1.5
通讯作者：
Bonci, Paolo
中科院分区：
文献类型：
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作者：
Russo, Alessandra;Bonci, Paola;Bonci, Paolo
这项工作的目的是定量从加工的人羊膜(HAM)获得的组织悬浮液中的总蛋白质和生长因子的含量。收集火腿，冷冻，冷冻干燥，粉化，c-射线杀菌。在该过程的每一步，样品的总蛋白含量通过Bradford蛋白分析来表征，生长因子的浓度通过组织悬液的ELISA法来表征。与冻干火腿相比，冷冻火腿样品在组织悬浮液中显示出更高的总蛋白和特定生长因子的释放。我们观察到，即使组织干燥后蛋白质提取受到阻碍，与仅冷冻干燥过程(EGF+91+/-13%，HGF+16+/-4%，成纤维细胞生长因子+11+/-5%，转化生长因子-β1+16+/-9%)相比，粉末过程允许组织再溶解后总蛋白和生长因子在组织悬浮液中释放更多(85+/-10%)，与仅冷冻过程相比，EGF释放更多(85+/-10%)，因为蛋白质更容易溶解在溶液中。根据这些结果，我们描述了一种从火腿组织中获得新的无菌生物制品的方案，该方案具有众所周知的热、机械和物理过程对总蛋白和生长因子含量的影响。
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 c-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-beta 1), 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.