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. 2016 Sep;17(3):399-406.
doi: 10.1007/s10561-016-9553-x. Epub 2016 Apr 12.

Cytokine expression and ultrastructural alterations in fresh-frozen, freeze-dried and γ-irradiated human amniotic membranes

Adolfo Paolin  1 , Diletta Trojan  2 , Antonio Leonardi  3 , Stefano Mellone  3 , Antonio Volpe  4 , Augusto Orlandi  4 , Elisa Cogliati  2
Affiliations

Cytokine expression and ultrastructural alterations in fresh-frozen, freeze-dried and γ-irradiated human amniotic membranes

Adolfo Paolin et al. Cell Tissue Bank. 2016 Sep.

Abstract

The aim of this work was to compare the effects on human amniotic membrane of freeze-drying and γ-irradiation at doses of 10, 20 and 30 kGy, with freezing. For this purpose, nine cytokines (interleukin 10, platelet-derived growth factor-AA, platelet-derived growth factor-BB, basic fibroblast growth factor, epidermal growth factor, transforming growth factor beta 1, and tissue inhibitors of metalloproteinase-1, -2, and -4) were titrated in 5 different preparations for each of 3 amniotic membranes included in the study. In addition, the extracellular matrix structure of each sample was assessed by transmission electron microscopy. While freeze-drying did not seem to affect the biological structure or cytokine content of the different amniotic membrane samples, γ-irradiation led to a significant decrease in the tissue inhibitors of metalloproteinase-4, basic fibroblast growth factor and epidermal growth factor, and induced structural damage to the epithelium, basement membrane and lamina densa. The higher the irradiation dose the more severe the damage to the amniotic membrane structure. In conclusion, the Authors recommend processing amniotic membrane under sterile conditions to guarantee safety at every step rather than final sterilization with γ-irradiation, thereby avoiding alteration to the biological characteristics of the amniotic membrane.

Keywords: Amniotic membrane; Cytokines; Freeze-drying; Transmission electron microscopy; γ-Irradiation.

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Figures

Fig. 1
Fig. 1
Cytokine concentrations. Cytokine concentrations in different preparations of HAM samples: fresh-frozen, freeze-dried, and sterilized with 10–20–30 kGy γ-irradiation. * indicates p < 0.05, ** indicates p < 0.01
Fig. 2
Fig. 2
Ultrastructure evaluation of differently prepared HAM samples: a, b, c fresh-frozen HAM, d, e, f freeze-dried HAM, g, h, i freeze-dried HAM, 10 kGy irradiated, l, m, n freeze-dried HAM, 20 kGy irradiated, o, p, q freeze-dried HAM, 30 kGy irradiated
See this image and copyright information in PMC

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