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

Adolfo Paolin,Elisa Cogliati,Antonio Leonardi,Stefano Mellone,Augusto Orlandi,Antonio Volpe,Diletta Trojan

doi:10.1007/s10561-016-9553-x

Adolfo Paolin, Elisa Cogliati + Show 5 more

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https://doi.org/10.1007/s10561-016-9553-x

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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.

Highlights

Human amniotic membrane (HAM) has been used in a variety of surgical procedures
Hao et al have shown that human amniotic epithelial and mesenchymal cells both express interleukin-1 receptor antagonist, all the four tissue inhibitors of metalloproteinase (TIMPs), collagen XVIII, and interleukin-10 (Hao 2000)
Our findings show that all the cytokines analyzed were present in fresh-frozen samples and were still present after freeze-drying, whereas sterilization of HAM by exposure to c-radiation led to significant cytokine losses

Summary

Introduction

Human amniotic membrane (HAM) has been used in a variety of surgical procedures. HAM has long been used in ophthalmic surgery, the earliest reported application being in 1940 when De Rotth used fetal membranes to correct symblepharon (De Rotth 1940). Today HAM is widely used for ocular surface reconstruction and treating several important ocular diseases (Paolin et al 2016). All these applications are possible because HAM has anti-inflammatory, antifibrotic properties (Solomon et al 2001; Tseng et al 1999). Hao et al have shown that human amniotic epithelial and mesenchymal cells both express interleukin-1 receptor antagonist, all the four tissue inhibitors of metalloproteinase (TIMPs), collagen XVIII, and interleukin-10 (Hao 2000). Reverse transcriptase–polymerase chain reaction (RTPCR) has shown that HAM expresses several additional cytokines, such as transforming growth factor (TGF-a, -b1, -b2), epidermal growth factor (EGF), keratinocyte growth factor (KGF), basic fibroblast growth factor (bFGF), keratinocyte growth factor receptor (KGFR), hepatocyte growth factor (HGF) and hepatocyte growth factor receptor (HGFR) (Koizumi et al 2000; Li et al 2005; Gicquel et al 2009)

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