Department of Pediatrics, Division of Neonatology, University of Texas Medical Branch, Galveston, TX, USA.
Department of Obstetrics and Gynecology, Division of Maternal Fetal Medicine, University of Texas Medical Branch, Galveston, TX, USA.
Pediatr Res. 2020 Apr;87(5):847-852. doi: 10.1038/s41390-019-0632-0. Epub 2019 Nov 22.
Fetal swallowing of human amniotic fluid (hAF) containing trophic factors (TFs) promotes gastrointestinal tract (GIT) development. Preterm birth interrupts hAF swallowing, which may increase the risk of necrotizing enterocolitis (NEC). Postnatally, it is difficult to replicate fetal swallowing of hAF due to volume. We aimed to evaluate whether hAF lyophilization is feasible and its effect on hAF-borne TFs.
We collected hAF (n = 16) from uncomplicated pregnancies. hAF was divided into three groups: unprocessed control (C), concentration by microfiltration (F), and by dialysis and lyophilization (L). EGF, HGF, GM-CSF, and TGF-α were measured in each group by multiplex assay. Bioavailability of TFs was measured by proliferation and LPS-induced IL-8 production by intestinal epithelial cells FHs74.
After dialysis/lyophilization, GM-CSF and TGF-α were preserved with partial loss of EGF and HGF. hAF increased cell proliferation and reduced LPS-induced IL-8 production compared to medium alone. Compared to control, dialysis/lyophilization and filtration of hAF increased FHs74 cell proliferation (p < 0.001) and decreased LPS-induced IL-8 production (p < 0.01).
Lyophilization and filtration of hAF is feasible with partial loss of TFs but maintains and even improves bioavailability of TFs measured by proliferation and LPS-induced IL-8 production by FHs74.
胎儿吞咽含有营养因子（TFs）的人羊水（hAF）可促进胃肠道（GIT）发育。早产会中断 hAF 的吞咽，这可能会增加坏死性小肠结肠炎（NEC）的风险。由于容量的原因，出生后很难复制胎儿对 hAF 的吞咽。我们旨在评估 hAF 冻干是否可行及其对 hAF 携带 TFs 的影响。
我们从无并发症的妊娠中收集 hAF（n=16）。hAF 分为三组：未处理的对照组（C）、微孔过滤浓缩组（F）和透析冻干组（L）。通过多重分析测定每组中的 EGF、HGF、GM-CSF 和 TGF-α。通过肠上皮细胞 FHs74 的增殖和 LPS 诱导的 IL-8 产生来测量 TFs 的生物利用度。
透析/冻干后，GM-CSF 和 TGF-α得以保留，但 EGF 和 HGF 部分丢失。与单独的培养基相比，hAF 增加了细胞增殖并减少了 LPS 诱导的 IL-8 产生。与对照组相比，hAF 的透析/冻干和过滤增加了 FHs74 细胞的增殖（p<0.001）并减少了 LPS 诱导的 IL-8 产生（p<0.01）。
hAF 的冻干和过滤是可行的，虽然部分 TFs 丢失，但通过 FHs74 的增殖和 LPS 诱导的 IL-8 产生测量，仍保持甚至提高了 TFs 的生物利用度。
