Lyophilization of human amniotic fluid is feasible without affecting biological activity

John I. Coon, Sangeeta Jain, Krishna M. Sepuru, Yerin Chung, Krishnan Mohankumar, Krishna Rajarathnam, Sunil K. Jain

Research output: Contribution to journalArticle

Abstract

Background: 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. Methods: 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. Results: 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). Conclusions: 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.

Original languageEnglish (US)
JournalPediatric Research
DOIs
StateAccepted/In press - Jan 1 2019

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Freeze Drying
Amniotic Fluid
Interleukin-8
Deglutition
Dialysis
Granulocyte-Macrophage Colony-Stimulating Factor
Epidermal Growth Factor
Biological Availability
Cell Proliferation
Necrotizing Enterocolitis
Premature Birth
Gastrointestinal Tract
Epithelial Cells
Pregnancy
Control Groups

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

Coon, J. I., Jain, S., Sepuru, K. M., Chung, Y., Mohankumar, K., Rajarathnam, K., & Jain, S. K. (Accepted/In press). Lyophilization of human amniotic fluid is feasible without affecting biological activity. Pediatric Research. https://doi.org/10.1038/s41390-019-0632-0

Lyophilization of human amniotic fluid is feasible without affecting biological activity. / Coon, John I.; Jain, Sangeeta; Sepuru, Krishna M.; Chung, Yerin; Mohankumar, Krishnan; Rajarathnam, Krishna; Jain, Sunil K.

In: Pediatric Research, 01.01.2019.

Research output: Contribution to journalArticle

Coon, John I. ; Jain, Sangeeta ; Sepuru, Krishna M. ; Chung, Yerin ; Mohankumar, Krishnan ; Rajarathnam, Krishna ; Jain, Sunil K. / Lyophilization of human amniotic fluid is feasible without affecting biological activity. In: Pediatric Research. 2019.
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AU - Coon, John I.

AU - Jain, Sangeeta

AU - Sepuru, Krishna M.

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AU - Mohankumar, Krishnan

AU - Rajarathnam, Krishna

AU - Jain, Sunil K.

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N2 - Background: 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. Methods: 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. Results: 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). Conclusions: 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.

AB - Background: 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. Methods: 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. Results: 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). Conclusions: 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.

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