Treatment of necrotizing enterocolitis by conditioned medium derived from human amniotic fluid stem cells.

Joshua S O’Connell,Nassim Farhat,Niloofar Ganji,Ethan Lau,Mashriq Alganabi,Carol Lee,Andrea Zito,Robert Mitchell,Marissa Cadete,Simon Eaton,Ketan Patel,Paolo De Coppi,Steve Ray,Abdalla Ahmed,Bo Li,Agostino Pierro,Benedetta Bussolati

doi:10.1371/journal.pone.0260522

Joshua S O’Connell, Nassim Farhat + Show 15 more

Open Access

https://doi.org/10.1371/journal.pone.0260522

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Abstract

Necrotizing enterocolitis (NEC) is one of the most distressing gastrointestinal emergencies affecting neonates. Amniotic fluid stem cells (AFSC) improve intestinal injury and survival in experimental NEC but are difficult to administer. In this study, we evaluated whether conditioned medium (CM) derived from human AFSC have protective effects. Three groups of C57BL/6 mice were studied: (i) breast-fed mice as control; (ii) experimental NEC mice receiving PBS; and (iii) experimental NEC mice receiving CM. NEC was induced between post-natal days P5 through P9 via: (A) gavage feeding of hyperosmolar formula four-time a day; (B) 10 minutes hypoxia prior to feeds; and (C) lipopolysaccharide administration on P6 and P7. Intra-peritoneal injections of either PBS or CM were given on P6 and P7. All mice were sacrificed on P9 and terminal ileum were harvested for analyses. CM treatment increased survival and reduced intestinal damage, decreased mucosal inflammation (IL-6; TNF-α), neutrophil infiltration (MPO), and apoptosis (CC3), and also restored angiogenesis (VEGF) in the ileum. Additionally, CM treated mice had increased levels of epithelial proliferation (Ki67) and stem cell activity (Olfm4; Lgr5) compared to NEC+PBS mice, showing restored intestinal regeneration and recovery during NEC induction. CM proteomic analysis of CM content identified peptides that regulated immune and stem cell activity. CM derived from human AFSC administered in experimental NEC exhibited various benefits including reduced intestinal injury and inflammation, increased enterocyte proliferation, and restored intestinal stem cell activity. This study provides the scientific basis for the use of CM derived from AFSC in neonates with NEC.

Highlights

Necrotizing enterocolitis (NEC) is one of the most devastating gastrointestinal diseases affecting preterm, low birth weight, and very low birth weight neonates [1]
conditioned medium (CM) derived from human Amniotic fluid stem cells (AFSC) administered in experimental NEC exhibited various benefits including reduced intestinal injury and inflammation, increased enterocyte proliferation, and restored intestinal stem cell activity
This study provides the scientific basis for the use of CM derived from AFSC in neonates with NEC

Summary

Introduction

Necrotizing enterocolitis (NEC) is one of the most devastating gastrointestinal diseases affecting preterm, low birth weight, and very low birth weight neonates [1]. NEC is characterized by intestinal mucosal injury, inflammation, and necrosis often leading to perforation [1]. Modern medicine and critical care have advanced, treatment for NEC remains unchanged. The mortality from NEC has remained unchanged for decades and reported to be as high as 30–50% [2]. Morbidity for NEC survivors remains high including intestinal failure and neurodevelopmental delay [3]. Current medical treatment of suspected or confirmed NEC is supportive unless further deterioration is seen. If this is the case, an operation remains the only option [1]. It is generally agreed that the best treatment practice would involve the development of a medical treatment to avoid an operation and intestinal resection [1]

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