J. Ayivor2, B. Sims1, K. Brawner1, C. Martin1 1University Of Alabama at Birmingham,Department Of Pediatric Surgery,Birmingham, Alabama, USA 2Oakwood University,Department Of Biological Sciences,Huntsville, AL, USA
Introduction: Necrotizing Enterocolitis, (NEC) is the leading cause of intestinal morbidity and mortality in premature infants, characterized by epithelial cell injury and sepsis. Breast milk has been shown to decrease the occurrence of NEC; the exact mechanisms that facilitate this protective process are not clear. Human Milk Oligosaccharides, (HMOs) are complex non-digestible pre-biotic sugars that have been shown to stimulate protective immune responses and beneficial growth of intestinal flora. Exosomes are cell derived proteins found in breast milk known to regulate intracellular signaling, inflammation, and immune responses. The objective of this study was to assess the impact of exosomes and human milk oligosaccharides compared to milk ultrafiltrate on the protection of intestinal epithelial cells. Rat intestinal epithelial cells (IEC-6) were used for experimentation.
Methods: Upon reaching confluence, the IEC-6 were pre-treated for 2 hours with 0.1 µg/ml, 1µg/ml or 10 µg/ml of exosomes, 1µg/ml, 10 µg/ml HMOs, or a 5 µg/ml dose of breast milk ultra-filtrate. Following pretreatment, cells were injured with H2O2 for 2 hours. Cell viability was assessed through trypan blue staining.
Results: An Ordinary ANOVA was used to determine significance. Exosomes and HMOs were found to be protective against cell injury with the p value <0.001.
Conclusion: Ultrafiltrate was not protective against cell injury. It was determined that breast milk is immunologically active and protective against epithelial cell injury. Exosomes and HMOs improved cell viability; breast milk ultrafiltrate did not. This suggests that isolated and concentrated breast milk components may have an added therapeutic benefit. Future studies will further clarify these mechanisms of protection.