Acceleration of Small Intestine Development and Remodeling of the Microbiome Following Hyaluronan 35 kDa Treatment in Neonatal Mice

Abstract

The beneficial effects of human milk suppressing the development of intestinal pathologies such as necrotizing enterocolitis in preterm infants are widely known. Human milk (HM) is rich in a multitude of bioactive factors that play major roles in promoting postnatal maturation, differentiation, and the development of the microbiome. Previous studies showed that HM is rich in hyaluronan (HA) especially in colostrum and early milk. This study aims to determine the role of HA 35 KDa, a HM HA mimic, on intestinal proliferation, differentiation, and the development of the intestinal microbiome. We show that oral HA 35 KDa supplementation for 7 days in mouse pups leads to increased villus length and crypt depth, and increased goblet and Paneth cells, compared to controls. We also show that HA 35 KDa leads to an increased predominance of Clostridiales Ruminococcaceae, Lactobacillales Lactobacillaceae, and Clostridiales Lachnospiraceae. In seeking the mechanisms involved in the changes, bulk RNA seq was performed on samples from the terminal ileum and identified upregulation in several genes essential for cellular growth, proliferation, and survival. Taken together, this study shows that HA 35 KDa supplemented to mouse pups promotes intestinal epithelial cell proliferation, as well as the development of Paneth cells and goblet cell subsets. HA 35 KDa also impacted the intestinal microbiota; the implications of these responses need to be determined.

Early Antibiotic Exposure Alters Intestinal Development and Increases Susceptibility to Necrotizing Enterocolitis: A Mechanistic Study

“…Villus heights and crypt depths were measured using NIH ImageJ software from at least 50 well-oriented villi and crypts in three animals per group. For determination of Paneth and goblet cell numbers, sections were stained with Periodic Acid Schiff (PAS)/Alcian blue (AB) (MilliporeSigma, St. Louis, MO, USA), as previously described [ 28 ].…”

“…DNA quantity and quality were characterized using the NanoDrop Lite Spectrophotometer (ThermoFisher Scientific). PCR amplification of the hypervariable V3–V4 segment of the 16S rRNA gene was achieved using an Illumina kit, as previously described [ 28 ]. Libraries were sequenced on the Illumina MiSeq platform using the MiSeq Reagent Kit v3 for 600 cycles to collect 300 bp paired-end reads.…”

Immunoregulation of bovine lactoferrin together with osteopontin promotes immune system development and maturation

“…The design of infant formula can be taken in the lactation period and also when infants are weaned, to strengthen the immunity and protect against pathogenic conditions. Mice are given LF and OPN from the 7 th day according to predecessors’ research 49 and weaned until the 21 st day to the 28 th day after birth. 50 The 21 st day is chosen as the end point of the early life stage according to previous studies.…”

S-layer protein 2 of vaginal Lactobacillus crispatus 2029 enhances growth, differentiation, VEGF production and barrier functions in intestinal epithelial cell line Caco-2

“…Having the capacity to inactivate LPS in vivo, ALP is vital in preventing the translocation of LPS, the pro-inflammatory stimulus originated from NEC-inducing proteobacteria [79,80]. Several molecules have been described to be involved in the induction of ALP activity, such as hyaluronan from human milk [81], omega-3, whey protein, and the short chain fatty acid-butyrate [78,82,83]. The presence of human milk oligosaccharides (HMOs) in human milk promote the growth of Bifidobacterium genus that have a role in neonatal intestinal maturation and immune tolerance [84] and LPS-suppressing bacteria [78].…”