Superior performance of biofilm versus planktonic Limosilactobacillus reuteri in protection of the intestines and brain in a piglet model of necrotizing enterocolitis

Abstract

Necrotizing enterocolitis (NEC) is the leading cause of gastrointestinal-related death in premature infants. Its etiology is multifactorial, with intestinal dysbiosis playing a major role. Probiotics are a logical preventative therapy for NEC, however their benefits have been inconsistent. We previously developed a novel probiotic delivery system in which planktonic (free-living) Limosilactobacillus reuteri (Lr) is incubated with biocompatible dextranomer microspheres (DM) loaded with maltose (Lr-DM-maltose) to induce biofilm formation. Here we have investigated the effects of Lr-DM-maltose in an enteral feed-only piglet model of NEC. We found a significant decrease in the incidence of Definitive NEC (D-NEC), death associated with D-NEC, and activated microglia in the brains of piglets treated with Lr-DM-maltose compared to non-treated piglets. Microbiome analyses using 16S rRNA sequencing of colonic contents revealed a significantly different microbial community composition between piglets treated with Lr-DM-maltose compared to non-treated piglets, with an increase in Lactobacillaceae and a decrease in Clostridiaceae in Lr-DM-maltose-treated piglets. Furthermore, there was a significant decrease in the incidence of D-NEC between piglets treated with Lr-DM-maltose compared to planktonic Lr. These findings validate our previous results in rodents, and support future clinical trials of Lr in its biofilm state for the prevention of NEC in premature neonates.

A weighted and cumulative point system for accurate scoring of intestinal pathology in a piglet model of necrotizing enterocolitis

“…a spectrum from hyperemia to necrosis). The 6-grade system has been used extensively in preterm piglet studies and was scored as follows: 1 = normal appearance; 2 = hyperemia; 3 = local hemorrhage; 4 = extensive hemorrhage; 5 = local necrosis and/or pneumatosis intestinalis, and 6 = extensive necrosis and/or pneumatosis intestinalis [8,11,12]. For the systematic, descriptive method, the presence of hyperemia, hemorrhage, pneumatosis intestinalis, and necrosis were registered separately.…”

Advanced Bioinspired Multifunctional Platforms Focusing on Gut Microbiota Regulation

“…In a piglet model of necrotizing small intestinal colitis, enteral administration of Lr in the biofilm state exhibited positive effects on the gut, microbiome, and developing brain, with a greater impact on necrotizing enterocolitis (NEC) compared to Lr in the planktonic state. 313 It could be postulated that biofilm delivery, even without live bacteria, may fulfill similar functions as probiotics, as many of their functionalities are mediated by surface properties and structure. Nevertheless, further research is warranted to validate this hypothesis.…”

“…Gail E. Besner conducted an experiment where L. rhamnosus (Lr) was incubated with biocompatible dextran microspheres (DM) loaded with maltose to induce biofilm formation. In a piglet model of necrotizing small intestinal colitis, enteral administration of Lr in the biofilm state exhibited positive effects on the gut, microbiome, and developing brain, with a greater impact on necrotizing enterocolitis (NEC) compared to Lr in the planktonic state . It could be postulated that biofilm delivery, even without live bacteria, may fulfill similar functions as probiotics, as many of their functionalities are mediated by surface properties and structure.…”