Comparative evaluation of the short-chain fatty acids formate, propionate, and valerate on intestinal barrier maturation and bone development in neonatal mice

Abstract

Short-chain fatty acids (SCFAs) are key microbial metabolites that support intestinal and skeletal development, yet their coordinated effects during early life remain poorly defined. In this study, neonatal mice were administered SCFAs for 28 days to evaluate their impacts on growth, intestinal barrier integrity, immune modulation, bone development, and gut microbiota composition. Valerate supplementation significantly increased body weight and intestinal length. It enhanced villus structure, crypt depth, and goblet cell number, alongside upregulation of tight junction and mucin genes, indicating improved barrier function. Valerate and propionate also promoted the expression of interleukin-4 (IL-4) and interleukin-10 (IL-10), and reduced pro-inflammatory cytokines, suggesting an immunomodulatory shift. In the skeletal system, valerate improved microarchitecture, increased bone mineral density (BMD), and upregulated osteogenic genes runt-related transcription factor 2 (Runx2), fibroblast growth factor receptor 1 (FGFR1), and growth hormone receptor (GHR). Microbiota profiling showed enrichment of several genera (e.g., Fructobacillus, Pantoea, and Ralstonia) that correlated with intestinal and bone parameters. Collectively, these data indicate that valerate supplementation is associated with concurrent improvements in neonatal intestinal and skeletal outcomes, accompanied by shifts in microbiota and changes related to the barrier and immune systems; however, causal links among these intermediate steps remain to be established.