A young child formula with Limosilactobacillus reuteri and GOS modulates gut microbiome and enhances bone and muscle development - a randomized trial

Sep 30, 2026

Uploaded by Riley

A randomized, double-blind controlled trial evaluated whether an experimental young child formula (EYCF) containing Limosilactobacillus reuteri DSM 17938 and galactooligosaccharides (GOS) would improve toddler musculoskeletal development. A total of 182 Filipino children aged 2–3 years received EYCF or minimally fortified milk (CM) for 6 months. EYCF increased tibia speed of sound and remodeled stool microbiota, with enrichment of L. reuteri and higher bifidobacteria. Metabolomics linked 45 modulated metabolites to tryptophane and indole pathways, associating microbial function with bone quality and muscle strength, supporting gut–musculoskeletal crosstalk during early life.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages15

Reading Time38 min

AccessFree to download

Trial design and participants

Outcomes assessed

Effects on bone and muscle development

Tibia speed of sound

Muscle strength and bone markers

Microbiome remodeling

Shotgun metagenomics findings

Microbiota composition and abundance changes

Metabolomics and mechanistic links

Tryptophane/indole metabolism

Background on calcium, vitamin D, and the gut-bone axis

Article [https://doi.org/10.1038/s41467-025-66930-2](https://doi.org/10.1038/s41467-025-66930-2) A young child formula with Limosilactobacillus reuteri and GOS modulates gut microbiome and enhances bone and muscle development: a randomized trial Received: 15 April 2025 | Accepted: 17 November 2025 | | --- | | | | Check for updates | A list of authors and their afﬁliations appears at the end of the paper In this randomized, double-blind controlled trial, 182 Filipino children aged 2-3 years received either an experimental young child formula (EYCF) containing a combination of Limosilactobacillus reuteri DSM 17938 and galactooligosaccharides (GOS; n = 91) or a minimally fortiﬁed milk (CM; n = 91) for 6 months. Primary outcome was tibia speed of sound and secondary outcomes were muscle strength, blood vitamin D levels, bone turnover markers, gut microbiota, fecal calcium fatty acid soaps and gastro-intestinal tolerance. Compared to CM, those in the EYCF group showed increased tibia speed of sound after3and6months. The intervention remodeled the stool microbiome composition, assessed by shotgun metagenomics, with enrichment of L. reuteri and higher biﬁdobacteria presence in the EYCF group. Increased L. reuteri abundance after 6 months of EYCF consumption associates with higher bone quality and muscle strength. Stool metabolomics show 45 metabolites modulated by EYCF consumption and associated to microbiome compositional changes, leading to enrichment of tryptophane and indole metabolism. In summary, consumption of EYCF containing a L. reuteri + GOS synbiotic improves musculoskeletal development in toddlers via modulation of microbiota composition and function. These results provide insights on gutmusculoskeletal crosstalk during early life. Clinicaltrial.gov NCT04799028. Bone is a “dynamic” and highly specialized connective tissue, providing mechanical support for muscular activity and physical protection to tissues and internal organs, as well as a repository for the systemic mineral homeostasis1. Bone tissue is constantly renewed in adults as well as in children and adolescents.“Strong bones” are deﬁned by two different aspects of bone geometry (length & width) and by the amount of mineral deposited in the bone tissue. It is interesting to note that osteoblasts ﬁrst synthesize and excrete the bone matrix proteins (osteoids), which later on, at a short distance from the bone-forming cells, mineralize until about 70% of the bone, with the remaining 30% at a longer distance (6–12 months later). Therefore, bone could rapidly be undermineralized if the mineralization rate is reduced compared to the velocity growth2. Most data seem to conﬁrm that an adequate intake of calcium is important in the mineralization process to reach skeletal maturity. [e-mail:](e-mail: marienoelle.horcajada@rdls.nestle.com)[ marienoelle.horcajada@rdls.nestle.com](e-mail: marienoelle.horcajada@rdls.nestle.com) Prospective randomized clinical trials have demonstrated that calcium supplementation may increase bone mass acquisition in children, adolescence and adulthood up to the third decade of life3,4. When calcium supplementation ceases, the beneﬁcial effect on bone accretion seems to disappear5–7. Inadequate intake of calcium may contribute to failure to develop strong bones during early life5. Therefore, daily recommended calcium intakes have been established by various authorities to meet the needs for growth and bone development in children5. In addition, nutrients (such as vitamin D, long-chain saturated fatty acids (LCSFAs)), which help calcium incorporation into bone, may also contribute to bone development5,8–12. Early life and toddlerhood are crucial windows for bone growth, but they are also critical for microbiome development13. The inﬂuence of the microbiome on bone (re)modeling has been demonstrated through different mechanisms of action, such as modulation of mineral absorption, immune cell activity, and production of micr

FAQs

What intervention was tested in the trial?

The study compared an experimental young child formula containing Limosilactobacillus reuteri DSM 17938 plus GOS against a minimally fortified milk for 6 months.

Which outcome measured the primary effect on bone?

The primary outcome was tibia speed of sound, with improvements observed in the EYCF group after 3 and 6 months.

How did the formula affect the gut microbiome?

EYCF remodeled stool microbiome composition, enriching L. reuteri and showing higher bifidobacteria presence, assessed using shotgun metagenomics.

What metabolomic changes were linked to microbiome function?

Stool metabolomics identified 45 metabolites modulated by EYCF, associated with compositional changes and enrichment of tryptophane and indole metabolism.

Sep 30, 2026

Uploaded by Riley