Abstract A novel strategy for preventing urogenital tract infections is the incorporation of beneficial strains into the design of vaginal probiotic products. This study aimed to incorporate Lacticaseibacillus rhamnosus CRL1332-nanofibres on feminine hygiene devices (tampons and pantyliners) and textile substrate using different electrospinning setups. The use of agitation in the electrospinning collector system proved to be an optimal strategy for coating tampons. Moreover, the setup with an uniaxial translational nozzle and a conventional collector system was appropriate for coating pantyliners and textile substrates. The incorporation of lactobacilli immobilised in nanofibres onto the devices was confirmed by scanning electron microscopy, yielding measurements similar to those of non-immobilised cells (length ∼2,118 nm; width ∼741 nm). The viable cell number of lactobacilli was around 10 7 -10 8 cfu/tampon, pantyliner, or gr textile substrate and was maintained after one-year of storage under refrigeration conditions. L. rhamnosus CRL1332 immobilised nanofibres preserved its acidifying capability. The samples demonstrated chemical and thermal stability over time under refrigerated and vacuum-packed conditions. Different electrospinning systems proved to be novel and suitable for designing functional vaginal products with viable L. rhamnosus CRL1332 through simple and effective steps. These lactobacilli-coated devices could serve as effective vaginal probiotic delivery systems, promoting the reconstitution of the vaginal microbiota and improving feminine hygiene and health.
Postbiotics, the inactivated microorganisms and their products derived from fermentation, represent an evolution in the rapidly growing biotics industry and extend health benefits without the need for bacterial viability. In this study, we explored the potential of a novel oat-based postbiotic to provide health benefits by modulating the host immune system and gut microbiome. Two Lactobacillus species, Lactiplantibacillus plantarum 276 and Lacticaseibacillus rhamnosus GG, were used together to ferment 15% oat flour for 24 h at 37 °C followed by pasteurisation and freeze-drying to produce the oat-based postbiotic (OP). OP (1 mg/ml) was used to pretreat a mammalian intestinal epithelial cell line (Caco-2) for 1 h before stimulation with TNF-α and IL-1β (10 ng/ml each). The immuno-modulatory and barrier-supporting effect of OP were evaluated by measuring transepithelial electrical resistance (TEER), wound healing, and expression of cytokines genes (IL-6 and CXCL-8) as well as in a nematode (Caenorhabditis elegans) model. OP protected intestinal barrier integrity as measured by TEER in Caco-2 cells and in the C. elegans model. IL-6 and CXCL-8 expressions in OP pretreated cells were upregulated and OP promoted significant wound closure within 24 h. OP also markedly modulated the human gut microbiome as demonstrated by a 6.48-fold increase in native Bifidobacterium spp. in the in vitro human faecal model. Findings of this study suggest that OP is a novel, safe, stable, and effective intervention for foundational gut health and overall wellness.
The multi-strain probiotic formulation (MPF - Oxxyslab™) regulates several biological processes, including oxygenation in human and mouse models of neurodegenerative diseases. Its effect on oxidative metabolism of tumour cells has not yet been investigated. Employing a polarographic approach, we examined the effect of a lysate of MPF and its constituent strains on oxidative phosphorylation (OXPHOS) in colorectal cancer cells. Results indicate that MPF lysate selectively inhibits OXPHOS in undifferentiated but not in differentiated Caco-2 cells. Ceramides, produced through bacterial sphingomyelinase activity, mediate the inhibitory effects of MPF lysate, and the efficacy of the lysate of individual MPF strains in blocking cellular respiration varies due to strain-specific differences in ceramide production. Our results demonstrate that MPF lysate exhibits anti-OXPHOS activity in human colorectal carcinoma cells, selectively targeting undifferentiated Caco-2 cells but not differentiated ones. The reliance of colorectal cancer cells on OXPHOS provides a scientific basis for exploring MPF probiotic formulation as a complementary approach to conventional pharmacology.
Newborns with gastroschisis hospitalised in the neonatal intensive care unit (NICU) are at risk for a disrupted gut microbiome. Infants with gastroschisis are particularly vulnerable to a dysbiotic microbiome; they require prolonged parenteral nutrition (PN) due to intestinal dysmotility, which often leads to growth faltering (GF). This pilot study's goals were to (1) compare the gut microbiome in infants with gastroschisis to infants admitted to the NICU without congenital anomalies, (2) identify differences in the gut microbiome between infants with gastroschisis requiring prolonged PN and those who do not, and (3) compare the microbiome in infants with gastroschisis with GF to those without GF. This was a multi-site prospective cohort study including 17 infants born with gastroschisis and 16 infants with a gestational age greater than 34 weeks admitted to the NICU without congenital anomalies (controls). Prolonged PN was defined as more than 28 days. GF was defined as a decline in weight or length z-score from birth to discharge of ≤-0.8. Stool samples were collected weekly during hospitalisation and analysed by shotgun metagenomics to assess bacterial composition, diversity, and function. Gestational age and birth weight were similar in the gastroschisis group and the control group. Infants with gastroschisis showed increased Staphylococcus aureus and decreased Bifidobacterium longum. Those requiring prolonged PN had a reduced abundance of genes in the glucosidase pathway compared to those who did not. Infants with GF showed a lower abundance of genes involved in the NAD-diphosphatase pathway compared to those without GF. Infants with gastroschisis display a distinct microbial composition and function compared to NICU infants without this condition. Among infants with gastroschisis, differences in bacterial functional capacity were observed in those who required prolonged PN and developed GF.
Composition of the gut microbiota, and related short chain fatty acids (SCFAs) production can influence allergic status. Extensively hydrolysed casein (eHC) infant formulas are used for management of cow's milk protein (CMP) allergy; however, effects of eHC on gut microbiota as a potential mechanism contributing to benefits of eHC for infants with allergy are not well understood. This exploratory study aimed to assess the in vitro effects of eHC on infant gut microbial composition and metabolism. Two batches of a specific eHC preparation, along with a mixture of individual amino acids (AA), were first digested in the in vitro TNO Intestinal Model-1 (TIM-1) simulating infant upper gastrointestinal digestion conditions followed by an in vitro colon fermentation model inoculated with faeces from healthy, formula-fed infants. Peptide profile after digestion of the test compounds, as well as effects of the compounds on microbiota community structure and SCFA production were assessed. Overall peptide profiles of the eHC preparations (qualitative, based on liquid chromatography-mass spectrometry) remained largely unaffected by simulated upper gastrointestinal digestion. Faecal fermentation of the specific eHC preparations influenced microbiota composition, with a major impact on Bacteroides abundance. SCFA production was increased beyond control conditions by eHC but not AA. The impact of the AA mixture differed considerably from that of the eHC containing specific peptides. Results suggest that eHC formulas can impact microbiota composition and SCFA production in an in vitro colon fermentation model. While the descriptive results of this study require future confirmation under a more extensive experimental setup, it can be hypothesised that mechanisms modulating microbiota composition and metabolism occurring in infants fed with eHC formula for the management of CMP allergy might contribute to clinical observations related to allergy and tolerance development.
