Search Results (26)

Three-dimensional (3D) scaffold systems have proven instrumental in advancing our understanding of polymicrobial biofilm dynamics and probiotic interactions within the oral environment. Among oral probiotics, Streptococcus salivarius K12 (Ssk12) has shown considerable promise in modulating microbial homeostasis; however, its long-term therapeutic benefits are contingent upon successful and sustained colonization of the oral mucosa. Despite its clinical relevance, the molecular mechanisms underlying the adhesion, persistence, and integration of Ssk12 into the native oral microbiome/biofilm remain inadequately characterized. In this pilot study, we explored the temporal colonization dynamics of Ssk12 and its impact on the structure and functional profiles of salivary-derived biofilms cultivated on melt-electrowritten poly(ε-caprolactone) (MEW-mPCL) scaffolds, which emulate the native oral niche. Colonization was monitored via fluorescence in situ hybridization (smFISH), confocal microscopy, and RT-qPCR, while shifts in community composition and function were assessed using 16S rRNA sequencing and meta-transcriptomics. A single administration of Ssk12 exhibited transient colonization lasting up to 7 days, with detectable presence diminishing by day 10. This was accompanied by short-term increases in Lactobacillus and Bifidobacterium populations. Functional analyses revealed increased transcriptional signatures linked to oxidative stress resistance and metabolic adaptation. These findings suggest that even short-term probiotic colonization induces significant functional changes, underscoring the need for strategies to enhance probiotic persistence. Full article

Background/Objectives: Sarcopenia is characterized by loss of muscle mass and strength and is associated with aging. Recently, its links with the gut–muscle axis have been reported, suggesting that probiotics could influence muscle health. Methods: In the present study, we investigated the protective roles of two lactic acid bacteria strains, Streptococcus salivarius subsp. thermophilus ST-G30 (ST-G30) and Lacticaseibacillus paracasei LPc-G110 (LPc-G110), on skeletal muscle atrophy induced by dexamethasone (DEX) in C2C12 myotubes. Results: Our results demonstrated that ST-G30 significantly alleviated DEX-induced myotube atrophy by increasing the myotubes’ diameter (25.95 ± 1.28 vs. 15.30 ± 0.30 μm, p < 0.01), improving the fusion index (48.35 ± 1.75 vs. 22.16 ± 2.36%, p < 0.0001), and increasing the protein content (1.78 ± 0.02 vs. 1.56 ± 0.01 mg/mL, p < 0.05) and myotube length (0.61 ± 0.05 vs. 0.33 ± 0.01, p < 0.05), whereas LPc-G110 showed no significant effect on these phenotypes (p > 0.05). Transcriptomic analysis reveals that ST-G30 modulates critical signaling pathways and biological processes related to skeletal muscle health. In the current study, KEGG enrichment analysis and WGCNA enabled identification of the PI3K-Akt signaling pathway as a key regulator of these processes, highlighting its essential role in mitigating DEX-induced muscle atrophy. Furthermore, the overlapping DEGs associated with the PI3K-Akt signaling pathway showed strong correlations with muscle atrophy-related indices. Conclusions: These findings underscore the potential of ST-G30 as a promising anti-muscle atrophy supplement and provide valuable insights for developing strategies to prevent and treat glucocorticoid-induced skeletal muscle atrophy. Full article

The oral microbiome, a complex ecosystem of microbes, is crucial for oral health. Imbalances in this ecosystem can lead to various oral diseases. Probiotics, live beneficial bacteria, offer a potential solution by strengthening oral defences. This study aimed to develop and evaluate a novel toothpaste containing Streptococcus salivarius M18, a probiotic strain. After ensuring compatibility with toothpaste ingredients, a stable formulation with desirable properties was created. The toothpaste demonstrated cleaning efficacy and antimicrobial activity against oral pathogens in vitro. A clinical trial involving healthy adults showed that all doses of the probiotic toothpaste significantly increased S. salivarius M18 levels in saliva, with the effect persisting even after discontinuation. These findings suggest that the toothpaste effectively delivers the probiotic to the oral cavity and promotes colonisation. Further research is needed to optimise the formulation and assess its long-term impact on oral health. Full article

Background: This systematic review aimed to synthesize the existing evidence on the use of Streptococcus salivarius (S. salivarius) probiotics as prophylactic or therapeutic tools for pediatric oral, dental, and respiratory diseases. Methods: A comprehensive search was carried out across multiple databases using the following terms: S. salivarius, probiotic, children, pediatric. Results: The systematic literature search identified 613 publications, which were meticulously screened, and, ultimately, 15 suitable citations were included in this systematic review. Three strains of S. salivarius (M18, K-12, 24SMB) were used, and they all demonstrated positive benefits in pediatric pathology. Conclusions: Administration of S. salivarius has benefits, is effective, and is convenient (cost-effective) in pediatric prophylaxis. Oral administration as a chewable tablet or powder of S. salivarius M18 for 3 months is able to reduce the incidence of black stains, plaque, and tooth decay in children. S. salivarius K-12 treatment decreased the occurrence of pharyngeal, recurrent, and streptococcal disease, and the benefits also extend to a reduction of nonstreptococcal diseases, including tracheitis, viral pharyngitis, rhinitis, flu, laryngitis, acute otitis media, and enteritis. Administration of S. salivarius 24SMB as an intranasal spray was able to reduce the risk of acute otitis media in children prone to this condition. Full article

Lactobacillus species are widely recognized for their probiotic potential, focusing on their mechanisms of health benefits and protection. Here we conducted an in vitro investigation of the probiotic potential with a role in microbiome homeostasis of four strains: Lactiplantibacillus plantarum L6 and F53, Ligilactobacillus salivarius 1, and Lactobacillus helveticus 611. A broad spectrum of antibacterial and antifungal activity was determined. The strain-specific inhibition of Staphylococcus aureus, Streptococcus mutans, Escherichia coli, Pseudomonas aeruginosa, and saprophytic/toxigenic fungi makes them promising as protective cultures. DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic) acid) measurements showed that tested samples had strain-specific capacity for scavenging of radicals. The molecular base for the antioxidant potential of two lyophilized forms of active strains was investigated by electron paramagnetic resonance spectroscopy. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, with fractions of the most active postbiotics obtained by SEC-FPLC (fast protein liquid chromatography) analysis, showed a wide variety of effects on the growth of a K562 myeloid leukemia cell line. The IC₅₀ (half-maximal inhibitory concentration) of L. salivarius 1 was determined to be 46.15 mg/mL. The proven in vitro functionality of the selected lactobacilli make them suitable for development of target probiotics with specific beneficial effects expected in vivo. Further investigations on produced postbiotics and safety have to be completed before they can be considered as scientifically proven probiotic strains. Full article

Synbiotics are mixtures of prebiotics and probiotics that enhance the activity of probiotic bacteria when co-administered to provide greater benefits to the host. Traditionally, the synbiotics that have been discovered enhance gut probiotic strains and are nutritionally complex molecules that survive digestive breakdown until they reach the later stages of the intestinal tract. Here, we screened and identified sugars or sugar substitutes as synbiotics for the oral probiotic strains Streptococcus salivarius BLIS K12 and BLIS M18. Using a modified deferred antagonism assay, we found that 0.5% (w/v) galactose and 2.5% (w/v) raffinose were the best candidates for use as synbiotics with BLIS K12 and M18, as they trigger enhanced antimicrobial activity against a range of bacteria representing species from the mouth, gut, and skin. Using reverse transcriptase quantitative PCR, we found that this enhanced antimicrobial activity was caused by the upregulation of the lantibiotic genes salA, salB, and sal9 in either K12 or M18. This led to the conclusion that either 2.5% (w/v) raffinose or 0.5% (w/v) galactose, respectively, are suitable synbiotics for use in conjunction with BLIS K12 and M18 to enhance probiotic performance. Full article

Artepillin C, drupanin, and plicatin B are prenylated phenylpropanoids that naturally occur in Brazilian green propolis. In this study, these compounds and eleven of their derivatives were synthesized and evaluated for their in vitro antimicrobial activity against a representative panel of oral bacteria in terms of their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values. Plicatin B (2) and its hydrogenated derivative 8 (2′,3′,7,8-tetrahydro-plicatin B) were the most active compounds. Plicatin B (2) displayed strong activity against all the bacteria tested, with an MIC of 31.2 μg/mL against Streptococcus mutans, S. sanguinis, and S. mitis. On the other hand, compound 8 displayed strong activity against S. mutans, S. salivarius, S. sobrinus, Lactobacillus paracasei (MIC = 62.5 μg/mL), and S. mitis (MIC = 31.2 μg/mL), as well as moderate activity against Enterococcus faecalis and S. sanguinis (MIC = 125 μg/mL). Compounds 2 and 8 displayed bactericidal effects (MBC: MIC ≤ 4) against all the tested bacteria. In silico studies showed that the complexes formed by compounds 2 and 8 with the S. mitis, S. sanguinis, and S. mutans targets (3LE0, 4N82, and 3AIC, respectively) had energy score values similar to those of the native S. mitis, S. sanguinis, and S. mutans ligands due to the formation of strong hydrogen bonds. Moreover, all the estimated physicochemical parameters satisfied the drug-likeness criteria without violating the Lipinski, Veber, and Egan rules, so these compounds are not expected to cause problems with oral bioavailability and pharmacokinetics. Compounds 2 and 8 also had suitable ADMET parameters, as the online server pkCSM calculates. These results make compounds 2 and 8 good candidates as antibacterial agents against oral bacteria. Full article

Background and objectives: The development of the oral microbiome begins in the prenatal stage. Breast milk contains antimicrobial proteins, microorganisms, metabolites, enzymes, and immunoglobulins, among others; therefore, differences have been noted in the type of microorganisms that colonize the oral cavity of children who are breastfed compared to those who are formula-fed. Our objective was to establish the relationship between breastfeeding, formula feeding, or mixed feeding (breastfeeding and formula) with the presence of S. mutans in a population of children under 6 months of age. Materials and Methods: The patients were recruited from the Child Care Center of Ciudad Juárez, Chihuahua, and from the pediatric dentistry postgraduate clinics of the Autonomous University of Ciudad Juárez; children exclusively fed maternally, with formula, and/or mixed were included. Those who had been fed within the previous hour were excluded. The sample was taken with a smear of the jugal groove using a sterile micro-brush. For the identification of Streptococcus mutans, a culture of Mitis Salivarius Agar (Millipore) was used. Results: 53.3% corresponded to females and 46.7% to males, 36.7% corresponded to maternal feeding, 23.3% corresponded to formula feeding, and 40% corresponded to mixed feeding. In 90% of the infants, the parents indicated that they did not perform oral hygiene. The CFU count showed that infants who were exclusively breastfed had an average of 9 × 10 CF/mL, formula-fed infants had an average of 78 × 10 CFU/mL, and those who had mixed feeding 21 × 10 CFU/mL. Conclusions: According to the results obtained, it was possible to corroborate that exclusive breastfeeding limits the colonization of Streptococcus mutans compared to those infants who receive formula or mixed feeding; these results could have a clinical impact on the dental health of infants by having a lower presence of one of the main etiological factors involved in dental caries and the type of microbiome established in the oral cavity. Full article

Dental plaque bacteria play an important role in the pathogenicity of periodontitis and peri-implantitis. Therefore, antimicrobial agents are one means of treatment. N-chlorotaurine (NCT) as an endogenous well-tolerated topical antiseptic could be of advantage for this purpose. Accordingly, its microbicidal activity against some dental plaque bacteria was investigated at therapeutic concentrations in vitro. In quantitative killing assays, the activity of NCT against planktonic bacteria and against biofilms grown for 48 h on implantation screws was tested. Electron microscopy was used to demonstrate the formation of biofilm and its morphological changes. The killing of planktonic bacteria of all tested species, namely Streptococcus sanguinis, Streptococcus salivarius, Streptococcus oralis, Streptococcus cristatus, Rothia aeria, and Capnocytophaga ochracea, was shown within 10–20 min by 1% NCT in 0.01 M phosphate-buffered saline at 37 °C. Bacteria grown on screws for 24 h were inactivated by 1% NCT after 15–20 min as well, but the formation of biofilm on the screws was visible in electron microscopy not before 48 h. The killing of biofilms by 1% NCT was demonstrated after 30 min (streptococci) and 40 min (R. aeria). As expected, NCT has broad activity against dental plaque bacteria as well and should be further investigated on its clinical efficacy in periodontitis and peri-implantitis. Full article

S. salivarius M18 administration has been proven to provide positive effects on periodontal health; however, there is still no consensus on the optimum duration of probiotic administration. This study aimed to evaluate the effect of three months of probiotic supplementation on bleeding on probing, signs of gingival inflammation, and dental biofilm. Sixty-two eligible individuals with gingivitis were enrolled in this placebo-controlled, double-blind trial and randomly allocated to the M18 or control groups. Primary outcomes were changes in gingival condition (gingival index, GI; gingival bleeding index, GBI) after 1, 2, and 3 months of lozenges administration and after a one-month washout. Secondary outcomes included changes in the Quigley–Hein plaque index (modified by Turesky et al.) after 1, 2, and 3 months of lozenges administration and after a washout. In total, 60 individuals completed the study (31 and 29 in the M18 group and the control group, respectively). No severe adverse events were reported. Probiotic supplementation resulted in a significant decrease in gingival bleeding at 1 month (effect size 1.09 [CI95%: 0.55–1.63]), 2 months (effect size 0.78 [CI95%: 0.26–1.30]), and 3 months (effect size 0.67 [CI95%: 0.15–1.18]) and a significant reduction in dental plaque accumulation at 2 months (effect size 0.63 [CI95%: 0.12–1.14]) and 3 months (effect size 0.55 [CI95%: 0.03–1.05]). A three-month supplementation with the probiotic resulted in a significant reduction in gingival bleeding and biofilm accumulation; however, a long-lasting effect is not expected, indicating the need for probiotic intake on a long-term basis. Full article

We aimed to assess the effect of oral probiotic containing the Streptococcus salivarius M18 strain on gingival inflammation, bleeding on probing, and oral biofilm. Sixty-one consenting participants aged between 18 and 25 with gingivitis were recruited in this double-blind, parallel-group study and randomly divided into the probiotic group (n = 31) and the placebo group (n = 30). Fifty-seven participants completed the entire study protocol, 27 in the probiotic group and 30 in the placebo group. The outcomes were assessed after 4 weeks of intervention and 4 weeks of follow-up. There was a significant decrease in the Gingival Index, with the effect size of 0.58 [95%CI 0.05–1.10], and Turesky modification of the Quigley and Hein Plaque Index, with the effect size of 0.55 [95%CI: 0.02–1.07], in the probiotic group after the intervention. However, after a 4-week follow-up, the only significant treatment outcome was improved gingival condition according to the Gingival Index. The Gingival Bleeding Index also decreased significantly in the probiotic group after the intervention period; after the follow-up, this parameter did not differ significantly in both groups from the baseline values. In the placebo group, there were no significant improvements in the assessed parameters throughout this study. No serious side effects were registered. Within the limitations of this study, we conclude that the use of oral probiotic containing the Streptococcus salivarius M18 strain resulted in a significant improvement in gingival condition and oral hygiene level in young adults with gingivitis. Trial registration NCT05727436. Funding: none. Full article

Some species of oral bacteria can reduce dietary nitrate to nitrite, which can later be converted to nitric oxide in the nitrate—nitrite—nitic oxide pathway. Increasing nitric oxide availability can reduce blood pressure (BP) and improve exercise performance. Streptococcus salivarius M18 (Streptococcus salivarius M18) is a bacteriocin-producing probiotic that is known to improve oral health by inhibiting pathogenic oral bacteria. However, it is presently unclear whether probiotic-induced alterations to the oral microbiome will influence circulating levels of nitric oxide metabolites and BP. Purpose: To determine the effects of Streptococcus salivarius M18 supplementation on plasma and salivary nitrate and nitrite levels and BP. Methods: Ten healthy males (age 32 ± 8 y, body mass 88.2 ± 15.1 kg) completed 2 × 14-day supplementation phases in a randomized order at least 14 days apart. In one phase, participants consumed Streptococcus salivarius M18 probiotic lozenges (2.5 billion colony-forming units/dose) once per day, and in the other, they ingested water (placebo). The abundance of bacteria on the tongue was assessed via Illumina 16S rRNA gene sequencing, unstimulated saliva, and venous blood samples were collected, and BP was measured pre and post each phase. Saliva and plasma were analysed for nitrate and nitrite using chemiluminescence, and pH was measured in saliva. The change in each outcome from pre- to post-supplementation was compared between phases using repeated measures ANOVA. Results: Plasma nitrite increased from baseline following probiotic supplementation (from 173 ± 39 to 223 ± 63 nM, p = 0.003, 95% CI 192–250 nM). In comparison, there was no change in the placebo phase or between baselines (all p > 0.05). The abundance of nitrite-producing bacteria was not altered, salivary nitric oxide metabolites and pH did not change, and the increase in plasma nitrite did not result in reductions in BP (all p > 0.05). Conclusions: Supplementation with Streptococcus salivarius M18 increased plasma nitrite, a key marker of NO availability. Despite this, Streptococcus salivarius M18 did not lower BP in these healthy normotensive participants. Additionally, the increase in plasma nitrite was not associated with abundance changes in bacteria thought important to NO generation. Further research is required to determine the mechanism behind the increase in plasma nitrite and the potential therapeutic and ergogenic benefits of Streptococcus salivarius M18 supplementation. Full article

Recently, the demand for goat and sheep cheese has increased mainly because of its nutritional and health benefits. As a result, an enormous amount of whey from various animal species is produced as a waste/by-product. The production of functional probiotic fermented beverages from different types of whey protein concentrates (WPC) could be a good way to valorize whey. Meanwhile, reduced environmental pollution and economic sustainability will be provided. In this study, probiotic beverages enriched with 1% kiwi powder were produced from goat, sheep, and cow WPC (15%). Moreover, Streptococcus salivarius subsp. thermophilus and the probiotic bacteria Lactobacillus acidophilus and Bifidobacterium animalis subsp. lactis were used for fermentation. The results showed that WPC significantly increased the protein content and acidity of beverages (p < 0.05). Production with WPC also improved the viability of probiotic bacteria and S. thermophilus, total phenolic compound (TPC), and antioxidant activity of beverages. The highest viability of probiotic bacteria (9.67 log CFU/mL for Bb-12 and, 9.35 log CFU/mL for L. acidophilus) was found in beverages produced from goat WPC. In addition, WPC increased the free amino acid content of beverages, and the highest essential amino acids and branched-chain amino acids were found in beverages produced from goat WPC as 146.19 mg/100 g and 70.31 mg/100 g, respectively (p < 0.05). Consequently, while production with goat, cow, and sheep WPC improved quality compared to the control, beverages produced from goat WPC excelled. The production of a functional probiotic beverage with goat WPC is promising for dairy technology. Full article

The current trend toward using natural food additives, cosmetics, and medicines has motivated industries to substitute synthetic compounds for natural products. Essential oils (EOs) from medicinal plants are a well-known source of chemical compounds that display several interesting biological activities, including antimicrobial action. In this study, we investigated the antibacterial activity of EOs extracted from three Piperaceae species collected in the Brazilian Amazon region against a representative panel of cariogenic bacteria. The minimum inhibitory concentration (MIC) of the essential oils extracted from Peperomia pellucida (PP-EO), Piper marginatum (PM-EO), and Piper callosum (PC-EO) was determined against Streptococcus mutans, S. mitis, S. sanguinis, S. salivarius, S. sobrinus, Enterococcus faecalis, and Lactobacillus casei by using the microplate microdilution method. PM-EO, PC-EO, and PP-EO displayed antibacterial activity against all the tested cariogenic bacteria. PM-EO displayed the best inhibitory activity, with MIC values ranging from 50 to 500 µg/mL. The lowest MIC values were obtained for PM-EO against S. mitis (MIC = 75 μg/mL), Lactobacillus casei (MIC = 50 μg/mL), and S. mutans (MIC = 50 μg/mL). Gas chromatography mass spectrometry (GC-MS) analysis allowed the chemical composition of all the EOs to be identified. The main constituents of PM-EO, PC-EO, and PP-EO were 3,4-(methylenedioxy)propiophenone, α-pinene, and dillapiole, respectively. Finally, the compounds that were exclusively detected in PM-EO are highlighted. Our results suggest that PM-EO may be used in products for treating dental caries and periodontal diseases. Full article

This was an in vitro study that aimed to evaluate the antimicrobial effect of the propolis extract caffeic acid phenethyl ester (CAPE) on four different oral microorganisms. Seven different concentrations of CAPE (0.2, 0.5, 1, 1.5, 2, 3, and 4 mg/mL) for use against Staphylococcus aureus, Streptococcus mutans, Streptococcus oralis, and Streptococcus salivarius were determined using minimum inhibition concentration (MIC), minimum bactericidal concentration (MBC), broth microdilution, and well diffusion tests over 1, 3, 6, 12, and 24 h, while NaF at 0.05 percent was used as a positive control. Staphylococcus aureus was most affected by CAPE’s inhibitory effect on bacterial growth, whereas S. mutans was the least affected. S. mutans and S. oralis had similar CAPE MIC and MBC values of 1 mg/mL and 1.5 mg/mL, respectively. The most resistant bacteria to CAPE were S. salivarius and S. aureus, with MIC and MBC values of 3 mg/mL and 4 mg/mL, respectively. S. oralis, followed by S. salivarius, S. mutans, and S. aureus, had the highest viable count following exposure to CAPE’s MBC values, while S. aureus had the lowest. The current results of the inhibitory effect of CAPE on bacterial growth are promising, and the values of both CAPE MBC and MIC against the related four cariogenic bacterial organisms are significant. CAPE can be employed as an adjunct dental hygiene substance for maintaining good oral hygiene, and has a potential therapeutic effect in the field of oral health care. Full article