Aggregation and adhesion ability of various probiotic strains and Candida species: An in vitro study.

Background/purpose: The ability of probiotics to inhibit Candida adhesion is a crucial characteristic that prevents Candida colonization and infection progression. This study aimed to explore aggregation, adhesion, and cell surface characterization of probiotic and Candida strains and to evaluate the effect of probiotics and their cell-free supernatants (CFSs) as postbiotics on Candida adhesion to human oral keratinocytes. Materials and methods: Eight probiotic strains and five reference Candida strains were tested for autoaggregation, coaggregation, adhesin on human oral keratinocytes (H357), and cell surface properties. The anti-Candida adhesion activities of probiotic strains and CFSs were investigated. Results: The results showed that most probiotics exhibited high adhesion to H357 cells, specifically oral probiotic Lacticaseibacillus rhamnosus SD4, Limosilactobacillus fermentum SD7, and L. rhamnosus SD11, and adhesion ability of probiotic strains was strongly related to their autoaggregation, cell surface charges, and hydrophobicity. Candida strains also revealed a high level of adhesion to H357 cells. Candida albicans and C andida glabrata showed significantly higher adhesion abilities than others. After a combination of Candida with probiotics or their CFSs, Candida adhesion was significantly reduced. The anti-Candida adhesion property of probiotics was strongly related to their autoaggregation, coaggregation, and adhesion abilities. Conclusion: This study demonstrated that oral probiotic strains may be useful probiotics for preventing and treating oral candidiasis due to their high ability of aggregation, adhesion, and anti-Candida adhesion to H357 cells.

Effect of mouthwash containing poly l-Lysine and glycerol monolaurate on oral Helicobacter pylori relating to biofilm eradication, anti-adhesion, and pro-inflammatory cytokine suppression.

Background/purpose: Helicobacter pylori has been found to be related to periodontitis, and the oral cavity has been considered a reservoir for H. pylori gastritis infection. Thus, this study evaluated the effect of mouthwash containing poly l-Lysine and glycerol monolaurate on inhibiting H. pylori growth, biofilm formation, cell cytotoxicity, adhesion ability, cagA mRNA expression, and pro-inflammatory cytokines stimulated by H. pylori. Materials and methods: Nineteen H. pylori strains were isolated from the oral cavity. The effectiveness of mouthwash containing poly l-Lysine and glycerol monolaurate was examined for its ability to inhibit H. pylori growth and biofilm formation and was tested for cell viability in oral epithelial cells (H357), gastric adenocarcinoma cells (AGS), and periodontal ligament cells (PDL). Additionally, the mouthwash was tested for reducing cagA mRNA expression, adhesion ability to H357 and AGS cells, and pro-inflammatory cytokines stimulated with H. pylori in AGS and PDL cells. Results: The mouthwash containing poly l-Lysine and glycerol monolaurate could eradicate the biofilm by 14.9-19.9% after incubation at 5 min, and cell viability revealed 77.2, 79.8, and 100.0% for AGS, H357, and PDL cells, respectively. Moreover, the mouthwash containing poly l-Lysine and glycerol monolaurate could down-regulate cagA mRNA expression, reduce adhesion of H. pylori by approximately 9.5-47.8% for H357 cells and 24.5-62.9% for AGS cells, and decrease pro-inflammatory cytokines, especially interleukin-8, stimulated with H. pylori. Conclusion: Mouthwash containing poly l-Lysine and glycerol monolaurate could inhibit H. pylori growth and reduce their virulence expression. The mouthwash also revealed low cytotoxicity to oral and gastric cells.

Anticandidal Activity of Various Probiotic Lactobacillus Strains and Their Efficacy Enhanced by Prebiotic Supplementation.

Probiotics and prebiotics have been considered as alternative approaches for promoting health. This study aimed to investigate the anticandidal potential of various probiotic Lactobacillus strains and their cell-free supernatants (CFSs). The study assessed the impact of inulin and some fruits as prebiotics on the growth of selected probiotic strains in relation to their anticandidal activity, production of short-chain fatty acids, total phenolic content, and antioxidant activity. Results revealed variations in anticandidal activity based on the specific strains and forms of probiotics used. Non-adjusted CFSs were the most effective against Candida strains, followed by probiotic cells and adjusted CFSs (pH 7). Lacticaseibacillus rhamnosus SD4, L. rhamnosus SD11 and L. rhamnosus GG displayed the strongest anticandidal activity. Non-adjusted CFSs from L. rhamnosus SD11, L. rhamnosus SD4 and L. paracasei SD1 exhibited notable anticandidal effects. The adjusted CFSs of L. rhamnosus SD11 showed the highest anticandidal activity against all non-albicans Candida (NAC) strains, whereas the others were ineffective. Supplementation of L. rhamnosus SD11 with prebiotics, particularly 2% (w/v) mangosteen, exhibited positive results in promoting probiotic growth, short-chain fatty acids production, total phenolic contents, and antioxidant activity, and the subsequent enhancing anticandidal activity against both C. albicans and NAC strains compared to conditions without prebiotics. In conclusion, both live cells and CFSs of tested strains, particularly L. rhamnosus SD11, exhibited the best anticandidal activity. Prebiotics supplementation, especially mangosteen, enhanced probiotic growth and beneficial metabolites against Candida growth. These finding suggested that probiotics and prebiotic supplementation may be an effective alternative treatment for Candida infections.

Unlocking the genetic potential of Lacticaseibacillus rhamnosus strains: Medical applications of a promising probiotic for human and animal health.

Lacticaseibacillus rhamnosus is a group of probiotic strains that have gained popularity for their potential health benefits such as promoting digestive health, boosting the immune system, improving lactose digestion, preventing and treating antibiotic-associated diarrhea, reducing the severity and duration of certain infections, and preventing the formation of dental plaque. In particular, L. rhamnosus strains SD4 and SD11 have promising human and animal health applications due to their ability to inhibit the growth of harmful pathogens. This study presents an in silico genomic analysis of L. rhamnosus strains SD4 and SD11. We analyzed draft genomes and conducted comparative genome analyses against several other probiotic strains, aiming to gain insights into the genomes of the two strains and to compare them to related strains isolated from other sources. We also aimed to clarify the functional mechanisms and adaptation of these strains to specific environments. Comprehensive insights into the genomes of L. rhamnosus SD4 and SD11 could enhance our understanding of their capacity to colonize, adapt, and exhibit probiotic properties after administration. This study holds significance in advancing our understanding of the potential health benefits associated with these strains and in elucidating the underlying mechanisms responsible for their effectiveness in humans and animals.

Prevalence and virulence factors of Helicobacter pylori isolated from oral cavity of non-disease, gastritis, and gastric cancer patients.

Background/purpose: The oral cavity is considered a reservoir of Helicobacter pylori associated with gastric infection. It aimed to examine the prevalence of H. pylori strains from the oral cavity and gastric tissue of patients with different stage of gastric-diseases. Strains were further characterized for virulence genes, adhesion ability, and inflammation responses. Materials and methods: 11 non-disease, 15 gastritis, and 15 gastric cancer participated in the study. After clinical examination, gastric biopsies, saliva and plaque samples were collected and H. pylori levels were examined by real-time PCR and cultivation. The cagA and vacA genes were investigated from the culture strains. Adhesion ability and pro-inflammatory responses were analyzed in comparison between the presence of virulent genes and disease status. Results: Relatively poor periodontal condition was found among gastric cancer patients. Prevalence of H. pylori-positive was 84.8% and 19.5% by real-time PCR and cultivation, respectively. The cagA and vacA gene-positive strains were 52.6% and 5.3%, respectively, which were found more in gastric cancer patients. The cagA gene-positive strains were found to be higher in gastric cancer patients, and strains had significantly higher adhesion ability and pro-inflammation expressions than the cagA gene-negative strains. Conclusion: Colonization by H. pylori in oral cavity was confirmed, and the cagA gene-positive strains play a crucial role in both adhesion and inflammatory responses. The presence of H. pylori and its virulence gene in oral cavity should be received attention. An eradication of such strains from oral cavity may help to prevent the transmission and recolonization to gastric organs.

Prevalence and virulence factors of haemolytic Enterococcus faecalis isolated from root filled teeth associated with periradicular lesions: A laboratory investigation in Thailand.

AIM: Previous endodontic research has provided limited understanding of the prevalence and roles of haemolytic and non-haemolytic Enterococcus faecalis strains in root filled teeth. This study aimed to determine the prevalence of these strains in root filled teeth with periradicular lesions and investigate their associated virulence factors. METHODOLOGY: A total of 36 root canal samples were collected from 36 subjects. The prevalence of E. faecalis was determined using culture and PCR methods. Antibiotic susceptibility of haemolytic and non-haemolytic E. faecalis strains was assessed using the broth dilution assay. The cytokine stimulation in periodontal ligament (PDL) cells and neutrophil migration were evaluated using real-time PCR and migration assay, respectively. Cell invasion ability of the strains was assessed using a cell culture model. Additionally, the virulence gene expression of the haemolytic and non-haemolytic strains was investigated using real-time PCR. The Mann-Whitney U and Spearman's ρ tests were used to examine the significant difference between the two strains and to analyse the correlation between phenotype and gene expression, respectively. RESULTS: Enterococcus faecalis was detected in 33.3% and 88.9% of samples by culture and real-time PCR, respectively. Haemolytic strains were found in 36.4% of subjects. Non-haemolytic strains exhibited susceptibility to erythromycin and varying susceptibility to tetracycline, while all haemolytic strains were resistant to both antibiotics. Haemolytic strains significantly upregulated the expression of IL-8, OPG and RANKL in PDL cells (p < .05). Notably, the fold increases in these genes were higher: IL-8 (556.1 ± 82.9 vs. 249.6 ± 81.8), OPG (2.2 ± 0.5 vs. 1.3 ± 0.2) and RANKL (1.8 ± 0.3 vs. 1.2 ± 0.1). Furthermore, haemolytic strains had a greater effect on neutrophil migration (68.7 ± 15.2% vs. 46.9 ± 11.4%) and demonstrated a higher level of internalization into oral keratinocyte cells (68.6 ± 0.4% vs. 33.8 ± 0.5%) (p < .05). They also showed enhanced expression of virulence genes associated with haemolysin, surface proteins, collagen-binding and aggregation substances. Gelatinase activity was only detectable in non-haemolytic strains. CONCLUSIONS: This study revealed that haemolytic strains E. faecalis possessed enhanced abilities in host invasion and a higher abundance of virulence factors, suggesting their potential contribution to more severe disease manifestations.

Periodontal diseases in Thai schoolchildren. Clinical and microbiological observations.

The prevalence of periodontitis among Thai schoolchildren is unknown. In a cross-sectional study, the prevalence and severity of periodontal diseases, in a group of Thai schoolchildren, along with the presence and numbers of bacterial species commonly associated with periodontitis were investigated. A consent form was sent out to 192 schoolchildren in one school (Chanachanupathom School) in Chana, Southern Thailand (in the age range of 12-18 years) and 119 attended for a clinical and microbiological examination. Clinical recordings included number of teeth present, DMFT, plaque index, bleeding index, clinical attachment loss (CAL), and probing pocket depth (PPD). Pooled plaque samples were analyzed with culture and qPCR against bacteria associated with periodontitis. The children had low caries experience (DMFT = 3.2 ± 2.3), poor oral hygiene, high bleeding scores, and 67 (56.3%) had at least one interproximal site with CAL ≥ 1 mm. Thirty-seven (31.1%) of the children were diagnosed with periodontitis stage I, and sixteen (13.4%) were classified as periodontitis Stage II. Aggregatibacter actinomycetemcomitans was sparsely found in all but the healthy clinical groups (gingivitis, periodontitis Stage I and II), while the groups showed a high prevalence of Fusobacterium spp., Prevotella intermedia/nigrescens, and Campylobacter species as well as of the periodontitis-associated species Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. Thai schoolchildren have poor oral hygiene with abundant amounts of plaque and high presence of bleeding. Early onset periodontitis is common but mostly in its mild form and is not associated with the presence of A. actinomycetemcomitans.

Short-term use of Lacticaseibacillus rhamnosus SD11 and the oral microbiome: Low caries RCT study.

OBJECTIVES: The objective of the study was to investigate the effect of short-term lozenges containing Lacticaseibacillus rhamnosus SD11 on cariogenic pathogens and on oral microbiota. MATERIALS AND METHODS: This double-blind, randomized, controlled trial included 121 subjects and was randomly divided into the control and probiotic group. All subjects were blindly administered to receive the control- or probiotic L. rhamnosus SD11 lozenges every day for 4 weeks and then followed up for another 4 weeks. RESULTS: After probiotic consumption, the probiotic group had significantly lower levels of Streptococcus mutans and significantly higher levels of total lactobacilli at 4 and 8 weeks compared with the baseline. The 16S rRNA sequencing revealed an increase in bacterial diversity and beneficial bacteria in the Firmicutes phylum, Bacilli class, and a reduction in the mutans streptococci group in the probiotic group. The opposite results were found in the control group. This study did not find any caries increment, nor did the subjects have any side effects after product consumption. CONCLUSION: With the limitation of a short-time study in low caries children, it showed that L. rhamnosus SD11 could increase beneficial bacteria in the Firmicutes phylum and Bacilli class that might support good oral health in children.

Probiotics of Lacticaseibacillus paracasei SD1 and Lacticaseibacillus rhamnosus SD11 attenuate inflammation and ß-cell death in streptozotocin-induced type 1 diabetic mice.

Probiotics provide health benefits in various aspects and are believed to modulate the immune system by balancing gut microbiota homeostasis, termed the "microbiota-immune axis". Recent evidence supports that several Lactobacillus strains possess glucose-lowering and anti-inflammatory effects in an animal model of type 1 diabetes (T1D). Although probiotics of Lacticaseibacillus paracasei SD1 (SD1) and Lacticaseibacillus rhamnosus SD11 (SD11) exert human oral health benefits by reducing harmful bacterial populations, their clinical application regarding hypoglycemic-related traits as well as the underlying mechanisms are still lacking. In this report, we used multiple low doses of streptozotocin (STZ)-induced diabetic BALB/c mice to explore the effects of SD1 and SD11 supplementation on the regulation of markers related to T1D. Experimental mice were randomly assigned into five groups, non-STZ + V, STZ + V, STZ + SD1, STZ + SD11, and STZ + SDM (mixture of SD1 and SD11), and physiological data were measured every week. Blood and pancreas samples were collected at 4- and 8-weeks. Our results indicate that supplementation with SD1, SD11, or SDM for 8 weeks significantly improved body weights, glycemic levels, glucose tolerance, insulin levels, and lipid profiles. Probiotic administration also preserved islet integrity and increased ß-cell mass in STZ-injected mice, as well as prevented infiltration of macrophages, CD4+, and CD8+ T cells into the islets. Significantly, SD1 and SD11 suppressed the levels of IL1-ß, TNF-α and IFN-γ and increased IL-10, which is concomitant with the inhibition of cleaved caspase 3, caspase 9, caspase 8, proapoptotic Bax, NF-κBp65, pSTAT1, and iNOS. Additionally, the survival ability of ß-cells was mediated by upregulated anti-apoptotic Bcl2. We conclude that SD1 and SD11 attenuate STZ-induced diabetic mice by stabilizing glycemic levels and reducing inflammation, thereby protecting ß-cells. Among the probiotic treatment groups, SD11 revealed the best results in almost all parameters, indicating its potential use for alleviating hyperglycemia-associated symptoms.

Characterization of adhesion, anti-adhesion, co-aggregation, and hydrophobicity of Helicobacter pylori and probiotic strains.

Objectives: To characterize the adhesion ability of nine Helicobacter pylori strains and eight probiotics in human oral keratinocyte cells (H357 cells) in comparison to intestinal cells (Caco-2 and HIEC-6 cells). Subsequently, the anti-adhesion and co-aggregation abilities of the selected probiotic strains on H. pylori strains were investigated. Methods: Nine H. pylori strains, including H. pylori ATCC43504 (type strain), and 8 clinical strains, were isolated from oral samples of three patients (one non-disease, one gastritis patient, and one gastric cancer patient). Eight selected probiotic strains were used, as follows: Lacticaseibacillus paracasei SD1, Lacticaseibacillus rhamnosus SD4, L. rhamnosus SD11, Limosilactobacillus fermentum SD7, L. rhamnosus GG, Limosilactobacillus reuteri ATCC-PTA6475, Lacticaseibacillus casei Shirota, and L. paracasei CNCM I-1572. The adhesion and anti-adhesion abilities of H. pylori and the probiotic strains were investigated in H357, Caco-2, and HIEC-6 cells. Co-aggregation at various pHs, hydrophobicity, and surface receptors of the cell lines for H. pylori strains were examined. Results: All probiotic and H. pylori strains adhered to H357 significantly better than Caco-2, and HIEC-6 cells. Three probiotic strains (SD7, SD4, SD11) showed significantly higher adhesion than others. Of the clinical H. pylori strains, isolates from a gastric cancer patient had the highest adhesion ability to all of the cell lines tested. Probiotic strains that exhibited high adhesion ability provided high anti-adhesion and co-aggregation against H. pylori strains. Acidic conditions encouraged the co-aggregation of probiotics to H. pylori strains. Conclusion: This study provides information relating to the adhesion abilities of clinical H. pylori and probiotic strains to the oral mucosa when compared to the intestinal mucosa. Certain probiotic strains may be useful for the successful eradication of H. pylori infection via anti-adhesion and co-aggregation.

Anti-cancer Properties of Potential Probiotics and Their Cell-free Supernatants for the Prevention of Colorectal Cancer: an In Vitro Study.

This study aimed to characterize the anti-cancer properties of potential probiotics (Lacticaseibacillus paracasei SD1, Lacticaseibacillus rhamnosus SD4, Lacticaseibacillus rhamnosus SD11, and Lacticaseibacillus rhamnosus GG) and their cell-free supernatants (CFS) for the prevention of colorectal cancer (CRC), which including anti-bacterial and anti-inflammation activities against pathogens associated with CRC (Fusobacterium nucleatum, Porphyromonas gingivalis, ETEC, and Salmonella enterica). The expression of human ß-defensin (2-4) and IL-10 after being stimulated with probiotics was also examined. In addition, anti-cancer activity of CFS and probiotic growth under intestinal conditions were determined. An in vitro study was conducted in the Caco-2 and HIEC-6 cells. Results showed that probiotic cells and their CFS displayed different antibacterial activity, and L. rhamnosus SD11 showed the strongest inhibition of the growth of pathogens. Additionally, both probiotic cell walls and their CFS suppressed pro-inflammatory cytokines after being stimulated with pathogens in Caco-2 and HIEC-6 cells. L. paracasei SD1 and L. rhamnosus SD11 showed significantly higher suppression levels than others and also both strains can stimulate highly expression of hBD (2-4) and IL-10. The CFS of L. paracasei SD1 and L. rhamnosus SD11 inhibited significantly high growth of Caco-2 cells but not much in HIEC-6 cells. Furthermore, all probiotics adhered to Caco-2 and HIEC-6 cells, and L. rhamnosus SD4 showed the highest adhesion to both cells. They could survive more than 70% in intestinal conditions. In conclusion, results indicate that potential probiotics tested exhibited various anti-cancer properties, which may be good candidates used as biotherapy for the prevention or to delay the progression of CRC.

Evaluation of Potential Probiotic Properties of Lactobacillus and Bacillus Strains Derived from Various Sources for Their Potential Use in Swine Feeding.

Beneficial effects of probiotics are relevant to the various potential properties of individual strains, and they may also relate to the original sources of the probiotic strains. This study aimed to characterize the potential probiotic properties of the strains originating from various sources for probiotics use in swine feeding. A total of 9 potential probiotic strains, seven lactobacilli and 2 bacilli, were examined for antimicrobial production against swine pathogens, adhesion and anti-adhesion of potential probiotic strains to IPEC-J2 cells, aggregation ability, host defense peptide expression, and hemolytic assay. The results highlight that all strains derived from different sources could exhibit probiotic properties, although different abilities were observed. L. rhamnosus SD11 exhibited the highest inhibitory effect against all pathogens compared to other strains. Bacillus licheniformis KMP-9, B. subtilis KMP-N004, and L. fermentum SD7 gave the highest internalization and that related to high abilities of exclusion, competition, and displacement inhibition to pathogens. Such strains also gave a higher co-aggregation to all pathogens compared to other potential probiotic strains. L. rhamnosus GG, L. fermentum SD7, L. rhamnosus SD4, and B. subtilis KMP-N004 had significantly higher pBD-2 mRNA expression than other strains. None of potential probiotic strains showed hemolytic activity. In conclusion, the strains derived from either humans or animals possessed desirable probiotic properties including inhibition against porcine pathogens, adhesion capacity to porcine enterocytes, anti-adhesion pathogens to porcine enterocytes, and modulated innate immunity. Results indicate that these probiotic strains may be good candidates for use in swine feeding to reduce the risk of infection.

Characterization of Short Chain Fatty Acids Produced by Selected Potential Probiotic Lactobacillus Strains.

Short-chain fatty acids (SCFAs), particularly butyrate, have received considerable attention with regard to their anti-cancer efficacy in delaying or preventing colorectal cancer. Several studies have reported that certain probiotic strains could produce SCFAs; however, different strains yielded different amounts of SCFAs. This study explored the ability to produce SCFAs of the following probiotic strains: Lacticaseibacillus paracasei SD1, Lacticaseibacillus rhamnosus SD4, Lacticaseibacillus rhamnosus SD11, and Lacticaseibacillus rhamnosus GG. L. paracasei SD1 and L. rhamnosus SD11 exhibited high butyrate production, particularly when the strains were combined. The functions of the SCFAs were further characterized; the SCFAs exerted a positive anti-cancer effect in the colon via various actions, including inhibiting the growth of the pathogens related to colon cancer, such as Fusobacterium nucleatum and Porphyromonas gingivalis; suppressing the growth of cancer cells; and stimulating the production of the anti-inflammatory cytokine IL-10 and antimicrobial peptides, especially human ß-defensin-2. In addition, the SCFAs suppressed pathogen-stimulated pro-inflammatory cytokines, especially IL-8. The results of this study indicated that selected probiotic strains, particularly L. paracasei SD1 in combination with L. rhamnosus SD11, may serve as good natural sources of bio-butyrate, which may be used as biotherapy for preventing or delaying the progression of colon cancer.

Efficacy of Moraceae with chlorhexidine mouthwash on the microbial flora of critically ill intubated patients: a randomized controlled pilot study.

Critically ill intubated patients are routinely provided with chlorhexidine (CHX) for their mouth washing, but CHX mouthwash induces several complications. In this study, we aimed to evaluate the efficacy and safety of Moraceae with CHX mouthwash in the reduction of oral bacterial count in critically ill patients and to compare it with CHX-alone mouthwash. This double-blind, randomized, controlled trial included critically ill patients receiving mechanical ventilation. The patients were randomly divided into two groups based on the Modified Beck oral assessment score. The primary endpoint was a reduction in oral bacterial counts after mouth washing on day 1 and day 4. Thirty patients were included in this study; 15 patients received Moraceae with CHX mouthwash and 15 patients received CHX-alone mouthwash. The oral bacterial counts in the Moraceae with CHX group did not differ from the CHX group after mouth washing on day 1 and day 4 of admission. The patients in the CHX group experienced more intolerable taste than those in the Moraceae group (60% vs. 13.3%, P = 0.008). Moraceae with CHX mouthwash had the same effectiveness as CHX alone on bacterial flora but exhibited less intolerable side effects than CHX alone.Trial registration: TCTR20190530003; 30/05/2019.

Effects of potential probiotics on the expression of cytokines and human ß-defensins in human gingival epithelial cells and in vivo efficacy in a dog model.

OBJECTIVES: The aims of this study were to examine the effects of probiotic strains on the expression of cytokines and human ß-defensins-2-4 in human gingival epithelial cells and to investigate the in vivo efficacy of probiotics in a dog model. DESIGN: Selected probiotics, Lacticaseibacillus paracasei SD1, L. rhamnosus SD4, L. rhamnosus SD11 and Limosilactobacillus fermentum SD7, were examined for the expression of human ß-defensins-2-4, and cytokine responses after stimulated by various periodontopathogens. Subsequently, an in vivo study was set as a randomized, double-blind, placebo-controlled intervention in a dog model. A total of 20 dogs with mild gingivitis were randomly assigned to either the probiotic or control group. The effects of probiotics on periodontopathogenic- and cytokine levels were analyzed after 4 weeks of intervention. RESULTS: It showed that all probiotics could induce human ß-defensins-2-4, interleukin-1ß, interleukin-6, interleukin-8, and tumor necrosis factor-α expressions in human gingival epithelial cells; however, a significant difference had been found among the strains. When individual probiotic strain was combined to periodontopathogens, a significant reduction of IL-8 expression was found. Results of the in vivo study demonstrated that the bacterial levels, especially Porphyromonas gingivalis, and the interleukin-8 levels were significantly decreased after receiving the probiotic products compared to the controls. CONCLUSION: The results of the in vitro and in vivo studies in dogs are encouraging to support the effectiveness of a mixture of probiotic treatments for improvement of periodontal health by reducing periodontopathogens and interleukin-8 levels. Therefore, such probiotics preparation could possibly have therapeutic potential in human periodontitis.

Thermoresponsive sol-gel containing probiotic's cell free supernatant for dental caries prophylaxis.

BACKGROUND: Lactobacillus rhamnosus SD11 is a probiotic derived from the human oral cavity and has potential being used for dental prophylaxis. The cell free supernatant (CFS) of L. rhamnosus SD11 has good antimicrobial and antioxidant effects. AIM: This study aimed to incorporate CFS of the probiotic into thermoresponsive copolymers to create a sol-gel formulation. METHODS: The sol-gel formulation was developed using Poloxamer 407 as the main polymer, which was mixed with natural polymers such as gellan gum, sodium alginate, and xyloglucan in different proportions. The sol-gel formulations were characterized based on their physicochemical parameters such as appearance, pH, viscosity, flow-ability in low temperature, antioxidant and antibacterial activity. An in vitro release study was performed using Franz's diffusion cell and the stability was determined under freeze-thaw cycle conditions. RESULTS: The combination of 15% w/v of poloxamer 407 with 0.5% w/v of sodium alginate was the best sol-gel formulation to deliver the CFS of the probiotic. CONCLUSION: This study was successful in creating a sol-gel formulation using a thermoresponsive copolymer, that could efficiently deliver CFS of the probiotic L. rhamnosus SD11.

Impact of Potential Probiotic Lactobacillus Strains on Host Growth and Development in a Drosophila melanogaster Model.

Lactobacillus paracasei SD1, Lactobacillus rhamnosus SD11, and Lactobacillus gasseri SD12 were proposed as potential probiotics for oral health. However, the effects of them on host physiology are still unknown. This study aimed to select strains that can promote host growth and development in monocolonized Drosophila model compared with axenic and the commercial Lactobacillus rhamnosus GG-treated flies. The morphogenesis and growth of axenic Drosophila melanogaster were assessed from embryo to adult stage when provided with each probiotic strain-supplemented food. The colonization and persistence of probiotic in fly gut were also evaluated. The results indicated that axenic condition caused the lowest adult weight and emergence rate. All probiotic groups had higher weight than axenic group. Lact. rhamnosus SD11 group presented high adult emergence rate equivalent to Lact. rhamnosus GG group, which is significantly higher than the others. However, Lact. gasseri SD12 group had significantly lower adult weight and emergence rate when compared with Lact. rhamnosus GG group. The gut probiotics levels were rapidly increased within the first day after receiving probiotics. After probiotic cessation, their number in gut decreased and was maintained at low level, except for Lact. gasseri SD12, which completely vanished since day one. In conclusion, Lact. paracasei SD1, Lact. rhamnosus SD11, and Lact. rhamnosus GG can affect morphogenesis and weight of flies when fed since immature stage and have short period gut colonization. The findings of this study could possibly imply comparable health-promoting effects between Lact. paracasei SD1 and Lact. rhamnosus SD11 to commercial strain in Drosophila model.

Significant elevation of salivary human neutrophil peptides 1-3 levels by probiotic milk in preschool children with severe early childhood caries: a randomized controlled trial.

OBJECTIVES: To determine salivary human neutrophil peptides 1-3 (HNP1-3) levels in caries-free preschool children and in those with early childhood caries (ECC) or severe-ECC, in a daily probiotic group, receiving reconstituted milk with the probiotic Lactobacillus paracasei SD1 once daily; a triweekly probiotic group, receiving the probiotic milk 3 days a week; and a placebo group. MATERIALS AND METHODS: Oral examination and unstimulated whole saliva collection were conducted in 354 children at baseline, 6 months after intervention (T6), and after probiotic discontinuation (T12). Of the 354, adequate volume of saliva samples from 268 children were simultaneously analyzed for Streptococcus mutans and total lactobacilli levels using qPCR and for HNP1-3 levels using ELISA. RESULTS: In the severe-ECC status, significant increases in the median HNP1-3 levels at T12 were found in both daily and triweekly probiotic groups (p < 0.001). The median S. mutans levels in the daily group were significantly decreased at T6 and T12 (p < 0.01), whereas the median total lactobacilli levels were significantly increased at T6 (p < 0.001). Significantly inverse correlations between altered HNP1-3 and S. mutans levels and significant decreases in caries progression were found in both probiotic groups (p < 0.05). CONCLUSIONS: In the severe-ECC status, daily or triweekly consumption of L. paracasei SD1 significantly enhanced salivary HNP1-3 levels, but reduced S. mutans levels, possibly resulting in reduction of caries progression. CLINICAL RELEVANCE: Significant enhancement of salivary HNP1-3 levels by probiotic consumption is associated with reduction in S. mutans levels, consistent with diminished caries progression in children with severe-ECC.

Efficacy of Probiotic Milk for Caries Regression in Preschool Children: A Multicenter Randomized Controlled Trial.

This multicenter double-blind randomized controlled trial aimed to determine the efficacy of daily or triweekly consumption of reconstituted milk powder, containing Lactobacillus paracaseiSD1, in preschool children for caries prevention. A 6-month intervention was conducted in 487 children (aged 37.6 ± 9.2 months) with ≤4 decayed teeth from 8 childcare centers. Using stratified block randomization, participants were randomly allocated into 3 groups, comprising: (1) daily probiotic, receiving probiotic milk once daily, (2) triweekly probiotic, randomly receiving probiotic milk 3 days a week and the placebo milk for the remaining 4 days, and (3) placebo, receiving milk without probiotics. Each tooth surface was assessed for caries status using the modified Nyvad criteria at baseline (T0), 6 (T6) and 12 (T12) months after T0. Study outcomes were caries transition, including caries progression and regression during the T0-T6, T6-T12 and T0-T12 periods. Negative binomial regression with a generalized linear model was used to estimate the caries outcomes, which were reported as incidence rate ratios (IRRs) and 95% confidence intervals (CIs). At baseline, caries prevalence and mean decayed surfaces were 81.9% and 7.29 ± 7.60, respectively. Three groups were balanced at baseline with an overall dropout rate of 25.2%. After adjusting for age, sex, and the number of noncavitated and cavitated caries at baseline, a decreased caries risk during T0-T6 was shown in both the daily and triweekly probiotic groups (adjusted IRR 0.83, 95% CI 0.72-0.96 in both groups) and during T0-T12 (adjusted IRR 0.84, 95% CI 0.76-0.94, and adjusted IRR 0.86, 95% CI 0.76-0.96, respectively) compared to the placebo group. In contrast, significantly increased regressive surfaces were demonstrated during T0-T6 by adjusted IRR 1.76 (95% CI 1.25-2.48) and 2.01 (95% CI 1.42-2.85) and during T0-T12 by adjusted IRR 1.98 (95% CI 1.50-2.61) and 1.92 (95% CI 1.45-2.55) for the daily and triweekly groups, respectively. In conclusion, probiotic milk consumption either daily or triweekly can modestly prevent new caries, but considerably reverse carious lesions, suggesting that a daily or triweekly dose interval is sufficient to reverse carious lesions.

Oral Lactobacillus strains reduce cytotoxicity and cytokine release from peripheral blood mononuclear cells exposed to Aggregatibacter actinomycetemcomitans subtypes in vitro.

BACKGROUND: This study evaluated the effect of oral lactobacilli on the cytotoxicity and cytokine release from peripheral blood mononuclear cells (PBMCs) when exposed to Aggregatibacter actinomycetemcomitans subtypes in vitro. The supernatants and cell wall extracts (CWEs) of eight A. actinomycetemcomitans strains, representing different subtypes, and three Lactobacillus strains were used. The PBMCs from six blood donors were exposed to supernatants and CWEs of A. actinomycetemcomitans or Lactobacillus strains alone or combinations and untreated cells as control. The cytotoxicity was determined by trypan blue exclusion method and IL-1ß secretion by ELISA. TNF-α, IL-6, and IL-8 secretions were measured using Bioplex Multiplex Immunoassay. RESULTS: Supernatants or CWEs from all bacterial strains showed cytotoxicity and IL-1ß secretion and the subtypes of A. actinomycetemcomitans showed generally a significantly higher effect on PBMCs than that of the Lactobacillus strains. Two highly toxic A. actinomycetemcomitans strains (JP2 and JP2-like) induced a higher response than all other strains. When combined, Lactobacillus significantly reduced the toxicity and the IL-1ß secretion induced by A. acinomycetemcomitans. The effect varied between the subtypes and the reduction was highest for the JP2 and JP2-like strains. The Lactobacillus paracasei strain SD1 had a higher reducing effect than the other Lactobacillus strains. This strain had a consistent reducing effect on all subtypes of A. actinomycetemcomitans cytotoxicity, and release of IL-1ß, IL-6, IL-8, and TNF-α from PBMCs of the blood donors. A strong and significant variation in cytokine release between the six blood donors was noticed. CONCLUSIONS: Lactobacillus spp. and L. paracasei SD1 in particular, showed a limited but statistically significant reducing interaction with A. actinomycetemcomitans toxicity and release of cytokines in vitro.