Journal of dentistry · 2025
Effect of direct arginine supplementation on growth of Lacticaseibacillus rhamnosus GG.
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Bijle Mohammed Nadeem, Pudipeddi Akhila, Yiu Cynthia
Аннотация
OBJECTIVES: The study aimed to examine the effect of direct arginine (Arg) supplementation on Lacticaseibacillus rhamnosus GG (LRG) growth.
METHODS: The probiotic LRG ATCC 53103 was routinely cultured in MRS broth and adjusted to 107 CFU/ml. l-Arg at 0.25 % and 0.5 % w/v. was directly supplemented to MRS broth containing LRG. Then, LRG with/without supplemented Arg was characterised to determine growth curves for 72 h under a CO2-driven, environment-controlled spectrophotometer. Whereas, end-point McFarland spectrophotometric, metabolic assays (XTT and WST-8), colony forming units (CFU) assessment, live/dead bacterial confocal imaging, and viable DNA quantification were undertaken after 24 h CO2 incubation to determine LRG growth with/without supplemented Arg.
RESULTS: Growth curve analysis revealed similar absorbance profiles with LRG and LRG+0.25 % Arg with no overlaps to LRG+0.5 % Arg; with a similar trend observed as per end-point McFarland spectrophotometric assessment. The % viability estimated by metabolic assays with LRG and LRG+0.25 % Arg was significantly lower than LRG+0.5 % Arg (p < 0.05). As per CFU assessment, CFU/ml for LRG+0.25 % Arg and LRG+0.5 % Arg was significantly higher than LRG control (p < 0.05). LRG live-dead proportions determined using confocal imaging denote no differences between the groups (p > 0.05), with live cell proportions significantly higher than the dead cells (p < 0.05). However, LRG cellular density appeared to increase with increasing concentrations of Arg. Additionally, the viable cells quantified with PMA-qPCR significantly increased with increasing concentrations of Arg (p < 0.05).
CONCLUSION: Direct supplementation of 0.5 % Arg to LRG enhances LRG growth, demonstrating a promising synbiotic potential. CLINICAL SIGNIFICANCE (48 WORDS): Primary caries prevention is initiated at biofilms using biotic strategies that modulate the microbiome matrix to maintain ecological homeostasis. The results of the present study establish a deliverable synbiotic of Arg and probiotic Lacticaseibacillus rhamnosus GG, demonstrating a promising synergism between interventions in the milieu targeted for high caries-risk individuals.
METHODS: The probiotic LRG ATCC 53103 was routinely cultured in MRS broth and adjusted to 107 CFU/ml. l-Arg at 0.25 % and 0.5 % w/v. was directly supplemented to MRS broth containing LRG. Then, LRG with/without supplemented Arg was characterised to determine growth curves for 72 h under a CO2-driven, environment-controlled spectrophotometer. Whereas, end-point McFarland spectrophotometric, metabolic assays (XTT and WST-8), colony forming units (CFU) assessment, live/dead bacterial confocal imaging, and viable DNA quantification were undertaken after 24 h CO2 incubation to determine LRG growth with/without supplemented Arg.
RESULTS: Growth curve analysis revealed similar absorbance profiles with LRG and LRG+0.25 % Arg with no overlaps to LRG+0.5 % Arg; with a similar trend observed as per end-point McFarland spectrophotometric assessment. The % viability estimated by metabolic assays with LRG and LRG+0.25 % Arg was significantly lower than LRG+0.5 % Arg (p < 0.05). As per CFU assessment, CFU/ml for LRG+0.25 % Arg and LRG+0.5 % Arg was significantly higher than LRG control (p < 0.05). LRG live-dead proportions determined using confocal imaging denote no differences between the groups (p > 0.05), with live cell proportions significantly higher than the dead cells (p < 0.05). However, LRG cellular density appeared to increase with increasing concentrations of Arg. Additionally, the viable cells quantified with PMA-qPCR significantly increased with increasing concentrations of Arg (p < 0.05).
CONCLUSION: Direct supplementation of 0.5 % Arg to LRG enhances LRG growth, demonstrating a promising synbiotic potential. CLINICAL SIGNIFICANCE (48 WORDS): Primary caries prevention is initiated at biofilms using biotic strategies that modulate the microbiome matrix to maintain ecological homeostasis. The results of the present study establish a deliverable synbiotic of Arg and probiotic Lacticaseibacillus rhamnosus GG, demonstrating a promising synergism between interventions in the milieu targeted for high caries-risk individuals.