Evaluation of Antioxidant and Antibacterial Effects of Lyophilized Cell-Free Probiotic Supernatants of Three Lactobacillus spp. and Their Cytocompatibility Against Periodontal Ligament Stem Cells

Abstract

Background: Periodontitis is a chronic disease characterized by the inflammation of the periodontium and leads to progressive damage, such as gingival atrophy, alveolar bone loss, and tooth loss. Streptococcus mutans and Aggregatibacter actinomycetemcomitans are bacteria that support the occurrence of periodontitis via the ability to form biofilms or damage the alveolar bone and periodontal ligaments. On the other hand, periodontal ligament stem cells (PDLSCs) are cells with differentiation capability into osteoblasts or osteoblasts. Due to their role in periodontal homeostasis and regeneration, PDLSCs are considered to control periodontitis progression. However, probiotics are helpful microorganisms known to have antimicrobial and immune-regulating effects. Objectives: This study aimed to evaluate the antioxidant activity and antimicrobial effects of lyophilized cell-free supernatants (LCFSs) derived from three probiotic strains of Lactobacillus on S. mutans and A. actinomycetemcomitans. Moreover, the effect of these lyophilized supernatants was investigated on the viability and migration capability of PDLSCs. Methods: The antibacterial effects of LCFSs of three probiotic bacteria were investigated by determining the minimum inhibitory concentration and minimum bactericidal concentration. Then, the effect of LCFSs on the survival and migration of PDLSCs was investigated by the MTT method (at 24 and 72 hours) and scratch test (at 0, 24, and 48 hours), respectively. Finally, the antioxidant effect of LCFSs was assessed by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and ferric reducing/antioxidant power methods. Results: The antibacterial properties of different concentrations of acidic and neutral LCFSs derived from three studied probiotic bacteria on S. mutans and A. actinomycetemcomitans were observed within the range of 12.5 - 50% (v/v) (1/8 - 1/2 dilutions with culture medium). Although there were no significant toxic (~ 100% viability) and wound healing effects on PDLSCs when the cells were exposed to either acidic or neutral studied LCFSs in a concentration of 5% (v/v), they showed significant antioxidant activity (~ 90% DPPH inhibition and 0.5 mM Fe2+/L). Conclusions: The results revealed that 5% (v/v) 48-hour acidic and neutral supernatants of three studied probiotics might play a beneficial role in controlling periodontitis.