In vitro Anti-biofilm and Anti-adhesion Effects of Lactic Acid Bacteria- derived Biosurfactants against Streptococcus mutans.

Maryam Pourhajibagher, Abbas Bahador
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Abstract

Introduction: Biosurfactants are naturally occurring compounds with various ap-plications, biodegradable, non-toxic, and effective in different conditions. This study fo-cuses on the extraction and evaluation of biosurfactants produced by five strains of lactic acid bacteria [LAB] for their potential to inhibit biofilm formation and adhesion by Strep-tococcus mutans.

Methods: The strains of LAB-producing biosurfactants such as Lactobacillus salivarius, L. acidophilus, L. plantarum, L. casei, and L. rhamnosus were confirmed by the hemolysis test. The presence of biosurfactants derived from LAB strains and their molecular compo-sition were confirmed, and their cellular toxicity, minimum inhibitory concentration [MIC], and minimum bactericidal concentration [MBC] were investigated. Ultimately, the anti-biofilm and anti-adhesive activities of LAB-derived biosurfactants against S. mutans were determined. Eventually, the effect of biosurfactants on the changes in gene expression associated with biofilm formation of S. mutans was assessed. All the LAB strains used in this study were biosurfactant producers. The LAB-derived bi-osurfactants exhibited no cytotoxicity towards the human gingival fibroblast [HGF] cell line. According to the results, the lowest and highest MIC values were observed in the biosurfactants derived from L. rhamnosus and L. plantarum at 0.78 mg/mL and 6.25 mg/mL, respectively. The MBC values for the biosurfactants derived from L. rhamnosus, L. casei, L. salivarius, L. acidophilus, and L. plantarum were 3.12, 3.12, 6.25, 12.5, and 12.5 mg/mL, respectively. The LAB-derived biosurfactants at MBC concentrations exhib-ited significant inhibitory effects on biofilm formation and adhesion of S. mutans [P<0.05]. The highest anti-biofilm and anti-adhesion activities were attributed to the biosurfactants derived from L. plantarum, which were not significantly different from the 0.2% chlorhex-idine as a positive control group [P>0.05]. Moreover, all biosurfactants could significantly decrease the gene expression level of gtfB [P>0.05].

Results: The study found that LAB-derived biosurfactants exhibit significant anti-adhesion and anti-biofilm activities against S. mutans without any observed cellular toxicity towards HGF cells.

Conclusion: These promising bioactive compounds can be utilized as natural antimicrobial agents and biofilm inhibitors to prevent microbial biofilm formation and adhesion in vari-ous dental applications, offering a safe and effective alternative for controlling dental bio-films and improving oral health outcomes.

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乳酸菌源生物表面活性剂对变形链球菌的体外抗生物膜和抗黏附作用。
生物表面活性剂是天然存在的化合物,具有多种用途,可生物降解,无毒,在不同条件下有效。本研究主要研究了5株乳酸菌[LAB]产生的生物表面活性剂的提取和评价,因为它们具有抑制变形链球菌生物膜形成和粘附的潜力。方法:采用溶血试验对唾液乳杆菌、嗜酸乳杆菌、植物乳杆菌、干酪乳杆菌、鼠李糖乳杆菌等实验室产生物表面活性剂菌株进行鉴定。证实了从LAB菌株中提取的生物表面活性剂的存在及其分子组成,并对其细胞毒性、最小抑菌浓度(MIC)和最小杀菌浓度(MBC)进行了研究。最后,测定了实验室制备的生物表面活性剂对变形链球菌的抗生物膜和抗粘附活性。最后,我们评估了生物表面活性剂对变形链球菌生物膜形成相关基因表达变化的影响。本研究所用的酵母菌均为生物表面活性剂的生产者。实验室衍生的双表面活性剂对人牙龈成纤维细胞(HGF)细胞系没有细胞毒性。结果表明,鼠李糖生物表面活性剂和植物表面活性剂的MIC值最低为0.78 mg/mL,植物表面活性剂的MIC值最高为6.25 mg/mL。鼠李糖乳杆菌、干酪乳杆菌、唾液乳杆菌、嗜酸乳杆菌和植物乳杆菌的生物表面活性剂MBC分别为3.12、3.12、6.25、12.5和12.5 mg/mL。MBC浓度的生物表面活性剂对变形链球菌的生物膜形成和粘附有显著的抑制作用[P0.05]。此外,所有生物表面活性剂均可显著降低gtfB基因表达水平[P>0.05]。结果:研究发现,实验室衍生的生物表面活性剂对变形链球菌具有显著的抗粘附和抗生物膜活性,对HGF细胞没有任何细胞毒性。结论:这些生物活性化合物可作为天然抗菌剂和生物膜抑制剂,在各种口腔应用中防止微生物生物膜的形成和粘附,为控制口腔生物膜,改善口腔健康状况提供了安全有效的选择。
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