Stomatological Hospital and College, Key Lab. of Oral Diseases Research of Anhui Province, Anhui Medical University, Hefei, Anhui, China.
State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Clinical Research Center for Oral Diseases, Department of Pediatric Dentistry, School of Stomatology, The Fourth Military Medical University, XI''an, Shaanxi, China.
Front Cell Infect Microbiol. 2024 May 8;14:1368684. doi: 10.3389/fcimb.2024.1368684. eCollection 2024.
Quorum-quenching enzyme Est816 hydrolyzes the lactone rings of -acyl homoserine lactones, effectively blocking the biofilm formation and development of Gram-negative bacteria. However, its applications in the oral field is limited. This study aimed to evaluate the efficacy of enzyme Est816 in combination with antibiotics against periodontitis induced by and .
The antimicrobial efficacy of enzyme Est816 in combination with minocycline, metronidazole, and amoxicillin was determined using the minimum inhibitory concentration test. The anti-biofilm effect of enzyme Est816 was assessed using scanning electron microscopy, live/dead bacterial staining, crystal violet staining, and real-time quantitative PCR. Biocompatibility of enzyme Est816 was assessed in human gingival fibroblasts (HGF) by staining. A rat model of periodontitis was established to evaluate the effect of enzyme Est816 combined with minocycline using micro-computed tomography and histological staining.
Compared to minocycline, metronidazole, and amoxicillin treatment alone, simultaneous treatment with enzyme Est816 increased the sensitivity of biofilm bacteria to antibiotics. Enzyme Est816 with minocycline exhibited the highest rate of biofilm clearance and high biocompatibility. Moreover, the combination of enzyme Est816 with antibiotics improved the antibiofilm effects of the antibiotics synergistically, reducing the expression of the virulence factor leukotoxin gene () and fimbria-associated gene (). Likewise, the combination of enzyme Est816 with minocycline exhibited a remarkable inhibitory effect on bone resorption and inflammation damage in a rat model of periodontitis.
The combination of enzyme Est816 with antibiotics represents a prospective anti-biofilm strategy with the potential to treat periodontitis.
群体感应淬灭酶 Est816 水解酰基高丝氨酸内酯的内酯环，有效阻止革兰氏阴性菌生物膜的形成和发展。然而，其在口腔领域的应用受到限制。本研究旨在评估酶 Est816 与抗生素联合治疗 和 诱导的牙周炎的疗效。
采用最低抑菌浓度试验测定酶 Est816 与米诺环素、甲硝唑和阿莫西林联合的抗菌效果。采用扫描电子显微镜、活菌/死菌染色、结晶紫染色和实时定量 PCR 评估酶 Est816 的抗生物膜作用。采用染色法评估酶 Est816 对人牙龈成纤维细胞（HGF）的生物相容性。建立大鼠牙周炎模型，通过微计算机断层扫描和组织学染色评估酶 Est816 与米诺环素联合的效果。
与米诺环素、甲硝唑和阿莫西林单独治疗相比，同时使用酶 Est816 增加了生物膜细菌对抗生素的敏感性。酶 Est816 与米诺环素联合具有最高的生物膜清除率和良好的生物相容性。此外，抗生素与酶 Est816 的联合使用协同增强了抗生素的抗生物膜作用，降低了毒力因子白细胞毒素基因（）和菌毛相关基因（）的表达。同样，酶 Est816 与米诺环素联合在大鼠牙周炎模型中表现出显著的抑制骨吸收和炎症损伤的作用。
酶 Est816 与抗生素联合是一种有前景的抗生物膜策略，有望治疗牙周炎。
