■牙髓和根尖周病的主要原因是细菌侵入根管,这是由于牙齿硬组织的持续破坏。根管治疗过程中感染的有效管理需要有效的冲洗。本研究旨在探讨两种抗菌肽(AMPs)的作用,buCaTHL4B和Im-4,对根管生物膜的体外研究。
■选择两种生物膜(粪肠球菌和核梭杆菌)并厌氧培养。应用以下处理:10μg/mLbuCaTHL4B,10μg/mLIm-4,5μg/mLbuCaTHL4B,5μg/mLIm-4,1μg/mLbuCaTHL4B,1μg/mLIm-4,1%NaOCl,无菌水。每组治疗3分钟。随后,将这两个菌株与10μg/mL的buCaTHL4B共培养,10μg/mLIm-4,1%NaOCl,和无菌水24、48和72小时。使用荧光染色的共聚焦激光扫描显微镜(CLSM)检查生物膜,并计算死亡细菌的百分比。采用定量实时PCR(qRT-PCR)来评估生物膜形成期间细菌比例的变化。
■与1%NaOCl相比,10μg/mLbuCaTHL4B或Im-4对两种生物膜表现出明显更大的杀菌作用(p<0.05),导致他们选择进行后续实验。在48小时内,10μg/mLIm-4显示出比buCaTHL4B更强的抗生物膜作用(p<0.05)。经过24小时的生物膜形成期,无菌水组核仁F.的比例降低,粪肠球菌的比例增加。在buCaTHL4B和1%NaOCl基团中,核仁F.的比例低于粪肠球菌(p<0.05),而在Im-4组中,核仁F.的比例高于粪肠球菌(p<0.05)。治疗24h后,两个AMPs组的细菌比例逐渐稳定。
■buCaTHL4B和Im-4在体外表现出对病原根管生物膜的显着抗菌和抗生物膜能力,表明它们作为有前途的添加剂的潜力,以优化作为替代灌溉剂的根管治疗的有效性。
UNASSIGNED: The primary cause of pulp and periapical diseases is the invasion of bacteria into the root canal, which results from the continuous destruction of dental hard tissues. Effective management of infections during root canal therapy necessitates effectively irrigation. This study aims to investigate the effects of two antimicrobial peptides (AMPs), buCaTHL4B and Im-4, on root canal biofilms in vitro.
UNASSIGNED: Two-species biofilms (Enterococcus faecalis and Fusobacterium nucleatum) were selected and anaerobically cultivated. The following treatments were applied: 10 μg/mL buCaTHL4B, 10 μg/mL Im-4, 5 μg/mL buCaTHL4B, 5 μg/mL Im-4, 1 μg/mL buCaTHL4B, 1 μg/mL Im-4, 1% NaOCl, and sterile water. Each group was treated for 3 min. Subsequently, the two strains were co-cultured with 10 μg/mL buCaTHL4B, 10 μg/mL Im-4, 1% NaOCl, and sterile water for 24, 48, and 72 h. The biofilms were examined using confocal laser scanning microscopy (CLSM) with fluorescent staining, and the percentages of dead bacteria were calculated. Quantitative real-time PCR (qRT-PCR) was employed to assess the variations in bacterial proportions during biofilm formation.
UNASSIGNED: Compared to 1% NaOCl, 10 μg/mL buCaTHL4B or Im-4 exhibited significantly greater bactericidal effects on the two-species biofilms (p < 0.05), leading to their selection for subsequent experiments. Over a 48-hour period, 10 μg/mL Im-4 demonstrated a stronger antibiofilm effect than buCaTHL4B (p < 0.05). Following a 24-hour biofilm formation period, the proportion of F. nucleatum decreased while the proportion of E. faecalis increased in the sterile water group. In the buCaTHL4B and 1% NaOCl groups, the proportion of F. nucleatum was lower than that of E. faecalis (p < 0.05), whereas in the Im-4 group, the proportion of F. nucleatum was higher than that of E. faecalis (p < 0.05). The proportions of bacteria in the two AMPs groups gradually stabilized after 24 h of treatment.
UNASSIGNED: buCaTHL4B and Im-4 exhibited remarkable antibacterial and anti-biofilm capabilities against pathogenic root canal biofilms in vitro, indicating their potential as promising additives to optimize the effectiveness of root canal treatment as alternative irrigants.