DeCesaris D S, Hayashi Michael A L, Vickerman M M, Rickard A H, Tenuta L M A
Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, MI, USA.
Department of Oral Biology, School of Dental Medicine, University at Buffalo, Buffalo, NY, USA.
J Oral Microbiol. 2025 Jul 15;17(1):2528561. doi: 10.1080/20002297.2025.2528561. eCollection 2025.
BACKGROUND/OBJECTIVE: Cariogenic biofilms possess a rich extracellular polysaccharide (EPS) matrix, which can reduce the penetration of anticaries agents such as nanoparticle-based technologies. The aim of this study was to assess the potential of dextranase, an EPS-degrading enzyme, to enhance nanoparticle penetration into biofilms.
Commercially available fluorescent nanoparticles (nanospheres, average diameter around 200 nm) were used as a proxy for nanoparticle treatments. Biofilms of fluorescent 3209/pVMCherry were developed over 48 h in 24-well glass bottom microplates, simulating daily feast (tryptic soy broth (TSB) supplemented with 1% sucrose) and famine periods (TSB supplemented with 0.1 mM glucose). Nanoparticles were co-administered to biofilms with either dextranase (10 U/mL) or pH 6.5 phosphate buffer (placebo). Time-lapse confocal laser scanning microscopy was used to capture six image stacks over approximately 60 minutes of nanoparticle movement through the biofilm. In-house-developed quantitative image analysis methods assessed nanoparticle penetration.
Nanoparticle signal intensity and overlapping signal with cells increased in the presence of dextranase, being significantly higher in the last two CLSM scans compared with the initial one ( < 0.05). Biofilm architecture changed under dextranase, increasing the interaction of nanoparticles with biofilm components.
Dextranase showed potential to enhance nanoparticle-based anticaries therapies.
Dextranase increases the penetration of nanoparticles in cariogenic, extracellular polysaccharide-rich dental biofilms.
背景/目的：致龋生物膜具有丰富的细胞外多糖（EPS）基质，这会降低基于纳米颗粒技术等防龋剂的渗透。本研究的目的是评估葡聚糖酶（一种EPS降解酶）增强纳米颗粒渗透进入生物膜的潜力。
使用市售荧光纳米颗粒（纳米球，平均直径约200nm）作为纳米颗粒处理的替代物。在24孔玻璃底微孔板中培养荧光3209/pVMCherry生物膜48小时，模拟日常的饱餐期（补充1%蔗糖的胰蛋白胨大豆肉汤（TSB））和饥饿期（补充0.1mM葡萄糖的TSB）。将纳米颗粒与葡聚糖酶（10U/mL）或pH 6.5的磷酸盐缓冲液（安慰剂）共同施用于生物膜。使用延时共聚焦激光扫描显微镜在纳米颗粒穿过生物膜的大约60分钟内捕获六个图像堆栈。内部开发的定量图像分析方法评估纳米颗粒的渗透情况。
在葡聚糖酶存在的情况下，纳米颗粒信号强度以及与细胞的重叠信号增加，与最初的共聚焦激光扫描显微镜（CLSM）扫描相比，在最后两次CLSM扫描中显著更高（<0.05）。在葡聚糖酶作用下生物膜结构发生变化，增加了纳米颗粒与生物膜成分的相互作用。
葡聚糖酶显示出增强基于纳米颗粒的防龋治疗的潜力。
葡聚糖酶增加纳米颗粒在富含细胞外多糖的致龋牙生物膜中的渗透。
