Kokilakanit Paopanga, Dungkhuntod Nonthakorn, Serikul Nitchadakorn, Koontongkaew Sittichai, Utispan Kusumawadee
Faculty of Dentistry, Thammasat University, Klong Luang, Pathum Thani, Thailand.
International College of Dentistry, Walailak University, Dusit, Bangkok, Thailand.
PeerJ. 2025 Feb 17;13:e18942. doi: 10.7717/peerj.18942. eCollection 2025.
Caffeic acid phenethyl ester (CAPE), a natural phenolic compound, has demonstrated antibacterial effects. Dental caries etiology is multifactorial, including a cariogenic biofilm containing multispecies bacteria. However, the antibacterial property of CAPE on multispecies biofilm is unclear. The aim of this study was to assess the effect of CAPE on the formation and cariogenicity in biofilm containing , , and .
(ATCC 25175), (ATCC 35037), and (ATCC 49456T) were employed in this investigation. Each bacterial strain was cultured in the presence of CAPE, followed by susceptibility assessment through optical density measurements at a 600 nm wavelength. Multispecies biofilm formation was achieved by co-culturing , , and at a 1:1:1 ratio on hydroxyapatite-coated 96-well plates. The anti-adherence activity of CAPE on multispecies biofilm was evaluated using a crystal violet staining assay. Cariogenic gene expression level and glucosyltransferase (GTF) function in CAPE-treated mixed bacteria were evaluated using real-time PCR and enzyme activity assay, respectively. The thickness and bacterial viability in CAPE-treated multispecies biofilm were examined using confocal laser scanning microscopy.
CAPE demonstrated a significant antimicrobial effect on , , and ( < 0.05). The inhibition concentration 50% (IC50) of CAPE against , , and ranged from 1.6-6.4 mg/ml. CAPE significantly hindered the multispecies biofilm adherence ( < 0.05). Furthermore, the expression of genes involved in acidogenicity, aciduricity, sucrose-dependent adhesion and quorum sensing mechanism and GTF activity were significantly decreased in CAPE-treated mixed bacteria ( < 0.05). In a multispecies biofilm, CAPE significantly reduced its thickness and viable bacteria population ( < 0.05). In conclusion, CAPE exhibited antimicrobial, anti-adherence and anti-cariogenic effects within a multispecies biofilm. These findings suggest the potential use of CAPE as an adjunctive anti-cariogenic agent in future dental applications.
咖啡酸苯乙酯（CAPE）是一种天然酚类化合物，已显示出抗菌作用。龋齿病因是多因素的，包括含有多种细菌的致龋生物膜。然而，CAPE对多物种生物膜的抗菌特性尚不清楚。本研究的目的是评估CAPE对含有变形链球菌、远缘链球菌和血链球菌的生物膜形成及致龋性的影响。
本研究采用变形链球菌（ATCC 25175）、远缘链球菌（ATCC 35037）和血链球菌（ATCC 49456T）。每种细菌菌株在CAPE存在的情况下进行培养，随后通过在600nm波长下测量光密度进行药敏评估。通过将变形链球菌、远缘链球菌和血链球菌以1:1:1的比例在羟基磷灰石包被的96孔板上共培养来实现多物种生物膜的形成。使用结晶紫染色试验评估CAPE对多物种生物膜的抗黏附活性。分别使用实时PCR和酶活性测定评估CAPE处理的混合细菌中的致龋基因表达水平和葡糖基转移酶（GTF）功能。使用共聚焦激光扫描显微镜检查CAPE处理的多物种生物膜的厚度和细菌活力。
CAPE对变形链球菌、远缘链球菌和血链球菌显示出显著的抗菌作用（P<0.05）。CAPE对变形链球菌、远缘链球菌和血链球菌 的半数抑制浓度（IC50）范围为1.6 - 6.4mg/ml。CAPE显著阻碍了多物种生物膜的黏附（P<０.05）。此外，在CAPE处理的混合细菌中，参与产酸性、耐酸性、蔗糖依赖性黏附及群体感应机制的基因表达和GTF活性显著降低（P<0.05）。在多物种生物膜中，CAPE显著降低了其厚度和活菌数量（P<0.05）。总之，CAPE在多物种生物膜中表现出抗菌、抗黏附和抗龋作用。这些发现表明CAPE在未来牙科应用中作为辅助抗龋剂的潜在用途。
