牙科粘合剂对致癌多物种生物膜的抗菌效果

S. Thaweboon, Takashi Saito, Sirilak Mateekusontan, B. Thaweboon
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引用次数: 0

摘要

龋齿或蛀牙是影响全世界人民的最普遍的多因素慢性疾病之一。它被描述为细菌发酵可食用碳水化合物产生的酸性物质对牙齿结构的局部破坏。病变过程始于牙齿表面的细菌生物膜。血链球菌、戈登链球菌和变异链球菌是与口腔健康或疾病相关的生物膜中的主要生物。变异链球菌被认为是釉质脱矿和龋齿形成的主要原因。戈登链球菌(S. gordonii)和血清链球菌(S. sanguinis)属于链球菌群,被认为是与健康相关的口腔共生细菌,对在口腔硬组织上形成生物膜至关重要。牙科粘合剂中已经加入了几种具有多功能特性的材料。最近,一种酸性单体的钙盐(4-甲基丙烯酰氧乙基偏苯三酸钙盐)被加入牙科粘合剂中,并被证明能抑制突变杆状病毒的致龋生物膜形成。然而,使用单一物种模型可能无法模拟口腔生物膜中错综复杂的拮抗和协同关系。因此,本研究评估了对致癌多菌种生物膜的影响。将由 CMET 和 10-甲基丙烯酰氧癸基磷酸二氢钙(MDCP)组成的牙科粘合剂(Bio-Coat CA)涂抹在 96 孔板的平底表面,并用 LED 光固化。然后在 37 °C 下用无菌唾液涂布 45 分钟,以形成微生物附着的获得性胶层。制备含有变异链球菌(Streptococcus mutans ATCC 25715)、血清链球菌(Streptococcus sanguinis ATCC 10556)和戈登链球菌(Streptococcus gordonii ATCC 10558)的多菌种细菌悬浮液,并将其加入唾液涂层孔中。然后,将平板置于 37°C 的 5%二氧化碳环境中培养 24 小时、48 小时和 72 小时,以支持生物膜的形成。使用 WST-8 微生物细胞计数试剂盒(美国 Dojindo 分子技术公司)测定生物膜中的重要细菌数量。所有测试均一式三份,重复三次。统计分析采用 Kruskal-Wallis 和 Dunn 检验。结果表明,与对照组相比,添加了 CMET 的牙科粘合剂在 24 小时、48 小时和 72 小时内均能显著抑制多菌种细菌生物膜的形成。24 小时、48 小时和 72 小时的生物膜抑制率分别为 29.1%、34.7% 和 33.2%。含有 CMET 的牙科粘合剂在生物膜生长 72 小时内显示出良好的多菌种生物膜抑制效果。它是一种很有前途的粘合剂,可用于防止修复部位的二次龋坏。
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Antimicrobial Effect of Dental Adhesive on Cariogenic Multi-Species Biofilm
Dental caries, or tooth decay, is one of the most widespread chronic and multifactorial diseases affecting people worldwide. It is described as the localized destruction of tooth structure by acids produced from bacteria fermentation of edible carbohydrates. The disease process begins within the bacterial biofilm on the surfaces of the teeth. Streptococcus sanguinis, Sreptococcusgordonii, and Streptococcus mutans are the main organisms in the biofilm associated with health or disease conditions in the oral cavity. Streptococcus mutans is believed to be the primary cause of enamel demineralization and the development of dental caries. S. gordonii and S. sanguinis belong to a group of streptococci considered health-related commensal oral bacteria, crucial for forming a biofilm on oral hard tissues. Several types of materials with multipurpose characteristics have been incorporated into dental adhesives. Recently, the calcium salt of an acidic monomer (calcium salt of 4-methacryloxyethyl trimellitic acid, or CMET) has been included in dental adhesive and demonstrated to suppress cariogenic biofilm formation of S. mutans. However, using a single-species model may not mimic the intricate antagonistic and synergistic relations that occur in oral biofilms. Therefore, the effect on cariogenic multi-species biofilm was assessed in this study. Dental adhesive consisting of CMET and 10-methacryloyloxydecyl dihydrogen calcium phosphate (MDCP) (Bio-Coat CA) was spread over the flat-bottom surface of the 96-well plate and LED light-cured. Then it was coated with sterile saliva at 37 °C for 45 min to form an acquired pellicle for microbial attachment. The multi-species bacterial suspension containing Streptococcus mutans ATCC 25715, Streptococcus sanguinis ATCC 10556, and Streptococcus gordonii ATCC 10558 was prepared and added to the saliva-coated well. Then, the plate was incubated at 37°C in a 5% CO2 atmosphere for 24 h, 48 h, and 72 h to support the biofilm formation. The number of vital bacteria in the biofilm was determined with the WST-8 Microbial Cell Counting Kit (Dojindo Molecular Technologies, USA). All tests were done in triplicate and repeated three times. For statistical analysis, Kruskal-Wallis and Dunn’s tests were employed. The results showed that at 24 h, 48 h, and 72 h, dental adhesive with CMET could inhibit the biofilm formation of multi-species bacteria significantly compared with controls. The percentages of biofilm inhibition were 29.1%, 34.7%, and 33.2% at 24 h, 48 h, and 72 h, respectively. Dental adhesive containing CMET displayed favorable multi-species biofilm-inhibiting effects up to 72 h of biofilm growth. It is a promising adhesive for use to prevent secondary caries at the sites of restorations.
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