Faculty of Dentistry, University of Toronto, 124 Edward St, Toronto, ON M5G 1G6, Canada; Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College St, Toronto, ON M5S 3G9, Canada.
Faculty of Dentistry, University of Toronto, 124 Edward St, Toronto, ON M5G 1G6, Canada.
Acta Biomater. 2019 Apr 1;88:325-331. doi: 10.1016/j.actbio.2019.02.033. Epub 2019 Feb 23.
Cholesterol esterase-like (CE) activity from saliva and esterase from cariogenic bacteria hydrolyze ester linkages of dental methacrylate resins. Collagenolytic, matrix metalloproteinase-like (MMP) activities from dentin and bacteria degrade collagen in demineralized tooth dentin. Human neutrophils in the oral cavity contain factors that are hypothesized to have CE and MMP activities that could contribute to the degradation of methacrylate resins and dentinal collagen. OBJECTIVES: To measure the CE and MMP activities from human neutrophils and their ability to degrade dental methacrylate resin composite and dentinal collagen. Neutrophils' CE and MMP activities were measured using nitrophenyl-esters or fluorimetric MMP substrates, respectively. Neutrophils' degradation of resin composite and dentinal collagen was quantified by measuring release of a universal 2,2-Bis[4-(2-hydroxy-3-methacryloxypropoxy)phenyl]propane (bisGMA)-derived resin composite degradation byproduct, bishydroxy-propoxy-phenyl-propane (bisHPPP), or a collagen degradation by-product, hydroxyproline, respectively using ultra performance liquid chromatography/mass spectrometry. Neutrophils' CE activity increased the release of bisHPPP from bisGMA monomer compared to control after 24 and 48 h (p < 0.05). Neutrophils degraded polymerized resin composite and produced higher amounts of bisHPPP than buffer after 48 h of incubation (p < 0.05). Neutrophils show generic MMP, gelatinase, MMP-2 and MMP-9, and collagenase, MMP-1 and MMP-8 activities that were stable or increased over the first 24 h (p < 0.05). Neutrophils degraded demineralized dentin more than buffer-only groups, indicated by higher amounts of hydroxyproline (p < 0.05). The ability of neutrophils to degrade both dental resin composite and tooth dentin, suggest neutrophil's potential role in root caries, and in recurrent carries by accelerating the degradation of resin-dentin interfaces, and compromising the longevity of the restoration. STATEMENT OF SIGNIFICANCE: Neutrophils are part of the innate immune system and are constantly entering the oral cavity through the gingival sulcus, in direct contact with the tooth, restoration, restoration-tooth margins and pathogenic bacteria. The current study is the first to characterize and quantify degradative activities from neutrophils toward methacrylate resin and demineralized dentin, the two main components of the restoration-tooth interface, suggesting that this interface could be negatively influenced by neutrophils, potentially contributing to increase in caries formation and progression, and premature restoration failure. This study provides a significant finding to the biomaterials and oral health fields by identifying a potential weakness in current restorative procedures and materials used to manage gingival proximal and cervical gingival or sub-gingival carious lesions.
胆固醇酯酶样 (CE) 活性来自唾液和致龋细菌中的酯酶,可水解牙甲基丙烯酸树脂中的酯键。牙本质和细菌中的胶原酶、基质金属蛋白酶样 (MMP) 活性可降解脱矿牙本质中的胶原蛋白。口腔中的人中性粒细胞含有被假设具有 CE 和 MMP 活性的因子,这些因子可能有助于甲基丙烯酸树脂和牙本质胶原蛋白的降解。目的:测量人中性粒细胞中的 CE 和 MMP 活性及其降解牙甲基丙烯酸树脂复合材料和牙本质胶原蛋白的能力。使用硝基苯酚酯或荧光 MMP 底物分别测量中性粒细胞的 CE 和 MMP 活性。通过使用超高效液相色谱/质谱法分别测量通用 2,2-双[4-(2-羟基-3-甲基丙烯酰氧基丙氧基)苯基]丙烷 (BisGMA) 衍生的树脂复合材料降解副产物双羟丙氧基-苯基-丙烷 (BisHPPP) 或胶原蛋白降解副产物羟脯氨酸的释放来定量评估树脂复合材料和牙本质胶原蛋白的降解。与对照组相比,24 和 48 小时后,中性粒细胞的 CE 活性增加了 BisGMA 单体中 BisHPPP 的释放 (p < 0.05)。与缓冲液相比,中性粒细胞在孵育 48 小时后降解聚合树脂复合材料并产生更高量的 BisHPPP(p < 0.05)。中性粒细胞表现出通用的 MMP、明胶酶、MMP-2 和 MMP-9,以及胶原酶、MMP-1 和 MMP-8 活性,这些活性在最初的 24 小时内是稳定的或增加的 (p < 0.05)。与仅缓冲组相比,中性粒细胞降解脱矿牙本质的能力更高,表现为羟脯氨酸的量更高 (p < 0.05)。中性粒细胞降解牙树脂复合材料和牙本质的能力表明,中性粒细胞在根龋和复发性龋中可能具有潜在作用,通过加速树脂-牙本质界面的降解并损害修复体的寿命,从而加速龋的进展。意义陈述:中性粒细胞是先天免疫系统的一部分,通过牙龈沟不断进入口腔,与牙齿、修复体、修复体-牙齿边缘和致病性细菌直接接触。本研究首次对中性粒细胞对甲基丙烯酸树脂和脱矿牙本质的降解活性进行了特征描述和量化,这两种物质是修复体-牙齿界面的主要成分,这表明该界面可能受到中性粒细胞的负面影响,从而可能导致龋病形成和进展的增加,并导致修复体过早失效。本研究为生物材料和口腔健康领域提供了一个重要发现,即确定了当前修复程序和用于管理牙龈近中或颈龈或龈下龋损的材料的潜在弱点。