Department of Oral Health Sciences, BIOMAT & UZ Leuven, Dentistry, KU Leuven, Kapucijnenvoer 7, 3000, Leuven, Belgium.
Division of Restorative Dental Sciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong, SAR, China.
Sci Rep. 2024 Nov 4;14(1):26618. doi: 10.1038/s41598-024-78042-w.
In this multi-parameter study, the effect of diverse factors related to adhesive application on the activation of host-derived gelatinases was investigated by gelatin zymography, in-situ zymography, fluorogenic DQ-gelatin assay and micro-tensile bond-strength (μTBS) testing. Gelatin zymography disclosed the presence of gelatinases in phosphoric acid-etched dentin powder, while two gold-standard adhesives generated no measurable MMP activation. In-situ zymography revealed that the interfacial gelatinolytic activity from specimens treated with the two adhesives appeared similar as that of the EDTA negative control, indicating no detectable gelatinases were activated upon adhesive treatment. In solution, MMP-2/9 activity significantly decreased upon interaction with both adhesives (two-way linear mixed effects model [LMEM]: p < 0.05); gelatinases were almost completely deactivated upon 1-week incubation at 37 °C (general linear model: p < 0.05); light-curing adhesives increased temperature up to 55 °C, which appeared sufficient to dramatically decrease MMP-2/9 activity (two-way ANOVA: p < 0.05). Finally, challenging adhesive-dentin interfaces with highly concentrated MMP-9 (at a much higher concentration than present in saliva) for 1 m did not significantly affect μTBS (two-way LMEM: p > 0.05). Taken together, the two adhesives did not activate but rather inhibited the release and activation of dentinal gelatinases.
在这项多参数研究中，通过明胶酶谱法、原位酶谱法、荧光 DQ-明胶测定法和微拉伸结合强度（μTBS）测试，研究了与胶粘剂应用相关的多种因素对宿主来源明胶酶激活的影响。明胶酶谱法显示在磷酸蚀刻牙本质粉中存在明胶酶，而两种金标准胶粘剂未产生可测量的 MMP 激活。原位酶谱法显示，用两种胶粘剂处理的标本的界面明胶水解活性与 EDTA 阴性对照相似，表明在胶粘剂处理后未检测到明胶酶的激活。在溶液中，MMP-2/9 活性在与两种胶粘剂相互作用时显著降低（双向线性混合效应模型 [LMEM]：p<0.05）；明胶酶在 37°C 孵育 1 周时几乎完全失活（广义线性模型：p<0.05）；光固化胶粘剂将温度提高到 55°C，这似乎足以显著降低 MMP-2/9 活性（双向 ANOVA：p<0.05）。最后，用高浓度 MMP-9（比唾液中存在的浓度高得多）挑战胶粘剂-牙本质界面 1 分钟，并没有显著影响 μTBS（双向 LMEM：p>0.05）。综上所述，这两种胶粘剂没有激活，而是抑制了牙本质明胶酶的释放和激活。
