Department of Preventive, Restorative and Pediatric Dentistry, School of Dental Medicine, University of Bern, Freiburgstrasse 7, CH-3010 Bern, Switzerland; Department of Conservative Dentistry & Periodontology, School of Dentistry, Medical University of Vienna, Sensengasse 2a, A-1090 Vienna, Austria.
Department of Preventive, Restorative and Pediatric Dentistry, School of Dental Medicine, University of Bern, Freiburgstrasse 7, CH-3010 Bern, Switzerland.
J Dent. 2018 Nov;78:51-58. doi: 10.1016/j.jdent.2018.08.002. Epub 2018 Aug 3.
Matrix metalloproteinases (MMPs) in dentin and saliva can degrade collagen. Divalent metals are known inhibitors of MMPs, but stannous - such as in the form of stannous chloride (SnCl) or stannous fluoride (SnF) - is yet to be tested for a possible inhibitory effect. In this study, we tested the inhibitory effect on the proteolytic activity of MMP-2 and MMP-9.
Sodium chloride (NaCl), sodium fluoride (NaF), and chlorhexidine (CHX) were used as controls. Gelatin zymography was performed with recombinant human MMP-2 and MMP-9. SnCl, SnF, NaF, NaCl, and CHX were included either in the incubation buffer (M1) or added to the recombinant MMPs (M2) before the MMPs were analyzed using zymography. Furthermore, the effect of SnCl, SnF, and NaF on the enzymatic activity of MMP-2 and MMP-9 was measured in human dentin either before or after acid etching using 37%phosphoric acid. The effect of SnCl, NaF, and CHX on the viability and of SnCl and NaF on the proliferation of human gingival fibroblasts and L929 mouse fibroblasts was also determined.
For M1, inhibitory concentrations (w/v%) of SnCl 0.5% and 0.5%, SnF 0.25% and 0.12%, NaF 0.12% and 0.5%, CHX 0.012% and 0.05%, were observed for MMP-2 and MMP-9, respectively. NaCl had no inhibitory effect. For M2, SnCl 0.007% and 0.12%, and SnF 0.03% and 0.5%, inhibited MMP-2 and MMP-9, respectively. NaF, NaCl and CHX had no effect. The enzymatic activity was slightly reduced when SnCl and NaF were applied on dentin before the acid attack. Regarding cell viability and proliferation of the cells after stimulation with the respective substances, NaF showed almost no effect, SnCl appeared to increase viability and proliferation of the cells, and CHX decreased the viability of cells.
Stannous ions caused a direct inhibition of the matrix metalloproteinases, whereas F only had an inhibitory effect when added to the zymography buffer.
Inhibition of MMPs using SnCl and SnF could play an important role in the prevention of dental erosion and caries. However, the clinical relevance of these findings needs to be proven.
牙本质和唾液中的基质金属蛋白酶 (MMPs) 可以降解胶原蛋白。二价金属是 MMPs 的已知抑制剂，但尚未对亚锡（如氯化亚锡 (SnCl) 或氟化亚锡 (SnF) 的形式）进行可能的抑制作用测试。在这项研究中，我们测试了对 MMP-2 和 MMP-9 的蛋白水解活性的抑制作用。
使用氯化钠 (NaCl)、氟化钠 (NaF) 和洗必泰 (CHX) 作为对照。使用重组人 MMP-2 和 MMP-9 进行明胶酶谱分析。SnCl、SnF、NaF、NaCl 和 CHX 分别包含在孵育缓冲液 (M1) 中或添加到重组 MMP 之前 (M2)，然后使用明胶酶谱分析 MMP。此外，使用 37%磷酸酸蚀前后，测量 SnCl、SnF 和 NaF 对人牙本质中 MMP-2 和 MMP-9 酶活性的影响。还测定了 SnCl、NaF 和 CHX 对人牙龈成纤维细胞和 L929 小鼠成纤维细胞活力的影响，以及 SnCl 和 NaF 对细胞增殖的影响。
对于 M1，观察到 SnCl 0.5%和 0.5%、SnF 0.25%和 0.12%、NaF 0.12%和 0.5%、CHX 0.012%和 0.05%的抑制浓度（w/v%）分别为 MMP-2 和 MMP-9。NaCl 没有抑制作用。对于 M2，SnCl 0.007%和 0.12%以及 SnF 0.03%和 0.5%分别抑制 MMP-2 和 MMP-9。NaF、NaCl 和 CHX 没有影响。SnCl 和 NaF 在酸蚀前涂于牙本质时，酶活性略有降低。关于用各自物质刺激细胞后的细胞活力和增殖，NaF 几乎没有影响，SnCl 似乎增加了细胞的活力和增殖，CHX 降低了细胞的活力。
亚锡离子直接抑制基质金属蛋白酶，而 F 仅在添加到明胶酶谱缓冲液中时才具有抑制作用。
使用 SnCl 和 SnF 抑制 MMPs 可能在预防牙侵蚀和龋齿方面发挥重要作用。然而，这些发现的临床相关性仍需证实。
