The requirement of zinc and calcium ions for functional MMP activity in demineralized dentin matrices.
The requirement of zinc and calcium ions for functional MMP activity in demineralized dentin matrices.
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DOI：
10.1016/j.dental.2010.07.006
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发表时间：
2010-11
期刊：
影响因子：
5
通讯作者：
Pashley, David H.
中科院分区：
文献类型：
--
作者：
Tezvergil-Mutluay, Arzu;Agee, Kelli A.;Hoshika, Tomohiro;Carrilho, Marcela;Breschi, Lorenzo;Tjaderhane, Leo;Nishitani, Yoshihiro;Carvalho, Ricardo M.;Looney, Stephen;Tay, Franklin R.;Pashley, David H.
树脂-牙本质键的逐渐降解部分是由于牙本质基质的内源性基质金属蛋白酶（MMPs）缓慢降解杂交层中的胶原原纤维。在体外耐久性研究中，储存介质的组成可能很重要，因为MMPs的最佳活性需要锌和钙。本研究评估了不同的储存介质对体外培养60天脱矿牙本质梁的基质硬度、干重损失或胶原蛋白增溶的影响。牙本质梁（1×2×6mm）在10%磷酸中完全脱矿。在基线测量干质量和弹性模量(E)（三点弯曲，15%应变）后，将梁分为5组（n=11/组），并在37°C下在含锌和钙的介质中孵育，其中包括完全介质（CM）、无钙介质、无锌介质、双锌介质或水。在孵育3、7、14、30和60天时对光束进行重新测试。用盐酸水解培养培养基，定量测定羟脯氨酸（HOP），作为MMPs对胶原增溶的指标。数据分析采用重复测量方差分析。贮藏介质和贮藏时间对E、质量损失和HOP释放均有显著影响（p<0.05）。在CM中孵育导致相对快速且显著（p<0.05）的刚度下降，质量损失增加。实验培养基的HOP含量也随培养时间的延长而增加，但显著低于推荐的对照CM培养基（p<0.05）。用于老化树脂-牙本质键的储存溶液应该是含有钙和锌的缓冲溶液。通常使用水作为老化介质可能会低估内源性牙本质MMPs的水解活性。
The progressive degradation of resin-dentin bonds is due, in part, to the slow degradation of collagen fibrils in the hybrid layer by endogenous matrix metalloproteinases (MMPs) of the dentin matrix. In in vitro durability studies, the storage medium composition might be important because the optimum activity of MMPs requires both zinc and calcium. This study evaluated the effect of different storage media on changes in matrix stiffness, loss of dry weight or solubilization of collagen from demineralized dentin beams incubated in vitro for up to 60 days. Dentin beams (1×2×6mm) were completely demineralized in 10% phosphoric acid. After baseline measurements of dry mass and elastic modulus (E) (3-point bending, 15% strain) the beams were divided into 5 groups (n=11/group) and incubated at 37°C in either media containing both zinc and calcium designated as complete medium(CM), calcium-free medium, zinc-free medium, a doubled-zinc medium or water. Beams were retested at 3, 7, 14, 30, and 60 days of incubation. The incubation media was hydrolyzed with HCl for the quantitation of hydroxyproline (HOP) as an index of solubilization of collagen by MMPs. Data were analyzed using repeated measures of ANOVA. Both the storage medium and storage time showed significant effects on E, mass loss and HOP release (p<0.05). The incubation in CM resulted in relatively rapid and significant (p<0.05) decreases in stiffness, and increasing amounts of mass loss. The HOP content of the experimental media also increased with incubation time but was significantly lower (p<0.05) than in the control CM medium, the recommended storage medium. The storage solutions used to age resin-dentin bonds should be buffered solutions that contain both calcium and zinc. The common use of water as an aging medium may underestimate the hydrolytic activity of endogenous dentin MMPs.
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