Objective

To evaluate the effects of different polyphenols and solvents on dentin collagen’s crosslinking interactions and biostabilization against MMPs and collagenase degradation.

Methods

Two polyphenols [proanthocyanidin (PA) and quercetin (QC)] with different water solubility were prepared as treatment solutions using ethanol (EtOH) or dimethyl sulfoxide (DMSO) as solvents. 6-um-thick dentin films were microtomed from dentin slabs of third molars. Following demineralization, films or slabs were subject to 60-s treatment (PA or QC) or no treatment (control) with subsequent extended-rinse with original solvent (EtOH or DMSO) or distilled water (DW). Collagen crosslinking interactions were assessed by FTIR. Biostability was assessed through endogenous MMPs activity via confocal laser scanning microscopy, and exogenous collagenase degradation via weight loss, hydroxyproline release and SEM. Finally, direct collagenase inactivation was also evaluated. Data were analyzed by three-way ANOVA and post-hoc tests (α = 0.05%).

Results

Distinct effects of two polyphenols and solvents on collagen crosslinking and biostabilization were observed. Higher crosslinking and biostability efficacy occurred with PA than QC (p < 0.001) that demonstrated negligible collagen interactions. With DMSO solvent, efficacy results were significantly reduced with both polyphenols (p < 0.05). DMSO-rinse further weakened interactions of PA with collagen, diminishing biostability (p < 0.05). Low biostability was detected with QC and DW-rinse, suggesting direct enzymatic inhibition due to physical presence in collagen.

Significance

Collagen crosslinking interactions and biostability depend on polyphenol chemical characteristics. Treatment-solution solvents may affect interactions between polyphenols and collagen, specifically, DMSO showed detrimental effects on collagen crosslinking and biostability and should be used with caution.

中文翻译：

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多酚和溶剂对牙本质胶原交联相互作用和生物稳定性的独特影响

客观的

评估不同多酚和溶剂对牙本质胶原蛋白交联相互作用和抗基质金属蛋白酶和胶原酶降解的生物稳定作用的影响。

方法

以乙醇（EtOH）或二甲基亚砜（DMSO）为溶剂，制备了两种水溶性不同的多酚[原花青素（PA）和槲皮素（QC）]作为处理液。从第三磨牙的牙本质薄片上切下 6 um 厚的牙本质膜。脱矿后，对薄膜或平板进行 60 秒处理（PA 或 QC）或不进行处理（对照），随后用原始溶剂（EtOH 或 DMSO）或蒸馏水（DW）进行延长冲洗。通过 FTIR 评估胶原蛋白交联相互作用。生物稳定性通过共聚焦激光扫描显微镜的内源性基质金属蛋白酶活性以及通过体重减轻、羟脯氨酸释放和 SEM 的外源性胶原酶降解进行评估。最后，还评估了直接胶原酶失活。通过三向方差分析和事后检验分析数据（α =  0.05%)。

结果

观察到两种多酚和溶剂对胶原交联和生物稳定的不同影响。PA 的交联和生物稳定性功效高于 QC (p  <  0.001)，表明胶原相互作用可忽略不计。使用 DMSO 溶剂时，两种多酚的疗效结果均显着降低 (p  <  0.05)。DMSO 冲洗进一步削弱了 PA 与胶原蛋白的相互作用，降低了生物稳定性 (p  <  0.05)。使用 QC 和 DW 冲洗检测到低生物稳定性，表明由于胶原蛋白中的物理存在而导致直接酶促抑制。

意义

胶原交联相互作用和生物稳定性取决于多酚的化学特性。处理溶液溶剂可能会影响多酚和胶原蛋白之间的相互作用，具体而言，DMSO 对胶原蛋白交联和生物稳定性有不利影响，应谨慎使用。