Effect of chlorhexidine-loaded poly(amido amine) dendrimer on matrix metalloproteinase activities and remineralization in etched human dentin in vitro*
Effect of chlorhexidine-loaded poly(amido amine) dendrimer on matrix metalloproteinase activities and remineralization in etched human dentin in vitro*
复制标题
负载氯己定的聚酰胺胺树枝状聚合物对体外蚀刻的人牙本质基质金属蛋白酶活性和再矿化的影响
DOI：
10.1016/j.jmbbm.2021.104625
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发表时间：
2021-06-12
影响因子：
3.9
通讯作者：
Yang, Deqin
中科院分区：
文献类型：
--
作者：
Chen, Liang;Chen, Wang;Yang, Deqin
为研究负载氯己定（CHX）的端羧基聚酰胺胺树状大分子（CHX-PAMAM-COOH）对人牙本质基质金属蛋白酶（MMP）活性及矿化的影响，制备了CHX-PAMAM-COOH树状大分子，并采用傅里叶变换红外光谱对其进行了表征。使用MMP活性测定试剂盒测量CHX、PAMAM-COOH和CHX-PAMAM-COOH对可溶性重组人基质金属蛋白酶（rhMMP-2）和牙本质结合的内源性MMP活性的抑制作用。采用原位酶谱法检测CHX、PAMAM-COOH和CHX-PAMAM-COOH预处理牙本质中明胶酶活性。用场发射扫描电镜（SEM）和能谱仪（EDS）观察CHX、PAMAMCOOH和CHX-PAMAM-COOH预处理的牙本质在人工唾液中孵育14 d后的再矿化情况。rhMMP-2活性测定结果显示，CHX-PAMAM-COOH组和CHX组的MMP-2活性显著降低至5.58 ± 0.85%，而PAMAM-COOH组则显著增加至213.38 ± 0.11%（P <0.05）。总MMP活性和原位酶谱结果显示，CHXPAMAM-COOH组和CHX组牙本质中内源性明胶酶活性显著降低。扫描电镜和能谱分析结果表明，PAMAM-COOH组和CHX-PAMAM-COOH组牙本质表面形成棒状晶体，Ca/P值分别为1.73和1.71。结论：CHX-PAMAM-COOH能抑制牙本质中内源性MMPs的结合，同时诱导酸蚀牙本质的再矿化。然而，重要的是要注意，PAMAM树枝状聚合物的催化作用可能对MMP活性具有不期望的兴奋作用，如果在口腔环境中单独使用PAMAM树枝状聚合物，则不能忽略这一点。
To investigate the effect of chlorhexidine (CHX)-loaded carboxyl-terminated poly (amido amine) dendrimer (CHX-PAMAM-COOH) on matrix metalloproteinase (MMP) activities and remineralization in human dentin, CHX-PAMAM-COOH was prepared and characterized by Fourier-transform infrared spectroscopy. The inhibitory effects of CHX, PAMAM-COOH, and CHX-PAMAM-COOH on soluble recombinant human matrix metalloproteinase (rhMMP-2) and dentin-bound endogenous MMP activity were measured using an MMP Activity Assay Kit. In situ zymography was performed to evaluate the gelatinase activity in dentin pretreated with CHX, PAMAM-COOH, and CHX-PAMAM-COOH. The remineralization of etched dentin pretreated with CHX, PAMAMCOOH, and CHX-PAMAM-COOH was evaluated by field emission-scanning electron microscopy (SEM) and energy disperse spectroscopy (EDS) after incubation in artificial saliva for 14 days. The results of the rhMMP-2 activity assay showed that the MMP-2 activity in the CHX-PAMAM-COOH group and the CHX group decreased significantly to 5.58 +/- 0.85% (P < 0.05) and 4.86 +/- 1.12% (P < 0.05), respectively, but that in the PAMAM-COOH group increased significantly to 213.38 +/- 0.11% (P < 0.05). The results of total MMP activity and in situ zymography showed a significant reduction in endogenous gelatinase activity in dentin in the CHXPAMAM-COOH group and the CHX group. The SEM and EDS results showed that rod-like crystals were formed on the etched dentin surface in the PAMAM-COOH group and the CHX-PAMAM-COOH group, and their Ca/P ratios were 1.73 and 1.71, respectively. In conclusion, CHX-PAMAM-COOH can inhibit dentin-bound endogenous MMPs and induce remineralization in etched dentin simultaneously. However, it is important to note that the catalytic role of PAMAM dendrimers may have an undesired excitatory effect on MMP activity, which cannot be ignored if PAMAM dendrimers were used alone in the oral environment.