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基于羧化聚酰胺胺与镁离子协同作用构建新型牙本质再矿化体系的研究

[Study on the construction of a novel dentin remineralization system based on carboxylated polyamidoamine synergistic with magnesium ions].

作者信息

Li Q R, Long J D, Yan K Q, Huang X M, Ban G F, Xie F F, Chen W X

机构信息

Department of Endodontics, College & Hospital of Stomatology, Guangxi Medical University, Nanning 530021, China.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 2025 Aug 9;60(8):897-904. doi: 10.3760/cma.j.cn112144-20250530-00198.

DOI:10.3760/cma.j.cn112144-20250530-00198
PMID:40734402
Abstract

To explore the effect of carboxylated polyamidoamine (PAMAM-COOH) in combination with magnesium ions on the remineralization ability of amorphous calcium phosphate (ACP) in inducing remineralization of dentin collagen fibers in a 50% ethanol solution. Forty-five intact third molars extracted for impaction reasons were obtained from the College & Hospital of Stomatology, Guangxi Medical University. Two types of demineralized dentin specimens were prepared: ①Fully demineralized dentin (=30), specimens were immersed in 17% ethylenediaminetetraacetic acid (EDTA) (pH=7.4) at room temperature for 14 days with daily solution refreshment; ②Partially demineralized dentin (=15), specimens were treated with 37% phosphoric acid gel (Ultra-Etch, Ultradent) for 15 seconds followed by thorough rinsing with deionized water. Three remineralization groups were established for demineralized dentin treatment: ①Control group, 50% ethanol solution; ②ACMP group, 50% ethanol solution containing amorphous magnesium calcium phosphate (ACMP); ③PAMAM-COOH/ACMP group, 50% ethanol solution incorporating carboxylated polyamidoamine dendrimer-modified ACMP (PAMAM-COOH/ACMP). The chemical composition of remineralization solutions was analyzed by Fourier-transform infrared spectrum (FTIR). The morphology and particle size distribution of nanoparticles were characterized using transmission electron microscope (TEM). The fully demineralized dentin specimens were treated with three different remineralization solutions (37 ℃ for 7 days) respectively. The mineralization of the dentin collagen fibers surface was observed using scanning electron microscope (SEM) and the distribution of minerals inside and outside the collagen fibers was examined by using TEM. The partially demineralized dentin specimens were treated with fluorescence-labeled remineralization solutions (37 ℃ for 7 days) respectively, followed by analysis using confocal laser scanning microscopy (CLSM) to quantitatively evaluate the penetration depth of the mineralization agents. FTIR analysis confirmed the presence of characteristic absorption peaks corresponding to phosphate (PO) groups, carbon-nitrogen bonds, and amide linkages in the PAMAM-COOH/ACMP nanocomposite. TEM observed that the PAMAM-COOH/ACMP nanoparticles exhibited an average particle size of (36.85±8.02) nm in an amorphous state. SEM observation indicates continuous mineral deposition on dentin collagen fibers in the PAMAM-COOH/ACMP group, while no mineral deposition in the control group and only minimal deposition in the ACMP group. TEM showed no mineral deposition inside or outside the collagen fibers in the control group, only external mineral deposition in the ACMP group, and high-density mineral deposition both inside and outside the fibers in the PAMAM-COOH/ACMP group. CLSM analysis revealed a statistically significant difference (<0.05) in the depth of mineralized substances entering dentin tubules between ACMP group and PAMAM-COOH/ACMP group. The remineralization system of 50% ethanol solution incorporating PAMAM-COOH/ACMP successfully achieved the internal and external mineralization of demineralized dentin collagen fibers.

摘要

为探究羧基化聚酰胺-胺(PAMAM-COOH)与镁离子联合作用对无定形磷酸钙(ACP)在50%乙醇溶液中诱导牙本质胶原纤维再矿化能力的影响。从广西医科大学附属口腔医院获取因阻生原因拔除的45颗完整第三磨牙。制备两种类型的脱矿牙本质标本:①完全脱矿牙本质(n=30),标本在室温下于17%乙二胺四乙酸(EDTA)(pH=7.4)中浸泡14天,每天更换溶液;②部分脱矿牙本质(n=15),标本用37%磷酸凝胶(Ultra-Etch,Ultradent)处理15秒,然后用去离子水彻底冲洗。为脱矿牙本质处理建立三个再矿化组:①对照组,50%乙醇溶液;②ACMP组,含无定形镁钙磷酸盐(ACMP)的50%乙醇溶液;③PAMAM-COOH/ACMP组,含羧基化聚酰胺-胺树枝状大分子修饰的ACMP(PAMAM-COOH/ACMP)的50%乙醇溶液。通过傅里叶变换红外光谱(FTIR)分析再矿化溶液的化学成分。使用透射电子显微镜(TEM)表征纳米颗粒的形态和粒径分布。完全脱矿牙本质标本分别用三种不同的再矿化溶液(37℃,7天)处理。使用扫描电子显微镜(SEM)观察牙本质胶原纤维表面的矿化情况,并用TEM检查胶原纤维内外矿物质的分布。部分脱矿牙本质标本分别用荧光标记的再矿化溶液(37℃,7天)处理,然后使用共聚焦激光扫描显微镜(CLSM)进行分析,以定量评估矿化剂的渗透深度。FTIR分析证实PAMAM-COOH/ACMP纳米复合材料中存在对应于磷酸根(PO)基团、碳氮键和酰胺键的特征吸收峰。TEM观察到PAMAM-COOH/ACMP纳米颗粒呈无定形态,平均粒径为(36.85±8.02)nm。SEM观察表明,PAMAM-COOH/ACMP组牙本质胶原纤维上有连续的矿物质沉积,而对照组无矿物质沉积,ACMP组仅有少量沉积。TEM显示,对照组胶原纤维内外均无矿物质沉积,ACMP组仅在外部有矿物质沉积,而PAMAM-COOH/ACMP组纤维内外均有高密度矿物质沉积。CLSM分析显示,ACMP组和PAMAM-COOH/ACMP组之间矿物质进入牙本质小管的深度存在统计学显著差异(<0.05)。含PAMAM-COOH/ACMP的50%乙醇溶液再矿化系统成功实现了脱矿牙本质胶原纤维的内外矿化。

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