氧化海藻酸钠预处理提高树脂牙本质粘接的耐久性

doi:10.3877/cma.j.issn.1674-1366.2018.06.001

目的

研究氧化海藻酸钠（ADA）改性脱矿牙本质胶原的交联度和显微形貌，及其对牙本质基质金属蛋白酶（MMP）的抑制作用和树脂牙本质粘接强度的影响。

方法

制备ADA，进行傅里叶红外线光谱分析（FTIR）。采用0.5%、1%、2%、3%和4%的ADA处理脱矿牙本质粉末（DDP）1、2、30 min，5%戊二醛溶液作为阳性对照，茚三酮比色法检测胶原交联度。制备（1.0 ± 0.1）mm厚牙本质片，35%磷酸溶液酸蚀后ADA处理1 min，保持表面干燥或湿润，场发射扫描电镜（FESEM）观察其显微形貌。Sensolyte Generic MMP assay kit试剂盒检测不同浓度ADA对MMP-9的抑制作用。制备树脂牙本质粘接样本，检测ADA预处理试件冷热循环前后的微拉伸强度。使用SPSS 22.0对交联度实验进行多因素方差分析和Tukey检验，MMP-9抑制实验进行秩和检验和Bonferroni检验，微拉伸实验进行单因素方差分析和LSD检验。

结果

FTIR显示海藻酸钠氧化后生成醛基形成ADA。DDP经ADA处理后，胶原交联度随处理浓度及处理时间呈依赖性增加，不同浓度组间（F = 1329.423，P<0.001）及不同时间组间（F = 1142.93，P<0.001）差异均有统计学意义。35%磷酸溶液脱矿牙本质表面经处理后，除阴性对照和0.5% ADA干燥处理组胶原纤维塌陷外，其余组均保持蓬松状态。0.5% ~ 4% ADA对MMP-9的抑制程度为93.5% ~ 100%，不同浓度组间差异有统计学意义（F = 13.786，P<0.001）。ADA预处理后，2% ADA组粘接样本冷热循环前后的粘接强度最高，分别为（28.2 ± 4.2）、（18.3 ± 3.7）MPa，差异有统计学意义（F_前 = 5.544，P_前<0.001；F_后 = 5.181，P_后<0.001）MPa。

结论

2% ADA处理脱矿牙本质可提高Ⅰ型胶原纤维的交联度，使胶原纤维网保持蓬松状态，同时能抑制MMP-9活性，提高树脂牙本质粘接即刻及老化后的微拉伸强度，有利于改善树脂牙本质的粘接耐久性。

关键词: 藻酸盐, 交联试剂，天然, 树脂粘固，牙本质, 氧化海藻酸钠

Objective

To investigate the effect of alginate dialdehyde (ADA) on demineralized dentin substrate, in its cross-linking degree, micromorphology, inhibition ability and resin-dentin bonding strength.

Methods

ADA was prepared, and analyzed by Fourier Transform infrared spectroscopy (FTIR) . Demineralized dentin powder were treated by 0.5%, 1%, 2%, 3%, 4% ADA for 1, 2, 30 min, respectively, and examined by Nihydri colorimetry for the cross-linking degrees, 5% Glutaraldehyde was used as positive control (1.0 ± 0.1) mm dentin discs were etched, rinsed and pretreated with ADA successively, and kept dry or moist for field emission scanning electron microscope (FESEM) . The inhibition of ADA on MMP-9 was analyzed by Sensolyte Generic MMP assay kit. Resin-dentin bonding specimens were prepared for micro-tensile bond strength evaluation (MTBS) , before and after thermolcycling. Data were analyzed by Multi-way ANOVA, Kruskal-Wallis test or One-way ANOVA by SPSS 22.0 software package.

Results

FTIR analysis confirmed the characteristic peak of aldehyde group in ADA. The cross-linking degree of ADA-treated DDP was exhibited concentration- and time-dependent and from which significant differences were found among different ADA concentration (F = 1329.423, P<0.001) and different time (F = 1142.93, P<0.001) . The FESEM result confirmed that the demineralized dentin collagen was in a homogenous and regular arrangement after ADA or GD treatment, except 0.5% ADA dry group. Over 93.5% of the MMP-9 was inhibited by 0.5% ~ 4% ADA (F = 13.786, P<0.001) . The 2% ADA group exhibited highest MTBS before [ (28.2 ± 4.2) MPa] (F = 5.544, P<0.001) and after [ (18.3 ± 3.7) MPa] (F = 5.181, P<0.001) thermolcycling.

Conclusions

2% ADA could increase cross-linking degree of demineralized dentin substrate, keep collagen fiber from collapse, and inhibit the activity of MMP-9, thus increase the durability of the dentin adhensive.

Key words: Alginates, Cross-linking reagents, natural, Resin-bonded, dentin, Alginate dialdehyde

图1 海藻酸钠氧化前后的红外光谱图

表1 不同浓度氧化海藻酸钠预处理后胶原交联度（%， ± s）

组别	1 min	2 min	30 min
5% GD组	55.4 ± 2.8^Aa	84.5 ± 0.9^Ba	97.0 ± 1.0^Ca
0.5% ADA组	2.5 ± 0.9^Ab	15.6 ± 2.9^Bb	21.6 ± 3.7^Cb
1% ADA组	8.2 ± 2.9^Ac	21.5 ± 2.2^Bc	30.6 ± 4.6^Cc
2% ADA组	19.4 ± 3.0^Ad	21.6 ± 2.9^Ac	36.3 ± 3.7^Bd
3% ADA组	21.5 ± 2.6^Ad	24.0 ± 3.9^Ac	38.7 ± 5.4^Bd
4% ADA组	22.1 ± 2.9^Ad	24.0 ± 3.5^Ac	46.4 ± 5.4^Be

表1 不同浓度氧化海藻酸钠预处理后胶原交联度（%， ± s）

组别	1 min	2 min	30 min
5% GD组	55.4 ± 2.8^Aa	84.5 ± 0.9^Ba	97.0 ± 1.0^Ca
0.5% ADA组	2.5 ± 0.9^Ab	15.6 ± 2.9^Bb	21.6 ± 3.7^Cb
1% ADA组	8.2 ± 2.9^Ac	21.5 ± 2.2^Bc	30.6 ± 4.6^Cc
2% ADA组	19.4 ± 3.0^Ad	21.6 ± 2.9^Ac	36.3 ± 3.7^Bd
3% ADA组	21.5 ± 2.6^Ad	24.0 ± 3.9^Ac	38.7 ± 5.4^Bd
4% ADA组	22.1 ± 2.9^Ad	24.0 ± 3.5^Ac	46.4 ± 5.4^Be

图2 不同方式预处理脱矿牙本质表面后的显微形貌

图3 不同浓度氧化海藻酸钠（ADA）对基质金属蛋白酶（MMP）9活性的抑制程度

表2 氧化海藻酸钠（ADA）预处理粘接样本冷热循环前后的粘接强度（MPa， ± s）

组别	例数	未经冷热循环	冷热循环后
阴性对照组	30	27.2 ± 3.3^A	14.4 ± 3.4^a
5% GD组	30	29.4 ± 3.1^B	17.9 ± 4.2^bc
0.5% ADA组	30	27.4 ± 4.0^A	16.6 ± 3.4^bc
1% ADA组	30	27.1 ± 3.7^A	18.0 ± 3.0^bc
2% ADA组	30	28.2 ± 4.2^AB	18.3 ± 3.7^b
3% ADA组	30	27.1 ± 3.5^A	16.3 ± 3.4^c
4% ADA组	30	24.2 ± 4.1^C	-

表2 氧化海藻酸钠（ADA）预处理粘接样本冷热循环前后的粘接强度（MPa， ± s）

组别	例数	未经冷热循环	冷热循环后
阴性对照组	30	27.2 ± 3.3^A	14.4 ± 3.4^a
5% GD组	30	29.4 ± 3.1^B	17.9 ± 4.2^bc
0.5% ADA组	30	27.4 ± 4.0^A	16.6 ± 3.4^bc
1% ADA组	30	27.1 ± 3.7^A	18.0 ± 3.0^bc
2% ADA组	30	28.2 ± 4.2^AB	18.3 ± 3.7^b
3% ADA组	30	27.1 ± 3.5^A	16.3 ± 3.4^c
4% ADA组	30	24.2 ± 4.1^C	-

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Preliminary study on resin-dentin bond durability by alginate dialdehyde preconditioning

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Preliminary study on resin-dentin bond durability by alginate dialdehyde preconditioning