锶和氮改性二氧化钛/纳米羟基磷灰石复合树脂的制备和性能

doi:10.13481/j.1671-587X.20230431

摘要/Abstract

摘要：

目的制备锶和氮改性二氧化钛（Sr-N-TiO₂）/纳米羟基磷灰石（n-HA）复合树脂，并对其基础性能、抗菌性能、再矿化能力和生物安全性进行评价，阐明新型复合树脂的抗菌机制。方法 Sr-N-TiO₂与n-HA组合作为增强填料，制备复合树脂，分为不含增强填料的对照组和3个实验组，根据实验组新型复合树脂中增强填料质量分数不同又分为2.5% 增强填料组、5.0%增强填料组和7.5%增强填料组。采用傅里叶红外吸收光谱分别检测光固化60 s前后各组复合树脂的红外光谱，计算复合树脂双键转化率。使用直径 4 mm、高10 mm的模具制备各组复合树脂试件，计算光照20 s后各组复合树脂试件固化深度。每组制备3个复合树脂试件，将各组试件与稀释的变异链球菌液共培养，通过平板菌落计数法检测各组试件表面黏附细菌菌落数并计算抗菌率，采用活/死细菌染色法观察各组复合树脂试件表面活/死细菌比例及形态表现。将各组复合树脂试件于人工唾液中浸泡1、3、5和7 d，采用扫描电子显微镜（SEM）观察各组试件表面再矿化情况。小鼠纤维细胞（L-929细胞）接种于各组复合树脂浸提液中，采用CCK-8法分别于1、2和3 d时检测各组细胞相对增殖率（RGR），进行细胞毒性等级评价。结果随着复合树脂中增强填料比例的增加，新型复合树脂双键转化率和固化深度呈降低趋势，但均达到临床使用标准。与对照组比较，2.5%和5.0%增强填料组新型复合树脂双键转化率差异无统计学意义（P>0.05），7.5%增强填料组新型复合树脂双键转化率明显降低（P<0.05）。与对照组比较，各实验组新型复合树脂固化深度明显降低（P<0.05或P<0.01），表面黏附细菌菌落计数明显减少（P<0.01）。当增强填料比例达到5.0%时，新型复合树脂抗菌率大于90%。与对照组比较，各实验组新型复合树脂试件表面活细菌数随增强填料比例升高逐渐减少。SEM观察，各实验组新型复合树脂试件表面可见矿化结节，并随时间延长和增强填料比例升高而增多，处理7 d后，5.0%和7.5%增强填料组新型复合树脂试件表面几乎被矿化结节完全覆盖。生物安全性实验，各组细胞RGR均大于75%，细胞毒性等级均≤1级。结论改性后的新型复合树脂符合临床标准，并具有抗菌性能和再矿化能力，同时生物安全性较高。

关键词: 复合树脂, 龋病, 继发龋, 抗菌性, 再矿化

Abstract:

Objective To prepare the strontium and nitrogen co-doped titanium dioxide （Sr-N-TiO₂）/nano hydroxyapatite （n-HA） composite resins， to evaluate its basic properties， antibacterial properties， remineralization abilities， and biosafeties， and to clarify the antibacterial mechanism of the new type of composite resins. Methods Sr-N-TiO₂ and n-HA were mixed as the reinforcing fillers. The composite resins were prepared and were divided into control group and three experimental groups. According to the mass fractions of reinforcing fillers， the experimental groups were divided into 2.5% reinforcing filler group， 5.0% reinforcing filler group， and 7.5% reinforcing filler group. The infrared spectrogram of composite resins in various groups were detected by the Fourier-transform infrared absorption spectrogram and the double bond conversion rates were calculated before and after light curing for 60 s. The curing depths of composite resin specimens in various groups after light curing for 20 s were calculated by cylindrical mould （height=10.0 mm， diameter=4.0 mm）. Three composite resin specimens in various groups were prepared and co-cultured with the dilute Streptococcus mutans solution. The plate colony counting method was used to determine the counts of attached bacteria on surface of the specimens and the antibacterial rates were calculated； live/dead bacteria staining method was used to observe the ratio of live bacteria/dead bacteria and morphology of composite resin speciments in various groups.The composite resin specimens were immersed in the artificial saliva for 1， 3， 5， and 7 d， and the mineralizations of surface of the composite resins were observed under scanning electron microscope （SEM）. The mouse fibroblast cells（L-929 cells） were incubated with the resin extract solution，and CCK-8 method was used to detect the relative growth rates （RGR） of the cells on the 1st， 2nd， and 3rd days and the cytotoxicity levels were evaluated. Results With the increasing of reinforcing filler proportion of the composite resins， the double bond conversion rates and curing depths of new type of the composite resins showed a decreasing trend， but all reached the clinical standard. Compared with control group， the double bond conversion rates of new type of the composite resins in 2.5% reinforcing filler group and 5.0% reinforcing filler group had no significant differences （P>0.05）， and the above index in 7.5% reinforcing filler group was decreased（P<0.05）. Compared with control group， the curing depths of the composite resins in experimental groups were increased （P<0.05 or P<0.01）， the counts of attached bacteria on surface of the speciments in experimental groups were significant increased （P<0.01）. When the proportion of reinforcing fillers reached 5.0%， the antibacterical rates of new type of the composite resins were higher than 90%. Compared with control group， the counts of live bacteria on surface of the composite resins in experimental groups were decreased with the increasing of proportion of the reinforcing fillers. The SEM results show that the mineralized nodules on surface of the composite resins in experimental groups could be observed， and the number of mineralized nodules was increased with the prolongation of time and the increasing of reinforcing fillers proportion. After treated for 7 d， the surface of the composite resins in 5.0% reinforcing filler group and 7.5% reinforcing filler group were almostly covered by mineralized nodules. The biosafety experiment results showed that the RGR of cells in various groups was above 75%，and the cytotoxicity level ≤1 grade. Conclusion The modified new type of composite resins meet the clinical standards， and it has antibacterial and remineralization properties， as well as high biosafety.

Key words: Composite resin, Dental caries, Secondary caries, Antibacterial property, Remineralization

中图分类号:

R781.1

赵远航,闫琳琳,宋嘉卓,邹馨颖,赵红,刘新,陈佳文,于依岩,张志民,张红. 锶和氮改性二氧化钛/纳米羟基磷灰石复合树脂的制备和性能[J]. 吉林大学学报(医学版), 2023, 49(4): 1067-1075.

Yuanhang ZHAO,Linlin YAN,Jiazhuo SONG,Xinying ZOU,Hong ZHAO,Xin LIU,Jiawen CHEN,Yiyan YU,Zhimin ZHANG,Hong ZHANG. Preparation and properties of strontium and nitrogen co-doped titanium dioxide/nano hydroxyapatite composites resins[J]. Journal of Jilin University(Medicine Edition), 2023, 49(4): 1067-1075.

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Group	Double bonds conversion rate (n=3,η/%)	Curing depth (n=5,l/mm)
Control	55.04±1.21	2.55±0.11
2.5% reinforcing filler	54.30±2.83	2.35±0.10^*
5.0% reinforcing filler	52.27±3.80	2.29±0.13^**
7.5% reinforcing filler	48.98±1.13^*	2.18±0.07^**

Group	RGR (η/%)			Cytotoxicity grade
Group	(t/h) 24	48	72	(t/h) 24	48	72
Control	85.88	109.14	111.46	1	0	0
2.5% reinforcing filler	84.88	90.28	92.24	1	1	1
5.0% reinforcing filler	78.52	81.08	89.72	1	1	1
7.5% reinforcing filler	76.25	79.39	87.48	1	1	1

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