载氟聚碳酸丙烯酯牙贴片再矿化性能及其对小鼠成纤维L929细胞的毒性作用

doi:10.13481/j.1671-587X.20220608

摘要/Abstract

摘要：

目的制备以聚碳酸丙烯酯（PPC）和氟化钠（NaF）为基础的新型PPC/NaF牙贴片，探讨其再矿化性能和体外细胞毒性，阐明不同氟含量的含氟贴片促进脱矿釉质再矿化效果，为其临床应用提供参考。方法以PPC为载体搭载不同质量分数（0、0.5%、2.5%和5.0%）NaF，采用熔融共混法制备PPC/NaF牙贴片。将离体牙制备成釉质块试件，并根据牙贴片中氟含量的不同随机分为PPC组、0.5%PPC/NaF组、2.5%PPC/NaF组和5.0%PPC/NaF组，每组各10个样本。采用扫描电子显微镜（SEM）观察各组试件釉质表面形态表现，显微硬度仪测定脱矿前后和再矿化后各组试件釉质表面显微硬度，并计算显微硬度恢复百分比（SMHR）。将小鼠成纤维L929细胞与PPC/NaF牙贴片浸提液进行共培养，CCK-8实验检测各组小鼠成纤维L929细胞的相对增殖率（RGR），并评估其细胞毒性等级。结果脱矿后各组釉质试件的表面显微硬度均明显低于脱矿前（P<0.05）。经不同氟含量PPC/NaF牙贴片处理后，各组釉质试件的再矿化后表面显微硬度均明显高于再矿化前（P<0.05），但仍低于脱矿前（P<0.05），各组釉质试件的SMHR由高到低依次为5.0%PPC/NaF组、2.5%PPC/NaF组、0.5%PPC/NaF组和PPC组。SEM观察，PPC组釉质试件表面孔隙数量仍较多，表面可见不规则的颗粒状沉积物；0.5%PPC/NaF组釉质试件表面孔隙明显减少，但表面仍可见明显釉质塌陷且晶体沉积疏松，不均匀；2.5%PPC/NaF组釉质试件表面几乎不见孔隙，表面较为规则并覆盖大量针状晶体，排列密集紧凑；5.0%PPC/NaF组釉质试件表面被大量针状或不规则颗粒状的晶体覆盖，有些区域可见层状沉积物。与PPC组比较，5.0%PPC/NaF组小鼠成纤维L929细胞RGR明显降低（P<0.05），各组小鼠成纤维L929细胞采用不同氟含量的PPC/NaF牙贴片处理后的RGR均大于75%，细胞毒性等级为0级或1级。结论 PPC/NaF牙贴片能够有效促进脱矿釉质发生再矿化，无细胞毒性；随着氟含量的增加，其再矿化作用也随之增强。

关键词: 聚碳酸丙烯酯, 氟化物, 牙贴片, 再矿化, 细胞增殖率

Abstract:

Objective To prepare the novel fluoride loaded poly （prolylene carbonate）（PPC）/sodium fluoride （NaF） dental patch based on PPC and explore its remineralization property and in vitro cytotoxicity， and to clarify its effect of fluorine-containing patches with different concentrations of fluoride in promoting the remineralization of demineralized enamel， and to provide the reference for its clinical application. Methods The PPC/NaF dental patch with different mass fractions of sodium fluoride （0， 0.5%， 2.5%，and 5.0%） were prepared by melt-blending method. The enamel blocks were prepared with extracorporeal tooth and randomly divided into PPC group， 0.5%PPC/NaF group， 2.5%PPC/NaF group， and 5.0%PPC/NaF group according to different contents of fluoride in the dental patches， and there were 10 samples in each group. The morphology of surface of enamel specimens was observed under scanning electron microscope（SEM），the microhardness of the enamel surface before and after demineralization and after remineralization was measured by microhardness tester， and the microhardness recovery percentage （SMHR） was calculated. The mouse fibroblast L929 cells were co-cultured with the PPC/NaF dental patch extract，and CCK-8 test was used to detect the relative proliferation rate （RGR） of the fibroblast L929 cells of the mice in various groups， and its cytotoxicity level was evaluated. Results The microhardness of the enamel specimens in various groups after demineralization was significantly lower than that before demineralization （P<0.05）. After being treated with PPC/NaF tooth patches with different contents of sodium fluoride， the microhardness of the enamel specimens in various groups was significantly higher than that before remineralization（P<0.05）， but still lower than that before demineralization （P<0.05）. The SMHR of enamel specimens in each group from high to low was 5.0% PPC/NaF group， 2.5% PPC/NaF group， 0.5% PPC/NaF group， and PPC group. The SEM results showed that the number of pores on the surface of the enamel specimens in PPC group was still large， and the irregular granular sediments were seen； in 0.5%PPC/NaF group， the number of pores on the enamel surface was decreased significantly，and the obvious enamel collapse was still seen on the surface and mineral crystal deposition was loose and uneven； in 2.5% PPC/NaF group， there were almost no pores on the enamel surface，and the enamel surface was regular and covered with a large number of needle-like crystals， which were densely arranged； in 5.0% PPC/NaF group， the enamel surface was covered with a large number of needle-like or irregular granular crystals and layered sediments were seen in some areas. Compared with PPC group， the RGR of the fibroblast L929 cells of the mice in 5.0% PPC/NaF group was decreased significantly （P<0.05）. The RGR of the fibroblast L929 cells after treated with PPC/NaF tooth patches with different contents of fluoride in various groups was greater than 75%， and the cytotoxicity grade was 0 or 1. Conclusion PPC/NaF dental patch can effectively promote the demineralization of the demineralized enamel without cytotoxicity；with the increasing of fluorine concentration， its remineralization is also enhanced.

Key words: Polypropylene carbonate, Fluoride, Dental patch, Remineralization, Cell proliferation rate

中图分类号:

R781.1

陈星竹,于明岳,刘双,李佳凝,谢尊玄,刘金瑶,刘玉艳. 载氟聚碳酸丙烯酯牙贴片再矿化性能及其对小鼠成纤维L929细胞的毒性作用[J]. 吉林大学学报(医学版), 2022, 48(6): 1429-1436.

Xingzhu CHEN,Mingyue YU,Shuang LIU,Jianing LI,Zunxuan XIE,Jinyao LIU,Yuyan LIU. Remineralization property of fluoride loaded poly (propylene carbonate) dental patch and its cytotoxicity on fibroblast L929 cells of mice[J]. Journal of Jilin University(Medicine Edition), 2022, 48(6): 1429-1436.

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Group	SMH₀ (kg·mm^－2)	SMH₁ (kg·mm^－2)	SMH₂ (kg·mm^－2)	SMHR (η/%)
PPC	353.75±13.41	234.79±20.90^*	246.07±19.01^*△	9.44±3.22
0.5%PPC/NaF	349.75±14.89	230.42±19.20^*	268.83±16.37^*△	36.55±11.88^#
2.5%PPC/NaF	349.17±12.50	236.63±14.32^*	299.13±18.47^*△	56.60±16.78^#○
5.0%PPC/NaF	356.13±15.85	232.46±18.11^*	328.29±20.71^*△	77.11±11.49^#○□
SMH₀：SMH at baseline；SMH₁：SMH after demineralization； SMH₂：SMH after remineralization.^*P<0.05 compared with SMH₀；^△P<0.05 compared with SMH₁；^#P<0.05 compared with PPC group；^○P<0.05 compared with 0.5%PPC/NaF group； ^□P<0.05 compared with 2.5%PPC/NaF group.

Group	RGR(η/%)	Cytotoxicity grade
Blank control	100	0
PPC	101.50±5.49	0
0.5%PPC/NaF	98.81±6.69	1
2.5%PPC/NaF	90.50±4.48	1
5.0%PPC/NaF	78.27±2.03^*	1

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