Effect of biomimetic mineralization on enamel and dentin: A Raman and EDX analysis

ObjectiveTo investigate the effect of an experimental biomimetic mineralization kit (BIMIN) on the chemical composition and crystallinity of caries-free enamel and dentin samples in vitro.MethodsEnamel and dentin samples from 20 human teeth (10 for enamel; 10 for dentin) were divided into a control group without treatment and test samples with BIMIN treatment. Quantitative analysis of tissue penetration of fluoride, phosphate, and calcium was performed using energy-dispersive X-ray spectroscopy (EDX). Mineralization depth was measured by Raman spectroscopy probing the symmetric valence vibration near 960 cm⁻¹ as a marker for crystallinity. EDX data was statistically analyzed using a paired t-test and Raman data was analyzed using the Student’s t-test.ResultsEDX analysis demonstrated a penetration depth of fluoride of 4.10 ± 3.32 μm in enamel and 4.31 ± 2.67 μm in dentin. Calcium infiltrated into enamel 2.65 ± 0.64 μm and into dentin 5.58 ± 1.63 μm, while the penetration depths for phosphate were 4.83 ± 2.81 μm for enamel and 6.75 ± 3.25 μm for dentin. Further, up to 25 μm of a newly mineralized enamel-like layer was observed on the surface of the samples. Raman concentration curves demonstrated an increased degree of mineralization up to 5–10 μm into the dentin and enamel samples.SignificanceBiomimetic mineralization of enamel and dentin samples resulted in an increase of mineralization and a penetration of fluoride into enamel and dentin.     

Effect of air-abrasion pre-treatment with bioactive glass 45S5 on enamel surface loss after erosion/abrasion challenge

ObjectivesThe aim of this in vitro study was to evaluate the effect of air-abrasion surface pre-treatment with bioactive glass (BAG) 45S5 on enamel surface loss after erosion/abrasion challenge.MethodsTwenty-four sound bovine incisors were used for this study. Four experimental groups (n = 12) were assigned as follows: Group 1 was the negative control group, Group 2 specimens were treated with a SnF₂ gel (positive control group), Group 3 specimens were air-abraded with BAG 45S5 (ProSylc) and Group 4 received both treatments. The specimens were submitted to erosion/abrasion challenge using a common soft drink. Enamel surface loss was evaluated using an optical profilometer. Additionally, surface roughness (VSI method) and surface microhardness (Vickers method) changes were evaluated, as well as SEM and EDS analyses were performed on enamel surface. The data were statistically analyzed using one-way ANOVA and Tukey’s post-hoc test at a level of significance a = 0.05.ResultsSurface pre-treatment with BAG 45S5 reduced surface loss in comparison with negative control group (p < 0.001), which exhibited the highest surface loss of the experimental groups (p < 0.05). The positive control group (SnF₂ treatment) and Group 4 specimens, which received both air-abrasion pre-treatment and SnF₂ treatment, presented the lowest surface loss (p < 0.05), but did not show significant differences to each other (p = 0.65).SignificanceAir-abrasion pre-treatment with BAG 45S5 may be beneficial as an in-office preventive method for the limitation of enamel erosive wear induced by excessive consumption of soft drinks. The clinical significance of the results regarding this preventive method should be confirmed by clinical studies.     

釉基质蛋白对人牙髓细胞成牙本质分化的影响

目的 探讨釉基质蛋白对人牙髓细胞(human dental pulp cells,HDPC)成牙本质分化的影响,为牙本质的组织工程学研究提供有效的生物活性物质.方法 将HDPC接种于6孔板(2×105个/孔)后分为5组,分别加入含1、10、100 mg/L釉基质蛋白(enamel matrix proteins,EMP)(分别为EMP 1、10、100 mg/L组)、10-8 mol/L地塞米松及100 mg/L抗坏血酸(dexamethasone and ascorbic acid,Dex-AA组)、单纯基础培养液(对照组).于培养1、5、10 d分别用碱性磷酸酶(alkaline phosphatase,ALP)试剂盒检测ALP活性,实时荧光定量反转录聚合酶链反应技术检测成牙本质相关基因牙本质基质蛋白1(dentine matrix protein-1,DMP-1)及牙本质涎磷蛋白(dentine sialophosphoprotein,DSPP)的表达,应用2-△△CT方法得出具体数值并进行单因素方差分析与Post Hoe 检验,用茜素红染色检测并定量分析矿化情况.结果 各组ALP水平呈时间依赖性上调,培养1d后各组ALP活性与对照组相比差异均无统计学意义;5 d后EMP 10、100 mg/L组及Dex-AA组ALP活性显著增高,分别达到7.573±0.267、6.119±0.502、5.846±0.096,与对照组相比差异均有统计学意义(P＜0.05);10 d后10 mg/L EMP组及Dex-AA组ALP活性增高更显著,分别达21.035±0.149、13.223±0.797,与对照组(5.825±0.404)相比差异均有统计学意义(P＜0.01).培养1d后各组DMP-1及DSPP mRNA水平均较对照组显著升高,差异均有统计学意义(P＜0.01);培养10 d后10 mg/L EMP组DMP-1和DSPP表达水平均显著增高,分别达到14.791±0.164、12.238±0.421.培养10 d后各组均有矿化,10 mg/L EMP组钙离子浓度[(191.8±2.0) μmol/L]显著高于对照组[(81.1±8.1)μmol/L],差异有统计学意义(P＜0.01).结论 适当浓度的EMP对HDPC的成牙本质分化有一定的促进作用.     

四种正畸粘接剂对金属托槽粘接效果的评价

目的比较4种正畸粘接剂对金属托槽的粘接效果。方法选择正畸临床44例患者,792颗牙,随机分为4组,常规牙面处理,分别按照说明书要求用京津釉质粘接剂、GC树脂改良型玻璃离子粘接剂、3M化学固化型树脂粘接剂和3M光固化树脂粘接剂粘接金属托槽,然后使用MBT矫治技术进行综合性正畸治疗,记录托槽脱落情况。12个月后,评估每组牙的牙釉质脱矿情况,并对所有数据进行统计学分析。结果 3M光固化型树脂粘接剂组托槽的脱落率低于京津釉质粘接剂组,差异有显著性(P<0.05),而其余各组间托槽脱落率的比较差异均无统计学意义(P>0.05)。京津釉质粘接剂组造成釉质脱矿程度在4组中最高,而GC树脂改良型玻璃离子粘接剂的釉质脱矿程度最低,其差异均有统计学意义(P<0.05)。结论京津釉质粘接剂相对远期粘接效果较差;GC树脂改良型玻璃离子粘接剂能够通过释放氟离子有效降低釉质脱矿的发生,是正畸治疗中较为理想的粘接材料,具有良好的临床应用前景。     

Non-invasive imaging of the crystalline structure within a human tooth

The internal crystalline structure of a human molar tooth has been non-destructively imaged in cross-section using X-ray diffraction computed tomography. Diffraction signals from high-energy X-rays which have large attenuation lengths for hard biomaterials have been collected in a transmission geometry. Coupling this with a computed tomography data acquisition and mathematically reconstructing their spatial origins, diffraction patterns from every voxel within the tooth can be obtained. Using this method we have observed the spatial variations of some key material parameters including nanocrystallite size, organic content, lattice parameters, crystallographic preferred orientation and degree of orientation. We have also made a link between the spatial variations of the unit cell lattice parameters and the chemical make-up of the tooth. In addition, we have determined how the onset of tooth decay occurs through clear amorphization of the hydroxyapatite crystal, and we have been able to map the extent of decay within the tooth. The described method has strong prospects for non-destructive probing of mineralized biomaterials.     

Developmental enamel defects in the primary dentition: aetiology and clinical management

Developmental enamel defects, presenting as enamel hypoplasia or opacities are caused by damage or disruption to the developing enamel organ as a result of inherited and acquired systemic conditions. The high prevalence of these defects in the primary dentition demonstrates the vulnerability of the teeth to changes in the pre‐ and postnatal environment. The presence of enamel hypoplasia increases the risk of primary teeth to early childhood caries and tooth wear as the defective enamel is thinner, more plaque retentive and less resistant to dissolution in acid compared to normal enamel. The purpose of this paper was to critically review the aetiology and clinical complications of developmental enamel defects in the primary dentition and propose recommendations for the clinical management of affected teeth.     

Measuring enamel erosion: A comparative study of contact profilometry,non-contact profilometry and confocal laser scanning microscopy

Objectives

To compare three instruments for their ability to quantify enamel loss after acid erosion.

Methods

6 randomized parallel groups of bovine enamel samples were subjected to citric acid (higher acidity) or orange juice (lower acidity) erosion and remineralisation in a cycling model. Two protected shoulders were created on each of the samples using tape, to serve as reference for analysis. The time of exposure to each acid was varied, along with presence or absence of agitation. After treatment, samples were measured on 3 instruments capable of measuring step height: a contact profilometer (CP); a non-contact profilometer (NCP); and a confocal laser scanning microscope (CLSM) by three different examiners. Additionally, 3D (volume) step height was also measured using the CLSM.

Results

Increasing acid concentration and exposure time resulted in greater erosion, as did agitation of samples while in acid solution. All instruments/methods identified the same statistically significant (p < 0.05) pair-wise differences between the treatments groups. Further, all four methods exhibited strong agreement (Intra-class correlation ≥ 0.96) in erosion level and were highly correlated, with correlations of 0.99 or higher in all cases.

Significance

All instruments/methods used in this study produced very similar conclusions with regard to ranking of enamel loss, with data showing very high agreement between instruments. All instruments were found to be equally suited to the measurement of enamel erosion.     

Characterisation of developmentally hypomineralised human enamel

Objectives

To investigate and clarify physical and chemical properties of enamel affected by molar incisor hypomineralisation (MIH).

Methods

A series of in vitro studies were performed on extracted molars affected by MIH and sound teeth for controls. Tooth sections underwent Vickers microhardness testing before lapping and subsequent transverse microradiographic analysis and examination under polarised light microscopy. Carbonate content was determined by CO₂ release from acid digestion. Unprepared and fractured surfaces were examined under scanning electron microscopy.

Results

MIH-affected molars demonstrated a severe degree of hypomineralisation with an average mineral content of only 58.8% vol% mineral. Vickers microhardness was significantly reduced in MIH compared with controls (1.8 ± 1.1 v 4.4 ± 1.0 GPa, p < 0.05) and polarised light microscopy revealed the bulk of MIH lesions had a porosity of ≤5% but also substantial areas of ≥10% and smaller areas exceeding 25% porosity. A surface layer was frequently observed on both intact and broken-down lesions and cervical regions of MIH teeth were typically spared. Carbonate content of MIH enamel was higher than control samples (6.6 ± 2.1 v 4.4 ± 1.1 wt%, p < 0.05). Scanning electron microscopy showed that both the enamel rod and surface ultrastructure were defective. Clinical characteristics did not consistently correlate with all properties.

Conclusions

The properties of MIH-affected enamel significantly differ from those of normal enamel and were highly variable, however some common characteristics were observed. Implications for aetiology and clinical management are discussed.     

GC护牙素在固定正畸中预防釉质脱矿的临床研究

目的观察固定正畸矫治中GC护牙素预防牙釉质脱矿的临床效果。方法选择固定矫治器治疗的患者64名,随机分为2组,护牙素组32例,657颗牙;对照组32例,678颗牙。矫治结束后对牙面脱矿情况进行临床观察,用SPSS13.0统计软件进行数据处理分析。结果矫治前二组患者被观察牙齿牙釉质脱矿率和脱矿程度均无明显差异(P>0.05)。矫治后护牙素组牙釉质脱矿率为5.78%,对照组为15.78%。护牙素处理组牙釉质脱矿率和脱矿程度明显低于对照组,有显著性差异(P<0.01)。结论在固定正畸治疗中配合使用GC护牙素处理牙面,可预防牙釉质脱矿的发生。     

Efficacy of the stannous ion and a biopolymer in toothpastes on enamel erosion/abrasion

Objectives

Sn²⁺ has promising erosion-inhibiting properties in solutions, but little is known about respective effects in toothpastes. In addition, biopolymers might have protecting potential. Aim of this study was to investigate the effects of Sn²⁺ in toothpastes and of a biopolymer (chitosan) added to a Sn²⁺ formulation on erosion/abrasion.

Methods

Enamel samples were subjected to cyclic erosion procedures (10 days; 0.50% citric acid, pH 2.5; 6× 2 min/day), and brushing (2× 15 s/day, load 200 g) during immersion in slurries (2 min). The toothpastes were NaF formulations (NaF/1, NaF/2, NaF/3) and Sn²⁺ formulations (NaF/SnCl₂, AmF/SnF₂, AmF/NaF/SnCl₂) and AmF/NaF/SnCl₂ + 0.5% chitosan.Declared concentrations of active ingredients in toothpastes were 1400–1450 μg/g F⁻ and 3280–3500 μg/g Sn²⁺. Negative controls were erosion only and placebo, positive control was a SnF₂ gel. Tissue loss was quantified profilometrically, Sn on enamel surfaces was measured by energy dispersive X-ray spectroscopy.

Results

Loss values (μm) for erosion only and placebo were 14.4 ± 4.5 and 20.2 ± 3.8, respectively, and 4.6 ± 1.9 for the positive control (p ≤ 0.001 each compared to erosion only). The other loss values were: NaF/1 16.5 ± 3.0, NaF/2 14.0 ± 2.7, NaF/3 12.6 ± 3.9, NaF/SnCl₂ 14.7 ± 5.1, AmF/SnF₂ 13.5 ± 4.8, AmF/NaF/SnCl₂ 12.4 ± 4.2, AmF/NaF/SnCl₂ + chitosan 6.6 ± 3.5 (except NaF/1 all p ≤ 0.01 compared to placebo). AmF/NaF/SnCl₂/chitosan was more effective than all other toothpastes (p ≤ 0.01 each). Sn on the enamel surface ranged between 1.3 ± 0.3 and 2.8 ± 0.04 wt.% with no obvious relationship with efficacy.

Conclusions

The NaF and Sn²⁺ toothpastes without chitosan exhibited similar anti-erosion and abrasion-prevention effects. The experimental Sn²⁺ formulation with chitosan revealed promising results similar to those of the positive control.

Clinical significance

NaF toothpastes offer a degree of protection against erosion/abrasion, which is likely sufficient for most subjects with average acid exposures. For patients with initial erosive lesions, however, more effective toothpaste is desirable. The combination of Sn²⁺ and a biopolymer appears promising in this context.