氟斑牙祛色剂对牙体的安全性研究
摘要
目的：建立小鼠氟斑牙动物模型,用于研究氟斑牙祛色剂对牙体的安全性,并对氟斑牙祛色剂处理后的牙齿再矿化进行活体动态观察。
方法：(1)将3周龄的昆明小鼠65只随机分为2个实验组(每组30只)和1个对照组(5只),2个实验组小鼠分别给予浓度为100mg/L(高氟组)和50mg/L(低氟组)的氟化钠溶液自由饮用。对照组小鼠给予正常饮食,42天后,观察各组小鼠的上下切牙釉质透明度、光泽、形态,并在扫描电镜(SEM)下观察釉质表面结构。(2)将高氟组的小鼠随机分为5个实验组和1个对照组,每组5只,实验组分别用Beyond祛色剂Ⅰ液(A组),Beyond美白胶和冷光光照后再用Beyond祛色剂Ⅰ液(B组),Opalustre微研磨剂(C组),抛光杯单纯抛光(D组)处理切牙牙面后即刻处死及Beyond祛色剂Ⅰ液处理牙面后2周处死(E组),对照组不做任何处理处死,取每只小鼠的四颗切牙,分别制成相应的标本,进行扫描电镜观察,能谱仪分析,显微硬度测定,和牙体组织切片观察。(3)将低氟组的小鼠随机分为5个实验组和1个对照组,实验组分别用Beyond祛色剂Ⅰ液和Ⅱ液(A组),Beyond美白胶和冷光光照后再用Beyond祛色剂Ⅰ液和Ⅱ液(B组),Opalustre微研磨剂和再矿化剂(C组)处理切牙牙面后即刻处死及Beyond祛色剂Ⅰ液和Ⅱ液处理牙面后分别1周处死(D组)和2周处死(E组),对照组不做任何处理处死,取每只小鼠的四颗切牙,分别制成相应的标本,进行扫描电镜观察,能谱仪分析,显微硬度测定。所得数据采用SPSS10.0统计软件进行统计学分析。
结果：(1)42d后,观察小鼠上下切牙。高氟组小鼠切牙呈白垩色,釉质表面有大面积釉质缺损,严重者下切牙切端磨损并露髓,SEM观察结果显示釉质表面凹凸不平,在釉质缺损区可见条索状结构不规则排列；低氟组小鼠切牙釉质表面出现透明条带,或出现白垩色斑块,SEM观察结果显示釉质表面平滑,无缺损,蚀刻后可见釉柱大小和形状稍不规则,大部分按一定的方向排列；对照组小鼠切牙釉质呈半透明棕黄色,光滑而有光泽,SEM观察结果釉质表面光滑,结构致密,蚀刻后可见釉柱大小和形态几乎完全一致,并按一定的方向整齐排列。
(2)SEM观察显示A、B 2组釉质脱矿严重,呈粗糙老树皮状,C组亦有釉质脱矿,但脱矿程度较A、B 2组轻,E组牙釉质表面较光滑有矿物质均匀沉积；能谱分析结果示,A、B、C 3组钙、磷值及钙／磷比值均与D、E、对照组3组有显著性差异(P<0.05),A、B、C 3组间无差异(P>0.05),D、E、对照组3组间无差异(P>0.05)；显微硬度结果示,A、B、C 3组显微硬度均与D、E、对照组3组有显著性差异(P<0.05),A、B、C 3组间无差异(P>0.05),D、E、对照组3组间无差异(P>0.05)；切牙牙体组织学观察可见A、B、C 3组牙髓中有血管扩张充血现象,其中B组伴有组织水肿的现象,D、E、对照组3组牙髓组织未见异常。
(3)SEM观察结果示A、B、C、D、E各组牙表面均有矿物质沉积,A、C组矿物质沉积较B组多而均匀,D组较A组矿物质沉积更均匀,E组较D组矿物质沉积更多、更均匀、更光滑；能谱分析结果示,A、B、C、D、E、对照组6组钙、磷值及钙/磷比值无显著性差异(P>0.05)；显微硬度结果示,A、B、C、D、E、对照组6组显微硬度无显著性差异(P>0.05)。
结论：(1)小鼠饮用高浓度含氟水可建立典型的氟斑牙模型；
(2)氟斑牙祛色剂可以造成氟斑牙牙釉质面粗糙,矿物质含量及显微硬度下降；但经2周的唾液再矿化可使釉质硬度,钙、磷值及钙/磷比值达到脱色前水平；
(3)氟斑牙祛色剂对伴有釉质缺损的重度氟斑牙牙髓组织可造成充血反应,但为可逆性的；
(4)氟斑牙祛色剂配合再矿化剂治疗可快速恢复牙面的光滑度,硬度及钙、磷含量；唾液对牙齿的再矿化2周达到较理想水平。
Objective:To study the safety of fluoride-removing material on teeth and dynamically observe teeth remineralization in vivo after decoloring treatment on fluorosis mice tooth from fluorosis model.
Methods:(1) 65 3-week-old Kunming mice were randomly divided into 2 experimental groups (n=30) and a control group (n=5).The mice in two experimental groups was fed with distilled water containing 100mg/ L(high-fluoride group) and 50mg/L(low-fluoride group) of sodium fluoride respectively. The mice in control group was given normal diet.42 days later, all mice incisors change were observed about transparent, luster, shape and the enamel microstructure was observed by scanning electron microscopy (SEM). (2) The high-fluoride group was randomly and equally divided into 5 experimental groups and 1 control group. The mice incisors in 5 experimental groups were treated by different methods:A group treated by Beyond fluoride-removing material I solution, B group by Beyond fluoride-removing material I solution after beening used Beyond cold-light technique, C group by Opalustre micro abrasion, D group by polishing cup, all mice above immediately killed after treatment, E group by Beyond fluoride-removing material I solution and killed two weeks later. The control group was given no treatment. All mice were taken four incisors which were made into specimens respectively for electron microscopy scanning (SEM), energy dispersive X-ray spectroscopy (EDS), micro-hardness test and histological examination. (3) The low-fluoride group was also randomly and equally divided into 5 experimental groups and 1 control group. The mice incisors in 5 experimental groups were treated by different methods:A group treated by Beyond fluoride-removing materialⅠandⅡsolution, B group by Beyond fluoride-removing materialⅠandⅡsolution after beening used Beyond cold-light technique, C group by Opalustre micro abrasion and remineralizing solution, all mice above immediately killed after treatment, D and E group were treated as A group and respectively killed 1 week and 2 weeks later. The control group was given no treatment. All mice were taken four incisors which were made into specimens respectively for electron microscopy scanning (SEM), energy dispersive X-ray spectroscopy (EDS), micro-hardness test. All data were analyzed by software spss 10.0.
Results:(1) 42 days later, all chosen incisors of mice were examined. In high-fluoride group, the incisors enamel experienced changes in different degrees, such as scattered white spots, surface defects and even cut worn and pulp exposed; SEM observation showed the uneven enamel surface, cord-like structure arranged in irregular in enamel defects. In low-fluoride group, the incisors enamel surface showed transparent band or chalky patches; the SEM observation showed that the enamel surface was smooth and no defect. After enamel etching, some enamel rods slightly disordered and slightly irregular in size and shape, but most arranged as a certain direction. In control group, all incisors surface showed translucent enamel brown, smooth and shiny, the SEM observation showed that the enamel surface was smooth and dense, enamel rods were arranged in neat rows, size and shape was almost exactly the same and arranged according to a certain direction after enamel etching. (2) SEM observation showed that the enamel of A、B groups was severely demineralized and rough; C group was lightly demineralized than A、B group; E groups was mineral deposition uniformity and the enamel surface was smooth. There was significantly difference on the data about Calcium, phosphorus and calcium/phosphorus ratio among A、B、C、D、E、control group(P<0.05); whereas, there were no difference among A、B、C groups and no difference among D、E、control group (P >0.05). There was significantly difference on micro-hardness among A、B、C、D、E、control group(P<0.05); whereas, there was no difference among A、B、C groups and no difference among D、E、control group (P>0.05) A、B、C groups showed that pulp hypermia, further more B group was associated with tissue edema; whereas, D、E、control group showed normal pulp tissue. (3) SEM observation showed that there was mineral deposits on teeth surface in all 5 groups, which was more and even in A、C group than in the B group, more uniform mineral deposits in D group than in A group, more mineral deposits、more uniform and more smooth in E group than in D group. There was no significantly difference on the data about Calcium, phosphorus and calcium/phosphorus ratio among A、B、C、D、E、control group(P>0.05). And there was no significantly difference on micro-hardness among A、B、C、D、E、control group(P>0.05).
Conclusions:(1) The model of a typical dental fluorosis can be established by making mice drinking high concentrations of fluoride water.
(2) Fluorosis-removing material can cause enamel surface roughness and mineral content reduce and micro-hardness decreased, but the micro-hardness, calcium, phosphorus and calcium/phosphorus ratio can recover to the level before bleaching after 2 weeks.
(3) Fluorosis-removing material can cause congestive in the pulp tissue of severe dental fluorosis teeth, but the change is reversible.
(4) Fluorosis-removing material associated with remineralizing solution can rapidly recover tooth surface smoothness, micro-hardness and the content of calcium and phosphorus. In 2 weeks, teeth can be mineralized to a satisfactory level by saliva.
方法：(1)将3周龄的昆明小鼠65只随机分为2个实验组(每组30只)和1个对照组(5只),2个实验组小鼠分别给予浓度为100mg/L(高氟组)和50mg/L(低氟组)的氟化钠溶液自由饮用。对照组小鼠给予正常饮食,42天后,观察各组小鼠的上下切牙釉质透明度、光泽、形态,并在扫描电镜(SEM)下观察釉质表面结构。(2)将高氟组的小鼠随机分为5个实验组和1个对照组,每组5只,实验组分别用Beyond祛色剂Ⅰ液(A组),Beyond美白胶和冷光光照后再用Beyond祛色剂Ⅰ液(B组),Opalustre微研磨剂(C组),抛光杯单纯抛光(D组)处理切牙牙面后即刻处死及Beyond祛色剂Ⅰ液处理牙面后2周处死(E组),对照组不做任何处理处死,取每只小鼠的四颗切牙,分别制成相应的标本,进行扫描电镜观察,能谱仪分析,显微硬度测定,和牙体组织切片观察。(3)将低氟组的小鼠随机分为5个实验组和1个对照组,实验组分别用Beyond祛色剂Ⅰ液和Ⅱ液(A组),Beyond美白胶和冷光光照后再用Beyond祛色剂Ⅰ液和Ⅱ液(B组),Opalustre微研磨剂和再矿化剂(C组)处理切牙牙面后即刻处死及Beyond祛色剂Ⅰ液和Ⅱ液处理牙面后分别1周处死(D组)和2周处死(E组),对照组不做任何处理处死,取每只小鼠的四颗切牙,分别制成相应的标本,进行扫描电镜观察,能谱仪分析,显微硬度测定。所得数据采用SPSS10.0统计软件进行统计学分析。
结果：(1)42d后,观察小鼠上下切牙。高氟组小鼠切牙呈白垩色,釉质表面有大面积釉质缺损,严重者下切牙切端磨损并露髓,SEM观察结果显示釉质表面凹凸不平,在釉质缺损区可见条索状结构不规则排列；低氟组小鼠切牙釉质表面出现透明条带,或出现白垩色斑块,SEM观察结果显示釉质表面平滑,无缺损,蚀刻后可见釉柱大小和形状稍不规则,大部分按一定的方向排列；对照组小鼠切牙釉质呈半透明棕黄色,光滑而有光泽,SEM观察结果釉质表面光滑,结构致密,蚀刻后可见釉柱大小和形态几乎完全一致,并按一定的方向整齐排列。
(2)SEM观察显示A、B 2组釉质脱矿严重,呈粗糙老树皮状,C组亦有釉质脱矿,但脱矿程度较A、B 2组轻,E组牙釉质表面较光滑有矿物质均匀沉积；能谱分析结果示,A、B、C 3组钙、磷值及钙／磷比值均与D、E、对照组3组有显著性差异(P<0.05),A、B、C 3组间无差异(P>0.05),D、E、对照组3组间无差异(P>0.05)；显微硬度结果示,A、B、C 3组显微硬度均与D、E、对照组3组有显著性差异(P<0.05),A、B、C 3组间无差异(P>0.05),D、E、对照组3组间无差异(P>0.05)；切牙牙体组织学观察可见A、B、C 3组牙髓中有血管扩张充血现象,其中B组伴有组织水肿的现象,D、E、对照组3组牙髓组织未见异常。
(3)SEM观察结果示A、B、C、D、E各组牙表面均有矿物质沉积,A、C组矿物质沉积较B组多而均匀,D组较A组矿物质沉积更均匀,E组较D组矿物质沉积更多、更均匀、更光滑；能谱分析结果示,A、B、C、D、E、对照组6组钙、磷值及钙/磷比值无显著性差异(P>0.05)；显微硬度结果示,A、B、C、D、E、对照组6组显微硬度无显著性差异(P>0.05)。
结论：(1)小鼠饮用高浓度含氟水可建立典型的氟斑牙模型；
(2)氟斑牙祛色剂可以造成氟斑牙牙釉质面粗糙,矿物质含量及显微硬度下降；但经2周的唾液再矿化可使釉质硬度,钙、磷值及钙/磷比值达到脱色前水平；
(3)氟斑牙祛色剂对伴有釉质缺损的重度氟斑牙牙髓组织可造成充血反应,但为可逆性的；
(4)氟斑牙祛色剂配合再矿化剂治疗可快速恢复牙面的光滑度,硬度及钙、磷含量；唾液对牙齿的再矿化2周达到较理想水平。
Objective:To study the safety of fluoride-removing material on teeth and dynamically observe teeth remineralization in vivo after decoloring treatment on fluorosis mice tooth from fluorosis model.
Methods:(1) 65 3-week-old Kunming mice were randomly divided into 2 experimental groups (n=30) and a control group (n=5).The mice in two experimental groups was fed with distilled water containing 100mg/ L(high-fluoride group) and 50mg/L(low-fluoride group) of sodium fluoride respectively. The mice in control group was given normal diet.42 days later, all mice incisors change were observed about transparent, luster, shape and the enamel microstructure was observed by scanning electron microscopy (SEM). (2) The high-fluoride group was randomly and equally divided into 5 experimental groups and 1 control group. The mice incisors in 5 experimental groups were treated by different methods:A group treated by Beyond fluoride-removing material I solution, B group by Beyond fluoride-removing material I solution after beening used Beyond cold-light technique, C group by Opalustre micro abrasion, D group by polishing cup, all mice above immediately killed after treatment, E group by Beyond fluoride-removing material I solution and killed two weeks later. The control group was given no treatment. All mice were taken four incisors which were made into specimens respectively for electron microscopy scanning (SEM), energy dispersive X-ray spectroscopy (EDS), micro-hardness test and histological examination. (3) The low-fluoride group was also randomly and equally divided into 5 experimental groups and 1 control group. The mice incisors in 5 experimental groups were treated by different methods:A group treated by Beyond fluoride-removing materialⅠandⅡsolution, B group by Beyond fluoride-removing materialⅠandⅡsolution after beening used Beyond cold-light technique, C group by Opalustre micro abrasion and remineralizing solution, all mice above immediately killed after treatment, D and E group were treated as A group and respectively killed 1 week and 2 weeks later. The control group was given no treatment. All mice were taken four incisors which were made into specimens respectively for electron microscopy scanning (SEM), energy dispersive X-ray spectroscopy (EDS), micro-hardness test. All data were analyzed by software spss 10.0.
Results:(1) 42 days later, all chosen incisors of mice were examined. In high-fluoride group, the incisors enamel experienced changes in different degrees, such as scattered white spots, surface defects and even cut worn and pulp exposed; SEM observation showed the uneven enamel surface, cord-like structure arranged in irregular in enamel defects. In low-fluoride group, the incisors enamel surface showed transparent band or chalky patches; the SEM observation showed that the enamel surface was smooth and no defect. After enamel etching, some enamel rods slightly disordered and slightly irregular in size and shape, but most arranged as a certain direction. In control group, all incisors surface showed translucent enamel brown, smooth and shiny, the SEM observation showed that the enamel surface was smooth and dense, enamel rods were arranged in neat rows, size and shape was almost exactly the same and arranged according to a certain direction after enamel etching. (2) SEM observation showed that the enamel of A、B groups was severely demineralized and rough; C group was lightly demineralized than A、B group; E groups was mineral deposition uniformity and the enamel surface was smooth. There was significantly difference on the data about Calcium, phosphorus and calcium/phosphorus ratio among A、B、C、D、E、control group(P<0.05); whereas, there were no difference among A、B、C groups and no difference among D、E、control group (P >0.05). There was significantly difference on micro-hardness among A、B、C、D、E、control group(P<0.05); whereas, there was no difference among A、B、C groups and no difference among D、E、control group (P>0.05) A、B、C groups showed that pulp hypermia, further more B group was associated with tissue edema; whereas, D、E、control group showed normal pulp tissue. (3) SEM observation showed that there was mineral deposits on teeth surface in all 5 groups, which was more and even in A、C group than in the B group, more uniform mineral deposits in D group than in A group, more mineral deposits、more uniform and more smooth in E group than in D group. There was no significantly difference on the data about Calcium, phosphorus and calcium/phosphorus ratio among A、B、C、D、E、control group(P>0.05). And there was no significantly difference on micro-hardness among A、B、C、D、E、control group(P>0.05).
Conclusions:(1) The model of a typical dental fluorosis can be established by making mice drinking high concentrations of fluoride water.
(2) Fluorosis-removing material can cause enamel surface roughness and mineral content reduce and micro-hardness decreased, but the micro-hardness, calcium, phosphorus and calcium/phosphorus ratio can recover to the level before bleaching after 2 weeks.
(3) Fluorosis-removing material can cause congestive in the pulp tissue of severe dental fluorosis teeth, but the change is reversible.
(4) Fluorosis-removing material associated with remineralizing solution can rapidly recover tooth surface smoothness, micro-hardness and the content of calcium and phosphorus. In 2 weeks, teeth can be mineralized to a satisfactory level by saliva.
引文
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26 Smidt A, Weller D,Roman I, Gedalia I.Effect of bleaching agents on micorhardness and surface morphology. Am J Dent 1998,11:83-85.
27 Lopes GC, Bonissoni L, Baratieri LN,Vieira LC,Monteriro S Jr. Effect of bleaching agents on the hardness and morphology of enamel. J Esthet Restor Dent,2002:14(1):24-30.
28 Joiner A, Thakker G, Cooper Y. Evaluation of a 6% hisydrogen peroxide tooth whitening gel on enamel and dentine microhardness in vitro. Journal of Dentistry,2004,32 Suppll:27-34.
29 Leonard RH, Haywood VB, Phillips C. Risk factors for develping tooth sensivity and gingiral irritortion with nightguard vital bleaching [J]. Quint Int,1997,13(8):527-534.
30 Pohjola RM, Browning WD, Hackman ST, et al. Sensitivity and tooth whitening agents [J]. J Esthet Restor Dent,2002,14(2):85-91.
31 Browning WD, Chan DC, Frazier KB, et al. Comparison of the effectiveness and safety of carbamide peroxide whitening agents [J]. Quint Int,2004,35(9):693-698.
32赵奇,王君香,冯朝华.冷光牙齿漂白技术对活髓牙牙髓影响的病理观察[J].中华口腔医学杂志,2007,42(12)：718-719.
33 Hanks CT, Fat JC, Wataha JC. Cytotoxicity and dentin permeability of carbamide peroxide and hydrogen peroxide vital bleaching materials [J]. J Dent Res,1993,72(2):931-938.
34 Bowles WH, Thompson LR. Vital bleaching:the effects of heat and hydrogen peroxide on pulpal enzymes [J]. J Endodont,1986,12(3):108-112.
35 Boweles WH, Ugwunefi Z. Pulp chamber penetration by hydrogen peroxide fllowing vital bleaching procedures [J]. J Endodont.1987,13(8):375-377.
36 Hicks MJ, Silverstone LM. Acid-etching of carieslike lesions of enamel:a scanning electron microscopic study [J]. Caries Res,1984 (4),18:327.
37 Hattab FN, Wei SHY. Chemical changes and surface morphology of acid-etching of human enamel treated with topical fluoride agents in vitro [J]. Caries Res,1987,21(6):482-493.
38 Featherstone JDB, Rodgers BE, Smith MW. Physiochemical requirements for rapid remineralization of early carious lesions [J]. Caries Res,1981,15(6):221-225.
39 Attin T, Kielbassa AM, Schwanenberg M. Effect of fluoride treatment on remineralization of bleached enamel [J]. J Oral Rehabil,1997,24(4):282-286.
40朱强,王满恩,王勤,等.氟斑牙脱色治疗及再矿化修复的探讨[J].中华口腔医学杂志,1991,26(4)：195-198.
41樊明文主编.牙体牙髓病学.2003年,第2版,29-30.
42 Collys K,Cleymael R, Coomans D, et al. Rehardening of surface softened and surface etched enamel in vitro and by intraoral exposure [J]. Caries Res,1993,27(2):15-20.
1孙殿军.中国地方性氟中毒的病情及防治对策[J].中国地方病学杂志,2001,20(1)：1-3
2 Dayan D.Tooth discoloration extrinsic and intrinsic factors [J]. Quintessence Int,1988, 14(2):195-199.
3孙殿军,沈雁峰,赵新华,等.中国大陆地方性氟中毒病情动态及现状分析[J].中国地方病学杂志,2002,20(16)：429.
4 Derbesten PK, Grenshaw MA. The effect of chronic high fluoride levels on forming enamel in the rat. Archives of Oral Biology,1984,29(2):675-679.
5 Fejerskov O, Manji F, Baelum V. The nature and mechanisims of dental fluorosis in man [J]. J Dent Res,1990,69(Spec, Iss):692-700.
6 Fejerskov O, Yanagisawa T, Tohda 1, et al. Posteruptive changes in human dental fluorosis a histological and ultra-structural study [J]. Proc Finn Dent Sco,1991,87(5):607-619.
7任吉芳,吉媛媛.氟铁元素在氟斑牙着色釉质不同层次分析[J].世界元素医学杂志,1999,6(2)：16-19.
8任吉芳,吉媛媛.31种元素在着色氟牙症釉质中的定量分析[J].中华口腔医学杂志,1999,34(4)：229-231.
9任吉芳.山西省高氟区6-8岁人群着色型氟斑牙分布特点调查[J].中国地方病学杂志,1998,17(3)：397-400.
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12 Sucking G, Coote GE, Cutress TW, et al. Proton microprobe assessment of the distribution of fluoride in the enamel and dentine of developing central incisors of sheep and changes induced by daily fluoride supplements[J]. Arch Oral Biol,1995,40 (5):439-446.
13章魁华,于世凤.实验口腔病理学[M].北京：人民卫生出版社,2002：31-32.
14邓转云,黄瑞哲,郭永利.诱导大鼠下切牙氟斑牙模型所需饮用水氟浓度及时间的筛选[J].实验动物科学,2008,25(4)：12-15.
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18井凤桐,王强,刘天麟,等.过量氟对大鼠切牙基质金属蛋白酶-20及其组织抑制剂表达的影响[J].华西口腔医学杂志,2006,24(3)：199-201.
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28樊明文主编.牙体牙髓病学.人民卫生出版社,2000：108-110.
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36 Zekonis R, Matis BA, Cochran MA, et al. Clinical evaluation of in-office and at-home bleaching treatments [J]. Operative Dentistry,2003,28(2):114-121.
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52 Gerlach RW, Barker ML, Sagel PA. Comparative efficacy and tolerability of two direct-to-consumer tooth whitening systems [J]. Am J Dent,2001,14(5):267-272.
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54 Pohjola RM, Browning WD, Hackman ST, et al. Sensitivity and tooth whitening agents [J]. J Esthet Restor Dent,2002,14(2):85-91.
55 Browning WD, Chan DC, Frazier KB, et al. Comparison of the effectiveness and safety of carbamide peroxide whitening agents [J]. Quint Int,2004,35(9):693-698.
56 Hanks CT, Fat JC, Wataha JC. Cytotoxicity and dentin permeability of carbamide peroxide and hydrogen peroxide vital bleaching materials [J]. J Dent Res,1993,72(2):931-938.
57 Leonard RH, Haywood VB, Phillips C. Risk factors for develping tooth sensivity and gingiral irritortion with nightguard vital bleaching [J]. Quint Int,1997,13(8):527-534.
58 Cavalli V, Arrais CA, Giannini M, et al. High-concentrated carbamide peroxide bleaching agents effects on enamel surface [J]. J Oral Rehabil,2004,31(2):155-159.
59 Sulieman M, Addy M, Rees JS. Development and evaluation of a method in vitro to study the effectiveness of tooth bleaching [J]. J Dent,2003,31(6):415-422.
60 White DJ, Kozak KM, Zoladz JR, et al. Effects of tooth-whitening gels on enamel and dentin ultrastructure-a confocal laser scanning microscopy pilot study [J]. Compend Contin Educ Dent,2000,21(29):29-34.
61 Featherstone JD, Barrett-Vespone NA, Fried D, Kantorowitz Z, Seka W. CO2 laser inhibitor of artificial caries-like lesion progression in dental enamel [J]. J Dent Res.1998,77(6):1397-1403.
62 Smidt A, Weller D, Roman I, Gedalia I. Effect of bleaching agents on micorhardness and surface morphology [J]. Am J Dent,1998,11(2):83-85.
63 Lopes GC, Bonissoni L, Baratieri LN, Vieira LC, Monteriro S Jr. Effect of bleaching agents on the hardness and morphology of enamel [J]. J Esthet Restor Dent,2002,14(1):24-30.
64 Joiner A, Thakker G, Cooper Y. Evaluation of a 6% hisydrogen peroxide tooth whitening gel on enamel and dentine microhardness in vitro [J]. Journal of Dentistry,2004,32 Suppll:27-34.
65 Homewood C. Bonding to previously bleached teeth [J]. Aust Orthod J,2001,17 (1):27-34.
66 Ben-Amar A, Liberman R,Gorfil C, Bernstein Y. Effect of mouthguard bleaching on enamel surface [J]. Am J Dent 1995,8(1):29-32.
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68 Kum KY, Lim KR, Lee CY, et al. Effects of removing residual Peroxide and other oxygen radicals on the shear bond strength and failure modes at resin-tooth interface after tooth bleaching [J]. Am J Dent,2004,17(4):267-270.
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70 Sung EC, Chan SM, Mito R, Caputo AA. Effect of carbamide peroxide bleaching on the shear bong strength of composite to dental bongding agent enhanced enamel [J]. J Prosthet Dent,1999, 82 (4):595-9.
71 Sterret J, price RB, Bankey T. Effects of home bleaching on the tissues of the oral cavity [J]. J Can Dent Assoc,1995,61(5):412-417, quiz 420.
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74 Dahl JE, Becher R. Acute toxicity of carbamide peroxide and a commercially available tooth-bleaching agent in rats [J], J Dent Res,1995,74(2):710-714.
75 Hicks MJ, Silverstone LM. Acid-etching of carieslike lesions of enamel:a scanning electron microscopic study [J]. Caries Res,1984,18(4):327.
76 Hattab FN, Wei SHY. Chemical changes and surface morphology of acid-etching of human enamel treated with topical fluoride agents in vitro [J]. Caries Res,1987,21(6):482-493.
77Featherstone JDB, Rodgers BE, Smith MW. Physiochemical requirements for rapid remineralization of early carious lesions [J]. Caries Res,1981,15(6):221-225.
78 Attin T, Kielbassa AM, Schwanenberg M. Effect of fluoride treatment on remineralization of bleached enamel [J]. J Oral Rehabil,1997,24(4):282-286.
79朱强,王满恩,王勤,等.氟斑牙脱色治疗及再矿化修复的探讨[J].中华口腔医学杂志,1991,26(4)：195-198.
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7 Derbesten PK, Grenshaw MA. The effect of chronic high fluoride levels on forming enamel in the rat. Archives of Oral Biology,1984,29(2):675-679.
8刘易汉.地方性氟中毒防治指南[M].北京：人民卫生出版社,1988.15-16
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11 Fasanaro T.Bleaching teeth:history, chemicals, and methods used for common tooth discoloration [J]. J Esth Dent,1992,4(3):71-78.
12魏丽,闰秀娟,杨亚丽.Beyond冷光美白仪漂白四环素牙的疗效观察.中华现代临床医学杂志,2005,3(12)：1188
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15徐宇红,张疆弢,刘建国,等.组合酸对氟斑牙釉质蚀刻的电镜观察[J].遵义医学院学报,2007,30(2)：118-120.
16钟滨,钟伟主编.口腔组织学图谱.2006年,第1版,4-5.
17朱保.外用漂白剂对牙釉质影响的电镜下观察[J].中国美容医学,2001,10(5)：267-426.
18何克新,李兰,吴煜.唾液替代品对脱矿牙面再矿化的研究[J].广西医科大学学报,1999,16(6)769-771.
19岳松龄主编.现代龋病学.2009年,第一版,192.
20 Cavalli V, Arrais CA, Giannini M, et al. High-concentrated carbamide peroxide bleaching agents effects on enamel surface [J]. J Oral Rehabil,2004,31(2):155-159.
21 Sulieman M, Addy M, Rees JS. Development and evaluation of a method in vitro to study the effectiveness of tooth bleaching [J]. J Dent,2003,31 (6):415-422.
22 Akal N, Over H, Olmez A, Bondur H. Effects of carbamide peroxide containing bleaching agents on the morphology and subsurface hardness of enamel [J]. Journal of Clinical Pediatric Dentistry 2001, 25(3):293-296.
23 Featherstone JD, Barrett-Vespone NA, Fried D, Kantorowitz Z, Seka W. CO2 laser inhibitor of artificial caries-like lesion progression in dental enamel [J]. J Dent Res.1998,77(6):1397-1403.
24 Meredith N, Sherriff M, Setchell DJ, et al. Measurement of the microhardness and Young’s modulus of human enamel and dentin using an indentation technique[J]. Arch Oral Biol.1996,41(6):539-545.
25 Basting RT, Rodrigues AL, Serra MP. The effect of seven carbamide peroxide bleaching agents on enamel microhardness over time. Journal of the American Dental Association 2003; 134:1335-42.
26 Smidt A, Weller D,Roman I, Gedalia I.Effect of bleaching agents on micorhardness and surface morphology. Am J Dent 1998,11:83-85.
27 Lopes GC, Bonissoni L, Baratieri LN,Vieira LC,Monteriro S Jr. Effect of bleaching agents on the hardness and morphology of enamel. J Esthet Restor Dent,2002:14(1):24-30.
28 Joiner A, Thakker G, Cooper Y. Evaluation of a 6% hisydrogen peroxide tooth whitening gel on enamel and dentine microhardness in vitro. Journal of Dentistry,2004,32 Suppll:27-34.
29 Leonard RH, Haywood VB, Phillips C. Risk factors for develping tooth sensivity and gingiral irritortion with nightguard vital bleaching [J]. Quint Int,1997,13(8):527-534.
30 Pohjola RM, Browning WD, Hackman ST, et al. Sensitivity and tooth whitening agents [J]. J Esthet Restor Dent,2002,14(2):85-91.
31 Browning WD, Chan DC, Frazier KB, et al. Comparison of the effectiveness and safety of carbamide peroxide whitening agents [J]. Quint Int,2004,35(9):693-698.
32赵奇,王君香,冯朝华.冷光牙齿漂白技术对活髓牙牙髓影响的病理观察[J].中华口腔医学杂志,2007,42(12)：718-719.
33 Hanks CT, Fat JC, Wataha JC. Cytotoxicity and dentin permeability of carbamide peroxide and hydrogen peroxide vital bleaching materials [J]. J Dent Res,1993,72(2):931-938.
34 Bowles WH, Thompson LR. Vital bleaching:the effects of heat and hydrogen peroxide on pulpal enzymes [J]. J Endodont,1986,12(3):108-112.
35 Boweles WH, Ugwunefi Z. Pulp chamber penetration by hydrogen peroxide fllowing vital bleaching procedures [J]. J Endodont.1987,13(8):375-377.
36 Hicks MJ, Silverstone LM. Acid-etching of carieslike lesions of enamel:a scanning electron microscopic study [J]. Caries Res,1984 (4),18:327.
37 Hattab FN, Wei SHY. Chemical changes and surface morphology of acid-etching of human enamel treated with topical fluoride agents in vitro [J]. Caries Res,1987,21(6):482-493.
38 Featherstone JDB, Rodgers BE, Smith MW. Physiochemical requirements for rapid remineralization of early carious lesions [J]. Caries Res,1981,15(6):221-225.
39 Attin T, Kielbassa AM, Schwanenberg M. Effect of fluoride treatment on remineralization of bleached enamel [J]. J Oral Rehabil,1997,24(4):282-286.
40朱强,王满恩,王勤,等.氟斑牙脱色治疗及再矿化修复的探讨[J].中华口腔医学杂志,1991,26(4)：195-198.
41樊明文主编.牙体牙髓病学.2003年,第2版,29-30.
42 Collys K,Cleymael R, Coomans D, et al. Rehardening of surface softened and surface etched enamel in vitro and by intraoral exposure [J]. Caries Res,1993,27(2):15-20.
1孙殿军.中国地方性氟中毒的病情及防治对策[J].中国地方病学杂志,2001,20(1)：1-3
2 Dayan D.Tooth discoloration extrinsic and intrinsic factors [J]. Quintessence Int,1988, 14(2):195-199.
3孙殿军,沈雁峰,赵新华,等.中国大陆地方性氟中毒病情动态及现状分析[J].中国地方病学杂志,2002,20(16)：429.
4 Derbesten PK, Grenshaw MA. The effect of chronic high fluoride levels on forming enamel in the rat. Archives of Oral Biology,1984,29(2):675-679.
5 Fejerskov O, Manji F, Baelum V. The nature and mechanisims of dental fluorosis in man [J]. J Dent Res,1990,69(Spec, Iss):692-700.
6 Fejerskov O, Yanagisawa T, Tohda 1, et al. Posteruptive changes in human dental fluorosis a histological and ultra-structural study [J]. Proc Finn Dent Sco,1991,87(5):607-619.
7任吉芳,吉媛媛.氟铁元素在氟斑牙着色釉质不同层次分析[J].世界元素医学杂志,1999,6(2)：16-19.
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