柠檬精油等天然产物对牙齿矿化作用的体外研究

发布时间：2018-07-03 02:50

本文选题：天然产物 + 早期龋　；参考：《天津医科大学》2014年硕士论文

【摘要】：目的龋病是在以细菌为主的多种因素影响下,牙体硬组织发生慢性进行性破坏的一种疾病。牙体硬组织始终处于脱矿和再矿化交替进行的化学动力过程中,若整个过程再矿化占主导地位,脱矿区域则被修复,反之则形成龋洞。因此,再矿化是对抗致龋因素攻击的一个很重要的自然修复过程。2004年开始,本课题组在国内外率先开展柠檬提取物防治龋病的系列研究,发现其对常见口腔致龋菌有抑制作用,具有抑制致龋菌早期粘附、产酸、产胞外多糖等致龋毒力因子的作用。研究发现柠檬精油(Lemon essential oil,LEO)主要成分是柠檬烯(Limonene,LIM)。茶多酚(Tea polyphenol,TP)是目前具有代表性的防龋天然产物,洗必泰(Chlorhexidine,CHX)是口腔常用的辅助抗菌药物。本课题在前期研究工作的基础上,从牙齿脱矿和再矿化角度评价LEO、LIM、TP和CHX在抑制脱矿和促进再矿化过程中的作用,并评价其可能的防龋效能,同时评价与氟化钠(NaF)的协同作用,为天然产物防龋的研发提供理论依据。方法 1.人工龋样本制备制备牛切牙牙釉质、牙本质样本,置于脱矿液中96小时形成人工龋标本。 2.实验分组空白对照组(negtive control,NC):去离子水阳性对照组：1mg/ml NaF；实验组：4.5m/ml LEO；4.5mg/ml LEO+1mg/ml NaF;21mg/ml LIM；21mg/ml LIM+1mg/ml NaF;2mg/ml TP;2mg/ml TP+1mg/ml NaF；2mg/ml CHX 3.pH循环实验 37℃条件下,将样本分别置于脱矿液中处理14h,实验溶液中处理2h,再矿化液中处理8h。每次处理前样本需要在缓冲液中震荡冲洗5min。整个pH循环持续8天。收集全部脱矿液及再矿化液,-20℃保存备用。 4.观察指标： (1)全自动生化分析仪测出脱矿液中钙、磷离子浓度,计算出钙、磷流出值,评价抑制脱矿作用。 (2)表面显微硬度仪测定样本脱矿前后和pH循环后的硬度值,计算出表面显微硬度恢复值,观察表面显微硬度的变化,评价再矿化作用。 (3)X射线能谱分析仪对样本表面钙磷比进行定性定量分析,评价再矿化作用。 (4)羟脯氨酸试剂盒检测牙本质再矿化液中羟脯氨酸浓度,羟脯氨酸含量反映牙本质胶原基质的保存量,以评价药物对胶原降解的影响。结果 1.脱矿液中钙、磷离子流出值 (1)牙釉质：LEO、LIM、TP和CHX钙、磷离子流出值与空白对照组比较,差异没有统计学意义(P0.05)。 (2)牙本质：LEO组钙离子流出值低于空白对照组,差异有统计学意义(P0.05)；LEO组磷离子流出值和LIM、TP及CHX组钙、磷离子流出值与空白对照组比较,差异没有统计学意义(P0.05)。 2.表面显微硬度值 LEO、LIM、TP及CHX组牙釉质及牙本质标本表面显微硬度恢复值与空白对照组比较差异没有统计学意义(P0.05)。 3.样本表面钙磷比 (1)牙釉质：LIM、TP和CHX组高于空白对照组,差异有统计学意义(P0.05)；LEO组与空白对照组比较,差异没有统计学意义(P0.05)。 (2)牙本质：LEO、LIM、TP和CHX组高于空白对照组且差异有统计学意义(P0.05)。 4.再矿化液中羟脯氨酸浓度LEO、LIM和CHX组低于空白对照组,差异有统计学意义(P0.05)；TP组与空白对照组比较,差异没有统计学意义(P0.05)。 5.与NaF联合应用LEO、LIM、TP和CHX与NaF联合应用没有发现有协同作用,与阳性对照组比较差异没有统计学意义(P0.05)。结论 1.LIM、TP和CHX具有稳定晶体结构,促进牛牙釉质再矿化作用； 2.LEO具有抑制牛牙本质脱矿的作用； 3.LEO、LIM、TP和CHX具有稳定晶体结构,促进牛牙本质再矿化作用； 4.LEO、LIM和CHX具有抑制牛牙本质胶原基质降解作用； 5.LEO、LIM、TP和CHX没有发现抑制牛牙釉质脱矿作用； 6.LEO、LIM和TP与NaF联合应用,没有发现与NaF有协同作用且不影响NaF作用效果； 7.与表面显微硬度检测方法相比,X线能谱分析是一项比较敏感的检测矿化作用指标。
[Abstract]:objective
Caries are a disease in which the hard tissue of the tooth has chronic progressive destruction under the influence of a variety of bacteria based factors. The hard tissue of the tooth body is always in the chemical dynamic process of alternation of demineralization and remineralization. If the remineralization is dominant in the whole process, the demineralization area is repaired, and on the contrary, the caries cave is formed. Therefore, remineralization is the remineralization. A very important natural restoration process against the attack of cariogenic factors began in.2004 years. The team took the lead in a series of studies on the prevention and control of caries by lemon extract at home and abroad, and found that it has a inhibitory effect on common dental cariogenic bacteria, and has the effect of inhibiting the early adhesion of cariogenic bacteria, acid producing, and extracellular polysaccharide, and so on. Lemon essential oil (LEO) is found to be the main component of limonene (Limonene, LIM). Tea polyphenols (Tea polyphenol, TP) are the representative natural products of anti caries, and chlorhexidine (Chlorhexidine, CHX) is a common oral antibacterials. This lesson is based on the prophase research work, from the angle of demineralization and remineralization of the teeth. The role of LEO, LIM, TP and CHX in inhibition of demineralization and remineralization was evaluated, and its possible caries prevention effectiveness was evaluated. At the same time, the synergistic effect of sodium fluoride (NaF) was evaluated to provide a theoretical basis for the development of natural products.
Method
1. artificial caries samples were prepared to prepare bovine enamel and dentin samples, and artificial caries specimens were formed in demineralized liquid for 96 hours.
2. the blank control group (negtive control, NC): the positive control group of deionized water: 1mg/ml NaF; the experimental group: 4.5m/ml LEO; 4.5mg/ml LEO+1mg/ml NaF; 21mg/ml LIM.
3.pH cycle experiment
At 37 C, the samples were treated with 14h in the demineralized solution, and the experimental solution was treated with 2H. The samples in the remineralized solution treated 8h. before each treatment needed to shake and flush the whole pH cycle for 8 days in the buffer solution. All the demineralized and remineralized fluids were collected and stored at -20 C.
4. the indicators of observation:
(1) automatic biochemical analyzer measured the concentration of calcium and phosphorus ions in the demineralization solution, calculated the outflow value of calcium and phosphorus, and evaluated the inhibition of demineralization.
(2) the surface microhardness tester was used to measure the hardness of the samples before and after the demineralization and the pH cycle. The recovery value of the surface microhardness was calculated, the change of the microhardness of the surface was observed and the remineralization was evaluated.
(3) X ray energy spectrum analyzer was used for qualitative and quantitative analysis of the ratio of calcium to phosphorus on the sample surface, and the remineralization was evaluated.
(4) hydroxyproline (hydroxyproline) kit was used to detect the concentration of hydroxyproline in dentin remineralized fluid. The content of hydroxyproline content reflected the preservation of dentin collagen matrix, so as to evaluate the effect of the drug on the degradation of collagen.
Result
1. the outflow value of calcium and phosphorus in the demineralized solution
(1) enamel: LEO, LIM, TP and CHX calcium and phosphorus outflow values were not significantly different from those in blank control group (P0.05).
(2) dentin: the outflow value of calcium ion in LEO group was lower than that in the blank control group, the difference was statistically significant (P0.05), and there was no statistically significant difference between the LEO group and the LIM, TP and CHX group, and the difference in the P + outflow value with the blank control group (P0.05).
2. surface microhardness value
There was no significant difference in the microhardness of enamel and dentin specimens between LEO, LIM, TP and CHX groups compared with blank control group (P0.05).
3. sample surface calcium and phosphorus ratio
(1) the tooth enamel: the LIM, TP and CHX groups were higher than the blank control group, the difference was statistically significant (P0.05), and there was no statistical difference between the LEO group and the blank control group (P0.05).
(2) dentin: LEO, LIM, TP and CHX were higher than those in the blank control group, and the difference was statistically significant (P0.05).
4. the concentration of hydroxyproline in the remineralized solution was LEO, and the group LIM and CHX were lower than the blank control group. The difference was statistically significant (P0.05), and there was no significant difference in the difference between the TP group and the blank control group (P0.05).
5. the combined application of LEO, LIM, TP and CHX combined with NaF did not show synergistic effect, and the difference was not statistically significant compared with the positive control group (P0.05).
conclusion
1.LIM, TP and CHX have stable crystal structure and promote remineralization of bovine enamel.
2.LEO has the effect of inhibiting the demineralization of bovine dentin.
3.LEO, LIM, TP and CHX have stable crystal structure and promote remineralization of bovine dentin.
4.LEO, LIM and CHX inhibited the degradation of bovine dentin collagen matrix.
5.LEO, LIM, TP and CHX did not inhibit demineralization of bovine enamel.
6.LEO, LIM and TP combined with NaF did not find synergistic effect with NaF and did not affect NaF effect.
7. compared with surface microhardness test, X-ray energy spectrum analysis is a sensitive indicator for detecting mineralization.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.1

【参考文献】