本文選題：天然產(chǎn)物 + 早期齲��； 參考：《天津醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的 齲病是在以細(xì)菌為主的多種因素影響下,牙體硬組織發(fā)生慢性進(jìn)行性破壞的一種疾病。牙體硬組織始終處于脫礦和再礦化交替進(jìn)行的化學(xué)動(dòng)力過(guò)程中,若整個(gè)過(guò)程再礦化占主導(dǎo)地位,脫礦區(qū)域則被修復(fù),反之則形成齲洞。因此,再礦化是對(duì)抗致齲因素攻擊的一個(gè)很重要的自然修復(fù)過(guò)程。2004年開(kāi)始,本課題組在國(guó)內(nèi)外率先開(kāi)展檸檬提取物防治齲病的系列研究,發(fā)現(xiàn)其對(duì)常見(jiàn)口腔致齲菌有抑制作用,具有抑制致齲菌早期粘附、產(chǎn)酸、產(chǎn)胞外多糖等致齲毒力因子的作用。研究發(fā)現(xiàn)檸檬精油(Lemon essential oil,LEO)主要成分是檸檬烯(Limonene,LIM)。茶多酚(Tea polyphenol,TP)是目前具有代表性的防齲天然產(chǎn)物,洗必泰(Chlorhexidine,CHX)是口腔常用的輔助抗菌藥物。本課題在前期研究工作的基礎(chǔ)上,從牙齒脫礦和再礦化角度評(píng)價(jià)LEO、LIM、TP和CHX在抑制脫礦和促進(jìn)再礦化過(guò)程中的作用,并評(píng)價(jià)其可能的防齲效能,同時(shí)評(píng)價(jià)與氟化鈉(NaF)的協(xié)同作用,為天然產(chǎn)物防齲的研發(fā)提供理論依據(jù)。 方法 1.人工齲樣本制備制備牛切牙牙釉質(zhì)、牙本質(zhì)樣本,置于脫礦液中96小時(shí)形成人工齲標(biāo)本。 2.實(shí)驗(yàn)分組空白對(duì)照組(negtive control,NC):去離子水陽(yáng)性對(duì)照組：1mg/ml NaF；實(shí)驗(yàn)組：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循環(huán)實(shí)驗(yàn) 37℃條件下,將樣本分別置于脫礦液中處理14h,實(shí)驗(yàn)溶液中處理2h,再礦化液中處理8h。每次處理前樣本需要在緩沖液中震蕩沖洗5min。整個(gè)pH循環(huán)持續(xù)8天。收集全部脫礦液及再礦化液,-20℃保存?zhèn)溆谩?4.觀察指標(biāo)： (1)全自動(dòng)生化分析儀測(cè)出脫礦液中鈣、磷離子濃度,計(jì)算出鈣、磷流出值,評(píng)價(jià)抑制脫礦作用。 (2)表面顯微硬度儀測(cè)定樣本脫礦前后和pH循環(huán)后的硬度值,計(jì)算出表面顯微硬度恢復(fù)值,觀察表面顯微硬度的變化,評(píng)價(jià)再礦化作用。 (3)X射線(xiàn)能譜分析儀對(duì)樣本表面鈣磷比進(jìn)行定性定量分析,評(píng)價(jià)再礦化作用。 (4)羥脯氨酸試劑盒檢測(cè)牙本質(zhì)再礦化液中羥脯氨酸濃度,羥脯氨酸含量反映牙本質(zhì)膠原基質(zhì)的保存量,以評(píng)價(jià)藥物對(duì)膠原降解的影響。 結(jié)果 1.脫礦液中鈣、磷離子流出值 (1)牙釉質(zhì)：LEO、LIM、TP和CHX鈣、磷離子流出值與空白對(duì)照組比較,差異沒(méi)有統(tǒng)計(jì)學(xué)意義(P0.05)。 (2)牙本質(zhì)：LEO組鈣離子流出值低于空白對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；LEO組磷離子流出值和LIM、TP及CHX組鈣、磷離子流出值與空白對(duì)照組比較,差異沒(méi)有統(tǒng)計(jì)學(xué)意義(P0.05)。 2.表面顯微硬度值 LEO、LIM、TP及CHX組牙釉質(zhì)及牙本質(zhì)標(biāo)本表面顯微硬度恢復(fù)值與空白對(duì)照組比較差異沒(méi)有統(tǒng)計(jì)學(xué)意義(P0.05)。 3.樣本表面鈣磷比 (1)牙釉質(zhì)：LIM、TP和CHX組高于空白對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；LEO組與空白對(duì)照組比較,差異沒(méi)有統(tǒng)計(jì)學(xué)意義(P0.05)。 (2)牙本質(zhì)：LEO、LIM、TP和CHX組高于空白對(duì)照組且差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 4.再礦化液中羥脯氨酸濃度LEO、LIM和CHX組低于空白對(duì)照組,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；TP組與空白對(duì)照組比較,差異沒(méi)有統(tǒng)計(jì)學(xué)意義(P0.05)。 5.與NaF聯(lián)合應(yīng)用LEO、LIM、TP和CHX與NaF聯(lián)合應(yīng)用沒(méi)有發(fā)現(xiàn)有協(xié)同作用,與陽(yáng)性對(duì)照組比較差異沒(méi)有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論 1.LIM、TP和CHX具有穩(wěn)定晶體結(jié)構(gòu),促進(jìn)牛牙釉質(zhì)再礦化作用； 2.LEO具有抑制牛牙本質(zhì)脫礦的作用； 3.LEO、LIM、TP和CHX具有穩(wěn)定晶體結(jié)構(gòu),促進(jìn)牛牙本質(zhì)再礦化作用； 4.LEO、LIM和CHX具有抑制牛牙本質(zhì)膠原基質(zhì)降解作用； 5.LEO、LIM、TP和CHX沒(méi)有發(fā)現(xiàn)抑制牛牙釉質(zhì)脫礦作用； 6.LEO、LIM和TP與NaF聯(lián)合應(yīng)用,沒(méi)有發(fā)現(xiàn)與NaF有協(xié)同作用且不影響NaF作用效果； 7.與表面顯微硬度檢測(cè)方法相比,X線(xiàn)能譜分析是一項(xiàng)比較敏感的檢測(cè)礦化作用指標(biāo)。
[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.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R781.1

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