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研究生:顏煒慶
研究生(外文):Wei-Ching Yen
論文名稱:鈣磷生醫材料於牙本質小管內的離子滲透及再結晶機制之研究
論文名稱(外文):Mechanism of Ion Permeability and Recrystallization of Calcium Phosphate Bio-materials in Dentinal tubules
指導教授:林俊彬林俊彬引用關係
口試委員:姜昱至章浩宏朱瑾
口試日期:2014-05-31
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:臨床牙醫學研究所
學門:醫藥衛生學門
學類:牙醫學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:137
中文關鍵詞:鈣磷生醫材料牙本質敏感症牙本質小管選擇性及通透性磷酸根及鈣離子動物實驗
外文關鍵詞:calcium phosphate biomaterialdentin hypersensitivitydentin tubules selectivity and permeabilitycalcium and phosphate ionanimal study
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目前市面上有許多號稱能有效緩解牙齒敏感症狀的商品,但卻沒有任何產品能立即而持久的療效。本團隊之前的研究顯示以明膠為模板的含碳酸鈣中孔洞二氧化矽複合材料 (簡稱中孔洞鈣矽材料,GCMS) 與 30%磷酸調拌,可以在 10 分鐘內,有效地阻塞牙本質小管達 30 μm,並且不會對牙齒造成酸蝕破壞, 而在細胞毒性實驗證實,中孔洞鈣矽材料的材料萃取液及中孔洞鈣矽材料/磷酸製劑經過牙本質屏障有好的細胞活性。因此,推測此材料具有應用於治療牙本質敏感症極大的潛力。
然而,對於此類鈣磷生醫材料於牙本質小管內的沉澱機制仍然是未知的。本團隊之前藉由Transwell dentin disc model 的結果發現,離子於酸性環境下較易通過0.2mm 的牙本質屏障且當離子整個帶電價數較高或是分子量較小時,其通過0.2 mm牙本質屏障的速度也較快,不過,針對離子於牙本質小管的通透性,再結晶物內鈣離子的來源以及如何控制材料於牙本質小管內的再結晶長度則仍然未知。做為一個理想的牙科臨床治療材料,本團隊希望進一步了解其在牙本質小管內沉澱結晶的機制,並了解牙本質小管對於離子的通透性以及其在活體內的效用性,以使未來能更容易將治療藥物帶入牙本質小管及牙髓神經。
因此,本研究的目的是:
1.了解含碳酸鈣中孔洞二氧化矽複合材料牙本質小管內的離子的通透性及
2.找出時間因子對於材料於牙本質小管內再結晶的影響
3.找出再結晶物內鈣離子的來源
4.利用動物實驗證實本團隊的GCMS 材料於活體內的效用
第一部分以Transwell dentin disc model 及離子層析儀確認GCMS/30%磷酸,中孔洞鍶氧化矽/30%磷酸,30%磷酸以及氫氧化鈣/去離子水製劑經由 0.25 mm 牙齒滲透所釋放的陰陽離子濃度。接著,再用 SEM 觀察這些牙本質試片。
第二部分以GCMS/30%磷酸製劑塗抹於牙本質試片上,並於四個不同的時間點將牙齒劈開並於SEM下觀察其再結晶的情況。另外,再以EDS分析再結晶物的成份以及其鈣磷比值。
第三部份以中孔洞鍶氧化矽/30%磷酸製劑塗抹於牙本質試片上,並以EDS分析再結晶物的成份,以確定再結晶物內是否有鈣離子的存在。
第四部分則進行動物實驗以評估GCMS/30%磷酸,中孔洞鍶氧化矽/30%磷酸製劑在活體的效用性。
結論:(1)鈣離子本身不易穿透牙本質屏障,酸性環境的磷酸根離子可能扮演前導的角色,磷酸根離子濃度提高而影響電位,對於鈣離子產生引力,使鈣離子進入牙本質小管,而後由於鈣離子以及磷酸根離子濃度超過Ksp而形成沉澱結晶。
(2)材料放置的時間愈長,則產生之結晶愈長,因此推測可以藉由時間因子來控制再結晶的長度。
(3)再結晶中的鈣離子有部份來自於牙齒。
(4)GCMS/30%磷酸,中孔洞鍶氧化矽/30%磷酸製劑在活體內有牙髓腔靜水壓存在下,仍可在牙本質小管內產生超過 60 μm 的結晶,並在不破壞牙本質的情形下有效封閉牙本質小管。

There are various kinds of commercial products claimed to be able to relieve the symptoms of dentin hypersensitivity, but none of the products have immediate and long-lasting effects. Our previous study showed that gelatin-templated calcium mesoporous silicate (GCMS) with 30% H3PO4 could efficiently occlude dentinal tubules by precipitation thus have great potential in treating dentin hypersensitivity. Also, GCMS with 30% H3PO4 could show great biocompatibility through dentin disc barrier. To be an idea dental material, it is important to find out the mechanism of ion permeability and recrrstaliiizaton of calcium phosphate biomaterial in dentinal tubule.
Therefore, research goals of the present study is:
1. To find out the mechanism of ion permeability and recrystallization of CaCO3@ Mesoporous Silica-Phosphate in dentinal tubule
2. To find out the effect of time factor to the recrystallization of CaCO3@ Mesoporous Silica-Phosphate in dentinal tubule
3. To find out the source of calcium in the recrystallization in dentinal tubule
4. To evaluate the efficacy in vitro by animal study
This research comprised of four parts. First, we used transwell dentin disc model and ion chromatography to measure the ion permeability of GCMS / 30% H3PO4 , SrCO3 mesoporous silicates / 30% H3PO4 , 30 % H3PO4 and calcium hydroxide. The second part we applied GCMS mixed with 30% H3PO4 in the dentin disc and observed the recrystallization in the dentinal tubule by SEM at four different time points. Furthermore, EDS was used to analyze the component of recrystallization. At third part, we applied strontium mesoporous silicates mixed with 30% H3PO4 in the dentinal tubule to find out the source of calcium in the recrystallization by using EDS. Fourth, we used animal study to evaluate the efficacy of GCMS / 30% H3PO4 , SrCO3 mesoporous silicates / 30% H3PO4 in vivo.
In conclusion: (1)Calcium ion was not easily penetrate through the dentin barrier .PO43- could be a pilot to usher Ca2+ into dentinal tubules induce recrystallization.(2)The length of the recrystallization and the placement period of material are in direct proportion. (3) Partial calcium ion in the recrystallization was from the tooth. (4) GCMS/ 30% H3PO4 , SrCO3 mesoporous silicates / 30% H3PO4 had great efficacy in vivo.

中文摘要 ................................................................. I
ABSTRACT ..........................................................III
目錄 ........................................................................ V
圖次 ...................................................................... IX
表次 .................................................................... XIII
第一章 前言 ...........................................................1
第二章 文獻回顧 ...................................................3
2.1 牙本質過敏症的定義 ...................................................................................3
2.2 牙本質過敏症盛行率 .................................................................................3
2.3 牙本質過敏症致病因子 ..............................................................4
2.3.1 牙本質曝露 ....................................................................................................4
2.3.2 牙本質小管開口通暢 ....................................................................................5
2.4 牙本質過敏症的機制 ..............................................................................7
2.4.1 造牙本質細胞感覺接受器理論 ....................................................................7
2.4.2 神經支配理論 ................................................................................................7
2.4.3 流體力學理論 ................................................................................................8
2.5 牙本質滲透性 ................................................................................................8
2.6 牙本質過敏症治療材料與方法 .....................................................................10
2.6.1 化學性減敏方法 ..........................................................................................11
2.6.2 物理性減敏方法 ..........................................................................................15
2.7 生醫玻璃的應用 ...........................................................................18
2.8 孔洞材料原理 ...........................................................................19
2.8.1 中孔洞分子篩 ............................................................................19
2.8.2 中孔洞分子篩的性質與應用 .............................................21
2.9 &;#63756;米孔洞材料 ............................................................................22
2.9.1 中孔洞二氧化矽模板 ..................................................................................22
2.9.2 結合&;#63754;屬氧化物與中孔洞二氧化矽 ..........................................................23
2.10 牙本質小管之離子通透性及選擇性 ................................................24
2.10.1 擴散傳送 ..................................................................................24
2.10.2 對流傳送 .........................................................................................25
2.10.3 牙齒硬組織的離子通透性 ........................................................26
2.11 牙本質小管帶電情形 .....................................................................26
2.12 離子交換層析原理 .....................................................................27
2.12.1 基礎原理 ......................................................................................27
2.12.2 離子交換劑選擇 ........................................................................................28
2.13 動物實驗 ...................................29
第三章 動機與目的 .......................................................................................31
第四章 材料與方法 ..............................................................................................32
4.1 製備去敏感材料 .................................................................................32
4.1.1 以明膠為模版之中孔洞鈣矽材料以及中孔洞鍶矽材料之製作 ..............32
4.1.2 製備取代30%磷酸製劑之抗敏感製劑 .....................................................33
4.1.3 製備實驗所需各種基本溶液 ......................................................................33
4.2 中孔洞鈣矽材料/磷酸製劑及中孔洞鍶矽材料/磷酸製劑之性質分析 ............33
4.2.1 測量中孔洞鈣矽材料粉末與不同比例磷酸(30%)混合製劑之酸鹼值 ....33
4.2.2 測量中孔洞鍶矽材料粉末與不同比例磷酸製劑之酸鹼值 ......................34
4.3 牙本質小管之離子通透性及選擇性 .............................................................34
4.3.1 Transwell dentin disc model 之製備 ...........................................................34
4.3.2 待測溶液之放置 ..........................................................................................35
4.3.3 鈣離子及鍶離子以及磷酸根離子濃度之測量及統計 ..............................35
4.3.4 Transwell dentin disc model 之牙本質試片SEM &; EDS 觀察 ................36
4.4 觀察及比較放置含中孔洞鈣矽材料/磷酸製劑之時間長短對牙本質小管封閉
效果 ...................................................................36
4.4.1 牙本質試片之製備 ......................................................................................36
4.4.2 評估放置中孔洞鈣矽材料/磷酸混合製劑之時間對牙本質試片的影響 .37
4.4.3 掃描式電子顯微鏡 (SEM) 進行樣本觀察 ...............................................37
4.4.4 能量散射光譜分析儀 (EDS) 觀察 .........................................................38
4.5 觀察及比較放置含中孔洞鍶矽材料/磷酸製劑對牙本質小管封閉效果以及結
晶物之分析 .......................................................38
4.5.1 掃描式電子顯微鏡之觀察評估含中孔洞鍶矽材料/磷酸製劑對牙本質小
管的封閉效果 ........................................................................................................38
4.5.2 能量散射光譜分析儀 (EDS) 觀察 .........................................................39
4.6 動物實驗 ....................................................39
4.6.1 實驗動物之選擇與照顧 ..............................................................................39
4.6.2 實驗動物之麻醉 ..........................................................................................39
4.6.3 實驗步驟 ......................................................................................................40
第五章 結果 .........................................................42
5.1 中孔洞鈣矽材料/磷酸製劑及中孔洞鍶矽材料/磷酸製劑之性質分析 ............42
5.1.1 測量中孔洞鈣矽材料粉末與不同比例磷酸(30%)混合製劑之酸鹼值 ....42
5.1.2 測量中孔洞鍶矽材料粉末與不同比例磷酸(30%)混合製劑之酸鹼值 ....42
5.2 牙本質小管之離子通透性及選擇性觀察 ...........................................................43
5.3 Transwell dentin disc model 支牙本質試片SEM 觀察以及在結晶物成份之分
析 .....................................................................44
5.3.1 中孔洞鈣矽材料/磷酸製劑之牙本質試片觀察 ..........................................44
5.3.2 中孔洞鍶矽材料/磷酸製劑之牙本質試片觀察 .........................................45
5.3.3 磷酸製劑之牙本質試片觀察 ......................................................................45
5.3.4 氫氧化鈣/水製劑之牙本質試片觀察 .........................................................45
5.3.5 牙本質小管內再結晶物之成份分析 ..........................................................46
5.4 評估放置中孔洞鈣矽材料/磷酸混合製劑之時間長短對牙本質試片的影響 ….47
5.4.1 掃描式電子顯微鏡 (SEM) 進行樣本觀察 ...............................................47
5.4.2 能量散射光譜分析儀 (EDS)觀察 .............................................................48
5.5 觀察及比較放置含中孔洞鍶矽材料/磷酸製劑對牙本質小管封閉效果以及結
晶物之分析 .......................................................50
5.5.1 掃描式電子顯微鏡觀察評估製劑對牙本質小管的封閉效果 ...................50
5.5.2 牙本質小管內再結晶物之成份分析 ..........................................................51
5.6 動物實驗: 活體有效性測試 ...................................................................52
5.6.1 掃描式電子顯微鏡觀察評估製劑對牙本質小管的封閉效果 ..................52
5.6.2 牙本質小管內再結晶物之成份分析 ...........................................................53
第六章 討論 .........................................................56
6.1 牙本質小管之離子通透性及選擇性觀察 ...............................................56
6.1.1 中孔洞鈣氧化矽材料/磷酸製劑材料塗抹於牙本質試片所析出的離子 .56
6.1.2 中孔洞鍶氧化矽材料/磷酸製劑材料塗抹於牙本質試片所析出的離子 ..56
6.1.3 30%磷酸製劑材料塗抹於牙本質試片所析出的離子 ............................57
6.1.4 氫氧化鈣材料以及控制組(二次水)塗抹於牙本質試片所析出的離子
......................................................................57
6.1.5 材料塗抹於牙本質試片的結晶沉澱機制 ................................................58
6.2 Transwell dentin disc model 之牙本質試片SEM 觀察以及再結晶物成份之分
析 .....................................................................59
6.2.1 Transwell dentin disc model 之牙本質試片SEM 觀察 ............................59
6.2.2 Transwell dentin disc model 之牙本質試片再結晶成份分析 ...................59
6.3 評估及比較放置中孔洞鈣矽材料/磷酸混合製劑之時間長短對牙本質試片的
影響 ...................................................................60
6.3.1 掃描式電子顯微鏡觀察評估製劑對牙本質小管的封閉效果 ...................60
6.3.2 增加放置材料的時間如何幫助結晶的生成 ............................................61
6.3.3 牙本質小管內再結晶物之成份分析 .......................................................61
6.4 觀察及比較放置含中孔洞鍶矽材料/磷酸製劑對牙本質小管封閉效果以及結
晶物之分析 .......................................................61
6.4.1 觀察及比較放置含中孔洞鍶矽材料/磷酸製劑對牙本質小管封閉效果 .61
6.4.2 觀察及比較放置含中孔洞鍶矽材料/磷酸製劑對牙本質小管再結晶物之
分析 ..............................................................62
6.5 動物實驗:活體效用性 .........................................................63
6.5.1 掃描式電子顯微鏡觀察評估製劑對牙本質小管的封閉效果 ...................63
6.5.2 觀察及比較放置製劑對牙本質小管再結晶物之分析 ..............................64
第七章 結論 ......................................................65
第八章 未來研究方向 ...............................................................67
參考文獻 ...............................................................69
附圖 .......................................................................86
附表 .........................................................................132


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