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研究生:溫如琳
研究生(外文):Ju-Lin Wen
論文名稱:碳酸鈣中孔洞鈣矽材料與磷酸混合製劑於牙本質小管內的通透性及再結晶機制
論文名稱(外文):Mechanism of Ion Permeability & Recrystallization of CaCO3@ Mesoporous Silica-Phosphate in Dentinal Tubules
指導教授:林俊彬林俊彬引用關係
指導教授(外文):Chun-Pin Lin
口試委員:姜昱至朱瑾
口試委員(外文):Yu-Chih ChiangJinn P. Chu
口試日期:2013-06-08
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:2013
畢業學年度:101
語文別:中文
論文頁數:159
中文關鍵詞:中孔洞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
外文關鍵詞:Mesoporousdentin hypersensitivitydentin tubules selectivity and permeabilitycalcium and phosphate ion
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牙本質敏感症的治療截至目前,仍沒有一個令人滿意的臨床結果,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,具有應用於臨床,治療牙本質敏感症極大的潛力。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,而中孔洞鈣矽材料/磷酸製劑的材料萃取液則有較高的細胞毒性。
本團隊之前離子層析儀及Transwell dentin disc model的結果,氫氧化鈣/去離子水製劑不會經由0.2 mm的牙本質屏障釋放出鈣離子,而中孔洞鈣矽材料/磷酸溶液則能經由0.2 mm的牙本質屏障釋放出鈣離子及磷酸根離子,可初步推測中孔洞鈣矽材料/磷酸製劑形成沉澱結晶的機制:酸性提高而使電勢能提高,對於鈣離子產生引力,使鈣離子進入牙本質小管,而後由於牙齒有緩衝的作用,pH值升高形成沉澱結晶。
但做為一個理想的牙科臨床治療材料,30%磷酸在臨床試驗的實行上有其困難,生物相容性更是不可忽視的問題,並且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及選擇性,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. 找出影響牙本質小管通透性的因素,及牙本質小管是否有離子選擇性?
2. 找出是否有其它材料能替代原本的30%磷酸配方,一樣能達到阻塞牙本質小管的效果,並期待有更好的生物相容性。
3. 證實本團隊的GCMS材料,在使用後能增加牙本質硬度。
第一部分,以New Transwell dentin disc model及離子層析儀確認0.5 M磷酸二氫鉀溶液,0.5 M氯化鈣溶液,及GCMS/30%磷酸製劑經由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和磷酸二氫鈉溶液,磷酸二氫鉀溶液,及調整過pH值的磷酸二氫鉀溶液以不同比例混合;或以本團隊和西德有機醫藥生技公司合作製成的GCMS paste及磷酸二氫鈉paste以不同比例混合,再分成不同天數組別,塗抹在牙本質試片上,用SEM觀察其成效。
第三部分,以GCMS和30%磷酸製劑以粉液比1:2混合,並塗抹在牙本質試片上三次,第三次以pH值9的鹼性水使其沈澱。之後再以硬度計測試牙本質的硬度是否有增加。
結論:(1) 牙本質小管的各離子通透性在酸性環境下較好。(2) 在中性與酸性環境下,正電荷離子在此TW DD model較負電荷離子易於通過牙本質小管。(3) 在此TW DD model,當離子整個帶電價數較高時,通過速度也較快。(4) 磷酸二氫鈉比起磷酸二氫鉀更適合取代之前的30%磷酸,與GCMS混合,以治療牙本質過敏症。(5) 除了適當的pH值,材料本身的鈣磷莫耳濃度比對於牙本質小管的再結晶作用亦是重要的。(6) GCMS與30﹪磷酸的混合製劑,塗抹在牙本質上,可增加牙本質的硬度,也可能提高抗刷耗的功能。

Dentin hypersensitivity is a common encountered chief complaint for dental general practitioner, but it is also a troublesome issue. 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 and other comparisans obtain similar results. The transwell dentin disc model and ion chromatography revealed that calcium hydroxide with deionized water would not release calcium ion through 0.2 mm dentin disc. In contrast, GCMS with 30% H3PO4 could obtain phosphoric acid and calcium.
Therefore, research goals of the present study is:
1. To find out the factors that affect ion permeability & selectivity in dentinal tubules.
2. To find a replacement for 30% H3PO4.
3. To prove our material can increase microhardness of dentin.
This research comprised of three parts. First, we assess the differences of ion permeability and selectivity through dentin disks between pH value, ionic charge, valence electron.
The second part investigated the obturating effects in dentinal tubules of NaH2PO4 and KH2PO4 saturated solution when they are replaced the original 30% H3PO4 and mixed with GCMS. Futherly, our group fabricated a paste that include GCMS and NaH2PO4, then we also evaluate the obturating effects in dentinal tubules of the mixture.
In the third part, we estimated the microhardness of the dentin surface before and after applying GCMS/30% H3PO4 mixture. If the application of materials can increase the hardness of dentin, it may also raise up the possibility of anti-abrasion.
In conclusion:(1) Ion permeability is better in acidic condition. (2) Positive charge passes dentinal tubules easier in acidic & neutral condition than negative charge at this model. (3) When there are more valence electrons, the ions may pass the dentinal tubules easier. (4) According to their obturated effects, NaH2PO4 is a better replacement for H3PO4 than KH2PO4. (5) Other than pH gradient, Ca2+/PO43- ratio is also important in recrystallization at dentinal tubules. (6) GCMS/H3PO4 mixture can increase microhardness of dentin.

中文摘要 i
Abstract iii
目錄 v
圖次 xii
表次 xvi
第一章 前言 1
第二章 文獻回顧 4
2.1 牙本質過敏症的定義 4
2.2 牙本質過敏症盛行率 4
2.3 牙本質過敏症致病因子 5
2.3.1 牙本質曝露 5
2.3.2 牙本質小管開口通暢 6
2.4 牙本質過敏症的機制 8
2.4.1 造牙本質細胞感覺接受器理論 (Odontoblasts as receptors theory) 8
2.4.2 神經支配理論 (Direct innervation of dentin theory) 8
2.4.3 流體力學理論 (Hydrodynamic theory) 9
2.5 牙本質滲透性 10
2.6 牙本質過敏症治療材料與方法 12
2.6.1 化學性減敏方法 12
2.6.2 物理性減敏方法 17
2.7 生醫玻璃的應用 20
2.8 孔洞材料原理 21
2.8.1 中孔洞分子篩 21
2.8.2 中孔洞分子篩的性質與應用 23
2.9 奈米孔洞材料 23
2.9.1 中孔洞二氧化矽模板 23
2.9.2 結合金屬氧化物與中孔洞二氧化矽 24
2.10 牙本質小管之離子通透性及選擇性 26
2.10.1 擴散傳送 (Diffusion transport) 26
2.10.2 對流傳送 (Convection transport) 26
2.10.3 牙齒硬組織的離子通透性 ( Ion permeability of dental hard tissues) 28
2.11 牙本質小管帶電情形 28
2.12 離子交換層析原理 (Ion exchange chromatography) 29
2.12.1 基本原理 29
2.12.2 離子交換劑的選擇 30
第三章 動機與目的 32
第四章 材料與方法 33
4.1 製備去敏感材料 33
4.1.1 以明膠為模版之中孔洞鈣矽材料之製作 - Ca:Si= 1:1 33
4.1.2 製備實驗所需各種基本溶液 34
4.2 牙本質小管之離子通透性及選擇性 35
4.2.1 Transwell dentin disc model之製備 35
4.2.2 待測溶液之放置 36
4.2.3 陽離子及陰離子濃度之測量及統計 37
4.2.4 Transwell dentin disc model之牙本質試片SEM觀察 37
4.3 觀察及比較不同含中孔洞鈣矽材料/磷酸鹽製劑對牙本質小管封閉效果 38
4.3.1 牙本質試片之製備 38
4.3.2 評估不同比例中孔洞鈣矽材料/磷酸二氫鈉飽和溶液 混合製劑對牙本質試片的影響 38
4.3.3 評估不同比例中孔洞鈣矽材料/磷酸二氫鉀飽和溶液 混合製劑對牙本質試片的影響 39
4.3.4 評估不同比例中孔洞鈣矽材料/調整過pH值之磷酸二氫鉀飽和溶液 混合製劑對牙本質試片的影響 39
4.3.5 評估不同比例中孔洞鈣矽材料paste /磷酸二氫鈉paste混合製劑對牙本質試片的影響 39
4.3.6 評估新配方中孔洞鈣矽材料paste /磷酸二氫鈉paste混合製劑於不同天數對牙本質試片的影響 40
4.3.7 評估Sensodyne Repair paste於不同天數對牙本質試片的影響 40
4.3.8 控制組之牙本質試片處理 40
4.3.9 掃描式電子顯微鏡 (SEM) 進行樣本觀察 40
4.4 中孔洞鈣矽材料/各種磷酸鹽製劑之性質分析 42
4.4.1 測量中孔洞鈣矽材料粉末與不同比例磷酸二氫鈉飽和溶液 混合製劑之酸鹼值 42
4.4.2 測量中孔洞鈣矽材料粉末與不同比例磷酸二氫鉀飽和溶液 混合製劑之酸鹼值 42
4.4.3 測量中孔洞鈣矽材料粉末與不同比例調整過pH值之磷酸二氫鉀飽和溶液 混合製劑之酸鹼值 42
4.4.4 測量中孔洞鈣矽材料粉末paste與不同比例磷酸二氫鈉paste混合製劑之酸鹼值 43
4.4.5 測量新配方中孔洞鈣矽材料粉末paste /磷酸二氫鈉paste 1:1混合製劑之酸鹼值 43
4.4.6 測量Sensodye Repair paste之酸鹼值 43
4.5 評估中孔洞鈣矽材料/30%磷酸 粉液比1:2混合製劑對牙本質硬度的影響 43
4.5.1 牙本質壓片model之製作 44
4.5.2 中孔洞鈣矽材料/30%磷酸製劑之塗抹 44
4.5.3 硬度測試 44
第五章 結果 45
5.1 陰陽離子穿透Transwell DD Model之釋放量 45
5.1.1 比較不同pH值0.5 M不同待測溶液之各離子穿透累積釋放量 45
5.1.2 比較同一待測溶液之不同離子累積釋放量 46
5.1.3 比較正電荷離子及負電荷離子之累積穿透釋放量 47
5.1.4 比較各pH值下的不同價數/不同正電荷離子之累積穿透釋放量 47
5.1.5 比較不同價數/不同及同一負電荷離子之累積穿透釋放量 48
5.1.6 碳酸鈣中孔洞鈣矽材料/30%磷酸粉液比1:2之混合製劑的離子穿透累積釋放量 49
5.2 Transwell DD model牙本質試片SEM觀察 50
5.2.1 pH值為4之磷酸二氫鉀溶液穿透牙本質試片觀察 50
5.2.2 pH值為7之磷酸二氫鉀溶液穿透牙本質試片觀察 50
5.2.3 pH值為10之磷酸二氫鉀溶液穿透牙本質試片觀察 50
5.2.4 pH值為4之氯化鈣溶液穿透牙本質試片觀察 50
5.2.5 pH值為7之氯化鈣溶液穿透牙本質試片觀察 50
5.2.6 pH值為10之氯化鈣溶液穿透牙本質試片觀察 51
5.2.7 中孔洞鈣矽材料/30﹪磷酸粉液比1:2混合製劑之穿透牙本質試片觀察 51
5.3 觀察及比較不同比例含中孔洞鈣矽材料/磷酸二氫鈉飽和溶液混合製劑的酸鹼值,並評估其對牙本質小管封閉效果 51
5.3.1 中孔洞鈣矽材料及不同比例磷酸二氫鈉飽和溶液 混合製劑之酸鹼值 51
5.3.2 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
5.4 觀察及比較不同比例含中孔洞鈣矽材料/磷酸二氫鉀飽和溶液混合製劑的酸鹼值,並評估其對牙本質小管封閉效果 53
5.4.1 中孔洞鈣矽材料及不同比例磷酸二氫鉀飽和溶液 混合製劑之酸鹼值 53
5.4.2 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 53
5.5 觀察及比較不同比例含中孔洞鈣矽材料/調整過pH值之磷酸二氫鉀飽和溶液 混合製劑的酸鹼值,並評估其對牙本質小管封閉效果 54
5.5.1 中孔洞鈣矽材料及不同比例調整過pH值之磷酸二氫鉀飽和溶液 混合製劑之酸鹼值 54
5.5.2 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 55
5.6 觀察及比較不同比例含中孔洞鈣矽材料paste /磷酸二氫鈉paste混合製劑的酸鹼值,並評估其對牙本質小管封閉效果 56
5.6.1 觀察及比較不同比例含中孔洞鈣矽材料paste /磷酸二氫鈉paste混合製劑之酸鹼值 56
5.6.2 掃描式電子顯微鏡之觀察評估製劑對牙本質小管的封閉效果 57
5.7 觀察及比較新配方含中孔洞鈣矽材料paste /磷酸二氫鈉paste 1:1混合製劑的酸鹼值,並評估其對牙本質小管於不同天數封閉效果 57
5.7.1 觀察新配方含中孔洞鈣矽材料paste磷酸二氫鈉paste 1:1 混合製劑之酸鹼值 58
5.7.2 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 58
5.8 觀察及比較Sensodyne Repair paste的酸鹼值,並評估其對牙本質小管於不同天數封閉效果 59
5.8.1 觀察Sensodyne Repair paste之酸鹼值 59
5.8.2 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 59
5.9 掃描式電子顯微鏡之觀察控制組牙本質試片 60
5.10 硬度計測試結果 60
第六章 討論 61
6.1 離子通透性及選擇性 61
6.1.1 為何在酸性環境下,各種離子都較容易穿透牙本質小管? 61
6.1.2 為何陽離子通透的速率,在酸性及中性時,較陰離子快速? 61
6.1.3 為何帶較多電子數的離子,有較快通過牙本質小管的趨勢? 61
6.1.4 為何鹼性時,各種離子皆難以通過?且其離子選擇性似乎和酸性及中性環境時大有不同? 63
6.1.5 牙本質小管為何在中性時帶負電, 而在酸性及鹼性時,可能是帶何種電荷? 64
6.1.6 為何想了解牙本質小管的選擇性?其意義何在? 64
6.2 GCMS/與不同比例磷酸或磷酸鹽混合製劑, 於牙本質小管內結晶的情形 65
6.2.1 為何GCMS/調整pH值磷酸二氫鉀組別之粉液比,最多做到1:6?為何要調整pH值? 65
6.2.2 為何新配方的含GCMS/磷酸二氫鈉 paste 混合製劑組別之做法改變? 65
6.2.3 為何磷酸二氫鈉和GCMS混合製劑的效果,較磷酸二氫鉀好許多? 66
6.2.4 含鈣中孔洞奈米矽材料/磷酸或磷酸鹽之混合製劑在牙本質內形成良好結晶的條件為何? 66
6.2.5 GCMS的優勢為何?為何一定要加磷酸?其它酸不行嗎? 67
6.2.6 矽可能扮演的角色 69
6.3 為何牙本質試在塗抹GCMS/30%磷酸之混合製劑後,硬度會有些微增加? 69
6.4 鈣離子及磷酸根離子在各pH值下的Ksp 70
第七章 結論 71
第八章 未來研究方向 72
參考文獻 75
附圖 89
附表 141

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