請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18605完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 林俊彬 | |
| dc.contributor.author | Kai-Chun Chang | en |
| dc.contributor.author | 張凱鈞 | zh_TW |
| dc.date.accessioned | 2021-06-08T01:14:29Z | - |
| dc.date.copyright | 2014-10-15 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-08-13 | |
| dc.identifier.citation | 1. Hargreaves, K.M., ed. Seltzer And Bender's Dental Pulp. 3rd ed., ed. S. Seltzer and H.E. Goodis. 2002, Quintessence Publishing.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18605 | - |
| dc.description.abstract | 活髓治療的目的在使牙髓保持活性並形成修復性牙本質，維持牙齒正常生理結構及功能。活髓治療發展至今在臨床上仍然沒有理想之材料。磷酸鈣骨水泥(CPCs)在牙科的應用主要是作為骨頭缺損的修補材料，在牙周病、顱顏手術、外傷修復、齒顎矯正等領域都有相關應用。然而，CPCs的硬化時間過長(30-60分)、材料的低機械強度造成易崩解等性質仍舊限制了其在臨床應用的發展。CPCs用於活髓治療需改善初期硬化時間太長的缺點，因此本研究改善CPCs製程使有效縮短初期硬化時間並發展可調控硬化時間之新型磷酸鈣-高分子材料應用於活髓治療。
本研究目的為改良磷酸鈣骨水泥(CPCs)生醫陶瓷材料應用於活髓治療，藉由結合磷酸鈣骨水泥(CSC)發展改良適合活髓治療可引導牙髓組織再生之新型磷酸鈣複合生醫陶瓷材料，並以聚麩胺酸-明膠結合生長因子TGF-β1添加至磷酸鈣-硫酸鈣雙相材料系統，探討與細胞組織交互作用的機轉，以及其誘導牙本質牙髓組織再生的調控模式。本研究分為四大部分，第一部分為材料物理化學性質分析包含晶相結構、固化時間、溫度變化、機械性質、降解測試與鈣離子釋放。第二部分為生物活性指標測試，以SEM與XRD鑑定水合產物結構以及所發展之生醫材料對細胞鈣化 (礦化) 的影響。第三部分為生物相容性分析，以ISO-10993標準評估細胞存活率、細胞毒性。第四部份為建立大動物實驗模型，以犬隻評估所研發之生醫材料與市售材料於牙髓組織再生之差異。 研究結果顯示，藉由添加CSC改善CPC硬化時間長的難以單獨應用於活髓治療缺點，其硬化時間由一個小時降為十五分，符合臨床操作性質，此材料具有良好的生物相容性。此外，所研發之磷酸鈣/硫酸鈣雙相材料能誘導牙本質橋的形成，而以聚麩胺酸-明膠結合生長因子TGF-β1添加至磷酸鈣-硫酸鈣雙相材料系統在一個月即能誘導完整連續之本質橋生成。 | zh_TW |
| dc.description.abstract | Vital pulp therapy is to preserve the pulp vitality as well as to form the reparative dentin for maintaining the normal physical structures and functions of teeth. After years of research and development of the biomaterials, not an ideal material for vital pulp therapy
Bioactive calcium phosphate cements (CPC) have been widely used for repairing bone defects because of their excellent biocompatibility and bioactivity. However, the drawbacks of poor handling properties, low initial mechanical strength, and relatively long setting time limit its application in vital pulp therapy. We synthesized and evaluated the feasibility of biphasic calcium phosphate/sulfate cements for vital pulp therapy. CPC cements were mixed with various amounts of calcium sulfate cements (CSC) to improve the physical, chemical, and mechanical properties. Poly-γ-glumatic acid (γ-PGA) is suitable candidate for drug carrier. And TGF-β1 is well-known for its ability to induce differentiation of dental pulp. The CPC-CSC biphasic biomaterial combinedγ-PGA with TGF-β1 for vital pulp therapy. The purpose of this study is to understand the mechanisms underlying pulp-dentin regeneration and test the efficacy in vivo of our materials for vital pulp therapy. We examined the biphasic biomaterials to study its physicochemical properties, bioactivity, biocompatibility and animal study. The results shows that the initial setting time of CPC-CSH biphasic cement exhibited substantial improvement compared with CPC. The cell viability and cytotoxicity exhibited no harm to the dental pulp cell and animal study presents this CPC-CSH biphasic cement can induce calcified tissue. Based on the results, the developed CPC/CSH biphasic cement has great potential as a material for vital pulp therapy. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T01:14:29Z (GMT). No. of bitstreams: 1 ntu-103-D96422005-1.pdf: 13481010 bytes, checksum: 4709aee31f95edc8b73331e2401ab77f (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 中文摘要------------------------------------------------------------------------------------------i
英文摘要-----------------------------------------------------------------------------------------ii 第一章 導論-------------------------------------------------------------------------------------1 第二章 研究背景與文獻回顧----------------------------------------------------------------2 2.1 保存牙髓活性於牙科醫療之重要性------------------------------------------------2 2.1.1 牙齒之功能與構造------------------------------------------------------------------2 2.1.2 引導牙髓組織再生於臨床治療之意義------------------------------------------2 2.2 活髓治療 (Vital Pulp Therapy)-------------------------------------------------------3 2.2.1活髓治療的目的--------------------------------------------------------------------3 2.2.2 間接覆髓 (Indirect Pulp Capping)----------------------------------------------4 2.2.3 直接覆髓 (Direct Pulp Capping)------------------------------------------------5 2.2.3 斷髓治療 (Pulpotomy)-----------------------------------------------------------5 2.3 現今材料應用於牙髓組織再生之缺點與限制------------------------------------6 2.3.1 Zinc Oxide Eugenol (ZOE)--------------------------------------------------------7 2.3.2 Glass Ionomer (GI)/Resin-Modifed Glass Ionomer (RMGI)-----------------7 2.3.3 Adhesive Systems-------------------------------------------------------------------8 2.3.4 Calcium Hydroxide-----------------------------------------------------------------9 2.3.5 Mineral Trioxide Aggregate (MTA)----------------------------------------------9 2.4 生醫材料誘導牙本質-牙髓組織再生的機轉尚未明瞭-------------------------10 2.5 磷酸/硫酸鈣雙相 (Biphasic) 生醫陶瓷於活髓治療之應用潛力------------10 2.5.1 生醫陶瓷---------------------------------------------------------------------------10 2.5.2 近生物惰性陶瓷 (nearly inert bioceramics)---------------------------------11 2.5.3 生物活性陶瓷 (surface-active bioceramics)---------------------------------12 2.5.4 生物吸收性陶瓷 (resorbable bioceramics)----------------------------------12 2.5.5 磷酸鈣陶瓷與分類--------------------------------------------------------------12 2.5.6 磷酸鈣骨水泥--------------------------------------------------------------------13 2.5.7硫酸鈣骨水泥---------------------------------------------------------------------15 2.6 生長因子對誘導牙髓組織再生扮演重要的角色-------------------------------16 2.6.1 Bone Morphogenetic Proteins (BMPs)----------------------------------------16 2.6.2 TGF-β1於牙髓組織再生之應用潛力---------------------------------------17 2.6.3 以聚麩胺酸-明膠 (γ-PGA/Gelatin) 載體攜帶生長因子------------------17 2.7 生醫材料結合生長因子應用於牙髓組織再生之優勢-------------------------18 2.8 生物相容性對生醫材料研發過程是重要的一環-------------------------------21 第三章 研究目的-----------------------------------------------------------------------------25 第四章 材料與實驗方法--------------------------------------------------------------------26 4.1 磷酸鈣/硫酸鈣雙相材料之備製(CPC-CSC)-------------------------------------27 4.1.1 磷酸鈣骨水泥之備製 (Calcium Phosphate Cements, CPCs)-------------27 4.1.2 磷酸/硫酸鈣雙相材料備製(CPC-CSC)--------------------------------------27 4.1.3 磷酸/硫酸鈣雙相材料添加γ-PGA/Gelating水膠------------------------28 4.2 材料物化性質分析 (Physicochemical Properties)------------------------------28 4.2.1 X光繞射分析（X-ray Diffraction analysis）-------------------------------28 4.2.2 傅立葉轉換紅外線光譜 (FT-IR)---------------------------------------------29 4.2.3硬化時間 (Setting Time)--------------------------------------------------------29 4.2.4 量測材料硬化過程之溫度變化-----------------------------------------------29 4.2.5 酸鹼值評估 (pH Variation)----------------------------------------------------30 4.2.6 抗壓強度測試 (Compressive Strength)--------------------------------------30 4.2.7降解測試---------------------------------------------------------------------------31 4.2.8 鈣離子釋放-----------------------------------------------------------------------31 4.3 體外生物活性測試--------------------------------------------------------------------31 4.3.1 模擬體液溶液SBF之備製-----------------------------------------------------31 4.3.2 表面型態分析-掃描式電子顯微鏡(SEM)觀察-----------------------------33 4.3.3 X光繞射分析（X-ray Diffraction analysis）--------------------------------34 4.4 生物相容性評估-----------------------------------------------------------------------34 4.4.1 人類牙髓細胞 (human pulp cell)之初級培養(primary culture)----------35 4.4.2 萃取液備製-----------------------------------------------------------------------36 4.4.3 細胞存活率實驗（cell viability assay）-------------------------------------37 4.4.4 細胞毒性測試(Lactate Dehydrogenase, LDH)-------------------------------37 4.4.5 生長因子釋放測試--------------------------------------------------------------38 4.5 細胞礦化表現--------------------------------------------------------------------------39 4.5.1 鹼性磷酸酶(Alkaline phosphatase, ALPase)定性染色分析---------------39 4.5.2 細胞基質礦化小體染色(Alizarin Red S staining, ARS)-------------------40 4.5 動物實驗--------------------------------------------------------------------------------40 4.5.1 動物模式---------------------------------------------------------------------------40 4.5.2 電腦斷層(μ-CT)照射-----------------------------------------------------------45 4.5.3 組織學切片標本備置------------------------------------------------------------45 4.5.4 生醫材料與活體組織之形態影響及組織發炎之狀況--------------------45 4.5.5 評估牙髓-牙本質硬組織之生成-----------------------------------------------46 4.6 數據統計分析--------------------------------------------------------------------------47 第五章 結果與討論---------------------------------------------------------------------------48 5.1 CSC含量對CPC-CSC雙相材料的影響------------------------------------------48 5.1.1 XRD繞射分析--------------------------------------------------------------------49 5.1.2 Setting Time-----------------------------------------------------------------------50 5.1.3材料硬化過程之溫度變化------------------------------------------------------51 5.1.4 酸鹼值(pH)變化------------------------------------------------------------------51 5.1.5 機械性質---------------------------------------------------------------------------52 5.1.6材料降解性測試-------------------------------------------------------------------55 5.1.7 SEM 觀察材料水合後之顯微結構--------------------------------------------57 5.1.8 生物相容性------------------------------------------------------------------------59 5.2 含γ-PGA/Gelatin水膠(Gel)之CPC-CSC雙相材料-物理化學性質分析---61 5.2.1 硬化時間---------------------------------------------------------------------------62 5.2.2 水粉比(L/P)對硬化時間之影響-----------------------------------------------63 5.2.3 材料硬化過程之溫度變化------------------------------------------------------64 5.2.4 酸鹼值變化------------------------------------------------------------------------65 5.2.5 機械性質-抗壓強度--------------------------------------------------------------66 5.2.6 材料降解性測試------------------------------------------------------------------67 5.2.7 鈣/磷離子釋放量-----------------------------------------------------------------70 5.3生物活性評估 (Bioactivity)----------------------------------------------------------71 5.3.1 SEM觀察水合產物之表面顯微結構------------------------------------------71 5.3.2水合產物結構分析-XRD--------------------------------------------------------84 5.3.3 氫氧基磷灰石(HAp)轉化率----------------------------------------------------87 5.3.4 生長因子釋放測試---------------------------------------------------------------88 5.4 生物相容性與細胞礦化--------------------------------------------------------------89 5.4.1 Cell Viability – WST-1------------------------------------------------------------89 5.4.2 Cytotoxicity Assay – LDH--------------------------------------------------------90 5.4.3 live and dead stain -----------------------------------------------------------------91 5.4.4鹼性磷酸酶(Alkaline phosphatase, ALPase)定性染色分析----------------94 5.5 動物實驗(Animal Study)-------------------------------------------------------------95 5.5.1電腦斷層掃描(μ-CT)影像分析-------------------------------------------------95 第六章 結論----------------------------------------------------------------------------------101 參考文獻---------------------------------------------------------------------------------------102 圖目錄 圖2-1含TGF-β1之CPC-CSC雙相複合生醫材料引導牙髓組織再生之概念圖------------------------------------------------------------------------------------------------------20 圖4-1 實驗流程圖----------------------------------------------------------------------------26 圖4-2 狗之覆髓過程。(a) 手術部位注射局部射麻藥 (b) 以高速磨牙手機於頰側齒頸部區域車磨第五級窩洞 (c) 使牙髓暴露約0.5~1 mm (d) 已消毒過的小棉球止血 (e) 放置覆髓材料於窩洞內 (f) 填入玻璃離子體封閉窩洞-------43 圖4-3 (a) 定位頸內動脈 (b) 定位頸外靜脈 (c) 靜脈留置針放置於頸內動脈--44 圖5-1-1 1400度高溫燒結焠火後TTCP之XRD 圖譜--------------------------------49 圖5-1-2 CSC比例對CPC-CSC雙相材料初始與最終硬化時間之影響------------50 圖5-1-3 各組材料水合硬化之溫度變化--------------------------------------------------51 圖5-1-4 材料於PBS中pH 值之變化----------------------------------------------------52 圖5-1-5 (A) CSC含量對CPC機械性質-抗壓強度之影響 (B) 各組材料浸泡於SBF隨時間變化之直徑抗拉強度----------------------------------------------------------54 圖5-1-6 (A) 材料浸泡於SBF 二十八天之重量損失率 (B) 材料隨時間之孔隙率變化-------------------------------------------------------------------------------------------56 圖5-1-7 SEM觀察各組材料浸泡於SBF 二十八天後之顯微結構圖 (A) 3CPC/CSC; (B) CPC/CSC; (C) CPC/3CSC------------------------------------------------58 圖5-1-8 生物相容性測試 (A) LDH細胞毒性 (B) 以WST-1評估細胞存活率--------------------------------------------------------------------------------------------------------60 圖5-2-1 Gel對各組材料硬化時間之影響------------------------------------------------63 圖5-2-2不同水粉比(L/P)對材料硬化時間之影響--------------------------------------64 圖5-2-3 各組材料浸泡於PBS一天之pH值變化--------------------------------------66 圖5-2-4 Gel對材料機械性質-抗壓強度之影響-----------------------------------------67 圖5-2-6-1表面裂解圖------------------------------------------------------------------------68 圖5-2-6-2 各組材料之降解程度-重量損失率與視孔隙率 (A) 7CPC/3CSC/Gel; (B) 6CPC/4CSC/Gel---------------------------------------------------------------------------69 圖5-2-7 6CPC/4CSC與6CPC/4CSC/Gel 之鈣離子與磷離子釋放量--------------71 圖5-3-1-1 SEM觀察7CPC/3CSC/Gel浸泡於SBF不同時間點之水合產物表面顯微結構圖 (A)第一天; (B)第三天; (C)第七天; (D)第十四天; (E)第二十八天----------------------------------------------------------------------------------------------------------75 圖 5-3-1-2 SEM觀察6CPC/4CSC/Gel浸泡於SBF不同時間點之水合產物表面顯微結構圖 (A)第一天; (B)5000x; (C)第三天; (D)5000x; (E)第七天; (F)5000x; (G)第十四天; (H)5000x; (I)第二十八天(J)5000x---------------------------------------------80 圖 5-3-1-3 SEM觀察MTA浸泡於SBF不同時間點之水合產物表面顯微結構圖 (A)四個小時; (B)第一天; (C)第三天; (D)第七天; (E)第十四天; (F)第二十八天----------------------------------------------------------------------------------------------------------83 圖5-3-2-1 7CPC/3CSC、7CPC/3CSC/Gel、6CPC/4CSC、6CPC/4CSC/Gel、CPC/CSC與CPC/CSC/Gel水合硬化後之XRD晶相圖譜-----------------------------------------85 圖 5-3-3-2 7CPC/3CSC/Gel 浸泡於SBF水合後之XRD產物晶相分析-----------86 圖 5-3-3-3 6CPC/4CSC/Gel 浸泡於SBF水合後之XRD產物晶相分析-----------87 圖5-3-4 TGF-β1於材料6CPC/4CSC與6CPC/4CSC/Gel中不同時間點之釋放-------------------------------------------------------------------------------------------------------89 圖5-4-1 各組材料以WST-1測試細胞存活率-------------------------------------------90 圖5-4-2 材料對牙髓細胞之LDH細胞毒性測試一天與三天結果------------------91 圖5-4-3-1 材料對牙髓細胞之live and dead stain 作用一天之影像圖--------------92 圖5-4-3-2 材料對牙髓細胞之live and dead stain 作用三天之影像圖--------------93 圖 5-4-4 鹼性磷酸酶定性染色-------------------------------------------------------------94 圖5-5-1 動物實驗μ-CT 影像(A) 6CPC/4CSC活髓治療一個月; (B) 圖圖圖6CPC/4CSC/Gel + TGF-β1活髓治療一個月; (C)氫氧化鈣活髓治療一個月; (D) MTA活髓治療一個月; (E) 6CPC/4CSC活髓治療二個月; (F) 6CPC/4CSC/Gel + TGF-β1活髓治療二個月; (G) 氫氧化鈣活髓治療二個月; (H) MTA活髓治療二個月---------------------------------------------------------------------------------------------100 表次 表2-1 不同鈣磷比之磷酸鈣陶瓷及分類-------------------------------------------------13 表2-2 市售常見磷酸鈣骨水泥系統-------------------------------------------------------15 表4-1 SBF成分表----------------------------------------------------------------------------33 表4-2 人類血漿與SBF所含離子濃度---------------------------------------------------33 表4-3 組織學評評分標準-------------------------------------------------------------------46 表4-4 Hard Tissue Bridge生成評估表----------------------------------------------------47 表5-1 不同水膠比例之固化時間----------------------------------------------------------47 表5-2 各組材料硬化過程中之溫度差----------------------------------------------------62 表5-3 各晶相之JCPDF標準圖譜，繞射平面強度與角度表------------------------65 表5-4 μ-CT 影像硬組織生成評估-----------------------------------------------------100 | |
| dc.language.iso | zh-TW | |
| dc.title | 研發含TGF-β1之磷酸鈣/硫酸鈣雙相水泥於活髓治療之研究 - 材料物化性質、生物活性、生物相容性及動物實驗 | zh_TW |
| dc.title | Development of Calcium Phosphate/Calcium Sulfate Biphasic Cements with TGF-β1 for Vital Pulp Therapy - Physicochemical Properties, Bioactivity, Biocompatibility and Animal Study | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.coadvisor | 林?輝 | |
| dc.contributor.oralexamcommittee | 鄭景暉,章浩宏,姜昱至,林弘萍,鄧麗珍 | |
| dc.subject.keyword | 磷酸鈣骨水泥,硫酸鈣,活髓治療,生物相容性,TGF-β1, | zh_TW |
| dc.subject.keyword | calcium phosphate cements,calcium sulfate,vital pulp therapy,TGF-β1, | en |
| dc.relation.page | 110 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2014-08-13 | |
| dc.contributor.author-college | 牙醫專業學院 | zh_TW |
| dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
| 顯示於系所單位： | 臨床牙醫學研究所 | |
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