臺灣博碩士論文加值系統

牙周炎是盛行率極高的慢性疾病，但大多數的人不會積極去做治療。 牙周炎最初的症狀有: 發炎，流血，牙齦壓痛，最後會導致齒槽骨地吸收造成牙齒的脫落。從材料特性而言，奈米金棒可吸收近紅外光，而近紅外光能有效穿透生物組織。在先前的實驗中我們看到了藉由近紅外光照射奈米金棒產生的局部高溫能殺死致病細菌。在本篇研究中，我們將評估以金奈米柱體粒子為基礎之牙周病治療效果。我們已經透過一種具有可依一定比例延展和放大規模生產的金奈米金棒合成方法，而我們合成的奈米金棒在縱橫比約3.83，對於近紅外光範圍有最大的吸收效果。此奈米金棒透過2-mercaptoethyl amine的修飾能使奈米金棒表面具有安機官能基，而我們設計的奈米金棒也添加聚乙二醇，其目的是為了改善奈米金棒的穩定性以及叫良好的生物相容性。並藉以和專門標靶牙周炎發展中扮演重要角色的Porphyromonas gingivalis(P.g)之外膜蛋白的抗體做一共價結合。使我們的奈米金棒的對 Porphyromonas gingivalis能透過生物標靶與近紅外光照射同步產生高溫與光動力治療。因此，我們在每一個接上抗體之奈米金棒接上四個P.g菌，再將此金棒透過近紅外光雷射暴露，透過其他實驗方法來證實，我們是第一個使用奈米金棒結合P.g菌透過此高熱治療可以有效地將Porphyromonas gingivalis菌裂解。而這也是以標把性金奈米柱體結合光動力與熱聊或許能做為治療牙周病的一種新方法，並藉以提升此一疾病之預防與治療效果。

Periodontitis is a disease that extremely prevalent but often goes untreated. Periodontitis can ultimately lead to the loss of bone and tissues that support the teeth and an inevitable loss of teeth. Gold nanorod conjugates harbor great potential to combat this disease via its photothermal property and biocompatibility. Gold nanorods have optical absorption around the near infrared range that can penetrate deeply through biological tissue. Previous experiments have shown that gold nanorods are capable of killing pathogenic bacteria through near-infrared (NIR) laser irradiation through hyperthermia. In this study, we will evaluate the effects of gold nanoparticle-based therapeutic probes in the treatment of periodontal disease. We have synthesized our gold nanorods via a seed-and-growth method that allows for proportional scalability and mass production of gold nanorods with an aspect ratio of approximately 3.83 and maximum absorption in the NIR range. Our gold nanorods were modified with 2-mercaptoethyl amine in order to obtain surface amine groups. Additionally, the gold nanorods were conjugated with polyethylene glycol (PEG) to improve the stability and biocompatibility of our nanorod. The previously attached amine groups were then used to covalently attach antibodies that specifically target the outer membrane protein of Porphyromonas gingivalis, a pathogenic bacteria that plays a prominent role in the development of periodontitis. Enabling specific targeting of our modified gold nanorod allows for targeted hyperthermia against P. gingivalis. We conjugated around four P. gingivalis targeting antibodies to each gold nanorod. Through use of our targeting gold nanorods, combined with NIR laser exposure, we demonstrate for the first time in literature that targeted hyperthermia is an effective method for lysing P. gingivalis bacteria. The development of gold nanoparticle-based therapeutic probes targeting periodontal disease can lead to new ways of treating, preventing, and overcoming the consequences of periodontal disease.

English Abstract........................................................................................................I
Chinese Abstract......................................................................................................II
Acknowledgements.................................................................................................IV
Table of Contents.....................................................................................................V
List of Tables........................................................................................................VIII
List of Figures.........................................................................................................IX
Abbreviations............................................................................................................X
1. Introduction.........................................................................................................1
1.1 Periodontitis........................................................................................................1
1.1.1 Periodontitis Etiology....................................................................................1
1.2 Prevalence of Periodontitis................................................................................2
1.3 Risk Factors........................................................................................................2
1.3.1 Cardiovascular Heart Disease......................................................................3
1.3.2 Low Birth Weight...........................................................................................3
1.3.3 Socioeconomic Factors...................................................................................3
1.3.4 Age-Related Factors.......................................................................................4
1.3.5 Diabetes...........................................................................................................4
1.3.6 Smoking...........................................................................................................5
1.4 Treatments for Periodontitis..............................................................................5
1.4.1 Nonsurgical.....................................................................................................5
1.4.2 Surgical...........................................................................................................6
1.4.3 Bone Regeneration.........................................................................................6
1.5 Nanotechnology...................................................................................................6
1.5.1 Nanotherapy...................................................................................................7
1.5.2 Nanoparticle Properties.................................................................................7
1.5.3 Targeting Therapeutic Nanoparticles..........................................................8
1.5.4 Photothermal Therapy..................................................................................8
1.5.5 Rationale.........................................................................................................9
2. Materials and Methods.....................................................................................10
2.1 Preparation of CTAB-capped Gold Nanorods...............................................10
2.1.1 Seed Solution Synthesis.................................................................................10
2.1.2 Growth Solution Synthesis............................................................................10
2.1.3 Gold Nanorod Collection...............................................................................10
2.1.4 Gold Nanorod Analysis..................................................................................11
2.2 Amine-Modified Gold Nanorods.....................................................................11
2.2.1 Preparation of Amine-Modified Gold Nanorods........................................11
2.2.2 Analysis of Amine-Modified Gold Nanorods...............................................11
2.3 Polyethylene Glycol Modification....................................................................12
2.3.1 Addition of Polyethylene Glycol...................................................................12
2.3.2 Analysis of PEGylated Gold Nanorods........................................................12
2.4 Antibody Attachment.......................................................................................12
2.4.1 Attachment of P. gingivalis Targeting Antibody.........................................12
2.4.2 Analysis of Antibody Conjugation................................................................13
2.5 Treatment of P. gingivalis with Therapeutic Gold Nanorods.......................13
2.5.1 Incubation of P. gingivalis with Targeting Gold Nanorods........................13
2.5.2 Analysis of Nanorod Binding via TEM........................................................13
2.5.3 Analysis of Nanorod Efficacy via Serial Dilution........................................14
2.5.4 IC50 Determination.........................................................................................14
3. Results..................................................................................................................15
3.1 Synthesis of Gold Nanorods.............................................................................15
3.2 Modification of Gold Nanorods to Have Surface Amine Groups.................15
3.3 PEGylation of Gold Nanorods.........................................................................16
3.4 Antibody Conjugation to Gold Nanorods.......................................................16
3.5 Therapeutic Effect of P. gingivalis Antibody Conjugated Gold Nanorods..17
4. Discussion.............................................................................................................19
5. Summary..............................................................................................................23
6. References............................................................................................................24
7. Figures and Tables..............................................................................................31
8. Personal Profile...................................................................................................47

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