摻銀之氫氧基磷灰石奈米粒子做為肺癌的診斷學治療

Development of Ag-doped Hydroxyapatite Nanoparticle as Theranostics Agent for Lung Cancer

關金東(Indra Oliver Candra Hadyiswanto)

指導教授 ： 林峯輝

國立臺灣大學/工學院/醫學工程學研究所/碩士(2013年)

https://doi.org/10.6342/NTU.2013.03109

摘要

肺癌是世界上最致命的疾病之一，每年造成至少1.3萬人死亡。肺癌之生物醫學影像目前仍無法精確的成像。另一方面，肺癌的治療方法只能針對於較小範圍的區域進行治療，並有許多副作用。許多研究證實了同時具有診斷及治療功能之奈米藥物是具有潛力成為未來治療癌症的工具。而目前許多研究皆已利用同時具有診斷及治療之奈米藥物成功改善了生醫影像之靈敏度及增進治療癌症之效用。在本研究中，我們將製備摻銀之氫氧基磷灰石奈米粒子，並使之成為治療肺癌上之電腦斷層掃描(CT scan)顯影劑以及化療藥物。
在本研究中，我們利用共沉澱法製備摻銀之氫氧基磷灰石奈米粒子。在合成過程中，不同濃度之銀離子被摻入氫氧基磷灰石當中。在材料定性分析實驗當中使用X光繞射儀(XRD)譜進行鑑定晶體結構並計算其晶體參數，而藉由掃描式電子顯微鏡(SEM)及穿透式電子顯微鏡(TEM)進行確認材料之粒徑大小及形態，除此之外，高解析度之穿透式電子顯微鏡影像與電子繞射圖譜可分析其晶體結構，於粒徑分析中，藉由粒徑分析儀(DLS)可以確認材料於水相中之粒徑大小，而傅立葉轉換紅外光譜儀(FTIR)可分析材料表面之官能基。為了確認銀離子於氫氧基磷灰石內之數量，我們使用了ICP-MS來進行其元素分析，斷層掃描成像實驗是為了得知材料之X射線衰減係數。在細胞實驗當中，使用WST-1、LDH 及Live/Death細胞染色實驗分別進行摻銀之氫氧基磷灰石奈米粒子之癌細胞毒殺測試。
由實驗結果得知，我們成功的使用共沉澱法合成了摻銀之氫氧基磷灰石奈米粒子。 而由XRD, FTIR, SEM和TE實驗結果可以得知，相較於氫氧機磷灰石奈米粒子，摻銀之氫氧基磷灰石奈米粒子在於大小、形狀、表面之官能基及形態上並無明顯變化，但其繞射峰之位置有些微的改變且其晶體參數較高，證實了其銀離子確實可以置換出鈣離子， ICP-MS的結果可以定量出銀離子在於氫氧基磷灰石晶格內濃度。此外，Micro-CT成像與癌細胞毒殺測試結果證實了摻銀之氫氧基灰石奈米粒子同時具有顯影以及治療癌症的功能，因此，摻銀之氫氧基灰石奈米粒子相當具有潛力成為肺癌治療上之一種theranostic agent。

關鍵字

肺癌 ； 氫氧基磷灰石 ； 銀 ； 化療 ； CT掃描 ； 顯影劑 ； 治療診斷

並列摘要

Lung cancer is one of the deadliest diseases in the world, causing at least 1.3 million people to die every year. Recent biomedical imaging modalities for lung cancer sometimes fail to carry out a precisely correct action since each of them lacks particular attributes. On the other hand, treatments for lung cancers are only effective to a small extent and have numerous side effects. Theranostics nanomedicine has been studied, proposed and proven to be the future tool in cancer treatment. Nanoparticles were successfully employed in previous studies so as to improve biomedical imaging sensitivity and / or cancer treatment efficacy. In this study, we are going to use silver-doped hydroxyapatite nanoparticle as a theranostics agent that acts as both CT contrast agent as well as chemotherapeutic agent for lung cancer management.
The nanoparticles were prepared through co-precipitation process. During the process, hydroxyapatite was doped with various amount of silver. The crystal structure and lattice parameters were evaluated by XRD. The morphology and size of the nanoparticles were evaluated by using SEM and DLS. FTIR spectroscopy was performed to determine functional groups in the synthesized particles. TEM experiment was also carried out to examine crystallite size, shape and structure. In order to track and quantify silver ion inside silver-doped hydroxyapatite, ICP-MS experiment was carried out. Micro-CT scan imaging experiment was conducted in order to investigate the X-ray attenuation coefficient of the synthesized particles. After material analyses, in-vitro experiments through WST-1, LDH and Live/Dead cell viability assays were performed to determine the effects of silver hydroxyapatite on lung cancer cell.
Results showed that the AgHAP nanoparticles were successfully synthesized through co-precipitation method. XRD, FTIR, SEM and TEM results revealed that the substitution of calcium by silver in HAP did not change the size, functional groups, shape and morphology of the nanoparticles. However, substitution was proved through change in lattice parameters and shift in XRD spectra. In addition, ICP-MS results proved that Ag ion in AgHAP could be quantified. Micro-CT imaging results and in-vitro experiment results both indicated that AgHAP nanoparticles have the potential as theranostic agent for lung cancer management.

並列關鍵字

lung cancer ； hydroxyapatite ； silver ； chemotherapy ； CT scan ； contrast agent ； theranostics

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