表面修飾玻尿酸金奈米粒子之合成方法及生物活性上之應用

Synthesis and Biological Activities of Gold Nanoparticles Stabilized with Hyaluronic Acid

呂維哲(Wei-Che Lu)

指導教授 ： 施養佳 ； 張培均 ； 鄧金培

亞洲大學/健康學院/生物科技學系碩士班/碩士(2010年)

摘要

金奈米粒子是一種穩定且具抗氧化功能的奈米材料，並且生物相容性極佳，因為容易與蛋白質結合，如抗體、酵素或細胞激素等，常被用於醫學檢測與藥物釋放，甚至有研究指出，金奈米粒子還可以被用來治療關節炎，相關應用領域廣闊。玻尿酸為脊椎動物普遍存在的生物性高分子，原料來源有動物組織萃取及微生物發酵。生理功能包含調控細胞的遷移與生長及血管新生作用等，在結構上因為富含OH基團及帶大量負電所以有強效的保濕性並且可以降低活性氧化物(ROS)對細胞造成傷害，其性質隨著分子量的不同也有所改變，因此普遍應用於生物醫學、藥物遞送系統及化妝保養品方面，近年來研究顯示利用腫瘤細胞附近會大量表現玻尿酸受器及玻尿酸分解酶的特性，做為檢測的分子生物標誌。本研究之主目的為合成玻尿酸金奈米粒子，結合兩者之特性，建立生物相容性指標，並開發其生物活性應用上之潛力。本研究概分為三個部分進行，第一部分為合成玻尿酸金奈米粒子，並探討酸鹼值、溫度、還原劑濃度及玻尿酸濃度等合成條件和最佳化流程，並比較其物理性質包含穩定性、粒徑、光學特性和玻尿酸黏度之差異。第二部分為評估玻尿酸金奈米粒子之生物相容性。鑑定玻尿酸金奈米粒子是否具細胞增生活性或毒性，探討玻尿酸金奈米粒子的生物相容性和無細胞毒性最低劑量。最後評估玻尿酸金奈米粒子做為抗氧化劑之可行性，藉由清除自由基的能力驗證玻尿酸金奈米粒子是否具抗氧化能力。本研究完成開發一個具高效率利用0.2%玻尿酸代替PEG並以0.02M NaBH4合成平均粒徑為3.68nm之金奈米粒子的方法，並且得到在不同合成狀態下所造成差異的條件，此外在經過體外細胞存活率測試後，證明了玻尿酸合成金奈米粒子並無細胞毒性，其無毒性最低劑量大於100ug/mL，此結果也證明還原劑對細胞並無明顯毒性。而抗氧化活性方面之結果，雖然顯示玻尿酸金奈米粒子具DPPH自由基清除率52.79%。在96孔盤形成玻尿酸基質表面會抑制癌細胞 (MCF-7) 貼附，達到43.49% 抑制率。不論是基礎科學研究或生醫材料的開發，都具有高度的產業化及競爭性。

關鍵字

金奈米粒子 ； 玻尿酸 ； 生物活性 ； 抗氧化活性

並列摘要

Hyaluronic acid (HA) is a natural polymer used in biomedical and cosmetic applications, and which has great water-holding capacity and high viscosity. HA also contributes significantly to various cellular processes such as adhesion, migration, and proliferation, as well to assist in the control of angiogenesis. Gold nanoparticles with the great stability present fascinating antioxidant activity and biocompatibility. HA and gold nanoparticles are the potential biomaterials to apply in medicine, drug delivery and cosmetics. In this study, gold-hyaluronic acid nanoparticles were prepared by chemical reduction method, and various particle sizes have been synthesized by the different concentration of the reducing agent. The absorption spectra of gold-hyaluronic acid nanoparticles measured by UV-visible spectrophotometer were observed near 520 nm. Their particle sizes and shapes were investigated by transmission electronmicroscope (TEM), which indicated the formation of uniform spherical particles with well dispersion (2-6nm). Therefore, the synthesis of size-controlled gold-hyaluronic acid nanoparticles could be achieved by chemical reduction methods, and the resultant of gold-hyaluronic acid nanoparticles is very stable and showed non-toxicity to NIH3T3 cell. The DPPH scavenging capacity is 52.79%, suggested as a candidate molecule for antioxidant. Pre-coating HA treat to tumor cell line (MCF7) showed anti-adhesion activity and inhibition rat is 43.49%. the results suggest gold-hyaluronic acid nanoparticles have potential in medicine applications, such as biomaterial and anticancer treatment.

並列關鍵字

gold nanoparticle ； Hyaluronic acid ； Biological Activities ； antioxidant

參考文獻

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