中孔洞二氧化矽奈米粒子於生物醫學上之應用：酵素輸送及標靶治療

Abstract

中孔洞二氧化矽奈米粒子是一個具有多功能型的奈米材料，加上具有高表面積、孔體積、高熱穩定性以及容易在表面進行特殊官能基團修飾等優點的性質，所以廣泛的被應用在催化反應、離子交換等方面，具有相當高的應用價值。近幾年的研究發現，中孔洞二氧化矽奈米粒子具有相當好的生物相容性，以及可以有效率的被生物細胞進行吞噬，被認為是極具潛力的新穎材料，因此有許多的奈米生醫相關應用的研究被大量發表。例如傳遞藥物及基因的載體、醫學造影像以及生物偵測等等。 在本論文第一部分，主要是利用蛋白質工程結合中孔洞二氧化矽奈米粒子的概念，我們是第一個在中孔洞二氧化矽奈米粒子材料上證明變性的蛋白質可以藉由chaperones進行再折疊，成為一個具有生物活性的蛋白質，應用在蛋白質或酵素治療上之新穎研究。 主論文第二部分，在提出一個以奈米粒子作為細胞訊號傳遞的調控者之創新概念，我們設計了一個結合NF-κB p65抗體與HIV 1上TAT胜肽的聰穎中孔洞二氧化矽奈米粒子，藉由抗體辨識及奈米材料的物理性質(尺寸)所造成的立體障礙，直接有效抑制癌細胞內p65的活化更進一步抑制癌細胞的增生。我們同時證明了利用奈米粒子對細胞中核蛋白來進行調控之想法的可行性。

Mesoporous silica nanoparticle (MSN) is one of the promising nanoparticles to serve as a multifunctional vehicle due to its high surface area, uniform pore size, easy functionalization, and biocompatibility. Therefore, it becomes highly suitable for biological applications. In additional to traditional applications of mesoporous silica such as catalysis and chromatography, biomedical applications of MSN such as enzyme immobilization, gene transfection, bioimage and drug delivery agents have recently gained much attention. In the first part of the thesis, the strategy is to combine an engineered protein with MSN to demonstrate the denature protein refolded back into its native form which is probably regulated by chaperones in cells. Our result is the first to report that the denature superoxide dismutase (SOD) enzyme carried by MSN has refolded in the cell and exhibits cellular SOD activity. This novel study can be applied to a protein or enzyme-base therapy in the future. In the second part, we proposed nanoparticles served as nuclear translocation blocker by conjugating antibody to interact with transcriptional activator in the cellular signaling pathways. We first designed a smart functionalized MSN to conjugate nuclear factor-κB (NF-κB) anti-p65 antibody and human immunodeficiency virus 1 (HIV 1) transactivating domain which associated with non-endocytosis and nuclear pore complex (NPC) trapping. This smart MSN particle did exhibit cellular penetration and bind NF-κB p65 when activation, via size hindrance to disrupt the p65 translocation, downstream gene transcription and cell proliferation. Unlike existing research, our results demonstrated a novel idea to use nanoparticles as nuclear translocation blocker for further signal regulation.