中孔洞二氧化矽奈米材料的特性與其蛋白質冠冕在奈米材料與生物系統上交互作用的影響

Abstract

中孔洞二氧化矽奈米材料在奈米生醫領域的應用上具有多功能的發展性，例如：標靶治療、顯影、追蹤等，然而當具有特定功能的奈米粒子進入生物系統時，大量的蛋白質吸附到奈米粒子表面，形成蛋白質冠冕，導致應用效果不如預期，因此了解這層介於奈米材料與生物體之間的蛋白質冠冕將有助於奈米生醫應用上的發展。 在我們的研究中，我們首先透過液相層析串聯質譜儀、熱重分析儀、動態光散色分析儀來鑑定奈米粒子在老鼠或人類血漿中蛋白質冠冕的成、吸附量多寡、以及表面電位的變化，並透過 ClueGo 和 IPA 進行代謝路徑分析，接著觀察蛋白質冠冕在 in vitro 及 in vivo 實驗結果上的影響，而後綜合上述實驗分析結果選出特定蛋白質：免疫球蛋白 G、富含組氨酸的糖蛋白、間-α-胰蛋白酶抑製劑重鏈 H4，運用這三種蛋白質將奈米粒子進行表面修飾，並注入進帶有腫瘤的動物模型中觀察蛋白質冠冕改變後奈米材料在生物系統內的高滲透長滯留效應及血液循環效果，研究結果顯示將材料進行蛋白質的表面修飾無法輕易的改善材料在生物體的應用性。

Mesoporous Silica Nanoparticles (MSNs) have been developed for many biomedical purposes, such as targeted therapy and diagnostic system. However, once nanoparticles are exposed to the biological system, protein corona formed around the nanoparticles can result in unexpected outcome in terms of therapeutic efficiency, bio-distribution and pharmacokinetics. Therefore, understanding the composition as well as functionality of protein corona is crucial for nano-medicine development. In our study, we use LC-MS/MS, SDS-PAGE, DLS and Zeta potential measurements to identify the protein corona after the in vitro incubation of nanoparticles with human plasma and in vivo injection of nanoparticles into the mice. Additional bio information were revealed by ClueGo and Ingenuity Pathway Analysis (IPA). According to the obtained information, we selected and conjugated three of the most prominent proteins including Immunoglobulin G, Histidine-rich glycoprotein and Inter-alpha trypsin inhibitor H4, to MSNs and exam their bio-distribution. Our results indicate the high tumor-targeting efficiency could be achieved more simply by reducing protein adsorption but not manipulating protein type on the surface of MSNs.