Sanità Gennaro, Armanetti Paolo, Silvestri Brigida, Carrese Barbara, Calì Gaetano, Pota Giulio, Pezzella Alessandro, d'Ischia Marco, Luciani Giuseppina, Menichetti Luca, Lamberti Annalisa
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
Institute of Clinical Physiology, National Research Council, Pisa, Italy.
Front Bioeng Biotechnol. 2020 Jul 8;8:765. doi: 10.3389/fbioe.2020.00765. eCollection 2020.
Bioconjugation of a recently developed photoacoustic nanoprobe, based on silica-templated eumelanin-silver hybrid nanoparticles (MelaSil_Ag-NPs), with human serum albumin (HSA) is disclosed herein as an efficient and practical strategy to improve photostability and to perform SPARC mediated internalization in breast cancer cells. Modification of NPs with HSA induced a slight viability decrease in breast cancer cells (HS578T) and normal breast cells (MCF10a) when incubated with HSA-NPs up to 100 μg/mL concentration for 72 h and a complete suppression of hemotoxicity for long incubation times. Uptake experiments with MelaSil_Ag-HSA NPs indicated very high and selective internalization via SPARC in HS578T (SPARC positive cells) but not in MCF10a (SPARC negative cells), as evaluated by using endocytosis inhibitors. The binding of SPARC to HSA was confirmed by Co-IP and Dot-blot assays. Additional studies were performed to analyze the interaction of MelaSil_Ag-HSA NPs with protein corona. Data showed a dramatic diminution of interacting proteins in HSA conjugated NPs compared to bare NPs. HSA-coated MelaSil_Ag-NPs are thus disclosed as a novel functional nanohybrid for potential photoacoustic imaging applications.
本文公开了一种基于二氧化硅模板化真黑素 - 银杂化纳米颗粒（MelaSil_Ag - NPs）的新型光声纳米探针与人血清白蛋白（HSA）的生物共轭，这是一种提高光稳定性并在乳腺癌细胞中实现SPARC介导的内化的有效且实用的策略。当与浓度高达100μg/mL的HSA - NPs孵育72小时时，用HSA修饰纳米颗粒会导致乳腺癌细胞（HS578T）和正常乳腺细胞（MCF10a）的活力略有下降，并且长时间孵育可完全抑制血液毒性。通过使用内吞作用抑制剂评估，MelaSil_Ag - HSA NPs的摄取实验表明，在HS578T（SPARC阳性细胞）中通过SPARC实现了非常高的选择性内化，但在MCF10a（SPARC阴性细胞）中没有。通过Co - IP和斑点印迹分析证实了SPARC与HSA的结合。还进行了其他研究以分析MelaSil_Ag - HSA NPs与蛋白质冠的相互作用。数据显示，与裸纳米颗粒相比，HSA共轭纳米颗粒中相互作用蛋白显著减少。因此，HSA包覆的MelaSil_Ag - NPs被公开为一种用于潜在光声成像应用的新型功能性纳米杂化物。
