Detection and monitoring of the multiple inflammatory responses by photoacoustic molecular imaging using selectively targeted gold nanorods.
Detection and monitoring of the multiple inflammatory responses by photoacoustic molecular imaging using selectively targeted gold nanorods.
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DOI：
10.1364/boe.2.000645
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发表时间：
2011-02-23
影响因子：
3.4
通讯作者：
Kim K
中科院分区：
文献类型：
--
作者：
Ha S;Carson A;Agarwal A;Kotov NA;Kim K
体外细胞实验采用光声分子成像（PMI）技术同时检测和监测细胞间粘附分子-1 （ICAM-1）和e-选择素的上调。将人脐静脉内皮细胞（HUVECs）培养在明胶包膜玻片上，并用炎症因子刺激诱导炎症生物标志物ICAM-1和e -选择素的表达。将长径比（AR）为1:3、吸收中心为715 nm的与抗icam -1抗体结合的金纳米棒（GNRs）和吸收中心为800 nm的与抗e -选择素结合的金纳米棒（GNRs）分别暴露于不同刺激条件下的HUVECs中。利用聚焦高频超声换能器（60 MHz, f/1.5）扫描载玻片细胞顶表面的光声信号。ICAM-1和e -选择素的平均PA信号强度比未受刺激的细胞高约3倍（~10 dB）。荧光成像证实了gnr与受刺激的HUVEC细胞的强结合。将HUVEC细胞暴露于与同型对照抗体结合的gnr中，证实了低水平的非特异性结合。此外，在炎症刺激后0、2、6和24小时，将HUVECs暴露于GNRs偶联的抗icam -1抗体和抗e -选择素抗体。炎症各阶段PA强度与荧光成像和rt-PCR定量比较良好。
In vitro cell experiments have been performed to detect and monitor the upregulation of intercellular adhesion molecule-1 (ICAM-1) and E-selectin simultaneously by photoacoustic molecular imaging (PMI). Human umbilical vein endothelial cells (HUVECs) were grown on gelatin-coated glass slides and stimulated with inflammatory cytokines to induce the expression of the inflammatory biomarkers, ICAM-1 and E-selectin. Gold nanorods (GNRs) of aspect ratio (AR) 1:3 with absorption centered at 715 nm conjugated to anti-ICAM-1 antibody and GNRs of AR 1:3.5 with absorption centered at 800 nm conjugated to anti-E-selectin were exposed to HUVECs with different stimulation conditions. A focused high frequency ultrasonic transducer (60 MHz, f/1.5) was used to scan the photoacoustic (PA) signal over the top surface of the cell containing slides. Averaged PA signal intensity from the stimulated cells was about 3 folds higher (~10 dB) compared to the un-stimulated cells for both ICAM-1 and E-selectin. The strong binding of GNRs to the stimulated HUVEC cells was evidenced by fluorescence imaging. Exposure of HUVEC cells to GNRs conjugated to isotype control antibodies confirms a low level non-specific binding. Also, at 0, 2, 6, and 24 hours after inflammatory stimulation, the HUVECs were exposed to GNRs conjugated anti-ICAM-1 antibody and anti-E-selectin antibody. PA intensity at each stage of inflammation compares well with fluorescence imaging and rt-PCR quantification.