Demirci Hakan, Wang Yichun, Li Qiaochu, Lin Cheng-Mao, Kotov Nicholas A, Grisolia Anna Beatriz Diniz, Guo Jay L
Department of Ophthalmology and Visual Sciences, Kellogg Eye Center, University of Michigan Ann Arbor, MI, USA.
Biointerfaces Institute, University of Michigan Ann Arbor, MI, USA.
J Ophthalmic Vis Res. 2020 Oct 25;15(4):446-452. doi: 10.18502/jovr.v15i4.7778. eCollection 2020 Oct-Dec.
To evaluate the penetration of carbon nanotubes (CNTs) throughout retinoblastoma in a transgenic mice model.
CNTs functionalized with fluorescein isothiocyanate and targeting ligands biotin (CTN-FITC-Bio, 0.5mg/ml), or folic acid (CNT-FITC-FA, 0.5mg/ml) were injected into the vitreous of one eye of LH T transgenic mice. Other eye did not receive any injection and was used as control. Three mice were sacrificed at days 1, 2, and 3. Eyes were enucleated and stained with 4,6-diamidino-2-phenylindole. The sections were imaged by fluorescent microscope. The images were transformed into grey-scale in MATLAB for intensity analysis. Background intensity was normalized by marking squares outside the eyeball and using the mean intensity of these squares. Fluorescent intensity (FI) for each image was measured by calculating the intensity of a same-sized square within retinoblastoma.
Nine eyes of nine mice were included in each CNT-FITC-Bio and CNT-FITC-FA groups. The mean FI in CNT-FITC-Bio was 52.08 6.33, 53.62 9.00, and 65.54 5.14 in days 1, 2, and 3, respectively. The mean FI in CNT-FITC-FA was 50.28 7.37, 59.21 6.43, and 58.38 2.32 on days 1, 2, and 3, respectively. FI was significantly higher in eyes injected with CNT-FITC-Bio and CNT-FITC-FA compared to the control eyes ( = 0.02). There was no difference in FI between eyes with CNT-FITC-Bio and CNT-FITC-FA, and FI remained stable on days 1-3 in CNT-FITC-Bio, CNT-FITC-FA, and control eyes ( 0.05).
We observed higher FI in eyes with CNT-FITC-Bio and CNT-FITC-FA compared to control eyes, showing penetration of CNTs throughout retinoblastoma. CNTs can be a carrier candidate for imaging or therapeutic purposes in retinoblastoma.
在转基因小鼠模型中评估碳纳米管（CNTs）在视网膜母细胞瘤中的渗透情况。
将用异硫氰酸荧光素和靶向配体生物素（CTN-FITC-Bio，0.5mg/ml）或叶酸（CNT-FITC-FA，0.5mg/ml）功能化的碳纳米管注入LHT转基因小鼠一只眼睛的玻璃体中。另一只眼睛不进行任何注射，用作对照。在第1、2和3天处死3只小鼠。摘除眼球并用4,6-二脒基-2-苯基吲哚染色。切片用荧光显微镜成像。图像在MATLAB中转换为灰度图进行强度分析。通过标记眼球外的方块并使用这些方块的平均强度对背景强度进行归一化。通过计算视网膜母细胞瘤内相同大小方块的强度来测量每个图像的荧光强度（FI）。
CNT-FITC-Bio组和CNT-FITC-FA组各纳入9只小鼠的9只眼睛。CNT-FITC-Bio组在第1、2和3天的平均FI分别为52.08±6.33、53.62±9.00和65.54±5.14。CNT-FITC-FA组在第1、2和3天的平均FI分别为50.28±7.37、59.21±6.43和58.38±2.32。与对照眼相比，注射CNT-FITC-Bio和CNT-FITC-FA的眼睛的FI显著更高（P = 0.02）。CNT-FITC-Bio和CNT-FITC-FA的眼睛之间的FI没有差异，并且在第1 - 3天，CNT-FITC-Bio、CNT-FITC-FA和对照眼的FI保持稳定（P>0.05）。
我们观察到与对照眼相比，注射CNT-FITC-Bio和CNT-FITC-FA的眼睛具有更高的FI，表明碳纳米管在视网膜母细胞瘤中具有渗透性。碳纳米管可以作为视网膜母细胞瘤成像或治疗目的的载体候选物。
