Preparation of fluorescent nanoprobes based on aggregation-induced emission and their application in the diagnosis and treatment of oral cancer
10.3760/cma.j.cn121382-20250723-00056
- VernacularTitle:基于聚集诱导发光的荧光纳米探针的制备及其在口腔癌诊疗中的应用
- Author:
Yanze WANG
1
;
Ruixin NIE
;
Guanhua WANG
;
Xiaoli LIAN
;
Yingbin YAN
;
Xiaoyan ZHANG
Author Information
1. 天津市口腔医院中心实验室 天津市口腔功能重建重点实验室 南开大学医学院，天津 300041
- Keywords:
Aggregation-induced emission;
Fluorescent nanoprobes;
Oral cancer;
Precision tumor diagnosis and therapy
- From:
International Journal of Biomedical Engineering
2025;48(5):443-453
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To develop fluorescent nanoprobes with aggregation-induced emission characteristics and to systematically evaluate their optical properties, biosafety, anti-tumor activity, and imaging capability, thereby assessing their potential for early precision diagnosis and treatment of oral cancer in mice.Methods:Control probes (PEG@TPD) were prepared by encapsulating ( E)-4-(2-(4′-(1-phenyl-2,2-bis(4-methoxyphenyl)vinyl)biphenyl-4-yl)vinyl)-4-(dicyanomethylene)-4 H-chromene (TPD) using 1,2-distearoyl- SN-glycerol-3-phosphoethanolamine- N-polyethylene glycol 2000-maleimide as the carrier. Fluorescent nanoprobes (GE11-PEG@TPD) were subsequently fabricated by surface modification with the targeting GE11 peptide. The morphology and particle size of the nanoprobes were characterized by transmission electron microscopy and dynamic light scattering. The optical properties of the nanoprobes were analyzed using ultraviolet-visible spectrophotometry and fluorescence spectrophotometry. Mouse squamous carcinoma SCC-7 cells were randomly divided into six groups by the random number table method. The PBS, PEG@TPD, and GE11-PEG@TPD groups were not treated with light, while the PBS+L, PEG@TPD+L, and GE11-PEG@TPD+L groups were exposed to white light (25 W/cm 2, 10 min) at a nanoprobe concentration of 20 μg/ml (based on TPD concentration). Cell survival rate was assessed by the cell counting kit-8 assay. Cellular uptake, intracellular reactive oxygen species levels, and cytotoxicity were evaluated using laser scanning confocal microscopy. The apoptosis rate was evaluated by cell apoptosis assay. Twelve 6-week-old female C3H/HeN mice were randomly divided into two groups: PEG@TPD-1 group and GE11-PEG@TPD-1 group, with 6 mice in each group. Subcutaneous oral cancer models were established by injecting SCC-7 cell suspensions into the dorsal region of mice in two groups. Each mouse was intravenously administered 200 μl of PEG@TPD or GE11-PEG@TPD solution (1 mg/ml, based on TPD concentration). Tumor boundaries and scope were visualized using a small animal in vivo imaging system. At the optimal imaging time point, three mice from each group were euthanized, and major organs and tumor tissues were collected to measure probe accumulation. Statistical comparisons between two groups were performed using independent samples t-tests, while one-way or two-way analysis of variance was applied for multiple group comparisons. Results:Both PEG@TPD and GE11-PEG@TPD exhibited a relatively regular sphere, with average particle sizes of (92.76±8.80 and 117.50±6.40) nm, respectively. PEG@TPD showed two obvious absorption peaks at 352 and 444 nm. GE11 peptide showed a polypeptide characteristic absorption peak at 280 nm, GE11-PEG@TPD showed three characteristic absorption peaks at 280, 352 and 444 nm. Under dark conditions, cell survival rate remained above 80% even at a concentration of 160 μg/ml. After light irradiation, cell survival rate in the PEG@TPD+L group at 20 and 40 μg/ml [(68.2±5.2)% and (48.6±7.1)%] were higher than those in the GE11-PEG@TPD+L group [(55.0±2.8)% and (30.0±9.2)%], with statistically significant differences ( P<0.05, 0.01). At incubation time points of 2, 4, and 6 h, the relative fluorescence intensity of the GE11-PEG@TPD group (119.4±10.2, 192.9±14.2, and 234.1±4.8) were higher than those of the PEG@TPD group (98.6±7.5, 163.8±3.1, 204.6±11.2), with statistically significant differences (all P<0.05). The relative fluorescence intensity of the PEG@TPD+L and GE11-PEG@TPD+L group (68.5±4.7 and 86.8±10.0) were higher than those in the PBS, PEG@TPD, GE11-PEG@TPD, and PBS+L groups (6.1±8.0, 7.6±1.8, 4.7±4.2 and 21.1±7.6), with statistically significant differences (all P<0.01). And the difference between the GE11-PEG@TPD+L and PEG@TPD+L groups was also statistically significant ( P<0.05). Viable cell proportions in the PBS, PEG@TPD, GE11-PEG@TPD, and PBS+L groups all exceeded 95.0%, while those in the PEG@TPD+L and GE11-PEG@TPD+L groups decreased to (11.1±3.7)% and (4.3±1.1)%, respectively, with a statistically significant difference between them ( P<0.05). The apoptotic cell proportions in the PEG@TPD+L and GE11-PEG@TPD+L groups [(40.5±4.3)% and (55.3±7.4)%] were higher than those in the PBS, PEG@TPD, GE11-PEG@TPD, and PBS+L groups [(27.3±2.0)%, (28.2±1.9)%, (28.6±1.2)%, and (29.7±3.0)%], with statistically significant differences ( P<0.05, 0.01). Moreover, the difference between the GE11-PEG@TPD+L and the PEG@TPD+L groups was also statistically significant ( P<0.01). The mean fluorescence intensities of the GE11-PEG@TPD-1 group at 1, 3, 5, 8, and 24 h, as well as in ex vivo tumor tissues[(5.2±0.8, 5.9±0.7, 6.6±1.0, 7.9±0.6, 7.8±0.7 and 20.6±3.5)×10 6 p/s/cm 2/sr] were all higher than those in the PEG@TPD-1 group [(3.2±0.7, 4.2±0.7, 4.6±0.9, 5.1±0.9, 4.7±0.9 and 14.2±1.8)×10 6 p/s/cm 2/sr], with statistically significant differences ( P<0.05, 0.01). Conclusions:The fluorescent nanoprobes exhibit uniform particle size, high photostability, and good biocompatibility. They demonstrate significant tumor-killing effects at the cellular level and possess tumor-targeting capability in vivo, showing promising application potential for the early precision diagnosis and treatment of oral cancer.
