Ag2Se nanoparticles suppress growth of murine esophageal cancer allograft in mice by eliminating Porphyromonas gingivalis

doi:10.12122/j.issn.1673-4254.2025.02.05

Abstract

Abstract:

Objectives To investigate the efficacy of Ag₂Se nanoparticles for eliminating intracellular Porphyromonas gingivalis (P. gingivalis) in esophageal cancer and examine the effect of P. gingivalis clearance on progression of esophageal cancer. Methods Ag₂Se nanoparticles were synthesized via a chemical synthesis method. The effects of Ag₂Se nanoparticles on P. gingivalis viability and colony-forming ability were assessed using fluorescence staining and colony formation assays. In a mouse model bearing subcutaneous murine esophageal cancer cell allograft with P. gingivalis infection, the effect of treatment with Ag₂Se nanoparticles on the abundance of P. gingivalis in the tumor tissues was quantified using RNAscope in situ hybridization and quantitative polymerase chain reaction (qPCR), and the changes in tumor volume were monitored. The biosafety of Ag₂Se nanoparticles was assessed by examining liver and kidney functions and pathological changes in the major organs of the mice. Results Transmission electron microscopy revealed that the synthesized Ag₂Se nanoparticles were uniformly dispersed spherical particles with a diameter around 50 nm. In vitro experiments demonstrated that exposure to Ag₂Se nanoparticles significantly reduced the viability and clonal proliferation capacity of P. gingivalis in a dose-dependent manner. In the tumor-bearing mice, treatment with Ag₂Se nanoparticles significantly reduced the abundance of P. gingivalis in tumor tissues and suppressed tumor cell proliferation. No significant damages to the liver and kidney functions or the major organs were observed in Ag₂Se nanoparticle-treated mice, demonstrating good biocompatibility of Ag₂Se nanoparticles. Conclusion Ag₂Se nanoparticles exhibit significant bactericidal and inhibitory effects against P. gingivalis, and can effectively eliminate intracellular P. gingivalis to suppress the growth of esophageal cancer allograft in mice, suggesting the potential of Ag₂Se nanoparticles in the treatment of esophageal cancer.

Key words: esophageal cancer, Porphyromonas gingivalis, nanomaterials, acid responsive ness

Yali ZHAO, Jiayi LI, Bianli GU, Pan CHEN, Li ZHANG, Xiaoman ZHANG, Pingjuan YANG, Linlin SHI, Shegan GAO. Ag₂Se nanoparticles suppress growth of murine esophageal cancer allograft in mice by eliminating Porphyromonas gingivalis[J]. Journal of Southern Medical University, 2025, 45(2): 245-253.

Figures/Tables 5

Fig.1 Characterization of Ag2Se nanoparticles. A, B: Transmission electron microscopy of Ag2Se nanoparticles and the particle size distribution. C: EDS spectrum of Ag2Se nanoparticles. D: XRD of patterns of Ag2Se nanoparticles. E: Ultraviolet absorption map of Ag2Se nanoparticles at different concentrations. F: Fitting equation of ultraviolet absorption of Ag2Se nanoparticles.

Fig.2 Antibacterial performance evaluation of Ag2Se nanoparticles in vitro. A: P. gingivalis cultures with different treatments for 6 h and inoculated on solid Columbia blood agar plates at 37 ℃ in a anaerobic chamber for 7 days. B: Quantitative map of colony formation. C: CLSM images of P. gingivalis stained with MycoLight™ Green and PI nucleic acid dye (green and red fluorescence represent live and dead bacteria, respectively), at Ag2Se concentration of 80 μg/mL (scale bar=30 µm). D: Quantitative map of live and dead bacterium. E: Bacterial survival after treatment at different drug concentrations (0, 5, 10, 20, 40, 60, 80, 100, 150, 200 μg/mL). F: Effects of different drug concentrations, expressed as logarithmic values (Log10), on bacterial survival. The IC50 value was 70.39 μg/mL. G: Growth curves of P.gingivalis with different treatments. Data are presented as Mean±SD (n=3). *P<0.05, **P<0.01.

Fig.3 Antibacterial evaluation of Ag2Se nanoparticles in tumor tissues in tumor-bearing mice. A: RNAscope in situ hybridization to detect the abundance of P. gingivalis in the tumor tissues (scale bar=25 μm). Red and green fluorescence signals both indicate P. gingivalis. B: Quantification of P. gingivalis DNA in the tumor tissue from different treatment groups using qPCR detection. C: Quantitative analysis of fluorescence intensity. Data are presented as Mean±SD (n=5). *P<0.05, **P<0.01, ***P<0.001.

Fig.4 Effect of Ag2Se nanoparticles on growth of esophageal cancer cells in mice. A: Volume of the dissected tumors in each group. B: Tumor volume curves of the mice with different treatments. C: Tumor weight of the mice treated with saline, MZS and Ag2Se nanoparticles. D: HE staining and Ki67 immunohistochemistry of the tumor sections in each group (scale bar=50 μm). E: Body weight curves of C57 mice with different treatments. F: Quantitative analysis of Ki67 staining in different groups. Data are presented as Mean±SD (n=5). *P<0.05, **P<0.01, ***P<0.001.

Fig.5 Evaluation of biocompatibility of Ag2Se nanoparticles. A: HUVEC viability after incubation with different concentrations of Ag2Se nanoparticles for 24 h. B: Hemolysis assay after different treatments and the hemolysis ratio detected at 540 nm. C-F: ALT, AST, BUN, and CREA of the mice after different treatments. G: HE staining of the heart, liver, spleen, lung, and kidney tissues in different groups (scale bar=50 μm). Data are presented as Mean±SD (n=3).

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Ag₂Se nanoparticles suppress growth of murine esophageal cancer allograft in mice by eliminating Porphyromonas gingivalis

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