1：Experimental Design and Method Selection:

The study involved synthesizing Cu-tryptone complex nanoparticles (Cu-Try NPs) via a simple green method, loading them with methylene blue (MB) to form Cu-Try/MB NPs, and evaluating their ability to consume GSH and enhance photodynamic therapy (PDT) through in vitro and in vivo experiments. Theoretical models include redox reactions between Cu-Try NPs and GSH, and PDT mechanisms involving ROS generation.

2：Sample Selection and Data Sources:

HeLa cells and U14 murine cervical cancer cells were used for in vitro and in vivo studies, respectively. Samples included Cu-Try NPs, Cu-Try/MB NPs, SiO2 NPs, and SiO2/MB NPs for comparison. Data were acquired through UV-vis spectroscopy, fluorescence spectroscopy, confocal laser scanning microscopy, and cytotoxicity assays.

3：List of Experimental Equipment and Materials:

Equipment includes ESCALAB 250 XPS, FEI TECNAI G2 TEM, H-8100 TEM, ZEN3690 zetasizer, ThermoScientific Xseries II ICP, VARIAN CARY 50 UV-vis spectrophotometer, Leica TCS SP2 CLSM, Synergy HT microplate reader, Fluoromax-4 fluorimeter. Materials include Cu(NO3)2, NaOH, DTNB, CH3CH2OH, MB, H2DCFDA, Calcein AM, GSH, PI, DMEM, tryptone powder, and various reagents from suppliers like Beijing Chemical Reagents Company, Beyotime, Aladdin, Sigma, and Life Technologies.

4：Experimental Procedures and Operational Workflow:

Synthesis of Cu-Try NPs involved mixing tryptone with Cu(NO3)2, adding NaOH, aging at 90°C, and dialysis. Cu-Try/MB NPs were prepared by stirring MB with Cu-Try NPs and dialyzing. In vitro assays included GSH consumption detection using DTNB, ROS detection using DCFH, cytotoxicity assays with LDH kit, and cell viability tests. In vivo studies involved tumor induction in mice, intravenous injection of NPs, laser irradiation, and tumor volume measurement.

5：Data Analysis Methods:

Data were analyzed using UV-vis and fluorescence spectroscopy for absorption and emission measurements, CLSM for imaging, LDH assay for cytotoxicity, and statistical methods for in vivo tumor growth analysis.