Antioxidant Nanotherapy for Immunomodulation and Functional Remodeling of Periodontal Ligament Stem Cells for Diabetic Periodontitis in Vitro

1. Data Generation ProceduresSynthesis and Characterization of TPCDSynthesis Process:A broad-spectrum reactive oxygen species (ROS)-scavenging material, TPCD, was synthesized. The process involved multiple reaction steps, such as activation of precursors (Tpl and PBAP), chemical modification of β-cyclodextrin, precipitation, and vacuum drying.Characterization of TPCD:Fourier Transform Infrared Spectroscopy (FT-IR) was performed using a PerkinElmer FT-IR spectrometer.¹H NMR spectra were collected with a 600 MHz NMR spectrometer (Agilent).Preparation and Characterization of TPCD Nanoparticles:TPCD nanoparticles (TPCD NPs) were prepared using ethanol solutions of lecithin and DSPE-PEG, mixed with an aqueous solution containing TPCD. Particle size and surface morphology were determined with:Malvern Zeta potential analyzer for particle size (Nano ZS90, Malvern).Transmission electron microscopy (TEM, TECNAI-10, Philips).Scanning electron microscopy (SEM, FIB-SEM, Zeiss Crossbeam 340).In Vitro ROS-Scavenging ActivitiesGradient concentrations of TPCD NPs were tested for free-radical scavenging capabilities (DPPH•, superoxide anion, hydrogen peroxide, and hydroxyl radicals) using respective commercial detection kits. Absorbance values were measured using an Infinite M200 Pro plate reader (TECAN).Isolation and Culture of Periodontal Ligament Stem Cells (hPDLSCs):Periodontal ligament stem cells (hPDLSCs) were isolated from teeth under sterile conditions. Three groups were defined: (1) Healthy hPDLSCs (P), (2) hPDLSCs from periodontitis patients (PP), (3) hPDLSCs from patients with diabetes mellitus and periodontitis (DPP).A summary of criteria (e.g., patient age, inclusion, and exclusion criteria) is documented.2. Data Processing Methods and StepsData Capture:Primary data for characterizations, such as FT-IR, NMR spectra, TEM/SEM images, and particle-size distributions, were collected using the respective instruments' software.Assay-based quantified data (e.g., ROS detection, ALP staining, and cell viability) were collected using spectrophotometers, fluorescence microscopes, and image analysis software.Data Cleaning and Transformation:Raw data were cleaned by removing experimental artifacts and normalizing values with baseline corrections.Image files from ALP staining, Alizarin Red staining, and TEM/SEM were analyzed using ImageJ for semi-quantitative measurements.3. Devices and Tools UsedHardware and Instruments:PerkinElmer FT-IR spectrometer for FT-IR.Agilent DD2 600 MHz NMR spectrometer for ¹H NMR.Malvern Nano ZS90 Zeta potential analyzer for particle size analysis.Philips TECNAI-10 TEM and Zeiss FIB-SEM for microscopy imaging.Tecan Infinite M200 Pro plate reader for absorbance measurements.BD Biosciences FACS Calibur for flow cytometry.Software Tools:ImageJ (for staining analyses).R (packages: limma, clusterProfiler, pheatmap, ggplot2) for statistical and bioinformatics analysis.SPSS 26.0 for statistical significance testing.4. Temporal and Geographical ScopesGeographical Scope:Teeth were collected from patients at Zunyi Medical University Hospital, Zunyi, China.Temporal Scope:Sample collection and corresponding analyses were conducted between [start date] and [end date].Temporal Resolution:Time points for analyses spanned days 1, 7, 14, and 21 for gene/protein expression and differentiation experiments.5. Tabular DataTotal Number of Entries:Entry numbers vary by dataset based on experimental analysis, e.g., levels of ROS scavenged, ALP/Alizarin Red staining intensities, RNA expression levels, and PCA data.