Dataset DOI: 10.5061/dryad.pc866t23x

Description of the data and file structure

The raw data were collected from controlled in vitro and in vivo experiments designed to quantify cellular responses under defined experimental conditions. Measurements were obtained from multiple independent biological replicates and used to generate the quantitative results reported in the study. Any NA represents missing data.

Files and variables

The Dryad repository contains raw and processed datasets that are sufficient to reproduce the quantitative analyses and figures reported in this study.

Figure 1 is a schematic illustration.

This dataset contains raw data associated with Figures 2–7. Data are provided as separate Excel files for each figure. Each file contains panel-specific raw measurements, including flow-cytometry quantification, qPCR gene-expression values, fluorescence or histological image quantification, ion-release measurements, and micro-CT-derived parameters. Unless otherwise stated, values represent biological replicates, and error bars in the corresponding figures indicate mean ± SD. Statistical comparisons were performed using Tukey’s multiple-comparisons test.

figure2_raw.xlsx: Raw data used to generate Figure 2.

Description: Raw data for Figure 2, which evaluates osteo-immune responses elicited by a magnesium-releasing alginate hydrogel at a bone-regeneration site in Balb/c mice. The file includes data for time-dependent immune-cell composition within the defect, relative gene-expression heat-map values, immunofluorescence quantification of Arg1 and iNOS, TRAP staining of osteoclasts, flow-cytometry quantification of macrophage polarization, Th1/Th2 T-cell responses, and M1/M2 and Th1/Th2 ratios in control, splenectomized, and nude mice.

Variables in this file include figure panel, sample identifier, mouse model, hydrogel treatment, time point after defect creation or hydrogel injection, measured immune-cell population, measured gene or protein marker, and raw or normalized measurement value.

figure3_raw.xlsx: Raw data used to generate Figure 3.

Description: Raw data for Figure 3, which examines how strong Ca²⁺-associated TCR stimulation skews CD4⁺ T cells toward a Th1 phenotype and how extracellular Mg²⁺ reduces this bias. The file includes Nfatc1 transcript levels in the bone-defect site, IFN-γ/IL-4 ratios under different anti-CD3/CD28 bead-to-cell ratios, flow-cytometry quantification of IFN-γ⁺ and IL-4⁺ CD4⁺ T cells, Fluo-4 Ca²⁺ imaging, Mag-Fluo-4 Mg²⁺ imaging, Mg²⁺-transporter inhibition or sodium-depletion experiments, siTRPM7 and siMagt1 experiments, BTP-2 inhibition experiments, NFAT-family gene expression, NFATc1/NFATc3 and T-bet/Gata3 ratios, and cytokine-pretreatment experiments.

Variables in this file include figure panel, sample identifier, bead-to-cell stimulation ratio, extracellular Mg²⁺ concentration, inhibitor or transfection condition, cytokine pretreatment, imaging time point, measured marker, and raw or normalized value.

figure4_raw.xlsx: Raw data used to generate Figure 4.

Description: Raw data for Figure 4, which evaluates how extracellular Mg²⁺ reshapes Ca²⁺ flux signatures and effector bias of CD4⁺ T cells under moderate, low, or absent TCR stimulation. The file includes flow-cytometry quantification of IFN-γ⁺ and IL-4⁺ CD4⁺ T cells, Fluo-4 Ca²⁺ imaging parameters, NFAT-family and lineage-specifying transcription-factor expression, siTRPM7 and Mg²⁺ supplementation comparisons, SOCE-related Ca²⁺ imaging with thapsigargin, BTP-2, nifedipine, MgCl₂, STAT1/STAT6 transcript expression, and the effect of JAK inhibition on Mg²⁺-induced IFN-γ and IL-4 transcripts.

Variables in this file include figure panel, sample identifier, stimulation level, bead-to-cell ratio, extracellular Mg²⁺ concentration, inhibitor or drug treatment, transfection condition, time point, measured marker, and raw or normalized measurement value.

figure5_raw.xlsx: Raw data used to generate Figure 5.

Description: Raw data for Figure 5, which examines the effects of Mg²⁺ on osteoclast differentiation, RANKL-induced Ca²⁺ signaling, NFATc1 nuclear translocation, and macrophage M1/M2 polarization. The file includes TRAP/phalloidin-actin image quantification, osteoclast marker gene expression, Fura-2 and Fluo-4 Ca²⁺ imaging, inhibitor and siRNA perturbation experiments, NFATc1 immunofluorescence quantification, flow-cytometry analysis of macrophage polarization, iNOS and Arg1 transcript levels under different inflammatory or type-2-polarizing stimuli, and macrophage responses to conditioned CD4⁺ T-cell media.

Variables in this file include figure panel, sample identifier, cell type, extracellular Mg²⁺ concentration, differentiation or polarization stimulus, inhibitor or siRNA condition, culture time point, measured marker, and raw or normalized value.

figure6_raw.xlsx: Raw data used to generate Figure 6.

Description: Raw data for Figure 6, which evaluates the immunomodulatory capacity of magnesium-incorporated bioceramic intramedullary nails at an osteoporotic femoral fracture site. The file includes Mg²⁺ ion-release measurements from hydroxyapatite, whitlockite, and magnesium-doped whitlockite intramedullary nails, as well as day-3 and day-7 flow-cytometry quantification of Th1/Th2 T-cell responses and M1/M2 macrophage polarization in bone-marrow cells after fracture fixation.

Variables in this file include figure panel, sample identifier, mouse model, intramedullary nail type, ion-release condition, post-fracture time point, measured immune-cell population, measured marker, and raw or normalized value.

figure7_raw.xlsx: Raw data used to generate Figure 7.

Description: Raw data for Figure 7, which assesses bone regeneration and callus remodeling after osteoporotic femoral fracture fixation with hydroxyapatite, whitlockite, or magnesium-doped whitlockite intramedullary nails. The file includes four-week post-fracture micro-CT quantification, osteogenic gene-expression data, immunofluorescence quantification of IL-4 and IFN-γ in callus tissue, iNOS and Arg1 staining quantification, TRAP staining adjacent to intramedullary nail surfaces, and H&E-based histological observations of marrow surrounding each implant.

Variables in this file include figure panel, sample identifier, mouse model, intramedullary nail type, post-fracture time point, analyzed tissue region, measured marker or parameter, and raw or normalized value.

Each Excel file includes the underlying raw values used for plotting, quantification, and statistical analysis of the corresponding figure.

Code/software

Any program that will open a spreadsheet, such as Excel, is recommended.