A logic-gated nanoplatform spatiotemporally orchestrates immunometabolic reprogramming and osteogenesis for diabetic periodontal regeneration

Abstract

Achieving periodontal regeneration in diabetic periodontitis (DPD) is hindered by a self-perpetuating vicious cycle of excessive oxidative stress and adaptive immune dysfunction. Here, using clinical samples, multi-omics profiling, and DPD rat model, we identified an association between oxidative stress and CD4⁺ T cell dysregulation. This process was characterized by reactive oxygen species (ROS) accumulation and Th17/Treg imbalance, which may contribute to progressive alveolar bone loss. To modulate this pathological process, we engineered a logic-gated hydrogel-nanoparticle hybrid system for hierarchical delivery of active vitamin D (AVD). This platform features a dynamic boronic ester crosslinked network that functions as a ROS-responsive switch. Upon exposure to the oxidative microenvironment, the matrix undergoes programmed degradation to scavenge ROS, triggering the rapid release of antibody-functionalized nanoparticles. These nanoparticles selectively target CD4⁺ T cells and deliver AVD intracellularly, where it modulates mitochondrial homeostasis and metabolic signaling pathways, thereby restoring T-cell immune homeostasis. Following this immune reset, the system transitions into a second phase where porous copper-based metal-organic frameworks provide sustained release of AVD and osteogenic Cu²⁺ to support long-term bone remodeling. By integrating redox-responsive actuation, targeted immunometabolic regulation, and sustained osteoimmunomodulation, this hierarchical system provides a promising therapeutic strategy for periodontal regeneration in diabetic periodontitis.

Graphical abstract

Highlights

• •Oxidative stress-driven CD4⁺ T cell dysfunction accelerates DPD bone loss.
• •A ROS-responsive hydrogel enables hierarchical AVD delivery in DPD.
• •CD4-targeted AVD delivery restores T cell metabolism and immune balance.
• •pCu-MOF depots provide sustained AVD/Cu²⁺ release for bone remodeling.
• •AVD@pCuMOF/BSA-AP promotes diabetic periodontal regeneration in vivo.

1. Introduction

The precise spatiotemporal orchestration of immune and regenerative niches remains a grand challenge in nanomedicine, particularly for chronic inflammatory diseases complicated by metabolic disorders such as type 2 diabetes (T2D) [[1], [2], [3]]. In such pathological environments, tissue regeneration is often arrested by a self-perpetuating vicious cycle involving oxidative stress, immune dysregulation, and impaired stem cell function [4,5]. Diabetic periodontitis (DPD), a hallmark complication of T2D characterized by accelerated alveolar bone loss, serves as a paradigmatic model of this therapeutic dilemma. While conventional strategies focus on broad-spectrum anti-inflammatory or osteogenic agents [[5], [6], [7]], they frequently fail to restore homeostasis. This failure stems from an inability to dismantle the complex, multi-layered pathogenic network wherein hyperglycemia-induced reactive oxygen species (ROS) are associated with metabolic dysfunction and pathogenic polarization (Th17/Th1) of CD4⁺ T cells [[8], [9], [10], [11], [12]], which in turn amplifies inflammation and fuels RANKL-mediated bone resorption [[13], [14], [15], [16]]. Thus, effective regeneration necessitates a hierarchical intervention strategy involving a rapid, targeted reset of the immune metabolic state to break the inflammatory cycle, followed by a sustained, permissive rebuilding of the osteogenic microenvironment.

Achieving this level of control requires advanced nanotherapeutic platforms capable of executing logic-gated responses within the physiological milieu. Current delivery systems, however, often lack the intelligence to distinguish between disease phases or to deliver distinct therapeutics with stage-specific kinetics [[17], [18], [19], [20], [21], [22], [23]]. Active vitamin D (AVD) and its analogs (e.g., Eldecalcitol) have emerged as potent immunometabolic modulators capable of restoring Th17/Treg balance and promoting osteogenesis [[24], [25], [26], [27], [28]]. However, their clinical application is severely limited by short half-life, poor cellular uptake, and the risk of systemic toxicity (e.g., hypercalcemia) from burst release [29]. To harness the full potential of AVD, it must be delivered intracellularly to specific immune subsets during the early inflammatory phase, while maintaining a sustained extracellular presence during the later remodeling phase. This spatiotemporal requirement presents a unique opportunity for nanotechnology-enabled hierarchical delivery. Injectable hydrogels are particularly appealing due to their extracellular matrix-mimicking architecture and ability to conform to irregular periodontal pockets [30]. Notably, stimuli-responsive hydrogels that adapt to dynamic microenvironmental cues represent an advanced platform for on-demand, precise therapeutic delivery [31,32].

Here, we report a logic-gated hydrogel-nanoparticle hybrid composite system designed to modulate the osteoimmune microenvironment in DPD (Fig. 1). We first identify an oxidative stress-associated CD4⁺ T cell dysregulation cycle as a critical and druggable target through clinical sample analysis, multi-omics profiling, and animal models. To target this mechanism, we constructed a microenvironment-adaptive platform (AVD@pCuMOF/BSA-AP) that integrates two distinct therapeutic nanomodulators within a ROS-responsive hydrogel network. Distinct from conventional single-stimulus responsive systems, this platform operates through a two-input logic framework. The first input is the elevated ROS level characteristic of the DPD microenvironment, which serves as a licensing signal to trigger degradation of the dynamic boronic ester-crosslinked hydrogel network and initiate release of the antibody-functionalized nanoparticles. The second input is the presence of CD4⁺ T lymphocytes expressing the specific surface epitopes recognized by the antibody-functionalized nanoparticles, which enhances selective cellular internalization and intracellular drug delivery. The therapeutic cascade is maximally activated and enhanced when these two conditions occur in sequence. This logic-controlled activation simultaneously interrupts the oxidative stress–T cell dysfunction loop and restores immune homeostasis through mitochondrial and metabolic reprogramming. Meanwhile, exposed polyphenol-engineered MOFs act as ROS scavengers and sustained reservoirs for AVD and osteogenic Cu²⁺, thereby coupling early immunomodulation with long-term bone regeneration. By integrating molecular recognition, dynamic chemistry, and immunometabolic regulation, this study presents a promising therapeutic strategy for periodontal regeneration in diabetic periodontitis.

Fig. 1.

Designing a ROS-responsive hydrogel nanoplatform (AVD@pCuMOF/BSA-AP) for immunomodulation and periodontal regeneration in diabetic periodontitis.

a Schematic illustration of the synthesis of the AVD@pCuMOF/BSA-AP hydrogel, and its minimally invasive administration into the periodontal pocket via local injection.

b The reactive oxygen species (ROS)-responsive AVD@pCuMOF/BSA-AP hydrogel enables sequential and hierarchical active vitamin D (AVD) delivery, precisely targeting CD4⁺ T cells and maintaining sustained long-term release. By scavenging ROS, modulating CD4⁺ T cell immune homeostasis, and supporting osteogenesis while attenuating osteoclastogenesis, this platform improves the oxidative stress-associated immune imbalance and supports alveolar bone repair in diabetic periodontitis (DPD).

2. Results and discussion

2.1. Oxidative stress-associated CD4⁺ T cell dysregulation is linked to alveolar bone loss in DPD

To elucidate the impact of the oxidative stress in DPD, we first analyzed ROS accumulation in gingival tissues collected from DPD patients using dihydroethidium (DHE) staining (Fig. 2a). Compared to healthy control tissues, the patient-derived gingival specimens exhibited significantly elevated ROS levels (Fig. 2b; Fig. S1), which was further validated by detecting substantially higher hydrogen peroxide (H₂O₂) concentration in DPD gingival tissue lysates (Fig. 2c). These findings indicate the presence of a hyper-oxidative microenvironment in DPD-affected gingival tissues. These findings are consistent with previous reports indicating that hyperglycemia-associated metabolic disturbances and inflammatory conditions may contribute to redox imbalance and immune dysregulation during diabetic periodontitis [33,34]. To characterize diabetes-associated alterations in the gingival immune landscape, we analyzed a publicly available gingival scRNA-seq dataset from db/db mice (GEO: GSE188217), which were not subjected to experimental periodontitis induction. (Fig. 2d). Uniform manifold approximation and projection (UMAP) visualization of 18,743 cells resolved major leukocyte populations (Fig. 2e), with subsequent sub-clustering of the T cell compartment identifying diverse lineages, including CD4⁺, CD8⁺, γδT (GDT), NKT and ILC2 subsets (Fig. 2f; Fig. S2). Notably, CD4⁺ T cells showed an increased proportion in diabetic mice (24.35% versus 16.42% in controls; Fig. 2g), representing a prominent altered immune population under diabetic conditions. This expansion was accompanied by the activation of IL-17, TNF, NF-κB, and JAK-STAT signaling pathways, alongside the upregulation of T cell exhaustion (PD-L1/PD-1) and differentiation markers (Fig. 2h). Complementary bulk RNA sequencing of splenic CD4⁺ T cells from db/db mice further characterized diabetes-associated intrinsic transcriptional alterations of CD4⁺ T cells (Fig. 2i; Fig. S3). Differential expression analysis highlighted a coordinated upregulation of pro-inflammatory effectors (Il6, Il22, Ifng, Csf2), Th17-polarizing factors (Rorc, Runx1) and exhaustion markers (Tcf7, Lef1, Havcr2, Ctla4, Pdcd1), contrasted with suppression of Treg-associated mediators (Tgfb1, Foxp3, Il2ra, Stat5; Fig. 2j). The integration of single-cell transcriptomic profiling and CD4⁺ T cell transcriptomic analysis enabled characterization of both immune landscape alterations and CD4⁺ T cell intrinsic functional changes under diabetic conditions. Together, these multi-omics data revealed alterations in CD4⁺ T cell states characterized by pro-inflammatory and exhaustion-associated phenotypes with reduced regulatory capacity, which may contribute to chronic inflammation and tissue destruction in DPD [27,35].

Fig. 2.

Heightened oxidative stress, CD4⁺ T cell dysfunction, and alveolar bone loss in diabetic periodontitis.

a Schematic workflow of human gingival tissues collection and subsequent analyses.

b Representative immunofluorescence images of DHE staining (red) in gingival tissues from healthy donors and patients with diabetic periodontitis. Nuclei were stained with DAPI (blue). DHE, dihydroethidium; DAPI, 4′,6-diamidino-2-phenylindole. Scale bar, 50 μm.

c Quantification of hydrogen peroxide (H₂O₂) concentrations in gingival tissues from healthy individuals and patients with diabetic periodontitis.

d Schematic of single-cell sequencing (GSE188217) of gingival tissues from control (Ctrl) and type 2 diabetes (T2D) mice.

e, f Uniform manifold approximation and projection (UMAP) visualization showing major cell clusters (e) and re-clustered T cell subsets (f).

g Relative cell proportions of T cell subsets. Black dotted boxes indicate the proportion of CD4⁺ T cell fraction.

h KEGG enrichment analysis of DEGs in gingival CD4⁺ T cells. KEGG, Kyoto Encyclopedia of Genes and Genomes; DEGs, differentially expressed genes.

i Schematic workflow for CD4⁺ T cell isolation from spleens of Ctrl and T2D mice followed by bulk RNA-seq analysis.

j Relative expression of DEGs related to cytokines, T cell differentiation, and T cell exhaustion (Tex) in splenic CD4⁺ T cells.

k Representative 3D micro-CT images and buccal-palatal sectional views of the maxillary alveolar bone in Ctrl and DPD rats. Scale bar, 2 mm 

l Representative DHE staining of gingival tissues from Ctrl and DPD rats. Scale bar, 50 μm 

m Flow cytometric analysis of Th17 (IL-17A⁺), Th1 (IFN-γ⁺), and exhausted T cells (CTLA-4⁺) CD4⁺ T cells in gingival tissues from Ctrl and DPD rats. IL-17A, interleukin-17; IFN-γ, interferon-gamma; CTLA-4, cytotoxic T-lymphocyte-associated protein 4.

n Pearson correlation analysis between cementoenamel junction to alveolar bone crest (CEJ-ABC) distance and T cell subset frequencies.

Data are presented as mean ± standard deviation. n = 3 human donors (b-c), n = 3 mice per group (i), and n = 12 mice for correlation analysis (n). Statistical significance was determined using unpaired two-tailed Student's t-test. Correlation analysis was performed using Pearson correlation. *P < 0.05 was considered statistically significant. Schematics were created using BioRender.com.

We validated these findings in a DPD rat model, where micro-CT analysis confirmed severe alveolar bone loss (Fig. 2k), coincident with DHE-detectable oxidative stress accumulation in periodontal tissues (Fig. 2l). Immunophenotyping revealed a sharp expansion of pathogenic subsets (IL-17A⁺: 3.1-fold, IFN-γ⁺: 6.5-fold, P < 0.001) and a depletion of regulatory populations (forkhead box P3, FOXP3⁺: 70.5% reduction, P < 0.001; Fig. 2m; Fig. S4). Notably, these immune shifts correlated strongly with bone destruction metrics (r = 0.83-0.92, P < 0.001; Fig. 2n; Fig. S5), establishing quantitative links between immune dysregulation and osteolytic progression [28,36]. Taken together, our findings suggest that oxidative stress-associated immune dysregulation represents an important feature of diabetic periodontitis, with CD4⁺ T cell imbalance characterized by Th17/Th1 polarization, exhaustion-associated alterations, and impaired regulatory signatures. These observations support the development of therapeutic strategies targeting both redox imbalance and CD4⁺ T cell immune homeostasis. While our findings establish a robust association between redox dysregulation and CD4⁺ T -cell dysfunction, causal relationships remain to be mechanistically resolved. Future studies employing targeted oxidative modulation and immune rescue strategies will be essential to delineate the directionality and hierarchy of this interaction.

2.2. Engineering the hierarchical hydrogel-nanoparticle hybrid

To realize the programmed spatiotemporal delivery, we engineered two distinct nanomodulators with tailored physicochemical properties and release kinetics. First, we constructed CD4-targeting and immune-resetting nanoparticles (AVD@BSA-mAb NPs) via a desolvation-crosslinking strategy (Fig. 3a). These AVD@BSA-mAb NPs exhibited a uniform diameter of ∼256 nm, were surface-functionalized with anti-CD4 antibodies via thiol-maleimide chemistry, as confirmed by a distinct zeta potential inversion (−48.1 to −7.5 mV) and SDS-PAGE analysis (Fig. 3b–e; Fig. S6). The hydrodynamic diameter and Zeta potential of BSA NPs remained stable over the 3-day incubation period in both PBS and artificial saliva, confirming the nanoparticles possess favorable colloidal stability under simulated physiological and pathological conditions (Fig. S7). Crucially, the nanoscale size and specific surface functionalization promoted CD4⁺ T cell uptake in vitro and enhanced nanoparticle accumulation within gingival CD4⁺ T cells in vivo following periodontal administration (Fig. 3f; Fig. S8), while maintaining excellent biocompatibility at concentrations ≤ 100 μg/mL (Fig. 3g; Fig. S9), ensuring precise intracellular delivery of the immunometabolic modulator AVD.

Fig. 3.

Synthesis and characterization of AVD@BSA-mAb NPs, AVD@pCu-MOFs and AVD@pCuMOF/BSA-AP hydrogel.

a Schematic illustration showing the synthesis of AVD-loaded monoclonal antibody-conjugated bovine serum albumin nanoparticles (AVD@BSA-mAb NPs).

b TEM image of BSA NPs. Scale bar, 500 nm.

c Zeta potential measurements of BSA NPs and BSA-mAb NPs.

d Encapsulation efficiency and loading efficiency of AVD within BSA-mAb NPs.

e SDS-PAGE analysis with Coomassie brilliant blue staining confirming antibody conjugation. Red arrows, BSA; black arrows, CD4 mAb heavy chain; white arrows, CD4 mAb light chain.

f Representative fluorescence microscopy images showing enhanced association of BSA-mAb NPs with CD4⁺ T cells. Nuclei were stained with DAPI (blue), and nanoparticles (BSA NPs and BSA-mAb NPs) were labeled with Rhodamine B (RhB, red). Scale bar, 5 μm 

g Live/dead staining images of CD4⁺ T cells after 48 h of co-culture with AVD@BSA-mAb NPs. Scale bar, 200 μm 

h Schematic illustration showing the synthesis of AVD-loaded polyphenolic copper-based metal-organic frameworks (AVD@pCu-MOFs).

i TEM image of pCu-MOFs. Scale bar, 100 nm.

j Encapsulation efficiency and loading efficiency of AVD within pCu-MOFs.

k, l XPS survey (k) and high-resolution Cu 2p spectra (l).

m Antioxidant activity of pCu-MOFs assessed by DPPH, ·OH and ·O₂ scavenging assays at concentrations of 100, 200, 300 μg/mL.

n Oxygen (O₂) generation kinetics of pCu-MOFs in the presence of 10 mM H₂O₂.

o Schematic illustration of the fabrication of AVD@pCuMOF/BSA-AP hydrogel.

p EDS elemental mapping of pCuMOF/BSA-AP hydrogel. Scale bar, 1 μm.

q Rheological analysis showing the shear-thinning behavior of the pCuMOF/BSA-AP hydrogel (viscosity vs. shear rate from 1 to 50 s⁻¹) at 25°C.

r H₂O₂-responsive degradation profiles of pCuMOF/BSA-AP hydrogel.

s, t Release kinetics of pCuMOF/BSA-AP hydrogel under different stimuli by using VD3 (s) and RhB (t) as model drugs.

u Biocompatibility assessment via live/dead staining of CD4⁺ T cells and human gingival fibroblasts (HGFs) after 48 h. Scale bar, 25 μm or 100 μm.

Data are presented as mean ± standard deviation. All experiments were independently repeated three times (n = 3). Schematics created with BioRender.com.

Parallelly, we synthesized the osteo-inductive metal-organic frameworks (pCu-MOFs) via solvothermal self-assembly, stabilized by a poly (tannic acid) (PTA) coating (Fig. 3h and i; Fig. S10–S12). The PTA coating not only enhanced colloidal stability but also introduced a rich density of phenolic hydroxyls, conferring potent intrinsic ROS-scavenging activity (Fig. 3m and n; Fig. S13). These pCu-MOFs exhibited a high loading capacity for AVD (1.25%) and mixed Cu(I)/Cu(II) valence states (Fig. 3j–l), serving as a robust reservoir for long-term therapeutic release.

These two functional nanomodules were integrated into a dynamic hydrogel matrix composed of phenylboronic acid-grafted alginate (ALG-PBA) and gallic acid-grafted polylysine (PLL-GA; Fig. 3o). Specifically, we first synthesized ALG-PBA and PLL-GA. The as-synthesized ALG-PBA and PLL-GA were characterized by Fourier transform infrared (FTIR) and ¹H NMR spectroscopy (Fig. S14). SEM imaging confirmed the uniform incorporation of spherical nanoparticles within the matrix (Fig. S15), while elemental mapping of Cu and S validated their distribution (Fig. 3p). Rheological and mechanical characterization confirmed that the hybrid AVD@pCuMOF/BSA-AP hydrogel possesses shear-thinning injectability (Fig. 3q), rapid self-healing, and tissue adhesiveness (Fig. S16), allowing seamless adaptation to irregular periodontal defects. The pCuMOF/BSA-AP hydrogel maintains structural integrity and injectability across the physiological oral pH range and at body temperature, supporting its suitability for periodontal therapy (Fig. S17).

The boronic ester bonds served as the ROS-logic gate under oxidative conditions (e.g., DPD microenvironment). Under these conditions, these bonds underwent triggered cleavage, leading to accelerated matrix degradation (Fig. 3r) and broad-spectrum ROS scavenging (Fig. S18). This ROS-triggered degradation facilitated a programmed drug release profile, where the model cargo Rhodamine B (RhB) from BSA NPs was released rapidly (47.67 ± 4.55% in H₂O₂ vs. 40.79 ± 6.22% in PBS), whereas MOF-encapsulated cargo exhibited a strictly controlled, sustained release (11.18 ± 2.30% in H₂O₂ vs. 8.41 ± 1.87% in PBS; Fig. 3s and t). This sequential release capability allows the hydrogel to address the temporal requirements of periodontal regeneration by facilitating immediate immune reprogramming followed by long-term osteo-metabolic regulation.

Finally, the biocompatibility of the AVD@pCuMOF/BSA-AP hydrogel was evaluated on four critical cell types implicated in periodontal homeostasis and immune regulation, including CD4⁺ T cells, human gingival fibroblasts (HGFs), periodontal ligament stem cells (PDLSCs), and RAW264.7 macrophages. Both Live/dead staining and CCK-8 viability assays confirmed the safety of the AVD@pCuMOF/BSA-AP hydrogel with cell viability above 95% after 48 h of co-culture with (Fig. 3u; Fig. S19), validating its potential for clinical translation.

2.3. AVD@pCuMOF/BSA-AP hydrogel attenuates oxidative inflammatory stress in DPD microenvironment

Because the diabetic periodontal microenvironment is characterized by oxidative stress and inflammatory activation [33], we first evaluated whether AVD@pCuMOF/BSA-AP hydrogel could attenuate this pathological microenvironment in supporting in vitro models. HGFs and macrophage cells were stimulated with high glucose plus lipopolysaccharide (HG + LPS) to mimic diabetic inflammatory conditions (Fig. 4a). Flow cytometry analysis with the DCFH-DA probe revealed superior ROS scavenging capability of AVD@pCuMOF/BSA-AP hydrogel, which exhibited the lowest fluorescence intensity among all test groups (Fig. 4b and c). This potent antioxidant effect was mirrored in RAW264.7 macrophages (Fig. S20), indicating superior ROS scavenging ability of AVD@pCuMOF/BSA-AP hydrogel, attributed to combinatorial antioxidant actions of its components.

Fig. 4.

In vitro antioxidative and anti-inflammatory effects of AVD@pCuMOF/BSA-AP hydrogel.

a Schematic of the transwell co-culture system used to evaluate the antioxidant activity of AVD@pCuMOF/BSA-AP hydrogel in HGFs.

b, c Flow cytometric analysis (b) and quantification (c) of intracellular ROS levels (DCFH-DA) in HGFs.

d Representative immunofluorescence images of mitochondrial membrane potential assessed by JC-1 staining (Red: aggregates/healthy; Green: monomers/damaged). Nuclei were stained with Hoechst (blue) in HGFs. Scale bar, 20 μm.

e Quantification of the JC-1 aggregate-to-monomer fluorescence ratio.

f Representative confocal images showing mitochondrial mass (Mito-Tracker, green) and mitochondrial superoxide (MitoSOX, red) in HGFs. Nuclei were stained with Hoechst (blue).

g Quantification of MitoSOX mean fluorescence intensity (MFI) in HGFs. Scale bar, 10 μm 

h Heatmap showing relative mRNA expression levels of antioxidant enzymes (SOD2, CAT, and GPX1) in HGFs. SOD2, superoxide dismutase 2; CAT, catalase; GPX1, glutathione peroxidase 1.

i Western blot analysis of SOD2 and CAT protein expression in HGFs.

j Schematic illustration for the anti-inflammatory effect of AVD@pCuMOF/BSA-AP.

k ELISA quantification inflammatory cytokines (IL-1β, IL-6) in HGF culture supernatants. IL-1β, interleukin-1 beta; IL-6, interleukin-6.

l Heatmap of pro-inflammatory gene expression levels (IL-1β, IL-6 and TNF-α) in HGFs. TNF-α, tumor necrosis factor-alpha.

m Western blot analysis of IL-6 and IL-1β protein expression in HGFs.

n Representative immunofluorescence images of TLR4 expression (green) in HGFs. Nuclei: DAPI (blue). TLR4, toll-like receptor 4. Scale bar, 50 μm.

Experimental groups were defined as follows: I, Ctrl; II, HG + LPS; III, AVD; IV, AP hydrogel; V, pCuMOF/BSA-AP hydrogel; VI, AVD@pCuMOF/BSA-AP hydrogel. Data are presented as mean ± standard deviation. n = 3 independent biological replicates per group. Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparison test. *P < 0.05 was considered statistically significant. Schematics were created using BioRender.com.

Mitochondrial dysfunction, a hallmark of DPD, was further assessed using JC-1 staining, where depolarized mitochondria exhibit decreased red/green fluorescence ratios, indicating membrane potential loss [37]. Strikingly, treatment with AP, pCuMOF/BSA-AP, and AVD@pCuMOF/BSA-AP restored mitochondrial integrity, with the composite AVD@pCuMOF/BSA-AP hydrogel displaying the highest preservation of ΔΨm (Fig. 4d and e; Fig. S21). Mito-Tracker/MitoSOX double staining further corroborated these findings, showing that the DPD conditions induced mitochondrial fragmentation and elevated mitochondrial ROS (mtROS) levels. In contrast, the AVD@pCuMOF/BSA-AP hydrogel restored mitochondrial morphology and significantly reduced mtROS overload (Fig. 4f and g; Fig. S22). Mechanistically, this protection was linked to the reactivation of endogenous antioxidant defenses [38]. As shown in Fig. 4h,i and Fig. S23, qRT-PCR and Western blot analyses confirmed a robust upregulation of SOD2, CAT, and GPX1 at both the mRNA and protein levels, suggesting that the AVD@pCuMOF/BSA-AP hydrogel not only scavenges exogenous ROS but also potentiates cellular resilience.

In parallel with its antioxidant effects, the AVD@pCuMOF/BSA-AP hydrogel also displayed robust anti-inflammatory ability in HG + LPS-stimulated HGFs as models (Fig. 4j). Quantitative ELISA analysis showed that the AVD@pCuMOF/BSA-AP hydrogel significantly downregulated the secretion of IL-1β and IL-6 (Fig. 4k), outperforming individual component treatments (AVD, AP, or pCuMOF/BSA-AP alone). These observations were further supported by qRT-PCR and Western blot analyses (Fig. 4l and m; Fig. S24). Importantly, immunofluorescence staining revealed that AVD@pCuMOF/BSA-AP hydrogel effectively suppressed HG + LPS-induced toll-like receptor 4 (TLR4) overexpression (Fig. 4n; Fig. S25). Since TLR4 activation in GFs triggers the release of proinflammatory cytokines and proteolytic enzymes that accelerate tissue breakdown [39], this suppression highlights the potential of AVD@pCuMOF/BSA-AP hydrogel to interrupt the upstream inflammatory cascade driving DPD progression. These findings indicated that AVD@pCuMOF/BSA-AP hydrogel attenuated oxidative and inflammatory stress in periodontal microenvironment-related cells, providing a favorable context for subsequent CD4⁺ T cell immune regulation.

2.4. AVD@pCuMOF/BSA-AP hydrogel reshapes CD4⁺ T cell immune homeostasis through metabolic pathway reprogramming

CD4⁺ T cell dysregulation, characterized by Th17 polarization, impaired Treg balance, and exhaustion-associated phenotypes, emerged as a central immune feature of DPD in our omics and in vivo analyses, consistent with previous evidence implicating CD4⁺ T cell imbalance in diabetic periodontitis progression [27,40]. Therefore, we next focused on CD4⁺ T cells as the primary immunoregulatory target of AVD@pCuMOF/BSA-AP hydrogel. To evaluate its immunomodulatory effects, we performed transcriptomic and functional profiling of murine CD4⁺ T cells under simulated diabetic inflammatory conditions (Fig. 5a; Fig. S26). RNA-seq analysis revealed a distinct gene expression profile in the AVD@pCuMOF/BSA-AP (VI) group compared with the HG + LPS (II) group (Fig. 5b). Gene Ontology (GO) enrichment analysis indicated that the AVD@pCuMOF/BSA-AP treatment upregulated critical immunoregulatory pathways including IL-17 suppression, Th17 cell differentiation inhibition, mitochondrial homeostasis, and Treg cell development, while simultaneously downregulating pathways involved in T cell dysfunction and oxidative stress responses (Fig. 5c). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed the multifaceted immunometabolic effect of AVD@pCuMOF/BSA-AP hydrogel, identifying significant modulation of IL-17 signaling, T cell receptor signaling, and key metabolic pathways such as fatty acid degradation and mitophagy (Fig. 5d). These findings suggest that the AVD@pCuMOF/BSA-AP hydrogel may act by recalibrating the mitochondrial metabolic machinery to support immune homeostasis.

Fig. 5.

Transcriptomic and metabolic reprogramming of CD4⁺ T cells by AVD@pCuMOF/BSA-AP hydrogel.

a Schematic workflow of CD4⁺ T cell treatment and analysis.

b Volcano plot of DEGs (AVD@pCuMOF/BSA-AP hydrogel treatment vs. HG + LPS).

c, d Functional enrichment analysis via GO (c) and KEGG (d). GO, Gene Ontology.

e Flow cytometric analysis of CD4⁺ FOXP3⁺ regulatory T cells (Tregs) and CD4⁺ IL-17A⁺ Th17 cells.

f, g Flow cytometric analysis of T cell exhaustion markers CTLA-4 (f) and PD-1 (g) in CD4⁺ T cells. PD-1, programmed cell death protein 1.

h Quantification of the frequencies of FOXP3⁺, IL-17A⁺, CTLA-4⁺ and PD-1⁺ CD4⁺ T cells. FOXP3, forkhead box P3.

i Representative immunofluorescence images of CD4⁺ T cells for mitochondrial for mitochondrial mass (Mito-Tracker Green) and the fatty acid oxidation enzyme CPT1A (red). Nuclei were stained with Hoechst/DAPI (blue). Scale bar, 5 μm.

j Quantification of MFI for Mito-Tracker and CPT1A.

k Schematic illustration showing that AVD@pCuMOF/BSA-AP hydrogel restores CD4⁺ T cell immune homeostasis, accompanied by reduced oxidative stress and metabolic remodeling.

Experimental groups were defined as follows: I, Ctrl; II, HG + LPS; III, AVD; IV, AP hydrogel; V, pCuMOF/BSA-AP hydrogel; VI, AVD@pCuMOF/BSA-AP hydrogel. Data are presented as mean ± standard deviation. n = 3 independent biological replicates per group. Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparison test. *P < 0.05 was considered statistically significant. Schematics were created using BioRender.com.

Functional assays corroborated these transcriptomic findings, demonstrating that the AVD@pCuMOF/BSA-AP hydrogel attenuated DPD-associated immune pathology. Flow cytometry analysis demonstrated a marked suppression of pro-inflammatory IL-17A expression (17.8% reduction) and T cell exhaustion markers (CTLA-4: 65.5% reduction; PD-1: 31.8% reduction), accompanied by a significant expansion of FOXP3+ Tregs (1.2-fold increase) (Fig. 5e–h; Fig. S27). These findings illustrate AVD@pCuMOF/BSA-AP hydrogel possesses dual immunomodulatory capacity, combining the intrinsic immunoregulatory properties of AVD with the antioxidant components of AP hydrogel [41,42], which effectively counteracts chronic antigen-driven CD4⁺ T cell dysfunction [43].

Metabolic analyses suggested that AVD@pCuMOF/BSA-AP hydrogel treatment was associated with alterations in mitochondrial quality control and fatty acid metabolism, including mitophagy-related changes (67.5% reduction in Mito-Tracker signal) and carnitine palmitoyltransferase 1A (CPT1A)-associated fatty acid oxidation (FAO, approximately 16.3-fold upregulation; Fig. 5i and j). Together with the transcriptomic enrichment of mitophagy- and FAO-related pathways, these findings suggest that hydrogel-mediated restoration of CD4⁺ T cell immune homeostasis is associated with metabolic remodeling of mitochondrial function and lipid utilization. Given the known importance of mitochondrial quality control and FAO in T cell fate decisions [44,45], these metabolic changes may contribute to the observed increase in FOXP3⁺ Tregs and reduction in IL-17A⁺ Th17 cells. By modulating metabolism-associated pathways in CD4⁺ T cells, the AVD@pCuMOF/BSA-AP hydrogel contributes to immune homeostasis and creates a more favorable microenvironment for inflammation control and tissue repair (Fig. 5k). However, as direct causal validation using mitophagy or FAO inhibition was not performed in the present study, mitophagy- and FAO-related changes are interpreted as associated metabolic features rather than upstream drivers of Treg/Th17 rebalancing.

2.5. AVD@pCuMOF/BSA-AP hydrogel promotes bone remodeling through CD4⁺ T cell-mediated osteoimmunomodulation

To investigate whether CD4⁺ T cell mediated immune modulation is associated with improved bone remodeling, we further assessed the effects of AVD@pCuMOF/BSA-AP hydrogel on osteogenic and osteoclastic responses using PDLSCs and bone marrow-derived macrophages (BMDMs) as downstream functional effector models. In the context of DPD, oxidative stress propagates inflammation to the alveolar bone, fueling a destructive imbalance between osteoblast suppression and osteoclast activation [6]. We first assessed the direct osteogenic impact of the AVD@pCuMOF/BSA-AP hydrogel on human PDLSCs due to their superior osteogenic/chondrogenic differentiation potential compared to other mesenchymal stem cells (Fig. 6a) [46]. Following 7 days of osteogenic induction, PDLSCs treated with the AVD@pCuMOF/BSA-AP hydrogel exhibited significantly higher alkaline phosphatase (ALP) activity compared to those treated with individual formulations (AP, pCuMOF/BSA-AP) or AVD-alone (Fig. 6b–d). By day 21 the hydrogel promoted robust matrix mineralization, evidenced by extensive Alizarin Red S (ARS)-positive calcium deposition (Fig. 6c–e). qRT-PCR analysis further demonstrated that AVD@pCuMOF/BSA-AP treatment orchestrated the coordinated upregulation of key osteogenic marker across differentiation stages (Fig. 6f), including ALP (an early differentiation), runt-related transcription factor 2 (RUNX2, master transcriptional regulator), osteopontin (OPN, late-stage mineralization), and type I collagen (COL1, matrix component). This transcriptional activation was subsequently validated at the protein level via Western blot analyses (Fig. 6g; Fig. S28) and confirmed by immunofluorescence visualization of OPN distribution (Fig. S29).

Fig. 6.

AVD@pCuMOF/BSA-AP hydrogel regulates osteogenic-osteoclastic balance in vitro.

a Schematic workflow of periodontal ligament stem cells (PDLSCs) and bone marrow-derived macrophages (BMDMs) isolation and subsequent osteogenic or osteoclastogenic induction.

b Representative alkaline phosphatase (ALP) staining images in PDLSCs after 7 days of osteogenic induction. Scale bar, 100 μm.

c Representative Alizarin Red S (ARS) staining images of mineralized nodules in PDLSCs after 21 days of osteogenic induction. Scale bar, 100 μm 

d Quantification of ALP activity in PDLSCs at day 7.

e Quantification of calcium deposition based on ARS staining at day 21.

f Heatmap of mRNA expression levels of osteogenic markers (ALP, RUNX2, OPN and COL1) in PDLSCs. RUNX2, runt-related transcription factor 2; OPN, osteopontin; COL1, type I collagen.

g Western blot analysis of osteogenic proteins (ALP, RUNX2, OPN) in PDLSCs.

h Representative TRAP staining images showing multinucleated osteoclasts derived from RANKL-induced BMDMs. TRAP, tartrate-resistant acid phosphatase; RANKL, receptor activator of nuclear factor-kappa B ligand. Scale bar, 100 μm.

i Quantification of osteoclast area (TRAP-positive area percentage).

j Representative immunofluorescence staining of F-actin rings (red; DAPI, blue) in BMDMs-derived multinucleated osteoclasts. Scale bar, 100 μm 

k Heatmap showing mRNA expression levels of osteoclastogenic markers (MMP9 and CTSK) in BMDMs. MMP9, matrix metalloproteinase-9; CTSK, cathepsin K.

l Western blot analysis of MMP9 and CTSK protein expression in BMDMs.

m Schematic of the transwell co-culture system to evaluate the indirect regulation of PDLSCs and BMDMs by hydrogel-treated CD4⁺ T cells.

n Representative ALP staining of PDLSCs (top) and TRAP staining of BMDMs (bottom) following co-culture with CD4⁺ T cells from indicated groups. Scale bar, 100 μm.

o Schematic mechanism illustrating that the AVD@pCuMOF/BSA-AP hydrogel restores bone homeostasis by correcting CD4⁺ T cell dysfunction and directly modulating bone cell activity.

Experimental groups were defined as follows: I, Ctrl; II, HG + LPS; III, AVD; IV, AP hydrogel; V, pCuMOF/BSA-AP hydrogel; VI, AVD@pCuMOF/BSA-AP hydrogel. Data are presented as mean ± standard deviation. n = 3 independent biological replicates per group. Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparison test. *P < 0.05 was considered statistically significant. Schematics were created using BioRender.com.

Simultaneously, we evaluated the ability of AVD@pCuMOF/BSA-AP hydrogel to arrest pathological bone resorption using RANKL-stimulated BMDMs under diabetic conditions (HG + LPS) [47]. BMDMs exhibited significantly enhanced osteoclastogenesis, characterized by the formation of numerous TRAP-positive multinucleated osteoclasts and distinct F-actin rings, indicative of active bone resorption (Fig. 6h–j). Notably, treatment with the AVD@pCuMOF/BSA-AP hydrogel attenuated these pathological osteoclastogenic changes, dramatically reducing osteoclast formation compared to control groups. This morphological improvement correlated with downregulation of key osteolytic markers, including cathepsin K (CTSK) and matrix metalloproteinase 9 (MMP9), at both mRNA and protein levels (Fig. 6k and l; Fig. S30). Together, these findings demonstrate that AVD@pCuMOF/BSA-AP hydrogel effectively counters DPD-associated bone pathology by simultaneously supporting osteogenic differentiation capacity and suppressing pathological osteoclast activation.

Finally, to determine whether AVD@pCuMOF/BSA-AP-mediated CD4⁺ T cell modulation translates into functional bone benefits, we employed a transwell co-culture system (Fig. 6m). CD4⁺ T cells conditioned in DPD medium markedly suppressed PDLSC osteogenesis (reduced ALP activity) and promoted BMDM osteoclastogenesis (increased TRAP⁺ cells). Conversely, co-culture with AVD@pCuMOF/BSA-AP hydrogel-treated CD4⁺ T cells significantly enhanced osteogenic potential and inhibited osteoclast formation, surpassing the effects of AVD alone (Fig. 6n; Fig. S31). Collectively, these results indicate that the AVD@pCuMOF/BSA-AP hydrogel exerts a comprehensive therapeutic effect through direct regulation of bone cells and modulation of the CD4⁺ T cell associated osteoimmune microenvironment, thereby offering a holistic strategy for DPD regeneration (Fig. 6o).

2.6. Therapeutic efficacy of AVD@pCuMOF/BSA-AP hydrogel in a type 2 diabetic periodontitis rat model

The in vivo therapeutic effect of AVD@pCuMOF/BSA-AP hydrogel was assessed in a rat model of DPD induced by a high-fat diet, streptozotocin, and ligature placement. Following model validation, DPD rats were randomized into 5 groups (n = 6 rats in each group), and received local periodontal injections of PBS, AP, AVD@AP, pCuMOF/BSA-AP, or AVD@pCuMOF/BSA-AP hydrogel once a week for two weeks (Fig. 7a). Age-matched healthy rats served as normal controls. Micro-CT analysis at four weeks post-treatment revealed that while PBS-treated DPD rats suffered severe alveolar bone loss, the AVD@pCuMOF/BSA-AP hydrogel partially preserved alveolar bone architecture (Fig. 7b). Quantitative analysis indicated a 41.7% reduction in cementoenamel junction to alveolar bone crest (CEJ-ABC) distance and significant improvements in bone mineral density (BMD, +27.4%) and bone volume fraction (BV/TV, +32.1%) compared to the PBS group (P < 0.001; Fig. 7d). Notably, this combination therapy demonstrated superior efficacy of AVD@pCuMOF/BSA-AP hydrogel compared to AVD monotherapy (P < 0.05), validating the synergy of multi-component design. Histological analysis (H&E staining) corroborated these findings, showing that the hydrogel maintained physiological periodontal organization and trabecular integrity, contrasting sharply with the extensive tissue collapse observed in PBS-treated controls (Fig. 7c). Consistently, Masson staining further demonstrated enhanced collagen deposition and improved periodontal matrix organization in the AVD@pCuMOF/BSA-AP group compared with the PBS group (Fig. S32).

Fig. 7.

AVD@pCuMOF/BSA-AP hydrogel promoting regeneration of alveolar bone in DPD rats.

a Timeline schematic of DPD model establishment, hydrogel treatment, and week 4 endpoint analysis for bone remodeling evaluation.

b Representative 3D micro-CT reconstructions (top) and corresponding buccal-palatal cross-sections (bottom) of the maxillary alveolar bone from different treatment groups. Scale bar, 2 mm.

c Representative H&E staining of periodontal tissues. Black lines indicate CEJ-ABC distance; red lines indicate alveolar bone crest. Scale bar, 500 μm 

d Quantitative micro-CT analysis of CEJ–ABC distance, bone mineral density (BMD), and bone volume fraction (BV/TV).

e Representative immunohistochemistry staining for osteogenic markers ALP and OPN, and immunofluorescence images of RUNX2 (green). Nuclei were stained with DAPI (blue). Red arrows indicate ALP-positive or OPN-positive cells. Scale bar, 50 μm or 100 μm.

f Quantification of ALP and OPN positive areas and RUNX2 mean fluorescence intensity.

g Representative TRAP staining images (top) and immunohistochemistry images of CTSK (bottom) in the periodontal tissues. Red arrows indicate TRAP- positive osteoclasts or CTSK-positive cells. Scale bar, 50 μm 

h Quantification of osteoclast activity by osteoclast surface per bone surface (Oc.S/BS) and osteoclast number per bone perimeter (Oc.N/B.Pm), and CTSK⁺ cell counts per field.

Experimental groups were defined as follows: I, Normal; II, PBS; III, AP hydrogel; IV, AVD@AP hydrogel; V, pCuMOF/BSA-AP hydrogel; VI, AVD@pCuMOF/BSA-AP hydrogel. Data are presented as mean ± standard deviation. n = 6 rats per group. Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparison test. *P < 0.05 was considered statistically significant. Schematics were created using BioRender.com.

To assess bone remodeling status following AVD@pCuMOF/BSA-AP treatment, key markers of bone metabolism were analyzed. Immunohistochemical staining revealed significantly enhanced expression of osteogenic markers (ALP, OPN, and COL1) in the periodontal ligament and alveolar bone of AVD@pCuMOF/BSA-AP hydrogel-treated rats compared to the PBS group (Fig. 7e and f; Fig. S33). Concurrent immunofluorescence detection of RUNX2 demonstrated markedly upregulated expression following AVD@pCuMOF/BSA-AP treatment (Fig. 7e and f). We also assessed anti-resorptive effects via TRAP staining and CTSK immunohistochemistry (Fig. 7g and h). The PBS group exhibited extensive osteoclast activity, evidenced by abundant TRAP-positive cells and CTSK-positive cells along alveolar bone surfaces. Strikingly, the AVD@pCuMOF/BSA-AP treatment nearly abolished the presence of osteoclasts, effectively reversing the osteolytic phenotype. Collectively, the AVD@pCuMOF/BSA-AP hydrogel promotes alveolar bone regeneration by simultaneously boosting osteogenesis and arresting pathological resorption.

2.7. Restoration of immune and redox homeostasis in vivo

The pathogenesis of DPD involves a self-perpetuating cycle of oxidative stress and adaptive immune dysregulation [16,48,49]. To delineate the mechanistic basis underlying the therapeutic effects of AVD@pCuMOF/BSA-AP hydrogel, we evaluated its impact on redox homeostasis and immune regulation at week 2 after treatment (Fig. 8a). Biochemical analysis demonstrated severe oxidative damage in DPD gingival tissues, characterized by elevated H₂O₂ (4.1-fold increase, P < 0.001) and lipid peroxidation (MDA, 2.8-fold, P < 0.001) alongside impaired SOD2 activity (69.1% reduction, P < 0.001; Fig. 8b). In contrast, the AVD@pCuMOF/BSA-AP treatment improved redox balance, reducing H₂O₂ and MDA levels by 72.9% and 50.2%, respectively (P < 0.001 vs. PBS group), while normalizing SOD2 activity to 94.6% of healthy levels. This redox restoration was further confirmed by DHE staining and the upregulated expression of antioxidant enzymes (SOD2, CAT) in situ (Fig. 8c and d; Fig. S34). Furthermore, neutrophil-specific Ly6G staining confirmed that the heightened respiratory burst activity in DPD tissues was markedly suppressed by the AVD@pCuMOF/BSA-AP hydrogel (P < 0.001; Fig. S35). We further explored in vivo anti-inflammatory activity by assessing inflammatory markers [33]. TNF-α, IL-1β, and IL-6 levels were significantly reduced following treatment with AP, AVD@AP, pCuMOF/BSA-AP, and AVD@pCuMOF/BSA-AP compared to the PBS group (Fig. 8e and f; Fig. S36). Particularly, the AVD@pCuMOF/BSA-AP group exhibited significantly lower cytokine levels than the AVD@AP group, highlighting the limitations of AVD monotherapy in DPD management.

Fig. 8.

AVD@pCuMOF/BSA-AP hydrogel restoring immune and redox homeostasis in vivo.

a Schematic illustration of the experimental design and analytical workflow for biochemical and immunological assessments at week 2 after treatment.

b Biochemical quantification of oxidative stress markers (H₂O₂, MDA) and antioxidant enzyme activity (SOD) in periodontal tissues. MDA, malondialdehyde.

c DHE (red) staining and immunofluorescence staining for SOD2 (green) in gingival tissues.

d Quantification of MFI for DHE and SOD2. Scale bar, 50 μm.

e Representative immunofluorescence images of TNF-α (green) and immunohistochemical staining for IL-1β in periodontal tissues. Red arrows indicate IL-1β-positive regions. Scale bar, 100 μm or 50 μm.

f Quantification of TNF-α fluorescence intensity and IL-1β expression levels.

g Flow cytometric analysis of gingival CD4⁺ T cell subsets, including Th17 (IL-17A⁺), Treg (FOXP3⁺), Th1 (IFN-γ⁺), Th2 (IL-4⁺), and exhausted T cells (PD-1⁺, CTLA4⁺).

h Quantification of the frequencies of the indicated CD4⁺ T cell subsets.

i Representative immunofluorescence images showing colocalization of CD4 (red) with FOXP3, IL-17A, or PD-1 (green) in periodontal tissues. Scale bar, 40 μm.

Experimental groups were defined as follows: I, Normal; II, PBS; III, AP hydrogel; IV, AVD@AP hydrogel; V, pCuMOF/BSA-AP hydrogel; VI, AVD@pCuMOF/BSA-AP hydrogel. Data are presented as mean ± standard deviation. n = 6 rats per group. Statistical significance was determined using one-way ANOVA followed by Tukey's multiple comparison test. *P < 0.05 was considered statistically significant. Schematics were created using BioRender.com.

Finally, we characterized CD4⁺ T cell subsets to elucidate the immunomodulatory mechanism of AVD@pCuMOF/BSA-AP hydrogel. Flow cytometry revealed that DPD induced significant disruption of T cell homeostasis, with increased Th17 (IL-17A⁺) and Th1 (IFN-γ⁺) populations, decreased Treg (FOXP3⁺) and Th2 (IL-4⁺) populations (Fig. 8g and h; Fig. S37). Remarkably, AVD@pCuMOF/BSA-AP treatment partially restored immunological balance, normalizing Th17/Treg and Th1/Th2 ratios and alleviating CD4⁺ T cell exhaustion. Immunofluorescence staining validated the robust infiltration of FOXP3⁺ Tregs and the exclusion of IL-17A⁺ Th17 cells in AVD@pCuMOF/BSA-AP hydrogel-treated tissues (Fig. 8i).

Collectively, these results demonstrate that hyperglycemia-induced redox imbalance is associated with oxidative damage in DPD [50], while the AVD@pCuMOF/BSA-AP hydrogel attenuates key pathological features of DPD, including oxidative stress and immune dysregulation. It concurrently neutralizes the inflammatory microenvironment and modulates adaptive immunity, thereby creating a permissive microenvironment for periodontal tissue repair. Comprehensive safety evaluations confirmed the excellent biocompatibility of the system (Fig. S38), supporting its translational potential. Future studies incorporating multiple time points will further clarify the dynamic relationship between early immune regulation and subsequent bone remodeling.

3. Conclusion

In summary, we have engineered a microenvironment-responsive hydrogel-nanoparticle hybrid system designed that spatiotemporally reprograms the osteoimmune environment for DPD treatment. By incorporating pathological cue-responsive properties into the hydrogel architecture, we established a hierarchical therapeutic sequence driven by ROS-logic actuation. In the early phase, ROS-triggered matrix disassembly enables simultaneous oxidative stress scavenging and targeted release of antibody-functionalized BSA nanoparticles, facilitating intracellular immunomodulator delivery. This intervention restores CD4⁺ T-cell immune homeostasis through mitochondrial quality control and metabolic reprogramming, thereby attenuating exhaustion-associated dysfunction and interrupting redox-driven immune imbalance. As inflammation resolves, the platform transitions into a regenerative phase in which polyphenol-engineered MOF depots provide sustained release of AVD and osteogenic Cu²⁺, coupling long-term immunomodulation with coordinated bone remodeling.

Collectively, this logic-gated, hierarchical delivery strategy effectively improves adaptive immune balance and re-establishes bone metabolic balance, leading to effective alveolar bone regeneration in diabetic models. By integrating redox logic actuation, targeted immunometabolic regulation, and sustained osteomodulation within a single platform, this work provides a potential therapeutic strategy for inflammatory bone repair. Beyond diabetic periodontitis, such spatiotemporally programmed metabolic intervention strategies may offer broad translational potential for chronic inflammatory osteopathies.

CRediT authorship contribution statement

Yujun Jiang: Writing – original draft, Visualization, Methodology, Investigation, Data curation, Conceptualization. Shuyun Liu: Writing – original draft, Visualization, Methodology, Investigation, Conceptualization. Xiaolin Li: Methodology, Investigation. Lu Han: Writing – review & editing, Supervision, Resources, Funding acquisition, Conceptualization. Hongrui Liu: Writing – review & editing, Supervision, Funding acquisition. Minqi Li: Supervision, Resources, Funding acquisition.

Data availability

The mouse gingival scRNA-seq datasets analyzed in this study were retrieved from the NCBI Gene Expression Omnibus (GEO) under accession numbers GSE188217. The mouse spleen RNA-seq datasets analyzed in this study were retrieved from the GEO database under accession numbers GSE267853.

All data supporting the conclusions in the paper are present in the paper and/or the Supplementary Information. All data underlying this study are available from the corresponding author upon request.

Ethics approval and consent to participate

All animal experiments were approved by the Ethics Committee of Shandong University School of Stomatology (No. 20240103), and followed the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines for preclinical animal studies.

Human gingival samples collection and cell isolation were approved by the Ethics Committee of Shandong University School of Stomatology (No. 20240331). The study design and conduct complied with the Department of Health and Human Services, the Declaration of Helsinki, and all relevant ethical regulations. Written, informed consent was provided by all patients before enrollment.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Appendix A. Supplementary data

The following is the Supplementary data to this article: