Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo therapeutic effect of Mn-NC composite hydrogels on TMJ-OA after 2 weeks. (a) Schematic diagram of in vivo experiments of hydrogel treatment of TMJ-OA. HE staining (b), SO/FG staining (c), immunohistochemical staining of COLII (d) and MMP13 (e), immunofluorescence staining of CD86 (f) and CD206 (g) at the synovial, and immunofluorescence staining of IL-1β (h) in the TMJ of rats after 2 weeks of treatment, scale bar = 100 μm. (i) Histological OARSI score of articular cartilage after 2 weeks of treatment and quantitative data of immunohistochemical positive area and proportion of immunofluorescence positive cells in (c–h). Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo therapeutic effect of Mn-NC composite hydrogels on TMJ-OA after 8 weeks. HE staining (a), SO/FG staining (b), immunohistochemical staining of COLII (c) and MMP13 (d), immunofluorescence staining of CD86 (e) and CD206 (f) at the synovial, and immunofluorescence staining of IL-1β (g) in the TMJ of rats after 8 weeks of treatment, scale bar = 100 μm. (h) Histological OARSI score of articular cartilage after 8 weeks of treatment and quantitative data of immunohistochemical positive area and proportion of immunofluorescence positive cells in (b–g). Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunofluorescence, Comparison

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: (A) Galectin-family protein groups, including an LGALS13-annotated / CLC-Galectin-10-like porcine orthologous protein group. (B) Immune lineage-associated proteins (T-cell, B-cell, myeloid, cytotoxic, and pan-leukocyte markers). (C) Selected signaling protein groups (NF-κB and JAK/STAT axes). DIA proteomics was performed on effluent cell pellets from one representative biological perfusion in technical triplicate and should be interpreted as exploratory cell-associated protein abundance. Values are log2 fold-changes relative to the Pre-1 baseline; heatmap scale capped at ±3 for display. UD = undetected.

Article Snippet: An acellular intact-spleen perfusion platform enables serial cellular, transcriptomic, proteomic, and functional pharmacodynamic sampling that identifies steroid-responsive transcriptional programs, validates SOCS3 protein induction, and nominates a CLC/Galectin-10-like non-transcriptional immunosuppressive axis for translation to discarded human donor spleens.

Techniques: Quantitative Proteomics

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: An acellular intact-spleen perfusion platform enables serial cellular, transcriptomic, proteomic, and functional pharmacodynamic sampling that identifies steroid-responsive transcriptional programs, validates SOCS3 protein induction, and nominates a CLC/Galectin-10-like non-transcriptional immunosuppressive axis for translation to discarded human donor spleens.

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: (A) Galectin-family protein groups, including an LGALS13-annotated / CLC-Galectin-10-like porcine orthologous protein group. (B) Immune lineage-associated proteins (T-cell, B-cell, myeloid, cytotoxic, and pan-leukocyte markers). (C) Selected signaling protein groups (NF-κB and JAK/STAT axes). DIA proteomics was performed on effluent cell pellets from one representative biological perfusion in technical triplicate and should be interpreted as exploratory cell-associated protein abundance. Values are log2 fold-changes relative to the Pre-1 baseline; heatmap scale capped at ±3 for display. UD = undetected.

Article Snippet: The assay was designed as an orthogonal functional test of the human immune counterpart of the porcine LGALS13-annotated / CLC-Galectin-10-like proteomic signal.

Techniques: Quantitative Proteomics

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: The assay was designed as an orthogonal functional test of the human immune counterpart of the porcine LGALS13-annotated / CLC-Galectin-10-like proteomic signal.

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: (A) Galectin-family protein groups, including an LGALS13-annotated / CLC-Galectin-10-like porcine orthologous protein group. (B) Immune lineage-associated proteins (T-cell, B-cell, myeloid, cytotoxic, and pan-leukocyte markers). (C) Selected signaling protein groups (NF-κB and JAK/STAT axes). DIA proteomics was performed on effluent cell pellets from one representative biological perfusion in technical triplicate and should be interpreted as exploratory cell-associated protein abundance. Values are log2 fold-changes relative to the Pre-1 baseline; heatmap scale capped at ±3 for display. UD = undetected.

Article Snippet: In parallel, data independent acquisition (DIA) proteomics of effluent cell pellets nominated a non-transcriptional protein-level response: a Sus scrofa LGALS13-annotated, CLC/Galectin-10-like galectin detected despite absence of the corresponding effluent-cell transcript.

Techniques: Quantitative Proteomics

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: In parallel, data independent acquisition (DIA) proteomics of effluent cell pellets nominated a non-transcriptional protein-level response: a Sus scrofa LGALS13-annotated, CLC/Galectin-10-like galectin detected despite absence of the corresponding effluent-cell transcript.

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced