Journal: Bioactive Materials

Article Title: Integrated apoptotic extracellular vesicle-recruitment peptide coating reprograms the diabetic bone microenvironment and orchestrates enhanced implant osseointegration

doi: 10.1016/j.bioactmat.2026.05.059

Figure Lengend Snippet: In vivo immunomodulatory effects of different modified surfaces in a DM model. (A) Schematic representation of the animal modeling and experimental treatment workflow. (B, C) hematoxylin and eosin staining of the peri-implant tissues in the femurs of DM rats 1 week after implantation, accompanied by quantitative analysis of the fibrous capsule thickness (scale bar = 100 μm, n = 5). (D–G) Immunofluorescence staining evaluating the polarization state of macrophages surrounding the implants (green: macrophage marker cluster of differentiation (CD) 68; red: M1 marker CD86 and M2 marker CD206; blue: nuclei), along with corresponding quantitative analysis of the fluorescence signals (scale bar = 100 μm, n = 5). (H–K) Immunohistochemical staining assessing the expression of the pro-inflammatory marker tumor necrosis factor-α and the anti-inflammatory marker interleukin-10 in the peri-implant area, with quantitative results of the positive staining areas (scale bar = 100 μm, n = 5). Data are expressed as the mean ± standard deviation, with statistical analysis performed using one-way ANOVA and Tukey's post-hoc test. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicate statistical significance.

Article Snippet: For IHC analysis, sections underwent heat-induced antigen retrieval and blocking prior to incubation with antibodies against TNF-α (GB11188, Servicebio, China), IL-10 (GB11534, Servicebio, China), and VEGF (GB15165, Servicebio, China) to identify differences in local inflammatory and angiogenic profiles.

Techniques: In Vivo, Modification, Staining, Immunofluorescence, Marker, Fluorescence, Immunohistochemical staining, Expressing, Standard Deviation

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Systemic TREM1 ablation attenuates neuroinflammation and neurological deficits after TBI. (a) Schematic timeline of the experimental design. WT and Trem1 −/− mice were subjected to CCI or sham surgery, followed by longitudinal behavioral and histological assessments. (b, c) Analysis of the neuroinflammatory profile in the perilesional cortex at 3 days post‐TBI. (b) Representative images of inflammatory cytokines in TBI lesions detected by the Proteome Profiler Array Mouse Cytokine Array Panel A in WT and Trem1 −/− mice at 3 days post‐TBI. (c) Quantification of inflammatory cytokines in TBI lesions of WT and Trem1 −/− mice at 3 days post‐TBI or sham treatment by ELISA ( n = 5 mice per group, two‐way ANOVA followed by Tukey's multiple‐comparison test). (d) Representative immunofluorescence images of macrophages (IBA1 + , green) and the reactivation marker CD86 (red) in the TBI cortex. Trem1 ablation significantly reduced reactive polarization (scale bar, 50 µm). (e) Evaluation of neuronal apoptosis. Representative co‐staining of TUNEL (green) and NeuN (red) (left) ( n = 3 mice per group, scale bar, 100 µm). Yellow arrows indicate apoptotic neurons. (f) Assessment of white matter integrity. Representative images of Myelin Basic Protein (MBP, green) and NeuN (red) (left) and quantification of MBP fluorescence intensity (right) (scale bar, 100 µm). (g) Representative immunofluorescence images of hippocampal neurons (NeuN, red) showing preserved cellular density in Trem1 −/− mice ( n = 3 mice per group; scale bar, 100 µm). (h) Brain water content analysis at 1day and 3 d post‐TBI, indicating alleviated cerebral edema in Trem1 −/− mice ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (i, j) Assessment of blood‐brain barrier (BBB) permeability. (i) Representative images of Evans Blue extravasation. (j) Quantification of dye leakage ( n = 6 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (k, l) Molecular validation of tissue preservation. (k) Representative Western blots of the neuronal marker beta3‐tubulin, synaptic marker Synaptophysin (Syn), and tight junction protein Occludin. l, Densitometric quantification normalized to GAPDH ( n = 3 independent experimental replicates; two‐way ANOVA followed by Tukey's multiple‐comparison test). (m–o) Assessment of sensorimotor function. m, Latency to fall in the accelerating Rotarod test ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (n) Performance in the fixed‐speed (30 rpm) Rotarod test ( n = 8 mice per group, one‐way ANOVA followed by Tukey's multiple‐comparison test). o, Modified Neurological Severity Score (mNSS) trajectory ( n = 8 mice per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). (p, q) Evaluation of anxiety‐like behavior and exploratory activity in the Open Field Test (OFT). (p) Representative locomotor tracking plots. (q) Quantification of the percentage of time spent in the center zone ( n = 8 per group; two‐way ANOVA followed by Tukey's multiple‐comparison test). Data are presented as mean ± SD. Each dot represents an individual mouse, except in l, where each dot represents an independent experimental replicate. Statistical tests are specified for each panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: TNFα ELISA kit , ABclonal , # RK00027.

Techniques: Analysis, Enzyme-linked Immunosorbent Assay, Comparison, Immunofluorescence, Marker, Staining, TUNEL Assay, Fluorescence, Permeability, Biomarker Discovery, Tissue, Preserving, Western Blot, Modification, Activity Assay

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Generative AI‐driven design and mechanistic validation of a TREM1‐targeting LYTAC degrader. (a) Schematic of the RFdiffusion‐guided pipeline for de novo TREM1‐binding peptide design. Peptide backbones were generated around the Ig‐like domain of TREM1 and subsequently screened by AlphaFold3, ZDOCK, HDOCK, and Rosetta‐based energetic analysis to identify high‐ranking candidates. (b) Predicted structural models of Pep15, Pep17, and Pep457 in complex with TREM1, showing the predicted binding interfaces. (c) Temperature‐related intensity change (TRIC) assays showing concentration‐dependent target engagement of Pep15, Pep17, and Pep457 with TREM1. Left, schematic illustration of the TRIC assay principle; right, dose‐response curves of the indicated peptides ( n = 3 independent experiments). (d) ELISA‐based binding analysis of Pep15, Pep17, and Pep457 to immobilized TREM1. Top, schematic of the ELISA workflow; bottom, quantitative binding curves ( n = 3 independent experiments). (e) Surface plasmon resonance (SPR) analysis of Pep457 binding to TREM1. Top, schematic of the SPR assay; bottom, representative sensorgrams obtained at the indicated Pep457 concentrations. (f) Structural model of the bifunctional TREM1‐LYTAC, in which Pep457 engages TREM1 and the mannose‐6‐phosphate (M6P) moiety recruits the cation‐independent mannose‐6‐phosphate receptor (CI‐M6PR). (g) Representative confocal images showing partial colocalization of TREM1‐LYTAC with lysosomes following cellular uptake. TREM1‐LYTAC, green; lysosomes, red; nuclei, blue. Arrows indicate representative colocalized puncta. Scale bars as indicated. (h) ELISA quantification of cellular TREM1 protein following treatment with vehicle, scrambled‐LYTAC, TREM1‐LYTAC, or TREM1‐LYTAC in the presence of bafilomycin A1 (BafA1) or MG132 ( n = 5 independent experiments). (i) Representative flow‐cytometric histograms showing cell‐surface TREM1 expression after the indicated treatments. (j) Trem1 mRNA expression measured by qPCR after the indicated treatments ( n = 3 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (k) Requirement of both functional modules for TREM1 reduction. Cellular TREM1 protein levels were quantified after treatment with Pep457 plus M6P, Pep457 alone, M6P alone, TREM1‐LYTAC in the presence of CI‐M6PR blockade, or conjugated TREM1‐LYTAC ( n = 5 independent experiments; one‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Plasma pharmacokinetic profile of free TREM1‐LYTAC after intravenous administration. Fluorescence intensity was fitted to a monoexponential decay model, yielding an apparent half‐life of 1.13 h ( n = 3 mice). (m) Proposed mechanism of TREM1‐LYTAC‐mediated degradation. TREM1‐LYTAC bridges cell‐surface TREM1 to CI‐M6PR, thereby promoting receptor internalization and lysosomal degradation. Data are mean ± SD. Each dot represents an independent experimental replicate or an individual animal. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: TNFα ELISA kit , ABclonal , # RK00027.

Techniques: Biomarker Discovery, Binding Assay, Generated, Analysis, Concentration Assay, Drug discovery, Enzyme-linked Immunosorbent Assay, SPR Assay, Confocal, Expressing, Comparison, Functional Assay, Clinical Proteomics, Fluorescence

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Engineering and validation of a BBB‐targeted, pH‐responsive nanodevice for spatiotemporal delivery of TREM1‐LYTAC. (a) Schematic illustration of APTL‐NP fabrication. TREM1‐LYTAC was conjugated to a generation‐4 PAMAM dendrimer through a terephthalaldehyde‐derived acid‐labile linker, followed by surface functionalization with Angiopep‐2 and PS‐PEG‐NHS to enable LRP1‐directed BBB targeting and improve systemic stability. (b) Representative transmission electron microscopy images of PAMAM‐based precursor particles and APTL‐NP (Scale bars, 1 µm). (c) Dynamic light scattering profiles showing the apparent hydrodynamic size distributions of G4 PAMAM and APTL‐NP under the indicated measurement conditions. (d) Time‐dependent changes in hydrodynamic diameter and polydispersity index of APTL‐NP incubated in PBS at pH 7.4 and 37°C. (e) pH‐dependent zeta‐potential profile of APTL‐NP. (f) Nanoparticle tracking analysis showing pH‐dependent particle size and concentration distributions after incubation at pH 7.5, 7.0, or 6.5. (g) pH‐responsive release of TREM1‐binding‐competent TREM1‐LYTAC from APTL‐NP. Released free TREM1‐LYTAC was separated by ultrafiltration and quantified using a TREM1‐binding ELISA with free TREM1‐LYTAC standards ( n = 3 independent experiments). (h) Representative EdU staining images of BMECs, HT22 cells, and NHAs treated with vehicle or APTL‐NP. (i) Quantification of EdU‐positive cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (j) Representative flow‐cytometric plots for apoptosis analysis in BMECs, HT22 cells, and NHAs after treatment with vehicle or APTL‐NP. (k) Quantification of apoptotic cells in the indicated cell types( n = 6 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (l) Cell viability after 24 h or 48 h exposure to the indicated concentrations of APTL‐NP ( n = 3 independent experiments, two‐way ANOVA followed by Tukey's multiple‐comparison test). (m) Plasma pharmacokinetic profile of APTL‐NP after intravenous administration. The fluorescence signal was fitted to a monoexponential decay model, yielding an apparent half‐life of 8.38 h ( n = 3 mice). (n) Representative photographs and quantification of hemolysis induced by the indicated concentrations of APTL‐NP ( n = 3 independent experiments, one‐way ANOVA followed by Tukey's multiple‐comparison test). (o, p) Plasma C3a and C5a concentrations at the indicated time points after administration of vehicle or APTL‐NP ( n = 6 mice per group, Two‐way ANOVA followed by Tukey's multiple‐comparison test). (q) Representative H&E‐stained sections of the spleen, heart, lung, liver, and kidney from vehicle‐ and APTL‐NP‐treated mice (Scale bar, 100 µm). Data are presented as mean ± SD where applicable. For in vitro assays, n denotes independent experimental replicates; for in vivo analyses, n denotes individual mice. Statistical tests are specified for each quantitative panel above. Exact p values are provided in the Source Data. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: TNFα ELISA kit , ABclonal , # RK00027.

Techniques: Biomarker Discovery, Derivative Assay, Stability, Transmission Assay, Electron Microscopy, Dynamic Light Scattering Assay, Incubation, Zeta Potential Analyzer, Analysis, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Staining, Comparison, Clinical Proteomics, Fluorescence, In Vitro, In Vivo

Journal: Advanced Science

Article Title: AI‐Designed TREM1‐Targeted LYTAC Nanoparticles Reprogram the Neuroimmune Microenvironment in Traumatic Brain Injury

doi: 10.1002/advs.77972

Figure Lengend Snippet: Key resources table.

Article Snippet: TNFα ELISA kit , ABclonal , # RK00027.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software

Journal: Journal of Cellular and Molecular Medicine

Article Title: Downregulating CHI3L2 via the STAT3 Pathway: The Mechanism of Calcitriol in Suppressing Psoriatic Inflammation and Keratinocyte Hyperproliferation

doi: 10.1111/jcmm.71367

Figure Lengend Snippet: Topical calcitriol application ameliorates IMQ‐induced psoriasis in mice. (A) Schematic illustration of the unilateral ear model experimental design. (B) Serum calcium levels. (C) Body weight changes. (D) Clinical severity scores (erythema, scaling and thickness) of the right ear, assessed on indicated days using a 0–4 scale. Cumulative scores are presented as mean ± SD. (E) Representative photographs of the right ear at endpoint. (F) Haematoxylin and eosin (H&E) staining of right ear sections. Scale bar = 100 μm. (G) Relative mRNA expression levels of IL‐1α, IL‐6, IL‐17A and IL‐23A . Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: To mimic psoriatic keratinocyte dysfunction, HaCaT cells were incubated with 10 ng/mL M5 (IL‐17A, IL‐22, IL‐1α, oncostatin M and TNF‐α) (ABclonal Technology Co. Ltd., Wuhan, China) for 24 h as reported previously [ , , , ].

Techniques: Staining, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Downregulating CHI3L2 via the STAT3 Pathway: The Mechanism of Calcitriol in Suppressing Psoriatic Inflammation and Keratinocyte Hyperproliferation

doi: 10.1111/jcmm.71367

Figure Lengend Snippet: Topical calcitriol treatment ameliorates distant psoriatic skin lesions in the bilateral ear model. (A) Schematic illustration of the bilateral ear model experimental design. (B) Serum calcium levels. (C) Body weight changes. (D) Clinical severity scores (erythema, scaling and thickness) of the right ear, assessed on indicated days using a 0–4 scale. Cumulative scores are presented as mean ± SD. (E) Representative photographs of the left ear at endpoint. (F) Haematoxylin and eosin (H&E) staining of left ear sections. Scale bar = 100 μm. (G) Relative mRNA expression levels of IL‐1α, IL‐6, IL‐17A and IL‐23A . Data are presented as mean ± SD. ** p < 0.01, **** p < 0.0001.

Article Snippet: To mimic psoriatic keratinocyte dysfunction, HaCaT cells were incubated with 10 ng/mL M5 (IL‐17A, IL‐22, IL‐1α, oncostatin M and TNF‐α) (ABclonal Technology Co. Ltd., Wuhan, China) for 24 h as reported previously [ , , , ].

Techniques: Staining, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Downregulating CHI3L2 via the STAT3 Pathway: The Mechanism of Calcitriol in Suppressing Psoriatic Inflammation and Keratinocyte Hyperproliferation

doi: 10.1111/jcmm.71367

Figure Lengend Snippet: Calcitriol ameliorates M5‐induced abnormal proliferation, migration and inflammatory responses in HaCaT cells. (A) Protein expression levels of KRT1 and KRT6 were determined by Western blot analysis, with β‐Actin serving as the internal loading control. (B) Cell viability was assessed in different treatment groups. (C) Colony formation assay was performed to evaluate the proliferative capacity of cells in each group. (D) Representative images of wound healing assay at 0 and 24 h post‐treatment. Scale bar = 400 μm. (E) Relative mRNA expression levels of IL‐1α, IL‐6, IL‐17A and IL‐23A . Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: To mimic psoriatic keratinocyte dysfunction, HaCaT cells were incubated with 10 ng/mL M5 (IL‐17A, IL‐22, IL‐1α, oncostatin M and TNF‐α) (ABclonal Technology Co. Ltd., Wuhan, China) for 24 h as reported previously [ , , , ].

Techniques: Migration, Expressing, Western Blot, Control, Colony Assay, Wound Healing Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Downregulating CHI3L2 via the STAT3 Pathway: The Mechanism of Calcitriol in Suppressing Psoriatic Inflammation and Keratinocyte Hyperproliferation

doi: 10.1111/jcmm.71367

Figure Lengend Snippet: Calcitriol ameliorates M5‐induced psoriasis‐like phenotypes through CHI3L2 downregulation. (A) The mRNA expression of RPL36A, PPBP, UBE2F, MGC32805, IL4I1, LINC00519, CHI3L2, ABHD14A and PCDHGB7 , validated by RT‐qPCR assay. (B) Efficacy of shRNA‐mediated CHI3L2 knockdown confirmed by RT‐qPCR. (C) Protein expression levels of KRT1 and KRT6 were determined by Western blot analysis, with β‐Actin serving as the internal loading control. (D) Cell viability was assessed in different treatment groups. (E) Representative images of wound healing assay at 0 and 24 h post‐treatment. Scale bar = 400 μm. (F) Relative mRNA expression levels of IL‐1α, IL‐6, IL‐17A and IL‐23A . Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: To mimic psoriatic keratinocyte dysfunction, HaCaT cells were incubated with 10 ng/mL M5 (IL‐17A, IL‐22, IL‐1α, oncostatin M and TNF‐α) (ABclonal Technology Co. Ltd., Wuhan, China) for 24 h as reported previously [ , , , ].

Techniques: Expressing, Quantitative RT-PCR, shRNA, Knockdown, Western Blot, Control, Wound Healing Assay

Journal: Molecules

Article Title: Carboxyl Amidation of Chlorogenic Acid Improves Anti-Inflammatory Activity and Biosafety via Potent AKR1B1 Inhibition in LPS-Induced Macrophages

doi: 10.3390/molecules31183247

Figure Lengend Snippet: Quantitative detection of LPS-induced inflammatory mediators after treatment with CGAA, CGA, CGL and positive control epalrestat in RAW264.7 macrophages. ( a , b ) Determination of NO production: ( a ) Full concentration–response profiles at 10–100 μM. ( b ) Single-dose comparison at 100 μM. ( c , d ) Quantification of IL-6 secretion: ( c ) Full concentration–response profiles at 10–100 μM. ( d ) Single-dose comparison at 100 μM. ( e , f ) Quantification of TNF-α secretion: ( e ) Full concentration–response profiles at 10–100 μM. ( f ) Single-dose comparison at 100 μM. Cells were pretreated with the indicated compounds (10–100 μM) for 2 h, followed by LPS (1 μg/mL) stimulation for 24 h. NO levels were determined via the Griess assay, while IL-6 and TNF-α concentrations were quantified by ELISA. All data are expressed as mean ± SD ( n = 3). #### p < 0.0001 vs. the control group; **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 vs. the LPS group.

Article Snippet: The levels of TNF-α and IL-6 in culture supernatants were quantified using commercial ELISA kits (Mouse TNF-alpha Fast Pro ELISA Kit, Cat. No. RK05228; Mouse IL-6 Fast Pro ELISA Kit, Cat. No. RK05213; ABclonal, Wuhan, China).

Techniques: Positive Control, Concentration Assay, Comparison, Griess Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecules

Article Title: Carboxyl Amidation of Chlorogenic Acid Improves Anti-Inflammatory Activity and Biosafety via Potent AKR1B1 Inhibition in LPS-Induced Macrophages

doi: 10.3390/molecules31183247

Figure Lengend Snippet: Temperature-gradient CETSA analysis of AKR1B1. Cell lysates pretreated with CGA, CGL, or CGAA were heated at 4, 45, 60, 75, or 90 °C. Residual soluble AKR1B1 protein was quantified by ELISA and normalized to the corresponding 4 °C sample. Data are presented as mean ± SD ( n = 3). **** p < 0.0001 vs. the CGA group.

Article Snippet: The levels of TNF-α and IL-6 in culture supernatants were quantified using commercial ELISA kits (Mouse TNF-alpha Fast Pro ELISA Kit, Cat. No. RK05228; Mouse IL-6 Fast Pro ELISA Kit, Cat. No. RK05213; ABclonal, Wuhan, China).

Techniques: Analysis, Enzyme-linked Immunosorbent Assay