Journal: Bioactive Materials

Article Title: Tailoring nanotopography and antibacterial properties of calcium phosphate bone grafts via fluoride incorporation

doi: 10.1016/j.bioactmat.2025.12.026

Figure Lengend Snippet: Bacterial viability of P. aeruginosa (a) and S. aureus (b). i) Quantitative analysis of the % of dead/total bacteria at 6 h incubation on the different CaP discs obtained by hydrolysis of α-TCP in sodium fluoride solutions (0 mM [0F], 0.5 mM [05F], and 10 mM [10F]) either in biomimetic (BM) or hydrothermal (AC) conditions and merged CLSM images of live/dead staining for representative samples. As controls, Flat and re-pressed fluoride-doped CaP nanostructures (CXF_AC or CXF_BM) were used. Red bacteria correspond to bacteria with damaged membrane, green bacteria correspond to viable bacteria. ii) BacTiterGlo measurements normalized with the area of bacteria adhered at 6 h of incubation on the CaP substrates. The red dotted line indicates the normalized metabolic activity of a Ti control. iii) SEM images of P. aeruginosa and S. aureus cultured onto different CaP discs. n = 3; ns, ∗, ∗∗, ∗∗∗ and ∗∗∗∗ indicate significance at p > 0.05, p ≤ 0.05, p ≤ 0.01, p ≤ 0.001 and p ≤ 0.0001, respectively.

Article Snippet: Bacterial assays were conducted using two strains: Gram-negative P. aeruginosa (ATCC 27853) and Gram-positive S. aureus (ATCC 25923).

Techniques: Bacteria, Incubation, Staining, Membrane, Activity Assay, Control, Cell Culture

Journal: Bioactive Materials

Article Title: Tailoring nanotopography and antibacterial properties of calcium phosphate bone grafts via fluoride incorporation

doi: 10.1016/j.bioactmat.2025.12.026

Figure Lengend Snippet: Co-culture of P. aeruginosa and SaOS-2 cells on the nanostructured CaP discs, as well as Flat and Ti controls. a) Merged CLSM images of: i) a pre-implantation infection model, where the samples were first incubated for 6 h with P. aeruginosa and subsequently SaOS-2 cells were cultured for 24 h; or ii) post-implantation infection model, where SaOS-2 cells were first cultured for 24 h on the substrates, which were subsequently incubated for 6 h with P. aeruginosa . Actin filaments were stained with Alexa Fluor™ 546 phalloidin (orange fluorescence signal) and the nuclei with DAPI (blue fluorescence signal). (b) Orthogonal CLSM images showing simultaneous co-visualization of cells and bacteria stained with Alexa Fluor™ 546 phalloidin (orange fluorescence signal), the nuclei with DAPI (blue fluorescence signal) and SYTO-9 (green fluorescence signal). (c) Dead percentage of P. aeruginosa onto the different CaP substrates and the controls, for both the pre-implantation infection i) and the post-implantation ii) infection models. n = 3; ns and ∗ indicate significance at p > 0.05 and p ≤ 0.05, respectively.

Article Snippet: Bacterial assays were conducted using two strains: Gram-negative P. aeruginosa (ATCC 27853) and Gram-positive S. aureus (ATCC 25923).

Techniques: Co-Culture Assay, Infection, Incubation, Cell Culture, Staining, Fluorescence, Bacteria

Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can activate the PI3K/AKT pathway by inhibiting the expression of PTEN in glioma cells.Western blotting was used to detect the expression of PTEN in U87 and U251 cell lines (A); the CCK-8 assay was used to measure the viability of U87 and U251 glioma cells treated with different concentrations of LY294002 and 740 Y-P (B); Western blotting was also used to detect the expression of PI3K, p-AKT/AKT in U87 and U251 cell lines (C).*P < 0.05 vs si-NC，@P < 0.05 vs si-NC+Agonist，#P < 0.05 vs si-TNKS1，&P < 0.05 vs OE-TNKS1.

Article Snippet: The cells were then cultured in the incubator for another 12 h. Subsequently, the cells in each well were randomly divided into groups: the si-NC+agonist group, the si-TNKS1 +agonist group, and the OE-TNKS1 +inhibitor group were treated with PI3K agonist 740 Y-P (HY-P0175, MCE, USA) and PI3K inhibitor LY294002 (HY-10108, MCE, USA) and continued to be cultured for 24 h. The TNKS1-siRNA empty vector group (si-NC), the TNKS1-siRNA group (si-TNKS1), and the TNKS1 overexpression group (OE-TNKS1) were treated with an equivalent amount of diluent and continued to be cultured for 24 h.

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can regulate the proliferation and glycolysis of glioma cells by mediating the PTEN-PI3K/AKT pathway.Cell viability of various groups in U87 and U251 cell lines (A); detection of glucose (GLU) and lactate (LA) levels in the culture supernatant of various groups in U87 and U251 cell lines using biochemical assay kits (B); qPCR was used to detect the mRNA levels of GLUT1 and HK2 in various groups of U87 and U251 cell lines (C). Statistical significance *p < 0.05, **p < 0.01, ***p < 0.001, **** p < 0.0001,"ns" P ≥ 0.05.

Article Snippet: The cells were then cultured in the incubator for another 12 h. Subsequently, the cells in each well were randomly divided into groups: the si-NC+agonist group, the si-TNKS1 +agonist group, and the OE-TNKS1 +inhibitor group were treated with PI3K agonist 740 Y-P (HY-P0175, MCE, USA) and PI3K inhibitor LY294002 (HY-10108, MCE, USA) and continued to be cultured for 24 h. The TNKS1-siRNA empty vector group (si-NC), the TNKS1-siRNA group (si-TNKS1), and the TNKS1 overexpression group (OE-TNKS1) were treated with an equivalent amount of diluent and continued to be cultured for 24 h.

Techniques:

Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can regulate the PTEN-PI3K/AKT pathway in glioma xenografts.Western blotting was used to detect the protein expression levels of PTEN, PI3K, p-AKT, and AKT in cells from each group.* P < 0.05 vs TNKS1-siRNA NC, @ P < 0.05 vs TNKS1-siRNA，# P < 0.05 vs Inhibitor.

Article Snippet: The cells were then cultured in the incubator for another 12 h. Subsequently, the cells in each well were randomly divided into groups: the si-NC+agonist group, the si-TNKS1 +agonist group, and the OE-TNKS1 +inhibitor group were treated with PI3K agonist 740 Y-P (HY-P0175, MCE, USA) and PI3K inhibitor LY294002 (HY-10108, MCE, USA) and continued to be cultured for 24 h. The TNKS1-siRNA empty vector group (si-NC), the TNKS1-siRNA group (si-TNKS1), and the TNKS1 overexpression group (OE-TNKS1) were treated with an equivalent amount of diluent and continued to be cultured for 24 h.

Techniques: Western Blot, Expressing

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs restores Hippo signaling balance by inhibiting LATS1 and YAP phosphorylation. (A) Volcano plot of RNA-sequencing analysis for H 2 O 2 -stimulated mouse NPCs with or without NM-LP TK /RSV-MnCDs treatment. (B) KEGG enrichment analysis of RNA-sequencing analysis. (C) Animal grouping and corresponding treatments. (D–G) IF images and quantitative analyses of p-LATS1 and p-YAP in NP tissues from various groups (n = 5). Scale bars, 100 μm. (H) In vitro experiments grouping and corresponding treatments. (I–L) IF images and quantitative analyses of p-LATS1 and p-YAP in NPCs (n = 3). Scale bars, 20 μm. (M–N) IF images and quantitative analyses of total YAP in NPCs (n = 3). Scale bars, 20 μm. (O–Q) Western blot results and corresponding quantification analyses for YAP in nucleus and p-YAP in cytoplasm (n = 3). (R–S) Docking verification of RSV and human and mouse LATS1. Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Phospho-proteomics, RNA Sequencing, In Vitro, Western Blot

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs suppressed oxidative stress and aberrant neurovascularization in a Hippo pathway suppression–dependent manner. (A) Representative IF images and 3D surface plot analyses via ImageJ for p-LATS1 in NPCs treated with H 2 O 2 , MCB, TRULI, NM-LP TK /RSV-MnCDs, or their combinations (n = 3). Scale bar, 20 μm. (B) Grouping illustration of the treatment strategy. (C) Representative IF images and 3D surface plot analyses via ImageJ for p-YAP. (D–F) Mitochondrial respiration function measured by Seahorse XF analyzer (n = 3). (G) Intracellular ROS levels assessed by fluorescent ROS probes (n = 3). Scale bar, 50 μm. (H) Representative tube formation images of MAECs co-cultured with NPCs (n = 5). Scale bar, 200 μm. (I–J) Quantitative analysis of capillary branch points and tube length (n = 5). (K–L) TUBB3-labed neurofilaments of DRG neurons and quantification analysis for length (n = 5). Scale bar, 20 μm. (M–P) Semi-quantification analysis for mRNA levels of Sema3a , Slit2 , Netrin-1 , and Vegfa (n = 3). Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Cell Culture

Journal: Bioactive Materials

Article Title: Mitochondrial-targeted injectable hydrogel for periodontitis therapy via oral immunity and flora regulation

doi: 10.1016/j.bioactmat.2025.12.002

Figure Lengend Snippet: Study on antibacterial and antibiofilm activity, and mechanism of hydrogels in vitro . (A) Representative images of the colonies of S. aureus , MRSA , E. coli and P. gingivalis on LB agar plates and (B, C) their relative bacterial viability in different groups (n = 3). (D) SEM images of MRSA and P. gingivalis . (E) Live (green)/dead (red) staining of MRSA and P. gingivalis , and (F) corresponding quantitative analysis of the live/dead ratio (n = 3). (G) The diagram of antimicrobial mechanism of melittin. (H) Crystal violet staining of the bacterial biofilms and corresponding quantitative analysis of (I, J) biofilm formation and (K, L) biofilm reduction in different groups (n = 3). (M) Membrane permeabilizing and (N) depolarization effect of hydrogel against MRSA and P. ginivalis . (O) Protein leakage in MRSA and P. ginivalis after hydrogels treatment (n = 3). (P) Metabolic activity of MRSA and P. ginivalis under different treatments (n = 3). ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: The antibacterial efficacy of hydrogels was evaluated using S. aureus (ATCC 6538), P. gingivalis (ATCC 33277), E. coli (ATCC 8739), and MRSA (ATCC 43300).

Techniques: Activity Assay, In Vitro, Staining, Membrane