Journal: Materials Today Bio

Article Title: Self-powered intracellular nanogenerator attenuates inflammatory osteolysis through mitochondrial MRS2/Mg 2+ -mediated macrophage repolarization and osteoclastogenesis inhibition

doi: 10.1016/j.mtbio.2025.102050

Figure Lengend Snippet: pBTO inhibited osteoclastogenesis in vitro. (A) TRAP staining in RANKL-treated RAW264.7 cells stimulated with BTO and pBTO. (B) Relative mRNA levels of genes associated with osteoclastogenesis of RAW264.7 cells in the presence of RANKL, BTO or pBTO. (C) JC-1 staining in RANKL-treated RAW264.7 cells stimulated with BTO and pBTO. (D) Schematic diagram of pBTO inhibit osteoclastogenesis of macrophage (∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗∗: p < 0.001).

Article Snippet: Based on our previous research [ ], RAW 264.7 cells were incubated with 100 μg/mL BTO and pBTO, and stimulated with 50 ng/mL RANKL (peprotech, USA) for 3–5 days and then subjected to TRAP staining (Servicebio, China).

Techniques: In Vitro, Staining

Journal: Materials Today Bio

Article Title: Self-powered intracellular nanogenerator attenuates inflammatory osteolysis through mitochondrial MRS2/Mg 2+ -mediated macrophage repolarization and osteoclastogenesis inhibition

doi: 10.1016/j.mtbio.2025.102050

Figure Lengend Snippet: pBTO improved inflammatory bone loss by modulating macrophage polarization and osteoclastogenesis in vivo. (A) Designed an mice inflammatory bone loss model. (B) Micro-CT scanning and 3D reconstruction of mice calvarial bone. pBTO inhibited LPS-induced inflammatory bone loss. (C) CD68 + Arg-1 + macrophages (arrow) in calvarial bone matrix under confocal microscope. PBTO increased the number of M2 macrophages. (D) CD68 + iNOS + macrophages (arrow) in calvarial bone matrix under confocal microscope. pBTO decreased the number of LPS-induced M1 macrophages. (E) HE staining of mice calvarial bone showed pBTO decreased LPS-induced inflammatory cell infiltration and bone resorption. (F) TRAP staining of mice calvarial bone showed pBTO decreased the number of osteoclasts.

Article Snippet: Based on our previous research [ ], RAW 264.7 cells were incubated with 100 μg/mL BTO and pBTO, and stimulated with 50 ng/mL RANKL (peprotech, USA) for 3–5 days and then subjected to TRAP staining (Servicebio, China).

Techniques: In Vivo, Micro-CT, Microscopy, Staining

Journal: bioRxiv

Article Title: Targeting adipocyte ESRRA promotes osteogenesis and vascular formation in adipocyte-rich bone marrow

doi: 10.1101/2023.08.14.552932

Figure Lengend Snippet: (A) Schematic diagram illustrating the experimental procedure for diet-induced obesity (DIO) mice model. 9-week-old Esrra fl/fl and Esrra AKO male mice were fed a high-fat diet (HFD) for 16 weeks. (B) Representative pictures and body weights of DIO mice. (C) Representative images and weight analysis of white adipose tissue (WAT) depots, including gonadal WAT (gWAT), inguinal WAT (iWAT), and mesentery WAT (mWAT). (D) H&E-stained gWAT sections (scale bar: 50 μm). The areas of adipocytes size are presented as graphs. (E-F) Plasma TG (E) and FFA (F) levels. (G) Oral glucose tolerance test (OGTT) was analyzed in Esrra fl/fl and Esrra AKO mice fed a HFD for 12 weeks. (H) Plasma LEPTIN levels. (I) Representative micro-CT images of the distal femoral trabecular bone. (J) Quantitative analysis of bone volume/tissue volume ratio (BV/TV), trabecular thickness (Tb. Th), trabecular number (Tb. N) and trabecular separation (Tb. Sp). (K) H&E staining of femur sections (scale bar: 100 μm). Yellow arrows indicate the bone marrow adipocytes. Osteoblast surface to bone surface ratio (Ob.S/BS) and osteoblast number to bone surface ratio (Ob.N/BS) are shown on the right panel. (L) Calcein double labeling of trabecular bone (scale bar: 20 μm). Mineral apposition rate (MAR) and bone formation rate (BFR/BS) were determined. White arrows indicate the distance between calcein double labeling. (M) TRAP staining of femur sections with quantitative analysis of Oc.S/BS and Oc.N/BS. TRAP-positive purple spots indicate multinucleated osteoclasts (scale bar: 100 μm). (N-O) Plasma P1NP (N) and CTX1 (O) levels. (P) PLIN1 positive marrow adipocytes (PLIN1 + , red) and SPP1 (green) immunofluorescence staining in femur sections (scale bar: 100 μm). The box in the upper showing the metaphysis region near growth plate is represented at higher magnification in the bottom (scale bar: 50 μm). The numbers and areas of adipocytes in the femur marrow per tissue area and the quantification of SPP1 fluorescence intensity were measured. Data are shown as mean ± SD (n = 6 mice). * P < 0.05 was considered statistically significant.

Article Snippet: The paraffin-embedded femurs were subjected to deparaffinization and rehydration procedures before being stained using hematoxylin and eosin (H&E) kit (Beyotime #C0107) or tartrate-resistant acid phosphatase (TRAP) staining kit (Solarbio #G1492).

Techniques: Staining, Micro-CT, Labeling, Immunofluorescence, Fluorescence

Journal: bioRxiv

Article Title: Targeting adipocyte ESRRA promotes osteogenesis and vascular formation in adipocyte-rich bone marrow

doi: 10.1101/2023.08.14.552932

Figure Lengend Snippet: (A) Schematic diagram illustrating the experimental procedure for ovariectomy (OVX)-induced osteoporosis mice model. 10-week-old Esrra fl/fl and Esrra AKO female mice underwent either sham or OVX operation for 8 weeks. (B) Representative images and weights of adipose depots. (C) Representative images and adipocytes size analysis from H&E-stained gWAT sections (scale bar: 50 μm). (D) Plasma LEPTIN levels. (E-F) Micro-CT images of distal femurs in sham and OVX mice (E) with morphometric analysis of BV/TV, Tb.N, Tb.Th, and Tb.Sp (F). (G) Representative TRAP-stained images and quantification of Oc.S/BS and Oc.N/BS in distal femoral metaphysis regions from sham and OVX mice (scale bar: 100 μm). (H) Representative H&E-stained images and quantification of Ob.S/BS and Ob.N/BS (scale bar: 100 μm). (I-J) Plasma P1NP (I) and CTX1 (J) levels. (K) Calcein double labeling with quantitative analysis of MAR and BFR/BS (scale bar: 20 μm). (L) Immunofluorescence co-staining and quantification of PLIN1 + bone marrow adipocytes (red) and SPP1 (green) of femur sections from OVX mice. Scale bar: upper panel, 100 μm; lower panel, 50 μm. Data are shown as mean ± SD (n = 7 mice). * P <0.05 was considered statistically significant.

Article Snippet: The paraffin-embedded femurs were subjected to deparaffinization and rehydration procedures before being stained using hematoxylin and eosin (H&E) kit (Beyotime #C0107) or tartrate-resistant acid phosphatase (TRAP) staining kit (Solarbio #G1492).

Techniques: Staining, Micro-CT, Labeling, Immunofluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Periodontal Inflammation-Triggered by Periodontal Ligament Stem Cell Pyroptosis Exacerbates Periodontitis

doi: 10.3389/fcell.2021.663037

Figure Lengend Snippet: The paracrine effect of PDLSC pyroptosis on osteoblast differentiation and osteoclast differentiation. (A) Schematic diagram of coculture design indicating the placement of PDLSCs with LPS, with LPS FuGene treatment, or without treatment on the transwell insert with the lower chamber containing healthy PDLSCs. (B–H) Cells in the cocultured transwell system were maintained in osteogenic differentiation medium for 21 days. PDLSCs in a single culture (without coculture) were maintained in MEM-α medium as a control. (B) Entire plate views and micrographs of alizarin red staining after culturing in osteogenic differentiation medium. (C) Quantification of the alizarin red staining results. (D) Immunoblots for RUNX2, ALP, and OPN in whole-cell lysates. GAPDH was used as a loading control. RUNX2 (E) , ALP (F) , OPN (G) , and OSX (H) mRNAs were subjected to real-time PCR analysis, and the expression levels were normalized to that of 36B4 . (I) Schematic diagram of coculture design indicating the placement of PDLSCs with LPS treatment, with LPS FuGene treatment, or without treatment on transwell insert with the lower chamber containing THP-1 cells. (J–O) The THP-1 cells in the coculture system were cultured in osteoclast differentiation medium for 14 days. THP-1 cells in a single culture were maintained in RPMI-1640 medium as a control. (J) Micrographs of TRAP staining after culturing in osteoclast differentiation medium. (K) Quantification of TRAP-positive multinucleated cells (nuclei > 3). TRAP (L) , CTSK (M) , DCSTAMP (N) , and NFATC1 (O) mRNAs were subjected to real-time PCR analysis, and the expression levels were normalized to that of 36B4 . The data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01.

Article Snippet: After 14 days of induction, cells were fixed with 4% PFA and stained with TRAP staining solution (Jiancheng Biotech, China).

Techniques: Control, Staining, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Cell Culture

Journal: Frontiers in Cell and Developmental Biology

Article Title: Periodontal Inflammation-Triggered by Periodontal Ligament Stem Cell Pyroptosis Exacerbates Periodontitis

doi: 10.3389/fcell.2021.663037

Figure Lengend Snippet: IL-1β was the key factor in PDLSC pyroptosis to regulate osteoblast and osteoclast differentiation. (A) Expression profiles of several inflammatory cytokines in PDLSCs under LPS FuGene treatment for 16 h. (B–G) PDLSCs were incubated with osteogenic differentiation medium for 21 days with or without 500 pg/mL IL-1β treatment. (B) Representative images of alizarin red staining and quantification of the staining results. ALP (C) , OSX (D) , RUNX2 (E) , and OPN (F) mRNAs were subjected to real-time PCR analysis, and the expression levels were normalized to that of 36B4 . (G) Immunoblots for RUNX2, ALP, and OPN in whole-cell lysates. GAPDH was used as a loading control. (H–M) THP-1 cells were incubated with osteoclast differentiation medium for 14 days with or without 500 pg/mL IL-1β treatment. (H) Micrographs of TRAP staining after culturing in osteoclast differentiation medium. (I) Quantification of TRAP-positive multinucleated cells (nuclei > 3). TRAP (J) , CTSK (K) , DCSTAMP (L) , and NFATC1 (M) mRNAs were subjected to real-time PCR analysis, and the expression levels were normalized to that of 36B4 . The data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01.

Article Snippet: After 14 days of induction, cells were fixed with 4% PFA and stained with TRAP staining solution (Jiancheng Biotech, China).

Techniques: Expressing, Incubation, Staining, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Periodontal Inflammation-Triggered by Periodontal Ligament Stem Cell Pyroptosis Exacerbates Periodontitis

doi: 10.3389/fcell.2021.663037

Figure Lengend Snippet: Gsdmd –/– mice presented with reduced periodontal inflammation and bone loss in experimental periodontitis. (A–H) Mice were divided into the WT-sham group, WT-ligature group, and Gsdmd-KO-ligature group. Silk thread was used to form a ligature around the left maxillary first molar for 2 weeks to induce periodontitis. (A) Periodontium tissues were collected from each group, and the mRNA expression of osteoblast markers (including Runx2 , Alp , Opn , and Osx ) and osteoclast markers (including Trap , Ctsk , Dcstamp , and Nfact1 ) was analyzed by real-time PCR. 36b4 was used as an internal control. (B) Tissue sections from mice were prepared after 14 days of periodontitis induction and processed for hematoxylin and eosin (H&E) staining. (C) Representative micro-CT sagittal images of maxillary molars after insertion of ligature. CEJ, cement-enamel junction; ABC, alveolar bone crest. The line indicates the distance from the CEJ to the ABC on the buccal side of the ligature site. (D) The distances (μm) between CEJ and ABC were measured after periodontitis induction. (E) The images of bone surrounding first molars were analyzed by 3D micro-computed tomography. The squares formed by the continuous dotted line show visual differences in alveolar bone from animal groups. The bone mineral density (F) and bone/tissue volume (G) of the selected square regions on the ligature site were calculated. (H) Immunofluorescence staining of CD90 (green) in mouse maxillary molar longitudinal sections from the periodontitis model and sham group. Nuclei were identified by staining with DAPI. Scale bars, 50 μm. The data are presented as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: After 14 days of induction, cells were fixed with 4% PFA and stained with TRAP staining solution (Jiancheng Biotech, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Staining, Micro-CT, Immunofluorescence