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In vitro evaluation of osteogenic differentiation on Mg 2+ -releasing piezoelectric scaffolds. A) <t>ALP</t> <t>staining</t> of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). B) ARS staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). C-F) RT-qPCR results showing the relative mRNA expression of OPN, RUNX2, OCN, <t>and</t> <t>COL-I</t> in BMSCs cultured with cryogels for 7 days and 14 days. CLSM images showing the expression of (G) OPN, (H) RUNX2, (I) OCN, and (J) COL-I in BMSCs co-cultured with WH Gel and PWH Gel (Scale bar: 50 μm). Data are presented as mean ± S.D. (n = 3 independent replicates). ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Col I, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 261 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair"

Article Title: Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair

Journal: Bioactive Materials

doi: 10.1016/j.bioactmat.2026.02.017

In vitro evaluation of osteogenic differentiation on Mg 2+ -releasing piezoelectric scaffolds. A) ALP staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). B) ARS staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). C-F) RT-qPCR results showing the relative mRNA expression of OPN, RUNX2, OCN, and COL-I in BMSCs cultured with cryogels for 7 days and 14 days. CLSM images showing the expression of (G) OPN, (H) RUNX2, (I) OCN, and (J) COL-I in BMSCs co-cultured with WH Gel and PWH Gel (Scale bar: 50 μm). Data are presented as mean ± S.D. (n = 3 independent replicates). ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Figure Legend Snippet: In vitro evaluation of osteogenic differentiation on Mg 2+ -releasing piezoelectric scaffolds. A) ALP staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). B) ARS staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). C-F) RT-qPCR results showing the relative mRNA expression of OPN, RUNX2, OCN, and COL-I in BMSCs cultured with cryogels for 7 days and 14 days. CLSM images showing the expression of (G) OPN, (H) RUNX2, (I) OCN, and (J) COL-I in BMSCs co-cultured with WH Gel and PWH Gel (Scale bar: 50 μm). Data are presented as mean ± S.D. (n = 3 independent replicates). ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Techniques Used: In Vitro, Staining, Cell Culture, Quantitative RT-PCR, Expressing

In vivo assessments of large segmental bone defect regeneration using Mg 2+ -releasing piezoelectric scaffold. A-B) Schematic showing the surgical procedure for scaffold implantation in rat radial defects (Scale bar = 1 cm). C) Macroscopic images of the defect site at 6- and 12- weeks post-implantation. D) RUS scores for radial repair. E) 3D micro-CT images of the defects at 6- and 12- weeks post-implantation (Scale bar = 3 mm). F-G) Quantitative micro-CT analysis of BV/TV and trabecular number (Tb.N) in cryogel-treated regions at 6- and 12- weeks post-implantation. H) Representative H&E and Masson's trichrome staining images of defect tissues at 6- and 12-weeks post-implantation (Scale bar: 1 mm). I) Immunohistochemical staining for COL-I (Scale bar: 1 mm). J) Representative immunofluorescence staining of CD31 (Scale bar: 1 mm). Data are expressed as mean ± S.D. (n = 3 independent replicates). Statistical significance was determined as ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Figure Legend Snippet: In vivo assessments of large segmental bone defect regeneration using Mg 2+ -releasing piezoelectric scaffold. A-B) Schematic showing the surgical procedure for scaffold implantation in rat radial defects (Scale bar = 1 cm). C) Macroscopic images of the defect site at 6- and 12- weeks post-implantation. D) RUS scores for radial repair. E) 3D micro-CT images of the defects at 6- and 12- weeks post-implantation (Scale bar = 3 mm). F-G) Quantitative micro-CT analysis of BV/TV and trabecular number (Tb.N) in cryogel-treated regions at 6- and 12- weeks post-implantation. H) Representative H&E and Masson's trichrome staining images of defect tissues at 6- and 12-weeks post-implantation (Scale bar: 1 mm). I) Immunohistochemical staining for COL-I (Scale bar: 1 mm). J) Representative immunofluorescence staining of CD31 (Scale bar: 1 mm). Data are expressed as mean ± S.D. (n = 3 independent replicates). Statistical significance was determined as ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Techniques Used: In Vivo, Micro-CT, Staining, Immunohistochemical staining, Immunofluorescence

Immunohistochemical staining:

Article Title: Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair.
Article Snippet: Histological staining was performed following the manufacturer's guidelines, using Hematoxylin and Eosin (H&E) staining kit (G1121, Solarbio, China) and Masson's Trichrome Staining Kit (G1340, Solarbio, China). .. Immunohistochemical staining was carried out for COL-I (Proteintech, 28083-1-AP, USA). .. Immunofluorescence staining was performed using primary antibodies against CD31 (Abcam, ab34710, USA) and Tuj1 (Proteintech, 66375-1-Ig, USA).

Article Title: Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair
Article Snippet: .. Immunohistochemical staining was carried out for COL-I (Proteintech, 28083-1-AP, USA). .. Immunofluorescence staining was performed using primary antibodies against CD31 (Abcam, ab34710, USA) and Tuj1 (Proteintech, 66375-1-Ig, USA).

Staining:

Electrophoresis:

Article Title: Huanggan decoction ameliorates cholestatic hepatic fibrosis in rats via TGF-β1/Smad3 signaling pathway
Article Snippet: .. Following electrophoresis and membrane transfer, the blots were blocked by skim milk and then incubated with specific antibodies to MMP1 (26585–1-AP, Proteintech, Wuhan), TIMP1 (16644–1-AP, Proteintech, Wuhan), Smad3 (66516–1-Ig, Proteintech, Wuhan), p-Smad3(80427–2-RR, Proteintech, Wuhan), Col-I (67288–1-Ig, Proteintech, Wuhan), α-SMA (14395–1-AP, Proteintech, Wuhan) and GAPDH (60004–1-Ig, Proteintech, Wuhan) overnight at 4°C. .. Following a 1-hour incubation at ambient temperature with the secondary antibodies (KFA021, Proteintech, Wuhan), the signals were identified and the protein band intensities were measured using Qinxiang chemical analysis software from Clinx Science Instruments, Shanghai.

Article Title: Huanggan decoction ameliorates cholestatic hepatic fibrosis in rats via TGF-β1/Smad3 signaling pathway.
Article Snippet: .. Following electrophoresis and membrane transfer, the blots were blocked by skim milk and then incubated with specific antibodies to MMP1 (26585–1-AP, Proteintech, Wuhan), TIMP1 (16644–1-AP, Proteintech, Wuhan), Smad3 (66516–1-Ig, Proteintech, Wuhan), p-Smad3(80427–2-RR, Proteintech, Wuhan), Col-I (67288–1-Ig, Proteintech, Wuhan), α-SMA (14395–1-AP, Proteintech, Wuhan) and GAPDH (60004–1-Ig, Proteintech, Wuhan) overnight at 4°C. .. Following a 1-hour incubation at ambient temperature with the secondary antibodies (KFA021, Proteintech, Wuhan), the signals were identified and the protein band intensities were measured using Qinxiang chemical analysis software from Clinx Science Instruments, Shanghai.

Membrane:

Article Title: 3D printing combined with thermally induced phase separation for engineering hierarchical osteogenic PLA scaffolds
Article Snippet: .. The membrane was blocked with 5 % skim milk for 1 h and incubated with primary antibodies against Runx2 (1:500, Proteintech, China), Ocn (1:500, Abcam, UK), Opn (1:500, Proteintech, China), Col-I (1:1000, Proteintech, China), anti-Beta actin (1:20000, Proteintech, USA), and anti-Macf1 (1:1000, Abcam, UK) at 4 °C overnight. .. Horseradish peroxidase (HRP)-conjugated secondary antibodies (Pierce, Rockford, USA) were used, and protein bands were visualized using an ECL kit (Pioneer Biotechnology, China) and exposed to a chemiluminescence analyzer.

Incubation:

Article Title: An Electrospun Heparin Modified Nanofiber Membrane Carrying Multiple Growth Factors for Dental Pulp Regeneration
Article Snippet: Then the proteins were transferred onto polyvinylidene difluoride membranes (PVDF, Millipore, MA, USA). .. The membranes were blocked with 5% skim milk and then incubated at 4°C overnight with the following primary antibodies: VEGFR2 (1:1000; Abclonal Technology, Wuhan, China), CD31 (1:5000; Proteintech Group), COL-I (1:1000; Proteintech Group), DMP1 (1:1000; Abclonal, Wuhan, China), TUBB3 (1:5000; Abmart, Shanghai, China), NEUN (1:500; ZENBIO, Chengdu, China), and GAPDH (1:5000; Proteintech, Chicago, USA). .. After being incubated with secondary antibodies (Proteintech Group), the membranes were detected by ECL solution (Yeasen Biotechnology Co, Ltd).

Article Title: Early Intervention of Subchondral Bone Resorption Mitigates Cartilage Degeneration in TMJOA.
Article Snippet: Background: Excessive subchondral bone resorption is a typical manifestation in the early stage of osteoarthritis.. This study is to verify whether and when intervening in subchondral bone could alleviate cartilage degeneration of temporomandibular joint

Blocking Assay:

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Journal: Bioactive Materials

Article Title: Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair

doi: 10.1016/j.bioactmat.2026.02.017

Figure Lengend Snippet: In vitro evaluation of osteogenic differentiation on Mg 2+ -releasing piezoelectric scaffolds. A) ALP staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). B) ARS staining of BMSCs cultured with WH Gel and PWH Gel (Scale bar: 1 mm). C-F) RT-qPCR results showing the relative mRNA expression of OPN, RUNX2, OCN, and COL-I in BMSCs cultured with cryogels for 7 days and 14 days. CLSM images showing the expression of (G) OPN, (H) RUNX2, (I) OCN, and (J) COL-I in BMSCs co-cultured with WH Gel and PWH Gel (Scale bar: 50 μm). Data are presented as mean ± S.D. (n = 3 independent replicates). ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Article Snippet: Immunohistochemical staining was carried out for COL-I (Proteintech, 28083-1-AP, USA).

Techniques: In Vitro, Staining, Cell Culture, Quantitative RT-PCR, Expressing

Journal: Bioactive Materials

Article Title: Biodegradable Mg 2+ -releasing piezoelectric scaffold for segmental bone defect repair

doi: 10.1016/j.bioactmat.2026.02.017

Figure Lengend Snippet: In vivo assessments of large segmental bone defect regeneration using Mg 2+ -releasing piezoelectric scaffold. A-B) Schematic showing the surgical procedure for scaffold implantation in rat radial defects (Scale bar = 1 cm). C) Macroscopic images of the defect site at 6- and 12- weeks post-implantation. D) RUS scores for radial repair. E) 3D micro-CT images of the defects at 6- and 12- weeks post-implantation (Scale bar = 3 mm). F-G) Quantitative micro-CT analysis of BV/TV and trabecular number (Tb.N) in cryogel-treated regions at 6- and 12- weeks post-implantation. H) Representative H&E and Masson's trichrome staining images of defect tissues at 6- and 12-weeks post-implantation (Scale bar: 1 mm). I) Immunohistochemical staining for COL-I (Scale bar: 1 mm). J) Representative immunofluorescence staining of CD31 (Scale bar: 1 mm). Data are expressed as mean ± S.D. (n = 3 independent replicates). Statistical significance was determined as ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; NS, not significant.

Article Snippet: Immunohistochemical staining was carried out for COL-I (Proteintech, 28083-1-AP, USA).

Techniques: In Vivo, Micro-CT, Staining, Immunohistochemical staining, Immunofluorescence

Involvement of profibrotic macrophages in vascular regeneration after graft implantation in vivo . (a) UMAP of macrophages in native aortas and regenerated aortas across different time points after graft implantation in vivo . (b) Dot plots of profibrotic macrophage marker genes (Ctsd, Spp1, Gpnmb, Lgals3, and Fabp5) expressed in different subgroups of macrophages. (c) Percentage of cluster 2 (C2) macrophages in native aortas and regenerated aortas across different time points after graft implantation in vivo . (d) UMAP of expression of Ctsd, Spp1, Gpnmb, Lgals3, and Fabp5 in macrophages in native aortas and regenerated aortas across different time points after graft implantation in vivo . (e) Immunofluorescence staining of CD68 and CTSD in regenerated aortas across different time points after graft implantation in vivo . L indicates lumens. Arrow heads indicate double positively stained cells. (f) WB results of levels of CTSD and SPP1 in native and regenerated aortas across different time points after graft implantation in vivo and quantification of the levels of CTSD and SPP1. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each time point and each group, five different samples from five different animals were analyzed (n = 5).

Journal: Bioactive Materials

Article Title: Apolipoprotein E knockout attenuates vascular graft fibrosis by reducing profibrotic macrophage formation through low-density lipoprotein receptor related protein 1

doi: 10.1016/j.bioactmat.2026.01.029

Figure Lengend Snippet: Involvement of profibrotic macrophages in vascular regeneration after graft implantation in vivo . (a) UMAP of macrophages in native aortas and regenerated aortas across different time points after graft implantation in vivo . (b) Dot plots of profibrotic macrophage marker genes (Ctsd, Spp1, Gpnmb, Lgals3, and Fabp5) expressed in different subgroups of macrophages. (c) Percentage of cluster 2 (C2) macrophages in native aortas and regenerated aortas across different time points after graft implantation in vivo . (d) UMAP of expression of Ctsd, Spp1, Gpnmb, Lgals3, and Fabp5 in macrophages in native aortas and regenerated aortas across different time points after graft implantation in vivo . (e) Immunofluorescence staining of CD68 and CTSD in regenerated aortas across different time points after graft implantation in vivo . L indicates lumens. Arrow heads indicate double positively stained cells. (f) WB results of levels of CTSD and SPP1 in native and regenerated aortas across different time points after graft implantation in vivo and quantification of the levels of CTSD and SPP1. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each time point and each group, five different samples from five different animals were analyzed (n = 5).

Article Snippet: The following primary antibodies were used in this study: APOE (Abcam, ab183597, 1:1000 dilution), COL I (abcam, ab270993, 1:1000 dilution), FN (abcam, ab268020, 1:1000 dilution), CTSD (CST, 74089S, 1:1000 dilution), SPP1 (NeoBiotechnologies, 6696-RBM3-P0, 1:1000 dilution), and LRP1 (Invitrogen, PA5-101013, 1:1000 dilution).

Techniques: In Vivo, Marker, Expressing, Immunofluorescence, Staining

APOE KO reducing profibrotic macrophage formation during vascular regeneration. (a) UMAP of macrophages in native aortas from WT and Apoe −/− rats, heatmap of C2 scores in the UMAP of macrophages in the native aortas, and percentage of C2 cells in macrophages in the native aortas. UMAP of macrophages in regenerated aortas after graft implantation in WT and Apoe −/− rats, heatmap of C2 scores in the UMAP of macrophages in the regenerated aortas, and percentage of C2 cells in macrophages in the regenerated aortas on Day 30 (b) and Day 90 (c). (d) Immunofluorescence staining of CD68 and CTSD in regenerated aortas 30 and 90 days after graft implantation in WT and Apoe−/− rats. (e) Quantification of CD68 and CTSD double positive cells in regenerated aortas on Day 30 and Day 90. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each time point and each group, five different images from five different samples were analyzed (n = 5). (f) WB results of APOE, CTSD and SPP1 levels in regenerated aortas after graft implantation in WT and Apoe −/− rats for 30 and 90 days. (g) Quantification of levels of APOE, CTSD and SPP1 in regenerated aortas on Day 30 and Day 90. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each time point and each group, five different samples from five different animals were analyzed (n = 5). (h) WB results of APOE, CTSD and SPP1 levels in WT and APOE KO macrophages after their culture on PCL scaffolds for 48 h. (i) Quantification of levels of APOE, CTSD and SPP1 in WT and APOE KO macrophages after their culture on PCL scaffolds for 48 h ∗ indicates p < 0.05, ∗∗ indicates p < 0.01, unpaired t -test. For each time point and each group, three different samples were analyzed (n = 3). (j) Immunofluorescence staining of APOE and CD68, CTSD and CD68, SPP1 and CD68, respectively, in WT and APOE KO macrophages after their culture on PCL scaffolds for 48 h.

Journal: Bioactive Materials

Article Title: Apolipoprotein E knockout attenuates vascular graft fibrosis by reducing profibrotic macrophage formation through low-density lipoprotein receptor related protein 1

doi: 10.1016/j.bioactmat.2026.01.029

Figure Lengend Snippet: APOE KO reducing profibrotic macrophage formation during vascular regeneration. (a) UMAP of macrophages in native aortas from WT and Apoe −/− rats, heatmap of C2 scores in the UMAP of macrophages in the native aortas, and percentage of C2 cells in macrophages in the native aortas. UMAP of macrophages in regenerated aortas after graft implantation in WT and Apoe −/− rats, heatmap of C2 scores in the UMAP of macrophages in the regenerated aortas, and percentage of C2 cells in macrophages in the regenerated aortas on Day 30 (b) and Day 90 (c). (d) Immunofluorescence staining of CD68 and CTSD in regenerated aortas 30 and 90 days after graft implantation in WT and Apoe−/− rats. (e) Quantification of CD68 and CTSD double positive cells in regenerated aortas on Day 30 and Day 90. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each time point and each group, five different images from five different samples were analyzed (n = 5). (f) WB results of APOE, CTSD and SPP1 levels in regenerated aortas after graft implantation in WT and Apoe −/− rats for 30 and 90 days. (g) Quantification of levels of APOE, CTSD and SPP1 in regenerated aortas on Day 30 and Day 90. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each time point and each group, five different samples from five different animals were analyzed (n = 5). (h) WB results of APOE, CTSD and SPP1 levels in WT and APOE KO macrophages after their culture on PCL scaffolds for 48 h. (i) Quantification of levels of APOE, CTSD and SPP1 in WT and APOE KO macrophages after their culture on PCL scaffolds for 48 h ∗ indicates p < 0.05, ∗∗ indicates p < 0.01, unpaired t -test. For each time point and each group, three different samples were analyzed (n = 3). (j) Immunofluorescence staining of APOE and CD68, CTSD and CD68, SPP1 and CD68, respectively, in WT and APOE KO macrophages after their culture on PCL scaffolds for 48 h.

Techniques: Immunofluorescence, Staining

APOE/LRP1 interaction promoting profibrotic macrophage formation during vascular regeneration after graft implantation in vivo . (a) Immunoprecipitation (IP) following mass spectrometry (MS) to screen potential receptors of APOE on surfaces of macrophages. (b) Co-immunoprecipitation (Co-IP) to confirm interaction between APOE and LRP1. (c) Immunofluorescence staining of CD68 and LRP1 in regenerated aortas across different time points. (d) Immunofluorescence staining of APOE and LRP1 in WT macrophages 48 h after their culture on PCL scaffolds. (e) WB results of LRP1, APOE, CTSD and SPP1 levels in WT macrophages cultured on tissue culture plates (negative control, NC) or PCL scaffolds (PCL) for 48 h prior to treatment with shRNA ADV-shRNA(NC) or ADV-shRNA(Lrp1) for 24 h. Quantification of levels of LRP1 (f), APOE (g), CTSD (h) and SPP1 (i) in WT macrophages cultured on tissue culture plates or PCL scaffolds treated with shRNA ADV-shRNA(NC) or ADV-shRNA(Lrp1). ∗ indicates p < 0.05, ∗∗ indicates p < 0.01, N.S. indicates non-significant. Tukey's post-hoc test. For each group, three different samples were analyzed (n = 3). (j) Flow cytometry analysis of CTSD positive cells in WT macrophages cultured on tissue culture plates (negative control, NC) or PCL scaffolds (PCL) for 48 h prior to treatment with ADV-shRNA(NC) or ADV-shRNA(Lrp1) for 24 h and quantification of percentage of CTSD positive cells in WT macrophages in each group. ∗ indicates p < 0.05, Tukey's post-hoc test. For each group, three independent experiments were repeated, and results were analyzed (n = 3).

Journal: Bioactive Materials

Article Title: Apolipoprotein E knockout attenuates vascular graft fibrosis by reducing profibrotic macrophage formation through low-density lipoprotein receptor related protein 1

doi: 10.1016/j.bioactmat.2026.01.029

Figure Lengend Snippet: APOE/LRP1 interaction promoting profibrotic macrophage formation during vascular regeneration after graft implantation in vivo . (a) Immunoprecipitation (IP) following mass spectrometry (MS) to screen potential receptors of APOE on surfaces of macrophages. (b) Co-immunoprecipitation (Co-IP) to confirm interaction between APOE and LRP1. (c) Immunofluorescence staining of CD68 and LRP1 in regenerated aortas across different time points. (d) Immunofluorescence staining of APOE and LRP1 in WT macrophages 48 h after their culture on PCL scaffolds. (e) WB results of LRP1, APOE, CTSD and SPP1 levels in WT macrophages cultured on tissue culture plates (negative control, NC) or PCL scaffolds (PCL) for 48 h prior to treatment with shRNA ADV-shRNA(NC) or ADV-shRNA(Lrp1) for 24 h. Quantification of levels of LRP1 (f), APOE (g), CTSD (h) and SPP1 (i) in WT macrophages cultured on tissue culture plates or PCL scaffolds treated with shRNA ADV-shRNA(NC) or ADV-shRNA(Lrp1). ∗ indicates p < 0.05, ∗∗ indicates p < 0.01, N.S. indicates non-significant. Tukey's post-hoc test. For each group, three different samples were analyzed (n = 3). (j) Flow cytometry analysis of CTSD positive cells in WT macrophages cultured on tissue culture plates (negative control, NC) or PCL scaffolds (PCL) for 48 h prior to treatment with ADV-shRNA(NC) or ADV-shRNA(Lrp1) for 24 h and quantification of percentage of CTSD positive cells in WT macrophages in each group. ∗ indicates p < 0.05, Tukey's post-hoc test. For each group, three independent experiments were repeated, and results were analyzed (n = 3).

Techniques: In Vivo, Immunoprecipitation, Mass Spectrometry, Co-Immunoprecipitation Assay, Immunofluorescence, Staining, Cell Culture, Negative Control, shRNA, Flow Cytometry

Downregulation of APOE by AAV ameliorating fibrosis during vascular regeneration after graft implantation in vivo . (a) Illustration of a strategy of adventitial delivery of AAV-shRNA(Apoe) to inhibit APOE levels in regenerated aortas after graft implantation in vivo . Two weeks after graft implantation in vivo , AAV-shRNA(Apoe) were injected into the adventitia of the regenerated aortas, which were then harvested for analysis three weeks later. (b) M mode images of ultrasound detection of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. Arrow heads indicate movement of vascular walls. (c) Tensile tests of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. (d) Quantification of RI, PI, and compliance of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different images from six different animals were analyzed (n = 6). (e) Quantification of elastic modulus of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different images from six different animals were analyzed (n = 6). (f) H&E, MTC and EVG staining of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. (g) Immunofluorescence staining of COL I, COL III, elastin, αSMA, and eNOS in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. L indicates lumens. Arrow heads indicate capillaries. Quantification of adventitia thickness (h), collagen positive areas according to MTC staining (i), elastin positive areas according to EVG staining (j), COL I positive areas (k), COL III positive areas (l), and number of capillaries (m) in adventitial areas of regenerated aortas. (n) Immunofluorescence staining of CTSD and CD68 in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. (o) CD68 and CTSD double positive cells in regenerated aortas. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different samples from six different animals were analyzed (n = 6). (p) WB results of APOE, CTSD and SPP1 levels in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks and quantification of levels of APOE, CTSD and SPP1 in regenerated aortas. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different samples from six different animals were analyzed (n = 6). (q) Quantification of IGF-1 concentrations in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks by ELISA. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different samples from six different animals were analyzed (n = 3).

Journal: Bioactive Materials

Article Title: Apolipoprotein E knockout attenuates vascular graft fibrosis by reducing profibrotic macrophage formation through low-density lipoprotein receptor related protein 1

doi: 10.1016/j.bioactmat.2026.01.029

Figure Lengend Snippet: Downregulation of APOE by AAV ameliorating fibrosis during vascular regeneration after graft implantation in vivo . (a) Illustration of a strategy of adventitial delivery of AAV-shRNA(Apoe) to inhibit APOE levels in regenerated aortas after graft implantation in vivo . Two weeks after graft implantation in vivo , AAV-shRNA(Apoe) were injected into the adventitia of the regenerated aortas, which were then harvested for analysis three weeks later. (b) M mode images of ultrasound detection of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. Arrow heads indicate movement of vascular walls. (c) Tensile tests of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. (d) Quantification of RI, PI, and compliance of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different images from six different animals were analyzed (n = 6). (e) Quantification of elastic modulus of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different images from six different animals were analyzed (n = 6). (f) H&E, MTC and EVG staining of regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. (g) Immunofluorescence staining of COL I, COL III, elastin, αSMA, and eNOS in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. L indicates lumens. Arrow heads indicate capillaries. Quantification of adventitia thickness (h), collagen positive areas according to MTC staining (i), elastin positive areas according to EVG staining (j), COL I positive areas (k), COL III positive areas (l), and number of capillaries (m) in adventitial areas of regenerated aortas. (n) Immunofluorescence staining of CTSD and CD68 in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks. (o) CD68 and CTSD double positive cells in regenerated aortas. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different samples from six different animals were analyzed (n = 6). (p) WB results of APOE, CTSD and SPP1 levels in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks and quantification of levels of APOE, CTSD and SPP1 in regenerated aortas. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different samples from six different animals were analyzed (n = 6). (q) Quantification of IGF-1 concentrations in regenerated aortas treated with PBS, AAV-shRNA(NC), and AAV-shRNA(Apoe) for 3 weeks by ELISA. ∗∗ indicates p < 0.01, Tukey's post-hoc test. For each group, six different samples from six different animals were analyzed (n = 3).

Techniques: In Vivo, shRNA, Injection, Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay

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col i (Proteintech)

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