Journal: Materials Today Bio

Article Title: Cationic microbubble loading hSIRT3 and hTIMP3 optimize cardiac-targeted delivery and myocardial protection in the porcine MI/R model

doi: 10.1016/j.mtbio.2025.102234

Figure Lengend Snippet: Characteristics of myocardial and cardiac function in a porcine MI/R injury model. a. Dynamic ECG in the establishment of porcine MI/R injury model. b. the level of LVEF, LVFS at baseline and days 3, 7, 14 after MI/R injury. c. Representative images for CD11b and Col1a1 expressions in IZ myocardium of the Sham group and on days 3, 7, and 14 after MI/R injury (scale bar = 100 μm). d. The protein expressions and quantitative analysis of endogenous SIRT3 and TIMP3 in IZ, BZ, and RZ in the Sham group and on days 3, 7, and 14 after MI/R injury (∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. Sham group). Data are presented as the Means ± SD (n = 3). ECG: Electrocardiogram. LV-EF: Left ventricular ejection fraction. LV-FS: Left ventricular fractional shortening. IZ: Infarction zone. BZ: Border zone. RZ: Remote zone.

Article Snippet: The primary antibodies against hSIRT3 (1:1,000, ab217319, Abcam), hTIMP3 (1:1,000, ab276134, Abcam), SIRT3 (1:1,000, 10099-1-AP, Proteintech), TIMP3 (1:1,000, 10858-1-AP, Proteintech), MnSOD (1:5,000, 24127-1-AP, Proteintech), CAT (1:2,000, 21260-1-AP, Proteintech), MMP2 (1:500, 10373-2-AP, Proteintech), MMP9 (1:1,000, 10375-2-AP, Proteintech), caspase-3 (1:1,000, 66470-2-Ig, Proteintech), Bcl2 (1:1,000, ab32124, Abcam) and Bax (1:5,000, 50599-2-Ig, Proteintech) were incubated with the corresponding PVDF membranes overnight at 4 °C.

Techniques:

Journal: Materials Today Bio

Article Title: Cationic microbubble loading hSIRT3 and hTIMP3 optimize cardiac-targeted delivery and myocardial protection in the porcine MI/R model

doi: 10.1016/j.mtbio.2025.102234

Figure Lengend Snippet: Characteristics of CMBs. a. Schematic diagram of CMB preparation. b. The appearance of CMBs. c. Microscopic morphology of CMBs detected by TEM (scale bar = 100 nm). d. The distributions of diameter and (e) surface charge of CMBs. f. The morphology of CMBs/vector labeled with PI stain (red). Scale bar = 50 μm. g. The encapsulation efficiency of CMBs for different doses of hSIRT3 or hTIMP3 genes. h. The loading rate of CMBs with hSIRT3 and hTIMP3 plasmids detected by flow cytometry. Data are presented as the Means ± SD (n = 3). CMBs: Cationic microbubbles. TEM: Transmission electron microscopy. hSIRT3: human SIRT3. hTIMP3: human TIMP3. ∗∗∗ P < 0.001. n.s.: not significant. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The primary antibodies against hSIRT3 (1:1,000, ab217319, Abcam), hTIMP3 (1:1,000, ab276134, Abcam), SIRT3 (1:1,000, 10099-1-AP, Proteintech), TIMP3 (1:1,000, 10858-1-AP, Proteintech), MnSOD (1:5,000, 24127-1-AP, Proteintech), CAT (1:2,000, 21260-1-AP, Proteintech), MMP2 (1:500, 10373-2-AP, Proteintech), MMP9 (1:1,000, 10375-2-AP, Proteintech), caspase-3 (1:1,000, 66470-2-Ig, Proteintech), Bcl2 (1:1,000, ab32124, Abcam) and Bax (1:5,000, 50599-2-Ig, Proteintech) were incubated with the corresponding PVDF membranes overnight at 4 °C.

Techniques: Plasmid Preparation, Labeling, Staining, Encapsulation, Flow Cytometry, Transmission Assay, Electron Microscopy

Journal: Materials Today Bio

Article Title: Cationic microbubble loading hSIRT3 and hTIMP3 optimize cardiac-targeted delivery and myocardial protection in the porcine MI/R model

doi: 10.1016/j.mtbio.2025.102234

Figure Lengend Snippet: Cardiac-targeted delivery of hSIRT3 and hITMP3 plasmids by UTMD. a. Schematic diagram of UTMD procedure for cardiac-targeted delivery of hSIRT3 and hITMP3 plasmids. b. Representative ultrasound contrast images of imaging and destructing CMBs/hSIRT3 or CMBs/hTIMP3 in the heart. c. Representative images of GFP expression in the myocardium and quantitative analysis of GPF expression among the groups. d. Protein expressions and quantitative analysis of hSIRT3 and hTIMP3 among the groups. e. Protein expressions and quantitative analysis of hSIRT3 and hTIMP3 among RZ, BZ and IZ myocardium. Data are presented as the Means ± SD (n = 3). UTMD: Ultrasound-targeted microbubble destruction. CMBs: Cationic microbubbles. hSIRT3: human SIRT3. hTIMP3: human TIMP3. IZ: Infarction zone. BZ: Border zone. RZ: Remote zone. ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: The primary antibodies against hSIRT3 (1:1,000, ab217319, Abcam), hTIMP3 (1:1,000, ab276134, Abcam), SIRT3 (1:1,000, 10099-1-AP, Proteintech), TIMP3 (1:1,000, 10858-1-AP, Proteintech), MnSOD (1:5,000, 24127-1-AP, Proteintech), CAT (1:2,000, 21260-1-AP, Proteintech), MMP2 (1:500, 10373-2-AP, Proteintech), MMP9 (1:1,000, 10375-2-AP, Proteintech), caspase-3 (1:1,000, 66470-2-Ig, Proteintech), Bcl2 (1:1,000, ab32124, Abcam) and Bax (1:5,000, 50599-2-Ig, Proteintech) were incubated with the corresponding PVDF membranes overnight at 4 °C.

Techniques: Imaging, Expressing

Journal: Materials Today Bio

Article Title: Cationic microbubble loading hSIRT3 and hTIMP3 optimize cardiac-targeted delivery and myocardial protection in the porcine MI/R model

doi: 10.1016/j.mtbio.2025.102234

Figure Lengend Snippet: Exogenous hSIRT3 and hTIMP3 genes exerted their downstream signaling regulation and inhibited the infiltration of inflammatory cells. a. Flowchart of short-term effects of exogenous hSIRT3 and hTIMP3 genes on myocardial injury. b. Protein expressions and quantitative analysis of CAT and MnSOD among the different groups. c. Protein expressions and quantitative analysis of MMP2 and MMP9 among the different groups. d. Representative images of CD11b expression in the myocardium and quantitative analysis of CD11b expression among the different groups (scale bar = 100 μm). Data are presented as the Means ± SD (n = 3). hSIRT3: human SIRT3. hTIMP3: human TIMP3. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. n.s.: not significant.

Article Snippet: The primary antibodies against hSIRT3 (1:1,000, ab217319, Abcam), hTIMP3 (1:1,000, ab276134, Abcam), SIRT3 (1:1,000, 10099-1-AP, Proteintech), TIMP3 (1:1,000, 10858-1-AP, Proteintech), MnSOD (1:5,000, 24127-1-AP, Proteintech), CAT (1:2,000, 21260-1-AP, Proteintech), MMP2 (1:500, 10373-2-AP, Proteintech), MMP9 (1:1,000, 10375-2-AP, Proteintech), caspase-3 (1:1,000, 66470-2-Ig, Proteintech), Bcl2 (1:1,000, ab32124, Abcam) and Bax (1:5,000, 50599-2-Ig, Proteintech) were incubated with the corresponding PVDF membranes overnight at 4 °C.

Techniques: Expressing

Journal: Materials Today Bio

Article Title: Cationic microbubble loading hSIRT3 and hTIMP3 optimize cardiac-targeted delivery and myocardial protection in the porcine MI/R model

doi: 10.1016/j.mtbio.2025.102234

Figure Lengend Snippet: Exogenous hSIRT3 and hTIMP3 genes inhibited apoptosis of myocardium. a. Representative images of IL-1β, IL-6 and TNF-α expressions in IZ myocardium and quantitative analysis (b) among the different groups (scale bar = 100 μm). c. Protein expressions and quantitative analysis of caspase-3, Bcl2, and Bax among the different groups. d. Representative images of TUNEL-positive cells in the myocardium and quantitative analysis of TUNEL-positive cells among the different groups (scale bar = 100 μm). Data are presented as the Means ± SD (n = 3). hSIRT3: human SIRT3. hTIMP3: human TIMP3. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. n.s.: not significant.

Article Snippet: The primary antibodies against hSIRT3 (1:1,000, ab217319, Abcam), hTIMP3 (1:1,000, ab276134, Abcam), SIRT3 (1:1,000, 10099-1-AP, Proteintech), TIMP3 (1:1,000, 10858-1-AP, Proteintech), MnSOD (1:5,000, 24127-1-AP, Proteintech), CAT (1:2,000, 21260-1-AP, Proteintech), MMP2 (1:500, 10373-2-AP, Proteintech), MMP9 (1:1,000, 10375-2-AP, Proteintech), caspase-3 (1:1,000, 66470-2-Ig, Proteintech), Bcl2 (1:1,000, ab32124, Abcam) and Bax (1:5,000, 50599-2-Ig, Proteintech) were incubated with the corresponding PVDF membranes overnight at 4 °C.

Techniques: TUNEL Assay

Journal: Materials Today Bio

Article Title: Cationic microbubble loading hSIRT3 and hTIMP3 optimize cardiac-targeted delivery and myocardial protection in the porcine MI/R model

doi: 10.1016/j.mtbio.2025.102234

Figure Lengend Snippet: Exogenous hSIRT3 and hTIMP3 genes promoted revascularization after myocardial injury. a. Flowchart of long-term effects of exogenous hSIRT3 and hTIMP3 genes on myocardial injury. b. Representative images of α-SMA labeled arteries and (c) CD31 labeled veins in the myocardium and quantitative analysis among the different groups (scale bar = 100 μm). Data are presented as the Means ± SD (n = 3). hSIRT3: human SIRT3. hTIMP3: human TIMP3. ∗∗ P < 0.01, ∗∗∗ P < 0.001. n.s.: not significant.

Article Snippet: The primary antibodies against hSIRT3 (1:1,000, ab217319, Abcam), hTIMP3 (1:1,000, ab276134, Abcam), SIRT3 (1:1,000, 10099-1-AP, Proteintech), TIMP3 (1:1,000, 10858-1-AP, Proteintech), MnSOD (1:5,000, 24127-1-AP, Proteintech), CAT (1:2,000, 21260-1-AP, Proteintech), MMP2 (1:500, 10373-2-AP, Proteintech), MMP9 (1:1,000, 10375-2-AP, Proteintech), caspase-3 (1:1,000, 66470-2-Ig, Proteintech), Bcl2 (1:1,000, ab32124, Abcam) and Bax (1:5,000, 50599-2-Ig, Proteintech) were incubated with the corresponding PVDF membranes overnight at 4 °C.

Techniques: Labeling

Journal: Materials Today Bio

Article Title: Cationic microbubble loading hSIRT3 and hTIMP3 optimize cardiac-targeted delivery and myocardial protection in the porcine MI/R model

doi: 10.1016/j.mtbio.2025.102234

Figure Lengend Snippet: Exogenous hSIRT3 and hTIMP3 genes suppressed myocardial fibrosis after myocardial injury. a. Representative images of collagen deposition and myocardial fibrosis of IZ stained by Masson's and SR assays and quantitative analysis among the different groups (scale bar = 100 μm). b. Dynamic values of LV-EF, LV-FS at baseline and different time points after gene therapy among the groups. c. Representative images and quantitative analysis of fibrotic scar among the different groups. Data are presented as the Means ± SD (n = 3). hSIRT3: human SIRT3. hTIMP3: human TIMP3. IZ: Infarction zone. BZ: Border zone. RZ: Remote zone. LV-EF: Left ventricular ejection fraction. LV-FS: Left ventricular fractional shortening. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. n.s.: not significant.

Article Snippet: The primary antibodies against hSIRT3 (1:1,000, ab217319, Abcam), hTIMP3 (1:1,000, ab276134, Abcam), SIRT3 (1:1,000, 10099-1-AP, Proteintech), TIMP3 (1:1,000, 10858-1-AP, Proteintech), MnSOD (1:5,000, 24127-1-AP, Proteintech), CAT (1:2,000, 21260-1-AP, Proteintech), MMP2 (1:500, 10373-2-AP, Proteintech), MMP9 (1:1,000, 10375-2-AP, Proteintech), caspase-3 (1:1,000, 66470-2-Ig, Proteintech), Bcl2 (1:1,000, ab32124, Abcam) and Bax (1:5,000, 50599-2-Ig, Proteintech) were incubated with the corresponding PVDF membranes overnight at 4 °C.

Techniques: Staining