Journal: The World Journal of Men's Health

Article Title: MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4

doi: 10.5534/wjmh.230149

Figure Lengend Snippet: The construction of EdU + BMSCs and gene modified BMSCs. (A) Fluorescence images of EdU (green) labeling of BMSCs. (B) The quantification of EdU + cells. (C) The cell viability of the labeled BMSCs with EdU at different concentrations for 24 hours. (D) Fluorescence images of miR-145 gene modified BMSCs. (E) The qRT-PCR results of miR-145. The data is represented as the mean±standard deviation. EdU: 5-ethynyl-2′-deoxyuridine, BMSCs: bone marrow-derived mesenchymal stem cells, miR-145: microRNA-145, qRT-PCR: quantitative real-time polymerase chain reaction. * p<0.05 compared with the 20 µm/24 hours group and ** p<0.01 compared with the control group, scale=100 µm, n=3 per group.

Article Snippet: The lentivirus-miR-145-green fluorescent proteins (GFP) (Hanbio Biotechnology ) and primary BMSCs (Cyagen Biosciences) derived from rats were prepared.

Techniques: Modification, Fluorescence, Labeling, Quantitative RT-PCR, Standard Deviation, Derivative Assay, Real-time Polymerase Chain Reaction, Control

Journal: The World Journal of Men's Health

Article Title: MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4

doi: 10.5534/wjmh.230149

Figure Lengend Snippet: The apoptosis and migration of BMSCs were inhibited by miR-145. (A) The apoptosis results illustrated by flow cytometry. (B) The apoptotic rates were calculated. (C) The proliferation of BMSCs of each group was assessed by using CCK-8 assays. (D) The migration of BMSCs of each group was evaluated by transwell assays (crystal violet staining, ×200). (E) The cell migration results were quantified. The data is represented as the means±standard deviation. BMSCs: bone marrow-derived mesenchymal stem cells, miR-145: microRNA-145, CCK-8: cell counting kit-8. ** p<0.01 compared with the control group, scale=100 µm, n=3 per group.

Article Snippet: The lentivirus-miR-145-green fluorescent proteins (GFP) (Hanbio Biotechnology ) and primary BMSCs (Cyagen Biosciences) derived from rats were prepared.

Techniques: Migration, Flow Cytometry, CCK-8 Assay, Staining, Standard Deviation, Derivative Assay, Cell Counting, Control

Journal: The World Journal of Men's Health

Article Title: MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4

doi: 10.5534/wjmh.230149

Figure Lengend Snippet: The differentiation of BMSCs into SMLCs enhanced by miR-145. (A) Fluorescence images of phalloidin staining (green represents F-actin). (B) Quantitative analysis of phalloidin staining. (C) The qRT-PCR results of α-actin and desmin. (D, E) The western blot results of α-actin and desmin. (F, G) The immunofluorescence results of α-actin. (H, I) The immunofluorescence results of desmin. (J) Cell contraction in each group under the stimulation of carbachol (×200). (K) Quantitative analysis of cell surface areas in each group. The data is represented as the means±standard deviation. BMSCs: bone marrow-derived mesenchymal stem cells, SMLCs: smooth muscle-like cells, DAPI: 4′,6-diamidino-2-phenylindole, GAPDH: glyceraldehyde 3-phosphate dehydrogenase, qRT-PCR: quantitative real-time polymerase chain reaction. * p<0.05 and ** p<0.01 compared with the control group, scale=100 µm, n=3.

Article Snippet: The lentivirus-miR-145-green fluorescent proteins (GFP) (Hanbio Biotechnology ) and primary BMSCs (Cyagen Biosciences) derived from rats were prepared.

Techniques: Fluorescence, Staining, Quantitative RT-PCR, Western Blot, Immunofluorescence, Standard Deviation, Derivative Assay, Real-time Polymerase Chain Reaction, Control

Journal: The World Journal of Men's Health

Article Title: MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4

doi: 10.5534/wjmh.230149

Figure Lengend Snippet: The retention and differentiation of BMSCs within CC after different treatments. (A) Representative images of EdU + cells within CC after 7 days of different treatments. (B) Representative images of EdU + cells within CC after 14 days of different treatments. (C) Quantitative analysis of the amount of EdU positive cells within CC after 7 and 14 days of different treatments. (D) Representative images of EdU/α-actin co-staining cells within CC after 7 days and 14 days of the different treatments. (E) Representative images of EdU/CD31 co-staining cells within CC after 7 days and 14 days of different treatments. (F) Quantitative analysis of EdU/CD31 co-staining cell amount within CC. The EdU (red)/DAPI (blue) fluorescence co-staining represents the EdU + cells. The EdU (red)/CD31 (green)/DAPI (blue) fluorescence co-staining represents the positive cells. The data is represented as the means±standard deviation, n=3. EdU: 5-ethynyl-2′-deoxyuridine, BMSCs: bone marrow-derived mesenchymal stem cells, miR-145: microRNA-145, PBS: phosphate buffer saline, DAPI: 4′,6-diamidino-2-phenylindole, CD31: platelet endothelial cell adhesion molecule-1. * p<0.05 and ** p<0.01 compared with the BMSCs group at the same time point. ## p<0.01 compared with the same treatment group at the different time point, scale=100 µm.

Article Snippet: The lentivirus-miR-145-green fluorescent proteins (GFP) (Hanbio Biotechnology ) and primary BMSCs (Cyagen Biosciences) derived from rats were prepared.

Techniques: Staining, Fluorescence, Standard Deviation, Derivative Assay, Saline

Journal: The World Journal of Men's Health

Article Title: MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4

doi: 10.5534/wjmh.230149

Figure Lengend Snippet: The target and function of OE-miR-145-BMSC. (A) The qRT-PCR results of KLF4 after the different treatments. (B, C) The IHC results of KLF4 (immunohistochemical staining, ×200). (D) The migration and apoptosis of OE-miR-145-BMSC of each group were evaluated by transwell and TUNEL assays (crystal violet staining, ×200). (E, F) The quantitative analysis of the results of cell migration and apoptosis. (G, I) The TUNEL staining results of CCSMCs in each group. (H, J) The IF staining of α-actin of CCSMCs in each group. The data is represented as the means±standard deviation. KLF4: Krüppel-like factor 4, miR-145: microRNA-145, BMSCs: bone marrow-derived mesenchymal stem cells, PBS: phosphate buffer saline, CCSMCs: corpus cavernosum smooth muscle cells, IF: immunofluorescence, DAPI: 4′,6-diamidino-2-phenylindole. * p<0.05 and ** p<0.01 compared with the control group, scale=100 µm, n=3.

Article Snippet: The lentivirus-miR-145-green fluorescent proteins (GFP) (Hanbio Biotechnology ) and primary BMSCs (Cyagen Biosciences) derived from rats were prepared.

Techniques: Quantitative RT-PCR, Immunohistochemical staining, Staining, Migration, TUNEL Assay, Standard Deviation, Derivative Assay, Saline, Immunofluorescence, Control

Journal: The World Journal of Men's Health

Article Title: MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4

doi: 10.5534/wjmh.230149

Figure Lengend Snippet: Schematic diagram illustrates the mechanism by which miR-145 gene modification modulates the traits and fate of BMSCs. In vitro , miR-145 inhibited the apoptosis and migration of BMSCs and promoted the differentiation of BMSCs into SMLCs. In vivo , after the ICI of OE-miR-145-BMSC, the prolonged retention of BMSCs and endothelioid cells were detected. However, no evidence supported that miR-145 modulated the differentiation of BMSCs into SMLCs in vivo . The apoptosis of the CCSMCs and the expression of cell α-actin were modulated by miR-145/KLF4 axis though paracrine. KLF4: Krüppel-like factor 4, miR-145: microRNA-145, BMSCs: bone marrow-derived mesenchymal stem cells, SRF: serum response factor, EdU: 5-ethynyl-2′-deoxyuridine, SMLCs: smooth muscle-like cells, CCSMCs: corpus cavernosum smooth muscle cells, ICI: intracavernous injection, CD31: platelet endothelial cell adhesion molecule-1.

Article Snippet: The lentivirus-miR-145-green fluorescent proteins (GFP) (Hanbio Biotechnology ) and primary BMSCs (Cyagen Biosciences) derived from rats were prepared.

Techniques: Modification, In Vitro, Migration, In Vivo, Expressing, Derivative Assay, Injection

Journal: ACS Biomaterials Science & Engineering

Article Title: Rat Calvarial Bone Regeneration by 3D-Printed β-Tricalcium Phosphate Incorporating MicroRNA-200c

doi: 10.1021/acsbiomaterials.0c01756

Figure Lengend Snippet: Collagen incorporating miR-200c increased miR-200c expression and osteogenic differentiation of rat and human BMSCs seeded on 3D-printed β-TCP scaffolds. (A) Relative expression levels of miR-200c from rBMSCs cultured as a monolayer for 3 days with different concentrations of pDNA encoding miR-200c or empty vector control. (B) Relative expression levels of miR-200c from rBMSCs 10 days after seeding onto β-TCP scaffolds with different treatments. (C, D) Relative expression levels of miR-200c from hBMSCs 3 days (C) and 7 days (D) after seeding onto β-TCP scaffolds with different treatments. (E, F) Normalized fold change of Runx2 transcript from hBMSCs 3 days (E) and 7 days (F) after seeding. (G, H) Normalized fold change of OCN (G) and OPG (H) transcripts in hBMSCs 7 days after seeding onto β-TCP scaffolds with different treatments. Column means that do not share a letter are statistically significantly different using the post hoc Tukey’s HSD test ( p < 0.05; performed in triplicate).

Article Snippet: Primary human BMSCs (hBMSCs; StemCells, Newark, CA) were cultured and expanded with completed minimum essential medium (MEM-α) supplemented with 10% FBS and 1% PS.

Techniques: Expressing, Cell Culture, Plasmid Preparation, Control

Journal: Journal of Nanobiotechnology

Article Title: Supramolecular delivery of senolytics enables targeted anti-senescence therapy and accelerated fracture healing

doi: 10.1186/s12951-026-04138-2

Figure Lengend Snippet: Asp 10 SAC4A enables hypoxia-activated senolysis and promotes osteogenic differentiation in vitro. A ) Heatmap of combination index (CI) values at different D: Q ratios and fraction affected (Fa) levels, showing synergistic effects of Dasatinib and Quercetin (DQ). B ) Cell viability curves of normal BMSCs, senescent BMSCs (Sn-BMSCs), and Sn-BMSCs under hypoxia treated with vehicle, free DQ, Asp 10 SAC4A, or DQ@Asp 10 SAC4A. C ) Representative SA-β-Gal staining images of senescent cells treated with different formulations under normoxic and hypoxic conditions, and quantification of SA-β-Gal-positive area (%) ( n = 4/group). Scale bar: 100 μm. D–F ) Representative Western blot images ( D ) and quantitative analyses ( E , F ) of senescence markers P16 and P21 expression in different treatment groups ( n = 3/group). G ) Representative alkaline phosphatase (ALP, upper panel) and Alizarin Red staining (ARS, lower panel) images demonstrating osteogenic differentiation after indicated treatments under normoxic and hypoxic conditions. H–J ) Representative Western blot images ( H ) and quantitative analyses ( I , J ) showing protein expression levels of osteogenic markers RUNX2 and osteopontin (OPN) ( n = 3/group). (Data are presented as mean ± SD; * P < 0.05, ** P < 0.01, *** P < 0.001; n = 3–4/group)

Article Snippet: Primary mouse bone marrow mesenchymal stem cells (BMSCs) were purchased from Servicebio (Catalog number STCC6011P).

Techniques: In Vitro, Staining, Western Blot, Expressing