Journal: Materials Today Bio

Article Title: Hepatocyte growth factor in biosheets promotes autonomous regeneration of cutaneous tissue after transplantation onto a full-thickness skin defect

doi: 10.1016/j.mtbio.2026.102969

Figure Lengend Snippet: Analysis of surface markers and multipotency of stromal cells in biosheets (A) Representative flow cytometry profiles of cells isolated from 2w-biosheets and identification of a population of quadruple-positive cells (P7). (B) The ratio of quadruple-positive cells (P7) to stromal cells (P5) in the biosheets and fascia. (C-E) Representative images of alkaline phosphatase (ALP) staining and Alizarin red staining (C), oil red staining (D), Toluidine blue (TB) staining and Safranine O (SO) staining (E) of stromal cells isolated from biosheets and subjected to osteogenic, adipogenic and chondrogenic differentiation. (F-H) Expression levels of osteogenic (F), adipogenic (G), and chondrogenic (H) marker genes. (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 cells formed pellets after 3 days of culture, and the medium was replaced with MSC Chondrogenic Differentiation Medium (C-28012, PromoCell).

Techniques: Flow Cytometry, Isolation, Staining, Expressing, Marker

Journal: Molecular medicine reports

Article Title: MicroRNA‑125b suppresses the proliferation and osteogenic differentiation of human bone marrow‑derived mesenchymal stem cells.

doi: 10.3892/mmr.2014.2024

Figure Lengend Snippet: Figure 1. Comparison of the proliferation and osteogenic differen tiation of hBMSCs isolated from osteoporotic patients and normal subjects. (A) Growth and viability of hBMSCs were determined by the MTT assay after cells were cultured for 2, 4, 6, 8 and 10 days; (B) hBMSCs at passage 3 were cultured in osteogenic medium for 14 days, followed by staining and assessment of ALP activity. Data were analyzed using the Student's t‑tests; (C) quantitative polymerase chain reaction analysis of miR‑125b expression in hBMSCs. miR‑125b expression levels were increased in hBMSCs isolated from elderly patients. Data were subjected to Student's t‑tests. Error bars represent the mean ± standard deviation of three independent experiments. *P<0.05. hBMSCs, human bone marrow‑derived mesenchymal stem cells; ALP, alkaline phosphatase; miR, micro RNA; OD, optical density.

Article Snippet: On every third day, the medium was replaced with osteogenic differentiation medium (10% fetal bovine serum, Hyclone; 100 nM dexamethasone, 45 mM L-ascorbic acid and 10 mM β-glycerophosphate; Sigma, St. Louis, MO, USA).

Techniques: Comparison, Isolation, MTT Assay, Cell Culture, Staining, Activity Assay, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

Journal: Molecular medicine reports

Article Title: MicroRNA‑125b suppresses the proliferation and osteogenic differentiation of human bone marrow‑derived mesenchymal stem cells.

doi: 10.3892/mmr.2014.2024

Figure Lengend Snippet: Figure 3. (A) Overexpression of miR‑125b suppressed the osteogenic differentiation of hBMSCs. Osteoblast differentiation of hBMSCs was induced by osteogenic differentiation medium and hBMSCs were collected 14 days after osteogenic induction. qPCR analysis measured the expression of Runx‑2, ALP, OC and COL1α1 in hBMSCs. qPCR data were subjected to Student's t‑tests. Error bars represent the mean ± standard deviation of three independent experi ments. *P<0.05. (B) ALP staining was performed after 14 days of culture to detect ALP activity and Alizarin Red staining was performed after 21 days to evaluate mineralized bone matrix formation. (C) ALP activity of hBMSCs was determined 14 days after osteogenic induction by p‑Nitrophenyl Phosphate Liquid Substrate System and absorbance was measured at 405 nm. Data were subjected to Student's t‑test. *P<0.05. hBMSCs, human bone marrow‑derived mesenchymal stem cells; qPCR, quantitative polymerase chain reaction; Runx‑2, Runt‑related transcription factor‑2; ALP, alkaline phosphatase; OC, osteo calcin; COL1α1, collagen type I α1; NC, negative control; miR, micro RNA; OD, optical density.

Article Snippet: On every third day, the medium was replaced with osteogenic differentiation medium (10% fetal bovine serum, Hyclone; 100 nM dexamethasone, 45 mM L-ascorbic acid and 10 mM β-glycerophosphate; Sigma, St. Louis, MO, USA).

Techniques: Over Expression, Expressing, Standard Deviation, Staining, Activity Assay, Real-time Polymerase Chain Reaction, Negative Control

Journal: Cells

Article Title: Melatonin Attenuates Spinal Cord Injury in Mice by Activating the Nrf2/ARE Signaling Pathway to Inhibit the NLRP3 Inflammasome

doi: 10.3390/cells11182809

Figure Lengend Snippet: The oxidative damage scavenging effect of melatonin in neuron cells. ( A ) The survival rates of PC12 cells that received different concentrations of melatonin treatment were detected using a CCK-8 kit (n = 8). ( B ) The ROS produced in PC12 cells that received different treatments were detected by ROS staining (n = 6, scale bar = 100 µm). ( C , D ) The mitochondrial membrane potential in PC12 cells treated with drugs was detected by JC-1 staining (n = 8, scale bar = 100 µm). ( E – G ) The contents of SOD and MDA and the activity of GSH-PX were detected using the corresponding kits in PC12 cells treated with different treatment factors (n = 10). Data are shown as the means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of IL-1β and IL-18 were measured in the PC12 cell culture medium using ELISA kits according to the manufacturer’s instructions (Elabscience, China).

Techniques: CCK-8 Assay, Produced, Staining, Membrane, Activity Assay

Journal: Cells

Article Title: Melatonin Attenuates Spinal Cord Injury in Mice by Activating the Nrf2/ARE Signaling Pathway to Inhibit the NLRP3 Inflammasome

doi: 10.3390/cells11182809

Figure Lengend Snippet: Melatonin suppresses the NLRP3 inflammasome in PC12 cells treated with H2O2 by activating the Nrf2/ARE pathway. ( A – I ) Western blot detection and statistical analysis of NLRP3-inflammasome-related proteins NLRP3, caspase-1, ASC, and IL-1β; and antioxidant-stress-related proteins nuclear Nrf2, HO-1, and NQO-1 in PC12 cells from each group (n = 6). ( J , K ) The levels of IL-1β and IL-18 in the supernatant of PC12 cells were determined by ELISA kits (n = 6). Data are shown as the means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of IL-1β and IL-18 were measured in the PC12 cell culture medium using ELISA kits according to the manufacturer’s instructions (Elabscience, China).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Melatonin Attenuates Spinal Cord Injury in Mice by Activating the Nrf2/ARE Signaling Pathway to Inhibit the NLRP3 Inflammasome

doi: 10.3390/cells11182809

Figure Lengend Snippet: Immunofluorescence double staining of Nrf2 and NLRP3 in PC12 cells treated with H2O2. Immunofluorescence double staining was performed to test the protein level of Nrf2 ( A , B ) and NLRP3 ( C , D ) in PC12 cells from each group (n = 8, scale bar = 100 µm). Data are shown as the means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of IL-1β and IL-18 were measured in the PC12 cell culture medium using ELISA kits according to the manufacturer’s instructions (Elabscience, China).

Techniques: Immunofluorescence, Double Staining

Journal: International Journal of Nanomedicine

Article Title: Schisandrae fructus enhances myogenic differentiation and inhibits atrophy through protein synthesis in human myotubes

doi: 10.2147/IJN.S101299

Figure Lengend Snippet: SF-induced differentiation in HSkMCs. Notes: ( A ) SF was treated with various concentrations (0.1 μg/mL, 0.5 μg/mL, 1 μg/mL, 5 μg/mL, and 10 μg/mL) in growth medium. Two days after treatment, HSkMCs were counted. As a result, SF did not induce proliferation. ( B and C ) SF (0.1 μg/mL, 0.5 μg/mL, 1 μg/mL, 5 μg/mL, and 10 μg/mL) was treated with DM and changed with fresh DM every 2 days. HSkMCs were photographed three times every 2 days. For observing differentiation efficiency, fusion index and myotube area were analyzed. ( D ) Eight days after differentiation induction with SF, HSkMCs were photographed. A total of 0.5 μg/mL of SF significantly induced more differentiation. All data represented mean ± SEM (n=3). *Symbol indicates P <0.05 compared to control. Abbreviations: DM, differentiation medium; HSkMCs, human skeletal muscle cells; SF, Schisandrae fructus.

Article Snippet: Differentiation of HSkMCs was induced by HSkMC differentiation medium (DM) (C-23061; PromoCell, Heidelberg, Germany).

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Schisandrae fructus enhances myogenic differentiation and inhibits atrophy through protein synthesis in human myotubes

doi: 10.2147/IJN.S101299

Figure Lengend Snippet: SF increased protein synthesis through mTOR/P70S6K and 4E-BP1 signaling. Notes: ( A ) SF (0.5 μg/mL) was treated with differentiation medium. HSkMCs’ differentiation was induced for 1 day, 2 days, and 4 days. Phosphorylation and expression of mTOR, P70S6K, and 4E-BP1 were observed for Western blot. GAPDH expression was analyzed to identify equal loading. ( B – D ) Levels of mTOR activation (phosphorylation) were normalized to the levels of GAPDH. Phosphorylation of P70S6K and 4E-BP1 was normalized to the levels of each total protein. All data represented mean ± SEM (n=3). *Symbol indicates P <0.05. Abbreviations: 4E-BP1, 4E-binding protein1; HSkMCs, human skeletal muscle cells; mTOR, mammalian target of rapamycin; p-4E-BP1, phosphorylated 4E-BP1; p-P70S6K, phosphorylated P70S6K; P70S6K, 70 kDa ribosomal protein S6 kinase; SF, Schisandrae fructus; SEM, standard error of mean; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Differentiation of HSkMCs was induced by HSkMC differentiation medium (DM) (C-23061; PromoCell, Heidelberg, Germany).

Techniques: Expressing, Western Blot, Activation Assay, Binding Assay

Journal: International Journal of Nanomedicine

Article Title: Schisandrae fructus enhances myogenic differentiation and inhibits atrophy through protein synthesis in human myotubes

doi: 10.2147/IJN.S101299

Figure Lengend Snippet: SF inhibited muscle atrophy through increasing protein synthesis. Notes: At 2 days of differentiation, atrophy of myotubes was induced by 100 μM DEX for 4 days, and DM was changed to fresh DM with SF every 2 days. ( A – C ) In the last 6 days after induction of differentiation, HSkMCs were photographed three times per group. For observing differentiation efficiency, fusion index and myotube area were analyzed. All data represented mean ± SEM (n=3). *Symbol indicates P <0.05 compared to control. # Symbol represents P <0.05 compared to DEX treatment alone. ( D ) Myotubes were fluorescence stained with anti-MYH (red) and DAPI (blue), which was observed as a marker of late differentiation. ( E and F ) Examples of representative Western blot were shown for MYH, p-P70S6K, P70S6K p-4E-BP1, 4E-BP1, GAPDH, and MuRF1. Levels of MYH, MuRF1, p-P70S6K, and p-4E-BP1 were normalized to the levels of GAPDH or total protein. Abbreviations: DEX, dexamethasone; DM, differentiation medium; 4E-BP1, 4E-binding protein1; HSkMCs, human skeletal muscle cells; MuRF1, muscle RING finger 1; MYH, myosin heavy chain; p-4E-BP1, phosphorylated eukaryotic translation initiation factor 4E-binding protein 1; p-P70S6K, phosphorylated P70S6K; P70S6K, 70 kDa ribosomal protein S6 kinase; SF, Schisandrae fructus; SEM, standard error of mean; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Differentiation of HSkMCs was induced by HSkMC differentiation medium (DM) (C-23061; PromoCell, Heidelberg, Germany).

Techniques: Fluorescence, Staining, Marker, Western Blot, Binding Assay