Cell Culture:Article Title: Pure platelet-rich plasma delays intervertebral disc degeneration by activating SIRT1-mediated autophagy in nucleus pulposus cells
Article Snippet: .. All groups were cultured for a total of 48 h. The experimental conditions were as follows: (1) the control group, which was treated with complete medium (DMEM/F12: fetal bovine serum = 9:1) for 48 h without any stimulation; (2) the IVDD group, which was treated with IL-1β (10 ng/mL, MedChemExpress, HY- P73150 , USA) for 48 h; (3) the SRT1720 group, which was treated with IL-1β (10 ng/mL) for 24 h, followed by co-incubation with IL-1β (10 ng/mL) and SRT1720 (1 μmol/L, MedChemExpress, HY-10532, USA), a selective SIRT1 activator, for an additional 24 h; (4) the EX527 group, which was treated as in the SRT1720 group but with the addition of EX-527 (5 μmol/L, MedChemExpress, HY-15452, USA), a selective SIRT1 inhibitor, during the second 24 h period; (5) the PRP group, which was treated with IL-1β (10 ng/mL) for 24 h, then co-incubated with IL-1β (10 ng/mL) and 200 μL of P-PRP for another 24 h, achieving a final PRP concentration of 10% (v/v); and (6) the PRP + EX527 group, which was treated identically to the PRP group, with the addition of EX-527 (5 μmol/L) during the final 24 h. ..
Article Title: Pure platelet-rich plasma delays intervertebral disc degeneration by activating SIRT1-mediated autophagy in nucleus pulposus cells.
Article Snippet: .. All groups were cultured for a total of 48 h. The experimental conditions were as follows: (1) the control group, which was treated with complete medium (DMEM/F12: fetal bovine serum = 9:1) for 48 h without any stimulation; (2) the IVDD group, which was treated with IL-1β (10 ng/mL, MedChemExpress, HY-P73150, USA) for 48 h; (3) the SRT1720 group, which was treated with IL-1β (10 ng/ mL) for 24 h, followed by co-incubation with IL-1β (10 ng/mL) and SRT1720 (1 μmol/L, MedChemExpress, HY-10532, USA), a selective SIRT1 activator, for an additional 24 h; (4) the EX527 group, which was treated as in the SRT1720 group but with the addition of EX-527 (5 μmol/L, MedChemExpress, HY-15452, USA), a selective SIRT1 inhibitor, during the second 24 h period; (5) the PRP group, which was treated with IL-1β (10 ng/mL) for 24 h, then co-incubated with IL-1β (10 ng/mL) and 200 μL of P-PRP for another 24 h, achieving a final PRP concentration of 10% (v/v); and (6) the PRP + EX527 group, which was treated identically to the PRP group, with the addition of EX-527 (5 μmol/L) during the final 24 h. Mitochondrial membrane potential (ΔΨm) assay NPCs were seeded in 6-well plates and treated as described previously. .. The cells were washed three times with PBS and dark incubated with 2 μM JC-1 dye (Beyotime, C2006, CN) at 37 °C for 20 min.
Control:Article Title: Pure platelet-rich plasma delays intervertebral disc degeneration by activating SIRT1-mediated autophagy in nucleus pulposus cells
Article Snippet: .. All groups were cultured for a total of 48 h. The experimental conditions were as follows: (1) the control group, which was treated with complete medium (DMEM/F12: fetal bovine serum = 9:1) for 48 h without any stimulation; (2) the IVDD group, which was treated with IL-1β (10 ng/mL, MedChemExpress, HY- P73150 , USA) for 48 h; (3) the SRT1720 group, which was treated with IL-1β (10 ng/mL) for 24 h, followed by co-incubation with IL-1β (10 ng/mL) and SRT1720 (1 μmol/L, MedChemExpress, HY-10532, USA), a selective SIRT1 activator, for an additional 24 h; (4) the EX527 group, which was treated as in the SRT1720 group but with the addition of EX-527 (5 μmol/L, MedChemExpress, HY-15452, USA), a selective SIRT1 inhibitor, during the second 24 h period; (5) the PRP group, which was treated with IL-1β (10 ng/mL) for 24 h, then co-incubated with IL-1β (10 ng/mL) and 200 μL of P-PRP for another 24 h, achieving a final PRP concentration of 10% (v/v); and (6) the PRP + EX527 group, which was treated identically to the PRP group, with the addition of EX-527 (5 μmol/L) during the final 24 h. ..
Article Title: Pure platelet-rich plasma delays intervertebral disc degeneration by activating SIRT1-mediated autophagy in nucleus pulposus cells.
Article Snippet: .. All groups were cultured for a total of 48 h. The experimental conditions were as follows: (1) the control group, which was treated with complete medium (DMEM/F12: fetal bovine serum = 9:1) for 48 h without any stimulation; (2) the IVDD group, which was treated with IL-1β (10 ng/mL, MedChemExpress, HY-P73150, USA) for 48 h; (3) the SRT1720 group, which was treated with IL-1β (10 ng/ mL) for 24 h, followed by co-incubation with IL-1β (10 ng/mL) and SRT1720 (1 μmol/L, MedChemExpress, HY-10532, USA), a selective SIRT1 activator, for an additional 24 h; (4) the EX527 group, which was treated as in the SRT1720 group but with the addition of EX-527 (5 μmol/L, MedChemExpress, HY-15452, USA), a selective SIRT1 inhibitor, during the second 24 h period; (5) the PRP group, which was treated with IL-1β (10 ng/mL) for 24 h, then co-incubated with IL-1β (10 ng/mL) and 200 μL of P-PRP for another 24 h, achieving a final PRP concentration of 10% (v/v); and (6) the PRP + EX527 group, which was treated identically to the PRP group, with the addition of EX-527 (5 μmol/L) during the final 24 h. Mitochondrial membrane potential (ΔΨm) assay NPCs were seeded in 6-well plates and treated as described previously. .. The cells were washed three times with PBS and dark incubated with 2 μM JC-1 dye (Beyotime, C2006, CN) at 37 °C for 20 min.
Concentration Assay:Article Title: Pure platelet-rich plasma delays intervertebral disc degeneration by activating SIRT1-mediated autophagy in nucleus pulposus cells
Article Snippet: .. All groups were cultured for a total of 48 h. The experimental conditions were as follows: (1) the control group, which was treated with complete medium (DMEM/F12: fetal bovine serum = 9:1) for 48 h without any stimulation; (2) the IVDD group, which was treated with IL-1β (10 ng/mL, MedChemExpress, HY- P73150 , USA) for 48 h; (3) the SRT1720 group, which was treated with IL-1β (10 ng/mL) for 24 h, followed by co-incubation with IL-1β (10 ng/mL) and SRT1720 (1 μmol/L, MedChemExpress, HY-10532, USA), a selective SIRT1 activator, for an additional 24 h; (4) the EX527 group, which was treated as in the SRT1720 group but with the addition of EX-527 (5 μmol/L, MedChemExpress, HY-15452, USA), a selective SIRT1 inhibitor, during the second 24 h period; (5) the PRP group, which was treated with IL-1β (10 ng/mL) for 24 h, then co-incubated with IL-1β (10 ng/mL) and 200 μL of P-PRP for another 24 h, achieving a final PRP concentration of 10% (v/v); and (6) the PRP + EX527 group, which was treated identically to the PRP group, with the addition of EX-527 (5 μmol/L) during the final 24 h. ..
Article Title: Pure platelet-rich plasma delays intervertebral disc degeneration by activating SIRT1-mediated autophagy in nucleus pulposus cells.
Article Snippet: .. All groups were cultured for a total of 48 h. The experimental conditions were as follows: (1) the control group, which was treated with complete medium (DMEM/F12: fetal bovine serum = 9:1) for 48 h without any stimulation; (2) the IVDD group, which was treated with IL-1β (10 ng/mL, MedChemExpress, HY-P73150, USA) for 48 h; (3) the SRT1720 group, which was treated with IL-1β (10 ng/ mL) for 24 h, followed by co-incubation with IL-1β (10 ng/mL) and SRT1720 (1 μmol/L, MedChemExpress, HY-10532, USA), a selective SIRT1 activator, for an additional 24 h; (4) the EX527 group, which was treated as in the SRT1720 group but with the addition of EX-527 (5 μmol/L, MedChemExpress, HY-15452, USA), a selective SIRT1 inhibitor, during the second 24 h period; (5) the PRP group, which was treated with IL-1β (10 ng/mL) for 24 h, then co-incubated with IL-1β (10 ng/mL) and 200 μL of P-PRP for another 24 h, achieving a final PRP concentration of 10% (v/v); and (6) the PRP + EX527 group, which was treated identically to the PRP group, with the addition of EX-527 (5 μmol/L) during the final 24 h. Mitochondrial membrane potential (ΔΨm) assay NPCs were seeded in 6-well plates and treated as described previously. .. The cells were washed three times with PBS and dark incubated with 2 μM JC-1 dye (Beyotime, C2006, CN) at 37 °C for 20 min.
Membrane:Article Title: Pure platelet-rich plasma delays intervertebral disc degeneration by activating SIRT1-mediated autophagy in nucleus pulposus cells.
Article Snippet: .. All groups were cultured for a total of 48 h. The experimental conditions were as follows: (1) the control group, which was treated with complete medium (DMEM/F12: fetal bovine serum = 9:1) for 48 h without any stimulation; (2) the IVDD group, which was treated with IL-1β (10 ng/mL, MedChemExpress, HY-P73150, USA) for 48 h; (3) the SRT1720 group, which was treated with IL-1β (10 ng/ mL) for 24 h, followed by co-incubation with IL-1β (10 ng/mL) and SRT1720 (1 μmol/L, MedChemExpress, HY-10532, USA), a selective SIRT1 activator, for an additional 24 h; (4) the EX527 group, which was treated as in the SRT1720 group but with the addition of EX-527 (5 μmol/L, MedChemExpress, HY-15452, USA), a selective SIRT1 inhibitor, during the second 24 h period; (5) the PRP group, which was treated with IL-1β (10 ng/mL) for 24 h, then co-incubated with IL-1β (10 ng/mL) and 200 μL of P-PRP for another 24 h, achieving a final PRP concentration of 10% (v/v); and (6) the PRP + EX527 group, which was treated identically to the PRP group, with the addition of EX-527 (5 μmol/L) during the final 24 h. Mitochondrial membrane potential (ΔΨm) assay NPCs were seeded in 6-well plates and treated as described previously. .. The cells were washed three times with PBS and dark incubated with 2 μM JC-1 dye (Beyotime, C2006, CN) at 37 °C for 20 min.
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