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The effect of TNFRSF11B-OE-CM on the differentiation of osteoclasts. THP-1 cells were incubated with PMA for 3 days, followed by culture in the presence of M-CSF (100 ng/mL), RANKL (100 ng/mL), and either control-CM or TNFRSF11B-OE-CM for an additional 12 days. (A, B) Representative images of TRAP staining and corresponding statistical analysis illustrate the number of TRAP-positive multinucleated cells with differentiated osteoclasts marked by red arrows. (C) TRAP activity was quantified in THP-1 cells treated with control-CM or TNFRSF11B-OE-CM. (D) qRT-PCR analysis was conducted to assess the expression levels of osteoclast differentiation-related marker genes in each group. Blank represents the uninduced group. Comparisons were made relative to the M-CSF and RANKL (M + R) stimulation group, # p < 0.05, ## p < 0.01; compared with the control-CM group, * p < 0.05; ** p < 0.01

Journal: Journal of Orthopaedic Surgery and Research

Article Title: TNFRSF11B-modified umbilical cord mesenchymal stem cells as a novel strategy for bone-related diseases by suppressing osteoclast activity

doi: 10.1186/s13018-025-05850-9

Figure Lengend Snippet: The effect of TNFRSF11B-OE-CM on the differentiation of osteoclasts. THP-1 cells were incubated with PMA for 3 days, followed by culture in the presence of M-CSF (100 ng/mL), RANKL (100 ng/mL), and either control-CM or TNFRSF11B-OE-CM for an additional 12 days. (A, B) Representative images of TRAP staining and corresponding statistical analysis illustrate the number of TRAP-positive multinucleated cells with differentiated osteoclasts marked by red arrows. (C) TRAP activity was quantified in THP-1 cells treated with control-CM or TNFRSF11B-OE-CM. (D) qRT-PCR analysis was conducted to assess the expression levels of osteoclast differentiation-related marker genes in each group. Blank represents the uninduced group. Comparisons were made relative to the M-CSF and RANKL (M + R) stimulation group, # p < 0.05, ## p < 0.01; compared with the control-CM group, * p < 0.05; ** p < 0.01

Article Snippet: The measurement of TRAP activity utilized TRAP activity assay kits (Elabscience, Cat. No. E-BC-K871-M, Wuhan, China), with absorbance readings obtained at 405 nm via microplate reader.

Techniques: Incubation, Control, Staining, Activity Assay, Quantitative RT-PCR, Expressing, Marker