Oxidative stress-induced circSOD2 inhibits osteogenesis through sponging miR-29b in metabolic-associated fatty liver disease

Figure 4 circSOD2 inhibits osteogenesis. A-D: MRNA expression of osteogenic markers, including alkaline phosphatase (ALP), runt-related transcription factor 2, osteocalcin, and collagen type 1 alpha 1 in bone marrow mesenchymal stem cells (BMSCs) incubated with osteogenic medium for 7 days after transfection of AD-circSOD2 or si-circSOD2; E: ALP staining at 7 days and Alizarin Red staining at 14 days, demonstrating ALP activity and calcium deposition, respectively; F and G: Quantification of ALP activity and calcium deposition; H: Representative micro-computed tomography imaging of bone formation in rat calvarial critical-size defect mode at 8 weeks. Rats (n = 25) were randomly assigned to five groups: Blank (Con), overexpression (AD-circSOD2), overexpression control (AD-NC), knockdown (Sh-circSOD2), knockdown control (Sh-NC); I: Trabecular bone volume per tissue volume from histomorphometric analysis in the calvarial model (n = 5); J: Quantitative analysis of bone mineral density in the calvarial rat model (n = 5). ^aP < 0.05; ^bP < 0.01; ALP: Alkaline phosphatase; RUNX2: Runt-related transcription factor 2; OCN: Osteocalcin; COL1A1: Collagen type 1 alpha 1; Con: Control; BV/TV: Bone volume per tissue volume.