GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE343nnn/GSE343220/primaryOK200TranscriptomicsMus musculusExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE343220GEOGSEfalseSelf-assembled jawbone periosteal cell spheroids drive rapid osteogenic commitment through F-actin/NF-κB-mediated mechanotransductionThree-dimensional (3D) organoid has emerged as a revolutionary approach for studying ossification and bone regeneration. However, it remains unclear whether a 3D organoid system can trigger rapid osteogenic commitment in a physiologically relevant, induction-free condition. In this study, we established a scaffold-free, induction-free self-assembling spheroid model using mouse jawbone periosteum-derived cells (jb-PDCs). Remarkably, spheroid formation triggered a significant upregulation of osteogenic commitment markers Col1 at 12 hours, Sp7 at 24 hours, and Spp1 at 48 hours progenitor cells without exogenous inductive factors. Transcriptomic profiling revealed prominent enrichment of the NF-κB signaling pathway, and increased phosphorylation of the p65 subunit confirmed pathway activation. Pharmacological inhibition of NF-κB pathway abolished the upregulation of Col1, Sp7 and Spp1. In contrast, direct activation in conventional monolayer cultures recapitulated the same gene expression program. Furthermore, spheroid formation induced significant disassembly of filamentous actin, and pharmacological stabilization of actin filaments downregulated both p65 phosphorylation and osteogenic gene expression. These findings establish a mechanotransduction axis whereby three-dimensional self-assembly triggers actin cytoskeletal remodeling, leading to NF-κB activation and subsequent osteogenic commitment. Our work reveals that the physical event of cellular condensation intrinsically couples to an osteogenic priming program, providing new insights into how biophysical cues govern the early phases of bone formation, The jb-PDCs spheroid system established here also offers a unique platform for dissecting the mechanobiology of osteogenic initiation and may inform biomimetic strategies for craniofacial bone regeneration.2026/08/12GSE343220GSM9947963GSM9947961GSM9947962GSM9947960GSM9947958GSM99479599185343220Mus musculus