Bone regeneration depends on the coordinated interplay between mesenchymal stem cells (MSCs), immune-derived signals, and processes of extracellular matrix (ECM) remodeling. While most Toll-like receptors (TLRs) evoke pro-inflammatory responses that impair osteogenesis, the role of the atypical, immunomodulatory receptor TLR10 in MSC biology remains largely unexplored. Here, we investigated whether TLR10 serves as a regulatory node integrating innate immune signaling with vitamin D–driven osteogenic differentiation in human adipose-derived MSCs. ASC/TERT1 MSCs were engineered to overexpress or silence TLR10 using lentiviral vectors.