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•We explore the molecular mechanism by coupling iTRAQ with nano-LC–MS/MS.•Ca2+ could induce osteogenic differentiation of BM-hMSCs in serum-free medium.•MAPKs pathway might participate in the osteogenic differentiation of BM-hMSCs.•Some novel proteins were discovered in osteogenic differentiation of BM-hMSCs.
Human mesenchymal stem cells-bone marrow (BM-hMSCs) are considered as the most suitable seed cells for bone tissue engineering. Calcium ions (Ca2+) forms an important component of a number of commercial bone substitutes and support materials. For efficient bone tissue engineering, it is crucial to explore the effect of extracellular Ca2+ on the growth and differentiation of BM-hMSCs, and to understand their molecular mechanisms. Therefore, in the present study, BM-hMSCs were cultivated in serum free growth medium or serum free growth medium with additional 4 or 6mM Ca2+ for 3weeks, following which, the proliferation and osteoblastic differentiation of these cells were evaluated. Differentially expressed proteins were established using iTRAQ labeling coupled with nano-LC–MS/MS. Our data revealed that Ca2+ significantly promoted the proliferation of BM-hMSCs in the early stage. Furthermore, Ca2+ showed osteoinduction properties. MAPKs signaling pathway might participate in the osteogenic differentiation of BM-hMSCs caused by Ca2+. Certain newly found proteins could be potentially important for the osteogenic differentiation of BM-hMSCs and may be associated with osteogenesis.