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Updated: Mar 19, 2026
Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice
Published on: January 6, 2018
Proenkephalin Inhibits the Osteogenic Differentiation of Bone Marrow-derived Mesenchymal Stem Cells
Objective:
To investigate the regulatory role of proenkephalin (PENK) on the osteogenic differentiation process of human bone marrow mesenchymal stem cells (hBMMSCs).
Methods:
PENK knockdown and overexpression in hBMMSCs were performed by using lentivirus containing the specific short hairpin RNA or whole coding region for PENK in vitro. The alkaline phosphatase level and mineralised nodule formation were observed by alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining and quantification. Meanwhile, the expression levels of osteogenic-related mRNA and protein were detected by qRT-PCR and Western blot.
Results:
Transfection of lentivirus in hBMMSCs was effectively confirmed by the green fluorescent protein-positive cells under fluorescence microscopy, and qRT-PCR and Western blot analyses verified the successful establishment of PENK knockdown and overexpression hBMMSCs. The results showed that knockdown of PENK effectively promoted the osteogenic differentiation of hBMMSCs, with enhanced ALP and ARS staining, and increased expression levels of osteogenic-related genes and proteins. Conversely, the overexpression of PENK inhibited the osteogenic differentiation of hBMMSCs.
Conclusion:
This finding identified PENK as a potential therapeutic target for bone regeneration of oral and maxillofacial bone defects.
