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Published on: May 4, 2018
CTP-CM enhances osteogenic differentiation of hPDLSCs via NF-κB pathway
Na Li1, Zehan Li1,2, Yanqiu Wang3
1Institute of Stomatology, Nanjing Medical University, Nanjing, China.
Objective:
The conditioned medium of calcined tooth powder (CTP-CM) is a type of biomimetic mineralized material and well contributing to bone healing and bone formation in vivo. However, little is known about the effect of CTP-CM on human periodontal ligament stem cells (hPDLSCs) as well as the underlying mechanisms.
Methods:
ALP activity assay was conducted to select the concentration with the highest ALP level, which was used for the following experiments. Cell proliferation was measured by cell counting kit-8 assay and flow cytometry analysis. Expression levels of osteogenic markers in CTP-CM-induced hPDLSCs were evaluated with real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunofluorescence staining, and Western blot. Mineralization of CTP-CM-induced hPDLSCs was evaluated by alizarin red staining. Furthermore, the involvement of NF-κB pathway was examined by immunofluorescence staining and Western blot.
Results:
20 μg/ml was selected for the further experiments. Functional studies demonstrated that CTP-CM exerted almost no influence on the proliferation of hPDLSCs and CTP-CM increased the osteogenic differentiation of hPDLSCs. Mechanistically, CTP-CM leads to activation of NF-κB signaling pathway. When treated with BMS345541, the osteogenic differentiation of CTP-CM-treated hPDLSCs was significantly attenuated.
Conclusion:
CTP-CM can promote the osteogenic differentiation of hPDLSCs via activating NF-κB pathway.
