Characterization and Study of Gene Expression Profiles of Human Periodontal Mesenchymal Stem Cells in Spheroid Cultures by Transcriptome Analysis

Figure 4

Enhancement of osteogenesis in hPDLMSC spheroid treated with NR4A2 siRNA through ALP activation. (a, b) Confirmation of silencing of NR4A2 by siRNA in spheroid-cultured hPDLMSC. NR4A2 siRNA decreased mRNA expression of NR4A2 to about 30%. Expression of NR4A2 also was reduced by siNR4A2. (c) Effects of siNR4A2 on OCT4, NANOG, and SOD2 mRNA expression in spheroid hPDLMSCs. The expression of OCT4, NANOG, and SOD2 mRNA was examined by real-time RT-PCR. (compared with control). (d) Nodule formation of hPDLMSC spheroid treated with siNR4A2 or control in OIM condition. Cells were transfected with siNR4A2 or control and then cultured for 12 days in osteoinductive medium and stained with alizarin red. (e) Effects of siNR4A2 on ALP, OPN, Col1, and RUNX2 mRNA expression in spheroid hPDLMSCs cultured with OIM. On day 7, the expression of ALP, OPN, Col1, and RUNX2 mRNA was examined by real-time RT-PCR. (compared with control). (f) Effects of siNR4A2 on ALP activity in spheroid hPDLMSCs cultured with OIM. On day 7, ALP activity was measured. (compared with control). (g) Effect of siNR4A2 on proliferation in spheroid hPDLMSCs on days 0, 3, and 7. All experiments were performed with samples from four donors.

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