Figure 3.

Oxidative stress, alkaline phosphatase (ALP) activity, and osteogenic gene assays for the first part of the experiment revealed that high glucose-induced ROS overproduction led to impaired osteogenic differentiation of titanium-surfaced BMSCs. (a) Oxidative stress analysis in the first part of the experiment revealed that a high-glucose environment induced excessive ROS generation in BMSCs cultured on titanium surfaces. (b) Quantitative analysis of fluorescence intensity for oxidative stress detection in the first part of the experiment. ⁣^∗p < 0.05; ⁣^∗∗p < 0.01; ⁣^∗∗∗p < 0.001. (c, d) ALP activity assays in the first part of the experiment revealed that excessive ROS production induced by high glucose resulted in reduced ALP activity in BMSCs cultured on titanium surfaces. ⁣^∗p < 0.05; ⁣^∗∗p < 0.01; ⁣^∗∗∗p < 0.001. (e) Quantitative real-time PCR (qRT-PCR) analysis of β-catenin and CaMK II genes revealed that excessive ROS production induced by high glucose activated the expression of the CaMK II signaling molecule while inhibiting the expression of the β-catenin signaling molecule. ⁣^∗p < 0.05; ⁣^∗∗p < 0.01; ⁣^∗∗∗p < 0.001. (f) qRT-PCR analysis of osteogenic genes demonstrated that excessive ROS production induced by high glucose resulted in decreased expression of osteogenic genes in BMSCs cultured on titanium surfaces. ⁣^∗p < 0.05; ⁣^∗∗p < 0.01; ⁣^∗∗∗p < 0.001.