Fig 1: The expression of calcification inhibitor was decreased in condylar cartilage, while the expression of calcification promotors was increased. A The number of MGP-positive chondrocytes and the mRNA expression of Mgp were decreased in the condylar cartilage of UAC rats (n = 6). B The number of TNAP-positive chondrocytes and the mRNA expression of Tnap were decreased in the condylar cartilage of UAC rats (n = 6). C The number of NPP1-positive chondrocytes and the mRNA expression of Npp1 were decreased in the condylar cartilage of UAC rats (n = 6). Scale bar = 100 μm. Control, control group; UAC, unilateral anterior crossbite group; 4w, 4 weeks; 8w, 8 weeks; 12w, 12 weeks. All data are presented as means ± SD. The data of compared groups were from the populations of Gaussian distribution and consistent with homogeneity of variance. Comparisons between the control and UAC groups of the same timepoint were performed by Student’s t test. **P < 0.01, compared with the age-matched control group
Fig 2: Inhibition of exosome formation alleviated calcification of condylar chondrocytes induced by FFSS. A Inhibition of exosome formation with GW4869 decreased the number of calcified nodules after FFSS stimulation in cultured condylar chondrocytes (n = 6). B GW4869 decreased exosome formation by condylar chondrocytes stimulated with FFSS (n = 6). C Inhibition of exosome formation did not affect the changes in protein levels in exosomes induced by FFSS (n = 6). Scale bar = 100 μm. All data are presented as means ± SD. Comparisons among groups were performed by one-way ANOVA, and it was followed by Tukey’s post hoc tests to evaluate the statistical significance in groups with the control group (0 dyn/cm2 group), respectively. *P<0.05, **P < 0.01, compared with the control group (0 dyn/cm2 group). D Supplementation with exogenous MGP or inhibition of TNAP and NPP1 effectively decreased the numbers of calcified nodules in condylar chondrocytes stimulated with FFSS (n = 6). Scale bar = 100 μm. Comparisons among groups were performed by one-way ANOVA, and it was followed by Tukey’s post hoc tests to evaluate the statistical significance in groups with the 16 dyn/cm2 group respectively. *P < 0.05, **P < 0.01, compared with the 16 dyn/cm2 group. E Exosomes from condylar chondrocytes stimulated with FFSS promoted the formation of calcified nodules in cultured condylar cartilage (n = 6). Scale bar = 100 μm. Comparisons among groups were performed by one-way ANOVA, and it was followed by Tukey’s post hoc tests to evaluate the statistical significance in groups with the control group (0 dyn/cm2 group), respectively. The data of compared groups were from the populations of Gaussian distribution and consistent with homogeneity of variance. *P < 0.05, **P < 0.01, compared with the control group (0 dyn/cm2 group)
Fig 3: FFSS promoted calcification and exosome formation in cultured condylar chondrocytes and changed the protein levels in exosomes. A FFSS promoted the formation of calcified nodules in cultured condylar chondrocytes (n = 6). Scale bar = 100 μm. B FFSS promoted the exosome formation of cultured condylar chondrocytes (n = 6). C FFSS changed the protein levels in exosomes, as demonstrated by decreased expression of MGP and increased expression of TNAP and NPP1 (n = 6). All data are presented as means ± SD. The data of compared groups were from the populations of Gaussian distribution and consistent with homogeneity of variance. Comparisons among groups were performed by one-way ANOVA, and it was followed by Tukey’s post hoc tests to evaluate the statistical significance in groups with the control group (0 dyn/cm2 group) respectively. *P < 0.05, **P < 0.01, compared with the control group (0 dyn/cm2 group)
Supplier Page from Abcam for Recombinant Human MGP protein (denatured) (His tag N-Terminus)