Fig. 2

Inflammatory cytokines cause mitochondrial dysfunction in PDLSCs during cementoblastic differentiation. The cells were incubated in normal α-MEM (Nor), medium with the inflammatory cytokines TNF-α and IL-1β (Infla), medium with the cementoblastic inducer EMD (EMD), or medium with both inflammatory cytokines and EMD (Infla-EMD). a The MMP was determined using a JC-1 probe (immunofluorescence staining; scale bar: 200 μm). b Quantification of the MMP (reflected by the relative ratio of red/green fluorescence intensity of JC-1). c Intracellular ATP contents determined by ATP assays. d mtDNA content determined by qRT-PCR. e Morphometric analysis of mitochondria by transmission electron microscopy (yellow arrows indicate healthy mitochondria, red arrows indicate swollen mitochondria; scale bar: 500 nm). f The OCR of PDLSCs determined using a Seahorse Bioscience XF Analyzer; arrows indicate the sequential injection of the ATPase inhibitor Oligo (1 μmol·L⁻¹), the uncoupling reagent carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, 1 μmol·L⁻¹) and inhibitors of the electron transport chain rotenone/antimycin (R/A, 2 μmol·L⁻¹). g Quantification of maximal respiration (differences between the maximum rate measurement after FCCP injection and the minimum rate measurement after R/A injection). h Quantification of cellular respiration (basic OCR value prior to Oligo injection). i Quantification of ATP production (difference between final rate measurement prior to Oligo injection and minimum rate measurement after Oligo injection). The data are shown as the mean ± SD for n from 3 to 8; *P < 0.05, **P < 0.01 and ***P < 0.001 indicate significant differences between the indicated columns