Fig. 3
Validation of selected circulating microRNAs (miRNAs) using individual quantitative real-time reverse transcriptase PCR (qPCR) assays. (A) The results of gene stability analysis of qPCR assay targets using the geNorm algorithm. Because no miRNA had high expression stability (M < 0.5), the two most stable miRNAs (miR-23a-5p and miR-221-3p) with an intermediate level of stability (0.5 < M < 1) were selected as reference genes. (B) Box plots representing the expression levels of each miRNA in patients with medication-related osteonecrosis of the jaw (MRONJ) compared to healthy controls. Among the target genes selected for the second phase, the expression levels of five genes were significantly altered in the MRONJ group compared to the control (CON) group (CNRQ, calibrated normalized relative quantity, *Q < 0.05, **Q < 0.001, ***Q < 0.0001). (C) Heatmap illustrating the differential expression of the five miRNAs between the MRONJ and control groups. Average linkage clustering based on Euclidean distances of expression values effectively distinguished the MRONJ group from the control group.
