Next-generation Sequencing Identifies Articular Cartilage and Subchondral Bone Mirnas after ESWT on Early Osteoarthritis Knee C.-J. Wang, J.-H. Cheng,

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Next-generation Sequencing Identifies Articular Cartilage and Subchondral Bone Mirnas after ESWT on Early Osteoarthritis Knee C.-J. Wang, J.-H. Cheng, S.-H. Su, C.-Y. Huang, S.-L. Hsu Osteoarthritis and Cartilage Volume 25, Pages S321-S322 (April 2017) DOI: 10.1016/j.joca.2017.02.539 Copyright © 2017 Terms and Conditions

Figure 1: Molecular functions of predicted target genes of miRNAs differentially expressed in articular cartilage and subchondral bone. The diagrams of (A) articular cartilage and (B) subchondral bone showed the proportion of modulated miRNAs target genes belonging to the most significant functional annotations derived from the UniProt database. Osteoarthritis and Cartilage 2017 25, S321-S322DOI: (10.1016/j.joca.2017.02.539) Copyright © 2017 Terms and Conditions

Figure 2: The histologic analysis of cartilage and subchondral bone in a rat model of early OA. Safranin O staining of sagittal sections of the tibia medial compartment and proteoglycan of cartilage (red) and subchondral bone (blue) was shown. Black arrows indicated loss of proteoglycan at 4 weeks post-surgery. Scale bar: 50 μm. Osteoarthritis and Cartilage 2017 25, S321-S322DOI: (10.1016/j.joca.2017.02.539) Copyright © 2017 Terms and Conditions

Figure 3: Hierarchical clustering of the miRNA identified in each expression of library pattern from subchondral bone. The miRNAs were analysis from subchondral bone of sham (blue), OA (orange) and OA+SW (yellow). The color in clustering was greed=down regulation, and red= up regulation. Total miRNA was 69. Osteoarthritis and Cartilage 2017 25, S321-S322DOI: (10.1016/j.joca.2017.02.539) Copyright © 2017 Terms and Conditions

Figure 4: The profile of overall miRNAs expression changes Figure 4: The profile of overall miRNAs expression changes. (A) The differential expression levels of miRNA in library (Bone and cartilage) versus treatment groups (Sham, OA and OA+SW). Each shape in the 3D visualization represents a group of miRNAs. Principal component analysis (PCA) captured 80.1% of the variation observed in the experiment in the first three principal components (PC). (B) The source of the variation plot showed that the expression of miRNA in treatment groups of factor accounted for major of the variation. Osteoarthritis and Cartilage 2017 25, S321-S322DOI: (10.1016/j.joca.2017.02.539) Copyright © 2017 Terms and Conditions

Figure 5: MiRNA expression patterns were evaluated by Quantitative RT-PCR. The rno-miR-199a-3p, rno-miR- 181a-5p, rno-miR-140-3p and rno-miR-27b-3p were measured from articular cartilage (A) and subchondral bone (B). The expression profiles of each miRNA matched the tested probes at least three times repeat. The values were the mean and SEM of the miRNA expression levels in 4 animals, as determined by ΔCt analysis, normalized to GAPDH expression, and relative to the expression levels of sham control. ** p<0.01 (versus sham control). Osteoarthritis and Cartilage 2017 25, S321-S322DOI: (10.1016/j.joca.2017.02.539) Copyright © 2017 Terms and Conditions