Fig 2: Proposed mechanism of thermo-mechanotransduction process under low-oxygen tension condition(A) Integrated fluorescent intensities for SOX9 with representative confocal images of human articular chondrocytes seeded in GelMA-based hydrogels channels after 16 days of culture.(B) Integrated fluorescent intensities for TWIST1 with representative confocal images of human articular chondrocytes seeded in GelMA-based hydrogels channels after 16 days of culture.(C) Proposed mechanism of ACAN and COL2A synthesis upon thermomechanical loading through HIF1a signaling pathway. Schematic was created with BioRender.com.Error bars represent mean+standard deviation.

Fig 3: Chondroinduction response of HP-T stimulation on chondrocytesChondroinduction related gene (n = 6–9) and protein analyses (n = 3–6) for human adult chondrocyte-laden constructs subjected to different stimulations: “C” represents unstimulated control; “P” represents isolated HP stimulation; “T” represents isolated thermal stimulation; “PT” represents combined HP-T stimulation (as described in Figure 2). Relative mRNA expression of (A) SOX9 and TWIST1 as well as (B) their ratio. Immunofluorescent staining for (C) SOX9 (D) and TWIST1 protein (DAPI: blue; SOX9: yellow; TWIST1: magenta).(E) Relative gene expression for COL2A versus COL1A.(F) Relative gene expression of aggrecan (ACAN) and ADAMTS5.(G) Total GAG quantification normalized to the DNA quantity of the sample.(H) Heatmap of the LFQ (label-free quantification intensity from mass spectrometry, indicating protein abundance) intensity of various proteins identified by mass spectrometry of the human cartilage explant after 5 days of either free-swelling (“C”) or combined stimulation of HP and T (“PT”). Box and whisker plots represent the median, interquartile range, and the minimum and maximum values, with the “+” sign in the boxplots showing the mean. Bar charts show mean and standard error of the mean. Significance is correlated to the p value from Brown-Forsythe and Welch ANOVA tests or one-way ANOVA tests (see STAR Methods section for more details): p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗), p < 0.0001 (∗∗∗∗).