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. 2023 Mar 29;27:98–112. doi: 10.1016/j.bioactmat.2023.03.017

Fig. 1.

Fig. 1

Hypoxia preconditioning enhanced the paracrine functions of human BM-MSC in vitro. (A) The proliferation of MSCs and chondrocytes were analyzed by quantitation of DNA amounts after 24, 48, and 72 h treated with NCM, HCM-5%, and HCM-1%, presented as ratio to 24 h NC group. (B) H&E staining of migrated MSCs and chondrocytes, and quantitative analysis of cell migration, presented as ratio to NC group. Scale bar = 200 μm. (C) Safranin O and COL-2 immunohistochemical staining, and (D) quantitative analysis of sGAG and COL-2 deposition, in chondrocyte pellets after 7 days in non-inflammation condition. Scale bar = 200 μm. (E) Safranin O staining and immunohistochemical staining, and quantitative analysis of COL-2, COL-1, ADAMTS5 content, of chondrocyte pellets after 7 days in IL-1β induced inflammation. Scale bar = 200 μm. (F) Quantitative analysis of sGAG and COL-2 deposition, and (G) transcription of IL-1β and IL-6 genes in chondrocyte pellets after 7 days in IL-1β induced inflammation. Data is presented as ratio to NC group. (H) SA-beta-Gal staining of senescent chondrocytes (blue colour) and quantitative analysis of senescent cells, presented as ratio to NC group. Scale bar = 100 μm. All data represent the mean ± standard deviation, n = 4. ^ denotes p < 0.05 compared to the NC; @ denotes p < 0.05 compared to the PC; * denotes p < 0.05 compared to NCM. # denotes p < 0.05 compared to HCM-5%. (I) Transcription of IL-1β, TNF-α, and IL-6 genes in macrophages, presented as ratio to M1 macrophage. Data represent the mean ± standard deviation, n = 4. ^ denotes p < 0.05 compared to the M0; @ denotes p < 0.05 compared to the M1; * denotes p < 0.05 compared to NCM. # denotes p < 0.05 compared to HCM-5%.