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. 2021 Sep 27;9:716610. doi: 10.3389/fcell.2021.716610

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Copyright © 2021 Feng, Liu, Ding, Qiu and Chen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Sonic hedgehog (SHH) induced chondrocyte apoptosis and gene expression of the markers of hedgehog (HH) signaling, catabolism, senescence, and senescence-associated secretory phenotype (SASP) in chondrocytes. (A) Micrographs of primary osteoarthritic chondrocytes (OACs) and their treatment with human SHH recombinant protein for 24 and 48 h. SHH treatment induced shrunken cell shape and cell detachment. Images are representative of cell culture samples in triplicates. Scale bar = 125 μm. (B) Cell number change of OAC in response to human recombinant SHH incubation for 24 and 48 h. n = 4 individual biological samples. (C) Cell proliferation rate of OAC in response to human recombinant SHH incubation for 24 and 48 h. Cell proliferation rate was quantified by the CCK-8 proliferation assay. n = 5 biological replicates. (D) Fluorescence cytochemical analysis of chondrocyte apoptosis induced by human recombinant SHH. Live chondrocytes were stained blue with CytoCalcein Violet 450, while apoptotic cells were stained green with Apopxin green indicator. Scale bars = 125 μm. (E) The percentage of OAC apoptosis induced by human recombinant SHH. n = 4 individual biological samples. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of transcripts encoding markers of HH signaling (F) chondrogenesis (G), catabolism (H), and cell senescence and SASP (I) in primary OAC. Ctrl, Control (mock treatment).