Journal: Materials Today Bio

Article Title: Injectable hypoxia-preconditioned cartilage progenitor cells-laden GelMA microspheres system for enhanced osteoarthritis treatment

doi: 10.1016/j.mtbio.2023.100637

Figure Lengend Snippet: Hypoxia-pretreated CGMs alleviate oxidative stress and inflammatory response and promote NRF2/HO-1 pathway activation in IL-1β-induced chondrocytes. (A) The level of malondialdehyde (MDA) in each group was detected after co-incubation for 48 ​h (n ​= ​3). (B) The levels of reactive oxygen species (ROS) in different groups were analyzed after co-incubation for 48 ​h (n ​= ​3). (C) Superoxide dismutase (SOD) activity levels were evaluated after different treatments after co-incubation for 48 ​h (n ​= ​3). (D) The concentrations of TNFα, IL6, and MMP13 were evaluated after co-incubation for 48 ​h by ELISA assay (n ​= ​3). (E–H) The mRNA expressions of the inflammatory factors ( Tnfα, Il6 , iNOS , and Cox2 ) in each group were detected after co-incubation for 48 ​h by PCR analysis (n ​= ​3). (I and J) The mRNA expressions of the antioxidant genes ( Sod1 and Sod2 ) were detected after co-incubation for 48 ​h by PCR analysis (n ​= ​3). (K) The mRNA expressions of the NRF2/HO-1 pathway-related genes ( Keap1 , Nrf2 , HO-1 , and NQO1 ) were detected after co-incubation for 48 ​h by PCR analysis (n ​= ​3). (L) The protein expressions of the NRF2/HO-1 pathway-related markers (KEAP1, NRF2, and HO-1) were assessed after co-incubation for 48 ​h by western blotting analysis. Data represent mean ​± ​S.D. ∗P ​< ​0.05, ∗∗P ​< ​0.01, ∗∗∗P ​< ​0.001 versus the Control group. # P ​< ​0.05, ## P ​< ​0.01, ### P ​< ​0.001 versus the IL-1β group. $ P ​< ​0.05, $$ P ​< ​0.01, $$$ P ​< ​0.001 versus the norm-CGMs group.

Article Snippet: The primary antibodies Collagen2 (Abcam, USA), Aggrecan (Abcam, USA), GDF5 (Abcam, USA), PRG4 (Abcam, USA), SOX9 (Preoteintech, USA), ADAMTS5 (Abcam, USA), MMP13 (Abcam, USA), TNFa (Abcam, USA), IL6 (Abcam, USA), SOD1 (Proteintech, USA), KEAP1 (Abcam, USA), NRF2 (Abcam, USA), HO-1 (Abcam, USA), β-actin (Abcam, USA), and horseradish peroxidase (HRP)-conjugated secondary antibody were used.

Techniques: Activation Assay, Incubation, Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Materials Today Bio

Article Title: Injectable hypoxia-preconditioned cartilage progenitor cells-laden GelMA microspheres system for enhanced osteoarthritis treatment

doi: 10.1016/j.mtbio.2023.100637

Figure Lengend Snippet: Hypoxia-pretreated CGMs ameliorate cartilage inflammatory response and oxidative stress and activate NRF2/HO-1 pathway in subacute OA rat. (A and B) Synovial inflammation in each group was determined by H&E staining and scored (n ​= ​5). (C) Immunohistochemical staining inflammatory factors (IL-1β, IL6, and TNFα) in different groups (n ​= ​5). Scale bar, 25 ​μm. (D–F) Quantification of IL-1β-positivity, IL6-positivity, and TNFα-positivity in chondrocytes after different treatments in subacute OA rat (n ​= ​5). (G) Immunohistochemical staining of antioxidant enzyme SOD1 and the NRF2/HO-1 pathway-associated markers (NRF2, HO-1, and NQO1) in different groups (n ​= ​5). Scale bar, 25 ​μm. (H) Quantification of SOD1-positive chondrocytes after different treatments (n ​= ​5). (I–K) Quantification of NRF2-positivity, HO-1-positivity, and NQO1-positivity in chondrocytes in different groups (n ​= ​5). Data represent mean ​± ​S.D. ∗∗∗P ​< ​0.001 versus the Sham group. # P ​< ​0.05, ### P ​< ​0.001 versus the PBS group. $$ P ​< ​0.01, $$$ P ​< ​0.001 versus the norm-CGMs group.

Article Snippet: The primary antibodies Collagen2 (Abcam, USA), Aggrecan (Abcam, USA), GDF5 (Abcam, USA), PRG4 (Abcam, USA), SOX9 (Preoteintech, USA), ADAMTS5 (Abcam, USA), MMP13 (Abcam, USA), TNFa (Abcam, USA), IL6 (Abcam, USA), SOD1 (Proteintech, USA), KEAP1 (Abcam, USA), NRF2 (Abcam, USA), HO-1 (Abcam, USA), β-actin (Abcam, USA), and horseradish peroxidase (HRP)-conjugated secondary antibody were used.

Techniques: Staining, Immunohistochemical staining

Journal: Materials Today Bio

Article Title: Injectable hypoxia-preconditioned cartilage progenitor cells-laden GelMA microspheres system for enhanced osteoarthritis treatment

doi: 10.1016/j.mtbio.2023.100637

Figure Lengend Snippet: Hypoxia-pretreated CGMs alleviate cartilage inflammation and oxidative stress and promote the activation of NRF2/HO-1 pathway in chronic OA rat. (A and B) Synovial inflammation in each group was determined by H&E staining and scored (n ​= ​5). (C) Immunohistochemical staining inflammatory factors (IL-1β, IL6, and TNFα) in articular cartilage (n ​= ​5). Scale bar, 25 ​μm. (D–F) Quantification of IL-1β-positivity, IL6-positivity, and TNFα-positivity in chondrocytes after different treatments in chronic OA rat (n ​= ​5). (G) Immunohistochemical staining of antioxidant enzyme SOD1 and the NRF2/HO-1 pathway-related markers (NRF2, HO-1, and NQO1) in different groups (n ​= ​5). Scale bar, 25 ​μm. (H) Quantification of SOD1-positive chondrocytes in each group (n ​= ​5). (I–K) Quantification of NRF2-positivity, HO-1-positivity, and NQO1-positivity in chondrocytes in different groups (n ​= ​5). Data represent mean ​± ​S.D. ∗∗P ​< ​0.01, ∗∗∗P ​< ​0.001 versus the Sham group. # P ​< ​0.05, ## P ​< ​0.01, ### P ​< ​0.001 versus the PBS group. $$ P ​< ​0.01 versus the norm-CGMs group.

Article Snippet: The primary antibodies Collagen2 (Abcam, USA), Aggrecan (Abcam, USA), GDF5 (Abcam, USA), PRG4 (Abcam, USA), SOX9 (Preoteintech, USA), ADAMTS5 (Abcam, USA), MMP13 (Abcam, USA), TNFa (Abcam, USA), IL6 (Abcam, USA), SOD1 (Proteintech, USA), KEAP1 (Abcam, USA), NRF2 (Abcam, USA), HO-1 (Abcam, USA), β-actin (Abcam, USA), and horseradish peroxidase (HRP)-conjugated secondary antibody were used.

Techniques: Activation Assay, Staining, Immunohistochemical staining