collagen type ii alpha 1 chain col2a1  (Santa Cruz Biotechnology)

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: MDSCs exhibit a dual role in the progression of osteoarthritis. (A) Overview of animal experiments, mice were randomly sorted into 3 groups (Sham, Control, MDSC, n = 6). The MDSC group received intra-articular injections of MDSCs twice a week, and the animals were sacrificed one and four weeks after surgery. (B) Micro-CT images of knee joint after injected MDSCs. (C) The relative joint space. (D–E) Safranin O-fast green, H&E, (F) The Mankin scores of knee joints. (G) COL2A1, and MMP-13 staining of knee joints. (H) Quantitative analysis of collagen II-positive cells and MMP-13 expression. (I) Volcano plot of differentially expressed genes (DEGs) between OA and Sham. (J) Heat map of differentially expressed genes between OA and sham. (K) Volcano plot of differentially expressed genes (DEGs) between OA and MDSC. (L) The gene set enrichment analysis (GSEA) of MDSC. (M) The kyoto encyclopedia of genes and genomes (KEGG) analysis of MDSC. (N) The expression levels of various IL-17 receptors between OA and MDSC. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Control, Micro-CT, Injection, Staining, Expressing

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: MDSCs exacerbate arthritis by promoting Th17 differentiation, but depleting MDSCs can alleviate the progression of osteoarthritis. (A) One week after the injection of MDSC, compared to the control group, the intra-articular levels of TNF-α, IFN-γ, IL-1β, and IL-6. (B) Flow cytometry showed the expression of IL-17A in the joints one week after the DMM surgery. One week after the injection of MDSC, compared to the control group, the intra-articular concentrations of IL-17 (C) in serum (D) in joint. (E) Immunofluorescence staining of CD4 and IL-17 in the joints and synovial membrane of mice one week after DMM surgery. (F) Immunofluorescence of COL2A1 and MMP-13 in ATDC5 cells co-cultured with Th17 cells for four days. (G) Quantitative analysis of immunofluorescence for COL2A1 and MMP-13. (H) Flow cytometry analysis of IL-17 expression and IL-17 levels in the culture supernatant after co-culturing MDSCs with CD4 + T cells for four days, compared to the control group. Mice were randomly sorted into 3 groups (Sham, Control, Anti-Gr-1, n = 6). The Anti-Gr-1 group received i.p. injected anti-Gr-1 antibodies (100 μg) twice a week, and animals were sacrificed four weeks after surgery. (I) Micro-CT images of knee joint after injection of anti-Gr-1 at 4 weeks. (J) Quantitative results of bone mineral density after injection of anti-Gr-1. (K) Safranin O-fast green, H&E staining of knee joints after injection of anti-Gr-1 at 4 weeks. (L) Total OARSI score in different groups after injection of anti-Gr-1. (T: tibial, F: femoral) (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Injection, Control, Flow Cytometry, Expressing, Immunofluorescence, Staining, Membrane, Cell Culture, Micro-CT

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: MDSCs accelerate cartilage regeneration by regulating macrophage M2 polarization. (A) Flow cytometry analysis of CD206 expression in joint. (B) Flow cytometry analysis of CD86 expression in joint. (C) One week after the injection of MDSCs, immunofluorescence of F4/80, CD86, and CD206 in the synovial membrane (n = 3). (D) Flow cytometriy analysis of CD206 expression after co-culturing MDSCs with RAW264.7 cells. (E) Cytokines levels of M2 macrophage-in the supernatant of the culture medium after co-culturing MDSCs with RAW264.7 cells. (F) Immunofluorescence staining of COL2A1 and MMP-13 after co-culturing M2 macrophages with ATDC5 cells for four days. (G) Quantitative analysis of immunofluorescence for COL2A1 and MMP-13. (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Flow Cytometry, Expressing, Injection, Immunofluorescence, Membrane, Staining

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: HAMA-MSN@TGF-β1/Anti-IL-1β microspheres accelerate cartilage repair by inhibiting Th17 differentiation. (A) MDSCs regulate the differentiation of CD4 + T cells and macrophages. (B) Flow cytometric analysis of IL-17 expression after co-culturing microspheres with CD4 + T cells for four days (n = 3). (C) Quantitative analysis of flow cytometry results and the concentration of IL-17 in the culture supernatant. After co-culturing MDSCs with CD4 T cells, collect the culture medium to treat ATDC5 cells and perform immunofluorescence staining of cartilage-related markers after co-culturing for four days with different groups of microspheres. (D) Immunofluorescence staining of COL2A1 (n = 6). (E) Immunofluorescence staining of MMP-13 (n = 6). (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Expressing, Flow Cytometry, Concentration Assay, Immunofluorescence, Staining

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: HAMA-MSN@TGF-β1/Anti-IL-1β microspheres accelerate cartilage repair by promoting M2 polarization of macrophages. (A) Flow cytometric analysis of CD206 expression after co-culturing microspheres with Raw264.7 cells. (B) After pre-treating ATDC5 cells with TNF-α for one day, co-culture them with M2 polarized macrophages for four days, followed by immunofluorescence staining of MMP-13. (C) Quantitative analysis of MMP-13 immunofluorescence (n = 6). (D) After pre-treating ATDC5 cells with TNF-α for one day, co-culture them with M2 polarized macrophages for four days, followed by immunofluorescence staining of COL2A1. (E) Quantitative analysis of COL2A1 immunofluorescence (n = 6). (F) The effect of HAMA-MSN@TGF-β1/Anti-IL-1β microspheres on the expression of genes related to cartilage metabolism (n = 3). (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Expressing, Co-Culture Assay, Immunofluorescence, Staining

Journal: Materials Today Bio

Article Title: Harnessing the dual immunomodulatory function of myeloid-derived suppressor cells to reshape the inflammatory microenvironment for osteoarthritis therapy

doi: 10.1016/j.mtbio.2025.102332

Figure Lengend Snippet: HAMA-MSN@TGF-β1/Anti-IL-1β alleviates cartilage loss. Mice were randomly sorted into 5 groups (Sham, Control, HAMA-MSN@TGF-β1, HAMA-MSN@Anti-IL-1β, HAMA-MSN@TGF-β1/Anti-IL-1β). (A) X-ray and Micro-CT images of the mice joints at week eight. (B) Quantitative analysis of joint space and subchondral bone density (n = 3). (C) Representative microscopic photographs of Safranine O/Fast Green and H&E staining. the areas in the black squares are magnified and shown separately. (red: cartilage proteoglycan; green: collagen of subchondral bone). (D) Total OARSI score in different groups. (E) Representative microscopic photographs of COL2A1 and MMP-13 immunohistochemical staining. (F, G) Quantitative analysis of MMP-13 and COL2A1 expression (n = 3). (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After rinsing the cells three times with PBS, collagen II alpha-1 chain (COL2A1) and matrix metalloproteinase 13 (MMP-13) antibodies (Boster, China) diluted at 1:100 were added and incubated overnight at 4 °C in a constant temperature shaker.

Techniques: Control, Micro-CT, Staining, Immunohistochemical staining, Expressing

Journal: Chinese Medicine

Article Title: Sodium tanshinone IIA sulfonate alleviates osteoarthritis through targeting SIRT1

doi: 10.1186/s13020-025-01166-2

Figure Lengend Snippet: Verification of Sirt1 cartilage-specific deletion in mice. A Schematic of TM administration. To induce Sirt1 deletion, TM (75 mg/kg) was intraperitoneally injected for 5 consecutive days into 8-week-old mice. The Sirt1 flox/flox and Col2a1-CreERT; Sirt1 flox/flox mice received the identical amounts of TM to ensure consistency. B PCR analysis of gene expression of Sirt1 and Col2a1-Cre in different tissues of mice. C RT-qPCR analysis of Sirt1 mRNA expression in cartilage of Control and Sirt1 cKO mice (n = 6/group). D Western blotting analysis of SIRT1 protein expression in cartilage of Control and Sirt1 cKO mice. β-actin was used as an internal control. Right: quantitative analysis of the protein expression of SIRT1 in cartilage of control and Sirt1 cKO mice (n = 3/group). Data are presented as mean ± SD, Student’s t test; ***P < 0.001

Article Snippet: The PVDF membrane was then subjected to blocking with 5% milk and subsequently incubated with the designated primary antibodies overnight at a temperature of 4 °C: β-actin (Proteintech, 66009-1-lg) (1:50,000), SIRT1 (Abcam, ab189494) (1:1000), NF-κB p65(CST, 8242) (1:10,000), NF-κB p65 (acetyl Lys310) (Abcam, ab19870) (1:10,000), Type II Collagen Alpha 1 Chain (COL2A1) (Proteintech, 28459-1-AP) (1:1000), Ferritin heavy chain 1 (FTH1) (Abcam, ab183781) (1:1000), Glutathione peroxidase 4 (GPX4) (Abcam, ab125066) (1:1000), acyl-CoA synthetase long-chain family member 4 (ACSL4) (Proteintech, 22401-1-AP) (1:5000), P300 (Santa Cruz, sc-48343)(1:1000).

Techniques: Injection, Gene Expression, Quantitative RT-PCR, Expressing, Control, Western Blot

Journal: Veterinary Medicine International

Article Title: Articular Cartilage Gene Expression after Coxofemoral Joint Luxation in the Dog

doi: 10.1155/2013/936317

Figure Lengend Snippet: Sequences of sense and antisense primers used for amplification in real-time PCR.

Article Snippet: Quantification of ten transcripts—aggrecan ( AGG ); type II collagen, alpha-1 chain ( COL2A1 ); matrix metalloproteinase-3 ( MMP-3 ); hyaluronan synthase-1 ( HAS-1 ); hyaluronan synthase-2 ( HAS-2 ); tissue inhibitor of metalloproteinase-1 ( TIMP-1 ); Bcl-2-associated X protein ( BAX ); B-cell lymphoma-2 ( BCL-2 ); cysteine aspartate-specific protease-3 ( CAS-3 ); and cysteine aspartate-specific protease-9 ( CAS-9 ) (Shanghai BlueGene Biotech, Shanghai, China)—was performed for all samples.

Techniques: Amplification, Sequencing

Journal: Veterinary Medicine International

Article Title: Articular Cartilage Gene Expression after Coxofemoral Joint Luxation in the Dog

doi: 10.1155/2013/936317

Figure Lengend Snippet: The relationship between relative expression of nonapoptotic genes and days of luxation ( AGG , aggrecan; COL2A , type II collagen (alpha-1 chain); HAS-1 , hyaluronan synthase-1; HAS-2 , hyaluronan synthase-2; TIMP , tissue inhibitor of metalloproteinase; MMP-3 , matrix metalloproteinase-3).

Article Snippet: Quantification of ten transcripts—aggrecan ( AGG ); type II collagen, alpha-1 chain ( COL2A1 ); matrix metalloproteinase-3 ( MMP-3 ); hyaluronan synthase-1 ( HAS-1 ); hyaluronan synthase-2 ( HAS-2 ); tissue inhibitor of metalloproteinase-1 ( TIMP-1 ); Bcl-2-associated X protein ( BAX ); B-cell lymphoma-2 ( BCL-2 ); cysteine aspartate-specific protease-3 ( CAS-3 ); and cysteine aspartate-specific protease-9 ( CAS-9 ) (Shanghai BlueGene Biotech, Shanghai, China)—was performed for all samples.

Techniques: Expressing

Journal: Veterinary Medicine International

Article Title: Articular Cartilage Gene Expression after Coxofemoral Joint Luxation in the Dog

doi: 10.1155/2013/936317

Figure Lengend Snippet: Linear regression equation R , R 2 , and significance level (Sig.) of candidate genes.

Article Snippet: Quantification of ten transcripts—aggrecan ( AGG ); type II collagen, alpha-1 chain ( COL2A1 ); matrix metalloproteinase-3 ( MMP-3 ); hyaluronan synthase-1 ( HAS-1 ); hyaluronan synthase-2 ( HAS-2 ); tissue inhibitor of metalloproteinase-1 ( TIMP-1 ); Bcl-2-associated X protein ( BAX ); B-cell lymphoma-2 ( BCL-2 ); cysteine aspartate-specific protease-3 ( CAS-3 ); and cysteine aspartate-specific protease-9 ( CAS-9 ) (Shanghai BlueGene Biotech, Shanghai, China)—was performed for all samples.

Techniques:

Journal: Nutrients

Article Title: Melatonin Prevents Chondrocyte Matrix Degradation in Rats with Experimentally Induced Osteoarthritis by Inhibiting Nuclear Factor-κB via SIRT1

doi: 10.3390/nu14193966

Figure Lengend Snippet: The circulating levels of melatonin (MT) and inflammatory factors in the serum of rats in each group. ( A – E ) Enzyme-linked immunosorbent assay (ELISA) kits were used to detect the circulating levels of MT, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), and tumor necrosis factor-alpha (TNF-α) in the serum of each group. All results represent mean ± standard deviation (SD) ( n = 3), * p < 0.05, ** p < 0.01 (compared to the model group). MT group: 30 mg/kg/2d MT.

Article Snippet: Lysate and RAPI were purchased from Biyuntian (Shanghai, China). nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p65), Phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-p65),alpha (IκBα), p-IκBα, TGF-β1, Smad2, iNOS, COX-2, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-13 (MMP-13), collagen type II alpha 1 chain (COL2A1), SIRT1, and ADAMTS-4 were purchased from ABclonal Technology (Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: Nutrients

Article Title: Melatonin Prevents Chondrocyte Matrix Degradation in Rats with Experimentally Induced Osteoarthritis by Inhibiting Nuclear Factor-κB via SIRT1

doi: 10.3390/nu14193966

Figure Lengend Snippet: Melatonin (MT) inhibited the nuclear factor κB (NF-κB) signaling and activated the transforming growth factor-beta 1 (TGF-β1)/SMAD family member 2 (Smad2) pathways in interleukin 1-beta (IL-1β)-treated chondrocytes. ( A , B ) Western blot (WB) was used to detect protein expression levels of p65, nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-p65), alpha (IκBα), p-IκBα, TGFβ1, and Smad2. * p < 0.05, ** p < 0.01 (compared to the model group).

Article Snippet: Lysate and RAPI were purchased from Biyuntian (Shanghai, China). nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p65), Phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-p65),alpha (IκBα), p-IκBα, TGF-β1, Smad2, iNOS, COX-2, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-13 (MMP-13), collagen type II alpha 1 chain (COL2A1), SIRT1, and ADAMTS-4 were purchased from ABclonal Technology (Wuhan, China).

Techniques: Western Blot, Expressing

Journal: Nutrients

Article Title: Melatonin Prevents Chondrocyte Matrix Degradation in Rats with Experimentally Induced Osteoarthritis by Inhibiting Nuclear Factor-κB via SIRT1

doi: 10.3390/nu14193966

Figure Lengend Snippet: Effects of EX527 on the nuclear factor kappa B (NF-κB) and transforming growth factor beta (TGF-β)/SMAD family member 2 (Smad2) pathways in melatonin (MT)-treated interleukin 1-beta (IL-1β)-treated chondrocytes. ( A , B ) Western blot (WB) was used to detect the protein expression levels of p65, nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor(p-p65), alpha (IκBα), p-IκBα, transforming growth factor beta (TGF-β1), and Smad family member 2 (Smad2). ( C , D ) Immunofluorescence (IF) staining was used to check the nuclear entry of p65 and Smad2 in each group. * p < 0.05, ** p < 0.01 (compared to the model group).

Article Snippet: Lysate and RAPI were purchased from Biyuntian (Shanghai, China). nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p65), Phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-p65),alpha (IκBα), p-IκBα, TGF-β1, Smad2, iNOS, COX-2, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-13 (MMP-13), collagen type II alpha 1 chain (COL2A1), SIRT1, and ADAMTS-4 were purchased from ABclonal Technology (Wuhan, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Staining

Journal: Nutrients

Article Title: Melatonin Prevents Chondrocyte Matrix Degradation in Rats with Experimentally Induced Osteoarthritis by Inhibiting Nuclear Factor-κB via SIRT1

doi: 10.3390/nu14193966

Figure Lengend Snippet: EX527 abrogated the inhibitory influence of MT on IL-1β-induced chondrocyte matrix degradation. ( A , C ) Western blot (WB) was used to detect the levels of matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-13 (MMP-13), ADAM metallopeptidase with thrombospondin type 1 motif 4 (ADAMTS-4), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and type II collagen. ( B , D ) Real-time PCR (qRT-PCR) was used to detect the levels of MMP-3, MMP-13, ADAMTS-4, COX-2, and type II collagen. ( E ) The intensity of collagen type II alpha 1 chain (COL2A1) in each group was detected by IF staining. ** p < 0.01 (compared to the model group).

Article Snippet: Lysate and RAPI were purchased from Biyuntian (Shanghai, China). nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p65), Phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor (p-p65),alpha (IκBα), p-IκBα, TGF-β1, Smad2, iNOS, COX-2, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-13 (MMP-13), collagen type II alpha 1 chain (COL2A1), SIRT1, and ADAMTS-4 were purchased from ABclonal Technology (Wuhan, China).

Techniques: Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Staining