Journal: Bioactive Materials

Article Title: Artificial mitochondria ameliorates osteoarthritis through restoring cellular energy metabolism homeostasis

doi: 10.1016/j.bioactmat.2026.02.028

Figure Lengend Snippet: The upregulation of CK in the cartilage and meniscus during early OA. (a) The contents of CK in the synovial fluid, articular cartilage and meniscus tissue of OA patients are measured with an ELISA kit. n = 20, one-way ANOVA, ∗∗∗∗ represents p < 0.0001. (b, c) Immunofluorescence staining (b) and semi-quantitative analysis (c) of CK levels within synovial fluid, articular cartilage and meniscus tissues of OA patients. CK represents creatine kinase. n = 6, one-way ANOVA, ∗∗ represents p < 0.01 and ∗∗∗∗ represents p < 0.0001. White scale bar: 50 μm; yellow scale bar: 10 μm. (d, e) Immunofluorescence staining (d) and semi-quantitative analysis (e) of CK levels in the knee joints of normal, early OA and late OA mice. F represents the femur, T represents the tibia, M represents the meniscus, AC represents the articular cartilage and SB represents the subchondral bone. n = 6, one-way ANOVA, ∗∗∗∗ represents p < 0.0001. White scale bar: 50 μm; yellow scale bar: 10 μm. (f – h) SDS-PAGE (f) results and semi-quantitative analysis of CK (g) and MMP13 (h) levels in rat chondrocytes after in vitro LPS stimulation for different durations. n = 3, one-way ANOVA, ∗ represents p < 0.05, ∗∗ represents p < 0.01, ∗∗∗ represents p < 0.0005, ∗∗∗∗ represents p < 0.0001 and ns represents no significant difference. MMP13 represents matrix metalloproteinase-13, and CK represents creatine kinase. (i – k) SDS‒PAGE results (i) and semi-quantitative analysis of CK (j) and MMP13 (k) levels in rat meniscus fibrochondrocytes after in vitro LPS stimulation for different durations. n = 3, one-way ANOVA, ∗∗ represents p < 0.01, ∗∗∗ represents p < 0.0005, ∗∗∗∗ represents p < 0.0001 and ns represents no significant difference.

Article Snippet: The rabbit anti MMP13 antibody was purchased from Proteintech (18165-1-AP).

Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, SDS Page, In Vitro

Journal: Bioactive Materials

Article Title: Collagen II hydrogel-mediated sustained delivery of lacosamide attenuates cartilage degeneration and pain in osteoarthritis

doi: 10.1016/j.bioactmat.2026.02.045

Figure Lengend Snippet: Non-selective Nav1.7 inhibitors regulate chondrocyte metabolism . (A) mRNA levels of COL2 and ACAN in primary human OA chondrocytes treated with 10 μM LCM, CBZ, or OXC for 24h. (B) mRNA levels of MMP13 and ADAMTS5 in primary human OA chondrocytes stimulated with 20 ng/mL IL-1β and 10 μM LCM, CBZ or OXC for 24h. (C, D) COL2 and ACAN (C) , MMP13 and ADAMTS5 (D) mRNA expression in human OA chondrocytes treated with various doses of LCM with/without IL-1β stimulated conditions. (E, F) mRNA levels of anabolic markers Col2 and Acan , as well as catabolic enzymes Mmp13 and Adamts5 , in primary chondrocytes isolated from WT and chondrocyte specific Nav1.7 knockout mice under basal conditions or following stimulation with 20 ng/mL IL-1β, with or without 10 μM LCM co-treatment for 24 h. (G) Representative immunofluorescence staining showing COL2 (green) and MMP13 (green) expression in C28/I2 chondrocytes treated with 20 ng/ml IL-1β and 10 nM LCM for 48h. Nuclei were counterstained with DAPI (blue). Scale bar = 50 μm. (H) Quantification of average optical density (AOD) shown in G. (I) Safranin O/Fast Green and IHC staining of human OA cartilage explants cultured with 20 ng/ml IL-1β and 10 nM LCM for 5 days. Scale bar = 100 μm. (J) Quantification of GAG area and OARSI scores in I. (K) Quantification of IHC staining in I. Data are presented as mean ± SD. Statistical analysis: one-way ANOVA with post hoc comparison (A-F and H, J, K).

Article Snippet: Cells were fixed with ice-cold methanol for 5 min and then blocked with 5% bovine serum albumin for 1 h. Samples were subsequently incubated with primary antibodies against COL2 (1:200, ProteinTech, 28459-1-AP) or MMP13 (1:200, ProteinTech, 18165-1-AP) overnight at 4 °C, followed by incubation with fluorophore-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Isolation, Knock-Out, Immunofluorescence, Staining, Immunohistochemistry, Cell Culture, Comparison

Journal: Bioactive Materials

Article Title: Collagen II hydrogel-mediated sustained delivery of lacosamide attenuates cartilage degeneration and pain in osteoarthritis

doi: 10.1016/j.bioactmat.2026.02.045

Figure Lengend Snippet: LCM regulated chondrocyte metabolism through enhancing HSP70 and midkine secretion . (A) ELISA quantification of midkine and HSP70 levels in conditioned medium (CM) collected from human OA chondrocytes treated with 10 nM LCM for 48 h. (B) mRNA levels of COL2 and ACAN in chondrocytes cultured with LCM- CM for 24 h. (C) mRNA levels of MMP13 and ADAMTS5 in human OA chondrocytes treated with 20 ng/mL IL-1β and LCM-CM for 24 h. (D) mRNA expression of COL2 and ACAN in chondrocytes treated with LCM-CM in the presence or absence of anti-HSP70 (αHSP70) antibody for 24 h. (E) mRNA expression of MMP13 and ADAMTS5 in human OA chondrocytes stimulated with 20 ng/ml IL-1β and LCM-CM in the presence or absence of anti-midkine (αMDK) antibody for 24 h. (F, G) Representative immunofluorescence staining of MMP13 (F) and COL2 (G) in C28/I2 chondrocytes with/without IL-1β, LCM-CM, anti-HSP70 antibody and anti-midkine antibody. Nuclei were counterstained with DAPI (blue). Scale bar = 50 μm. (H) Quantification of average optical density (AOD) of MMP13 and COL2 immunofluorescence shown in F and G. (I) Safranin O/Fast Green and IHC staining for MMP13 in human OA cartilage explants cultured with 20 ng/mL IL-1β in the presence or absence of LCM-CM or anti-midkine antibody for 5 days. Scale bar = 100 μm. (J) Safranin O/Fast Green and IHC staining for COL2 in cartilage explants cultured with LCM-CM with or without anti-HSP70 antibody for 5 days. Scale bar = 100 μm. (K) Quantification of GAG area and OARSI scores in I. (L) Quantification of IHC staining in I. (M) Quantification of GAG area and OARSI scores in J. (N) Quantification of IHC staining in J. Data are presented as mean ± SD. Statistical analysis: unpaired two-tailed Student's t-test (A, B) and one-way ANOVA with post hoc comparison (C-E and H, K-N).

Article Snippet: Cells were fixed with ice-cold methanol for 5 min and then blocked with 5% bovine serum albumin for 1 h. Samples were subsequently incubated with primary antibodies against COL2 (1:200, ProteinTech, 28459-1-AP) or MMP13 (1:200, ProteinTech, 18165-1-AP) overnight at 4 °C, followed by incubation with fluorophore-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Immunofluorescence, Staining, Immunohistochemistry, Two Tailed Test, Comparison

Journal: Bioactive Materials

Article Title: Collagen II hydrogel-mediated sustained delivery of lacosamide attenuates cartilage degeneration and pain in osteoarthritis

doi: 10.1016/j.bioactmat.2026.02.045

Figure Lengend Snippet: Systemic administration of LCM attenuates OA progression and pain . (A) Schematic of the experimental outline. Twelve-week-old male WT mice underwent DMM surgery. Beginning 4 weeks post-surgery, mice were treated daily with either CBZ (10 mg/kg) or LCM (1, 10, or 50 mg/kg) by oral gavage for a total duration of 8 weeks. (n = 6). (B) Representative Safranin O/Fast Green stained images of knee joints at 12 weeks post-DMM surgery, systemically treated with CBZ (10 mg/kg/day) or LCM (1, 10, or 50 mg/kg/day) (n = 6). Scale bar = 100 μm. (C) Quantification of OARSI scores, subchondral bone plate (SBP) thickness, osteophyte formation, and synovitis. (D, E) Behavioral outcomes assessed by open-field travel distance (D) and von Frey test (E). (F) Representative IHC staining of Col2, Aggrecan neoepitope, Mmp13, and Adamts5 in joint sections (n = 6). Scale bar = 50 μm ∗Vehicle versus LCM (50 mg/kg/day), # Vehicle versus LCM (10 mg/kg/day),(∗ ,# P < 0.05; ∗∗ ,## P < 0.01). Data are presented as mean ± SD. Statistical analysis: one-way ANOVA with post hoc comparison (C) and unpaired two-tailed Student's t-test (D, E).

Article Snippet: Cells were fixed with ice-cold methanol for 5 min and then blocked with 5% bovine serum albumin for 1 h. Samples were subsequently incubated with primary antibodies against COL2 (1:200, ProteinTech, 28459-1-AP) or MMP13 (1:200, ProteinTech, 18165-1-AP) overnight at 4 °C, followed by incubation with fluorophore-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Staining, Immunohistochemistry, Comparison, Two Tailed Test

Journal: Bioactive Materials

Article Title: Collagen II hydrogel-mediated sustained delivery of lacosamide attenuates cartilage degeneration and pain in osteoarthritis

doi: 10.1016/j.bioactmat.2026.02.045

Figure Lengend Snippet: Intra-articular delivery of LCM protects against OA . (A) Schematic of the experimental outline. Twelve-week-old male WT mice underwent DMM surgery. Beginning 4 weeks post-surgery, mice received intra-articular injections of CBZ (1 mg/kg, three times per week) or LCM (0.1 or 1 mg/kg, three times per week) for a total duration of 8 weeks (n = 6). (B) Representative Safranin O/Fast Green stained sections of knee joints at 12 weeks post-surgery, treated with or without CBZ (1 mg/kg, 3 × /week) or LCM (0.1 or 1 mg/kg, 3 × /week) intra-articularly (n = 6). Scale bar = 100 μm. (C) Quantification of OARSI scores, subchondral bone plate (SBP) thickness, osteophyte formation, and synovitis. (D, E) Behavioral outcomes assessed by open-field travel distance (D) and von Frey test (E). (F) Representative IHC staining of Col2, Aggrecan neoepitope, Mmp13, and Adamts5 in joint sections (n = 6). Scale bar = 50 μm ∗Vehicle versus LCM (1 mg/kg, 3 × /week), # Vehicle versus LCM (0.1 mg/kg, 3 × /week),(∗ ,# P < 0.05). Data are presented as mean ± SD. Statistical analysis: one-way ANOVA with post hoc comparison (C) and unpaired two-tailed Student's t-test (D, E).

Article Snippet: Cells were fixed with ice-cold methanol for 5 min and then blocked with 5% bovine serum albumin for 1 h. Samples were subsequently incubated with primary antibodies against COL2 (1:200, ProteinTech, 28459-1-AP) or MMP13 (1:200, ProteinTech, 18165-1-AP) overnight at 4 °C, followed by incubation with fluorophore-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Staining, Immunohistochemistry, Comparison, Two Tailed Test

Journal: Bioactive Materials

Article Title: Collagen II hydrogel-mediated sustained delivery of lacosamide attenuates cartilage degeneration and pain in osteoarthritis

doi: 10.1016/j.bioactmat.2026.02.045

Figure Lengend Snippet: Hydrogel-mediated delivery of LCM provides prolonged joint retention and therapeutic efficacy . (A) Schematic of the experimental outline. Twelve-week-old male WT mice underwent DMM surgery. Beginning at 4 weeks post-surgery, mice received intra-articular injections of either blank hydrogel or LCM-loaded hydrogel once monthly for 2 months (n = 6). (B) Safranin O/Fast Green–stained sections of knee joints 12 weeks post-DMM surgery, following intra-articular injection of LCM-loaded hydrogel (n = 6). Scale bar = 100 μm. (C) Quantification of OARSI scores, SBP thickness, osteophyte size, and synovitis in each group. (D, E) Behavior test outcomes assessed by open-field movement (D) and von Frey test (E). (F) Representative IHC staining of Col2, Aggrecan neoepitope, Mmp13, and Adamts5 in joint sections (n = 6). Scale bar = 50 μm. (G) Quantification of IHC-positive staining in (F). ∗Vehicle versus LCM-loaded hydrogel, (∗ P < 0.05; ∗∗ P < 0.01, ∗∗∗ P < 0.001). Data are presented as mean ± SD. Statistical analysis: one-way ANOVA with post hoc comparison (C, G) and unpaired two-tailed Student's t-test (D, E).

Article Snippet: Cells were fixed with ice-cold methanol for 5 min and then blocked with 5% bovine serum albumin for 1 h. Samples were subsequently incubated with primary antibodies against COL2 (1:200, ProteinTech, 28459-1-AP) or MMP13 (1:200, ProteinTech, 18165-1-AP) overnight at 4 °C, followed by incubation with fluorophore-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Drug discovery, Staining, Injection, Immunohistochemistry, Comparison, Two Tailed Test