Journal: Biomaterials Research

Article Title: Insulin-Like Growth Factor 2 Secreted from Mesenchymal Stem Cells with High Glutathione Levels Alleviates Osteoarthritis via Paracrine Rejuvenation of Senescent Chondrocytes

doi: 10.34133/bmr.0152

Figure Lengend Snippet: Intra-articular injection of primed MSCs suppresses OA progression without evidence of direct regeneration. (A) Representative histological analysis using Safranin-O and immunohistochemistry (IHC) (collagen type II, MMP13, collagen type X, DPP4, and human β2-microglobulin) in sham control and DMM-induced rabbits injected with PBS and naïve or primed MSCs. (B) Scoring of OA using OARSI grading system in sham control and DMM-induced OA rabbits that had undergone intra-articular injection of PBS and naïve or primed MSCs. (C to F) Quantification of collagen type II, MMP13, collagen type X, and DPP4 using scoring system. (G) Representative histological analysis using Safranin-O and DPP4 IHC staining in DMM-induced OA rabbits injected with PBS, primed MSCs, low-dose, mid-dose, or high-dose secretome, or triple high-dose secretome isolated from primed MSCs. (H and I) Scoring of OA using OARSI grading system and quantification of DPP4. ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Sections were then incubated with primary antibodies against collagen type II (Thermo Fisher Scientific, MA1-37493), MMP13 (R&D Systems, MAB511), collagen type X (Abcam, ab49945), DPP4 (Thermo Fisher Scientific, MA2607), and human β2 microglobulin (Abcam, ab759) diluted in 4% bovine serum albumin (BSA; Biosesang, AC1025-100-00) for 1 h at 37 °C.

Techniques: Injection, Immunohistochemistry, Control, Isolation

Journal: Biomaterials Research

Article Title: Insulin-Like Growth Factor 2 Secreted from Mesenchymal Stem Cells with High Glutathione Levels Alleviates Osteoarthritis via Paracrine Rejuvenation of Senescent Chondrocytes

doi: 10.34133/bmr.0152

Figure Lengend Snippet: Paracrine activity of primed MSC suppresses senescence phenotypes of OA chondrocytes. (A) Left: Representative immunofluorescence staining of p16 and p21 (red) in OA chondrocytes (OA), OA + naïve MSC media, OA + MSC priming media, and direct coculture of OA and naïve or primed MSCs. OA chondrocytes were distinguished with CellTracker Dil staining (green), and nuclei were counterstained with DAPI (blue). Right: Bar graphs showing quantification of p16 + OA chondrocytes (top) and quantification of p21 + OA chondrocytes ( n = 3 per group) (bottom). (B) Schematic illustration of the experiments. (C) mRNA levels of p16 and p21 in OA, OA + naïve CM, and OA + primed CM measured by RT-qPCR. (D) Representative SA-β-Gal staining and quantification for OA, OA + naïve CM, and OA + primed CM ( n = 3 per group). (E) Cell counts of OA, OA + naïve CM, and OA + primed CM for 14 d to evaluate proliferation ( n = 3 per group). (F) mRNA expression of matrix degradation factors (ADAMTS5 and MMP13) in OA, OA + naïve CM, and OA + primed CM ( n = 3 per group). (G) Schematic illustration of indirect coculture system using transwell. (H) mRNA levels of p16 and p21 in OA, OA + naïve MSCs, and OA + primed MSCs measured by RT-qPCR. (I) Representative SA-β-Gal staining and quantification for OA, OA + naïve MSCs, and OA + primed MSCs ( n = 3 per group). (J) Cell counts of OA, OA + naïve MSCs, and OA + primed MSCs for 14 d to evaluate proliferation ( n = 3 per group). (K) mRNA expression of matrix degradation factors (ADAMTS5 and MMP13) in OA, OA + naïve MSCs, and OA + primed MSCs ( n = 3 per group). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Sections were then incubated with primary antibodies against collagen type II (Thermo Fisher Scientific, MA1-37493), MMP13 (R&D Systems, MAB511), collagen type X (Abcam, ab49945), DPP4 (Thermo Fisher Scientific, MA2607), and human β2 microglobulin (Abcam, ab759) diluted in 4% bovine serum albumin (BSA; Biosesang, AC1025-100-00) for 1 h at 37 °C.

Techniques: Activity Assay, Immunofluorescence, Staining, Quantitative RT-PCR, Expressing

Journal: Biomaterials Research

Article Title: Insulin-Like Growth Factor 2 Secreted from Mesenchymal Stem Cells with High Glutathione Levels Alleviates Osteoarthritis via Paracrine Rejuvenation of Senescent Chondrocytes

doi: 10.34133/bmr.0152

Figure Lengend Snippet: Knockdown of IGF2 expression in primed MSCs using siRNA inhibits therapeutic efficacy in vivo. (A) Representative histological analysis using Safranin-O and IHC (collagen type II, MMP13, collagen type X, and DPP4) in sham control and DMM-induced rabbits injected with PBS, primed MSCs, or primed MSCs with IGF2 knockdown (IGF2KD MSCs). (B) Scoring of OA using OARSI grading system in sham control and DMM-induced OA rabbits that had undergone intra-articular injection of PBS, primed MSCs, or IGF2KD MSCs. (C to F) Quantification of collagen type II, MMP13, collagen type X, and DPP4 using scoring system. ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Sections were then incubated with primary antibodies against collagen type II (Thermo Fisher Scientific, MA1-37493), MMP13 (R&D Systems, MAB511), collagen type X (Abcam, ab49945), DPP4 (Thermo Fisher Scientific, MA2607), and human β2 microglobulin (Abcam, ab759) diluted in 4% bovine serum albumin (BSA; Biosesang, AC1025-100-00) for 1 h at 37 °C.

Techniques: Knockdown, Expressing, Drug discovery, In Vivo, Control, Injection

Journal: Journal of translational medicine

Article Title: Pulsed electromagnetic fields potentiate bone marrow mesenchymal stem cell chondrogenesis by regulating the Wnt/β-catenin signaling pathway.

doi: 10.1186/s12967-024-05470-7

Figure Lengend Snippet: Fig. 4 3 mT PEMFs promoted chondrogenic differentiation of BMSCs in early stage of chondrogenesis. (A) Representative immunofluorescence images of ACAN (red), COL2A (green), SOX9 (green) and DAPI (blue) in both 0 mT and 3 mT groups of day 7 pellets. (B) Quantification of the fluo rescence intensities for ACAN, COL2A and SOX9 of day 7 pellets. (C) Quan tification of the transcript levels of acan, col2a, sox9, mmp3 and mmp13 of day 7 pellets. (D) Representative immunofluorescence images of ACAN (red), COL2A (green), SOX9 (green) and DAPI (blue) in both 0 mT and 3 mT groups of day 14 pellets. (E) Quantification of the fluorescence intensi ties for ACAN, COL2A and SOX9 of day 14 pellets. (F) Quantification of the transcript levels of acan, col2a, sox9, mmp3 and mmp13 of day 14 pellets. ns, p>0.05; * p<0.05; ** p<0.01; *** p<0.001. Student’s t test and one-way ANOVA were used for comparison between two groups and multiple groups, respectively. Scale bar: 100 mm

Article Snippet: The commercial antibodies used included ACAN (Novus Cat# NB600-504), COL2A (Proteintech Cat# 28459-1-AP), SOX9 (Huabio Cat# ET1611-56), MMP3 (Huabio Cat# ET1705-98), MMP13 (Novus Cat# NBP2-45887), p65 (Cell Signaling Technology Cat# 8242), pp65 (Cell Signaling Technology Cat# 3033), STAT3 (Cell Signaling Technology Cat# 4904), p-STAT3 (Cell Signaling Technology Cat# 9145), β-catenin (Huabio Cat# ET1601-5), sFRP3 (Santa Cruz Cat# sc-514,350), β-actin (Huabio Cat# M1210-2), HRPconjugated goat anti-rabbit IgG (Huabio Cat# HA1001), and HRP-conjugated goat anti-mouse IgG (Huabio Cat# HA1006).

Techniques: Immunofluorescence, Fluorescence, Comparison

Journal: Journal of translational medicine

Article Title: Pulsed electromagnetic fields potentiate bone marrow mesenchymal stem cell chondrogenesis by regulating the Wnt/β-catenin signaling pathway.

doi: 10.1186/s12967-024-05470-7

Figure Lengend Snippet: Fig. 6 3 mT PEMFs inhibited inflammation and promoted chondrogenic differentiation of BMSCs under inflammation condition. (A) For day 7 pellets: the protein levels of p65, p-p65, STAT3 and p-STAT3, which showed the level of inflammation, were investigated by western blotting. In addition, the catabolic-related proteins of chondrocytes (MMP3 and MMP13) were also displayed. Quantification of relative expression of these proteins were per formed. (B) For day 14 pellets: the protein levels of the p65, p-p65, STAT3, p-STAT3, MMP3 and MMP13 and quantification of relative expression of these proteins. ns, p>0.05; * p<0.05; ** p<0.01; *** p<0.001. Student’s t test and one-way ANOVA were used for comparison between two groups and multiple groups, respectively

Article Snippet: The commercial antibodies used included ACAN (Novus Cat# NB600-504), COL2A (Proteintech Cat# 28459-1-AP), SOX9 (Huabio Cat# ET1611-56), MMP3 (Huabio Cat# ET1705-98), MMP13 (Novus Cat# NBP2-45887), p65 (Cell Signaling Technology Cat# 8242), pp65 (Cell Signaling Technology Cat# 3033), STAT3 (Cell Signaling Technology Cat# 4904), p-STAT3 (Cell Signaling Technology Cat# 9145), β-catenin (Huabio Cat# ET1601-5), sFRP3 (Santa Cruz Cat# sc-514,350), β-actin (Huabio Cat# M1210-2), HRPconjugated goat anti-rabbit IgG (Huabio Cat# HA1001), and HRP-conjugated goat anti-mouse IgG (Huabio Cat# HA1006).

Techniques: Western Blot, Expressing, Comparison

Journal: Journal of translational medicine

Article Title: Pulsed electromagnetic fields potentiate bone marrow mesenchymal stem cell chondrogenesis by regulating the Wnt/β-catenin signaling pathway.

doi: 10.1186/s12967-024-05470-7

Figure Lengend Snippet: Fig. 9 PEMFs exerted cartilage protection effect in DMM induced OA model and inhibited Wnt-βcatenin signaling pathway in vivo. (A) Schematic of the in vivo group set. (B) Representative Toluidine Blue-stained joint sections from mice post sham-DMM surgery, DMM surgery, DMM surgery and PEMFs treatment and quantification of OARSI scores. Scale bar: 100 mm. (C) Representative immunohistochemical staining of ACAN, COL2A, SOX9, MMP13, ADAMTS4, Wnt3a, β-catenin and IL-1β (n = 3). Scale bar: 100 mm

Article Snippet: The commercial antibodies used included ACAN (Novus Cat# NB600-504), COL2A (Proteintech Cat# 28459-1-AP), SOX9 (Huabio Cat# ET1611-56), MMP3 (Huabio Cat# ET1705-98), MMP13 (Novus Cat# NBP2-45887), p65 (Cell Signaling Technology Cat# 8242), pp65 (Cell Signaling Technology Cat# 3033), STAT3 (Cell Signaling Technology Cat# 4904), p-STAT3 (Cell Signaling Technology Cat# 9145), β-catenin (Huabio Cat# ET1601-5), sFRP3 (Santa Cruz Cat# sc-514,350), β-actin (Huabio Cat# M1210-2), HRPconjugated goat anti-rabbit IgG (Huabio Cat# HA1001), and HRP-conjugated goat anti-mouse IgG (Huabio Cat# HA1006).

Techniques: In Vivo, Staining, Immunohistochemical staining