recombinant klotho protein  (MedChemExpress)

Journal: Science Advances

Article Title: Bioactive fiber-reinforced hydrogel to tailor cell microenvironment for structural and functional regeneration of myotendinous junction

doi: 10.1126/sciadv.adm7164

Figure Lengend Snippet: ( A ) Schematic illustration of the fabrication of PLGA-reinforced SilMA hydrogel containing MSCs and Klotho (K/M@SM-PA). ( B ) SEM images of lyophilized PA, SM-PA, and K@SM-PA scaffolds. Low magnification: scale bars, 300 μm; high magnification: scale bars, 100 μm. ( C ) PLGA fiber diameter of PA, SM-PA, and K@SM-PA scaffolds ( n = 3 scaffolds per group). Beeswarm plots (small circular dots) represent individual fibers that are color coded in accordance with their corresponding scaffolds (squares). ( D ) Pore area of SilMA hydrogels in SM-PA and K@SM-PA scaffolds ( n = 3 scaffolds per group). Beeswarm plots (small circular dots) represent individual pores that are color coded in accordance with their corresponding scaffolds (squares). ( E and F ) Tensile modulus (E) and failure force (F) of PA, SM-PA, and K@SM-PA scaffolds, as well as native rat MTJ ( n = 5 MTJ samples or scaffolds per group). ( G ) Rheological strain sweeps from 0.01 to 100% strain at 10 rad/s. ( H ) Rheological frequency sweeps from 0.1 to 100 rad/s at 1% strain. ( I ) Water contact angles of PA, SM-PA, and K@SM-PA scaffolds captured at 1 s after water dropping. ( J ) Swelling ratio of PA, SM-PA, and K@SM-PA scaffolds at 6, 12, and 24 hours after incubation in PBS at 37°C ( n = 5 scaffolds per group). ( K ) Live/dead staining of MSCs cultured within the SM-PA and K@SM-PA hydrogels. Scale bars, 200 μm. ( L ) Cell viability of MSCs quantified from live/dead staining images ( n = 7 randomly selected microscopic images per group). ( M ) MSC proliferation measured by CCK-8 assay on days 1, 3, and 5 ( n = 6 independent experimental units per group). DMSO, 5% DMSO; Ctrl, normal growth medium. Results are shown as means ± SD, ns P ≥ 0.05, * P < 0.05, **** P < 0.0001. Schematic illustrations were created using BioRender ( www.biorender.com ).

Article Snippet: Recombinant Klotho protein (0.05 μg; 100-53, PeproTech, USA) and/or 2 × 10 6 primary rat bone marrow–derived MSCs were added to 50 μl of SilMA solution.

Techniques: Incubation, Staining, Cell Culture, CCK-8 Assay

Journal: Science Advances

Article Title: Bioactive fiber-reinforced hydrogel to tailor cell microenvironment for structural and functional regeneration of myotendinous junction

doi: 10.1126/sciadv.adm7164

Figure Lengend Snippet: ( A ) Schematic illustration of in situ implantation of the K/M@SM-PA hydrogel into the MTJ defect in rats. The PLGA scaffold was preimplanted into the defect area, and the SilMA hydrogel containing Klotho and MSCs was subsequently photocrosslinked in situ. ( B ) H&E staining of regenerated tissues in MTJ defects at 4 weeks after surgery. Black, blue, and red boxes indicate the repaired MTJ, tendon (T), and skeletal muscle (M), respectively. The black dashed lines show the interface of the muscle and tendon. Low magnification: scale bars, 100 μm; high magnification: scale bars, 25 μm. ( C ) Masson trichrome staining of regenerated tissues in MTJ defects at 4 weeks after surgery. Low magnification: scale bars, 100 μm; high magnification: scale bars, 25 μm. ( D to F ) IHC staining for Paxillin (MTJ marker), TNMD (tendon marker), and MYH4 (muscle marker) of regenerated tissues in MTJ defects at 4 weeks after surgery. Low magnification: scale bars, 200 μm; high magnification: scale bars, 40 μm. ( G ) Histology scores of regenerated tendon tissues evaluated from H&E staining images ( n = 3 rats per group). ( H ) Quantitative analysis of regenerated myofiber diameter from H&E staining images ( n = 3 rats per group). ( I to K ) Quantitative analysis of IHC staining for Paxillin, TNMD, and MYH4 of regenerated tissues at 4 weeks after surgery ( n = 4 randomly selected microscopic images per group). Results are shown as means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Schematic illustrations were created using BioRender ( www.biorender.com ).

Article Snippet: Recombinant Klotho protein (0.05 μg; 100-53, PeproTech, USA) and/or 2 × 10 6 primary rat bone marrow–derived MSCs were added to 50 μl of SilMA solution.

Techniques: In Situ, Staining, Immunohistochemistry, Marker