Rise magna

Fixed rat RC-humeral complex specimens were placed in decalcifying fluid for 1 month until they were suitable for sectioning. After dehydration, embedding, sectioning (5 μm), and staining with SO/Fast Green, the tendon-to-bone interface was observed under a light microscope. For XRM, after fixation using 4% polyformaldehyde, the RC–humeral complex was immersed in Lugol's iodine solution for 48 h. Subsequently, the complex was transferred to 70% ethanol for 12 h and stored in 100% ethanol. Then, the 3-dimensional microstructure of the tendon-to-bone interface was observed using a Zeiss 520 Versa microscope, with exposure for 3 s at 7 W and 80 kV. Dragonfly (Object Research Systems, Canada) was used for remodeling. For SEM evaluation, after frozen sectioning, the sections were observed using RISE-MAGNA (TESCAN, Czech Republic). For immunofluorescence staining, sections were stained with the corresponding antibody and observed under an inverted fluorescence microscope. After the harvest of the rat RC–humeral complex, 1 mm3 tendon tissue was collected and fixed using 2.5% glutaraldehyde. After dehydration and embedding in resin, the tissue was cut into thin slices (5 μm) and observed using a Tecnai G2 microscope (Thermofisher, USA).

To observe the morphology of the cytoskeleton, the LMPCCs were immersed in 100 mM sucrose solution until they sank to the bottom and then washed with water for several times. The black-violet precipitates in the lower layer were collected with a pipette and transferred to a clean silicon wafer or a mica sheet. After drying at room temperature, the samples were observed with a transmission electron microscope (RISE-MAGNA, TESCAN, Czech Republic) and AFM (MFP-3D, Oxford Instrument, UK).

A Raman system (TESCAN RISE-MAGNA, Czech) was used to measure the Raman spectrum of the SWCNT film. A 532 nm laser was used as the excitation light whose spot size was 0.46 μm under 100× objective. A laser power of 0.507 mW, a numerical aperture of 0.75, and an exposure time of 15 s per spot were used in the Raman measurements.