Rheowin data manager software

Rheological properties of the TSH and the SV samples were evaluated with a Haake Mars Rheometer (ThermoFisher Scientific, Waltham, MA, USA), using a conic geometry (1°) of 35 mm diameter. The gap size was 52 µm and the volume of the sample analyzed was 200 µL. All analyses were performed using the RheoWin Data Manager software (version 4.82.0002, ThermoFisher Scientific, Waltham, MA, USA).
The viscosity of synthetic vitreous (SV) samples was measured by applying a shear rate from 0.01 to 300 s⁻¹ at 37 °C, and results were fitted with the Carreau-Yasuda mathematical model to obtain the absolute viscosity (η₀). An amplitude sweep from 10 to 600% was performed at constant parameters of 1 Hz and 37 °C to obtain the storage modulus G’ of the gels, as a mean of values on the linear-viscoelastic regime (LVR). To measure the gelation temperature (Tgel) of the TSH samples, a 20-step ramp in a range of 5–50 °C at a rate of 0.075 °C/sec was performed at a constant frequency f = 1 Hz and a constant shear stress τ = 1 Pa. G’ and G” were recorded at each step and the gelation temperature was obtained at the crossover.

PDMS substrates of varying rigidity was prepared based on the previous literature.^[12] The SYLGARD components A and B (Sylgard 184; Dow Corning, Midland, MI, USA) were mixed at 50:1 and 100:1 ratios and spread on cell culture vessels (10‐ or 15‐cm‐diameter dish, 48‐ or 6‐well plate) or coverslips (13 mm), and cured at 70 °C overnight. Sulfo‐SANPAH solution (0.1 mg mL⁻¹, 22 589; Thermo Scientific, Waltham, MA, USA) was dropped on the surface of the PDMS substrate, irradiated for 10 min with UV light, and irradiated again for 5 min after the removal of the Sulfo‐SANPAH solution. The PDMS substrate was then coated with collagen I (25 µg mL⁻¹) after washing twice with phosphate‐buffered saline (PBS). The mechanical properties of the PDMS substrates were characterized using a rotational rheometer (HAAKE MARS III; Thermo Scientific). Parallel plates were used in oscillation mode (0.1–10 Hz, 25 °C) and the results were analyzed using RheoWin Data Manager software (Thermo Scientific).

PDMS substrates with different levels of stiffness were prepared according to a previous study [27 (link)]. The components A and B of SYLGARD (Sylgard 184; Dow Corning, Midland, MI, USA) were mixed at ratios of 50:1 and 100:1. The mixture was then spread onto cell culture vessels (10- or 15 cm-diameter dish) and cured at 70 °C overnight. A solution of Sulfo-SANPAH (0.1 mg mL⁻¹, 22 589; Thermo Scientific, Waltham, MA, USA) was dropped onto the surface of the PDMS substrate and exposed to UV light for 10 min. After removing the Sulfo-SANPAH solution, it was further irradiated for 5 min. The PDMS substrate was then coated with collagen I (25 μg mL⁻¹) after being washed twice with phosphate-buffered saline (PBS). The mechanical properties of the PDMS substrates were evaluated using a rotational rheometer (HAAKE MARS III; Thermo Scientific) in oscillation mode (0.1–10 Hz, 25 °C) with parallel plates as described by a previous study[27 (link)]. The data obtained were analyzed using RheoWin Data Manager software (Thermo Scientific).