Multifaceted Characterization And In Vitro Assessment Of Polyurethane-Based Electrospun Fibrous Composite For Bone Tissue Engineering

Figures & data

Figure 1 FESEM images and EDX of (A, B) PE, (C, D) PE/WG and (E, F) PE/WG/TiO₂. Sample with size of 1 cm * 1 cm was cut and imaged in Hitachi SU8020 at different magnifications.

Figure 2 FTIR of PE (blue line), PE/WG (red line) and PE/WG/TiO₂ (green line). Sample with size of 1 cm * 1 cm was cut and measured in wavelength range between 600 and 4000 cm⁻¹ in Nicolet iS5.

Figure 3 XRD images of a) PE (black line), b) PE/WG (red line) and c) PE/WG/TiO₂ (blue line). Sample with size of 1 cm * 1 cm was cut and recorded in Rigaku having CuKα radiation in angle between 10° and 90°.

Table 1 Weight Loss Peaks Of The Electrospun Membranes

Peaks	PE	PE/WG	PE/WG/TiO2
First weight loss peak	210°C to 302°C	229°C to 352°C	230° C to 325°C
Second weight loss peak	302°C to 353°C	352°C to 447°C.	325°C to 355 °C
Third weight loss peak	353°C to 494°C	–	355 °C to 461 °C
Fourth weight loss peak	494°C to 760°C	–	–

Figure 4 (A) TGA and (B) Weight residue of PE (black line), PE/WG (red line) and PE/WG/TiO₂ (blue line). Sample weighing 3 mg was heated between temperature range of 30-1000°C under nitrogen atmosphere in Perkin-Elmer TGA unit.

Figure 5 Tensile strength of PE (black line), PE/WG (red line) and PE/WG/TiO₂ (blue line). Sample with size of 4 cm * 1 0.5 cm was stretched at a cross head speed of 10 mm/min with a 500 N load cell in Gotech Testing Machines, AI-3000.

Figure 6 AFM images of (A) PE, (B) PE/WG and (C) PE/WG/TiO_2. Sample with size of 1 cm * 1 cm was cut and scanned in 20 * 20 µm size with 256 * 256 pixels under normal atmosphere in Nanowizard, JPK instruments.

Figure 7 (A) APTT, (B) PT and (C) haemolytic assay of PE, PE/WG and PE/WG/TiO_2. For APTT assay sample with size of 1 cm * 1 cm was added with 50 µL of platelet-poor plasma (PPP) followed by incubating with 50 µL of reagent (rabbit brain cephaloplastin) and 50 µL CaCl₂ (0.025 M) to calculate the blood clotting time. For PT assay sample with size of 1 cm * 1 cm was added with 50 µL of platelet-poor plasma (PPP) followed by incubating with 50 µL of thromboplastin (Factor III) to calculate the blood clotting time. For haemolytic assay samples with size of 1 cm * 1 cm added to the mixture of citrated blood and diluted saline (4:5 v/v%) for 1 hr at 37°C. After this, the samples were centrifuged and optical density (OD) was measured at 542 nm.

Figure 8 SEM images of calcium deposition of (A) PE, (C) PE/WG and (E) PE/WG/TiO₂ and EDX graph showing calcium deposition of (B) PE, (D) PE/WG and (F) PE/WG/TiO_2. Samples (1 * 1 cm²) left in 1.5x SBF (pH 7.4; 37°C) for 14 days was imaged in Hitachi Tabletop TM3000.

Figure 9 MTS assay of PE, PE/WG and PE/WG/TiO_2. Samples with size of 0.5 cm * 0.5 cm were cut and placed in the 96 well plates. The scaffold was seeded with fibroblast cells with 10 × 10³ cells/cm² density and cultured for 5 days. After 5 days, the medium was added with 20% of MTS reagent for 4 h and optical density (OD) was measured at 490 nm.