After cell fixation, the samples were rinsed with PBS and then dehydrated with graded buy JNK-IN-8 concentrations of ethanol (20 vol.%, 30 vol.%, 40 vol.%, 50 vol.%, 70 vol.%, and 100 vol.% ethanol) for 10 min each. Finally, the samples were kept overnight in a vacuum oven and observed in FE-SEM to determine cell attachment. The samples for FE-SEM were coated by keeping the same conditions as described previously in the ‘Characterization’ section. However, the micrographs of each sample were taken at an accelerating voltage of 2 KV and with magnifications of 15 K. Results and discussions The three-way

stopcock connector was used as the solution blending tool before ejecting the solution into nanofibers. In this regard, Figure 3 demonstrates the degree of dispersion of HAp NPs in the silk solution. This optical micrograph was taken from silk/PEO and HAp/PEO composite solution immediately after mixing using the threeway connector. In this figure, we can clearly observe that HAp NPs are completely dispersed in the silk solution, which further confirms that HAp NPs can be easily carried along with the electrospinning solution during fiber formation. Electrospinning of silk solutions containing various amounts of HAp NPs (i.e., 0%, 10%, 30%, and 50%) afforded in the fabrication

of nanofibers with desirable Milciclib manufacturer morphology (Figure 4). Figure 4A represents the results RGFP966 concentration after electrospinning of pure silk solutions; it can be observed that nanofibers are smooth, uniform, continuous, and bead-free. Moreover, its counterparts containing HAp NPs are represented in Figure 4B,C,D. By observing these figures, one can come up with a simple conclusion that general morphology had not been affected by the addition of HAp NPs. However, it can be observed that there is a reasonable increase in fiber diameters due to the addition of HAp NPs. To find out the actual effect caused due to the addition of HAp NPs on nanofiber, the average diameters of nanofibers were calculated from randomly selected individual fibers (100 diameters measured per sample) using the image analyzer software (Innerview 2.0). In this regard, Figure 5 presents the bar graphs for diameters

calculated Dapagliflozin from each nanofiber combinations. It can be observed that pristine nanofibers had an average diameter of 110 ± 40 nm, and nanofibers modified with 10%, 30%, and 50% HAp NPs had increased diameters of 163 ± 45 nm, 273 ± 70 nm, and 212 ± 71 nm, which indicate the allocation of higher viscosity due to the presence of HAp NPs colloid which resulted in large droplet formation, giving it a tough bending instability during fiber formation and that finally resulted to the increase of the nanofiber diameters [26]. Figure 3 Optical micrograph of the composite solution containing silk/PEO and HAp/PEO after mixing using the threeway connector. Figure 4 Field emission scanning microscopy results. Of the pristine silk fibroin nanofibers (A), silk fibroin nanofibers modified with 10% HAp (B), 30% HAp (C), and 50% HAp (D).