Scaffold, an essential element of tissue engineering, should provide proper physical and chemical properties and evolve suitable cell behavior for tissue regeneration. Polycaprolactone/Gelatin (PCL/Gel)-based nanocomposite scaffolds containing hydroxyapatite nanoparticles (nHA) and vitamin D3 (Vit D3) were fabricated using the electrospinning method. Structural and mechanical properties of the scaffold were determined by scanning electron microscopy (SEM) and tensile measurement. In this study, smooth and bead-free morphology with a uniform fiber diameter and optimal porosity level with appropriate pore size was observed for PCL/Gel/nHA nanocom-

Tissue engineering knowledge is a step toward the treatment of irreversible damages to human beings. In the present study, PCL/Gel, PCL/Gel/nHA, PCL/Gel/Vit D3 and PCL/Gel/nHA/Vit D3 (Polycaprolactone/Gelatin/Nanohydroxyapatite/Vitamin D3) composite scaffolds were successfully constructed using electrospinning method. The proliferation and differentiation of hADSCs into the bone phenotype were determined using MTT method, ALP activity, Von Kossa and Alizarin red staining, and qRT-PCR test. The simultaneous presence of nHA and vitamin D3 led to the increased activity of ALP in the early stages (on the 14 th day) and increased mineralization in the late stages (on the 21 th day) in differentiated hADSCs. Further, it was found that the use of nHA and vitamin D3 resulted in increased expression of BGLAP and COLL I and reduced expression of ALP and RUNX2 in hADSCs for 21 days. The results indicated that nHA and vitamin D3 have a synergistic effect on the osteogenic differentiation of hADSCs.

Electrospinning is considered a powerful method for the production of fibers in the nanoscale size. Small pore size results in poor cell infiltration, cell migration inhibition into scaffold pores and low oxygen diffusion. Electrospun polycaprolactone/gelatin/nano-hydroxyapatite (PCL/Gel/nHA) scaffolds were deposited into two types of fiber collectors (novel rotating disc and plate) to study fiber morphology, chemical, mechanical, hydrophilic, and biodegradation properties between each other. The proliferation and differentiation of MG-63 cells into the bone phenotype were determined using MTT method, alizarin red staining and alkaline phosphatase (ALP) activity. The rates for disc rotation were 50 and 100 rpm. The pore size measurement results indicated that the fibers produced by the disc rotation collector with speed rate 50 rpm have larger pores as compared to fibers produced by disc rotation at 100 rpm and flat plate collectors. A randomly structure with controlled pore size (38.65 ±0.33 μm) and lower fiber density, as compared to fibers collected by disc rotation with speed rate 100 rpm and flat plate collectors, was obtained. Fibers collected on the rotating disc with speed rate 50 rpm, were more hydrophilic due to larger pore size and therefore, faster infiltration of water into the scaffold and the rate of degradation was higher. These results demonstrate that PCL/Gel/nHA scaffolds made through a rotating disc collector at 50 rpm are more feasible to be used in bone tissue engineering applications due to appropriate pore size and increased adhesion and proliferation of cells, ALP activity and mineral deposits.

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