Novel hybrid scaffold for improving the wound repair process: evaluation of combined chitosan/eggshell/vitamin D scaffold for wound healing

The current design requirement of cutaneous wound healing is an ideal wound
dressing via biodegradable property through which fibroblasts can support, migrate
and proliferate. This study aimed to evaluate the effect of the synthesized hybrid
scaffold based on chitosan polymer and a natural source of calcium (Ca) and vitamin
D on wound healing. To achieve this purpose, three scaffolds include in chitosan
(CS), eggshell + chitosan (ES/CS) and chitosan + eggshell + vitamin D (CS/
ES/Vit D) were fabricated using the freeze-drying approach. Synthesized scaffolds
were characterized by field emission scanning electron microscopy (FESEM), Fourier
transformed infrared (FTIR), X-ray diffraction (XRD) and MTT assay. Histological
examinations of the wound healing were performed using hematoxylin–
eosin (H&E) and Masson’s trichrome staining methods on different days in the rat
full-thickness skin wound model. Finally, the effect of the synthesized scaffold was
evaluated on the TGF-β1 gene expression. The results of the scaffold characterization
showed that scaffolds have the homogeneity structure. The MTT assay indicated
that the cultured fibroblasts on the ES/CS and Vit D/ES/CS scaffold had viability
higher than those cultured on CS. Also, histological studies demonstrated an
increased effect on epithelialization and collagen production and accelerated wound
healing using designed scaffolds. The TGF-β1 gene expression in all scaffolds was