A preliminary study on amelogenin-loaded electrospun scaffolds

Abstract

Amelogenin is a major enamel matrix protein onto which developing enamel forms. In the realm of tissue engineering, amelogenin has been studied and applied to periodontal and wound healing applications. This study introduces the first attempts of incorporating amelogenin within an electrospun scaffold. Amelogenin was extracted from porcine unerupted tooth buds and electrospun with poly(glycolic acid) and poly(ϵ-caprolactone). Protein release kinetics, mechanical properties, fiber diameter, mineralization potential, and cell adhesion properties of the amelogenin-blended scaffolds were studied and compared to the electrospun poly(glycolic acid) and poly(ϵ-caprolactone) controls. Electrospun scaffolds loaded with amelogenin were incubated in phosphate buffer saline. Protein quantification and morphological and mechanical analyses were conducted on the degraded scaffolds, and the incubated phosphate buffer saline was also tested for protein content. Fresh scaffolds were incubated overnight in conventional simulated body fluid to evaluate mineralization potential of the incorporated electrospun amelogenin. Human dermal fibroblasts were seeded onto scaffolds, incubated overnight, cryosectioned, and stained with 4′,6-diamidino-2-phenylindole to determine cellular adhesive properties. The incorporation of 5 mg/mL amelogenin into electrospun scaffolds improved mechanical properties (in poly(ϵ-caprolactone) scaffolds), increased fiber mineralization (in poly(glycolic acid) scaffolds), and improved human dermal fibroblast adhesion (in poly(ϵ-caprolactone) scaffolds). The presented results suggest that amelogenin can be used for multiple tissue engineering applications in the form of an additive to an electrospun scaffold.