Development and evaluation of bioactive 3D zein and zein/nano-hydroxyapatite scaffolds for bone tissue engineering application

The aim of this study is to generate and investigate biodegradable and biocompatible zein and zein/nano-hydroxyapatite composite scaffolds for bone defect healing. 3D zein scaffold was successfully fabricated using the salt-leaching method and incorporated with 12.5 wt% nHA for osteogenic differentiation of murine myoblast cell line (C2C12 cells). The scaffolds were subjected to physicochemical and biomechanical characterizations using the scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), biodegradation, porosity, mechanical tests. C2C12 cells were cultured on scaffolds and incubated for 21 days. Cell proliferation was detected by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Quantitative real-time PCR was used to test the expression of osteoblastic-related genes including Runx2, ALP, and Col1A1. The scaffolds had an adequate mean pore size and a total porosity of 61.1%–70.6%. The addition of 12.5 wt% nHA to the zein scaffold increased the compressive modulus to 79.1 MPa and the ultimate strength to 2.7 MPa. The qRT-PCR analysis confirmed that mRNA transcript levels were significantly higher ( p < 0.05) on the zein/nHA than on the pure zein scaffold. The results suggested that the developed scaffolds could be a potential candidate for bone tissue engineering due to their promising osteoinductivity, surface topography, mechanical behavior, biodegradability.