Enhancing osteoblast differentiation through small molecule-incorporated engineered nanofibrous scaffold

Objective: This study aimed to investigate the effect of small molecules incorporated into the engineered nanofibrous scaffold to enhance the osteoblast differentiation Materials and methods: Poly-ε-caprolactone (PCL) nanofiber matrices with lithium chloride (LiCl) were fabricated using the electrospinning technique. Scaffolds were characterized using scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX). Scaffolds were seeded with MC3T3-E1 cells and assessed using Western blots (β-catenin), alamarBlue assay (proliferation), qPCR (osteoblast differentiation), and mineralization (Alizarin Red staining). Results: We observed LiCl nanofiber scaffolds induced concentration-dependent cell proliferation that correlated with an increased β-catenin expression indicating sustained Wnt signaling. Next, we examined osteoblast differentiation markers such as osteocalcin (OCN) and Runt-related transcription factor 2 (Runx2) and noted increased expression in LiCl nanofiber scaffolds. We also noted increased bone morphogenetic protein (BMP-2, 4, and 7) expressions suggesting activated Wnt can promote cures to further osteogenic differentiation. Finally, Alizarin Red staining demonstrated increased mineral deposition in LiCl-incorporated nanofiber scaffolds. Conclusions: Together, these results indicated that LiCl-incorporated nanofiber scaffolds enhance osteoblast differentiation. Clinical relevance: Small molecule-incorporated nanofibrous scaffolds are an innovative clinical tool for bone tissue engineering.