Biological Characteristics of Mesenchymal Stem Cells Grown on Different Topographical Nanofibrous Poly-L-Lactide Meshes

Abstract

The nanotopographical features of artificial scaffolds have complex effects on the biological characteristics of stem cells. They influence cell adhesion, spreading, proliferation, and differentiation; however we have limited knowledge on how these processes occur under nanotopographical cues. In this study, two kinds of electrospun nanofibrous meshes with different fiber arrangements (totally non-woven and lattice-like) were fabricated and used for in vitro culture of mesenchymal stem cells (MSCs). By comparing the characteristic marks related to osteogenic differentiation, we found that with prolonged culture time, osteopontin (OPN), osteocalcin (OCN) and alkaline phosphatase (ALP), as well as related genes (Runx2 and Colla genes), were all expressed at higher levels on lattice-like nanofibrous meshes than on non-woven ones. These results indicated that the lattice-like nanofibrous mesh activated the osteogenic differentiation of MSCs owing to changes in cell morphology directed by nanofiber orientations. Compared with pure non-woven nanofibrous meshes, lattice-like ones possessed a combined structure of parallel, magnetic-line-like, and non-woven regions. MSCs adhering onto them had upregulated expression levels of integrin subunits a5 and b1, and activated downstream signaling pathways of Ras homolog gene family member A (RhoA) and extracellular signal-regulated kinase (ERK). When the specific inhibitors PD98059 and Y27632 were used to inhibit phosphorylated ERK and p160 ROCKII activity, respectively, F-actin became disordered and the expression level of Runx2 was downregulated. Thus, we concluded that the scaffold nanotopography may modulate the microenvironment of MSCs and promote their osteogenic differentiation through the RhoA and ERK signaling pathways. These findings provided valuable information on the selection of artificial matrices suitable for MSCs application in bone tissue engineering.