Abstract
The purpose of the study is to investigate the effects of electrospun fiber diameter and orientation on differentiation and ECM organization of bone marrow stromal cells (BMSCs), in attempt to provide rationale for fabrication of a periosteum mimetic for bone defect repair. Cellular growth, differentiation, and ECM organization were analyzed on PLGA-based random and aligned fibers using fluorescent microscopy, gene analyses, electron scanning microscopy (SEM), and multiphoton laser scanning microscopy (MPLSM). BMSCs on aligned fibers had a reduced number of ALP+ colony at Day 10 as compared to the random fibers of the same size. However, the ALP+ area in the aligned fibers increased to a similar level as the random fibers at Day 21 following stimulation with osteogenic media. Compared with the random fibers, BMSCs on the aligned fibers showed a higher expression of OSX and RUNX2. Analyses of ECM on decellularized spun fibers showed highly organized ECM arranged according to the orientation of the spun fibers, with a broad size distribution of collagen fibers in a range of 40-2.4 μm. Taken together, our data support the use of submicron-sized electrospun fibers for engineering of oriented fibrous tissue mimetic, such as periosteum, for guided bone repair and reconstruction.