rhBMP-2 Induces Immature Muscular Tissue to Differentiate into Bone-Like Tissue In Vitro

Abstract

The aim of this study is to induce bone from immature muscular tissue in vitro using recombinant human BMP (rhBMP)-2 and expanded polytetrafluoroethylene (ePTFE) as a scaffold. Commercially available rhBMP-2 was used in this experiment. IMTs were harvested from the forelimbs of 20th Sprague-Dawley embryonic rats and placed into a homogenizer with 10ng/μl of rhBMP-2 and then homogenized. The homogenized IMT was placed on ePTFE and cultured for 2 weeks. The analyses of histological observation, electron probe micro analyzer (EPMA), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) were carried out following culture. The bone-like tissue, which was made up of osteoblast-like cells and osteoids, was partially observed by H-E staining. Moreover, strong mineral deposition was observed in the extracellular matrix by von Kossa staining. Ca, P and O were detected in the extracellular matrix by EPMA and were confirmed to be at almost the same position based on the findings of synchronized images. XRD patterns and FTIR spectra of specimen were found to have typical hydroxyapatite crystal peaks and spectra, respectively. These results suggest that rhBMP-2 induced IMT differentiation into bone-like tissue in vitro.