Objective: The aim of this study is to induce bone from immature muscular tissue in vitrousing recombinant human BMP(rhBMP)-2, rhBMP-4 and rhBMP-7.
Method: Commercially available rhBMP-2, rhBMP-4 and rhBMP-7 were used in this experiment. IMTs were harvested from the forelimbs of 20th Sprague-Dawley embryonic rats and placed into a homogenizer with 10ng/μl each of rhBMP-2, rhBMP-4 and rhBMP-7 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(FT-IR) were carried out following culture.
Result: 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 FT-IR spectra of specimen were found to have typical apatite crystal peaks and spectra, respectively.
Conclusion: These results suggest that rhBMP-2, rhBMP-4 and rhBMP-7 induced immature muscular tissue differentiation into bone-like tissue in vitro, and thus, immature muscular tissue has a possibility to be an effective tissue source for bone tissue engineering.