Extracellular matrix deposition and scaffold biodegradation in an in vitro three-dimensional model of bone by X-ray computed microtomography

Abstract

The development of an in vitro model of bone and the optimization of tools for determining the biological processes occurring during bone repair remains a major goal in the field of bone tissue engineering. Recently, a model based on a three-dimensional co-culture of osteoblasts and osteoclast precursors in Skelite(TM) scaffolds was developed. Although induction of osteoblast and osteoclast differentiation was observed, a complete evaluation of bone deposition and biodegradation processes was missing due to technical limitations. In the current study, both X-ray computed microtomography and histological analysis were used to monitor these two key biological processes in the same in vitro model. Either osteoblasts or a combination of osteoblasts and osteoclasts were seeded on Skelite(TM) scaffolds. Scaffold biodegradation and increased bone deposition together with a more organized extracellular matrix were observed in the co-cultures, highlighting the role of osteoclasts in the determination and regulation of bone deposition. Results confirmed the potential and relevance of co-culturing osteoblasts and osteoclasts to resemble native tissue. The combination of X-ray computed microtomography and histology presented in this study could be useful in future studies for the validation and development of new in vitro culture systems for bone tissue engineering.

Keywords: X-ray computed microtomography; bone remodelling; bone tissue engineering; histomorphometry; osteoblast; osteoclast; scaffold.