Gene profiling of mouse osteoblasts on demineralized bone paper

Osteoblasts play a crucial role in orchestrating osteoclast function during the bone remodeling cycle. However, current osteo culture and assay platforms face limitations in faithfully reproducing the intricate processes of bone remodeling. To address this challenge, we engineered demineralized bone paper (DBP), drawing inspiration from osteoid, to faithfully recapitulate the intrinsic phenotypes of osteoblasts. DBP demonstrated a remarkable capacity to support rapid and structurally significant mineral deposition by osteoblasts. Moreover, under the influence of vitamin D3 (VD3) and prostaglandin E2 (PGE2), osteoblasts cultured on DBP successfully mimicked in vivo-relevant osteoclastogenesis. To gain deeper insights into the molecular signatures underlying these phenotypic changes, we conducted RNA sequencing to characterize the gene expression profile of osteoblasts on DBP. The overall study design involved a comparative gene expression profiling analysis of osteoblasts cultured on a traditional tissue culture plate and those on DBP, with and without the influence of VD3 and PGE2. This comprehensive approach aimed to enhance our understanding of how DBP can serve as a more physiologically relevant platform for investigating the intricate interplay between osteoblasts and osteoclasts in bone remodeling processes.