Nanostructured hydroxyapatite surfaces-mediated adsorption alters recognition of BMP receptor IA and bioactivity of bone morphogenetic protein-2

Highly efficient loading of bone morphogenetic protein-2 (BMP-2) onto carriers with desirable performance is still a major challenge in the field of bone regeneration. Till now, the nanoscaled surface-induced changes of the structure and bioactivity of BMP-2 remains poorly understood. Here, the effect of nanoscaled surface on the adsorption and bioactivity of BMP-2 was investigated with a series of hydroxyapatite surfaces (HAPs): HAP crystal-coated surface (HAP), HAP crystal-coated polished surface (HAP-Pol), and sintered HAP crystal-coated surface (HAP-Sin). The adsorption dynamics of recombinant human BMP-2 (rhBMP-2) and the accessibility of the binding epitopes of adsorbed rhBMP-2 for BMP receptors (BMPRs) were examined by a quartz crystal microbalance with dissipation. Moreover, the bioactivity of adsorbed rhBMP-2 and the BMP-induced Smad signaling were investigated with C2C12 model cells. A noticeably high mass-uptake of rhBMP-2 and enhanced recognition of BMPR-IA to adsorbed rhBMP-2 were found on the HAP-Pol surface. For the rhBMP-2-adsorbed HAPs, both ALP activity and Smad signaling increased in the order of HAP-Sin < HAP < HAP-Pol. Furthermore, hybrid molecular dynamics and steered molecular dynamics simulations validated that BMP-2 tightly anchored on the HAP-Pol surface with a relative loosened conformation, but the HAP-Sin surface induced a compact conformation of BMP-2. In conclusion, the nanostructured HAPs can modulate the way of adsorption of rhBMP-2, and thus the recognition of BMPR-IA and the bioactivity of rhBMP-2. These findings can provide insightful suggestions for the future design and fabrication of rhBMP-2-based scaffolds/implants.Statement of SignificanceThis study provides strong evidences that nanoscaled HAPs yield extraordinary influence on the adsorption behaviors and bioactivity of rhBMP-2. It has been found that the surface roughness and crystallinity played a crucial role in governing the way of rhBMP-2 binding to HAPs, and thus the conformation, recognition of BMPR-IA and bioactivity of adsorbed rhBMP-2. It is also for the first time to correlate numerical modeling and experimental results of the bioactivity of rhBMP-2 on nanostructured HAPs. This work can pave an avenue for the wider uses of rhBMP-2 in clinical applications and arouse broad interests among researchers in the fields of nano-biotechnology, biomaterials and bone tissue engineering.

Nanostructured HAPs induced different adsorption states (adsorption amount and way of binding) of rhBMP-2, and thus the BMPRs-binding availability and bioactivity of adsorbed rhBMP-2. The rough surface (HAP-Pol) not only showed the highest mass-uptake of rhBMP-2, but also achieved an enhanced recruitment of BMPR-IA and up-regulated bioactivity of rhBMP-2.Figure optionsDownload high-quality image (128 K)Download as PowerPoint slide