Silicon nitride enhances osteoprogenitor cell growth and differentiation via increased surface energy and formation of amide and nanocrystalline HA for craniofacial reconstruction

Abstract

The bioactive silicon nitride (Si3N4) has been FDA cleared for use as spinal intervertebral arthrodesis devices. Because its surface properties promote bone ongrowth and ingrowth, it also has the potential to benefit craniofacial reconstruction. Thus, the aim of this work was to determine whether the surface properties of Si3N4 could enhance the osteoblast cell growth, differentiation and nucleation of hydroxyapatite (HA) crystals compared to conventional implant materials such as titanium (Ti) and polyether ether ketone (PEEK). X-ray absorbance near-edge structure analysis (XANES) indicated the presence of Si-Si, Si-O and Si-N bonding. Surface wettability studies confirmed that Si3N4 exhibits the lowest contact angle and highest surface energy. Cell culture studies showed that osteoblast growth was enhanced on Si3N4 after 1 day and up to 7 days. Si3N4 surface induced highest surface coverage and thickness of nanocrystalline HA (211) and (203) in cell-free in vitro studies after 7 days of culture. Raman spectroscopy analysis confirmed the presence of surface functional groups consisting of phosphate and carbonate species. Interestingly, Si3N4 surface showed amide and hydroxyproline groups, the precursors to collagen, which were not observed on Ti and PEEK surfaces. Furthermore, Si3N4 surface indicated high expression of RUNX2, enhanced cell differentiation and dense collagenous ECM after 30 days of the in vitro study. The present study concluded that Si3N4 surface enhances osteoprogenitor cell adhesion, growth, RUNX2 expression and ECM formation via the coupled effects of higher surface energy and the presence of amide and nanocrystalline HA functional groups.