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.

Lack of osteointegration is a major cause of aseptic loosening and failure of implants used in bone replacement. Implants coated with angiogenic biomaterials can improve osteointegration and potentially reduce these complications. Silicon-and phosphorus-based materials have been shown to upregulate expression of angiogenic factors and improve endothelial cell functions. In the present study, we hypothesize that implants coated with amorphous silica-based coatings in the form of silicon oxynitrophosphide (SiONP) by using plasmaenhanced chemical vapor deposition (PECVD) technique could enhance human umbilical vein endothelial cell angiogenic properties in vitro. The tested groups were: glass coverslip (GCS), tissue culture plate, SiON, SiONP1 (O: 7.3 at %), and SiONP2 (O: 14.2 at %) implants. The SiONP2 composition demonstrated 3.5-fold more fibronectin deposition than the GCS ( p < 0.001). The SiONP2 group also presented a significant improvement in the capillary tubule length and thickness compared with the other groups ( p < 0.01). At 24 h, we observed at least a twofold upregulation of vascular endothelial growth factor A, hypoxia-inducible factor-1a, angiopoietin-1, and nesprin-2, more evident in the SiONP1 and SiONP2 groups. In conclusion, the studied amorphous silica-coated implants, especially the SiONP2 composition, could enhance the endothelial cell angiogenic properties in vitro and may induce faster osteointegration and healing.

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Volumetric muscle loss injuries overwhelm the endogenous regenerative capacity of skeletal muscle, and the associated oxidative damage can delay regeneration and prolong recovery. This study aimed to investigate the effect of silicon-ions on C2C12 skeletal muscle cells under normal and excessive oxidative stress conditions to gain insights into its role on myogenesis during the early stages of muscle regeneration. In vitro studies indicated that 0.1 mM Si-ions into cell culture media significantly increased cell viability, proliferation, migration, and myotube formation compared to control. Additionally, MyoG, MyoD, Neurturin, and GABA expression were significantly increased with addition of 0.1, 0.5, and 1.0 mM of Si-ion for 1 and 5 days of C2C12 myoblast differentiation. Furthermore, 0.1–2.0 mM Si-ions attenuated the toxic effects of H2O2 within 24 h resulting in increased cell viability and differentiation. Addition of 1.0 mM of Si-ions significantly aid cell recovery and protected from the toxic effect of 0.4 mM H2O2 on cell migration. These results suggest that ionic silicon may have a potential effect in unfavorable situations where reactive oxygen species is predominant affecting cell viability, proliferation, migration, and differentiation. Furthermore, this study provides a guide for designing Si-containing biomaterials with desirable Si-ion release for skeletal muscle regeneration.

Critical sized bone defects are challenging to heal due to the sudden and large volume of lost bone. Fixative plates are often used to stabilize defects, yet oxidative stress and delayed angiogenesis are contributing factors to poor biocompatibility and delayed bone healing. This study tests the angiogenic and antioxidant properties of amorphous silicon oxynitrophosphide (SiONPx) nanoscale coating material on endothelial cells to regenerate vascular tissue in vitro and in bone defects. In vitro studies evaluate the effect of SiONx and two different SiONPx compositions on human endothelial cells exposed to reactive oxygen species (e.g., H2O2) that simulates oxidative stress conditions. In vivo studies using adult male Sprague Dawley rats (~450 g) were performed to compare a bare plate, SiONPx-coated implant plate, and a sham control group using a rat standard sized calvarial defect. Results from this study showed that plates coated with SiONPx significantly reduced cell death, enhanced vascular tubule formation and matrix deposition by upregulating angiogenic and antioxidant expression (e.g. VEGFA, angiopoetin-1, SOD-1, NRF-2 and Cat-1). Moreover, endothelial cell markers (CD31) demonstrate a significant tubular structure on the SiONPx coating group compared to an empty and uncoated plate group. This reveals that atomic doping of phosphate into the nanoscale coating of SiONx produced markedly elevated levels of antioxidant and angiogenic markers that enhance vascular tissue regeneration. This study demonstrates that SiONPx or SiONx nanoscale coated materials enhance antioxidant expression, angiogenic marker expression, and reduce ROS levels needed for accelerating vascular tissue regeneration. These results further suggest that SiONPx nanoscale coating could be a promising candidate for titanium plate for rapid and enhanced cranial bone defect healing.

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