SiONx Coating Regulates Mesenchymal Stem Cell Antioxidant Capacity via Nuclear Erythroid Factor 2 Activity under Toxic Oxidative Stress Conditions.

Healing in compromised and complicated bone defects is often prolonged and delayed due to the lack of bioactivity of the fixation device, secondary infections, and associated oxidative stress. Here, we propose amorphous silicon oxynitride (SiONx) as a coating for the fixation devices to improve both bioactivity and bacteriostatic activity and reduce oxidative stress. We aimed to study the effect of increasing the N/O ratio in the SiONx to fine-tune the cellular activity and the antioxidant effect via the NRF2 pathway under oxidative stress conditions. The in vitro studies involved using human mesenchymal stem cells (MSCs) to examine the effect of SiONx coatings on osteogenesis with and without toxic oxidative stress. Additionally, bacterial growth on SiONx surfaces was studied using methicillin-resistant Staphylococcus aureus (MRSA) colonies. NRF2 siRNA transfection was performed on the hMSCs (NRF2-KD) to study the antioxidant response to silicon ions. The SiONx implant surfaces showed a >4-fold decrease in bacterial growth vs. bare titanium as a control. Increasing the N/O ratio in the SiONx implants increased the alkaline phosphatase activity >1.5 times, and the other osteogenic markers (osteocalcin, RUNX2, and Osterix) were increased >2-fold under normal conditions. Increasing the N/O ratio in SiONx enhanced the protective effects and improved cell viability against toxic oxidative stress conditions. There was a significant increase in osteocalcin activity compared to the uncoated group, along with increased antioxidant activity under oxidative stress conditions. In NRF2-KD cells, there was a stunted effect on the upregulation of antioxidant markers by silicon ions, indicating a role for NRF2. In conclusion, the SiONx coatings studied here displayed bacteriostatic properties. These materials promoted osteogenic markers under toxic oxidative stress conditions while also enhancing antioxidant NRF2 activity. These results indicate the potential of SiONx coatings to induce in vivo bone regeneration in a challenging oxidative stress environment.

Revolutionizing bone regeneration: advanced biomaterials for healing compromised bone defects.

In this review, we explore the application of novel biomaterial-based therapies specifically targeted towards craniofacial bone defects. The repair and regeneration of critical sized bone defects in the craniofacial region requires the use of bioactive materials to stabilize and expedite the healing process. However, the existing clinical approaches face challenges in effectively treating complex craniofacial bone defects, including issues such as oxidative stress, inflammation, and soft tissue loss. Given that a significant portion of individuals affected by traumatic bone defects in the craniofacial area belong to the aging population, there is an urgent need for innovative biomaterials to address the declining rate of new bone formation associated with age-related changes in the skeletal system. This article emphasizes the importance of semiconductor industry-derived materials as a potential solution to combat oxidative stress and address the challenges associated with aging bone. Furthermore, we discuss various material and autologous treatment approaches, as well as in vitro and in vivo models used to investigate new therapeutic strategies in the context of craniofacial bone repair. By focusing on these aspects, we aim to shed light on the potential of advanced biomaterials to overcome the limitations of current treatments and pave the way for more effective and efficient therapeutic interventions for craniofacial bone defects.

Mini review: Biomaterials in repair and regeneration of nerve in a volumetric muscle loss.

Volumetric muscle loss (VML) following a severe trauma or injury is beyond the intrinsic regenerative capacity of muscle tissues, and hence interventional therapy is required. Extensive muscle loss concomitant with damage to neuromuscular components overwhelms the muscles' remarkable regenerative capacity. The loss of nervous and vascular tissue leads to further damage and atrophy, so a combined treatment for neuromuscular junction (NMJ) along with the volumetric muscle regeneration is important. There have been immense advances in the field of tissue engineering for skeletal muscle tissue and peripheral nerve regeneration, but very few address the interdependence of the tissues and the need for combined therapies to repair and regenerate fully functional muscle tissue. This review addresses the problem and presents an overview of the biomaterials that have been studied for tissue engineering of neuromuscular tissues associated with skeletal muscles.

Silicon Oxynitrophosphide Nanoscale Coating Enhances Antioxidant Marker-Induced Angiogenesis During in vivo Cranial Bone-Defect Healing.

Critical-sized bone defects are challenging to heal because of 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 silicon oxynitride (SiONx) and two different SiONPx compositions on human endothelial cells exposed to ROS (eg, hydrogen peroxide) that simulates oxidative stress conditions. in vivo studies using adult male Sprague Dawley rats (approximately 450 g) were performed to compare a bare plate, a 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, and enhanced vascular tubule formation and matrix deposition by upregulating angiogenic and antioxidant expression (eg, vascular endothelial growth factor A, angiopoetin-1, superoxide dismutase 1, nuclear factor erythroid 2-related factor 2, and catalase 1). Moreover, endothelial cell markers (CD31) showed a significant tubular structure in the SiONPx coating group compared with 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 found 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. © 2020 The Authors. JBMR Plus published by Wiley Periodicals LLC. on behalf of American Society for Bone and Mineral Research.

Bioprinted nanocomposite hydrogels: A proposed approach to functional restoration of skeletal muscle and vascular tissue following volumetric muscle loss.

Musculoskeletal conditions are the highest contributor to global disability, accounting for 16% of all ages lived with disability. Volumetric muscle loss (VML) is classified as significant damage to skeletal muscle compartments and motor units, leading to significant tissue loss, functional deficits, and long-term disability. In this review, the current tissue engineering approaches in terms of fabrication techniques, materials, cell sources, and growth factors for enhanced angiogenesis and neuromuscular junction (NMJ) in VML repair, are discussed. Review of results recently published in the literature suggested that bioprinted nanocomposite hydrogels (NC gels) seeded with adult muscle progenitor cells that promote secretion of endogenous vascular growth factors have potential applications in promoting skeletal muscle regeneration, revascularization, and NMJ repair (Figure 1). Despite recent advancements, future research is needed on NC gels and the complex processes underlying vascular infiltration and NMJ repair in VML injuries.

Ionic Silicon Protects Oxidative Damage and Promotes Skeletal Muscle Cell Regeneration.

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.

A comparative study on silicon nitride, titanium and polyether ether ketone on mouse pre-osteoblast cells.

The current study provides more insights about the surface bioactivity of the silicon nitride (Si3N4) as a potential candidate for bone regeneration in craniofacial and orthopaedic applications compared with conventional implantation materials. Current skeletal reconstructive materials such as titanium and polyether ether ketone (PEEK) are limited by poor long-term stability, biocompatibility and prolonged healing. Si3N4 is an FDA-approved material for an intervertebral spacer in spinal fusion applications. It is biocompatible and has antimicrobial properties. Here, we hypothesize that Si3N4 was found to be an osteoconductive material and conducts the growth, differentiation of MC3T3-E1 cells for extracellular matrix deposition, mineralization and eventual bone regeneration for craniofacial and orthopaedic applications. MC3T3-E1 cells were used to study the osteoblastic differentiation and mineralization on sterile samples of Si3N4, titanium alloy and PEEK. The samples were then analysed for extracellular matrix deposition and mineralization by FTIR, Raman spectroscopy, SEM, EDX, Alizarin Red, qRT-PCR and ELISA. The in vitro study indicates the formation of collagen fibres and mineral deposition on all three sample surfaces. There was more profound and faster ECM deposition and mineralization on Si3N4 surface as compared to titanium and PEEK. The FTIR and Raman spectroscopy show formation of collagen and mineral deposition at 30 days for Si3N4 and titanium and not PEEK. The peaks shown by Raman for Si3N4 resemble closely to natural bone. Results also indicate the upregulation of osteogenic transcription factors such as RUNX2, SP7, collagen type I and osteocalcin. The authors concluded that Si3N4 rapidly conducts mineralized tissue formation via extracellular matrix deposition and biomarker expression in mouse calvarial pre-osteoblast cells. Thus, this study confirms that the bioactive Si3N4 could be a potential material for craniofacial and orthopaedic applications leading to rapid bone regeneration that resemble the natural bone structure.

Novel 3D-printed methacrylated chitosan-laponite nanosilicate composite scaffolds enhance cell growth and biomineral formation in MC3T3 pre-osteoblasts.

This study compared the effect of gelatin- and chitosan-based scaffolds on osteoblast biomineralization. These scaffolds have been modified using methacrylate and laponite nanosilicates to improve their mechanical strength and support osteoblast function. Scaffold materials were prepared to have the same compressive strength (14-15 MPa) such that differences in cell response would be isolated to differences in biopolymer chemistry. The materials were tested for rheological properties to optimize the bio-ink for successful 3D printing using a robocast-assisted deposition system. Osteoblasts were cultured on the surface of 3D-printed methacrylated chitosan-laponite (MAC-Lp), methacrylated gelatin-laponite (MAG-Lp), MAC, and MAG scaffolds. MAC-Lp scaffolds showed increased cell viability, cell growth, and biomineral formation as compared to MAG-Lp scaffolds. FTIR results showed the presence of higher biomineral phosphate and extracellular matrix (ECM) collagen-like amide formation on MAC-Lp scaffolds as compared to MAG-Lp scaffolds. MAC-Lp scaffolds showed increased density of ECM-like tissue from SEM analysis, stained mineral nodules from Alizarin staining, and the existence of Ca─P species evident by X-ray absorbance near edge structure analysis. In conclusion, MAC-Lp scaffolds enhanced osteoblast growth and biomineral formation as compared to MAG-Lp scaffolds.

Amorphous Silicon Oxynitrophosphide-Coated Implants Boost Angiogenic Activity of Endothelial Cells.

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 plasma-enhanced 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-1α, 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. Impact Statement In this study, we report for the first time the significant enhancement of human umbilical vein endothelial cell angiogenic properties (in vitro) by the amorphous silica-based coatings in the form of silicon oxynitrophosphide (SiONP). The SiONP2 demonstrated 3.5-fold more fibronectin deposition than the glass coverslip and presented a significant improvement in the capillary tubule length and thickness. At 24 h, SiONP reported twofold upregulation of vascular endothelial growth factor A, hypoxia-inducible factor-1α, angiopoietin-1, and nesprin-2. The studied amorphous silica-coated implants enhance the endothelial cell angiogenic properties in vitro and may induce faster osteointegration and healing.

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

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.

Injectable scaffolds: Preparation and application in dental and craniofacial regeneration.

Injectable scaffolds are appealing for tissue regeneration because they offer many advantages over pre-formed scaffolds. This article provides a comprehensive review of the injectable scaffolds currently being investigated for dental and craniofacial tissue regeneration. First, we provide an overview of injectable scaffolding materials, including natural, synthetic, and composite biomaterials. Next, we discuss a variety of characteristic parameters and gelation mechanisms of the injectable scaffolds. The advanced injectable scaffolding systems developed in recent years are then illustrated. Furthermore, we summarize the applications of the injectable scaffolds for the regeneration of dental and craniofacial tissues that include pulp, dentin, periodontal ligament, temporomandibular joint, and alveolar bone. Finally, our perspectives on the injectable scaffolds for dental and craniofacial tissue regeneration are offered as signposts for the future advancement of this field.