β-TCP Scaffolds Research Articles

Metal-Organic Framework Functionalized Bioceramic Scaffolds with Antioxidative Activity for Enhanced Osteochondral Regeneration.

Osteoarthritis (OA) is a degenerative disease that often causes cartilage lesions and even osteochondral damage. Osteochondral defects induced by OA are accompanied by an inflammatory arthrosis microenvironment with overproduced reactive oxygen species (ROS), resulting in the exacerbation of defects and difficulty regenerating osteochondral tissues. Therefore, it is urgently needed to develop osteochondral scaffolds that can not only promote the integrated regeneration of cartilage and subchondral bone, but also possess ROS-scavenging ability to protect tissues from oxidative stress. Herein, zinc-cobalt bimetallic organic framework (Zn/Co-MOF) functionalized bioceramic scaffolds are designed for repairing osteochondral defects under OA environment. By functionalizing Zn/Co-MOF on the 3D-printed beta-tricalcium phosphate (β-TCP) scaffolds, the Zn/Co-MOF functionalized β-TCP (MOF-TCP) scaffolds with broad-spectrum ROS-scavenging ability are successfully developed. Benefiting from its catalytic active sites and degradation products, Zn/Co-MOF endows the scaffolds with excellent antioxidative and anti-inflammatory properties to protect cells from ROS invasion, as well as dual-bioactivities of simultaneously inducing osteogenic and chondrogenic differentiation in vitro. Furthermore, in vivo results confirm that MOF-TCP scaffolds accelerate the integrated regeneration of cartilage and subchondral bone in severe osteochondral defects. This study offers a promising strategy for treating defects induced by OA as well as other inflammatory diseases.

3D printed mesoporous bioactive glass, bioglass 45S5, and β-TCP scaffolds for regenerative medicine: A comparative invitro study.

While autografts to date remain the "gold standard" for bone void fillers, synthetic bone grafts have garnered attention due to their favorable advantages such as ability to be tailored in terms of their physical and chemical properties. Bioactive glass (BG), an inorganic material, has the capacity to form a strong bond with bone by forming a bone-like apatite surface, enhancing osteogenesis. Coupled with additive manufacturing (3D printing) it is possible to maximize bone regenerative properties of the BG. The objective of this study was to synthesize and characterize 3D printed mesoporous bioactive glass (MBG), BG 45S5, and compare to β-Tricalcium phosphate (β-TCP) based scaffolds; test cell viability and osteogenic differentiation on human osteoprogenitor cells invitro. MBG, BG 45S5, and β-TCP were fabricated into colloidal gel suspensions, tested with a rheometer, and manufactured into scaffolds using a 3D direct-write micro-printer. The materials were characterized in terms of microstructure and composition with Thermogravimetric Analyzer/Differential Scanning Calorimeter (TGA/DSC), Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Micro-Computed Tomography (μ-CT), Scanning Electron Microscopy (SEM), Energy Dispersive X-ray Spectroscopy (EDS), and Mattauch-Herzog-Inductively Coupled Plasma-Mass Spectrometry (MH-ICP-MS). Scaffolds were tested for cell proliferation and osteogenic differentiation using human osteoprogenitor cells. Osteogenic media was used for differentiation, and immunocytochemistry for osteogenic markers Runx-2, Collagen-I, and Osteocalcin. The cell viability results after 7 days of culture yielded significantly higher (p < 0.05) results in β-TCP scaffolds compared to BG 45S5 and MBG groups. All materials expressed osteogenic markers after 21 days of culture in expansion and osteogenic media.

Customized bioceramic scaffolds and metal meshes for challenging large-size mandibular bone defect regeneration and repair.

Large-size mandible graft has huge needs in clinic caused by infection, tumor, congenital deformity, bone trauma and so on. However, the reconstruction of large-size mandible defect is challenged due to its complex anatomical structure and large-range bone injury. The design and fabrication of porous implants with large segments and specific shapes matching the native mandible remain a considerable challenge. Herein, the 6% Mg-doped calcium silicate (CSi-Mg6) and β- and α-tricalcium phosphate (β-TCP, α-TCP) bioceramics were fabricated by digital light processing as the porous scaffolds of over 50% in porosity, while the titanium mesh was fabricated by selective laser melting. The mechanical tests showed that the initial flexible/compressive resistance of CSi-Mg6 scaffolds was markedly higher than that of β-TCP and α-TCP scaffolds. Cell experiments showed that these materials all had good biocompatibility, while CSi-Mg6 significantly promoted cell proliferation. In the rabbit critically sized mandible bone defects (∼13 mm in length) filled with porous bioceramic scaffolds, the titanium meshes and titanium nails were acted as fixation and load bearing. The results showed that the defects were kept during the observation period in the blank (control) group; in contrast, the osteogenic capability was significantly enhanced in the CSi-Mg6 and α-TCP groups in comparison with the β-TCP group, and these two groups not only had significantly increased new bone formation but also had thicker trabecular and smaller trabecular spacing. Besides, the CSi-Mg6 and α-TCP groups showed appreciable material biodegradation in the later stage (from 8 to 12 weeks) in comparison with the β-TCP scaffolds while the CSi-Mg6 group showed much outstanding mechanical capacity in vivo in the early stage compared to the β-TCP and α-TCP groups. Totally, these findings suggest that the combination of customized strength-strong bioactive CSi-Mg6 scaffolds together with titanium meshes is a promising way for repairing the large-size load-bearing mandible defects.

Polydopamine-coated 3D-printed β-tricalcium phosphate scaffolds to promote the adhesion and osteogenesis of BMSCs for bone-defect repair: mRNA transcriptomic sequencing analysis.

Cellular bioactivity and tissue regeneration can be affected by coatings on tissue-engineered scaffolds. Using mussel-inspired polydopamine (PDA) is a convenient and effective approach to surface modification. Therefore, 3D-printed β-tricalcium phosphate (β-TCP) scaffolds were coated with PDA in this study. The effects of the scaffolds on the adhesion and osteogenic differentiation of seeded bone marrow mesenchymal stem cells (BMSCs) in vitro and on new-bone formation in vivo were investigated. The potential mechanisms and related differential genes were assessed using mRNA sequencing. It was seen that PDA coating increased the surface roughness of the 3D-printed β-TCP scaffolds. Furthermore, it prompted the adhesion and osteogenic differentiation of seeded BMSCs. mRNA sequencing analysis revealed that PDA coating might affect the osteogenic differentiation of BMSCs through the calcium signaling pathway, Wnt signaling pathway, TGF-beta signaling pathway, etc. Moreover, the expression of osteogenesis-related genes, such as R-spondin 1 and chemokine c-c-motif ligand 2, was increased. Finally, both the 3D-printed β-TCP scaffolds and PDA-coated scaffolds could significantly accelerate the formation of new bone in critical-size calvarial defects in rats compared with the control group; and the new bone formation was obviously higher in the PDA-coated scaffolds than in β-TCP scaffolds. In summary, 3D-printed β-TCP scaffolds with a PDA coating can improve the physicochemical characteristics and cellular bioactivity of the scaffold surface for bone regeneration. Potential differential genes were identified, which can be used as a foundation for further research.

Synergistic Effect of Incorporation of BG 45S5 and Silver Nanoparticles on β-TCP Scaffolds: Structural Characterization and Evaluation of Antimicrobial Activity and Biocompatibility

Synergistic Effect of Incorporation of BG 45S5 and Silver Nanoparticles on β-TCP Scaffolds: Structural Characterization and Evaluation of Antimicrobial Activity and Biocompatibility

Performance of Polydioxanone-Based Membrane in Association with 3D-Printed Bioceramic Scaffolds in Bone Regeneration

This study evaluated the bioactivity of 3D-printed β-tricalcium phosphate (β-TCP) scaffolds or hydroxyapatite (HA) scaffolds associated with polydioxanone (PDO) membrane (Plenum® Guide) for guided bone regeneration in rats. Fifty-four rats were divided into three groups (n = 18 animals): autogenous bone + PDO membrane (Auto/PG); 3D-printed β-TCP + PDO membrane (TCP/PG); and 3D-printed HA + PDO membrane (HA/PG). A surgical defect in the parietal bone was made and filled with the respective scaffolds and PDO membrane. The animals were euthanized 7, 30, and 60 days after the surgical procedure for micro-CT, histomorphometric, and immunolabeling analyses. Micro-CT showed an increase in trabecular thickness and a decrease in trabecular separation, even with similar bone volume percentages between TCP/PG and HA/PG vs. Auto/PG. Histometric analysis showed increased bone formation at 30 days in the groups compared to 7 days postoperatively. Immunolabeling analysis showed an increase in proteins related to bone formation at 30 days, and both groups showed a similar immunolabeling pattern. This study concludes that 3D-printed scaffolds associated with PDO membrane (Plenum® Guide) present similar results to autogenous bone for bone regeneration.

Efficacy of soluble lansoprazole-impregnated beta-tricalcium phosphate for bone regeneration

The proton pump inhibitor lansoprazole has been previously identified to upregulate the expression and transcriptional activity of runt-related transcription factor 2 (Runx2) that promotes lineage commitment and differentiation of osteoprogenitor cells. We could not elicit the expected efficacy of insoluble lansoprazole in enhancing osteogenesis when combined with beta-tricalcium phosphate (β-TCP) bone substitutes. This study aimed to evaluate the effects of soluble lansoprazole on in vitro osteoblastogenesis and new bone formation in vivo. Commercially available human mesenchymal stem cells or patient-derived bone marrow-derived stromal cells were treated with 20 µM of soluble lansoprazole at the beginning of osteogenic induction. Soluble lansoprazole-impregnated β-TCP materials were embedded in the cortical bone defect model of rabbits. Rabbits were sacrificed four weeks postoperatively and undecalcified bone specimens were prepared for evaluation of intra-material new bone formation. Only a 1-day treatment with soluble lansoprazole facilitated osteoblastic differentiation and matrix calcium deposition when added to undifferentiated human mesenchymal stromal cells at the beginning of the osteogenic differentiation. Soluble lansoprazole dose-dependently accelerated intra-material new bone formation when being impregnated with porous β-TCP artificial bones. Local use of soluble lansoprazole can be applicable for fracture and bone defect repair when combined with porous β-TCP scaffolds.

In vivo prevascularization strategy enhances neovascularization of β-tricalcium phosphate scaffolds in bone regeneration

BackgroundNeovascularization is critical for bone regeneration. Numerous studies have explored prevascularization preimplant strategies, ranging from calcium phosphate cement (CPC) scaffolds to co-culturing CPCs with stem cells. The aim of the present study was to evaluate an alternative in vivo prevascularization approach, using preimplant-prepared macroporous beta-tricalcium phosphate (β-TCP) scaffolds and subsequent transplantation in bone defect model.MethodsThe morphology of β-TCPs was characterized by scanning electron microscopy. After 3 weeks of prevascularization within a muscle pouch at the lateral size of rat tibia, we transplanted prevascularized macroporous β-TCPs in segmental tibia defects, using blank β-TCPs as a control. Extent of neovascularization was determined by angiography and immunohistochemical (IHC) evaluations. Tibia samples were collected at different time points for biomechanical, radiological, and histological analyses. RT-PCR and western blotting were used to evaluate angio- and osteo-specific markers.ResultsWith macroporous β-TCPs, we documented more vascular and supporting tissue invasion in the macroporous β-TCPs with prior in vivo prevascularization. Radiography, biomechanical, IHC, and histological analyses revealed considerably more vascularity and bone consolidation in β-TCP scaffolds that had undergone the prevascularization step compared to the blank β-TCP scaffolds. Moreover, the prevascularization treatment remarkably upregulated mRNA and protein expression of BMP2 and vascular endothelial growth factor (VEGF) during bone regeneration.ConclusionThis novel in vivo prevascularization strategy successfully accelerated vascular formation to bone regeneration. Our findings indicate that prevascularized tissue-engineered bone grafts have promising potential in clinical applications.The translational potential of this articleThis study indicates a novel in vivo prevascularization strategy for growing vasculature on β-TCP scaffolds to be used for repair of large segmental bone defects, might serve as a promising tissue-engineered bone grafts in the future.

Bioinspired sandwich-like hybrid surface functionalized scaffold capable of regulating osteogenesis, angiogenesis, and osteoclastogenesis for robust bone regeneration.

Recently, strategies that focus on biofunctionalized implant surfaces to enhance bone defect healing through the synergistic regulation of osteogenesis, angiogenesis, and osteoclastogenesis have attracted increasing attention in the bone tissue engineering field. Studies have shown that the Wnt/β-catenin signaling pathway has an imperative effect of promoting osteogenesis and angiogenesis while reducing osteoclastogenesis. However, how to prepare biofunctionalized bone implants with balanced osteogenesis, angiogenesis, and osteoclastogenesis by activating the Wnt/β-catenin pathway has seldom been investigated. Herein, through a bioinspired dopamine chemistry and self-assembly method, BML-284 (BML), a potent and highly selective Wnt signaling activator, was loaded on a mussel-inspired polydopamine (PDA) layer that had been immobilized on the porous beta-tricalcium calcium phosphate (β-TCP) scaffold surface and subsequently modified by a biocompatible carboxymethyl chitosan hydrogel to form a sandwich-like hybrid surface. β-TCP provides a biomimetic three-dimensional porous microenvironment similar to that of natural cancellous bone, and the BML-loaded sandwich-like hybrid surface endows the scaffold with multifunctional properties for potential application in bone regeneration. The results show that the sustained release of BML from the sandwich-like hybrid surface significantly facilitates the adhesion, migration, proliferation, spreading, and osteogenic differentiation of MC3T3-E1 cells as well as the angiogenic activity of human umbilical vein endothelial cells. In addition to osteogenesis and angiogenesis, the hybrid surface also exerts critical roles in suppressing osteoclastic activity. Remarkably, in a critical-sized cranial defect model, the biofunctionalized β-TCP scaffold could potentially trigger a chain of biological events: stimulating the polarization of M2 macrophages, recruiting endogenous stem cells and endothelial cells at the injury site to enable a favorable microenvironment for greatly accelerating bone ingrowth and angiogenesis while compromising osteoclastogenesis, thereby promoting bone healing. Therefore, these surface-biofunctionalized β-TCP implants, which regulate the synergies of osteogenesis, angiogenesis, and anti-osteoclastogenesis, indicate strong potential for clinical application as advanced orthopedic implants.

Immediate to short-term inflammatory response to biomaterial implanted in calvarium of mice.

Scaffolds made of biodegradable materials play a very important role in repairing bone defects. Our study was conducted with the aim of investigating inflammation, vascular changes, and tissue necrosis after the placement of 3D printed scaffolds composed of beta-tricalcium phosphate (TCP-β) on the calvarial bone defect of mice. Eight samples of scalp tissue in mice were examined in four groups (one-week control, two-week control, one-week experiment, and two-week experiment). Mice with routine bone defects were selected as the control group and mice with bone defects with β-TCP scaffolds were selected as the experimental group (TCP). The groups were evaluated in terms of inflammatory cells, osteoblast and osteoclast cells, vascular changes, and the number of resorption pit and empty lacuna. The results demonstrated a decrease in inflammatory cells and an increase in osteoclast and osteoblast cells in bone defect sites placed with TCP-β scaffolds (p<0.05). The results of histological staining showed pit resorption and further vascularization in the place of TCP-β scaffolds, but these changes were not statistically significant (p>0.05). Examining the number of empty lacunae in the bone defect site showed that TCP-β could significantly reduce the number of these lacunae in the bone defect sites placed with TCP-β scaffolds (p<0.05). 3D printed scaffolds composed of TCP-β that were implanted in bone defect sites were effective in reducing the inflammatory responses, emptying lacunae and increasing bone regeneration.

Magnesium dopped Calcium-Fluoride/Icaritin composite multi-layer coating functionalized 3D printed β-TCP scaffold induces sustained bone regeneration in a rabbit model

Osteonecrosis of femoral head(ONFH) requires bone grafts to fill bone defects and provide braced force after hip preservation surgery. Scaffolds functionalized with osteogenic factors may solve problems above. Icaritin (ICT) nanofilm and Magnesium dopped Calcium-Fluoride (Mg-CaF) nanofilm were loaded on β-TCP scaffolds by plasma coating technology and two new scaffolds were fabricated, namely ICT/β-TCP (IT) scaffold and Mg-CaF/ICT/β-TCP (MFIT) scaffold. In vitro,physical characterization, cytotoxicity, osteogenic ability and ICT release of scaffolds were assessed. Furthermore, osteogenic and angiogenic abilities of IT scaffolds and MFIT scaffolds, as well as their biocompatibility after implantation, were determined in ONFH rabbits. Results showed both scaffolds remained well-designed in structure and mechanical properties after coating. ICT release on MFIT scaffold was more sustained than on IT scaffold. Analysis of calcium nodules at 28 days indicated that degradation of Mg-CaF film promoted bone formation. Correspondingly, Micro-CT and histological staining suggested MFIT scaffold possessed similar osteogenic and angiogenic abilities compared with IT scaffold at 6 weeks and significantly better at 12,18 weeks. In addition, MFIT scaffold supported the weight-bearing area effectively throughout the observation period. Therefore, MFIT scaffolds provided stable support along with persistent osteogenesis and angiogenesis, which had potential applications inONFH.

A 3D-printed Sn-doped calcium phosphate scaffold for bone tissue engineering

Recent developments in 3D printing technology have been applied in the field of tissue engineering to fabricate customized bone repair scaffolds. β-tricalcium phosphate (β-TCP) is a bioceramic material with excellent potential as a scaffold foundation. Doping metallic ions with β-TCP will significantly enhance the mechanical property and bone regeneration performance compared with pure β-TCP specimens. In this study, we proposed a protocol for the fabrication of a Sn-doped β-TCP (Sn@TCP) scaffold using 3D printing technology, and the effect of Sn-doping on the physicochemical properties of the material and its in vitro bioactivity were investigated. Polyethylene glycol and polyvinyl alcohol were used as binder to construct Sn@TCP scaffolds which have good biocompability and can break down into H2O and CO2 after scaffolds sintering. The appearance of the scaffold constructed by 3D printing technology closely matched the computer design. The incorporation of Sn into β-TCP improved the compressive strength of the scaffold. Moreover, the Sn@TCP scaffold retained the inherently good biocompatibility of β-TCP and exhibited better osteoinduction capability than pure β-TCP scaffolds. Notably, the osteoinduction ability of Sn@TCP scaffolds were dependent on the Sn content. In conclusion, the 3D printing of Sn@TCP scaffolds with enhanced mechanical properties and osteoblast-inducing activity show great promise as scaffold materials in bone tissue engineering applications.

Physio-mechanical and Biological Effects Due to Surface Area Modifications of 3D Printed β-tri- calcium phosphate: An In Vitro Study

Bone defects are associated with trauma, congenital disorders, non-unions, or infections following surgical procedures. Defects which are unable to heal spontaneously are categorized as “critical sized” and are commonly treated using bone grafts in an effort to facilitate bone regeneration and stabilization. Grafting materials can be either natural or synthetic, each having their respective advantages and disadvantages. Synthetic bone grafts are favored due to their ability to be tailored to exhibit desired properties and geometric configurations. β-tricalcium phosphate (β-TCP) is a synthetic grafting material that has been widely utilized for regenerative purposes due to its favorable osteoconductive properties. In combination with 3D printing, grafting materials can be further customized with respect to their macro and micro features. One way to customize devices is by using 3D printing and varying the surface area, by varying the internal component measurements. The objective of this study was to compare the effect of porosity and surface area of 3D printed β-TCP scaffolds with different strut diameters and the effect on cell proliferation in vitro. ß-TCP scaffolds were printed using a custom-built 3D direct-write micro printer with syringes equipped with different extrusion tip diameters (fdiameter: 200 µm, 250 µm and 330 µm). After sintering and post processing, scaffolds were subjected to micro-computed tomography (µCT) and a Scanning Electron Microscope (SEM) to evaluate surface area and porosity, respectively. Compressive strength was assessed using a universal testing machine. Cell proliferation was assessed through cellular viability, using human osteoprogenitor cells. The surface area of the scaffolds was found to increase with smaller strut diameters. Statistically significant differences (p<0.05) were detected for cellular proliferation, between the smallest extrusion diameter, 200 μm, and the largest diameter, 330 μm, after 48-, 72-, and 168-hours. No statistical significances were detected (p>0.05) with regards to the mechanical properties between groups. This study demonstrated that a smaller diameter rod yielded a higher surface area resulting in increased levels of cellular proliferation. Therefore, tailoring rod dimensions has the capacity to enhance cellular adhesion and ultimately, proliferation.

Establishment and biological effect evaluation of prevascularized porous β-tricalcium phosphate tissue engineered bone

To evaluate the biological effect on vascularization during bone repair of prevascularized porous β-tricalcium phosphate (β-TCP) tissue engineered bone (hereinafter referred to as prevascularized tissue engineered bone), which was established by co-culture of endothelial progenitor cells (EPCs) and bone marrow mesenchymal stem cells (BMSCs) based on tissue engineering technology. EPCs and BMSCs were isolated from iliac bone marrow of New Zealand white rabbits by density gradient centrifugation and differential adhesion method. The cells were identified by immunophenotypic detection, BMSCs-induced differentiation, and EPCs phagocytosis. After identification, the third-generation cells were selected for subsequent experiments. First, in vitro tubule formation in EPCs/BMSCs direct contact co-culture (EPCs/BMSCs group) was detected by Matrigel tubule formation assay and single EPCs (EPCs group) as control. Then, the prevascularized tissue engineered bone were established by co-culture of EPCs/BMSCs in porous β-TCP scaffolds for 7 days (EPCs/BMSCs group), taking EPCs in porous β-TCP scaffolds as a control (EPCs group). Scanning electron microscopy and laser scanning confocal microscopy were used to observe the adhesion, proliferation, and tube formation of cells. Femoral condyle defect models of 12 New Zealand white rabbits were used for implantation of prevascularized tissue engineered bone as the experimental group ( n=6) and porous β-TCP scaffold as the control group ( n=6). The process of vascularization of β-TCP scaffolds were observed. The numbers, diameter, and area fraction of neovascularization were quantitatively evaluated by Microfill perfusion, Micro-CT scanning, and vascular imaging under fluorescence at 4 and 8 weeks. The isolated cells were BMSCs and EPCs through identification. EPCs/BMSCs co-culture gradually formed tubular structure. The number of tubules and branches, and the total length of tubules formed in the EPCs/BMSCs group were significantly more than those in the EPCs group on Matrigel ( P<0.05) after 6 hours. After implanting and culturing in porous β-TCP scaffold for 7 days, EPCs formed cell membrane structure and attached to the material in EPCs group, and the cells attached more tightly, cell layers were thicker, the number of cells and the formation of tubular structures were significantly more in the EPCs/BMSCs group than in the EPCs group. At 4 weeks after implantation, neovascularization was observed in both groups. At 8 weeks, remodeling of neovascularization occurred in both groups. The number, diameter, and area fraction of neovascularization in the experimental group were higher than those in the control group ( P<0.05), except for area fraction at 4 weeks after implantation ( P>0.05). The prevascularized tissue engineered bone based on direct contact co-culture of BMSCs and EPCs can significantly promote the early vascularization process during bone defects repair.

3D printed magnesium-doped β-TCP gyroid scaffold with osteogenesis, angiogenesis, immunomodulation properties and bone regeneration capability in vivo

Bioceramics have been used in orthopedic surgery for several years. Magnesium (Mg) is an essential element in bone tissue and plays an important role in bone metabolism. Mg-doped bioceramics has attracted the attention of researchers recently. However, the optimal doping amount of Mg in β-TCP and the immunomodulatory property of Mg-doped β-TCP (Mg-TCP) have not been determined yet. In this study, β-TCP scaffolds doped with different contents of magnesium oxide (0wt%, 1wt%, 3wt%, and 5wt%) with gyroid structure were printed by digital light processing (DLP) method, and the physicochemical and biological functions were then investigated. Mg-doping improved the physicochemical properties of the β-TCP scaffolds. In vitro experiments confirmed that the doping of Mg in β-TCP scaffolds promoted the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and angiogenic differentiation of endothelial progenitor cells (EPCs), where the 5Mg-TCP has the optimal properties when using the "one cell type" method. It was also found that all Mg-TCP facilitated the polarization of RAW264.7 cells to the M2 phenotype, especially the 3Mg-TCP. However, 3Mg-TCP displayed the optimal osteogenic and angiogenic potential when using a "multiple cell type" method, which referred to culturing the BMSCs or EPCs in the macrophage-conditioned medium. Finally, the in vivo experiments were conducted and the results confirmed that the 3Mg-TCP scaffolds possessed the satisfying bone defect repair capability both after 6 and 12weeks of implantation. This study suggests that 3Mg-TCP scaffolds provide the optimal biological performance and thus have the potential for clinical translation.

Impact of Fluid Dynamics on the Viability and Differentiation Capacity of 3D-Cultured Jaw Periosteal Cells

Perfused bioreactor systems are considered to be a promising approach for the 3D culturing of stem cells by improving the quality of the tissue-engineered grafts in terms of better cell proliferation and deeper penetration of used scaffold materials. Our study aims to establish an optimal perfusion culture system for jaw periosteal cell (JPC)-seeded scaffolds. For this purpose, we used beta-tricalcium phosphate (β-TCP) scaffolds as a three-dimensional structure for cell growth and osteogenic differentiation. Experimental set-ups of tangential and sigmoidal fluid configurations with medium flow rates of 100 and 200 µL/min were applied within the perfusion system. Cell metabolic activities of 3D-cultured JPCs under dynamic conditions with flow rates of 100 and 200 µL/min were increased in the tendency after 1, and 3 days of culture, and were significantly increased after 5 days. Significantly higher cell densities were detected under the four perfused conditions compared to the static condition at day 5. However, cell metabolic and proliferation activity under dynamic conditions showed flow rate independency in our study. In this study, dynamic conditions increased the expression of osteogenic markers (ALPL, COL1A1, RUNX2, and OCN) compared to static conditions and the tangential configuration showed a stronger osteogenic effect than the sigmoidal flow configuration.

3D printing of sponge spicules-inspired flexible bioceramic-based scaffolds

Bioceramics are widely used in bone tissue repair and regeneration due to their desirable biocompatibility and bioactivity. However, the brittleness of bioceramics results in difficulty of surgical operation, which greatly limits their clinical applications. The spicules of the marine sponge Euplectella aspergillum (Ea) possess high flexibility and fracture toughness resulting from concentric layered silica glued by a thin organic layer. Inspired by the unique properties of sponge spicules, flexible bioceramic-based scaffolds with spicule-like concentric layered biomimetic microstructures were constructed by combining two-dimensional (2D) bioceramics and 3D printing. 2D bioceramics could be assembled and aligned by modulating the shear force field in the direct ink writing (DIW) of 3D printing. The prepared spicules-inspired flexible bioceramic-based (SFB) scaffolds differentiated themselves from traditional 3D-printed irregular particles-based bioceramic-based scaffolds as they could be adaptably compressed, cut, folded, rolled and twisted without the occurrence of fracture, significantly breaking through the bottleneck of inherent brittleness of traditional bioceramic scaffolds. In addition, SFB scaffolds showed significantly enhanced in vitro and in vivo bone-forming bioactivity as compared to conventional β-tricalcium phosphate (β-TCP) scaffolds, suggesting that SFB scaffolds combined both of excellent mechanical and bioactive characteristics, which is believed to greatly promote the bioceramic science and their clinical applications.

Beta-tricalcium phosphate enhanced mechanical and biological properties of 3D-printed polyhydroxyalkanoates scaffold for bone tissue engineering

Polyhydroxyalkanoates (PHA) is a naturally degradable polyester with good biocompatibility. However, several disadvantages including poor bioactivity and mechanical properties limit the biomedical application of PHA. To circumvent these drawbacks, PHA needs to be blended with other materials to improve performance. Beta-tricalcium phosphate (β-TCP) has emerged as one of the most promising bone repair materials due to its good biocompatibility, satisfactory mechanical properties, and excellent bone osteoconductivity. In this study, PHA filled with β-TCP in 0 wt%, 5 wt%, 10 wt%, 20 wt%, and 30 wt% of concentrations were produced using a twin-screw extruder. The extruded 3D filaments made with 20% β-TCP exhibited the maximum mechanical properties to manufacture 3D scaffolds for bone tissue engineering. We then prepared the 3D-printed PHA/β-TCP scaffolds by using the fused deposition modeling (FDM) technique. The compressive strength and the shore hardness of the PHA/20%β-TCP scaffold were 36.7 MPa and 81.1 HD. The produced scaffolds presented compressive strength compatible with natural bone. In addition, the scaffolds with a well-controlled design of pore shape and size provided sufficient space for cellular activity. In vitro studies demonstrated that the addition of β-TCP could significantly improve the proliferation, adhesion, and migration of MC3T3-E1 cells in the PHA/β-TCP scaffold. Moreover, the osteogenesis-related genes expression of the PHA/β-TCP scaffold was enhanced compared to the PHA scaffolds. Therefore, the 3D-printed PHA/β-TCP scaffold represents an effective strategy to promote mechanical and biological properties, showing huge potential for bone tissue engineering applications.

3D-Printed β-Tricalcium Phosphate Scaffolds Promote Osteogenic Differentiation of Bone Marrow-Deprived Mesenchymal Stem Cells in an N6-methyladenosineDependent Manner

Bone defect is a serious orthopedic disease which has been studied for a long time. Alternative degradable biomaterials are required for bone repairing and regeneration to address the limitation of autogenous bone. β-tricalcium phosphate (β-TCP) is an alternative material with good cytocompatibility and has been used in bone defect treatment. However, whether β-TCP contributes to osteogenesis of bone marrow stem cells (BMSCs) through N6-methyladenosine (m6A) modification remains unknown. To address this issue, we verified the effects of β-TCP on osteogenesis of BMSCs. We also studied the expression of m6A-related enzymes in BMSCs after β-TCP treatment. Furthermore, the m6A level and stability of Runt-related transcription factor 2 (RUNX2) mRNA were investigated after β-TCP treatment. Finally, rat calvarial defect models were performed to detect expression level of osteogenic factors and m6A-related enzymes after the stimulation of three-dimension (3D)-printed β-TCP scaffolds. We found that β-TCP showed good biocompatibility and was osteoinductive. Meanwhile, methyltransferase-like 3 (METTL3) increased, causing the elevation of m6A level of RUNX2, results in stabler RUNX2 mRNA level. At last, based on the animal experiments, we demonstrated that the increase of RUNX2 and METTL3 levels was induced by β-TCP. These findings suggest that METTL3 increases the m6A level of RUNX2 mRNA after β-TCP induction, contributing to its stability, and the results in vivo also confirmed the osteogenic and bone-repair properties of β-TCP.

In vivo evaluation of tricalcium phosphate scaffold for cranial prosthesis application

Tricalcium phosphate beta phase (β-TCP) is a promising bioceramic that stimulates the formation of bone tissue due to its chemical composition, which is similar to those of bones and teeth, showing promising osseointegration and osteoinduction results. Its main applications are in the regeneration and stimulation of cortical and trabecular bone tissue. β-TCP can be used in the human body in different ways: scaffolds, coatings, or granules. In the present investigation, we analyzed the implant bioactivity effects of a β-TCP scaffold in the skull at different timeslots. The scaffolds were characterized physically and chemically before being implanted in vivo. The authors evaluated possible behavioral consequences in cognitive and motor tests on Wistar rats during the time when the tricalcium phosphate scaffolds were reabsorbed, which stimulated the bone regeneration on the cranial defect. The results of the β-TCP scaffolds as skull implant in rats showed an excellent hydroxyapatite (HA) precipitation on the implant's surface, stimulating new bone formation in the defect zone without affecting the motor and cognitive abilities of the animals' during the six weeks after implantation.