Bone Tissue Engineering Research Articles

Structural and biological properties of hydroxyfluorapatite containing sodium and potassium and substituted with carbonates bioceramics for bone tissue engineering

Structural and biological properties of hydroxyfluorapatite containing sodium and potassium and substituted with carbonates bioceramics for bone tissue engineering

Preventing bacterial adhesion on scaffolds for bone tissue engineering

Bone implant infection constitutes a major sanitary concern which is associated to high morbidity and health costs. This manuscript focused on overviewing the main research efforts committed up to date to develop innovative alternatives to conventional treatments, such as those with antibiotics. These strategies mainly rely on chemical modifications of the surface of biomaterials, such as providing it of zwitterionic nature, and tailoring the nanostructure surface of metal implants. These surface modifications have successfully allowed inhibition of bacterial adhesion, which is the first step to implant infection, and preventing long-term biofilm formation compared to pristine materials. These strategies could be easily applied to provide three-dimensional (3D) scaffolds based on bioceramics and metals, of which its manufacture using rapid prototyping techniques was reviewed. This opens the gates for the design and development of advanced 3D scaffolds for bone tissue engineering to prevent bone implant infections.

3D Printed PLA Porous Scaffolds with Engineered Cell Size and Porosity Promote the Effectiveness of the Kelvin Model for Bone Tissue Engineering

AbstractIn this study, the aim is to investigate the effect of engineering the cell size and porosity of 3D‐printed poly lactic acid (PLA) porous scaffolds from the Kelvin model for bone tissue engineering applications. The Kelvin model is used as a bone tissue scaffold with different cell sizes and porosities. PLA, as a biodegradable and biocompatible polymer, is used to fabricate these scaffolds using the FDM technique. A compression test is used to evaluate the mechanical properties of scaffolds. The MTT assay has been used to investigate cell viability. For osteogenic differentiation studies, ALP activity and ARS assays are used. Increasing the porosity reduces the mechanical properties of the scaffold. While increasing the cell size at constant porosity increases the Young's modulus and yield stress in the samples, it is also observed that, in high porosities, the increase in cell size weakens the mechanical properties. Also, Kelvin model scaffolds help the proliferation and osteogenic differentiation of cells and have no toxic effect. It is demonstrated that this approach promotes the effectiveness of the Kelvin architecture for bone tissue engineering. As a result, designing the most suitable model based on cell size and porosity for the treatment process in the targeted area could be promising.

Nanoclay-Composite Hydrogels for Bone Tissue Engineering.

Nanoclay-composite hydrogels represent a promising avenue for advancing bone tissue engineering. Traditional hydrogels face challenges in providing mechanical strength, biocompatibility, and bioactivity necessary for successful bone regeneration. The incorporation of nanoclay into hydrogel matrices offers a potential unique solution to these challenges. This review provides a comprehensive overview of the fabrication, physico-chemical/biological performance, and applications of nanoclay-composite hydrogels in bone tissue engineering. Various fabrication techniques, including in situ polymerization, physical blending, and 3D printing, are discussed. In vitro and in vivo studies evaluating biocompatibility and bioactivity have demonstrated the potential of these hydrogels for promoting cell adhesion, proliferation, and differentiation. Their applications in bone defect repair, osteochondral tissue engineering and drug delivery are also explored. Despite their potential in bone tissue engineering, nanoclay-composite hydrogels face challenges such as optimal dispersion, scalability, biocompatibility, long-term stability, regulatory approval, and integration with emerging technologies to achieve clinical application. Future research directions need to focus on refining fabrication techniques, enhancing understanding of biological interactions, and advancing towards clinical translation and commercialization. Overall, nanoclay-composite hydrogels offer exciting opportunities for improving bone regeneration strategies.

Bone Regeneration of Rat Critical-Sized Calvarial Defects by the Combination of Photobiomodulation and Adipose-Derived Mesenchymal Stem Cells.

Introduction: This study explored the synergistic effects of low-level laser therapy (LLLT) and adipose-derived stem cells (ADSCs) on cranial bone regeneration in rats, addressing the limitations of autogenous grafts and advancing bone tissue engineering with innovative photobiomodulation (PBM) applications. Methods: Sixty Wistar rats were allocated to 5 separate groups randomly; (1) natural bovine bone mineral (NBBM); (2) NBBM+LLLT; (3) NBBM+allogenic ADSCs; (4) NBBM+allogenic ADSCs+LLLT; (5) Only defects. 8-mm calvarial defects were made in each rat in the surgical procedure. A diode laser was applied with the following parameters (continuous mode, power of 100mW, wavelength of 808nm, and 4 J/cm2 energy density) immediately after the procedure and every other day. Bone samples were obtained and assessed histomorphometrically and histologically after staining with hematoxylin and eosin (H&E). Results: Different volumes of bony material were observed in two weeks; 2.94%±1.00 in group 1, 5.1%±1.92 in group 2, 7.11%±2.82 in group 3, 7.34%±2.31 in group 4, and 2.01%±0.83 in group 5 (P<0.05). On the other hand, foreign body residuals were up by 23% in the groups with scaffolding by the end of 2 weeks. Four weeks of observation led to 6.74 %±1.95, 13.24%±1.98, 15.76%±1.19, 15.92%±3.4, and 3.11%±1.00 bone formation in groups 1 to 5, respectively (P<0.05). Generally, the difference between groups 2-4 was not statistically significant based on different types of bone and the extent of inflammation. Conclusion: Bearing in mind the limitations of our research, it was demonstrated that ADSCs in combination with PBM have promising effects on bone tissue regeneration in sizeable bony defects. Furthermore, this study also showed that PBM usage improved the newly regenerated bone quality.

In vitro investigations on the effects of graphene and graphene oxide on polycaprolactone bone tissue engineering scaffolds

Polycaprolactone (PCL) scaffolds that are produced through additive manufacturing are one of the most researched bone tissue engineering structures in the field. Due to the intrinsic limitations of PCL, carbon nanomaterials are often investigated to reinforce the PCL scaffolds. Despite several studies that have been conducted on carbon nanomaterials, such as graphene (G) and graphene oxide (GO), certain challenges remain in terms of the precise design of the biological and nonbiological properties of the scaffolds. This paper addresses this limitation by investigating both the nonbiological (element composition, surface, degradation, and thermal and mechanical properties) and biological characteristics of carbon nanomaterial-reinforced PCL scaffolds for bone tissue engineering applications. Results showed that the incorporation of G and GO increased surface properties (reduced modulus and wettability), material crystallinity, crystallization temperature, and degradation rate. However, the variations in compressive modulus, strength, surface hardness, and cell metabolic activity strongly depended on the type of reinforcement. Finally, a series of phenomenological models were developed based on experimental results to describe the variations of scaffold’s weight, fiber diameter, porosity, and mechanical properties as functions of degradation time and carbon nanomaterial concentrations. The results presented in this paper enable the design of three-dimensional (3D) bone scaffolds with tuned properties by adjusting the type and concentration of different functional fillers.Graphic abstract

Boron substitution in silicate bioactive glass scaffolds to enhance bone differentiation and regeneration

Commercially available bioactive glasses (BAGs) are exclusively used in powder form, due to their tendency to crystallize. Silicate BAG 1393 was developed to allow fiber drawing and scaffold sintering, but its slow degradation limits its potential. To enable scaffold manufacturing while maintaining glass dissolution rate close to that of commercially available BAGs, the borosilicate glass 1393B20 was developed. This study investigates the potential of 1393B20 scaffolds to support bone regeneration and mineralization in vitro and in vivo, in comparison to silicate 1393. Both scaffolds supported human adipose stem cells proliferation, either in direct contact for the 1393, or mainly around for the 1393B20. Similarly, both BAGs induced osteogenesis and angiogenesis in vitro, with a better pro-angiogenic influence of the 1393B20. In addition, these scaffolds supported bone regeneration and osteoclast/osteoblast activity in vivo in critical-sized rat calvarial defect. Nevertheless, mineralization and collagen formation were significantly enhanced for the 1393B20, at 3-months post-implantation, assigned to faster and more complete dissolution of the scaffolds. Thus, 1393B20 demonstrates greater promise for bone tissue engineering certainly due to its time-controlled release of boron and silicon. Statement of significanceBioactive glasses (BAGs) show great promise in bone tissue engineering as they effectively bond with bone tissue, fostering integration and regeneration. Silicate BAG 1393 was developed to allow fiber drawing and scaffold sintering, but its slow degradation limits its potential. To enable scaffold manufacturing while maintaining glass dissolution rate close to that of commercially available BAGs, the borosilicate glass 1393B20 was developed. Both BAGs induced osteogenesis and angiogenesis in vitro, with a better pro-angiogenic influence of the 1393B20. The presence of boron in the 1393B20 enhanced mineralization and collagen formation in vivo compared to 1393, probably due to its faster dissolution rate. Here, 1393B20 demonstrated greater promise for bone tissue engineering compared to the well-known 1393 BAG.

Synthesis and characterization of injectable thermosensitive hydrogel based on Pluronic-grafted silk fibroin copolymer containing hydroxyapatite nanoparticles as potential for bone tissue engineering

Injectable hydrogels are promising for bone tissue engineering due to their minimally invasive application and adaptability to irregular defects. This study presents the development of pluronic grafted silk fibroin (PF-127-g-SF), a temperature-sensitive graft copolymer synthesized from SF and modified PF-127 via a carbodiimide coupling reaction. The PF-127-g-SF copolymer exhibited a higher sol-gel transition temperature (34 °C at 16 % w/v) compared to PF-127 (23 °C), making it suitable for injectable applications. It also showed improved flexibility and strength, with a yielding point increase from <10 % to nearly 30 %. Unlike PF-127 gel, which degrades within 72 h in aqueous media, the PF-127-g-SF copolymer maintained a stable gel structure for over two weeks due to its robust crosslinked hydrogel network. Incorporating hydroxyapatite nanoparticles (n-HA) into the hydrogel reduced pore size and decreased swelling and degradation rates, extending structural stability to four weeks. Increasing n-HA concentration from 0 % to 20 % reduced porosity from 80 % to 66 %. Rheological studies indicated that n-HA enhanced the scaffold's strength and mechanical properties without altering gelation temperature. Cellular studies with MG-63 cells showed that n-HA concentration influenced cell viability and mineralization, highlighting the scaffold's potential in bone tissue engineering.

Effect of unit cell rotation on mechanical performance of selective laser melted Gyroid structures for bone tissue engineering

Gyroid lattice structures, inspired by the natural interconnectivity of human bone, show promise in bone tissue engineering (BTE). This study explores the mechanical properties, and failure mechanisms of selective laser melted (SLM) Gyroid Ti6Al4V lattice structures with varying unit cell rotation angles (RA) of 0∘ (G0), 30∘ (G30), and 60∘ (G60) around the x-axis. The study assesses their mechanical and energy absorption properties through quasi-static compression testing and elastoplastic finite element (FE) analysis. Despite similar relative densities for all designed lattices, the RA variation significantly impacts mechanical performance. G30 exhibits superior load-bearing capacity, with higher modulus of elasticity, yield strength, ultimate strength, and plateau stress. It also has the highest energy absorption capacity, while G60 shows the highest surface area (SA) and surface area-to-volume ratio (SA/VR). These findings highlight the potential of Gyroid lattices for bone replacements due to their tunable mechanical and biological responses.

A multi-scale porous scaffold fabricated by a combined additive manufacturing and chemical etching process for bone tissue engineering

It is critical to develop a fabrication technology for precisely controlling an interconnected porous structure of scaffolds to mimic the native bone microenvironment. In this work, a novel combined process of additive manufacturing (AM) and chemical etching was developed to fabricate graphene oxide/poly(L-lactic acid) (GO/PLLA) scaffolds with multiscale porous structure. Specially, AM was used to fabricate an interconnected porous network with pore sizes of hundreds of microns. And the chemical etching in sodium hydroxide solution constructed pores with several microns or even smaller on scaffolds surface. The degradation period of the scaffolds was adjustable via controlling the size and quantity of pores. Moreover, the scaffolds exhibited surprising bioactivity after chemical etching, which was ascribed to the formed polar groups on scaffolds surfaces. Furthermore, GO improved the mechanical strength of the scaffolds.

Incorporation of azithromycin into akermanite-monticellite nanocomposite scaffolds: Preparation, biological properties, and drug release characteristics

Bioceramics composed of calcium and magnesium silicates have garnered increasing attention for the development of porous scaffolds in bone tissue engineering (BTE). This heightened interest is primarily attributed to their remarkable bioactivity and their capacity to form strong bonds with hard tissue. Fabricating nanocomposite scaffolds is a recognized approach for improving the characteristics of scaffolds used in BTE. This research investigates the mechanical and biological properties, antibacterial activity, and drug-release characteristics of scaffolds composed of akermanite (AKT), monticellite (MON), and monticellite-akermanite (MON-AKT). These scaffolds were fabricated utilizing the space holder process. Additionally, the in vitro drug release profile and antimicrobial activity of Azithromycin (AZT)-loaded MON-AKT composite scaffolds were investigated. The findings showed that the Mon-15 wt% AKT nanocomposite scaffold had the highest density, the smallest grain and micropore sizes, and the lowest porosity. In contrast, integrating AKT into MON-based composite scaffolds resulted in materials characterized by high mechanical strength and stability within physiological environments. The MON-AKT nanocomposite scaffolds exhibited cytocompatibility and demonstrated a high level of alkaline phosphatase (ALP) activity in osteogenic studies. Furthermore, antimicrobial activity assessments revealed that the AZT-encapsulated MON-AKT composite scaffolds effectively inhibited the growth of both S. aureus and E. coli bacteria. The outcomes showed that the antibacterial efficacy of the scaffold depends on both the amount of AZT and the type of bacteria. Overall, MON-AKT/3AZT scaffolds exhibited significantly superior bacterial inhibition compared to other scaffolds, making it a promising option for treating bone tissue defects.

Comparison of hydroxyapatite and honeycomb micro-structure in bone tissue engineering using electrospun beads-on-string fibers.

Thick honeycomb-like electrospun scaffold with nanoparticles of hydroxyapatite (nHA) recently demonstrated its potential to promote proliferation and differentiation of a murine embryonic cell line (C3H10T1/2) to osteoblasts. In order to distinguish the respective effects of the structure and the composition on cell differentiation, beads-on-string fibers were used to manufacture thick honeycomb-like scaffolds without nHA. Mechanical and biological impacts of those beads-on string fibers were evaluated. Uniaxial tensile test showed that beads-on-string fibers decreased the Young Modulus and maximal stress but kept them appropriate for tissue engineering. C3H10T1/2 were seeded and cultured for 6 days on the scaffolds without any growth factors. Viability assays revealed the biocompatibility of the beads-on-string scaffolds, with adequate cells-materials interactions observed by confocal microscopy. Alkaline phosphatase staining was performed at day 6 in order to compare the early differentiation of cells to bone fate. The measure of stained area and intensity confirmed the beneficial effect of both honeycomb structure and nHA, independently. Finally, we showed that honeycomb-like electrospun scaffolds could be relevant candidates for promoting bone fate to cells in the absence of nHA. It offers an easier and faster manufacture process, in particular in bone-interface tissue engineering, permitting to avoid the dispersion of nHA and their interaction with the other cells.

3D-printed titanium scaffolds loaded with gelatin hydrogel containing strontium-doped silver nanoparticles promote osteoblast differentiation and antibacterial activity for bone tissue engineering.

Bone tissue engineering offers a promising alternative to stimulate the regeneration of damaged tissue, overcoming the limitations of conventional autografts and allografts. Recently, titanium alloy (Ti) implants have garnered significant attention for treating critical-sized bone defects, especially with the advancement of 3D printing technology. Although Ti alloys have impressive versatility, their lack of cellular adhesion, osteogenic and antibacterial properties are significant factors that contribute to their failure. Hence, to overcome these obstacles, this study aimed to incorporate osteoinductive and antibacterial cue-loaded hydrogels into 3D-printed Ti (3D-Ti) scaffolds. 3D-Ti scaffolds were synthesized using the direct metal laser sintering method and loaded with a gelatin (Gel) hydrogel containing strontium-doped silver nanoparticles (Sr-Ag NPs). Compared with Ag NPs, Sr-doped Ag NPs increased the expression of Runx2 mRNA, which is a key bone transcription factor. We subjected the bioactive 3D-hybrid scaffolds (3D-Ti/Gel/Sr-Ag NPs) to physicochemical and material characterization, followed by cytocompatibility and osteogenic evaluation. The microporous and macroporous topographies of the scaffolds with Sr-Ag NPs showed increased Runx2 expression and matrix mineralization, with potent antibacterial properties. Therefore, the 3D-Ti scaffolds incorporated with Sr-Ag NP-loaded Gel hydrogels favored osteoblast differentiation and antibacterial activity, indicating their potential for orthopedic applications.

Production of platelet-rich plasma (PRP)-enriched scaffolds for bone tissue regeneration with 3D printing technology

Bone disorders signify diverse abnormalities in the structure, development, and functions of bone tissue in the human body, with a significant correlation to ageing, insufficient physical activity, and escalating obesity. Recent advancements in bone tissue engineering aim to enhance bone tissue formation through the use of biomaterials, growth factors, and cells. The present study focuses on the fabrication and characterisation of scaffolds with a composition of gelatin (GEL) / sodium alginate (SA) / hydroxyapatite (HA) / platelet-rich plasma (PRP) using the 3D printing process. The inclusion of PRP, derived from blood, is of particular interest due to its potential to enhance bone regeneration through various growth factors. Scanning electron microscope (SEM) analysis revealed average pore sizes ranging from 481.50 ± 7.65 to 623.96 ± 11.54 µm. SEM images also showed that scaffold surfaces became smooth as the concentration of PRP increased. The mechanical test results demonstrated that as the PRP increased, the compressive strength decreased. When the swelling and degradation behaviours of scaffolds were examined, it was observed that GEL/SA/HA/3PRP scaffolds exhibited approximately 200 % swelling capability until the 4th day. GEL/SA/HA scaffolds showed a degradation behaviour about 70 % higher compared to other groups. A controlled release profile of PRP was maintained up to the 144th, 216th, and 240th hours from the scaffolds. According to the highest correlation coefficients (R2) in the release kinetics of scaffolds, GEL/SA/HA/0.5PRP and GEL/SA/HA/1PRP scaffolds were explained by the first-order model. In contrast, the GEL/SA/HA/3PRP scaffold was described using the Korsmeyer–Peppas model. The MTT analysis conducted with osteoblast cells showed that scaffolds did not demonstrate any toxic effects and facilitated cell adhesion by inducing the formation of extensions. These findings underscore the potential of PRP-incorporated GEL/SA/HA composites as a promising approach for bone tissue engineering, offering significant advancements in the treatment of bone disorders. This could lead to more effective treatments for bone disorders and injuries, reducing the need for more invasive procedures and improving patient recovery times.

Material extrusion additive manufacturing of graphene oxide reinforced 13–93B1 bioactive glass scaffolds for bone tissue engineering applications

Graphene-reinforced bioactive glass scaffolds have gained significant attention in the field of bone tissue engineering due to their unique combination of mechanical strength, bioactivity, and electrical conductivity. Additive manufacturing techniques, such as 3D printing, provide a versatile platform for fabricating these scaffolds with precise control over their architecture and composition. Consequently, in this work, we have fabricated graphene oxide (GO)-reinforced 13–93B1 bioactive glass scaffold using the material extrusion-based additive manufacturing. A Pluronic F-127-based ink was prepared for scaffold fabrication, and its rheological properties were assessed for shear thinning behaviour, structural support, and recovery. Further, the fabricated scaffolds were characterized using micro-computed tomography, scanning electron microscopy, and energy dispersive x-ray spectroscopy. Additionally, computational fluid dynamics simulations with Dulbecco’s modified eagle medium and blood were performed to evaluate the perfusion kinetics of the scaffolds. The inclusion of GO enhanced the compressive strength of the fabricated scaffolds by ∼202 %. The morphological characterization based on micro-computed tomography showed that additively manufactured scaffolds have appropriate porosity, pore size, pore throat size, and interconnectivity for bone tissue engineering. The synergy with surrounding muscle tissue is also crucial for scaffold-guided bone regeneration. A scaffold supporting the muscle growth around the bone will be able to promote bone-muscle crosstalk and, in turn, bone regeneration. The live-dead assay results showed no cytotoxicity towards C2C12 mouse myoblast cells. Also, cell adhesion and cell viability results show better cell growth on the nanocomposite scaffolds. Overall, the fabricated scaffolds are found suitable for bone tissue engineering applications.

Zinc phosphate glass microspheres promoted mineralization and expression of BMP2 in MC3T3-E1 cells.

Degradable phosphate glasses have shown favorable properties for tissue engineering. By changing the composition of the glasses, the degradation rate, and ion release are controllable. Zinc oxide can function as a glass network modifier and has been shown to play a positive role in bone formation. Also, phosphate glasses can easily be processed into microspheres, which can be used as microcarriers. This study aims to develop zinc phosphate glasses microspheres and explore the optimized size and composition for applications in bone tissue engineering. Zinc-titanium-calcium-sodium phosphate glasses with 0, 1, 3, 5, or 10 mol % zinc oxide were prepared and processed into microspheres. The smaller microspheres ranged in size from 50 to 106 μm, while the larger ones ranged from 106 to 150 μm. The characteristics of glasses were examined. The osteoblastic cell line MC3T3-E1 was cultured on the surface of microspheres and the cell viability was examined. To evaluate osteogenic differentiation, Alizarin Red S staining, quantitative reverse transcription polymerase chain reaction, and western blot analysis were performed after 14 days. Different sizes of zinc phosphate glass microspheres were successfully made. The glass microspheres with <10 mol % zinc oxide were able to support the adhesion and proliferation of MC3T3-E1 cell lines. The relative gene expression of BMP2 was significantly upregulated in the smaller glass microspheres containing 3 mol % zinc oxide (26-fold, p < .001) and both sizes of microspheres containing 5 mol % zinc oxide (smaller: 27-fold, p < .001; larger: 35-fold, p < .001). Additionally, cluster formation was observed in glass microspheres after 14 days, and the mineralization of MC3T3-E1 cell lines was promoted. Based on these findings, the glass microspheres containing 3-5 mol % of zinc oxide can promote osteogenic differentiation for MC3T3-E1 cells.

Chitosan-incorporated Bioceramic-based Nanomaterials for Localized Release of Therapeutics and Bone Regeneration: An Overview of Recent Advances and Progresses

Abstract: The usage of nanoparticles in tissue engineering applications has increased significantly in the last several years. Functional tissues are developed by regulating cell proliferation, differentiation, and migration on nanostructured scaffolds containing cells. These scaffolds provide an environment that is more structurally supportive than the microarchitecture of natural bone. Given its exceptional properties, such as its osteogenic potential, biocompatibility, and biodegradability, chitosan is a good and promising biomaterial. Unfortunately, chitosan's low mechanical strength makes it unsuitable for load-bearing applications. By mixing chitosan with other biomaterials, this drawback might be mitigated. Bone tissue engineering uses both bioresorbable materials like tricalcium phosphate and bioactive materials like hydroxyapatite and bioglass. Alumina and titanium are examples of bioinert materials that are part of these bioceramics. When produced at nanoscale scales, these materials have a larger surface area and better cell adhesion. This review paper will go into great detail on the bioinert, bioresorbable, and bioactive nanoceramics-reinforced chitosan scaffolds for bone tissue engineering.

GO/Cu Nanosheet-Integrated Hydrogel Platform as a Bioactive and Biocompatible Scaffold for Enhanced Calvarial Bone Regeneration.

The treatment of craniofacial bone defects caused by trauma, tumors, and infectious and degenerative diseases is a significant issue in current clinical practice. Following the rapid development of bone tissue engineering (BTE) in the last decade, bioactive scaffolds coupled with multifunctional properties are in high demand with regard to effective therapy for bone defects. Herein, an innovative bone scaffold consisting of GO/Cu nanoderivatives and GelMA-based organic-inorganic hybrids was reported for repairing full-thickness calvarial bone defect. In this study, motivated by the versatile biological functions of nanomaterials and synthetic hydrogels, copper nanoparticle (CuNP)-decorated graphene oxide (GO) nanosheets (GO/Cu) were combined with methacrylated gelatin (GelMA)-based organic-inorganic hybrids to construct porous bone scaffolds that mimic the extracellular matrix (ECM) of bone tissues by photocrosslinking. The material characterizations, in vitro cytocompatibility, macrophage polarization and osteogenesis of the biohybrid hydrogel scaffolds were investigated, and two different animal models (BALB/c mice and SD rats) were established to further confirm the in vivo neovascularization, macrophage recruitment, biocompatibility, biosafety and bone regenerative potential. We found that GO/Cu-functionalized GelMA/β-TCP hydrogel scaffolds exhibited evidently promoted osteogenic activities, M2 type macrophage polarization, increased secretion of anti-inflammatory factors and excellent cytocompatibility, with favorable surface characteristics and sustainable release of Cu2+. Additionally, improved neovascularization, macrophage recruitment and tissue integration were found in mice implanted with the bioactive hydrogels. More importantly, the observations of microCT reconstruction and histological analysis in a calvarial bone defect model in rats treated with GO/Cu-incorporated hydrogel scaffolds demonstrated significantly increased bone morphometric values and newly formed bone tissues, indicating accelerated bone healing. Taken together, this BTE-based bone repair strategy provides a promising and feasible method for constructing multifunctional GO/Cu nanocomposite-incorporated biohybrid hydrogel scaffolds with facilitated osteogenesis, angiogenesis and immunoregulation in one system, with the optimization of material properties and biosafety, it thereby demonstrates great application potential for correcting craniofacial bone defects in future clinical scenarios.

Evaporation-induced self-assembly of hierarchical zinc silicate hybrid scaffolds for bone tissue engineering: Meso and macro scale porosity design

In this study, zinc silicate hybrid scaffolds with hierarchical meso/macroporous structures were synthesized using evaporation-induced self-assembly and sacrificial foamy templates. F127 triblock copolymer and polyurethane (PU) foam served as templates for mesoporosity and macroporosity, respectively. The scaffolds were calcined at 550, 650, and 750 °C for 2 h to remove the templates and form the crystalline phase. Analytical techniques, including BET, XRD, SEM, FTIR, and STA, were used to study the impact of calcination temperature on the scaffolds' meso-texture and crystalline phase. Results showed that increasing the calcination temperature to 750 °C significantly enhanced the overall crystallinity. The crystallized ZnO content increased from 10.3 wt% to 32 wt%, and the willemite (Zn2SiO4) crystalline phase formed. Willemite content was approximately 3 wt% at 650 °C and 67 wt% at 750 °C, as determined by quantitative XRD analyses via Rietveld refinement. The emergence of the willemite phase adversely impacted the specific surface area, leading to a reduction from 123.18 m2/g to 2.18 m2/g, and compromised meso-texture-related characteristics, indicating a substantial disruption in mesoporosity. The pure willemite sample was also obtained by increasing the calcination temperature up to 1000 °C. Although the sample contained no secondary phase, the specific surface area and total mesopore volume were drastically reduced, indicating the negative effect of high calcination temperature on mesoporosity. In contrast to the mesostructure, the macrostructure of the scaffolds exhibited negligible sensitivity to calcination temperature, maintaining a mean macropore diameter of around 200 μm. Furthermore, the in vitro performance of the scaffolds was evaluated by assessing apatite formation ability, degradability, and cytocompatibility. It was demonstrated that the scaffolds calcined at 750 °C exhibited superior performance in terms of apatite formation and cytocompatibility when cultivated with MG-63 human osteosarcoma cells.

Mechanical, physical, and biological properties of polycaprolactone/ Mg-doped SrFe12O19 nanocomposite scaffolds for bone tissue engineering applications

Nowadays, the utilization of 3D printing has become common in the construction of composite scaffolds. In this research, the co-precipitation method was applied to synthesize pure strontium hexaferrite oxide (SrFe12O19) and magnesium-doped strontium hexaferrite oxide nanoparticles (Mg–SrFe12O19). The synthesized SrFe12O19 nanoparticles had a spherical morphology with a mean size of 60 nm and magnetization (Ms) value of 54.38 emu/g. The Mg–SrFe12O19 also had a spherical morphology with a mean particle size of 46 nm and a magnetization value of 29.89 emu/g. The produced polycaprolactone composite scaffolds containing different amounts (0, 25, 50, and 75 wt%) of reinforcement were fabricated by the 3D robocasting printing method. Based on the results of the compression test, the 25 wt% addition of the pure SrFe12O19 nanoparticles to the polycaprolactone scaffold increased the compressive strength from 2.87 to 11.68 MPa compared to the 100 wt% pure polycaprolactone scaffold. Furthermore, the polycaprolactone nanocomposite scaffold containing 25 wt% Mg–SrFe12O19 also increased the compressive strength compared to the pure polycaprolactone scaffold to 12.94 MPa. Therefore, the 25 wt% reinforced scaffold was chosen as the optimal sample. The contact angles of the pure polycaprolactone SrFe12O19 and Mg– SrFe12O19 reinforced polycaprolactone scaffolds were 77.17, 68.53, and 35.58°, respectively. The values indicate the significant effect of doping magnesium on the wettability. The degradability of the 3D-printed scaffolds containing 25 wt% of the two different reinforcements was evaluated for 28 days immersing in phosphate-buffered saline (PBS) solution. The results revealed that the degradability was higher for the Mg–SrFe12O19 after 28 days compared to the composite scaffold reinforced by pure SrFe12O19 reinforcement. Moreover, it was shown that the bioactivity and cell adhesion of MG63 cells for the scaffolds reinforced by Mg–SrFe12O19 have increased in laboratory conditions. Based on the MTT test, it was observed that both of the scaffolds were non-toxic. The results obtained in this study, suggest that polycaprolactone nanocomposite scaffolds containing 25 wt% of Mg–SrFe12O19, made by the 3D robocasting printing method, are suitable for bone tissue engineering.