Silicate Scaffolds Research Articles

3D-printed porous calcium silicate scaffolds with hydroxyapatite/graphene oxide hybrid coating for guided bone regeneration

Calcium silicate (CS) is a suitable bone repair material due to its bioactivity and biocompatibility. However, its rapid degradation rate and poor cell response affect the bone regeneration process. Surface modification of implants represents a potent strategy for enhancing the performance of biomaterials. Here, we present a novel method combining hydrothermal treatment and chemical grafting to tailor the surface morphology and composition of 3D-printed calcium silicate porous scaffolds. The resulting scaffold surface features hydroxyapatite (HA) nanosheets and graphene oxide (GO) sheet structures. Surface modification significantly reduced the degradation rate of CS scaffolds and decreased the Si ion concentration after PBS immersion, which was more conducive to cell survival. Our findings reveal that the CS/HA/GO scaffold exhibits superior biocompatibility and significantly enhances cell adhesion, proliferation, and osteogenic differentiation. In vivo studies demonstrate that the CS/HA/GO scaffold markedly promotes the regeneration of defective bone tissue. This work highlights the potential of HA and GO coatings on interconnected porous scaffolds to induce bone tissue regeneration, offering a promising strategy for orthopedic applications.

3D-printed polycaprolactone/tricalcium silicate scaffolds modified with decellularized bone ECM-oxidized alginate for bone tissue engineering

The treatment of large craniofacial bone defects requires more advanced and effective strategies than bone grafts since such defects are challenging and cannot heal without intervention. In this regard, 3D printing offers promising solutions through the fabrication of scaffolds with the required shape, porosity, and various biomaterials suitable for specific tissues. In this study, 3D-printed polycaprolactone (PCL)-based scaffolds containing up to 30 % tricalcium silicate (TCS) were fabricated and then modified by incorporation of decellularized bone matrix- oxidized sodium alginate (DBM-OA). The results showed that the addition of 20 % TCS increased compressive modulus by 4.5-fold, yield strength by 12-fold, and toughness by 15-fold compared to pure PCL. In addition, the samples containing TCS revealed the formation of crystalline phases with a Ca/P ratio near that of hydroxyapatite (1.67). Cellular experiment results demonstrated that TCS have improved the biocompatibility of PCL-based scaffolds. On day 7, the scaffolds modified with DBM and 20 % TCS exhibited 8-fold enhancement of ALP activity of placenta-derived mesenchymal stem/stromal cells (P-MSCs) compared to pure PCL scaffolds. The present study's results suggest that the incorporation of TCS and DBM-OA into the PCL-based scaffold improves its mechanical behavior, bioactivity, biocompatibility, and promotes mineralization and early osteogenic activity.

The exosomal secretomes of mesenchymal stem cells extracted via 3D-printed lithium-doped calcium silicate scaffolds promote osteochondral regeneration.

The development of surface modification techniques has brought about a major paradigm shift in the clinical applications of bone tissue regeneration. Biofabrication strategies enable the creation of scaffolds with specific microstructural environments and biological components. Lithium (Li) has been reported to exhibit anti-inflammatory, osteogenic, and chondrogenic properties by promoting several intracellular signaling pathways. Currently, research focuses on fabricating scaffolds with simultaneous dual bioactivities to enhance osteochondral regeneration. In this study, we modified the surface of calcium silicate (CS) scaffolds with Li using a simple immersion technique and evaluated their capabilities for bone regeneration. The results showed that Li ions could be easily coated onto the surfaces of CS scaffolds without affecting the microstructural properties of CS itself. Furthermore, the modifications did not affect the printing capabilities of the CS, and porous scaffolds could be fabricated via extrusion. Moreover, the presence of Li improved the surface roughness and hydrophilicity, thus leading to enhanced secretion of osteochondral-related regeneration factors, such as alkaline phosphatase (ALP), bone sialoprotein (BSP), and collagen II (Col II) proteins. Subsequent in vivo studies, including histological and micro-CT analyses, confirmed that the Li-modified CS scaffolds promoted osteochondral regeneration. The transcriptome analysis suggested that the enhanced osteochondrogenic capabilities of our scaffolds were influenced by paracrine exosomes. We hope this study will inspire further research on osteochondral regeneration.

Strontium-doped calcium silicate scaffolds with enhanced mechanical properties and tunable biodegradability fabricated by vat photopolymerization

 Strontium-doped calcium silicate (SrCS) bioceramics have demonstrated outstanding vasculogenic ability to repair large segmental bone defects, while their poor mechanical properties and rapid degradation rate remain the major obstacles in clinical treatment. Here, we proposed a novel approach to significantly enhance the mechanical properties of SrCS bioceramics with tunable biodegradability using micron barium titanate-based (BTA) powders as a dopant. Biomimetic SrCS-BTA scaffolds with triply periodic minimal surface structures were fabricated by vat photopolymerization. The effects of BTA content on microtopography, mechanical properties, degradability, and bioactivity of composite scaffolds were studied. On the one hand, the BTA greatly increased the maximum densification rate of SrCS ceramics by 84.37%, while the corresponding densification temperature decreased by 95°C. On the other hand, CaTiO3 generated by the reaction of SrCS and BTA intercepted cracks at the grain boundaries, and thus, the mechanical properties were enhanced due to the pinning effect. The SrCS-40BTA scaffold exhibited much higher compressive strength and elastic modulus by 296% compared with the pure SrCS scaffold. The energy absorption of SrCS-40BTA scaffolds was 5.6 times higher than that of the pure SrCS scaffold. In addition, biocompatible SrCS-BTA scaffolds with lower degradation rates can play a supporting role in the process of repair for a longer duration. This work provides a promising strategy to fabricate biomimetic scaffolds with highly enhanced mechanical properties and tunable biodegradability for repairing damaged large segmental bone tissues.

3D-Printed Bioceramic Scaffolds with High Strength and High Precision

Due to the increasing cases of bone damage and bone graft demand, bone-repair technology has great social and economic benefits and the manufacturing of artificial bone implants has become a focus in the domain of regenerative therapy. Considering that the traditional manufacturing process cannot effectively control the overall size of the scaffold, the diameter and shape of micropores, and the interoperability of micropores, 3D printing technology has emerged as a focal point of research within the realm of bone tissue engineering. However, the printing accuracy of extrusion-based biological 3D printing techniques is low. In this research, we utilized three-dimensional printing technology to develop high-precision magnesium-containing silicate (CSi-Mg) scaffolds. The precision of this innovative method was scrutinized and the influence of pore size on scaffold strength was systematically analyzed. Furthermore, the influence of the pore architecture on the sidewalls of these 3D-printed scaffolds was evaluated in terms of mechanical properties. The CSi-Mg scaffold, post a 3-week immersion in a simulated body of fluid, demonstrated a high modulus of elasticity (exceeding 404 MPa) and significant compressive strength (beyond 47 MPa). Furthermore, it exhibited commendable bioactivity and biodegradability. These results suggest that the high-precision 3D-printed CSi-Mg scaffolds hold great promise for addressing challenging bone defect cases.

Developing a teriparatide-loaded calcium silicate biomaterial ink for 3D printing of scaffolds for bone tissue engineering

Critical-size bone defects of complex geometries are challenging to repair, making current approaches less satisfactory. Although calcium silicate (CaS) scaffolds prepared using 3D printing can be promising, these scaffolds are commonly treated with harsh conditions to reinforce their strength, significantly affecting the activity of biomolecules in the scaffolds. In this study, we developed a novel CaS biomaterial ink that did not require harsh post-treatments. It is shown that the printability was significantly improved using polyethylene glycol (PEG) and pluronic F-127 (PF), and the scaffolds showed excellent shape fidelity after printing. Moreover, the scaffolds demonstrated compatible mechanical strength with trabecular bone (∼7.07 MPa vs. 0.6 ∼ 16.8 MPa), and adding teriparatide (TP) in scaffolds could further improve the scaffold's potential for bone tissue engineering.

Preparation and characterization of different micro/nano structures on the surface of bredigite scaffolds

The preparation of controllable micro/nano structures on the surface of the bredigite scaffold is expected to exhibit the same support and osteoconductive capabilities as living bone. However, the hydrophobicity of the white calciμm silicate scaffold surface restricts the adhesion and spreading of osteoblasts. Furthermore, during the degradation process of the bredigite scaffold, the release of Ca2+ results in an alkaline environment around the scaffold, which inhibits the growth of osteoblasts. In this study, the three-dimensional geometry of the Primitive surface in the three-periodic minimal surface with an average curvature of 0 was used as the basis for the scaffold unit cell, and a white hydroxyapatite scaffold was fabricated via photopolymerization-based 3D printing. Nanoparticles, microparticles, and micro-sheet structures with thicknesses of 6 μm, 24 μm, and 42 μm, respectively, were prepared on the surface of the porous scaffold through a hydrothermal reaction. The results of the study indicate that the micro/nano surface did not affect the morphology and mineralization ability of the macroporous scaffold. However, the transition from hydrophobic to hydrophilic resulted in a rougher surface and an increase in compressive strength from 45 to 59–86 MPa, while the adhesion of the micro/nano structures enhanced the scaffold's ductility. In addition, after 8 days of degradation, the pH of the degradation solution decreased from 8.6 to around 7.6, which is more suitable for cell growth in the hμman body. However, there were issues of slow degradation and high P element concentration in the degradation solution for the microscale layer group during the degradation process, so the nanoparticle and microparticle group scaffolds could provide effective support and a suitable environment for bone tissue repair.

3D printing of complicated GelMA-coated Alginate/Tri-calcium silicate scaffold for accelerated bone regeneration

Polymer-based composite scaffolds are an attractive class of biomaterials due to their suitable physical and mechanical performance as well as appropriate biological properties. When such composites contain osteoinductive ceramic nanopowders, it is possible, in principle, to stimulate the seeded cells to differentiate into osteoblasts. However, reproducibly fabricating and developing an appropriate niche for cells' activities in three-dimensional (3D) scaffolds remains a challenge using conventional fabrication techniques. Additive manufacturing provides a new strategy for the fabrication of complex 3D structures.Here, an extrusion-based 3D printing method was used to fabricate the Alginate (Alg)/Tri-calcium silicate (C3S) bone scaffolds. To improve physical and biological attributes, scaffolds were coated with gelatin methacryloyl (GelMA), a biocompatible viscose hydrogel. Conducting a combination of experimental techniques and molecular dynamics simulations, it is found that the composition ratio of Alg/C3S governs intermolecular interactions among the polymer and ceramic, affecting the product performance. Investigating the effects of various C3S amounts in the bioinks, the 90/10 composition ratio of Alg/C3S is known as the optimum content in developed bioinks. Accordingly, the printability of high-viscosity inks is boosted by improved hierarchical interactions among assemblies, which in turn leads to better nanoscale alignment in extruded macroscopic filaments. Conducting multiple tests on specimens, the GelMA-coated Alg/C3S scaffolds (with a composition ratio of 90/10) were shown to have improved mechanical qualities and cell adhesion, spreading, proliferation, and osteogenic differentiation, compared to the bare scaffolds, making them better candidates for further future research. Overall, the in-silico and in vitro studies of GelMA-coated 3D-printed Alg/C3S scaffolds open new aspects for biomaterials aimed at the regeneration of large- and complicated-bone defects through modifying the extrusion-based 3D-printed constructs.

Effects of Fillers on the Hydration Behaviors of Tricalcium Silicate Scaffolds Fabricated by Fused Deposition Modeling

With the development of additive manufacturing technology, many types of materials are being utilized, and methods of printing the materials and characteristics of their hydration properties are being studied in the fields of construction and biotechnology. Tricalcium silicate (C3S), which is used as a cement material or biomaterial, is a representative hydraulic material. In previous research, scaffolds were printed via fused deposition modeling and the deformation properties during the hydration process of the printed scaffold were investigated. C3S, like ceramic materials, requires post-processing such as curing after printing, and volumetric deformation occurs during this process. Deformation information is very important to ensure the numerical value of the final product, as well as to suppress the possibility of deformation. In this study, silica, hydroxyapatite (HA), and alumina were mixed with three types of fillers to print a C3S support, which was then cured through a two-step process. In this process, HA and silica, which have good hydrophilicity, exhibited high strength due to the suppression of scaffold deformation. It was confirmed that the smaller the particle size, the more effective it was in obtaining a stable hydrated print.

Robocasting of multicomponent sol-gel–derived silicate bioactive glass scaffolds for bone tissue engineering

Robocasting is universally recognized as an affordable and reproducible manufacturing strategy to process glass and glass-ceramic materials in the form of highly ordered porous scaffolds for bone tissue engineering (BTE) applications. Nevertheless, while being widely applied to melt-derived bioactive glasses, this technique was seldom implemented with sol-gel materials due to the intrinsic difficulties in producing suitable inks for extrusion and thus, good printing outcomes. The present experimental work describes a new and relatively easy method to manufacture multicomponent sol-gel bioactive silicate scaffolds (oxide system: 47.5SiO2–20CaO–10MgO–10Na2O-10 K2O-2.5P2O5, mol.%) using dried gels as basic material within the ink composition, which allows by-passing intermediate heat treatments that are usually detrimental to the bioactive potential of the material in physiological environment. The scaffolds, exhibiting a total porosity of 81 vol%, were characterized in terms of morphological-compositional features and bioactivity in simulated body fluid (SBF), paying special attention to ion release and surface modifications occurring upon soaking (apatite-forming ability). The compressive strength of the scaffolds (around 5 MPa) was comparable to that of human cancellous bone. Collectively, the results supported the possibility of using robocast sol-gel–derived scaffolds in BTE approaches. Furthermore, a comparison with robocast scaffolds based on a melt-derived glass with the same composition was reported in order to investigate the effect of the synthesis route on the dissolution behavior and morphological features of the final biomaterials.

Effect of polycaprolactone impregnation on the properties of calcium silicate scaffolds fabricated by 3D printing

Calcium silicate (CS) is a suitable substrate for bone tissue engineering because it can provide bioactive ions like Si4+ and Ca2+ to promote bone regeneration. However, the rapid degradation of CS leads to pH problems and does not match the rate of osteogenesis. The 3D printed CS scaffolds were immersed in Polycaprolactone (PCL) solution to obtain PCL-coated scaffolds with improved mechanical and biological properties. Finite element method (FEM) analysis found that PCL impregnation has the effect of stress shielding and defect healing, effectively improving the mechanical properties of CS porous scaffolds. The degradation rate of PCL-coated scaffolds was significantly slowed down in Tris buffer. After 4 weeks of degradation, the compressive strength of PCL-coated scaffolds remained at 23.34 MPa, maintaining reliable mechanical properties. PCL coating significantly reduced the degradation rate of CS scaffolds, and the pH value and Si ion concentration of Dulbecco's modified Eagle's medium (DMEM) soaked by PCL-coated scaffolds were more favorable for cell survival. The results show that PCL-coated scaffolds enhance cell proliferation and osteogenic differentiation, demonstrating the potential applicability of bone tissue engineering applications.

3D printed bioceramic scaffolds: Adjusting pore dimension is beneficial for mandibular bone defects repair.

Bioceramic scaffolds for repairing mandibular bone defects have considerable effects, whereas pore architecture in porous scaffolds on osteogenesis in specific structures is still controversial. Herein 6mol% magnesium-substituted calcium silicate scaffolds were fabricated with similar porosity (∼58%) but different cylindrical pore dimensions (Ø 480, 600, and 720μm) via digital light processing-based three-dimensional (3D) printing technique. The mechanical properties, bioactive ion release, and bio-dissolution of the bioceramic scaffolds were evaluated in vitro, and the facilitation of scaffolds on bone formation was investigated after implanting in vivo. The results showed that as the pore dimension increased, the scaffolds indicated similar surface microstructures, but their compressive strength was enhanced gradually. There was no significant difference in vitro bio-dissolution between the 480 and 600μm groups, whereas the 720μm group showed a much slower dissolution and ion release. Interestingly, the two-dimensional/three-dimensional (2D/3D) micro-CT reconstruction analysis of rabbits' mandibular bone defects model showed that the 600μm group exhibited evidently higher ratio of the newly formed bone volume to total volume (BV/TV) and trabecular number (Tb. N) values and lower ratio of the scaffolds residual volume to total volume (RV/TV) compare to the other two sizes. Furthermore, the histological analysis also revealed a considerably higher new bone ingrowth rate in the 600μm group than the other two groups at 4-12weeks post-implantation. Totally, it is proved from these experimental studies that the DLP-based accurately fabricated calcium (Ca) silicate bioceramic scaffolds with appropriate pore dimensions (i.e., 600μm in pore size) are promising to guide new bone ingrowth and thus accelerate the regeneration and repair of cranial maxillofacial or periodontal bone defects.

The effects of a 3D-printed magnesium-/strontium-doped calcium silicate scaffold on regulation of bone regeneration via dual-stimulation of the AKT and WNT signaling pathways

Numerous studies have demonstrated that calcium silicate (CS) can be doped with various trace metal elements such as strontium (Sr) or magnesium (Mg). These studies have confirmed that such modifications promote bone regeneration. However, the development and emergence of 3D printing have further made it possible to fabricate bone grafts with precise structural designs using multi-bioceramics so as to better suit specific clinical requirements. We fabricated scaffolds using Mg-doped CS as the outer layer with Sr-doped CS in the center. In addition, PCL was used to improve printability of the scaffolds. This enhanced Mg and Sr architecture prevented premature degradation of the scaffolds during immersion while enabling the release of ions in a sustained manner in order to achieve the desired therapeutic goals. Even the capabilities of stem cells were shown to be enhanced when cultured on these scaffolds. Furthermore, the hybrid scaffolds were found to up-regulate the expression of bone-related proteins such as factors leading to differentiation-inducing pathways, including PI3K/Akt, Wnt, and TRPM7. The in vivo performance of the proposed scaffolds was assessed using micro-CT. The histological results revealed that the hybrid scaffolds were able to further enhance bone regeneration as compared to uni-bioceramics. By combining 3D printing, multi-ceramics, and trace metal elements, a novel hybrid scaffold could be fabricated with ease and specifically suited to future bone tissue engineering applications.

Effect of Polycaprolactone Impregnation on the Properties of Calcium Silicate Scaffolds Fabricated by 3d Printing

Effect of Polycaprolactone Impregnation on the Properties of Calcium Silicate Scaffolds Fabricated by 3d Printing

Additive manufacturing of Ca–Mg silicate scaffolds supported by flame-synthesized glass microspheres

Novel manufacturing techniques such as additive manufacturing also referred to as 3D printing hold a critical role in the preparation of novel bioactive three-dimensional glass-ceramic scaffolds. The present paper focuses on the use of Ca–Mg silicates microspheres (Ca2MgSi2O7, i.e. 40 mol% CaO, 20% MgO and 40% SiO2) for the fabrication of 3D structures by additive manufacturing. In the first step, the crystallization of the åkermanite system was avoided, by feeding nearly fully crystallized precursor powders prepared by conventional melt quenching into oxygen-methane (O2/CH4) torch, and solid glass microspheres (SGMs) with diameters bellow 63 μm were prepared. In the second step, the crystallization was utilized to control the viscous flow of SGMs during firing of reticulated scaffolds, obtained by digital light processing (DLP) of the SGMs suspended in a photocurable acrylate binder. The spheroidal shape facilitated a high solid content, up to 77 wt% of the SGMs in the suspension. After burn-out of the organic binder, a fast sintering treatment at 950 °C, for 30 min, led to scaffolds preserving the macro-porosity from 3D printing model (diamond cell lattice) but with well densified struts. The crystallization of 3D scaffolds during the sintering process led to 3D structures with adequate strength-to-density ratio.

Effects of Raster Angle and Material Components on Mechanical Properties of Polyether-Ether-Ketone/Calcium Silicate Scaffolds

Polyetheretherketone (PEEK) was widely used in the fabrication of bone substitutes for its excellent chemical resistance, thermal stability and mechanical properties that were similar to those of natural bone tissue. However, the biological inertness restricted the osseointegration with surrounding bone tissue. In this study, calcium silicate (CS) was introduced to improve the bioactivity of PEEK. The PEEK/CS composites scaffolds with CS contents in gradient were fabricated with different raster angles via fused filament fabrication (FFF). With the CS content ranging from 0 to 40% wt, the crystallinity degree (from 16% to 30%) and surface roughness (from 0.13 ± 0.04 to 0.48 ± 0.062 μm) of PEEK/CS scaffolds was enhanced. Mechanical testing showed that the compressive modulus of the PEEK/CS scaffolds could be tuned in the range of 23.3–541.5 MPa. Under the same printing raster angle, the compressive strength reached the maximum with CS content of 20% wt. The deformation process and failure modes could be adjusted by changing the raster angle. Furthermore, the mapping relationships among the modulus, strength, raster angle and CS content were derived, providing guidance for the selection of printing parameters and the control of mechanical properties.

Therapeutic Effects of the Addition of Fibroblast Growth Factor-2 to Biodegradable Gelatin/Magnesium-Doped Calcium Silicate Hybrid 3D-Printed Scaffold with Enhanced Osteogenic Capabilities for Critical Bone Defect Restoration.

Worldwide, the number of bone fractures due to traumatic and accidental injuries is increasing exponentially. In fact, repairing critical large bone defects remains challenging due to a high risk of delayed union or even nonunion. Among the many bioceramics available for clinical use, calcium silicate-based (CS) bioceramics have gained popularity due to their good bioactivity and ability to stimulate cell behavior. In order to improve the shortcomings of 3D-printed ceramic scaffolds, which do not easily carry growth factors and do not provide good tissue regeneration effects, the aim of this study was to use a gelatin-coated 3D-printed magnesium-doped calcium silicate (MgCS) scaffold with genipin cross-linking for regulating degradation, improving mechanical properties, and enhancing osteogenesis behavior. In addition, we consider the effects of fibroblast growth factor-2 (FGF-2) loaded into an MgCS scaffold with and without gelatin coating. Furthermore, we cultured the human Wharton jelly-derived mesenchymal stem cells (WJMSC) on the scaffolds and observed the biocompatibility, alkaline phosphatase activity, and osteogenic-related markers. Finally, the in vivo performance was assessed using micro-CT and histological data that revealed that the hybrid bioscaffolds were able to further achieve more effective bone tissue regeneration than has been the case in the past. The above results demonstrated that this type of processing had great potential for future clinical applications and studies and can be used as a potential alternative for future bone tissue engineering research, as well as having good potential for clinical applications.

Effect of sodium carbonate addition on the properties of calcium silicate scaffolds fabricated by 3DIP

3D ink printing (3DIP) technology can accurately control the macroscopic size and microstructure of bioceramic scaffolds. However, nonceramic components in the printing ink used in 3DIP severely affect densification, resulting in less desirable mechanical properties of the scaffolds. In this study, a strategy of sintering assisted by a sintering aid (sodium carbonate) was used to prepare calcium silicate (CSi) scaffolds with high strength. The addition of 1% sintering aid to a CSi scaffold sintered at 1100 °C led to an appreciable compressive strength (47.8 MPa) and high elastic modulus (1847 MPa). Moreover, the CSi scaffolds with sintering aids showed better degradation ability and mineralization ability than the CSi scaffolds without sintering aids. It is expected that the method involving strengthening with sintering aids will promote the application of 3DIP bioceramic scaffolds in the repair of bone defects.

Effect of Fe3O4 concentration on 3D gel-printed Fe3O4/CaSiO3 composite scaffolds for bone engineering

In this study, porous calcium silicate (CaSiO3) scaffolds were prepared by 3D gel-printing (3DGP) method and Fe3O4 water-based magnetic fluids (WMFs) were prepared by phacoemulsification compound chemical coprecipitation method. Fe3O4 WMFs were coated on CaSiO3 scaffolds surface to prepare Fe3O4/CaSiO3 composite scaffolds. The effect of WMFs with different Fe3O4 concentrations on porous CaSiO3 scaffolds was studied. The composition and morphological characteristics of porous scaffolds were analyzed by using scanning electron microscope (SEM) and energy dispersive spectrometer (EDS) analysis. The magnetic properties were tested by vibrating sample magnetometer (VSM). The stability of Fe3O4 WMFs coatings and the degradability of composite scaffolds were tested by immersing them in simulated body fluid (SBF). The results show that when Fe3O4 concentration was 5.4% (w/v), the composite scaffolds had the highest saturation magnetization of 69.6 emu/g and the best stability in dynamic SBF. It is obviously that Fe3O4 WMFs coatings can be used for bone tissue engineering scaffolds repairing.

Bioactive alginate/carrageenan/calcium silicate porous scaffolds for bone tissue engineering

Porous bioactive alginate/carrageenan/calcium silicate scaffolds for bone tissue engineering were fabricated. The scaffolds were prepared by dispersing the synthesized calcium silicate in an aqueous solution of alginate and carrageenan at 90 °C. The scaffolds were shaped by freeze-drying and further crosslinked by 0.5, 1.0 and 1.5 M CaCl2 for 60 and 120 min. The scaffolds crosslinked by 1.5 M CaCl2 for 120 min achieved the highest in vitro dimension stability. The formation of hydroxyapatite crystals was observed on the scaffolds surface after soaking in simulated body fluid (SBF) at 37 °C for 7–28 days, indicating in vitro bioactivity of the scaffolds. The presence of calcium silicate could enhance not only the bioactivity, but also the mechanical properties of the scaffolds comparable to the cancellous bone. Moreover, the dimension and mechanical properties of the wet scaffolds could recover to the original after four cycles of mechanical testing at 50 % strain. The scaffolds were nontoxic to human living cells, in which the in vitro drug release behavior of the scaffold using diclofenac as a model drug was suitable for the treatment of acute inflammation after surgery. Therefore, these scaffolds were considered to be the candidate materials for bone replacement.