Materials For Bone Tissue Engineering Research Articles

Electrochemical Mineralization Regulates Hydroxyapatite Deposition of Silk Fibroin Nanofibers for Promoting Osteogenic Differentiation of Human Mesenchymal Stem Cells

ABSTRACTScaffold and stem cells are the key elements in the procedure of bone repair. Bombyx mori silk fibroin (SF) is the proper material for bone tissue engineering due to its biocompatibility and its easy to obtain nanofiber structure. The suitable cell substrate is also an important factor because different cells have different adaptations to composite. For this case, selecting a scaffold that can promote osteogenic differentiation of various cells is crucial and meaningful. In this work, hydroxyapatite (HA) was deposited precisely onto each silk nanofiber by electrochemical mineralization (EC) to form SF/HA. SF/HA can promote the proliferation and osteogenic differentiation of both human bone marrow mesenchymal‐derived stem cells (hMSCs) and human adipose‐derived mesenchymal stem cells (hAMSCs). It also promot the expression of alkaline phosphatase (ALP) and osteogenic differentiation related genes. Western blot analyses show mitogen‐activated protein kinase (MAPK) signal pathway is regulated by SF/HA. Therefore, the study provides a proper method to obtain a good composite SF/HA and it promotes the osteogenic differentiation of both hMSCs and hAMSCs.

Fabrication and Biomedical Application of Alginate Composite Hydrogels in Bone Tissue Engineering: A Review.

Nowadays, as a result of the frequent occurrence of accidental injuries and traumas such as bone damage, the number of people causing bone injuries or fractures is increasing around the world. The design and fabrication of ideal bone tissue engineering (BTE) materials have become a research hotspot in the scientific community, and thus provide a novel path for the treatment of bone diseases. Among the materials used to construct scaffolds in BTE, including metals, bioceramics, bioglasses, biomacromolecules, synthetic organic polymers, etc., natural biopolymers have more advantages against them because they can interact with cells well, causing natural polymers to be widely studied and applied in the field of BTE. In particular, alginate has the advantages of excellent biocompatibility, good biodegradability, non-immunogenicity, non-toxicity, wide sources, low price, and easy gelation, enabling itself to be widely used as a biomaterial. However, pure alginate hydrogel as a BTE scaffold material still has many shortcomings, such as insufficient mechanical properties, easy disintegration of materials in physiological environments, and lack of cell-specific recognition sites, which severely limits its clinical application in BTE. In order to overcome the defects of single alginate hydrogels, researchers prepared alginate composite hydrogels by adding one or more materials to the alginate matrix in a certain proportion to improve their bioapplicability. For this reason, this review will introduce in detail the methods for constructing alginate composite hydrogels, including alginate/polymer composite hydrogels, alginate/bioprotein or polypeptide composite hydrogels, alginate/bioceramic composite hydrogels, alginate/bioceramic composite hydrogels, and alginate/nanoclay composite hydrogels, as well as their biological application trends in BTE scaffold materials, and look forward to their future research direction. These alginate composite hydrogel scaffolds exhibit both unexceptionable mechanical and biochemical properties, which exhibit their high application value in bone tissue repair and regeneration, thus providing a theoretical basis for the development and sustainable application of alginate-based functional biomedical materials.

Preparation and Structure-Property Relationship Study of Piezoelectric-Conductive Composite Polymer Nanofiber Materials for Bone Tissue Engineering.

This study aimed to develop Janus-, cross-network-, and coaxial-structured piezoelectric-conductive polymer nanofibers through electrospinning to mimic the piezoelectricity of bone and facilitate the conduction of electrical signals in bone tissue repair. These nanofibers were constructed using the piezoelectric polymer polyvinylidene fluoride, and the conductive fillers reduced graphene oxide and polypyrrole. The influence of structural features on the electroactivity of the fibers was also explored. The morphology and components of the various structural samples were characterized using SEM, TEM, and FTIR. The electroactivity of the materials was assessed with a quasi-static d33 meter and the four-probe method. The results revealed that the piezoelectric-conductive phases were successfully integrated. The Janus-structured nanofibers demonstrated the best electroactivity, with a piezoelectric constant d33 of 24.5 pC/N and conductivity of 6.78 × 10-2 S/m. The tensile tests and MIP measurements showed that all samples had porosity levels exceeding 70%. The tensile strength of the Janus and cross-network structures exceeded that of the periosteum (3-4 MPa), with average pore sizes of 1194.36 and 2264.46 nm, respectively. These properties indicated good mechanical performance, allowing material support while preventing fibroblast invasion. The CCK-8 and ALP tests indicated that the Janus-structured samples were biocompatible and significantly promoted the proliferation of MC3T3-E1 cells.

In vitro biocompatibility and stem cell regenerative assessment of hollow hydroxyapatite spheres deposited wollastonite/Ca2P6O17/TCP/doped-wollastonite scaffolds

This research examines the biological behavior of multilayer ceramic-glass scaffolds for bone tissue engineering. The scaffolds consist of a high-strength core of wollastonite and P6 (Ca2P6O17), with a sequential coating of TCP and wollastonite doped with Na+, K+ and Mg2+ ions to confer bioactivity to the system. Two formulations were investigated: a magnesium-deficient one, which served as a microstructure control, and a magnesium-rich one in which hollow hydroxyapatite (HA) spheres precipitated after one week of immersion in simulated body fluid (SBF). These formulations are denoted as C-Scaffold and HA-Scaffold, respectively. Further exploration and comparison of the physiological effects of both scaffolds is necessary to fully understand their potential as drug release agents and associated therapeutic advantages. The study considered aspects such as protein adhesion, cell morphology, and the inflammatory response. The results showed that the HA-Scaffold had over 34 % more plasma protein adhesion compared to the C-Scaffold. Similarly, cell proliferation assays showed increased cell growth in both HFF-1 fibroblasts and mesenchymal stem cells (MSCs) treated with HA-Scaffold compared to C-Scaffold. This may be attributed to the higher protein adsorption, which acts as a facilitator for cell adhesion. Three-dimensional photography from computerized microtomography confirmed that the scaffolds have a similar morphology and porosity to the intricate structure of trabecular bone in the humeral head. The study shows that the scaffolds made of Wollastonite/Ca2P6O17/TCP/Doped-Wollastonite with HA hollow spheres are highly suitable for use in this bone segment due to their overall porosity of over 86 % and approximate mechanical strength of 1.6 MPa. These findings demonstrate the biocompatibility and enhanced biological properties of the scaffolds, making them an excellent material for bone tissue engineering.

An intelligent controlled release hydrogel with photothermal effect for the treatment of bone defects

To solve the difficulty of natural healing in patients with severe bone defects, and the challenges of insufficient autologous bone graft material and large incisions in conventional surgery, we were designed an intelligent co-regulated hydrogel system with NIR response. The goal is to be able to prevent inflammation and bacterial adhesion, thereby promoting repair and development of the bone defect area. The hydrogel system consisted of hyaluronic acid, carboxymethyl chitosan and Pht-loaded PDA-encapsulated ZIF-8 NPs (Pht@ZIF-8@PDA) by Schiff base reaction. Compared to high-temperature photothermal therapy (PTT), which destroyed surrounding healthy cells, the present study was based on a new strategy of mild photothermal therapy (42.5 ± 1.2 °C) and leaded to a better promotion of osteogenesis. NIR activated the photothermal properties of PDA in scaffolds, intelligently released Pht and Zn2+, inhibited Escherichia coli and Staphylococcus aureus (inhibition rate ≥ 96 %), and promoted bone regeneration in the rat model. Meanwhile, it could promote bone regeneration by upregulating osteogenic factors and stimulating the expression of proteins related to BMP/SMAD signaling pathway. The research highlights the potential applications of synergistically modulated hydrogels, which opens up a new application path for this material in bone tissue engineering, regenerative medicine, and other related fields.

Bio-Printing of Materials for Bone Tissue Engineering

Abstract: The complicated internal mechanical and structural qualities of normal bone tissue still prevent the development of effective therapeutic procedures for major bone lesions. It is still difficult to use tissue engineering to return damaged bones back to how they were originally intended. Due to recent advances in 3D printing, together with the introduction of new materials and technological assistance, the basis for BTE has been established. Biological 3D biomaterials have cells inside them, which allows for the creation of structures that mimic real tissues. Microextrusion, inkjet, and laser-assisted bioprinting are the three primary methods used in 3D bioprinting manufacturing. Hydrogels packed with cells, growth hormones, and bioactive ceramics are among the bioinks utilized in bone bioprinting. With the use of magnetic resonance imaging or computed tomography scanning, 3D printing offers substantial benefits for tailored treatment by enabling the creation of scaffolds with the right structural qualities, form, and dimensions. Three-dimensional (3D) bioprinting is a cutting-edge technique that has been utilized recently to create multicellular, biomimetic tissues with layers upon layers of intricate tissue microenvironment printing. We approached the use of hydrogels with great strength in 3D printing for BTE with an emphasis on first providing a thorough study about the development of 3D printing, printing techniques, and ink selection in this review. A brief prediction on how 3D printing would advance in the future was made.

Bone-inspired AZ31/HA composites with gradient structures and properties for implant applications

The increasing demand for lightweight, damage-resistant, wear-resistant, and corrosion-resistant materials in implant applications has driven the development of novel bone-inspired composites. This study introduces AZ31 (Mg–3Al–1Zn)/hydroxyapatite (HA) composites fabricated using centrifugal freeze casting and pressureless infiltration. The unique formation mechanism and structure-property relationship of these composites were investigated. The gradient distribution of HA particles and the nucleation and growth behavior of ice crystals, influenced by varying solid content in the centrifugal field, resulted in a radially increasing ceramic-layer-to-metal-layer thickness ratio. Consequently, the composites exhibited a gradient distribution of compression strength, hardness, wear resistance, and corrosion resistance. Moreover, the composites showed no cytotoxicity and promoted osteoblast proliferation, making them suitable for bone tissue engineering applications. These findings provide valuable insights for developing new materials in bone tissue engineering and pave the way for future research in this field.

Mixed alkali effect on the mechanical, thermal and biological properties of 58S bioactive glass

Bioactive glasses (BG) containing silicon, phosphorous, and boron have emerged as promising materials for bone tissue engineering. Modifying ions play a multifaceted role in BGs, influencing their structure, properties, and biological activity that depends on their size, charge, and coordination chemistry. This study investigates the addition of sodium borate (Na2B4O7) to the 58S BG and the mixed alkali effect (MAE) resulting from incorporating Na and B ions into the CaO–SiO2–P2O5 system. In specific concentrations, Na+ and Ca2+ ions function as network modifiers, introducing structural disruptions and generating defects that impact the glass transition temperature (Tg), crystallization temperature (Tc), hardness, and sintering characteristics. The physicochemical characterization of BGs confirms the incorporation of Na and B structures. Furthermore, the Na and B doped glass sample with a [Na/Si] ratio of 0.045 exhibits favorable properties for promoting cell attachment, including forming a hydroxyapatite (HAp) layer over the surface, thereby accelerating human placental mesenchymal stem cells (hPMSCs) proliferation. The findings demonstrate the potential of Na2B4O7 incorporation in BG for bone tissue engineering applications.

Osteoblast responsive biosilica-enriched gelatin microfibrillar microenvironments

Engineering of scaffolds for bone regeneration is often inspired by the native extracellular matrix mimicking its composite fibrous structure. In the present study, we used low loadings of diatomite earth (DE) biosilica to improve the bone regeneration potential of gelatin electrospun fibrillar microenvironments. We explored the effect of increasing the DE content from 1 % to 3 % and 5 %, respectively, on the physico-chemical properties of the fibrous scaffolds denoted FG_DE1, FG_DE3, FG_DE5, regarding the aqueous media affinity, stability under simulated physiological conditions, morphology characteristics, and local mechanical properties at the surface. The presence of biosilica generated composite structures with lower swelling degrees and higher stiffness when compared to gelatin fibers. Increasing DE content led to higher Young modulus, while the stability of the protein matrix in PBS, at 37 °C, over 21 was significantly decreased by the presence of diatomite loadings. The best preosteoblast response was obtained for FG_DE3, with enhanced mineralization during the osteogenic differentiation when compared to the control sample without diatomite. 5 % DE in FG_DE5 proved to negatively influence cells' metabolic activity and morphology. Hence, the obtained composite microfibrillar scaffolds might find application as osteoblast-responsive materials for bone tissue engineering.

Enhancement of Bone Tissue Regeneration with Multi-Functional Nanoparticles by Coordination of Immune, Osteogenic, and Angiogenic Responses.

Inorganic nanoparticles are promising materials for bone tissue engineering due to their chemical resemblance to the native bone structure. However, most studies are unable to capture the entirety of the defective environment, providing limited bone regenerative abilities. Hence, this study aims to develop a multifunctional nanoparticle to collectively control the defective bone niche, including immune, angiogenic, and osteogenic systems. The nanoparticles, self-assembled by biomimetic mineralization and tannic acid (TA)-mediated metal-polyphenol network (MPN), are released sustainably after the incorporation within a gelatin cryogel. The released nanoparticles display a reduction in M1 macrophages by means of reactive oxygen species (ROS) elimination. Consequently, osteoclast maturation is also reduced, which is observed by the minimal formation of multinucleated cells (0.4%). Furthermore, the proportion of M2 macrophages, osteogenic differentiation, and angiogenic potential are consistently increased by the effects of magnesium from the nanoparticles. This orchestrated control of multiple systems influences the in vivo vascularized bone regeneration in which 80% of the critical-sized bone defect is regenerated with new bones with mature lamellar structure and arteriole-scale micro-vessels. Altogether, this study emphasizes the importance of the coordinated modulation of immune, osteogenic, and angiogenic systems at the bone defect site for robust bone regeneration.

Silver and magnesium co-doped β-TCP nanoparticles as potential materials for bone tissue engineering

Silver and magnesium co-doped β-TCP nanoparticles as potential materials for bone tissue engineering

Properties of Carbonated Hydroxyapatite-Based Scaffold from Oyster Shells Composited with Honeycomb and Polyethylene Oxide for Bone Tissue Engineering Applications

Scaffold Carbonated Hydroxyapatite/Honeycomb/Polyethylene Oxide (CHA/HCB/PEO) has been obtained by freeze-drying. The bioceramic CHA used in this study was synthesized from oyster shells using precipitation. HCB and PEO were added as reinforcement materials that affect the crystallographic properties of the scaffold. This study aimed to determine the characteristics of the scaffolds for bone tissue engineering. CHA and scaffolds were characterized using Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffractometer (XRD), and Scanning Electron Microscopy (SEM). FTIR spectra and XRD graphs confirmed that the CHA produced was B-type. FTIR spectra of the scaffold showed the presence of HCB and PEO in the scaffold, which means they were homogeneously bound in the scaffold solution. XRD test results show that scaffolds' crystallinity and crystallite size tends to decrease compared to CHA. This was good because they could make cells easier to proliferate. A small-scale pore structure (micropore) was also formed in the scaffold. The porosity and pore size of the scaffold were affected by the concentration of CHA. The presence of the micropores can increase the permeability of the scaffold and facilitate cell migration. Thus, the composition of CHA/HCB/PEO scaffolds can be a good candidate material in bone tissue engineering.

Numerical study on effect of process parameters on material extrusion 3D printing (ME3DP) for porous bone tissue engineering scaffolds

Bone tissue engineering (BTE) holds promise for treating bone defects, and 3D-printed porous scaffolds are pivotal for tissue regeneration. This study explores the impact of material extrusion 3D printing (ME3DP) process parameters (i.e., layer height and extrusion temperature) on 3D-printed porous scaffolds made of polyetherimide (PEI) material for BTE. The intricate relationships shaping scaffold dimensional accuracy, residual stresses, and warpage tendencies were unveiled using comprehensive numerical simulations. Our findings reveal that higher layer heights enhance dimensional stability, especially at lower printing temperatures, reducing maximum displacements. Consequently, lower layer heights lead to higher warpage critical values. This insight is instrumental in optimizing the material extrusion 3D printing (ME3DP) process, vital for precise net shape in BTE applications. Furthermore, residual stresses also revealed a correlation between higher printing temperatures and increased residual stresses. These findings emphasize the necessity of balancing material flow, printing temperature, and layer height for scaffold quality. Therefore, the right balance between precision and mechanical stability is critical for producing high-quality scaffolds that meet BTE requirements.

Simultaneous Regulation of the Mechanical/Osteogenic Capacity of Brushite Calcium Phosphate Cement by Incorporating with Poly(ethylene glycol) Dicarboxylic Acid.

Brushite calcium phosphate cement (brushite CPC) is a prospective bone repair material due to its ideal resorption rates in vivo. However, the undesirable mechanical property and bioactivity limited its availability in clinic application. To address this issue, incorporating polymeric additives has emerged as a viable solution. In this study, poly(ethylene glycol) dicarboxylic acid, PEG(COOH), was synthesized and employed as the polymeric additive. The setting behavior, anti-washout ability, mechanical property, degradation rate, and osteogenic capacity of brushite CPC were regulated by incorporating PEG(COOH). The incorporation of PEG(COOH) with carboxylic acid groups demonstrated a positive effect on both mechanical properties and osteogenic activity in bone repair. This study offers valuable insights and suggests a promising strategy for the development of materials in bone tissue engineering.

3D printed high-precision porous scaffolds prepared by fused deposition modeling induce macrophage polarization to promote bone regeneration

Macrophage-mediated bone immune responses significantly influence the repair of bone defects when utilizing tissue-engineered scaffolds. Notably, the scaffolds’ physical structure critically impacts macrophage polarization. The optimal pore size for facilitating bone repair remains a topic of debate due to the imprecision of traditional methods in controlling scaffold pore dimensions and spatial architecture. In this investigation, we utilized fused deposition modeling (FDM) technology to fabricate high-precision porous polycaprolactone (PCL) scaffolds, aiming to elucidate the impact of pore size on macrophage polarization. We assessed the scaffolds’ mechanical attributes and biocompatibility. Real-time quantitative reverse transcription polymerase chain reaction was used to detect the expression levels of macrophage-related genes, and enzyme linked immunosorbent assay for cytokine secretion levels. In vitro osteogenic capacity was determined through alkaline phosphatase and alizarin red staining. Our findings indicated that macroporous scaffolds enhanced macrophage adhesion and drove their differentiation towards the M2 phenotype. This led to the increased production of anti-inflammatory factors and a reduction in pro-inflammatory agents, highlighting the scaffolds’ immunomodulatory capabilities. Moreover, conditioned media from macrophages cultured on these macroporous scaffolds bolstered the osteogenic differentiation of bone marrow mesenchymal stem cells, exhibiting superior osteogenic differentiation potential. Consequently, FDM-fabricated PCL scaffolds, with precision-controlled pore sizes, present promising prospects as superior materials for bone tissue engineering, leveraging the regulation of macrophage polarization.

Performance of 3D printed porous polyetheretherketone composite scaffolds combined with nano-hydroxyapatite/carbon fiber in bone tissue engineering: a biological evaluation

Polyetheretherketone (PEEK) has been one of the most promising materials in bone tissue engineering in recent years, with characteristics such as biosafety, corrosion resistance, and wear resistance. However, the weak bioactivity of PEEK leads to its poor integration with bone tissues, restricting its application in biomedical fields. This research effectively fabricated composite porous scaffolds using a combination of PEEK, nano-hydroxyapatite (nHA), and carbon fiber (CF) by the process of fused deposition molding (FDM). The experimental study aimed to assess the impact of varying concentrations of nHA and CF on the biological performance of scaffolds. The incorporation of 10% CF has been shown to enhance the overall mechanical characteristics of composite PEEK scaffolds, including increased tensile strength and improved mechanical strength. Additionally, the addition of 20% nHA resulted in a significant increase in the surface roughness of the scaffolds. The high hydrophilicity of the PEEK composite scaffolds facilitated the in vitro inoculation of MC3T3-E1 cells. The findings of the study demonstrated that the inclusion of 20% nHA and 10% CF in the scaffolds resulted in improved cell attachment and proliferation compared to other scaffolds. This suggests that the incorporation of 20% nHA and 10% CF positively influenced the properties of the scaffolds, potentially facilitating bone regeneration. In vitro biocompatibility experiments showed that PEEK composite scaffolds have good biosafety. The investigation on osteoblast differentiation revealed that the intensity of calcium nodule staining intensified, along with an increase in the expression of osteoblast transcription factors and alkaline phosphatase activities. These findings suggest that scaffolds containing 20% nHA and 10% CF have favorable properties for bone induction. Hence, the integration of porous PEEK composite scaffolds with nHA and CF presents a promising avenue for the restoration of bone defects using materials in the field of bone tissue engineering.

Osphronemus goramy scales-derived type 1 Collagen induces RUNX2 and Osteocalcin expression: An in vivo Study

Introduction: Alveolar bone defects need bone augmentation therapies by subtituting with bone material. Gourami (Osphronemus goramy) fish scale comprises type 1 collagen and it has been used as scaffolding material in bone tissue engineering. As alternative bone graft material, the scales have a big potency to promote osteogenesis in periodontal bone defect when autografts are not feasible. This study will analise Runx2 and Osteocalcin expression in wistar rat alveolar bone induced by type 1 collagen derived from gourami fish scale. Methods: 32 male Wistar rats were divided into four groups; control group—7 days (C7), treatment group—7 days (P7), control group—14 days (C14), and treatment group—14 days (P14). The left mandibular incisivus was extracted and the tooth socket was treated with 10mg collagen. The rats were euthanized (at day 7th and 14th) and immunohistochemistry was performed using monoclonal antibodies anti-RUNX2 and anti-osteocalcin. Results: After seven days and 14days, the expression of RUNX2 and osteocalcin in the treatment group increased significantly (p<0.05) compared with the control group. Conclusion: Type 1 collagen from gourami (Osphronemus goramy) fish scales increases RUNX2 and osteocalcin expression as a bone growth marker.

Synthesis of pure and Cd-doped hydroxyapatite for the photo-catalytic degradation of Amoxicillin and Ciprofloxacin: Crystallographic characterization using XRD

Nanosized hydroxyapatite (HAp) is considered an elementary material for artificial bone tissue engineering and the applications are expanding and one of them is the photocatalytic degradation of environmentally hazardous pollutants. In this research, HAp nanoparticles, in pure form and doped with cadmium (Cd), were synthesized using a conventional wet-chemical precipitation method. Additionally, the synthesized samples such as pure and Cd-doped HAp were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and UV–Vis spectroscopy. Moreover, the XRD analysis allowed for a detailed estimation of the crystallite size, assisting in their tailoring of increased photocatalytic activity. The crystallite sizes of the nano-crystallite Hap were evaluated using the Scherrer Method (SM), Williamson-Hall Method (WHM), Size-Strain Plot (SSP), Halder-Wagner Method (HWM), and Sahadat-Scherrer Method (SSM). The models confirmed the formation of nano crystallite hydroxyapatite in pure and doped form. Cadmium doping in HAp showed great photocatalytic activity for example at 150 min, 31 % amoxicillin and 52 % ciprofloxacin were degraded upon 1.5 wt % Cd doping in HAp crystal. The research demonstrated that doping Cd in HAp crystal improved photocatalytic activity towards antibiotic solution (ciprofloxacin and amoxicillin) under all test circumstances.

Effects of aligned PLGA/SrCSH composite scaffolds on in vitro growth and osteogenic differentiation of human mesenchymal stem cells.

Strontium (Sr) has important functions in bone remodeling. Incorporating strontium-doped α-calcium sulfate hemihydrate (SrCSH) into poly(lactic-co-glycolic acid) (PLGA) fibrous scaffolds were expected to increase its bio-activity and provide a potential material for bone tissue engineering. In the present study, Sr-containing aligned PLGA/SrCSH fibrous scaffolds similar to the architecture of natural bone were prepared via wet spinning. CCK-8 assay revealed that Sr-containing scaffolds possessed better bioactivity and supported favorable cell growth effectively. The aligned PLGA/SrCSH fibers exerted a contact effect on cell attachment, inducing regular cell alignment and influencing a series of cell behaviors. Releasing of high concentration Sr from a-PLGA/SrCSH scaffolds could induce high expression levels of BMP-2, increase ALP activity and upregulate RUNX-2 expression, and further promote the expressions of COL-I and OCN and the maximum mineralization. This study demonstrated that Sr and ordered structure in a-PLGA/SrCSH fibrous scaffolds could synergistically enhance the osteogenic differentiation of umbilical cord mesenchymal stem cells (UCMSCs) by regulating cell arrangement and expressions of osteogenic genes.

Magnesium-Doped Nano-Hydroxyapatite/Polyvinyl Alcohol/Chitosan Composite Hydrogel: Preparation and Characterization.

Polyvinyl alcohol/Chitosan hydrogel is often employed as a carrier because it is non-toxic, biodegradable, and has a three-dimensional network structure. Meanwhile, Magnesium-doped nano-hydroxyapatite(Mg-nHA) demonstrated high characterization to promote the osteogenic differentiation of bone marrow derived mesenchymal stem cell(BMSCs). Therefore, in order to develop a porous hydrogel scaffold for the application of bone tissue engineering, an appropriate-type Mg-nHA hydrogel scaffold was developed and evaluated. A composite hydrogel containing magnesium-doped nano-hydroxyapatite (Mg-nHA/PVA/CS) was developed using a magnetic stirring-ion exchange method and cyclic freeze-thaw method design, with polyvinyl alcohol and chitosan as the main components. Fourier transform infrared spectra (FTIR), electron energy dispersive spectroscopy (EDS), X-ray photoelectron spectrometer (XPS) and scanning electron microscopy (SEM) were employed to analyze the chemical structure, porosity, and elemental composition of each hydrogels. The equilibrium swelling degree, moisture content, pH change, potential for biomineralization, biocompatibility, the osteogenic potential and magnesium ion release rate of the composite hydrogel were also evaluated. SEM analysis revealed a well-defined 3D spatial structure of micropores in the synthesised hydrogel. FTIR analysis showed that doping nanoparticles had little effect on the hydrogel's structure and both the 5% Mg-nHA/PVA/CS and 10% Mg-nHA/PVA/CS groups promoted amide bond formation. EDS observation indicated that the new material exhibited favourable biomineralization ability, with optimal performance seen in the 5% Mg-nHA/PVA/CS group. The composite hydrogel not only displayed favourable water content, enhanced biocompatibility, and porosity (similar to human cancellous bone), but also maintained an equilibrium swelling degree and released magnesium ions that created an alkaline environment around it. Additionally, it facilitated the proliferation of bone marrow mesenchymal stem cells and their osteogenic differentiation. The Mg-nHA/PVA/CS hydrogel demonstrates significant potential for application in the field of bone repair, making it an excellent composite material for bone tissue engineering.