Functional Hydrogels in Bone Tissue Engineering: From Material Design to Translational Applications

Bone tissue engineering (BTE) provides an effective repair solution by implanting osteoblasts or stem cells into biocompatible and biodegradable scaffolds to promote bone regeneration. In recent years, the rapid development of 3D bioprinting has enabled its extensive application in fabricating BTE scaffolds. Based on three-dimensional computer models and specialized "bio-inks," this technology offers new pathways for customizing BTE scaffolds. This study reviews the current status and future prospects of scaffold materials for BTE in 3D bioprinting. This literature review collected recent studies on BTE and 3D bioprinting, analyzing the advantages and limitations of various scaffold materials for 3D printing, including bioceramics, metals, natural polymers, and synthetic polymers. Key characteristics like biocompatibility, mechanical properties, and degradation rates of these materials were systematically compared. The study highlights the diverse performances of materials used in BTE scaffolds. Bioceramics exhibit excellent biocompatibility but suffer from brittleness; metals offer high strength but may induce chronic inflammation; natural polymers are biocompatible yet have poor mechanical properties, while synthetic polymers offer strong tunability but may produce acidic by-products during degradation. Additionally, integrating 3D bioprinting with composite materials could enhance scaffold biocompatibility and mechanical properties, presenting viable solutions to current challenges. This review summarizes recent advances in 3D bioprinting for BTE scaffold applications, exploring the strengths and limitations of various materials and proposing composite material combinations to improve scaffold performance. By optimizing material selection and combinations, 3D bioprinting shows promise for creating customized scaffolds, offering a new technical route for clinical applications of BTE. This research provides a unique perspective and theoretical support for advancing 3D bioprinting technology in bone regeneration, outlining future directions for BTE materials and 3D bioprinting technology development.

Over the last few years, biopolymers have attracted great interest in tissue engineering and regenerative medicine due to the great diversity of their chemical, mechanical, and physical properties for the fabrication of 3D scaffolds. This review is devoted to recent advances in synthetic and natural polymeric 3D scaffolds for bone tissue engineering (BTE) and regenerative therapies. The review comprehensively discusses the implications of biological macromolecules, structure, and composition of polymeric scaffolds used in BTE. Various approaches to fabricating 3D BTE scaffolds are discussed, including solvent casting and particle leaching, freeze-drying, thermally induced phase separation, gas foaming, electrospinning, and sol–gel techniques. Rapid prototyping technologies such as stereolithography, fused deposition modeling, selective laser sintering, and 3D bioprinting are also covered. The immunomodulatory roles of polymeric scaffolds utilized for BTE applications are discussed. In addition, the features and challenges of 3D polymer scaffolds fabricated using advanced additive manufacturing technologies (rapid prototyping) are addressed and compared to conventional subtractive manufacturing techniques. Finally, the challenges of applying scaffold-based BTE treatments in practice are discussed in-depth.

Three-dimensional (3D) printing is a type of additive manufacturing that works by the application of material inks layer by layer using data from computer-aided design (CAD) to help to place the ink in a predefined place, thus producing a highly accurate product even with complex geometry. The goal in using 3D bioprinting is to develop a biological scaffold that resembles the desired tissue to be replaced, including the cells and the growth factors, in a specific spatial relationship. The developments in bone tissue engineering (BTE) and 3D bioprinting are revolutionizing osseous craniofacial reconstructive surgery. This chapter aims to describe 3D bioprinting of biomaterial and bioceramic scaffolds for bone tissue engineering and maxillofacial reconstructive surgery.KeywordsAdditive manufacturingLayer by layerBioprintingBiological scaffoldBioactive glassCalcium phosphateHydroxyapatiteMesenchymal stem cellsInduced pluripotent stem cellsExosomeBiomimeticsSelf-assembly

Background and Purpose: The growing demand for bone regeneration following severe injuries highlights the importance of scaffolds in bone tissue engineering (BTE). Injectable hydrogels have emerged as promising candidates because their properties closely mimic the native extracellular matrix (ECM). However, their limited mechanical strength and structural instability restrict their practical application. Approach: This review summarizes recent strategies for reinforcing in situ-forming injectable hydrogels to improve their mechanical performance for bone regeneration. Particular emphasis is placed on nanomaterial-based strategies, including the incorporation of nanoparticles and nanofibers, and their ability to enhance the physical properties of polymeric networks. Key Results: Evidence from recent studies demonstrates that reinforcing hydrogels with nano-scaled materials creates interconnected networks that improve load-bearing capacity, stability, and resistance to deformation. These reinforced systems retain the inherent advantages of injectable hydrogels-biocompatibility, biodegradability, permeability to oxygen and nutrients, and drug delivery capability-while addressing their mechanical shortcomings. Conclusion: Nanomaterial-based reinforcement offers a versatile approach to overcoming the limitations of injectable hydrogels in BTE. By providing improved structural integrity alongside biological functionality, these advanced systems broaden the potential of injectable hydrogels for clinical translation. Future work should focus on optimizing reinforcement strategies to balance mechanical enhancement with safety, manufacturability, and regulatory considerations.

Bacterial nanocellulose enhances osteogenic activity of Vat-based 3D-bioprinted silk fibroin hydrogels.

Nanofiber scaffolds have gained significant attention in the field of bone tissue engineering. Electrospinning, a straightforward and efficient technique for producing nanofibers, has been extensively researched. When used in bone tissue engineering scaffolds, electrospun nanofibers with suitable surface properties promote new bone tissue growth and enhance cell adhesion. Recent advancements in electrospinning technology have provided innovative approaches for scaffold fabrication in bone tissue engineering. This review comprehensively examines the utilization of electrospun nanofibers in bone tissue engineering scaffolds and evaluates the relevant literature. The review begins by presenting the fundamental principles and methodologies of electrospinning. It then discusses various materials used in the production of electrospun nanofiber scaffolds for bone tissue engineering, including natural and synthetic polymers, as well as certain inorganic materials. The challenges associated with these materials are also described. The review focuses on novel electrospinning techniques for scaffold construction in bone tissue engineering, such as multilayer nanofibers, multifluid electrospinning, and the integration of electrospinning with other methods. Recent advancements in electrospinning technology have enabled the fabrication of precisely aligned nanofiber scaffolds with nanoscale architectures. These innovative methods also facilitate the fabrication of biomimetic structures, wherein bioactive substances can be incorporated and released in a controlled manner for drug delivery purposes. Moreover, they address issues encountered with traditional electrospun nanofibers, such as mechanical characteristics and biocompatibility. Consequently, the development and implementation of novel electrospinning technologies have revolutionized scaffold fabrication for bone tissue engineering.

Scaffolds for Bone Tissue Engineering: State of the art and new perspectives

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.

Magnetic scaffolds for bone tissue engineering based on a poly(ε‐caprolactone) (PCL) matrix and iron oxide (Fe3O4) magnetic nanoparticles were designed and developed through a three‐dimensional (3D) fiber‐deposition technique. PCL/Fe3O4 scaffolds were characterized by a 90/10 w/w composition. Tensile and magnetic measurements were carried out, and nondestructive 3D imaging was performed through microcomputed tomography (Micro‐CT). Furthermore, confocal analysis was undertaken to investigate human mesenchymal stem cell adhesion and spreading on the PCL/Fe3O4 nanocomposite fibers. The results suggest that nanoparticles mechanically reinforced the PCL matrix; the elastic modulus and the maximum stress increased about 10 and 30%, respectively. However, the maximum strain decreased about 50%; this suggested an enhanced brittleness. Magnetic results evidenced a superparamagnetic behavior for these nanocomposite scaffolds. Micro‐CT suggested an almost uniform distribution of nanoparticles. Confocal analysis highlighted interesting results in terms of cell adhesion and spreading. All of these results show that a magnetic feature could be incorporated into a polymeric matrix that could be processed to manufacture scaffolds for advanced bone tissue engineering and, thus, provide new opportunity in terms of scaffold fixation and functionalization. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011

Hyaluronic acid (170 kDa)-based hydrogel was synthesized using acrylated hyaluronic acid (HA) and matrix metalloproteinase (MMP) sensitive HA-based hydrogels were then prepared by conjugation with two different peptides: cell adhesion peptides containing integrin-binding domains (Arg-Gly-Asp: RGD) and a cross-linker with MMP degradable peptides to mimic the remodeling characteristics of natural extracellular matrices by cell-derived MMPs. Mechanical properties of these hydrogels were evaluated with different weight percentages (2.5 and 3.5 wt %) by measuring elastic modulus, viscous modulus, and swelling ratio. Human mesenchymal stem cells (hMSCs) were then cultured in MMP-sensitive or insensitive HA-based hydrogels and/or immobilized cell adhesive RGD peptides in vitro. Actin staining and image analysis proved that cells cultured in the MMP-sensitive hydrogel with RGD peptides showed extensive cell spreading and sprouting. Gene expression analysis showed that bone specific genes such as alkaline phosphatase, osteocalcin, and osteopontin increased in MMP-sensitive hydrogels as biomolecules such as BMPs and cells were added in the gels. For in vivo calvarial defect regeneration, five different samples (MMP insensitive hydrogel, MMP sensitive hydrogel, MMP sensitive hydrogel with BMP-2, MMP sensitive hydrogel with hMSC, and MMP sensitive hydrogel with BMP-2 and hMSC) were prepared. After 4 weeks of implantation, the Masson-Trichrome staining and micro computed tomography scan results demonstrated that the MMP sensitive hydrogels with BMP-2 and hMSCs have the highest mature bone formation. The MMP sensitive HA-based hydrogel could become useful scaffolds in bone tissue engineering with improvements on tissue remodeling rates and regeneration activity.

Scaffolds used for bone tissue engineering need to have a variety of features to accommodate bone cells. The scaffold should mimic natural bone, it should have appropriate mechanical strength, support cell differentiation to the osteogenic lineage, and offer adequate porosity to allow vascularization and bone in-growth. In this work, we aim at developing a new process to fabricate such materials by creating a porous composite material made of silk fibroin and cellulose as a suitable scaffold of bone tissue engineering. Silk fibroin and cellulose are both dissolved together in N,N-dimethylacetamide/LiCl and molded to a porous structure using NaCl powder. The hydrogels are prepared by a sequential regeneration process: cellulose is solidified by water vapor treatment, while the remaining silk fibroin in the hydrogel is insolubilized by methanol, which leads to a cellulose framework structure embedded in a silk fibroin matrix. Finally, the hydrogels are soaked in water to dissolve the NaCl for making a porous structure. The cellulose composition results in improving the mechanical properties for the hydrogels in comparison to the silk fibroin control material. The pore size and porosity are estimated at around 350 µm and 70%, respectively. The hydrogels support the differentiation of MC3T3 cells to osteoblasts and are expected to be a good scaffold for bone tissue engineering.

Composite bioink incorporating cell-laden liver decellularized extracellular matrix for bioprinting of scaffolds for bone tissue engineering

Electrospinning is a promising and versatile technique that is used to fabricate polymeric nanofibrous scaffolds for bone tissue engineering. Ideal scaffolds should be biocompatible and bioactive with appropriate surface chemistry, good mechanical properties and should mimic the natural extracellular matrix (ECM) of bone. Selection of the most appropriate material to produce a scaffold is an important step towards the construction of a tissue engineered product. Bone tissue engineering is an interdisciplinary field, where the principles of engineering are applied on bone-related biochemical reactions. Scaffolds, cells, growth factors, and their interrelation in microenvironment are the major concerns in bone tissue engineering. This review covers the latest development of biomimetic electrospun polymeric biomaterials for bone tissue engineering. It includes the brief details to bone tissue engineering along with bone structure and ideal bone scaffolds requirements. Details about various engineered materials and methodologies used for bone scaffolds development were discussed. Description of electrospinning technique and its parameters relating their fabrication, advantages, and applications in bone tissue engineering were also presented. The use of synthetic and natural polymers based electrospun nanofibrous scaffolds for bone tissue engineering and their biomineralization processes were discussed and reviewed comprehensively. Finally, we give conclusion along with perspectives and challenges of biomimetic scaffolds for bone tissue engineering based on electrospun nanofibers.

In bone tissue engineering, a suitable scaffold is the key. Due to their similar composition to bone tissue, special structure, good mechanical properties, and osteogenic properties, acellular fish scale scaffolds are potential scaffolds for bone tissue engineering. At present, the fish scale decellularization scheme mostly uses a combination of sodium dodecyl sulfate and ethylenediamine tetraacetic acid (EDTA), but this method has problems. We optimized this method using a combined method of Triton X-100, EDTA, and nuclease. In this study, the optimal scheme was screened with respect to the decellularization effect, extracellular matrix composition and structure retention, mechanical properties, cell biocompatibility, and osteogenic differentiation ability. The results showed that the optimal scheme was as follows: the native fish scales were incubated in 0.1% EDTA for 24 h, and then the cellular components were removed with 1% Triton X-100 for 4 days, followed by nuclease digestion for 24 h. On that basis, we proposed a novel and more suitable fish scale decellularization scheme, and the acellular fish scale scaffold prepared by this decellularization scheme may have great potential in bone tissue engineering.

Currently available bone grafts present certain limitations such as donor-site morbidity for autografts and risk of disease transmission for allografts. These limitations suggest a need for alternative strategies. The use of polymer/ceramic composites as scaffolds for trabecular bone tissue engineering capitalizes on the benefits of both materials. Polymers are easily formed and shaped and add structural rigidity while calcium phosphates impart osteoconductivity and osteointegration. We have developed a biodegradable, microsphere-based scaffold for bone tissue engineering based on poly(lactide-co-glycolide)/calcium phosphate composite microspheres in which a nanocrystalline hydroxyapatite is synthesized within the forming microspheres.