Scaffolds For Bone Regeneration Research Articles

Advances in the Application of Photothermal Composite Scaffolds for Osteosarcoma Ablation and Bone Regeneration.

Photothermal therapy is a promising approach to cancer treatment. The energy generated by the photothermal effect can effectively inhibit the growth of cancer cells without harming normal tissues, while the right amount of heat can also promote cell proliferation and accelerate tissue regeneration. Various nanomaterials have recently been used as photothermal agents (PTAs). The photothermal composite scaffolds can be obtained by introducing PTAs into bone tissue engineering (BTE) scaffolds, which produces a photothermal effect that can be used to ablate bone cancer with subsequent further use of the scaffold as a support to repair the bone defects created by ablation of osteosarcoma. Osteosarcoma is the most common among primary bone malignancies. However, a review of the efficacy of different types of photothermal composite scaffolds in osteosarcoma is lacking. This article first introduces the common PTAs, BTE materials, and preparation methods and then systematically summarizes the development of photothermal composite scaffolds. It would provide a useful reference for the combination of tumor therapy and tissue engineering in bone tumor-related diseases and complex diseases. It will also be valuable for advancing the clinical applications of photothermal composite scaffolds.

Dextran-Based Injectable Hydrogel Composites for Bone Regeneration.

Currently, bone infections caused by diseases or injuries are a major health issue. In addition, the conventional therapeutic approaches used to treat bone diseases or injuries present several drawbacks. In the area of tissue engineering, researchers have been developing new alternative therapeutic approaches, such as scaffolds, to promote the regeneration of injured tissues. Despite the advantages of these materials, most of them require an invasive surgical procedure. To overcome these problems, the main focus of this work was to develop scaffolds for bone regeneration, which can be applied using injectable hydrogels that circumvent the use of invasive procedures, while allowing for bone regeneration. Throughout this work, injectable hydrogels were developed based on a natural polymer, dextran, along with the use of two inorganic compounds, calcium β-triphosphate and nanohydroxyapatite, that aimed to reinforce the mechanical properties of the 3D mesh. The materials were chemically characterized considering the requirements for the intended application: the swelling capacity was evaluated, the degradation rate in a simulated physiological environment was assessed, and compression tests were performed. Furthermore, vancomycin was incorporated into the polymeric matrices to obtain scaffolds with antibacterial performance, and their drug release profile was assessed. The cytotoxic profile of the hydrogels was assessed by an MTS assay, using osteoblasts as model cells. The data obtained demonstrated that dextran-based hydrogels were successfully synthesized, with a drug release profile with an initial burst between 50 and 80% of the drug. The hydrogels possess fair biocompatibility. The swelling capacity showed that the stability of the samples and their degradation profile is compatible with the average time period required for bone regeneration (usually about one month) and have a favorable Young's modulus (200-300 kPa). The obtained hydrogels are well-suited for bone regeneration applications such as infections that occur during implantation or bone graft substitutes with antibiotics.

Advanced Synthetic Scaffolds Based on 1D Inorganic Micro-/Nanomaterials for Bone Regeneration.

Inorganic nanoparticulate biomaterials, such as calcium phosphate and bioglass particles, with chemical compositions similar to that of the inorganic component of natural bone, and hence having excellent biocompatibility and bioactivity, are widely used for the fabrication of synthetic bone graft substitutes. Growing evidence suggests that structurally anisotropic, or 1D inorganic micro-/nanobiomaterials are superior to inorganic nanoparticulate biomaterials in the context of mechanical reinforcement and construction of self-supporting 3D network structures. Therefore, in the past decades, efforts have been devoted to developing advanced synthetic scaffolds for bone regeneration using 1D micro-/nanobiomaterials as building blocks. These scaffolds feature extraordinary physical and biological properties, such as enhanced mechanical properties, super elasticity, multiscale hierarchical architecture, extracellular matrix-like fibrous microstructure, and desirable biocompatibility and bioactivity, etc. In this review, an overview of recent progress in the development of advanced scaffolds for bone regeneration is provided based on 1D inorganic micro-/nanobiomaterials with a focus on their structural design, mechanical properties, and bioactivity. The promising perspectives for future research directions are also highlighted.

Sustainable natural biopolymers for biomedical applications

Biopolymers are materials specifically engineered to interact with biological systems. They can be derived from either natural or synthetic sources, depending on the biological resources used or the manufacturing process employed. Over recent decades, these materials have gained significant popularity within the medical field due to their remarkable attributes such as biodegradability, bioactivity, and compatibility with human tissue. One notable application is their use as scaffolds for bone regeneration. Biopolymers, being renewable biomaterials, provide opportunities for continuous manufacturing and technological progress across various industries. These biomaterials have demonstrated great promise in medical sectors, including nerve regeneration and the production of surgical devices. Additionally, their versatility extends to non-biomedical applications, like food packaging. This paper aims to provide a comprehensive overview of different biopolymers, elucidating their properties, showcasing their latest applications, and delving into the state-of-the-art manufacturing technologies used in their production. Special emphasis is placed on their suitability as bone tissue repair and regeneration scaffolds, owing to their unique properties, which render them an ideal choice for this specific application.

Self-promoted electroactive biomimetic mineralized scaffolds for bacteria-infected bone regeneration

Infected bone defects are a major challenge in orthopedic treatment. Native bone tissue possesses an endogenous electroactive interface that induces stem cell differentiation and inhibits bacterial adhesion and activity. However, traditional bone substitutes have difficulty in reconstructing the electrical environment of bone. In this study, we develop a self-promoted electroactive mineralized scaffold (sp-EMS) that generates weak currents via spontaneous electrochemical reactions to activate voltage-gated Ca2+ channels, enhance adenosine triphosphate-induced actin remodeling, and ultimately achieve osteogenic differentiation of mesenchymal stem cells by activating the BMP2/Smad5 pathway. Furthermore, we show that the electroactive interface provided by the sp-EMS inhibits bacterial adhesion and activity via electrochemical products and concomitantly generated reactive oxygen species. We find that the osteogenic and antibacterial dual functions of the sp-EMS depend on its self-promoting electrical stimulation. We demonstrate that in vivo, the sp-EMS achieves complete or nearly complete in situ infected bone healing, from a rat calvarial defect model with single bacterial infection, to a rabbit open alveolar bone defect model and a beagle dog vertical bone defect model with the complex oral bacterial microenvironment. This translational study demonstrates that the electroactive bone graft presents a promising therapeutic platform for complex defect repair.

Mineralized Collagen/Polylactic Acid Composite Scaffolds for Load-Bearing Bone Regeneration in a Developmental Model.

Repairing load-bearing bone defects in children remains a big clinical challenge. Mineralized collagen (MC) can effectively simulate natural bone composition and hierarchical structure and has a good biocompatibility and bone conductivity. Polylactic acid (PLA) is regarded as a gold material because of its mechanical properties and degradability. In this study, we prepare MC/PLA composite scaffolds via in situ mineralization and freeze-drying. Cell, characterization, and animal experiments compare and evaluate the biomimetic properties and repair effects of the MC/PLA scaffolds. Phalloidin and DAPI staining results show that the MC/PLA scaffolds are not cytotoxic. CCK-8 and scratch experiments prove that the scaffolds are superior to MC and hydroxyapatite (HA)/PLA scaffolds in promoting cell proliferation and migration. The surface and interior of the MC/PLA scaffolds exhibit rich interconnected pore structures with a porosity of ≥70%. The XRD patterns are typical HA waveforms. X-ray, micro-CT, and H&E staining reveal that the defect boundary disappears, new bone tissue grows into MC/PLA scaffolds in a large area, and the scaffolds are degraded after six months of implantation. The MC/PLA composite scaffold has a pore structure and composition similar to cancellous bone, with a good biocompatibility and bone regeneration ability.

Amorphous iron-calcium phosphate-mediated biomineralized scaffolds for vascularized bone regeneration

Construction of organic–inorganic composites similar to natural bone in terms of structure and composition has attracted extensive attention. However, the clinical applications of these composites are limited due to the insufficient osteogenic and mechanical properties. In nature, the presence of amorphous iron-calcium phosphate (Fe-ACP) strengthens the mechanical properties of some biominerals, and our previous study has revealed its synthetic route and in vitro osteogenic properties. However, the potential role of Fe-ACP on biomineralization and constructing biomimetic scaffolds for bone regeneration has not been studied. Herein, a biomimetic scaffold with good osteogenic property was fabricated based on the mineralization of Fe-ACP, with the assistance of ice-templated freeze-casting. The in vitro study showed that the mineralized scaffolds possessed favorable biocompatibility and osteogenic property. Moreover, the scaffolds promoted cell chemotaxis and angiogenic property by upregulating the hypoxia inducible factor-1α (HIF-1α). In vivo experiment demonstrated potent early osteogenesis along with angiogenesis and ultimately promoted bone regeneration. Overall, the mineralized scaffold mediated by Fe-ACP precursors provide a unique platform to enhance bone tissue repair.

Review: Tailored microstructures in the system tricalcium phosphate-wollastonite-diopside for bone regeneration scaffolds

Tissue engineering constructs (TECs) combining resorbable scaffolds, cells and/or reactive agents are required for bone regeneration. New green ceramic processing methods allow fabricating layered and gradient porous structures. Dense bioceramics with sufficient strength that keep their microstructure when incorporating porosity are required. Bioceramics in the system tricalcium phosphate-wollastonite-diopside are reviewed. Tailoring of microstructures to improve their mechanical and biological behaviors and their potential for TECs are analyzed. First, a brief discussion on bone properties and requirements for biomaterials and the capabilities and limits of glasses and glass-ceramics formulated in the system is done. Then, the relationships between the microstructure and the mechanical behavior of single-phase and composite bioceramics are discussed. TCP-D-W microstructures with adequate mechanical performance support viability and osteogenic differentiation of human mesenchymal stem cells (hASCs) and their colonization by hASCs presents specific morphological features that make them adequate for TECs.

Biomimetic Structural Protein Based Magnetic Responsive Scaffold for Enhancing Bone Regeneration by Physical Stimulation on Intracellular Calcium Homeostasis.

The bone matrix has distinct architecture and biochemistry which present a barrier to synthesizing bone-mimetic regenerative scaffolds. To mimic the natural structures and components of bone, biomimetic structural decellularized extracellular matrix (ECM)/regenerated silk fibroin (RSF) scaffolds incorporated with magnetic nanoparticles (MNP) are prepared using a facile synthetic methodology. The ECM/RSF/MNP scaffold is a hierarchically organized and interconnected porous structure with silk fibroin twined on the collagen nanofibers. The scaffold demonstrates saturation magnetization due to the presence of MNP, along with good cytocompatibility. Moreover, the β-sheet crystalline domain of RSF and the chelated MNP could mimic the deposition of hydroxyapatite and enhance compressive modulus of the scaffold by ≈20%. The results indicate that an external static magnetic field (SMF) with a magnetic responsive scaffold effectively promotes cell migration, osteogenic differentiation, neogenesis of endotheliocytes in vitro, and new bone formation in a critical-size femur defect rat model. RNA sequencing reveals that the molecular mechanisms underlying this osteogenic effect involve calsequestrin-2-mediated Ca2+ release from the endoplasmic reticulum to activate Ca2+ /calmodulin/calmodulin-dependent kinase II signaling axis. Collectively, bionic magnetic scaffolds with SMF stimulation provide a potent strategy for bone regeneration through internal structural cues, biochemical composition, and external physical stimulation on intracellular Ca2+ homeostasis.

A biomimetic in situ mineralization ECM composite scaffold to promote endogenous bone regeneration

Bone tissue engineering scaffolds constructed from single-component organic materials have inherent limitations. Inspired by the hierarchical structure of physiological natural bone hard tissues, our research explores the construction of organic-inorganic composite scaffold for bone regeneration. In this study, we used a natural and readily obtainable extracellular matrix (ECM) material, i.e., decellularized small intestinal submucosa (SIS), to build the organic component of a phosphorylated hydroxyapatite nanocrystal-containing composite scaffold (nHA@SIS). Guided by polymer-induced liquid-precursor theory, we introduced a soluble inorganic mineralization solution to achieve an inorganic component of nHA@SIS. Using in situ mineralization, we successfully formed inorganic component within SIS and constructed nHA@SIS composite scaffold. We analyzed the physicochemical properties and the osteogenic role of nHA@SIS via a series of in vitro and in vivo studies. Compared with SIS scaffold, the nHA@SIS possessed suitable physicochemical properties, maintained the excellent cell activity of SIS and better guided reorganization of the cell skeleton, thereby achieving superior osteoconductivity and maintaining osteoinductivity at the protein and gene levels. Furthermore, the rat cranial defect area in the nHA@SIS scaffold group was mostly repaired after 12 weeks of implantation, with a larger amount of higher-density new bone tissue being visible at the edge and center than SIS and blank control group. This significantly improved in vivo osteogenic ability indicated the great potential of nHA@SIS for bone tissue engineering applications.

Physical stimuli-emitting scaffolds: The role of piezoelectricity in tissue regeneration.

The imbalance between life expectancy and quality of life is increasing due to the raising prevalence of chronic diseases. Musculoskeletal disorders and chronic wounds affect a growing percentage of people and demand more efficient tools for regenerative medicine. Scaffolds that can better mimic the natural physical stimuli that tissues receive under healthy conditions and during healing may significantly aid the regeneration process. Shape, mechanical properties, pore size and interconnectivity have already been demonstrated to be relevant scaffold features that can determine cell adhesion and differentiation. Much less attention has been paid to scaffolds that can deliver more dynamic physical stimuli, such as electrical signals. Recent developments in the precise measurement of electrical fields in vivo have revealed their key role in cell movement (galvanotaxis), growth, activation of secondary cascades, and differentiation to different lineages in a variety of tissues, not just neural. Piezoelectric scaffolds can mimic the natural bioelectric potentials and gradients in an autonomous way by generating the electric stimuli themselves when subjected to mechanical loads or, if the patient or the tissue lacks mobility, ultrasound irradiation. This review provides an analysis on endogenous bioelectrical signals, recent developments on piezoelectric scaffolds for bone, cartilage, tendon and nerve regeneration, and their main outcomes in vivo. Wound healing with piezoelectric dressings is addressed in the last section with relevant examples of performance in animal models. Results evidence that a fine adjustment of material composition and processing (electrospinning, corona poling, 3D printing, annealing) provides scaffolds that act as true emitters of electrical stimuli that activate endogenous signaling pathways for more efficient and long-term tissue repair.

A Study of the Properties of Scaffolds for Bone Regeneration Modified with Gel-like Coatings of Chitosan and Folic Acid.

The research has been conducted to obtain scaffolds for cancellous bone regeneration. Polylactide scaffolds were made by the phase inversion method with a freeze-extraction variant, including gelling polylactide in its non-solvent. Substitutes made of polylactide are hydrophobic, which limits cell adhesion. For this reason, the scaffolds were modified using chitosan and folic acid by forming gel-like coatings on the surface. The modification aimed to improve the material's surface properties and increase cell adhesion. Analyses of obtained scaffolds confirmed the effectiveness of performed changes. The presence of chitosan and folic acid was confirmed in the modified scaffolds, while all scaffolds retained high open porosity, which is essential for proper cell growth inside the scaffold and the free flow of nutrients. Hydrostatic weighing showed that the scaffolds have high mass absorbability, allowing them to be saturated with biological fluids. There were also cytotoxicity tests performed on 24 h extracts of the materials obtained, which indicated a lack of cytotoxic effect.

Influence of polylactide coating stereochemistry on mechanical and in vitro degradation properties of porous bioactive glass scaffolds for bone regeneration.

The mechanical properties of polylactide stereocomplexes (PLA SC) have been primarily studied through tensile testing, with inconsistent results, and the compressive properties of PLA SC compared to homocrystalline or amorphous PLA remain poorly understood. In this study, we coated porous bioactive glass 13-93 scaffolds with amorphous, homocrystalline, or stereocomplex PLA to investigate their mechanical and degradation properties before and after immersion in simulated body fluid. The glass scaffolds had interconnected pores and an average porosity of 76%. The PLA coatings, which were 10-100 μm thick and approximately 3% of the glass scaffold mass, covered the glass to a large extent. The compressive strength and toughness of all PLA-coated scaffolds were significantly higher than those of uncoated scaffolds, with approximately a fourfold increase before immersion and a twofold increase after immersion. The compressive strength and toughness of PLA SC-coated scaffolds were similar to those of scaffolds with homocrystalline PLA coating, and significantly higher than for scaffolds with amorphous PLA coating. All PLA coatings moderated the initial pH increase caused by the glass, which could benefit surrounding cells and bone tissue in vivo after implantation.

Hydroxyapatite 3D-printed scaffolds with Gyroid-Triply periodic minimal surface (TPMS) porous structure: Fabrication and an in vivo pilot study in sheep

Bone repair is a major challenge in regenerative medicine, e.g. for large defects. There is a need for bioactive, highly percolating bone substitutes favoring bone ingrowth and tissue healing. Here, a modern 3D printing approach (VAT photopolymerization) was exploited to fabricate hydroxyapatite (HA) scaffolds with a Gyroid-“Triply periodic minimal surface” (TPMS) porous structure (65% porosity, 90.5% HA densification) inspired from trabecular bone. Percolation and absorption capacities were analyzed in gaseous and liquid conditions. Mechanical properties relevant to guided bone regeneration in non-load bearing sites, as for maxillofacial contour reconstruction, were evidenced from 3-point bending tests and macrospherical indentation. Scaffolds were implanted in a clinically-relevant large animal model (sheep femur), over 6 months, enabling thorough analyses at short (4 weeks) and long (26 weeks) time points. In vivo performances were systematically compared to the bovine bone-derived Bio-OssⓇ standard. The local tissue response was examined thoroughly by semi-quantitative histopathology. Results demonstrated the absence of toxicity. Bone healing was assessed by bone dynamics analysis through epifluorescence using various fluorochromes and quantitative histomorphometry. Performant bone regeneration was evidenced with similar overall performances to the control, although the Gyroid biomaterial slightly outperformed Bio-OssⓇ at early healing time in terms of osteointegration and appositional mineralization. This work is considered a pilot study on the in vivo evaluation of TPMS-based 3D porous scaffolds in a large animal model, for an extended period of time, and in comparison to a clinical standard. Our results confirm the relevance of such scaffolds for bone regeneration in view of clinical practice. Statement of significanceBone repair, e.g. for large bone defects or patients with defective vascularization is still a major challenge.Highly percolating TPMS porous structures have recently emerged, but no in vivo data were reported on a large animal model of clinical relevance and comparing to an international standard.Here, we fabricated TPMS scaffolds of HA, determined their chemical, percolation and mechanical features, and ran an in-depth pilot study in the sheep with a systematic comparison to the Bio-OssⓇ reference.Our results clearly show the high bone-forming capability of such scaffolds, with outcomes even better than Bio-OssⓇ at short implantation time. This preclinical work provides quantitative data validating the relevance of such TMPS porous scaffolds for bone regeneration in view of clinical evaluation.

Comparative evaluation of personalized 3D-printed scaffold-driven double-barrel fibula flap for the reconstruction of segmented mandibular defects

The challenge of reconstructing large mandibular bone defects with double-barrel fibula flap (DBFF) is limited by the availability of autogenous bone, which remains a significant issue in maxillofacial surgery. This study aims to compare the novel scaffold-driven double-barrel approach with the DBFF method, and investigate the biomechanical behavior of 3D-printed porous titanium scaffolds with varying design parameters. The mandibular reconstruction model underwent FEA in two surgery phases, under two experimental loads and two design parameters. The experimental scaffold design was categorized into 9 groups based on the porous diameters (DP = 4 mm, 5 mm, 6 mm) and strut diameters (DS = 0.4 mm,0.5 mm,0.6 mm). The results illustrated that the stress in fibula, cortical and trabecular bones for all groups were within the ranges of normal physiological levels. The porous scaffolds effectively transmit mechanical stimulation, resulting in bone graft materials achieving up to 99.9% strain, with low stiffness to avoid stress shielding. The study revealed that the design parameters of Group B and Group F were more conducive to meeting biomechanical demands and promoting bone regeneration. This study presents scientifically proven and effective clinical solution for restoring both function and aesthetics of mandibular defects, while also providing valuable insights for future design criteria in bone regeneration scaffolds.

Interfacial characteristics of bioglass diffusion zone between scaffold-like hydroxyapatite/wollastonite coating and dense zirconia substrate

A thin bioglass (BG) layer or diffusion zone has been used to enhance interfacial bonding between co-sintered scaffold-like micro-porous hydroxyapatite (HA) coating over dense zirconia substrate. However, there still have problems such as low fracture toughness and low wear resistance. Therefore, while meeting the basic requirements of degradability and biocompatibility, how to improve the mechanical properties of the scaffold, endow it with osteoinductivity and build a low-density porous structure has become the focus of bone scaffold research. The tri-layer bio-ceramic composites, consisting of porous HA, BG diffusion zone, and strong zirconia substrate, are designed with superior loading-bearing capability to the scaffold. Surface handling properties of the scaffold-like HA coating are further enhanced by incorporating wollastonite. The gradient in material compositions across the coating and substrate interface is further minimized by adding HA into the zirconia substrate. Benefiting from these special design and the thin BG diffusion zone, the tri-layer bio-ceramics present excellent interfacial strength, which ensures good structural strength and machinability. The results could provide the feasible options for bio-ceramic scaffolds design and manufacturing process, thus advance the practical clinical applications of HA-based scaffolds for bone repair and regeneration.

MXene-Integrated Silk Fibroin-Based Self-Assembly-Driven 3D-Printed Theragenerative Scaffolds for Remotely Photothermal Anti-Osteosarcoma Ablation and Bone Regeneration

MXene-Integrated Silk Fibroin-Based Self-Assembly-Driven 3D-Printed Theragenerative Scaffolds for Remotely Photothermal Anti-Osteosarcoma Ablation and Bone Regeneration

Bioprinting of cell-laden protein-based hydrogels: From cartilage to bone tissue engineering

The fabrication of cell-laden protein-based hydrogels (PBHs) for bioprinting necessitates careful consideration of numerous factors to ensure optimal structure and functionality. Bioprinting techniques, such as single-cell, multi-cell, and cell aggregate bioprinting, are employed to encapsulate cells within PBHs bioink, enabling the creation of scaffolds for cartilage and bone regeneration. During the fabrication process, it is imperative to account for biophysical and biochemical factors that influence cell behavior and protein structure within the PBHs. Precise control of crosslinking methods, hydrogel rheological properties, and printing parameters is also crucial to achieve desired scaffold properties without compromising cell viability and protein integrity. This review primarily focuses on the influence of biophysical factors, including composition, microstructure, biodegradation, and crosslinking, as well as biochemical factors, including chemical structure, growth factors, and signaling molecules, on protein structure and cell behavior. Additionally, key considerations for bioprinting PBHs and their impact on the successful regeneration of tissues are discussed. Furthermore, the review highlights current advancements, existing challenges, and promising prospects in the development of cell-laden PBHs for bioprinting applications and the regeneration of bone and cartilage.

Development of Gelatin Methacryloyl/Sodium Alginate Interpenetrating Polymer Network Hydrogels for Bone Regeneration by Activating the Wnt/β-Catenin Signaling Pathway via Lithium Release.

Hydrogels have gained significant attention as biomaterials due to their remarkable properties resembling those of the extracellular matrix (ECM). In the present investigation, we successfully synthesized interpenetrating polymer network (IPN) hydrogels using gelatin methacryloyl (GelMA) and sodium alginate (SA), incorporating various concentrations of lithium chloride (LiCl; 0, 5, and 10 mM), aiming to develop a hydrogel scaffold for bone regeneration. Notably, the compressive modulus of the IPN hydrogels remained largely unaffected upon the inclusion of LiCl. However, the hydrogel with the high concentration of LiCl exhibited reduced fragmentation after compression testing. Intriguingly, we observed a significant improvement in cellular biocompatibility, primarily attributed to activation of the Wnt/β-catenin signaling pathway induced by LiCl. Subsequently, we evaluated the efficacy of the newly developed IPN-Li hydrogels in a rat cranial defect model and found that they substantially enhanced bone regeneration. Nevertheless, it is important to note that the introduction of high concentrations of LiCl did not significantly promote osteogenesis. This outcome can be attributed to the excessive release of Li+ ions into the extracellular matrix, hindering the desired effect. Overall, the IPN-Li hydrogel developed in this study holds great promise as a biodegradable material for bone regeneration applications.

Differentiation-inducing effect of osteoclast microgrooves for the purpose of three-dimensional design of regenerated bone.

In vivo bone remodeling is promoted by the balance between osteoclast and osteoblast activity. Conventional research on bone regeneration has mainly focused on increasing osteoblast activity, with limited studies on the effects of scaffold topography on cell differentiation. Here, we examined the effect of microgroove-patterned substrate with spacings ranging from 1 to 10 μm on the differentiation of rat bone marrow-derived osteoclast precursors. Tartrate-resistant acid phosphatase (TRAP) staining and relative gene expression quantification showed that osteoclast differentiation was enhanced in substrate with 1 µm microgroove spacing compared with that in the other groups. Additionally, the ratio of podosome maturation stages in substrate with 1 μm microgroove spacing exhibited a distinct pattern, which was characterized by an increase in the ratio of belts and rings and a decrease in that of clusters. However, myosin II abolished the effects of topography on osteoclast differentiation. Overall, these showed that the reduction of myosin II tension in the podosome core by an integrin vertical vector increased podosome stability and promoted osteoclast differentiation in substrates with 1 μm microgroove spacing, including that microgroove design plays an important role in scaffolds for bone regeneration. STATEMENT OF SIGNIFICANCE: Reduction of myosin II tension in the podosome core, facilitated by an integrin vertical vector, resulted in an enhanced osteoclast differentiation, concomitant with an increase in podosome stability within 1-μm-spaced microgrooves. These findings are anticipated to serve as valuable indicators for the regulation of osteoclast differentiation through the manipulation of biomaterial surface topography in tissue engineering. Furthermore, this study contributes to the lucidation of the underlying mechanisms governing cellular differentiation by providing insights into the impact of the microtopographical environment.