3D-printed Titanium Alloy Research Articles

Fabrication of a composite 3D-printed titanium alloy combined with controlled in situ drug release to prevent osteosarcoma recurrence

Osteosarcoma is a malignant bone tumor occurring in adolescents. Surgery combined with adjuvant or neoadjuvant chemotherapy is the standard treatment. However, systemic chemotherapy is associated with serious side effects and a high risk of postoperative tumor recurrence, leading to a high amputation rate and mortality in cancer patients. Implant materials that can simultaneously repair large bone defects and prevent osteosarcoma recurrence are in urgent need. Herein, an intelligent system comprising 3D-printed titanium scaffold (TS) and pH-responsive PEGylated paclitaxel prodrugs was fabricated for bone defect reconstruction and recurrence prevention following osteosarcoma surgery. The drug-loaded implants exhibited excellent stability and biocompatibility for supporting the activity of bone stem cells under normal body fluid conditions and the rapid release of drugs in response to faintly acidic environments. An in vitro study demonstrated that five human osteosarcoma cell lines could be efficiently eradicated by paclitaxel released in an acidic microenvironment. Using mice models, we demonstrated that the drug-loaded TS can enable a pH-responsive treatment of postoperative tumors and effectively prevent osteosarcoma recurrence. Therefore, local implantation of this composite scaffold may be a promising topical therapeutic method to prevent osteosarcoma recurrence.

3D-Printed Porous Tantalum Scaffold Improves Muscle Attachment via Integrin-β1-Activated AKT/MAPK Signaling Pathway.

3D-printed porous titanium (Ti) alloy scaffolds have been reported for facilitating muscle attachment in our previous study. However, the anti-avulsion ability needs to be improved. In this study, we used 3D-printed porous tantalum (Ta) scaffolds to improve muscle attachment. The differences in chemical and physical characteristics and muscle adhesion between the two scaffolds were tested and compared in the gene and protein level both in vitro and in vivo. The possible molecular mechanism was analyzed and further proved. The results showed that compared with the porous Ti alloy, porous Ta had better cell proliferation, differentiation, migration, and adhesion via the integrin-β1 (Itgb1)-activated AKT/MAPK signaling pathway in L6 rat myoblasts. When artificially down-regulated the expression of Itgb1, cell adhesion and myogenesis differentiation were affected and the phosphorylation of the AKT/MAPK signaling pathway was suppressed. In rat intramuscular implantation, porous Ta had a significantly higher muscle ingrowth rate (85.63% ± 4.97 vs 65.98% ± 4.52, p < 0.01) and larger avulsion force (0.972 vs 0.823 N/mm2, p < 0.05) than the porous Ti alloy. These findings demonstrate that the 3D-printed porous Ta scaffold is beneficial for further clinical application of muscle attachment.

3D Printing Technology for Chest Wall Reconstruction With a Sternum-Ribs-Cartilage Titanium Implant: From Ideation to Creation.

We describe the use of 3-dimensional (3D) printing technology to plan and reconstruct the sternum, adjoining cartilages, and ribs with a custom-made, anatomically designed, 3D-printed titanium implant for an isolated sternal metastasis complicated with a pathological fracture. We imported submillimeter slice computed tomography scan data into Mimics Medical 20.0 software and by manual bone threshold segmentation created a 3D virtual model of the patient's chest wall and tumor. For all-around tumor-free margins, we grew the tumor by 2 cm. The replacement implant was designed in 3D using the anatomical features of the sternum, cartilages, and ribs and manufactured using TiMG 1 powder fusion technology. Physiotherapy was provided prior to and following surgery, and the impact of reconstruction on pulmonary functions was assessed. At surgery, the precise resection, clear margins, and a secure fit were achieved. At follow-up, there was no dislocation, paradoxical movement, change in performance status, or dyspnea. There was a decrease in forced expiratory volume in 1 s (FEV1) from 105% prior to surgery to 82% following surgery and in forced vital capacity (FVC) from 108% to 75%, with no difference in the FEV1/FVC ratio, suggesting a restrictive pattern of impairment. With 3D printing technology, reconstructing a large anterior chest wall defect with a custom-made, anatomical, 3D-printed titanium alloy implant is feasible and safe, and it preserves the shape, structure, and function of the chest wall, albeit with a restrictive pattern of pulmonary function, which can be addressed with physiotherapy.

Construction and osteogenic effects of 3D-printed porous titanium alloy loaded with VEGF/BMP-2 shell-core microspheres in a sustained-release system.

The repair and reconstruction of bone defects remain a challenge in orthopedics. The present study offers a solution to this problem by developing a vascular endothelial growth factor (VEGF)/bone morphogenetic protein 2 (BMP-2) shell-core microspheres loaded on 3D-printed porous titanium alloy via gelatin coating to prepare a titanium-alloy microsphere scaffold release system. The composite scaffold was characterized via scanning electron microscope (SEM) and energy disperse spectroscopy (EDS), and the effect of the composite scaffold on the adhesion, proliferation, and differentiation of osteoblasts were determined in vitro. Furthermore, a rabbit femoral defect model was established to verify the effect of the composite scaffold on osteogenesis and bone formation in vivo. The results demonstrated that the composite scaffold could release VEGF and BMP-2 sequentially. Meanwhile, the composite scaffold significantly promoted osteoblast adhesion, proliferation, and differentiation (p < 0.05) compared to pure titanium alloy scaffolds in vitro. Furthermore, the composite scaffold can exhibit significant osteogenic differentiation (p < 0.05) than gelatin-coated titanium alloy scaffolds. The in vivo X-rays demonstrated that the implanted scaffolds were in a good position, without inflammation and infection. Micro-CT and quantitative results of new bone growth illustrated that the amount of new bone in the composite scaffold is significantly higher than that of the gelatin-coated and pure titanium alloy scaffolds (p < 0.05). Similarly, the fluorescence labeling and V-G staining of hard tissue sections indicated that the bone integration capacity of the composite scaffold was significantly higher than the other two groups (p < 0.05). This research suggests that VEGF/BMP-2 shell-core microspheres loaded on 3D-printed titanium alloy porous scaffold through gelatin hydrogel coating achieved the sequential release of VEGF and BMP-2. Most importantly, the in vitro and in vivo study findings have proven that the system could effectively promote osteogenic differentiation and osseointegration.

Length scale parameters to estimate fatigue lifetime of 3D-printed titanium alloy Ti6Al4V containing notches in the as-manufactured condition

The accuracy of the Theory of Critical Distances, Gradient Elasticity and the Averaged Strain Energy Density criterion in estimating fatigue lifetime of notched additively manufactured Ti6Al4V is assessed against numerous experimental results generated under load ratios equal to −1 and 0.1. The 3D-printed Ti-alloy under investigation was tested by keeping the notches in the as-manufactured condition. The common feature of the considered design approaches is that they all make use of a material length scale. The validation exercise based on the generated experimental results demonstrates that the length scale concept can be extended successfully also to the fatigue assessment of notched 3D-printed metallic components.

An in vitro trial on the effect of arch form on connector size requirements in long span anterior zirconia fixed dental prostheses.

To test the fracture resistance of maxillary canine to canine fixed partial denture with four missing incisors, with increasing anterior-cantilevers of the pontics and varying connector sizes. Two 3D-printed titanium alloy (Ti6Al4V) models mimicking a maxillary canine to canine fixed partial denture (FPD) with four pontics replacing the incisors were used as master models. Zirconia FPDs were digitally designed and milled with two different connector sizes (9 and 12 mm2 ) each with three different anterior cantilevers (7, 10, and 13mm) accounting for 6 test groups. Seven samples were milled for each group generating a total of 42 samples. The zirconia FPDs were cemented on the titanium model using resin modified glass ionomer cement and the model fixated to a variable angle vice. A sinusoidal cyclic waveform load from 50 to 280N was applied using a universal testing machine at a frequency of 30 cycles per second and a total of 5 million cycles. The results of Fisher's exact tests showed that the difference in the proportion of fractured versus nonfractured fixed partial dentures was not statistically significant when comparing the 9 with the 12 mm2 connector size (p = 1.00), as well as when comparing the six test groups (p = 0.2338); on the other hand, it proved to be statistically significant when comparing the 7mm cantilever with the 10 and 13mm cantilevers combined (p = 0.0407) indicating that a 7mm anterior spread of the pontics showed a significantly greater proportion of fixed partial dentures that fractured. Fracture susceptibility was not a function of cantilever length in this testing configuration for anterior FPDs. Retainer crown thickness seems to be a more important parameter than connector size thickness. Based on the results, a smaller connector size (9 mm2 ) can be used to improve the esthetics of pontics in long span anterior FPDs.

In vivo evaluation of osseointegration ability of sintered bionic trabecular porous titanium alloy as artificial hip prosthesis.

Hydroxyapatite (HA) coatings have been widely used for improving the bone-implant interface (BII) bonding of the artificial joint prostheses. However, the incidence of prosthetic revisions due to aseptic loosening remains high. Porous materials, including three-dimensional (3D) printing, can reduce the elastic modulus and improve osseointegration at the BII. In our previous study, we identified a porous material with a sintered bionic trabecular structure with in vitro and in vivo bio-safety as well as in vivo mechanical safety. This study aimed to compare the difference in osseointegration ability of the different porous materials and HA-coated titanium alloy in the BII. We fabricated sintered bionic trabecular porous titanium acetabular cups, 3D-printed porous titanium acetabular cups, and HA-coated titanium alloy acetabular cups for producing a hip prosthesis suitable for beagle dogs. Subsequently, the imaging and histomorphological analysis of the three materials under mechanical loading in animals was performed (at months 1, 3, and 6). The results suggested that both sintered bionic porous titanium alloy and 3D-printed titanium alloy exhibited superior performances in promoting osseointegration at the BII than the HA-coated titanium alloy. In particular, the sintered bionic porous titanium alloy exhibited a favorable bone ingrowth performance at an early stage (month 1). A comparison of the two porous titanium alloys suggested that the sintered bionic porous titanium alloys exhibit superior bone in growth properties and osseointegration ability. Overall, our findings provide an experimental basis for the clinical application of sintered bionic trabecular porous titanium alloys.

In Vitro and In Vivo Analysis of the Effects of 3D-Printed Porous Titanium Alloy Scaffold Structure on Osteogenic Activity.

The effect of titanium scaffold geometry on the bone regeneration ability of the scaffold remains unclear. Here, selective laser melting as a 3D printing technology was used to create porous titanium alloy scaffolds with two unit structures: a hollow hexagonal prism (group A) and a hollow triangular prism (group B). The structures and morphologies of the scaffolds were characterized before mechanical properties were simulated. Cell adhesion behaviors, osteoblast activity and proliferation, and alkaline phosphatase (ALP) activity were evaluated, in addition to in vivo testing in an animal model. The results showed that the two scaffolds made of Ti6Al4V had compression moduli similar to that of human cortical bone (116.91 ± 0.01 and 174.29 ± 2.21 MPa vs. 89–164 MPa). The two scaffolds were nontoxic to cells and had good biocompatibility, while group A scaffolds facilitated cell adhesion. The number of cells increased gradually in culture. The ALP activity of cells on group A scaffolds demonstrated higher osteogenic ability than that of group B scaffolds. The in vivo tests showed that all scaffolds retained their shape well after implantation, and no obvious inflammatory reaction or infection in surrounding tissues was found. Based on fluorescence staining, mature new bone formation was found at week 12. Group A scaffolds showed better bone integration ability compared with group B scaffolds. The percentage of new bone area in group A (7.5%) was higher than that in group B (6.5%). This research suggests that the hollow hexagonal prism structure of porous scaffolds can promote osteogenic differentiation and osseointegration better than the triangular prism structure.

In vivo study of dual functionalized mussel-derived bioactive peptides promoting 3D-printed porous Ti6Al4V scaffolds for repair of rabbit femoral defects.

The 3D printed porous titanium alloy scaffolds are beneficial to enhance angiogenesis, osteoblast adhesion, and promote osseointegration. However, titanium alloys are biologically inert, which makes the bond between the implant and bone tissue weak and prone to loosening. Inspired by the natural biological marine mussels, we designed four-claw-shaped mussel-derived bioactive peptides for the decoration of porous titanium alloy scaffolds: adhesion peptide-DOPA, anchoring peptide-RGD and osteogenic-inducing peptide-BMP-2. And the bifunctionalization of 3D-printed porous titanium alloy scaffolds was evaluated in vivo in a rabbit model of bone defect with excellent promotion of osseointegration and mechanical stability. Our results show that the in vivo osseointegration ability of the modified 3D printed porous titanium alloy test piece is significantly improved, and the bifunctional polypeptide coating group E has the strongest osseointegration ability. In conclusion, our experimental design partially solves the problems of stress shielding effect and biological inertness, and provides a convenient and feasible method for the clinical application of titanium alloy implants in biomedical implant materials.

Material parameters identification of 3D printed titanium alloy prosthesis stem based on response surface method

3D printed Titanium alloy is widely used as a material of artificial joints and its mechanical properties is a key factor for improving operation results. Because the elastic modulus of the 3 D printed titanium alloy specimen was related to the size of the metal blank. It is very difficult to identify mechanical parameters by traditional mechanics experiments. In this paper, according to the inverse analysis principle of the parameter estimation, a response surface methodology (RSM) was proposed to identify the mechanical parameters, based on finite element inverse analysis. The finite element models of femoral prosthesis stem were established in line with compression experiments. The material parameters were combined by central composite design (CCD), and the response surface (RS) models were constructed using a quadratic polynomial with cross terms and optimized using a genetic algorithm (GA). Finally, the best mechanical parameter combination of the femoral prosthesis was calculated. The calculated elastic modulus and Poisson's ratio of a 3 D printed titanium alloy femoral prosthesis stem were 109.07 GPa and 0.29, respectively, with the elastic modulus error being very small. The proposed method is effective and can be extended for the identification of mechanical parameters in other 3 D printed models.

Quercetin-Coating Promotes Osteogenic Differentiation, Osseointegration and Anti-Inflammatory Properties of Nano-Topographic Modificated 3D-Printed Ti6Al4V Implant.

The capabilities of osseointegration and anti-inflammatory properties are of equal significance to the bio-inert titanium implant surface. Quercetin has proved its capacities of activating anti-inflammation through macrophage modulation and promoting osteogenic differentiation. Herein, we fabricated quercetin-coating on nano-topographic modificated 3D-printed Ti6Al4V implant surface. Subsequently the biological cells responses in vitro, anti-inflammatory and osseointegration performance in vivo were evaluated. In vitro studies indicated that quercetin-coating can enhance the adhesion and osteogenic differentiation of rBMSCs, while modulating the polarization of macrophages from M1 to M2 phase and improving the anti-inflammatory and vascular gene expression. Moreover, quercetin-loaded implants reduced the level of peri-implant inflammation and ameliorated new bone formation and rapid osseoinegration in vivo. Quercetin-coating might provide a feasible and favorable scheme for endowing 3D-printed titanium alloy implant surface with enhanced the rapid osseointegration and anti-inflammatory properties.

Corrosion Resistance of 3D-Printed Titanium Alloy Ti64-ELI Parts for Dental Application.

Corrosion in the human body can cause materials to change structurally and release undesirable products that may bring about complications such as toxicity and inflammation. These may jeopardize the mechanical stability of prostheses. The purpose of this study was to evaluate the effect of solutions (Ringer's and table salt [NaCl]) and immersion periods on 3D-printed titanium alloy Ti64-ELI samples and the changes in mechanical properties before and after corrosion testing. The microstructure of prepared samples was analyzed, and the formation of α- and β-phases was studied. During testing, the β-phases showed up as white, and the α-phases presented as dark. In both, corrosion by pitting was observed after corrosion analysis. The results show that, by comparing NaCl and Ringer's, the Ecorr of the solutions increased by 0.8 V and the Icorr decreased by an order of magnitude. It was observed that the weight loss in the solutions will lead to dental implant instability and will cause failure.

Repair of distal fibular and lateral malleolus defects with individualized 3D-printed titanium alloy prosthesis: The first case report from China.

Introduction and importanceThis case report describes the reconstruction of the traumatic distal fibular and lateral malleolus defects with a novel method of using individualized 3D printed titanium prosthesis for the first time.Case presentationA 63-year-old male farmer was hospitalized in emergency because of open injury and distal fibular and lateral malleolus defects in the left leg caused by a car accident. 3 months after debridement and latissimus dorsi muscle flap transplantation and skin graft operation, the patient re-hospitalized because of distal fibular and lateral malleolus defect and local pain. We examined the bilateral ankle joint with three-dimensional CT, obtained data about the missing left distal fibular and lateral malleolus through the mirror principle. The corresponding titanium alloy prosthesis then was designed and printed using a 3D metal printer. The patient had no obvious contraindication for surgery, so the prosthesis was surgically implanted. The patient was followed up for 2 years. There was no discomfort at the surgical site. The function of the operated ankle was satisfied by the patient, the AOFAS (American Orthopaedic Foot & Ankle Society) score was 85 (Kitaoka et al., 1994 [1]).Clinical discussionIndividualized 3D printed titanium alloy prosthesis consistent with the anatomical structure of lost distal fibula and lateral malleolus. The proximal end of the prosthesis was designed with four nail holes to install screws to fix the fibula together with it. The lower tibiofibular and talofibular joint surfaces of the prosthesis were designed smoothly. In order to improve the stability of the lower tibiofibular joint, anchors were placed at the attachment of the anterior and posterior tibiofibular ligaments to reconstruct these ligaments.ConclusionThe structure and function of the reconstructed distal fibular and the lateral malleous were close to normal. Individualized 3D printed prosthesis might have considerable advantages over traditional treatment methods. The individualized 3D printed titanium alloy prosthesis provides a new method for the repair and reconstruction of similar bone defects. The use of 3D printed prosthesis for surgical repair needs to be further examined in the future through long-term follow-up studies and in more cases.

In vitro behaviour of human gingival fibroblasts cultured on 3D-printed titanium alloy with hydrogenated TiO2 nanotubes

Selective laser melting (SLM), as one of the most common 3D-printed technologies, can form personalized implants, which after further surface modification can obtain excellent osseointegration. To study the surface properties of SLM titanium alloy (Ti6Al4V) with hydrogenated titanium dioxide (TiO2)nanotubes (TNTs) and its influence on the biological behaviour of human gingival fibroblasts (HGFs), we used SLM to prepare 3D-printed titanium alloy samples (3D-Ti), which were electrochemically anodizing to fabricate 3D-TNTs and then further hydrogenated at high temperature to obtain 3D-H2-TNTs. Polished cast titanium alloy (MP-Ti) was used as the control group. The surface morphology, hydrophilicity and roughness of MP-Ti, 3D-Ti, 3D-TNTs and 3D-H2-TNTs were measured and analysed by scanning electron microscopy (SEM), contact angle metre, surface roughness measuring instrument and atomic force microscope, respectively. HGFs were cultured on the four groups of samples, and the cell morphology was observed by SEM. Fluorescence staining (DAPI) was used to observe the number of adhered cell nuclei, while a cell counting kit (CCK-8) was used to detect the early adhesion and proliferation of HGFs. Fluorescence quantitative real time polymerase chain reaction (RT–qPCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the expression of adhesion-related genes and fibronectin (FN), respectively. The results of this in vitro comparison study indicated that electrochemical anodic oxidation and high-temperature hydrogenation can form a superhydrophilic micro-nano composite morphology on the surface of SLM titanium alloy, which can promote both the early adhesion and proliferation of human gingival fibroblasts and improve the expression of cell adhesion-related genes and fibronectin.Graphical abstract

3D-Printed Bionic Titanium Alloy Artificial Lamina Prevents Epidural Adhesion and Restores the Stability After Laminectomy in Pigs.

Laminectomy can cause the dura mater to adhere to the surrounding scar tissue, leading to soft spinal stenosis after surgery. Although artificial laminae are considered ideal substitutes, they present challenges such as insecure fixation and insufficient bionics. In this study, we fabricated a bionic titanium alloy artificial lamina using three-dimensional (3D)-printing technology and evaluated its adhesion prevention and stability after laminectomy in pigs. An in vitro biomechanical pull-out resistance test indicated that the pull-out strength of the artificial lamina was close to that of a single pedicle screw and was significantly higher than that of a cortical screw. In vivo animal implantation results indicated precise laminectomy and artificial lamina implantation, as well as a safe operation process with the assistance of guide plates. X-ray and computed tomography results indicated the well fixation of bionic titanium alloy artificial lamina and screws 10 weeks after laminectomy. The artificial lamina was not loosened after being removed from pigs (postoperative week 12), exhibiting good stability. Additionally, no adhesion was observed in the artificial lamina group, whereas a large amount of scar tissue in the spinal canal covered the dural surface in the control group. Thus, 3D-printed bionic titanium alloy artificial lamina can prevent epidural adhesion after laminectomy, while restoring the structural stability of the posterior complex, suggesting the potential of lamina substitutes for adhesion prevention after laminectomy.

Advanced Surface Modification for 3D-Printed Titanium Alloy Implant Interface Functionalization.

With the development of three-dimensional (3D) printed technology, 3D printed alloy implants, especially titanium alloy, play a critical role in biomedical fields such as orthopedics and dentistry. However, untreated titanium alloy implants always possess a bioinert surface that prevents the interface osseointegration, which is necessary to perform surface modification to enhance its biological functions. In this article, we discuss the principles and processes of chemical, physical, and biological surface modification technologies on 3D printed titanium alloy implants in detail. Furthermore, the challenges on antibacterial, osteogenesis, and mechanical properties of 3D-printed titanium alloy implants by surface modification are summarized. Future research studies, including the combination of multiple modification technologies or the coordination of the structure and composition of the composite coating are also present. This review provides leading-edge functionalization strategies of the 3D printed titanium alloy implants.

Comparison of 3D-printed Titanium Alloy and Polyether Ether Ketone Prosthetic Beaks for an Injured Red-crowned Crane (Grus japonensis).

The red-crowned crane (Grus japonensis) is a critically endangered species. Three-dimensional-printed prosthetic beaks made of titanium alloy and polyether ether ketone (PEEK) were used to repair the beak of a red-crowned crane that had a complete fracture of the anterior maxillary bone and rhinotheca. The physical properties and stability of the prostheses and changes in the crane's behaviors after application of either beak were evaluated. The titanium alloy and PEEK prosthetic beaks weighed 30.81 g and 5.9 g, respectively. Scanning electron microscopy showed differences in microstructure between the 2 materials and the true beak; the true beak was softer than both materials from which the prostheses were made. The titanium beak frequently detached, and the residual natural beak showed significant cuticle softening with this prosthetic beak. The titanium beak detached within an approximately 3-month period after placement, whereas the PEEK prosthetic beak has remained secure for 2 years. Moreover, the crane's foraging times (P < .01) and grooming times (P < .05) with the titanium alloy false beak were lower than the normal, red-crowned crane. With the PEEK beak, no detachment or cuticle softening occurred, and foraging and grooming behaviors were evaluated by the investigators as natural (P > .05). Based on the results of this clinical case, the PEEK prosthetic beak was found to be superior to the titanium alloy prosthetic beak in color, weight, firmness, and postoperative effects.

Front Cover: Tailoring Additively Manufactured Titanium Implants for Short‐Time Pediatric Implantations with Enhanced Bactericidal Activity (ChemMedChem 2/2022)

The Front Cover illustrates fabricated 3D-printed titanium alloy model implants modified by electrochemical anodization to generate vertically aligned titania nanotubes (TNTs) on the surface with specific diameters (∼100 nm) aiming to reduce bone cell attachment and proliferation. To achieve enhanced antibacterial performance, the nanotubes were loaded with gallium nitrate as antibacterial agent. The physicochemical characteristics of these implants confirmed their ability to reduce osteoblast cell MG-63 cells prefiltration and overgrowth and strong antibacterial activity against S. aureus and P. aeruginosa with supressed bacterial attachment and high rates of bacterial death. The engineering of theses temporary Ti implants with enhanced bactericidal properties with reduced bone cell attachment is proposed as new strategy for designing of short-term implants in paediatrics. Cover design by Raymediasa. More information can be found in the Full Paper by Shaheer Maher, Dusan Losic et al.

Titanium alloy composited with dual-cytokine releasing polysaccharide hydrogel to enhance osseointegration via osteogenic and macrophage polarization signaling pathways.

Titanium alloy has been widely used in orthopedic surgeries as bone defect filling. However, the regeneration of high-quality new bones is limited due to the pro-inflammatory microenvironment around implants, resulting in a high occurrence rate of implant loosening or failure in osteological therapy. In this study, extracellular matrix-mimetic polysaccharide hydrogel co-delivering BMP-2 and interleukin (IL)-4 was composited with 3D printed titanium alloy to promote the osseointegration and regulate macrophage response to create a pro-healing microenvironment in bone defect. Notably, it is discovered from the bioinformatics data that IL-4 and BMP-2 could affect each other through multiple signal pathways to achieve a synergistic effect toward osteogenesis. The composite scaffold significantly promoted the osteoblast differentiation and proliferation of human bone marrow mesenchyme stem cells (hBMSCs). The repair of large-scale femur defect in rat indicated that the dual-cytokine-delivered composite scaffold could manipulate a lower inflammatory level in situ by polarizing macrophages to M2 phenotype, resulting in superior efficacy of mature new bone regeneration over the treatment of native titanium alloy or that with an individual cytokine. Collectively, this work highlights the importance of M2-type macrophages-enriched immune-environment in bone healing. The biomimetic hydrogel–metal implant composite is a versatile and advanced scaffold for accelerating in vivo bone regeneration, holding great promise in treating orthopedic diseases.

The Effects of Graphene on the Biocompatibility of a 3D-Printed Porous Titanium Alloy

3D-printed titanium (Ti) materials have attracted much attention in the field of bone tissue repair. However, the combination strength of traditional alloy materials with bone tissue is lower, and the elastic modulus is higher than that of natural bone tissue, which makes the titanium alloy susceptible to stress shielding phenomena after implantation. Therefore, it is urgent to find better surface modification technology. In this study, the physical and chemical properties, toxicity, and proliferation of adipose stem cells of composite graphene-coated titanium alloy (Gr–Ti) were investigated using 3D-printed titanium alloy as a material model. Physical and chemical property tests confirmed that 3D printing could produce porous titanium alloy materials; the compressive strength and elastic modulus of the titanium alloy scaffolds were 91 ± 3 MPa and 3.1 ± 0.4 GPa, matching the elastic modulus of normal bone tissue. The surface characterization shows that graphene can be coated on titanium alloy by a micro-arc oxidation process, which significantly improves the surface roughness of titanium alloy. The roughness factor (Ra) of the Ti stent was 4.95 ± 1.12 μm, while the Ra of the Gr–Ti stent was 6.37 ± 0.72 μm. After the adipose stem cells were co-cultured with the scaffold for 4 h and 24 h, it was found that the Gr–Ti scaffold could better promote the early cell adhesion. CCK-8 tests showed that the number of ADSCs on the G–Ti scaffold was significantly higher than that on the Ti scaffold (p &lt; 0.01). The relative growth rate (RGR) of ADSCs in Gr–Ti was grade 0–1 (non-toxic). In the in vivo experiment of repairing a critical bone defect of a rabbit mandible, the bone volume fraction in the Gr–Ti group increased to 49.42 ± 3.28%, which was much higher than that in the Ti group (39.76 ± 3.62%) (p &lt; 0.05). In conclusion, the porous graphene–titanium alloy promotes the proliferation and adhesion of adipose stem cells with multidirectional differentiation potential, which has great potential for the application of bone tissue engineering in repairing bone defects in the future.