Femoral Defects Research Articles

Fibromodulin facilitates the osteogenic effect of Masquelet's induced membrane by inhibiting the TGF-β/SMAD signaling pathway.

Masquelet's induced membrane (IM) technique is a promising treatment strategy for the repair of substantial bone defects. The formation of an IM around polymethylmethacrylate bone cement plays a crucial role in this technique. Several studies have indicated that IMs have bioactivity because they contain abundant blood vessels, a variety of cells, and biological factors. The bioactivity of an IM increases during the initial stages of formation, thereby facilitating bone regeneration and remodeling. Nevertheless, the precise mechanisms underlying the enhancement of IM bioactivity and the promotion of bone regeneration necessitate further investigation. In this study, we successfully developed a Masquelet IM model of critical femur defects in rats. By employing proteomics analysis and biological detection techniques, we identified fibromodulin (FMOD) as a pivotal factor contributing to angiogenesis and the enhanced bioactivity of the IM. A significant increase in angiogenesis and the expression of bioactive factors in the IM was also observed with the upregulation of FMOD expression. Furthermore, this effect is mediated through the inhibition of the transforming growth factor beta (TGF-β)/SMAD signaling pathway. We also demonstrated that administering recombinant human FMOD enhanced osteogenesis in rat bone marrow mesenchymal stem cells and angiogenesis in human umbilical vein endothelial cells in vitro. Furthermore, the negative regulatory effect of the TGF-β signaling pathway was verified. In conclusion, this study provides a novel theoretical basis for the application of IMs in bone-defect reconstruction and explores possible new mechanisms that may play an important role in promoting the bioactivity and osteogenic potential of IMs.

Combination of Adipose-Derived Stromal/Stem Cells and Decellularized Adipose Tissue Hydrogel for Osteogenic Applications.

Adipose-derived stromal/stem cells (ASCs) and decellularized adipose tissue (DAT) are adipose tissue products obtained from individuals undergoing fat removal procedures like liposuction, lipectomy, or breast reduction. DAT hydrogel is prepared by removing the cells from the adipose tissue and digesting it to form a liquid material that forms a gel at physiological temperature. ASCs seeded on DAT have displayed osteogenic potential in vitro and in animal models of bone defects. Herein, we describe the methods for preparing DAT hydrogel, ASC seeding in DAT hydrogel, osteogenic differentiation of ASCs, creation of critical-sized femur defect model in mice, its treatment with ASC-DAT hydrogel, and analyses.

Combination of a human articular cartilage-derived extracellular matrix scaffold and microfracture techniques for cartilage regeneration: A proof of concept in a sheep model

BackgroundThe utilization of decellularized extracellular matrix has gained considerable attention across numerous areas in regenerative research. Of particular interest is the human articular cartilage-derived extracellular matrix (hACECM), which presents as a promising facilitator for cartilage regeneration. Concurrently, the microfracture (MF) ​technique, a well-established marrow stimulation method, has proven efficacious in the repair of cartilage defects. However, as of the current literature review, no investigations have explored the potential of a combined application of hACECM and the microfracture technique in the repair of cartilage defects within a sheep model. HypothesisThe combination of hACECM scaffold and microfracture will result in improved repair of full-thickness femoral condyle articular cartilage defects compared to the use of either technique alone. Study designControlled laboratory study. MethodsFull-thickness femoral condyle articular cartilage defect (diameter, 7.0 ​mm; debrided down to the subchondral bone plate) were created in the weight-bearing area of the femoral medial and lateral condyles (n ​= ​24). All of defected sheep were randomly divided into four groups: control, microfracture, hACECM scaffold, and hACECM scaffold ​+ ​microfracture. After 3, 6 and 12 months, the chondral repair was assessed for standardized (semi-) quantitative macroscopic, imaging, histological, immunohistochemical, mechanics, and biochemical analyses in each group. ResultAt 3, 6 and 12 months after implantation, the gross view and pathological staining of regenerative tissues were better in the hACECM scaffold and hACECM scaffold ​+ ​microfracture groups than in the microfracture and control groups; Micro-CT result showed that the parameters about the calcified layer of cartilage and subchondral bone were better in the hACECM scaffold and hACECM scaffold ​+ ​microfracture groups than the others, and excessive subchondral bone proliferation in the microfracture group. The results demonstrate that human cartilage extracellular matrix scaffold alone is an efficient, safe and simple way to repair cartilage defects. ConclusionhACECM scaffolds combined with/without microfracture facilitate chondral defect repair. The translational potential of this articlePreclinical large animal models represent an important adjunct and surrogate for studies on articular cartilage repair, while the sheep stifle joint reflects many key features of the human knee and are therefore optimal experimental model for future clinical application in human. In this study, we developed a human articular cartilage-derived extracellular matrix scaffold and to verify the viability of its use in sheep animal models. Clinical studies are warranted to further quantify the effects of hACECM scaffolds in similar settings.

New insight into biodegradable macropore filler on tuning mechanical properties and bone tissue ingrowth in sparingly dissolvable bioceramic scaffolds

Structural parameters of the implants such as shape, size, and porosity of the pores have been extensively investigated to promote bone tissue repair, however, it is unknown how the pore interconnectivity affects the bone growth behaviors in the scaffolds. Herein we systematically evaluated the effect of biodegradable bioceramics as a secondary phase filler in the macroporous networks on the mechanical and osteogenic behaviors in sparingly dissolvable bioceramic scaffolds. The pure hardystonite (HT) scaffolds with ∼550 & 800 μm in pore sizes were prepared by digital light processing, and then the Sr-doped calcium silicate (SrCSi) bioceramic slurry without and with 30 % organic porogens were intruded into the HT scaffolds with 800 μm pore size and sintered at 1150 °C. It indicated that the organic porogens could endow spherical micropores in the SrCSi filler, and the invasion of the SrCSi component could not only significantly enhance the compressive strength and modulus of the HT-based scaffolds, but also induce osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). The pure HT scaffolds showed extremely slow bio-dissolution in Tris buffer after immersion for 8 weeks (∼1 % mass decay); in contrast, the SrCSi filler would readily dissolve into the aqueous medium and produced a steady mass decay (>6 % mass loss). In vivo experiments in rabbit femoral bone defect models showed that the pure HT scaffolds showed bone tissue ingrowth but the bone growth was impeded in the SrCSi-intruded scaffolds within 4 weeks; however, the group with higher porosity of SrCSi filler showed appreciable osteogenesis after 8 weeks of implantation and the whole scaffold was uniformly covered by new bone tissues after 16 weeks. These findings provide some new insights that the pore interconnectivity is not inevitable to impede bone ingrowth with the prolongation of implantation time, and such a highly biodegradable and bioactive filler intrusion strategy may be beneficial for optimizing the performances of scaffolds in bone regenerative medicine applications.

Poly-ε-Caprolactone 3D-Printed Porous Scaffold in a Femoral Condyle Defect Model Induces Early Osteo-Regeneration.

Large bone reconstruction following trauma poses significant challenges for reconstructive surgeons, leading to a healthcare burden for health systems, long-term pain for patients, and complex disorders such as infections that are difficult to resolve. The use of bone substitutes is suboptimal for substantial bone loss, as they induce localized atrophy and are generally weak, and unable to support load. A combination of strong polycaprolactone (PCL)-based scaffolds, with an average channel size of 330 µm, enriched with 20% w/w of hydroxyapatite (HA), β-tricalcium phosphate (TCP), or Bioglass 45S5 (Bioglass), has been developed and tested for bone regeneration in a critical-size ovine femoral condyle defect model. After 6 weeks, tissue ingrowth was analyzed using X-ray computed tomography (XCT), Backscattered Electron Microscopy (BSE), and histomorphometry. At this point, all materials promoted new bone formation. Histological analysis showed no statistical difference among the different biomaterials (p > 0.05), but PCL-Bioglass scaffolds enhanced bone formation in the center of the scaffold more than the other types of materials. These materials show potential to promote bone regeneration in critical-sized defects on load-bearing sites.

Internal and external co-induction pineal 3D printed scaffolds for bone and blood vessel regeneration

The precise structural design and reproducible manufacturing advantages of the 3D printed scaffold make it attract attention in clinical applications. However, the inability of scaffolds to achieve internal and external co-induced vascularized osteogenesis limits their application. After observing the ingenious and functionalized structural combination of "pinecone", this study prepared hydrogel microspheres encapsulating strontium ranelate (SrR)-dendrimer (PAMAM) as a functionalized "pine nuts" through microfluidic technology. The 3D-printed Polycaprolactone (PCL) scaffold was used as a framework in which hydrogel microspheres and a 3D-printed scaffold were cleverly combined. In this pinecone 3D-scaffold system, the slow release of SrR is beneficial to promote vascularization and osteogenic differentiation inside and outside the scaffold. Furthermore, the rat femoral defect model verified that the pinecone scaffold promoting the formation of internal vascular network, osteogenic differentiation and shortening the bone repair time in vivo. In summary, this pinecone degradable biomimetic composite scaffold with internal osteogenic differentiation and vascular activation functions has great potential for clinical demand in segmental bone defects.

Fibrous topology promoted pBMP2-activated matrix on titanium implants boost osseointegration.

Titanium (Ti) implants have been extensively used after surgical operations. Its surface bioactivity is of importance to facilitate integration with surrounding bone tissue, and ultimately ensure stability and long-term functionality of the implant. The plasmid DNA-activated matrix (DAM) coating on the surface could benefit osseointegration but is still trapped by poor transfection for further application, especially on the bone marrow mesenchymal stem cells (BMSCs) in vivo practical conditions. Herein, we constructed a DAM on the surface of fibrous-grained titanium (FG Ti) composed of phase-transition lysozyme (P) as adhesive, cationic arginine-rich lipid (RLS) as the transfection agent and plasmid DNA (pDNA) for bone morphology protein 2 (BMP2) expression. The cationic lipid RLS improved up to 30-fold higher transfection than that of commercial reagents (Lipofectamine 2000 and polyethyleneimine) on MSC. And importantly, Ti surface topology not only promotes the DAM to achieve high transfection efficiency (∼75.7% positive cells) on MSC due to the favorable combination but also reserves its contact induction effect for osteoblasts. Upon further exploration, the fibrous topology on FG Ti could boost pDNA uptake for gene transfection, and cell migration in MSC through cytoskeleton remodeling and induce contact guidance for enhanced osteointegration. At the same time, the cationic RLS together with adhesive P were both antibacterial, showing up to 90% inhibition rate against Escherichia coli and Staphylococcus aureus with reduced adherent microorganisms and disrupted bacteria. Finally, the FG Ti-P/pBMP2 implant achieved accelerated bone healing capacities through highly efficient gene delivery, aligned surface topological structure and increased antimicrobial properties in a rat femoral condylar defect model.

Scaffold-Free Bone Marrow-Derived Mesenchymal Stem Cell Sheets Enhance Bone Formation in a Weight-Bearing Rat Critical Bone Defect Model.

Researchers have been exploring alternative methods for bone tissue engineering, as current management of critical bone defects may be a significant challenge for both patient and surgeon with conventional surgical treatments associated with several potential complications and drawbacks. Recent studies have shown mesenchymal stem cell sheets may enhance bone regeneration in different animal models. We investigated the efficacy of implanted scaffold-free bone marrow-derived mesenchymal stem cell (BMSC) sheets on bone regeneration of a critical bone defect in a weight-bearing rat model. BMSCs were isolated from the femora of male Sprague-Dawley rats 5-6 weeks of age and cell sheets were produced on temperature-responsive culture dishes. Nine male Sprague-Dawley rats 6-8 weeks of age were utilized. A bilateral femoral critical bone defect was created with a bridge plate serving as internal fixation. One side was randomly selected and BMSC sheets were implanted into the bone defect (BMSC group), with the contralateral side receiving no treatment (control). Rats were anesthetized and radiographs were performed at 2-week intervals. At the 8-week time point, rats were euthanized, femurs harvested, and microcomputed tomography and histological analysis was performed. We found a statistically significant increase in new bone formation and bone volume fraction compared with the control. Histomorphometry analysis revealed a larger percent of newly formed bone and a higher total histological score. Our results suggest that scaffold-free BMSC sheets may be used in the management of large weight-bearing bone defects to complement a different surgical technique or as a standalone approach followed by internal fixation. However, further research is still needed.

Engineered Cell‐Laden Armor Unit‐Mimicking Bioceramic Granules for Bone Regeneration

AbstractBioceramic granules have been used as fillers for the repair of various bone defects, but structural instability leads to susceptibility to displacement or dislodgement in practice. Armor unit is a concrete block placed along the breakwater to resist waves and protect the dike. Inspired by this, an armor unit‐mimicking (AU) bioceramic granule by three‐dimentional (3D) printing is developed, and combine it with adipose‐derived stem cells (ADSCs) overexpressing fibroblast growth factor 23 (FGF23) to construct tissue‐engineered bone. In vitro results show that AU granules and FGF23 transfection synergistically promote osteogenic differentiation of ADSCs. For the repair of beagle femoral metaphyseal defects, the implanted AU granules are stacked and interlocked to form an impact‐resistant porous scaffold. With the assistance of ADSCsFGF23, AU granules exhibit enhanced new bone formation and host bone maintenance. Taken together, these findings demonstrate the potential of 3D‐printed AU granules in combination with FGF23‐overexpressing ADSCs as a novel bone void filler for ease of handling and enhanced bone regeneration.

Enzyme-Mineralized PVASA Hydrogels with Combined Toughness and Strength for Bone Tissue Engineering.

Enzymatic mineralization is an advanced mineralization method that is often used to enhance the stiffness and strength of hydrogels, but often accompanied by brittle behavior. Moreover, the hydrogel systems with dense networks currently used for enzymatic mineralization are not ideal materials for bone repair applications. To address these issues, two usual bone repair hydrogels, poly(vinyl alcohol) (PVA) and sodium alginate (SA), were selected to form a double-network structure through repeated freeze-thawing and ionic cross-linking, followed by enzyme mineralization. The results demonstrated that both enzymatic mineralization and double-network structure improved the mechanical and biological properties and even exhibited synergistic effects. The mineralized PVASA hydrogels exhibited superior comprehensive mechanical properties, with a Young's modulus of 1.03 MPa, a storage modulus of 103 kPa, and an equilibrium swelling ratio of 132%. In particular, the PVASA hydrogel did not suffer toughness loss after mineralization, with a high toughness value of 1.86 MJ/m3. The prepared hydrogels also exhibited superior biocompatibility with a cell spreading area about 13 times that of mineralized PVA. It also effectively promoted cellular osteogenic differentiation in vitro and further promoted the formation of new bone in the femur defect region in vivo. Overall, the enzyme-mineralized PVASA hydrogel demonstrated combined strength and toughness and great potential for bone tissue engineering applications.

Prevention of ankle joint deformities in children on the side of fibular graft intake for microsurgical transposition

BACKGROUND: Reconstructive operations of the musculoskeletal system in children may require the removal of a massive autograft for microsurgical bone grafting. The traditional donor site is the diaphyseal part of the fibula. In the late postoperative period, these patients may develop complications such as a valgus deformity of the ankle joint on the side of the donor fibular defect.
 AIM: To analyze the effectiveness of surgical methods for the prevention of hallux valgus of the ankle joint after free transplantation of a fragment of the fibula to replace limb-bone defects in children.
 MATERIALS AND METHODS: The treatment results of 11 patients aged 11–16 years (6 girls and 5 boys), in whom an autograft from the fibular diaphysis was used to replace defects in long tubular bones were analyzed. Two patients had a femoral defect as a result of previously transferred hematogenous osteomyelitis. A congenital false joint of the femur was found in 2 patients, bones of the lower leg in 6, and the ulna in 1. The distal fragment of the resected fibula was stabilized by an autograft from the iliac bone, in eight and three cases at the level of the diaphysis and metaphysis of the specified fragment, respectively. The size of the resected fragment in % of the total length of the fibula and the level of distal osteotomy were considered. The presence and magnitude of the proximal displacement of the fragment and the position of the ankle joint gap were evaluated at least 5 years after surgery.
 RESULTS: In this study, ≥5 years after the intervention, proximal displacement of the distal fragment of the fibula was absent in only one patient. In the remaining 10, the displacement value did not exceed 3.5 mm. Valgus deformity in the ankle joint of 5° from the initial position developed in two patients. Its progression was prevented by temporary hemiepiphysis of the distal epiphysis of the tibia using a 4.0-mm diameter spongiose screw, whereas in up to 16 months, the valgus deformity progression stopped and decreased to initial values.
 CONCLUSIONS: Fibular resection in the optimal variant should preserve the distal part of the bone as much as possible, and the stability of the ankle joint increases with synostosis of the shin bones in the distal diaphyseal section without bringing the bones closer together. When a clinically significant valgus deformity appears (5° from the initial position) with the preservation of the function of the distal growth zone of the tibia, the use of temporary transphyseal hemiepiphysiodesis of the tibia for correction is optimal.

Combination effect of Chinese kidney-tonifying granules and platelet-rich plasma gels on enhancing bone healing in rat models with femur defects

BackgroundThe traditional Chinese kidney-tonifying granules, known as Bushen Zhongyao Keli (BSZYKL), have been found to stimulate calcium salt deposition, enhance bone formation, and foster bone growth within the bone matrix at sites of bone defects. On the other hand, platelet-rich plasma (PRP) is enriched with various growth factors capable of facilitating the repair of bone defects and enhancing bone strength following fractures. This study is dedicated to investigating the combined efficacy of BSZYKL and PRP gel (PRP-G) in the treatment of bone defects.MethodsWe established a femur defect model in male Sprague-Dawley (SD) rats and filled the defect areas with autologous coccygeal bone and PRP-G. For 8 consecutive weeks, those rats were given with intragastric administration of BSZYKL. Biomechanical characteristics of the femur were assessed 28 days after intramuscular administration. On day 56, bone formation was examined using X-ray, micro-CT, and transmission electron microscopy. Additionally, we analyzed the expression of bone formation markers, Runx2 and Osterix, in femur tissues through qPCR, Western blotting, and immunohistochemistry.ResultsRats receiving the combined treatment of BSZYKL and PRP-G exhibited drastically enhanced femoral peak torsion, failure angle, energy absorption capacity, and torsional stiffness as compared to control group. This combination therapy also led to marked improvements in bone volume, mass, and microarchitecture, accompanied by elevated expressions of Runx2 and Osterix when compared to control group. Notably, the synergistic effects of BSZYKL and PRP-G in treating bone defects surpassed the effects of either treatment alone.ConclusionsThese findings revealed the potential of BSZYKL in combination with PRP-G in improving bone defects.

Advances in revision surgery after primary total hip arthroplasty for Crowe type Ⅳ developmental dysplasia of the hip

To review research advances of revision surgery after primary total hip arthroplasty (THA) for patients with Crowe type Ⅳ developmental dysplasia of the hip (DDH). The recent literature on revision surgery after primary THA in patients with Crowe type Ⅳ DDH was reviewed. The reasons for revision surgery were analyzed and the difficulties of revision surgery, the management methods, and the related prosthesis choices were summarized. Patients with Crowe type Ⅳ DDH have small anteroposterior diameter of the acetabulum, large variation in acetabular and femoral anteversion angles, severe soft tissue contractures, which make both THA and revision surgery more difficult. There are many reasons for patients undergoing revision surgery after primary THA, mainly due to aseptic loosening of the prosthesis. Therefore, it is necessary to restore anatomical structures in primary THA, as much as possible and reduce the generation of wear particles to avoid postoperative loosening of the prosthesis. Due to the anatomical characteristics of Crowe type Ⅳ DDH, the patients have acetabular and femoral bone defects, and the repair and reconstruction of bone defects become the key to revision surgery. The acetabular side is usually reconstructed with the appropriate acetabular cup or combined metal block, Cage, or custom component depending on the extent of the bone defect, while the femoral side is preferred to the S-ROM prosthesis. In addition, the prosthetic interface should be ceramic-ceramic or ceramic-highly cross-linked polyethylene wherever possible. The reasons leading to revision surgery after primary THA in patients with Crowe type Ⅳ DDH and the surgical difficulties have been clarified, and a large number of clinical studies have proposed corresponding revision modalities based on which good early- and mid-term outcomes have been obtained, but further follow-up is needed to clarify the long-term outcomes. With technological advances and the development of new materials, personalized prostheses for these patients are expected to become a reality.

An in silico micro-multiphysics agent-based approach for simulating bone regeneration in a mouse femur defect model.

Bone defects represent a challenging clinical problem as they can lead to non-union. In silico models are well suited to study bone regeneration under varying conditions by linking both cellular and systems scales. This paper presents an in silico micro-multiphysics agent-based (micro-MPA) model for bone regeneration following an osteotomy. The model includes vasculature, bone, and immune cells, as well as their interaction with the local environment. The model was calibrated by time-lapsed micro-computed tomography data of femoral osteotomies in C57Bl/6J mice, and the differences between predicted bone volume fractions and the longitudinal in vivo measurements were quantitatively evaluated using root mean square error (RMSE). The model performed well in simulating bone regeneration across the osteotomy gap, with no difference (5.5% RMSE, p = 0.68) between the in silico and in vivo groups for the 5-week healing period - from the inflammatory phase to the remodelling phase - in the volume spanning the osteotomy gap. Overall, the proposed micro-MPA model was able to simulate the influence of the local mechanical environment on bone regeneration, and both this environment and cytokine concentrations were found to be key factors in promoting bone regeneration. Further, the validated model matched clinical observations that larger gap sizes correlate with worse healing outcomes and ultimately simulated non-union. This model could help design and guide future experimental studies in bone repair, by identifying which are the most critical in vivo experiments to perform.

Chitosan-Reinforced Gelatin Microspheres-Modified Glass Ionomer Cement (GIC): A Novel Bone Alloplast Graft Material Synthesis and an In Vivo Analysis.

Aim and objective The study aimed to assess and evaluate the efficacyof glass ionomer modified with chitosan-reinforced gelatin microspheres on bone formation. Materials and methods The study involved three groups: Group I comprised plain glass ionomer cement; Group II comprised glass ionomer cement/gelatin (70:30 wt%); in Group III, glass ionomer cement/gelatin/chitosan (70:30%) scaffold were made into discs; the gelatin microspheres were synthesized by oil emulsion method. The synthesized scaffold was subjected to the following in vitro testing, Instron Universal Testing Machine (UTM), U3000, (Instron Corporation, Norwood, Massachusetts, United States) to assess compressive strength, scanning electron microscope(SEM) examination, and biocompatibility testing using hemocompatibility assay. The material was then testedin vivo; male Wistar albino rats, a total of nine animals, were utilized for this purpose. Three animals were used in each group; a femoral defect model was the model of choice for the experiment and the animals were observed for a period of four weeks, following which the animals were sacrificed and sent for histopathological analysis. Results The compression testing was carried out using UTM; test group I was 33 MPa, test group II was 2.3 MPa, and test group III was 25.75 MPa. SEM (JSM-IT800 Schottky Field Emission NANO SEM (JEOL, Tokyo, Japan)) analysis was done to observe the porosity of the fabricated scaffold with the average measurement of0.12 ± 0.2 μm in test group II and 0.29 ± 0.4μm in test group III. Hemocompatibility reports noted 0.4-0.8% lysis for the synthesized scaffolds. Histopathology staining of the femur defects showed that group III favoured bone formation. One-way analysis of variance (ANOVA) and post hoc Bonferroni test was done on the data. The optical density values of the alizarin red stained slide showed statistical significance for group III. Conclusion In conclusion, the synthesized scaffolds are biocompatible, distribution of porosity andpore characteristics in the glass ionomer cement/gelatin/chitosan group is better than that of the glass ionomer cement/gelatin group. The glass ionomer cement/gelatin/chitosan group had better compressive strength and induced more bone formation compared to the other test group and the control. Thus, the novel glass ionomer modified with chitosan-reinforced gelatin microspheres has optimal properties to be used as a bone graft material.

3D-printed porous PEI/TCP composite scaffolds loaded with graphdiyne oxide on the surface for bone defect repair and near-infrared light-responsive antibacterial

Bone implants that promote osseointegration and have antimicrobial capabilities are necessary to ensure optimal long-term functionality of implants. In this study, a porous composite osteogenic scaffold was prepared using a twin-screw extruder and a fused deposition modeling three-dimensional (3D) printer. A polydopamine (PDA) coating was used to load graphdiyne oxide (GDYO) on the surface of the 3D printed polyetherimide-tricalcium phosphate (PEI-TCP) osteogenic scaffold through π–π double-bond interactions. The resulting P-T-GDYO composite scaffold was characterized, and its mechanical properties, ability to promote osteogenic differentiation, and light-responsive antibacterial activity were evaluated. When the scaffold was doped with 10 wt% β-TCP, its mechanical properties remained in the range of those of cancellous bone. In terms of osteogenic differentiation, the P-T-GDYO composite scaffold promoted adhesion and osteogenic differentiation of MC3T3-E1 cells, as well as effectively promoting endogenous bone regeneration of rabbit femoral condyle bone defects in osteogenesis experiments in vivo. In a mouse subcutaneous infection model, after near-infrared light irradiation, P-T-GDYO showed an antibacterial rate of 90.18 % against Staphylococcus aureus, while maintaining good tissue–organ compatibility. This study represents an important development in GDYO-based photothermal antibacterial materials in the field of orthopedics and expands its application in the biomedical field.

A Multifunctional Scaffold for Bone Infection Treatment by Delivery of microRNA Therapeutics Combined With Antimicrobial Nanoparticles.

Treating bone infections and ensuring bone repair is one of the greatest global challenges of modern orthopedics, made complex by antimicrobial resistance (AMR) risks due to long-term antibiotic treatment and debilitating large bone defects following infected tissue removal. An ideal multi-faceted solution would will eradicate bacterial infection without long-term antibiotic use, simultaneously stimulating osteogenesis and angiogenesis. Here, a multifunctional collagen-based scaffold that addresses these needs by leveraging the potential of antibiotic-free antimicrobial nanoparticles (copper-doped bioactive glass, CuBG) to combat infection without contributing to AMR in conjunction with microRNA-based gene therapy (utilizing an inhibitor of microRNA-138) to stimulate both osteogenesis and angiogenesis, is developed. CuBG scaffolds reduce the attachment of gram-positive bacteria by over 80%, showcasing antimicrobial functionality. The antagomiR-138 nanoparticles induce osteogenesis of human mesenchymal stem cells in vitro and heal a large load-bearing defect in a rat femur when delivered on the scaffold. Combining both promising technologies results in a multifunctional antagomiR-138-activated CuBG scaffold inducing hMSC-mediated osteogenesis and stimulating vasculogenesis in an in vivo chick chorioallantoic membrane model. Overall, this multifunctional scaffold catalyzes killing mechanisms in bacteria while inducing bone repair through osteogenic and angiogenic coupling, making this platform a promising multi-functional strategy for treating and repairing complex bone infections.

Chitosan microcarriers deposited with Mg2+-doped phase-transited lysozyme: osteogenesis, pro-angiogenesis and anti-inflammatory for promoting bone regeneration

Microcarriers, as injectable cell carriers, provide a new strategy for bone defect repair due to their advantages of large specific surface area, flexible and controllable size, and injectability. Magnesium ion (Mg2+) is important trace element and has been demonstrated to improve bone tissue regeneration. The development of bone tissue engineering microcarriers that can sustainably control Mg2+ release could show significant impacts on bone regeneration. Herein, chitosan (CS) microcarriers deposited with Mg2+-doped phase-transited lysozyme (PTL) were fabricated for accelerating bone regeneration. The results prove that Mg2+-doped PTL was successfully self-assembled onto CS microcarriers through hydrogen bonds and electrostatic interactions. The introduction of Mg2+-doped PTL in CS microcarriers increased zeta potential of the materials. The microcarriers exhibited a spherical and interconnected porous structure with sizes of 300–400 μm and pore sizes of 35–40 μm. Due to the PTL deposition, the microcarriers displayed controlled release of Mg2+. Moreover, they showed good degradation ability. In vitro, the composite microcarriers supported cell attachment and improved proliferation and osteogenic differentiation of stem cells, attributed to the combined effects of PTL and Mg2+. Furthermore, they stimulated cell migration and tube formation of HUVECs, and induced LPS-activated macrophages polarized to the anti-inflammatory M2 phenotype. In vivo, the microcarriers accelerated bone regeneration, enhanced angiogenesis, and reduced inflammation in a rat model of right medial femoral malleolus defects. In summary, the results demonstrate that the prepared composite microcarriers have potential to be used as a new type of microcarrier for bone tissue engineering applications.

Functionalized biomimetic mineralized collagen promotes osseointegration of 3D-printed titanium alloy microporous interface

Mineralized collagen (MC) is the fundamental unit of natural bone tissue and can induce bone regeneration. Unmodified MC has poor mechanical properties and a single component, making it unable to cope with complex physiological environment. In this study, we introduced sodium alginate (SA) and vascular endothelial growth factor (VEGF) into the MC material to construct functionalized mineralized collagen (FMC) with good mechanical strength and the ability to continuously release growth factors. The FMC is filled into the pores of 3D printed titanium alloy scaffold to form a new organic-inorganic bioactive interface. With the continuous degradation of FMC, bone marrow mesenchymal stem cells (BMSCs) and vascular endothelial cells (VECs) in the surrounding environment are recruited to the surface of the scaffold to promote bone and vascular regeneration. After implanting the scaffold into the distal femoral defect of rabbits, Micro CT, histological, push-out, as well as immunohistochemical analysis showed that the composite interface can significantly promote osseointegration. These findings provide a new strategy for the development and application of mineralized collagen materials.

Development of artificial bone graft via in vitro endochondral ossification (ECO) strategy for bone repair

Endochondral ossification (ECO) is a form of bone formation whereby the newly deposited bone replaces the cartilage template. A decellularized artificial cartilage graft (dLhCG), which is composed of hyaline cartilage matrixes, has been developed in our previous study. Herein, the osteogenesis of bone marrow-derived MSCs in the dLhCG through chondrogenic differentiation, chondrocyte hypertrophy, and subsequent transdifferentiation induction has been investigated by simulating the physiological processes of ECO for repairing critical-sized bone defects. The MSCs were recellularized into dLhCGs and subsequently allowed to undergo a 14-day proliferation period (mrLhCG). Following this, the mrLhCG constructs were subjected to two distinct differentiation induction protocols to achieve osteogenic differentiation: chondrogenic medium followed by chondrocytes culture medium with a high concentration of fetal bovine serum (CGCC group) and canonical osteogenesis inducing medium (OI group). The formation of a newly developed artificial bone graft, ossified dLhCG (OsLhCG), as well as its capability of aiding bone defect reconstruction were characterized by in vitro and in vivo trials, such as mRNA sequencing, quantitative real-time PCR (qPCR), immunohistochemistry, the greater omentum implantation in nude mice, and repair for the critical-sized femoral defects in rats. The results reveal that the differentiation induction of MSCs in the CGCC group can realize in vitro ECO through chondrogenic differentiation, hypertrophy, and transdifferentiation, while the MSCs in the OI group, as expected, realize ossification through direct osteogenic differentiation. The angiogenesis and osteogenesis of OsLhCG were proved by being implanted into the greater omentum of nude mice. Besides, the OsLhCG exhibits the capability to achieve the repair of critical-size femoral defects.