PI3K/AKT/mTOR signaling regulates BCP ceramic-induced osteogenesis.

An increasing number of studies demonstrate that biphasic calcium phosphate (BCP) ceramics can induce bone regeneration. However, the underlying molecular mechanisms involved are still poorly understood. This work was proposed to investigate how PI3K/AKT/mTOR signaling influenced the osteogenesis mediated by BCP ceramics. The results showed that incubation with BCP ceramics promoted the proliferation of murine bone marrow-derived mesenchymal stem cells (BMSCs) in a time-dependent manner. The resulting cell proliferation was then suppressed by the selective inhibition of either PI3K, AKT, or mTOR signaling activation. Next, we confirmed that BCP ceramics up-regulated the phosphorylation levels of AKT and mTOR in BMSCs, suggesting the ability of BCP ceramics to drive the activation of PI3K/AKT/mTOR signaling in BMSCs. Furthermore, the blockade of PI3K/AKT/mTOR signaling prevented BCP ceramics-induced osteogenic differentiation and pro-angiogenesis of BMSCs by down-regulating the expression of genes encoding OPN, RUNX2 or VEGF. Moreover, the PI3K/AKT/mTOR signaling blockade suppressed stem cell infiltration and new bone formation in the implants following intra-muscular implantation of BCP ceramics in mice. Therefore, our results suggested that PI3K/AKT/mTOR signaling played a critical regulatory role in BCP ceramic-induced osteogenesis.

Bone Volume Analysis and Associated Influencing Factors in Mandibular Distraction Osteogenesis: A Retrospective Analysis.

BACKGROUND: Distraction osteogenesis is a progressively popular technique for maxillofacial bone reconstruction, but there is a notable gap in the analysis of bone volume within the distraction segment and the exploration of associated influencing factors. PURPOSE: The purpose of this study was to quantitatively analyze the new bone volume and the distraction gap volume in the three-dimensional (3D) model and explore the influencing factors associated with the percentage of the new bone volume to the distraction gap volume. STUDY DESIGN, SETTING, SAMPLE: This retrospective study included patients who underwent maxillofacial distraction osteogenesis treatment at the West China Hospital of Stomatology between 2014 and 2022, utilizing the mandibular distractor (Cebei, Ningbo, China). Exclusion criteria encompassed individuals with incomplete clinical or radiographical records as well as those who loss to follow-up. PREDICTOR VARIABLE: The predictor variables were age, sex, diagnosis, consolidation period duration, distraction modality, osteotomy area, distraction gap volume, and proximal bone segment volume. MAIN OUTCOME VARIABLE(S): The outcome variable was osteogenic effect which defined as the percentage of the new bone volume to the distraction gap volume in the 3D model. COVARIATES: Not applicable. ANALYSES: T-tests were used to describe categorical variables, and Pearson correlation analysis was used to describe continuous variables. Linear regression was employed to assess the predictiveness of variables for osteogenic effect. Data are described as mean ± standard deviation; statistical significance was established at a P value < .05. RESULTS: The study sample contained 35 patients(11 males and 24 females) with a mean age of 21.17 ± 11.82 years (range: 5 to 47 years) were included. The mean osteogenic effect of all samples was 78.89 ± 24.70%. Multiple linear regression models confirmed that the osteogenic effect was significantly influenced by the distraction gap volume (P = .003), proximal bone segment volume (P = .009), osteotomy area (P = .034), diagnosis (P = .004), and distraction modality (P = .021). CONCLUSION AND RELEVANCE: The percentage of new bone mass to simulated volume based on 3D model measurement can serve as an effective quantitative indicator for evaluating the osteogenic effect; our study demonstrates that distraction gap volume, proximal bone segment volume, osteotomy area, diagnosis, and distraction modality can statistically significantly influence the osteogenic effect.

Understanding the Mechanics of the Temporomandibular Joint Osteochondral Interface from Micro- and Nanoscopic Perspectives.

The condylar cartilage of the temporomandibular joint (TMJ) is connected to the subchondral bone by an osteochondral interface that transmits loads without causing fatigue damage. However, the microstructure, composition, and mechanical properties of this interface remain elusive. In this study, we found that structurally, a spatial gradient assembly of hydroxyapatite (HAP) particles exists in the osteochondral interface, with increasing volume of apatite crystals with depth and a tendency to form denser and stacked structures. Combined with nanoindentation, this complex assembly of nanoscale structures and components enhanced energy dissipation at the osteochondral interface, achieving a smooth stress transition between soft and hard tissues. This study comprehensively demonstrates the elemental composition and complex nanogradient spatial assembly of the osteochondral interface at the ultramicroscopic scale, providing a basis for exploring the construction of complex mechanical models of the interfacial region.

Micron/Submicron Scaled Hierarchical Ti Phosphate/Ti Oxide Hybrid Coating on 3D Printed Scaffolds for Improved Osteointegration.

Three-dimensional (3D) printed implants have attracted substantial attention in the field of personalized medicine, but negative impacts on mechanical properties or initial osteointegration have limited their application. To address these problems, we prepared hierarchical Ti phosphate/Ti oxide (TiP-Ti) hybrid coatings on 3D printed Ti scaffolds. The surface morphology, chemical composition, and bonding strength of the scaffolds were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), contact angle measurement, X-ray diffraction (XRD), and scratch test. In vitro performance was analyzed by colonization and proliferation of rat bone marrow mesenchymal stem cells (BMSCs). In vivo osteointegration of the scaffolds in rat femurs was assessed by micro-CT and histological analyses. The results demonstrated improved cell colonization and proliferation as well as excellent osteointegration obtained by incorporation of our scaffolds with the novel TiP-Ti coating. In conclusion, micron/submicron scaled Ti phosphate/Ti oxide hybrid coatings on 3D printed scaffolds have promising potential in future biomedical applications.

Decellularized-disc based allograft and xenograft prosthesis for the long-term precise reconstruction of temporomandibular joint disc.

Currently, no effective disc reconstruction treatment strategy is clinically available for temporomandibular joint (TMJ) disc-related diseases. To address this, we developed a prosthesis construct with laser-drilled decellularized natural disc reinforced by polycaprolactone, which mimics the natural morphology, and structural, biomechanical and biological property of the TMJ disc. The construct demonstrated good biocompatibility, safety and immunological tolerance both in vitro, and in a rat subcutaneous model. During 6 months implantation in an allogeneic rabbit TMJ disc reconstruction model, the disc prosthesis maintained its integrity, collagen fiber-orientation, mechanical property, joint structural stability and prevented articular cartilage and bone from damage. Furthermore, the "upgraded" disc prosthesis obtained from decellularized porcine disc was implanted into a goat TMJ disc reconstruction model. The xenograft prosthesis, with strength and viscoelasticity similar to a natural TMJ disc, was able to restore the structure and function of TMJ up to 20 weeks. These results demonstrate the translational feasibility of an allogeneic or xenogeneic decellularized disc prosthesis for treatment of advanced TMJ disc-related diseases. STATEMENT OF SIGNIFICANCE: This study makes a significant contribution to TMJ disc disease treatment both in theory and in clinics, because: (1) it provided an innovative approach to prepare an artificial TMJ disc with decent mechanical properties and long-term condyle-protecting effect; (2) it specified an advanced decellularized method for fibrocartilage decellularization and xenograft application; (3) it developed a facile and reproducible TMJ disc reconstruction model not only for middle size animal but also for large animal study; (4) the comprehensive and unreported biomechanical tests on the natural TMJ discs would act as a valuable reference for further research in the field of artificial TMJ disc materials or TMJ disc tissue engineering; (5) it suggested a potential treatment for patients with severe TMJ diseases that were commonly met but difficult to treat in clinics.

Canine ACL reconstruction with an injectable hydroxyapatite/collagen paste for accelerated healing of tendon-bone interface.

Healing of an anterior cruciate ligament (ACL) autologous graft in a bone tunnel occurs through the formation of fibrovascular scar tissue, which is structurally and compositionally inferior to normal fibrocartilaginous insertion and thus may increase the reconstruction failure and the rate of failure recurrence. In this study, an injectable hydroxyapatite/type I collagen (HAp/Col Ⅰ) paste was developed to construct a suitable local microenvironment to accelerate the healing of bone-tendon interface. Physicochemical characterization demonstrated that the HAp/Col Ⅰ paste was injectable, uniform and stable. The in vitro cell culture illustrated that the paste could promote MC3T3-E1 cells proliferation and osteogenic expression. The results of a canine ACL reconstruction study showed that the reconstructive ACL had similar texture and color as the native ACL. The average width of the tunnel, total bone volume, bone volume/tissue volume and trabecular number acquired from micro-CT analysis suggested that the healing of tendon-bone interface in experimental group was better than that in control group. The biomechanical test showed the maximal loads in experimental group achieved approximately half of native ACL's maximal load at 24 weeks. According to histological examination, Sharpey fibers could be observed as early as 12 weeks postoperatively while a typical four-layer transitional structure of insertion site was regenerated at 48 weeks in the experimental group. The injectable HAp/Col Ⅰ paste provided a biomimetic scaffold and microenvironment for early cell attachment and proliferation, further osteogenic expression and extracellular matrix deposition, and in vivo structural and functional regeneration of the tendon-bone interface.

Selective Polyetheretherketone Implants Combined with Graphene Cause Definitive Cell Adhesion and Osteogenic Differentiation.

Introduction: Polyetheretherketone (PEEK) has good biosafety and chemical stability for bone repair. However, PEEK is biologically inert and cannot promote bone apposition. This study investigated whether graphene-modified PEEK (G-PEEK) could improve cell adhesion and osteogenic differentiation. Methods: G-PEEK was prepared by melted blending and was characterized. In vitro, the biocompatibility of G-PPEK and the ability to promote cell adhesion and osteogenic differentiation in rabbit bone marrow mesenchymal stem cells (rBMSCs) were examined using live and dead cell double staining, the cell counting kit-8 (CCK-8) assay, immunofluorescence and quantitative real-time PCR (qRT‒PCR). An in vivo rabbit extra-articular graft-to-bone healing model was established. At 4 and 12 weeks after surgery, CT analysis and histological evaluation were performed. Results: In vitro, G-PEEK significantly improved the adhesion and proliferation of rBMSCs, with good biocompatibility. In vivo, G-PEEK promoted new bone formation at the site of the bone defect. Conclusion: G-PEEK showed excellent osteogenesis performance, which promises new applications in implant materials.

The biological effect of recombinant humanized collagen on damaged skin induced by UV-photoaging: An in vivo study.

The application of medical devices to repair skin damage is clinically accepted and natural polymer enjoys an important role in this field, such as collagen or hyaluronic acid, etc. However, the biosafety and efficacy of these implants are still challenged. In this study, a skin damage animal model was prepared by UV-photoaging and recombinant humanized type III collagen (rhCol III) was applied as a bioactive material to implant in vivo to study its biological effect, comparing with saline and uncrosslinked hyaluronic acid (HA). Animal skin conditions were non-invasively and dynamically monitored during the 8 weeks experiment. Histological observation, specific gene expression and other molecular biological methods were applied by the end of the animal experiment. The results indicated that rhCol III could alleviate the skin photoaging caused by UV radiation, including reduce the thickening of epidermis and dermis, increase the secretion of Collagen I (Col I) and Collagen III (Col III) and remodel of extracellular matrix (ECM). Although the cell-material interaction and mechanism need more investigation, the effect of rhCol III on damaged skin was discussed from influence on cells, reconstruction of ECM, and stimulus of small biological molecules based on current results. In conclusion, our findings provided rigorous biosafety information of rhCol III and approved its potential in skin repair and regeneration. Although enormous efforts still need to be made to achieve successful translation from bench to clinic, the recombinant humanized collagen showed superiorities from both safety and efficacy aspects.

Bioinspired Fabrication of Calcium-Doped TiP Coating with Nanofibrous Microstructure to Accelerate Osseointegration.

Bioinspired by the morphology of osteoclast-resorbed bone surfaces, we prepared a calcium-doped titanium phosphate (Ca-TiP) coating, which consists of a nanofibrous network, on titanium (Ti) substrate via a simple two-step hydrothermal method, trying to mimic natural bone compositionally and microstructurally. The in vitro studies show that the Ca-TiP coating with synergistic features of nanofibrous biomimetic topography and surface chemistry could elicit intensively osteogenic behavior and responses including enhanced cell adhesion, spreading, and proliferation as well as alkaline phosphatase (ALP) activity and up-regulated expression of bone-related genes, which inevitably benefit the formation of new bone and the quality of osseointegration. When the two control groups are compared in vivo, the significantly improved new bone formation in the early stage and the much stronger interfacial bonding with the surrounding bone for Ca-TiP coating suggest that Ca-TiP coating modified Ti implants hold great potential for orthopedic and dental applications.

Exploring the mechanism behind improved osteointegration of phosphorylated titanium implants with hierarchically structured topography.

Titanium (Ti) and its alloys have been frequently used in dental and orthopedic implants, but the undesired oxide layer easily formed on the surface tends to be the cause of implant failure for Ti-based implants. To address this problem, we herein prepared a phosphorylated Ti coating (TiP-Ti) with a micro/nano hierarchically structured topography on commercially pure Ti implants by a hydrothermal method to improve its osteointegration capacity. The surface morphology, chemical composition, and biological activity were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), contact-angle measurement, and protein adsorption assay. Osteointegration of TiP-Ti implants in rat tibia was investigated by biomechanical testing, micro-CT and histological analyses. We further explored the proposed mechanism which improves osteointegration of TiP-Ti implants by proliferation, adhesion, and differentiation assays of rat bone marrow mesenchymal stem cells (BMSCs). Our results demonstrated that the improved osteointegration mainly benefited from the better spread and adhesion of BMSCs on the micro/nano hierarchically structured TiP-Ti surfaces compared to hydroxyapatite coated Ti (HA-Ti), the positive control, and untreated Ti (untreated-Ti), the negative control. In conclusion, TiP-Ti surface is a promising candidate implant surface design to accelerate the osteointegration of Ti-based implants in biomedical applications.

The custom making of hierarchical micro/nanoscaled titanium phosphate coatings and their formation mechanism analysis.

In this study, a series of hierarchical micro/nanoscaled titanium phosphate (TiP) coatings possessing various surface morphologies were successfully fabricated on titanium (Ti) discs. The hydrothermal reactions of Ti discs in hydrogen peroxide (H2O2) and phosphoric acid (H3PO4) mixed solution yield diverse topographies such as hemispheric clump, cylindrical rod, spherical walnut, micro/nano grass, micro/nano sheet, and fibrous network. And their crystal structures were mainly composed of Ti(HPO4)2·0.5H2O, (TiO)2P2O7, H2TiP2O8, Ti(HPO4)2 and TiO2. The morphology and crystal shape of the TiP coatings depend strongly on the mass ratio of H2O2/H3PO4, reaction temperature and water content. Besides, the formation mechanism of TiP coatings with diverse morphologies was explored from the perspective of energetics and crystallography. The mechanism exploration paved the way for custom-making TiP coatings with desirable micro/nanoscaled morphologies to meet specific application purposes. The in vitro cytological performances of TiP coatings were also evaluated by co-culturing with rat bone marrow stromal cells (BMSCs), demonstrating a positive prospect for their use in bone tissue engineering.