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Objective Carbon fiber-reinforced polyetheretherketone (CF/PEEK) possesses the elastic modulus similar to that of natural bone, yet its inherent surface hydrophobicity and bio-inertness restrict its application in oral implantation. To overcome this limitation, this study proposes the layer-by-layer surface functionalization strategy to construct the multifunctional CF/PEEK-based composite that integrates long-term antibacterial activity with synergistic osteogenic functions. Methods After sulfonation of CF/PEEK with 98% H₂SO₄, the polydopamine-assisted nanohydroxyapatite (PDA-nHA) intermediate layer and the zinc-loaded carboxymethyl chitosan (Zn/CMCS) coating were sequentially constructed, yielding the SCP-PHA-Zn/CMCS composite. A series of Characterizations including surface morphology, in vitro mineralization, antibacterial evaluation, cytocompatibility, osteogenic property, and osteogenesis-related gene mRNA expression levels were performed to assess in vitro biocompatibility and osteogenic performance. Results SEM showed SCP-PHA-Zn/CMCS possessed the 3D porous network, with uniformly deposited PDA-nHA presenting a fibrous network after Zn-CMCS wrapping. After SBF immersion, spherical apatite particles deposited on all samples. Water contact angles of SCP-PHA and SCP-PHA-Zn/CMCS were significantly lower than SCF/PEEK. SCP-PHA-Zn/CMCS significantly inhibited Staphylococcus aureus and Escherichia coli . All groups showed good biocompatibility and long-term biosafety. rBMSCs on SCP-PHA-Zn/CMCS showed the highest adhesion density, proliferation, ALP activity, matrix mineralization, and osteogenesis-related gene mRNA expression levels, significantly outperforming the other groups and demonstrating the best in vitro osteogenic capacity. Conclusion To address the surface hydrophobicity and bioinertia of CF/PEEK, this study successfully constructed the multifunctional SCP-PHA-Zn/CMCS composite using sulfonation pretreatment and the layer-by-layer functionalization assembly strategy. The obtained material exhibited favorable physicochemical properties, excellent biocompatibility and antibacterial performance, and significantly promoted the adhesion and proliferation of rBMSCs, enhanced ALP activity, matrix mineralization, and osteogenesis-related gene mRNA expression levels, demonstrating outstanding in vitro osteogenic capacity. These findings provide experimental evidence and a novel design strategy for the clinical translation of CF/PEEK in oral implantation and bone defect repair. Download: Download high-res image (282KB) Download: Download full-size image
Osteonecrosis is a devastating orthopedic disease in clinic that generally occurs in the femoral head associating with corticosteroid use up to 49 % in patients. In particular, glucocorticoids induced osteonecrosis of the femoral head is closely related to the local immune response that characterized by abnormal macrophage activation and inflammatory cell infiltration at the necrotic site, forming a pro-inflammatory microenvironment dominated by M1 macrophages, and thus leads to failure of bone repair and regeneration. Here, we report a bone regeneration strategy that constructs an immune regulatory biomaterial platform using an injectable thiolated hyaluronic acid hydrogel with lithium-doped nano-hydroxyapatite (Li-nHA@Gel) delivery for osteonecrosis treatment. Li-nHA@Gel achieved a sustain and longterm release of Li ions, which might enhance M2 macrophage polarization through the activation of the JAK1/STAT6/STAT3 signaling pathway, and the following induced pro-repair immune microenvironment mediated the enhancement of the osteogenic and angiogenic differentiation. Moreover, both in vitro and in vivo studies indicated that Li-nHA@Gel enhanced M2 macrophage polarization, osteogenesis, and angiogenesis, and thus promoted the bone and blood vessel formation. Taken together, this novel bone immunomodulatory biomaterial platform that promotes bone regeneration by enhancing M2 macrophage polarization, osteogenesis, and angiogenesis could be a promising strategy for osteonecrosis treatment.
Background The use of autologous bone grafting is considered the most successful method for managing bone defects, particularly when utilizing cancellous bone grafts for the best outcomes. Nonetheless, the scarcity of cancellous bone presents a notable obstacle in remedying these defects. Consequently, it is essential to create reliable alternatives to cancellous bone grafts to ensure effective management of bone defects.Methods In this research, we created an injectable composite hydrogel stents using gelatin methacrylate (GelMA) hydrogel to mimic the collagen properties of cancellous bone, along with the inclusion of nanohydroxyapatite (nHA) to signify the inorganic element. Furthermore, we incorporated vascular endothelial growth factor (VEGF) to improve regenerative vascular capabilities. Before being implanted into rat cranium defect models, these composite hydrogel stents were co-cultured with human umbilical vein endothelial cells (HUVEC) and bone marrow mesenchymal stem cells (BMSC).Results The composite hydrogel stents exhibited a network structure with porosity, robust mechanical properties, and beneficial degradation traits. In the degradation phase, it steadily releases Ca²⁺ and VEGF, which encourages the proliferation, migration, and osteogenic differentiation of BMSCs from rats. Moreover, this release improves the ability of HUVECs to form tubes. Collectively, these mechanisms support the regeneration of blood vessels and bone in the cranium defect region of rats.Conclusion The composite hydrogel stents demonstrated excellent cytocompatibility and biological characteristics, as evidenced by its ability to enhance both osteogenesis and angiogenesis in vivo and in vitro. Consequently, it has the potential to act as an effective alternative to natural cancellous bone.
The regenerative repair of segmental bone defect (SBD) is an urgent problem in the field of orthopedics. Rapid induction of angiogenesis and osteoinductivity after implantation of scaffold is critical. In this study, a unique tissue engineering strategy with mixture of peripheral blood-derived mesenchymal stem cells (PBMSC) and endothelial progenitor cells (PBEPC) was applied in a 3D-printed biphasic calcium phosphate (BCP) scaffold with highly bioactive nano hydroxyapatite (nHA) coating (nHA/BCP) to construct a novel vascularized tissue engineered bone (VTEB) for rabbit femoral SBD repair. The 2D coculture of PBMSC and PBEPC showed that they could promote the osteogenic or angiogenic differentiation of the cells from each other, especially in the group of PBEPC/PBMSC = 75:25. Besides, the 3D coculture results exhibited that the nHA coating could further promote PBEPC/PBMSC adhesion, proliferation, and osteogenic and angiogenic differentiation on the BCP scaffold. In vivo experiments showed that among the four groups (BCP, BCP-PBEPC/PBMSC, nHA/BCP, and nHA/BCP-PBEPC/PBMSC), the nHA/BCP-PBEPC/PBMSC group induced the best formation of blood vessels and new bone and, thus, the good repair of SBD. It revealed the synergistic effect of nHA and PBEPC/PBMSC on the angiogenesis and osteogenesis of the BCP scaffold. Therefore, the construction of VTEB in this study could provide a possibility for the regenerative repair of SBD.
Although artificial polymeric scaffolds act as vital characters in bone repair, their application is limited due to their inferior bioactivity. Herein, osteoinductive poly(ε-caprolactone) (PCL) composite scaffolds were prepared by synchronously enlisting bioactive nanohydroxyapatite (nHA), bioglass (BG), and bone morphogenetic protein-2 (BMP-2), which was bound up with polydopamine (pDA). It was found that pDA deposition not only significantly enhanced hydrophilicity and cell affinity of composite scaffolds but also endowed steady immobilization of BMP-2 with long-term yet sustained release. Compared to pure PCL and PCL/nHA/BG (PHB) scaffolds, the designed PHB-pDA-BMP-2 scaffolds significantly induced the differentiation of bone marrow stromal cells toward an osteogenic lineage. Meanwhile, in vivo examinations revealed the prominent osteogenic capability of PHB-pDA-BMP-2 scaffolds, which enabled complete bone healing of calvarial bone defects in rabbits by 12 weeks. This finding indicates that the developed porous composite scaffolds hold great potential for bone regeneration.
Addressing the clinical challenge of spatiotemporal coordination between vascularization and osteogenesis in bone repair, this study develops a composite scaffold system integrating piezoelectric stimulation and controlled drug release to achieve sequential regulation of angiogenesis-osteogenesis coupling. Combining piezoelectric barium titanate/nano-hydroxyapatite/polycaprolactone (BTO/nHA/PCL, BHP) with metformin (Met)-loaded GelMA (G@Met) hydrogels, the BHP/G@Met scaffolds are fabricated through dual-nozzle 3D printing. Material characterization confirms hierarchical porosity, stable piezoelectric output under ultrasound (US), and sustained drug release kinetics aligned with early-phase vascularization requirements. In vitro co-culture models demonstrate enhanced endothelial cell migration and tubular network formation during the initial 7d phase, followed by potent osteogenic differentiation of mesenchymal stem cells at intermediate (14d) and terminal (21d) periods. Dual-animal models reveal superior bone regeneration with extensive neovascularization and osseous tissue infiltration. Transcriptome sequencing and network pharmacological analyses identify PI3K-AKT signaling as the central pathway for BHP/G@Met coordinating sequential angiogenesis-osteogenesis. By synergizing piezoelectric bioelectrical cues with spatiotemporal drug delivery, BHP/G@Met pioneers a chronologically adaptive strategy to match the spatial-temporal bone healing microenvironments, offering a paradigm for dynamic tissue regeneration.
Barrier membranes have broad application prospects in guided bone regeneration (GBR), but their use in orthopedic and dental applications is severely limited by the germ-laden microenvironment and soft-hard tissue hierarchy. Thus, a novel nanocomposite multifunctionalized sandwich-like GBR membrane (SGM) with enhanced osteogenic properties and antimicrobial activities was fabricated by the sequential electrospinning of zinc oxide nanoparticle-doped silk fibroin (nZnO/SF), pure SF, and hydroxyapatite nanoparticle-doped SF (nHA/SF). SEM images demonstrated a well-developed porous three-dimensional sandwich-like hierarchical structure, while TEM, FTIR, XRD and EDX analyses confirmed that nZnO and nHA were successfully incorporated into the SF nanofibers. The water contact angle test indicated that the SGM exhibited satisfactory hydrophilicity after integration with nZnO and nHA. In vitro biological experiments proved that SGM had good cytocompatibility with MC3T3-E1 and L929 cells and promoted the differentiation and mineralization of MC3T3-E1 cells. Additionally, antibacterial assays suggested that the SGM exhibited noticeable antimicrobial activity in vitro and in vivo . Furthermore, the SGM promoted bone formation in both rat calvarial defect models and periodontal defect models. Consequently, the multifunctionalized nanofibrous SGM, which simultaneously exhibits good biocompatibility and enhanced osteogenic and antimicrobial properties, holds promise for GBR applications.
Preserving the vital pulp poses significant challenges in clinical pulp injury repair. Tricalcium silicate (C3S)-based cement has emerged as a successful pulp capping biomaterial. However, its efficacy in inducing pulp-dentin complex regeneration requires enhancement, and handling difficulties, poor anti-collapsible ability and inadequate self-setting performance remain significant obstacles, restricting its broader application in endodontics. In this study, a multifunctional SrCO 3 -doped C3S biocement (C3S-Sr/F127-nHA) reinforced by Pluronic® F127 thermoreversible hydrogel and nano hydroxyapatite (nHA) was developed to overcome the limitations of C3S-based cements. The thermoresponsive characteristic of F127 conferred the C3S pastes smart response behavior, allowing for optimal operability at room temperature and maintaining excellent anti-collapse properties at body temperature. C3S-Sr/F127-nHA exhibited an injectability rate of up to 93 %. nHA facilitated the hydration kinetics of C3S, resulting in reduced setting time and enhanced compressive strength. Furthermore, C3S-Sr/F127-nHA demonstrated excellent apatite mineralization ability, cytocompatibility, and potential for osteo/odontogenic induction. The inclusion of Sr 2+ ions and nHA promoted osteo/odontogenic differentiation of stem cells isolated from human exfoliated deciduous teeth (SHED). In vivo findings indicated that C3S-Sr/F127-nHA could stimulate the formation of a homogeneous dentin bridge. Consequently, C3S-Sr/F127-nHA is a promising biocement for use in regenerative endodontics.
The repair of large bone defects remains a challenging problem in bone tissue engineering. Ischemia and hypoxia in the bone defect area make it difficult for seed cells to survive and differentiate, which fail to perform effective tissue regeneration. Current oxygen-producing materials frequently encounter problems such as a rapid degradation rate, insufficient mechanical properties, difficult molding, and cumbersome fabrication. Here, a novel three-dimensional (3D) printed integrated bionic oxygenated scaffold was fabricated with gelatin-CaO2 microspheres, polycaprolactone (PCL), and nanohydroxyapatite (nHA) using low-temperature molding 3D printing technology. The scaffold had outstanding mechanical properties with bionic hierarchical porous structures. In vitro reports showed that the scaffold exhibited excellent cytocompatibility and could release O2 sustainably for more than 2 weeks, which significantly enhanced the survival, growth, and osteogenic differentiation of bone marrow mesenchymal stem cells under hypoxia. In vivo experiments revealed that the scaffold facilitated efficient bone repair after it was transplanted into a rabbit calvarial defect model. This result may be due to the scaffolds reducing hypoxia-inducible factor-1α accumulation, improving the expression of osteogenic regulatory transcription factors, and accelerating osteogenesis. In summary, the integrated bionic PCL/nHA/CaO2 scaffold had excellent capabilities in sustainable O2 release and bone regeneration, which provided a promising clinical strategy for bone defect repair.
Given the widespread clinical demand, addressing irregular cranial bone defects poses a significant challenge following surgical procedures and traumatic events. In situ-formed injectable hydrogels are attractive for irregular bone defects due to their ease of administration and the ability to incorporate ceramics, ions, and proteins into the hydrogel. In this study, a multifunctional hydrogel composed of oxidized sodium alginate (OSA)-grafted dopamine (DO), carboxymethyl chitosan (CMCS), calcium ions (Ca2+), nanohydroxyapatite (nHA), and magnesium oxide (MgO) (DOCMCHM) was prepared to address irregular cranial bone defects via dynamic Schiff base and chelation reactions. DOCMCHM hydrogel exhibits strong adhesion to wet tissues, self-healing properties, and antibacterial characteristics. Biological evaluations indicate that DOCMCHM hydrogel has good biocompatibility, in vivo degradability, and the ability to promote cell proliferation. Importantly, DOCMCHM hydrogel, containing MgO, promotes the expression of osteogenic protein markers COL-1, OCN, and RUNX2, and stimulates the formation of new blood vessels by upregulating CD31. This study could provide meaningful insights into ion therapy for the repair of cranial bone defects.
In bone tissue, nerves are primarily located in the periosteum and play an indispensable role in bone defect repair. However, most bone tissue engineering approaches ignored the reconstruction of the nerve network. Herein, we aimed to develop a bilayer hydrogel simulating periosteum-bone structure to induce innervated bone regeneration. The bottom “bone” layer consisted of gelatin methacryloyl (GelMA), poly (ethylene glycol) diacrylate (PEGDA), and nano-hydroxyapatite (nHA), whereas the upper “periosteum” layer consisted of GelMA, sodium alginate (SA) and MgCl2. The mechanical properties of the upper and bottom hydrogels were designed to be suitable for neurogenesis and osteogenesis, respectively. Besides, Mg2+ from the “periosteum” layer released at the early stage (within 7 d), which aligned with the optimal time window for nerve regeneration and osteogenic related neuropeptide release. Simultaneously, the prevention of long-term Mg2+ release (after 7 d) could avoid osteogenic inhibition caused by prolonged Mg2+ exposure. Additionally, the incorporation of nHA in the bottom “bone” layer supported the long-term osteogenesis due to its osteoconductivity and slow degradation. In vitro biological experiments revealed that the bilayer hydrogel (GS@Mg/GP@nHA) promoted neurite growth and calcitonin gene-related peptide (CGRP) expression in rat dorsal root ganglion (DRG) neurons, as well as the osteogenesis of rat bone-derived mesenchymal stem cells (BMSCs). Moreover, the in vivo experiments demonstrated that GS@Mg/GP@nHA hydrogel efficiently promoted nerve network reconstruction and bone regeneration of rat calvarial bone defects. Altogether, the bilayer hydrogel GS@Mg/GP@nHA could promote innervated bone regeneration, providing new insights for biomaterial designing for bone tissue engineering.
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