Bone Defects In Animal Models Research Articles

MicroRNA-200c Release from Gelatin-Coated 3D-Printed PCL Scaffolds Enhances Bone Regeneration.

The fabrication of clinically relevant synthetic bone grafts relies on combining multiple biodegradable biomaterials to create a structure that supports the regeneration of defects while delivering osteogenic biomolecules that enhance regeneration. MicroRNA-200c (miR-200c) functions as a potent osteoinductive biomolecule to enhance osteogenic differentiation and bone formation; however, synthetic tissue-engineered bone grafts that sustain the delivery of miR-200c for bone regeneration have not yet been evaluated. In this study, we created novel, multimaterial, synthetic bone grafts from gelatin-coated 3D-printed polycaprolactone (PCL) scaffolds. We attempted to optimize the release of pDNA encoding miR-200c by varying gelatin types, concentrations, and polymer crosslinking materials to improve its functions for bone regeneration. We revealed that by modulating gelatin type, coating material concentration, and polymer crosslinking, we effectively altered the release rates of pDNA encoding miR-200c, which promoted osteogenic differentiation in vitro and bone regeneration in a critical-sized calvarial bone defect animal model. We also demonstrated that crosslinking the gelatin coatings on the PCL scaffolds with low-concentration glutaraldehyde was biocompatible and increased cell attachment. These results strongly indicate the potential use of gelatin-based systems for pDNA encoding microRNA delivery in gene therapy and further demonstrate the effectiveness of miR-200c for enhancing bone regeneration from synthetic bone grafts.

Advances in the Study of Extracellular Vesicles for Bone Regeneration.

Promoting the efficiency of bone regeneration in bone loss diseases is a significant clinical challenge. Traditional therapies often fail to achieve better therapeutic outcomes and shorter treatment times. However, in recent years, extracellular vesicles (EVs) have gained significant attention due to their exceptional osteogenic function in bone regeneration and superior therapeutic effects compared to traditional cell therapy. EVs have emerged as a promising therapy for tissue defect regeneration due to their various physiological functions, such as regulating the immune response and promoting tissue repair and regeneration. Moreover, EVs have good biocompatibility, low immunogenicity, and long-term stability, and can be improved through pretreatment and other methods. Studies investigating the mechanisms by which extracellular vesicles promote bone regeneration and applying EVs from different sources using various methods to animal models of bone defects have increased. Therefore, this paper reviews the types of EVs used for bone regeneration, their sources, roles, delivery pathways, scaffold biomaterials, and applications.

Research on the model of mandibular alveolar bone defect in rabbits

Objectives: The purpose of this study was to create an animal model of a mandibular alveolar bone defect without compromising the animal’s well-being. Materials and methods: A total of 24 New Zealand white rabbits underwent surgery to create mandibular alveolar bone defects. The animals were sacrificed at 2, 4, 6, 8, 10, and 12 weeks post-surgery. To assess bone regeneration at the surgical site, radiography, dental cone-beam computed tomography (CT), and histological examination using Hematoxylin and Eosin staining were performed on the skull. Results: A straightforward and easily executable method was devised to create the rabbit mandibular alveolar defect model. After 10 weeks, complete soft tissue and bone regeneration were observed. X-ray and cone-beam CT evaluations demonstrated a progressive increase in bone density from weeks 2 to 12. Histological examination revealed that the alveolar bone structure was formed incrementally at the surgical site. The bone and connective tissue had filled the defect after 8 weeks. Conclusion: The creation of a model of mandibular alveolar bone defects in rabbits is a straightforward process that can be used to assess the regeneration of alveolar bone at the defect site. This animal model can serve as the foundation for tests to evaluate the capacity of biomaterials to regenerate the alveolar bone Key words: mandibular alveolar bone, Alveolar bone defects, animal models, bone regenerative medicine, tissue engineering.

MicroRNA-18a regulates the Pyroptosis, Apoptosis, and Necroptosis (PANoptosis) of osteoblasts induced by tumor necrosis factor-α via hypoxia-inducible factor-1α

BackgroundTumor necrosis factor-α (TNF-α) is involved in inflammatory responses and promotes cell death and the inhibition of osteogenic differentiation. MicroRNA (miRNA) plays a crucial role in the infected bone diseases, however, the biological role of miRNAs in inflammation-induced impaired osteogenic differentiation remains unclear. This study aimed to explore the role of miRNA-18a-5p (miR-18a) in regulating PANoptosis and osteogenic differentiation in an inflammatory environment via hypoxia-inducible factor-1α (HIF1-α). MethodsThe expression of miR-18a in MC3T3-E1 cells was analyzed using quantitative reverse transcription-polymerase chain reaction in an inflammatory environment induced by TNF-α. The expression of HIF1-α and NLRP3 in LV-miR-18a or sh-miR-18a cells was analyzed using western blotting. Fluorescence imaging for cell death, flow cytometry, and alkaline phosphatase activity analysis were used to analyze the role of miR-18a in TNF-α-induced PANoptosis and the inhibition of osteogenic differentiation. An animal model of infectious bone defect was established to validate the regulatory role of miR-18a in an inflammatory environment. ResultsThe expression of miRNA-18a in the MC3T3-E1 cell line was significantly lower under TNF-α stimulation than in the normal environment. miR-18a significantly inhibited the expression of HIF1-α and NLRP3, and inhibition of HIF1-α expression further inhibited NLRP3 expression. Furthermore, inhibition of miR-18a expression promoted the TNF-α-induced PANoptosis and inhibition of osteogenic differentiation, whereas miR-18a overexpression and the inhibition of both HIF1-α and NLRP3 reduced the effects of TNF-α. These findings are consistent with those of the animal experiments. ConclusionmiRNA-18a negatively affects HIF1-α/NLRP3 expression, inhibits inflammation-induced PANoptosis, and impairs osteogenic differentiation. Thus, it is a potential therapeutic candidate for developing anti-inflammatory strategies for infected bone diseases.

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.

Static magnetic field enhances the bone remodelling capacity of human demineralized bone matrix in a rat animal model of cranial bone defects.

The regeneration of bone defects that exceed 2 cm is a challenge for the human body, necessitating interventional therapies. Demineralized bone matrices (DBM) derived from biological tissues have been employed for bone regeneration and possess notable osteoinductive and osteoconductive characteristics. Nevertheless, their efficiency in regenerating critically sized injuries is limited, and therefore additional signaling cues are required. Thanks to the piezoelectric properties of the bone, external physical stimulation is shown to accelerate tissue healing. We have implanted human DBM in critically sized cranial bone defects in rat animal models and exposed them to an external magnetic field (1 T) to enhance endogenous bone formation. Our in vitro experiments showed the superior cytocompatibility of DBM compared to cell culture plates. Furthermore, alkaline phosphatase activity after 14 days and Alizarin red staining at 28 days demonstrated differentiation of rat bone marrow mesenchymal stem cells into bone lineage on DBM. Computer tomography images together with histological analyses showed that implanting DBM in the injured rats significantly enhanced bone regeneration. Notably, combining DBM transplantation with a 2 h daily exposure to a 1 T magnetic field for 2 weeks (day 7 to 21 post-surgery) significantly improved bone regeneration compared to DBM transplantation alone. This research indicates that utilizing external magnetic stimulation significantly enhances the potential of bone allografts to regenerate critically sized bone defects.

Ln vitro assessment of immunomodulatory and osteogenic properties in 3D-printed hydroxyapatite/barium titanate piezoelectric ceramic scaffolds

Research has demonstrated a significant correlation between the skeletal and immune systems. Moreover, piezoelectric materials can affect macrophage polarization, thereby enhancing osteogenic differentiation in stem cells. This study involved the preparation of a piezoelectric ceramic scaffold, comprising hydroxyapatite (HA) and barium titanate (BT), using digital light processing (DLP) 3D printing technology. The scaffold's mechanical (compressive strength: 145.5 MPa) and piezoelectric (d33: 2.15 pC N−1) properties closely resemble those of cortical bone. In vitro analyses revealed that the polarized HA/BT ceramic samples are biologically safe and can induce macrophage polarization towards the M2 phenotype (anti-inflammatory), as well as enhance osteogenic differentiation in dental pulp stem cells (DPSCs) when stimulated with low-intensity pulsed ultrasound (LIPUS). In summary, our findings suggest that 3D printed HA/BT piezoelectric ceramic scaffolds, when stimulated by LIPUS, hold significant potential for application in bone tissue engineering, and further research is needed to investigate the performance in bone defect animal models.

Collagen type I-based recombinant peptide promotes bone regeneration in rat critical-size calvarial defects by enhancing osteoclast activity at late stages of healing

IntroductionWe recently demonstrated the bone-forming potential of medium-cross-linked recombinant collagen peptide (mRCP) in animal models of bone defects. However, these studies were limited to a 4-week observation period; therefore, in the present study, we aimed to further evaluate mRCP as a suitable bone graft material for the alveolar cleft by analyzing its bone-forming potential, osteogenic-inducing ability, and biodegradation over an extended period of 12 weeks, using a rat critical-size calvarial defect model. MethodsUsing Sprague-Dawley rats, we created critical-size calvarial defects through a surgical procedure. The defects were then filled with 3 mg of mRCP (mRCP group) or 18 mg of Cytrans® (CA) granules, which has a carbonate apatite-based composition resembling natural bone, was used as a reference material (CA group). For negative control, the defects were left untreated. Bone volume, total bone volume (bone volume including CA granules), and bone mineral density (BMD) in the defect were assessed using micro-computed tomography (μ-CT) at 0, 4, 8, and 12 weeks after implantation. Using histomorphometric analyses of hematoxylin and eosin (H&E)-stained sections, we measured the amount of newly formed bone and total newly formed bone (new bone including CA granules) in the entire defect site, as well as the amount of newly formed bone in the central side, two peripheral sides (left and right), periosteal (top) side, and dura mater (bottom) side. In addition, we measured the amount of residual bone graft material in the defect. Osteoclasts and osteoblasts in the newly formed bone were detected using tartrate-resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP) staining, respectively. ResultsBone volume in the mRCP group increased over time and was significantly larger at 8 and 12 weeks after surgery than at 4 weeks. The bone volume in the mRCP group was greater than that of the CA and control groups at 4, 8, and 12 weeks after implantation, and while the total bone volume was greater in the CA group after 4 and 8 weeks, the mRCP group had comparable levels of total bone volume to that of the CA group at 12 weeks after implantation. The BMD of the mRCP group reached similar levels to native calvaria bone at the same time point. H&E-stained sections revealed a larger amount of newly formed bone 12 weeks after implantation in the mRCP group compared to that of the CA and control groups. The total newly formed bone at 12 weeks after implantation was on par with that in the CA group. Furthermore, at the defect site, the area of newly formed bone was larger on the peripheral and dura mater sides. Notably, the number of osteoclasts in the mRCP group was higher than in the CA and control groups and peaked 8 weeks after implantation, which coincided with the timing of the greatest resorption of mRCP. Although the ALP-positive area was greater in the mRCP group compared to other groups, we did not detect any significant changes in the number of osteoblasts over time. ConclusionThis study demonstrated the bone-forming potential of mRCP over an extended period of 12 weeks, suggesting that mRCP sufficiently resists resorption to promote bone formation through induction of osteoclast activation in the late stages of the healing period.

Fused Deposition Modeling Printed PLA/Nano β-TCP Composite Bone Tissue Engineering Scaffolds for Promoting Osteogenic Induction Function.

Large bone defects caused by congenital defects, infections, degenerative diseases, trauma, and tumors often require personalized shapes and rapid reconstruction of the bone tissue. Three-dimensional (3D)-printed bone tissue engineering scaffolds exhibit promising application potential. Fused deposition modeling (FDM) technology can flexibly select and prepare printed biomaterials and design and fabricate bionic microstructures to promote personalized large bone defect repair. FDM-3D printing technology was used to prepare polylactic acid (PLA)/nano β-tricalcium phosphate (TCP) composite bone tissue engineering scaffolds in this study. The ability of the bone-tissue-engineered scaffold to repair bone defects was evaluated in vivo and in vitro. PLA/nano-TCP composite bone tissue engineering scaffolds were prepared using FDM-3D printing technology. The characterization data of the scaffolds were obtained using relevant detection methods. The physical and chemical properties, biocompatibility, and in vitro osteogenic capacity of the scaffolds were investigated, and their bone repair capacity was evaluated using an in vivo animal model of rabbit femur bone defects. The FDM-printed PLA/nano β-TCP composite scaffolds exhibited good personalized porosity and shape, and their osteogenic ability, biocompatibility, and bone repair ability in vivo were superior to those of pure PLA. The merits of biodegradable PLA and bioactive nano β-TCP ceramics were combined to improve the overall biological performance of the composites. The FDM-printed PLA/nano-β-TCP composite scaffold with a ratio of 7:3 exhibited good personalized porosity and shape, as well as good osteogenic ability, biocompatibility, and bone repair ability. This study provides a promising strategy for treating large bone defects.

Low-temperature deposition 3D printing biotin-doped PLGA/β-TCP scaffold for repair of bone defects in osteonecrosis of femoral head

The removal of necrotic bone and implantation of bone repair materials is an effective treatment for osteonecrosis of the femoral head (ONFH)-type bone defects, but there are currently no clinically applicable bone repair materials. In this study, a biotin-doped bone repair scaffold was created using low-temperature deposition (LTD) three-dimensional (3D) printing technology, and its ability to repair bone defects in ONFH was evaluated. The scaffold was characterized in vitro, and its cytotoxicity and osteogenic capacity were assessed by co-culturing the scaffold with rat bone marrow mesenchymal stem cells. The scaffolds were implanted in an animal model of ONFH-type bone defects, and the effect of scaffolds on promoting bone repair was evaluated by means of radiology and histopathology. LTD 3D-printed biotin-doped scaffolds showed cancellous bone-like structures without inducing cytotoxicity, whereas high-biotin β-TCP scaffolds (HBPT; containing 2% biotin) promoted osteogenic differentiation more effectively. Experiments on animals revealed that the effect of HBPT on bone repair was significantly superior to that of other groups. The in vivo biocompatibility of HBPT was confirmed by blood analysis and hematoxylin and eosin staining of the main organs. In conclusion, biotin-doped scaffolds can be used to treat ONFH-type bone defects by virtue of their ability in promoting bone regeneration.

The role of lactoferrin in bone remodeling: evaluation of its potential in targeted delivery and treatment of metabolic bone diseases and orthopedic conditions.

Lactoferrin (Lf) is a multifunctional protein that is synthesized endogenously and has various biological roles including immunological regulation, antibacterial, antiviral, and anticancer properties. Recently, research has uncovered Lf's critical functions in bone remodeling, where it regulates the function of osteoblasts, chondrocytes, osteoclasts, and mesenchymal stem cells. The signaling pathways involved in Lf's signaling in osteoblasts include (low density lipoprotein receptor-related protein - 1 (LRP-1), transforming growth factor β (TGF-β), and insulin-like growth factor - 1 (IGF-1), which activate downstream pathways such as ERK, PI3K/Akt, and NF-κB. These pathways collectively stimulate osteoblast proliferation, differentiation, and mineralization while inhibiting osteoclast differentiation and activity. Additionally, Lf's inhibitory effect on nuclear factor kappa B (NF-κB) suppresses the formation and activity of osteoclasts directly. Lf also promotes chondroprogenitor proliferation and differentiation to chondrocytes by activating the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and phosphoinositide 3-kinase/protein kinase B(PI3K/Akt)signaling pathways while inhibiting the expression of matrix-degrading enzymes through the suppression of the NF-κB pathway. Lf's ability to stimulate osteoblast and chondrocyte activity and inhibit osteoclast function accelerates fracture repair, as demonstrated by its effectiveness in animal models of critical-sized long bone defects. Moreover, studies have indicated that Lf can rescue dysregulated bone remodeling in osteoporotic conditions by stimulating bone formation and suppressing bone resorption. These beneficial effects of Lf on bone health have led to its exploration in nutraceutical and pharmaceutical applications. However, due to the large size of Lf, small bioactive peptides are preferred for pharmaceutical applications. These peptides have been shown to promote bone fracture repair and reverse osteoporosis in animal studies, indicating their potential as therapeutic agents for bone-related diseases. Nonetheless, the active concentration of Lf in serum may not be sufficient at the site requiring bone regeneration, necessitating the development of various delivery strategies to enhance Lf's bioavailability and target its active concentration to the site requiring bone regeneration. This review provides a critical discussion of the issues mentioned above, providing insight into the roles of Lf in bone remodeling and the potential use of Lf as a therapeutic target for bone disorders.

Injectable and Cell-Laden Hydrogel in the Contained Bone Defect Animal Model: A Systematic Review.

Due to its high water content and biomimetic properties simulating extracellular matrix (ECM), hydrogels have been used as preferred cell culture and delivery systems. Similarly, cell-loaded hydrogels can be easily injected into target areas in a minimally invasive manner, minimizing surgical trauma, adapting to irregular shaped defects, and benefiting patients. In this study, we systematically reviewed multiple studies on hydrogel-based bone defect research and briefly summarized the progress of injectable and cell-loaded hydrogels in bone defect repair. A systematic search was conducted in the PubMed and Web of Science databases using selected search terms. Initially, 185 articles were retrieved from the databases. After full-text screening based on inclusion and exclusion criteria, 26 articles were included in this systematic review. Data collected from each study included culture model, seed cell type and origin, cell concentration, scaffold material, scaffold shape, experimental animal and site, bioactive agents, and binding method. This injectable and cell-loaded hydrogel shows certain feasibility in bone tissue engineering applications. Injectable and cell-loaded hydrogels have been widely applied in bone tissue engineering research. The future direction of bone tissue engineering for bone defect treatment involves the use of new hydrogel materials and biochemical stimulation.

The effects of BaTiO3 on the handleability and mechanical strength of the prepared piezoelectric calcium phosphate silicate for bone tissue engineering

Natural bone is a piezoelectric material that can generate electrical signals when subjected to an external force. Although many studies have attempted to develop piezoelectric biomaterials for bone regeneration, post-treatment steps, such as sintering, are always needed. In this study, we prepared an injectable and piezoelectric bone substitute based on nanosized BaTiO3 (nBT)-added calcium phosphate silicate (CPS). The impacts of nBT on the CPS handleability and mechanical strength were characterized, and show that adding nBT could improve the CPS handleability but affect the CPS mechanical strength in a concentration-dependent manner (from 25.3 ± 1.0 MPa for 10BC to 13.5 ± 1.0 MPa for 40BC). In addition, our approach could fabricate a piezoelectric bone substitute with comparable piezoelectricity to the native bone without any post-treatment. The in vitro analyses demonstrated that nBT/CPS was biocompatible and could promote osteoblast differentiation. In conclusion, our results strongly indicate that the injectable formulation based on nBT/CPS can be a promising candidate in bone tissue engineering, and further research is needed to investigate the biomaterial's performance in bone defect animal models.

Regenerative capacity of trophoblast stem cell-derived extracellular vesicles on mesenchymal stem cells

BackgroundHuman mesenchymal stem cells (MSCs) are therapeutic for clinical applications because of their excellent immunomodulatory and multiple lineage differentiation abilities at tissue injury sites. However, insufficient number of cells and lack of regenerative properties during in vitro expansion still limit the clinical applicability of MSC therapies. Here, we demonstrated a preconditioning strategy with trophoblast stem cell-derived extracellular vesicles (TSC-EVs) to boost the proliferation and regenerative capacity of MSCs.MethodsWe employed cell proliferation analyses such as CCK8 and BrdU assays to determine the proliferation-promoting role of TSC-EVs on MSCs. Osteogenic effects of TSC-EVs on MSCs were assessed by alkaline phosphatase (ALP) activity, calcium assays, and calvarial bone defect animal models. For skin regenerative effects, skin wound mice model was exploited to analyze wound-healing rate in this study, as well as immunofluorescence and histological staining evaluates. We also performed the small RNA profiling and RNA-sequencing analyzes to understand the cellular mechanism of TSC-EVs on MSCs.ResultsTSC-EVs significantly promoted MSC proliferation under xeno-free conditions and facilitated the therapeutic effects of MSCs, including osteogenesis, anti-senescence, and wound healing. Transcriptomic analysis also provided evidence that specific microRNAs in TSC-EVs and differentially expressed genes (DEGs) in TSC-EV-treated MSCs showed the possibility of TSC-EVs triggering the regenerative abilities of MSCs with cytokine interaction. Hence, we found that NGF/Akt signaling mediated the regenerative effects of TSC-EVs on MSCs as a particular cellular signaling pathway.ConclusionThe results of this study demonstrated the functional properties of TSC-EVs on MSCs for MSC-based therapeutic applications, suggesting that TSC-EVs may serve as a potential preconditioning source for MSC therapy in the clinical field of regenerative medicine.Graphical abstract

Development and evaluation of local regenerative biomimetic bone-extracellular matrix scaffold loaded with nano-formulated quercetin for orthopedic fractures

The prevalence of bone injuries is greatly increasing each year and the proper healing of fractures without any complications is very challenging. Self-setting calcium phosphate cements (CPCs) have attracted great attention as bioactive synthetic bone substitutes. Quercetin (QT) is a multipurposed drug with reported bone-conserving properties. The loading of QT and QT-phospholipid complex within nanostructured lipid carriers (NLC) was proposed to overcome the poor physical properties of the drug and to introduce the use of bioactive excipients as phospholipids and olive oil. The aim of this work was to formulate a regenerative scaffold loaded with nano-formulated QT for local treatment of orthopedic fractures.For the first time, scaffolds composed of brushite CPC were prepared and loaded with quercetin lipid nano-systems. In vitro tests proved that the addition of lipid nano-systems did not deteriorate the properties of CPC where QT-NLC/CPC showed an adequate setting time, appropriate compressive strength, and porosity. The scanning electron microscope confirmed maintenance of nanoparticles integrity within the cement. Using a rat femur bone defect animal model, the histological results showed that the QT-NLC/CPC had a superior bone healing potential compared to crude unformulated QT/CPC. In conclusion, QT-NLC /CPC are promising lipid nano-composite materials that could enhance bone regeneration.

Acceleration of Bone Fracture Healing through the Use of Bovine Hydroxyapatite or Calcium Lactate Oral and Implant Bovine Hydroxyapatite-Gelatin on Bone Defect Animal Model.

Bone grafts a commonly used therapeutic technique for the reconstruction and facilitation of bone regeneration due to fractures. BHA-GEL (bovine hydroxyapatite-gelatin) pellet implants have been shown to be able accelerate the process of bone repair by looking at the percentage of new bone, and the contact between the composite and bone. Based on these results, a study was conducted by placing BHA-GEL (9:1) pellet implants in rabbit femoral bone defects, accompanied by 500 mg oral supplement of BHA or calcium lactate to determine the effectiveness of addition supplements. The research model used was a burr hole defect model with a diameter of 4.2 mm in the cortical part of the rabbit femur. On the 7th, 14th and 28th days after treatment, a total of 48 New Zealand rabbits were divided into four groups, namely defect (control), implant, implant + oral BHA, and implant + oral calcium lactate. Animal tests were terminated and evaluated based on X-ray radiology results, Hematoxylin-Eosin staining, vascular endothelial growth Factor (VEGF), osteocalcin, and enzyme-linked immunosorbent assay (ELISA) for bone alkaline phosphatase (BALP) and calcium levels. From this research can be concluded that Oral BHA supplementation with BHA-GEL pellet implants showed faster healing of bone defects compared to oral calcium lactate with BHA-GEL pellet implants.

Core-Shell Structured Porous Calcium Phosphate Bioceramic Spheres for Enhanced Bone Regeneration.

Adequate new bone regeneration in bone defects has always been a challenge as it requires excellent and efficient osteogenesis. Calcium phosphate (CaP) bioceramics, including hydroxyapatite (HA) and biphasic calcium phosphates (BCPs), have been extensively used in clinical bone defect filling due to their good osteoinductivity and biodegradability. Here, for the first time, we designed and fabricated two porous CaP bioceramic granules with core-shell structures, named in accordance with their composition as BCP@HA and HA@BCP (core@shell). The spherical shape and the porous structure of these granules were achieved by the calcium alginate gel molding technology combined with a H2O2 foaming process. These granules could be stacked to build a porous structure with a porosity of 65-70% and a micropore size distribution between 150 and 450 μm, which is reported to be good for new bone ingrowth. In vitro experiments confirmed that HA@BCP bioceramic granules could promote the proliferation and osteogenic ability when cocultured with bone marrow mesenchymal stem cells, while inhibiting the differentiation of RAW264.7 cells into osteoclasts. In vivo, 12 weeks of implantation in a critical-sized femoral bone defect animal model showed a higher bone volume fraction and bone mineral density in the HA@BCP group than in the BCP@HA or pure HA or BCP groups. From histological analysis, we discovered that the new bone tissue in the HA@BCP group was invading from the surface to the inside of the granules, and most of the bioceramic phase was replaced by the new bone. A higher degree of vascularization at the defect region repaired by HA@BCP was revealed by 3D microvascular perfusion angiography in terms of a higher vessel volume fraction. The current study demonstrated that the core-shell structured HA@BCP bioceramic granules could be a promising candidate for bone defect repair.

Influence of donor age and comorbidities on transduced human adipose-derived stem cell in vitro osteogenic potential.

Human adipose-derived mesenchymal stem cells (ASCs) transduced with a lentiviral vector system to express bone morphogenetic protein 2 (LV-BMP-2) have been shown to reliably heal bone defects in animal models. However, the influence of donor characteristics such as age, sex, race, and medical co-morbidities on ASC yield, growth and bone regenerative capacity, while critical to the successful clinical translation of stem cell-based therapies, are not well understood. Human ASCs isolated from the infrapatellar fat pads in 122 ASC donors were evaluated for cell growth characteristics; 44 underwent additional analyses to evaluate in vitro osteogenic potential, with and without LV-BMP-2 transduction. We found that while female donors demonstrated significantly higher cell yield and ASC growth rates, age, race, and the presence of co-morbid conditions were not associated with differences in proliferation. Donor demographics or the presence of comorbidities were not associated with differences in in vitro osteogenic potential or stem cell differentiation, except that transduced ASCs from healthy donors produced more BMP-2 at day 2. Overall, donor age, sex, race, and the presence of co-morbid conditions had a limited influence on cell yield, proliferation, self-renewal capacity, and osteogenic potential for non-transduced and transduced (LV-BMP-2) ASCs. These results suggest that ASCs are a promising resource for both autologous and allogeneic cell-based gene therapy applications.

Facile mussel-inspired polymerization to facilitate biomimetic in situ homogeneous mineralization for bone regeneration

As a mussel-inspired polymer, polydopamine (PDA) coating can be used as a simple and universal way to form nano-hydroxyapatite (nHAP) crystals on a variety of substrates. Therefore, PDA is widely used in the preparation of bone repair materials. However, its uneven distribution and easy degradation under strongly alkaline conditions limit its application. Therefore, based on the principle of in situ regeneration, this study combined the freeze-drying technology with the diffusion method of ammonium carbonate to bionics construct porous micro-nano composite scaffolds. Chitosan (CS) and dopamine hydrochloride (DA) were used as the chief components to form a multilayer layered scaffold structure. PDA and nHAP grow in situ on the surface of the freeze-dried scaffolds after ammonium carbonate gas diffusion treatment and Tris buffer immersion. The mild synthesis method effectively avoided the decomposition of PDA, and the gas diffusion method promoted the uniform distribution of PDA. The effect of PDA on nHAP formation was studied by adjusting the amount of DA. The cross-linking reaction between PDA and CS was verified by FTIR and XPS. Surface morphology was observed by SEM. The formation of nHAP was confirmed by XRD. The scaffolds had good porosity, degradation, water absorption, thermal stability and biomimetic mineralization properties. Cell experiments showed that the scaffolds had good biocompatibility and had no toxicity to cells. The construction of bone defect animal model further verified that the scaffolds promoted bone regeneration in the defect site of the body. This study provided a new idea and method for the preparation of bone repair scaffolds.

Anabolic Effects of a Novel Simvastatin Derivative on Treating Rat Bone Defects.

Large bone defects may develop fracture nonunion, leading to disability and psychosocial burdens. Bone grafting with anabolic agents is a good autografting alternative. Simvastatin, as a cholesterol-lowering agent worldwide, is proven to enhance osteogenesis. Considering its dose-dependent adverse effects, we developed a simvastatin derivative, named KMUHC-01, which has bone anabolic capacity and lower cytotoxicity than simvastatin. We hypothesize that KMUHC-01 could help bone formation in bone-defect animal models. We used rat models of critical calvarial and long-bone defects to evaluate the effects of KMUHC-01 and simvastatin on biological changes at the bone defect through histology, immunohistology, and mechanical testing using three-point bending and evaluated the new bone formation microstructure through microcomputed tomography analysis. The newly formed bone microstructure at the calvarial defect site showed a significantly improved trabecular bone volume in the KMUHC-01 1-μM group compared with that in the control and simvastatin groups. The biomechanical study revealed a significantly increased maximal strength in the KMUHC-01 1-μM group compared with that in the control group. KUMHC-01, as a simvastatin derivative, showed a great anabolic effect in promoting bone defect healing. However, further studies will be conducted to prove the bioavailability and bone-forming efficacy of KMUHC-01 via systemic administration.