Porous Biphasic Calcium Phosphate Research Articles

Effects of void defects on the mechanical properties of biphasic calcium phosphate nanoparticles: A molecular dynamics investigation

Porous biphasic calcium phosphate (BCP) ceramics are widely used in bone tissue engineering, and the mechanical properties of BCP implants must be reliable. However, the effects of pore structure (e.g., shape and size) on the mechanical properties are not well understood. In this study, we used molecular dynamics simulations to investigate the influence of pore shape and size on the mechanical behavior of BCP nanoparticles. BCP void models with cylindrical and cuboid pores ranging from 2 to 16 nm in diameter were constructed, and the elastic moduli were calculated. In addition, uniaxial tensile and compressive tests were performed on the models. We found that the pore size had a more significant impact on the mechanical properties of BCP than pore shape. Further, the elastic moduli decreased nonlinearly with increasing pore size. In addition, the tensile and compressive strength also decreased with the increase in pore size, but the ductility improved. Furthermore, deformation and fracture were more likely to occur near the pores and at the phase interfaces as a result of high atomic local strain in the calcium-deficient hydroxyapatite area. The results of this work reveal the effects of pore parameters on the mechanical properties of porous BCP at the nanometer level, which may aid the design of improved porous and multiphase CaP-based biomaterials for bone regeneration.

3D gel-printing of hierarchically porous BCP scaffolds for bone tissue engineering

In this study, a conventional technique of porous preparation was used to improve the constructive capability of direct ink writing on microstructures, and the hierarchically porous scaffolds were successfully prepared by 3D gel printing (3DGP). Micron-sized hydroxyapatite (HA) was coated with tricalcium phosphate (TCP) nanopowders synthesized by chemical co-precipitation to form biphasic calcium phosphate (BCP). The random structure of concave micropores was achieved by filling the BCP slurry with PMMA microspheres while successfully controlling the internal porosity of printed filaments. The results showed that the three-stage porous structure was successfully constructed, i.e., macroscopic pores of 1.50-2.00 mm, spherical micropores of 100-200 μm, and inter-powder interstices of 1.00-10.00 μm. Nano-TCP coated micron-HA powders improved the sintering activity of BCP particles. The compressive strength and porosity of the scaffolds sintered at 1400 °C were 2.78 MPa and 84.98%. The hierarchically porous BCP scaffolds had bright applications in bone tissue engineering.

Preparation and evaluation of osteoinductive porous biphasic calcium phosphate granules obtained from eggshell for bone tissue engineering

The synthetic bone graft substitutes currently used clinically are osteoconductive but not osteoinductive; their low success rate is thus their biggest disadvantage. The use of biomass raw materials for synthesizing calcium phosphate has gradually attracted increased research attention. In this study, hydroxyapatite powder was prepared through liquid-phase precipitation from eggshell, and porous biphasic calcium phosphate granules (EBGs) were then obtained using a pore former and sintering procedure. The EBGs were discovered to have high biocompatibility and no cytotoxicity. The results of animal experiments showed that the area of new bone growth was high, numerous Haversiancanals could be observed, and almost all EBGs were surrounded by new bone tissue, which proved that the EBGs had excellent osteoinductivity. By contrast, numerous fat cells were found in the femoral defect area when a commercial bone graft was employed. Various biological inorganic ions (Mg, Sr, Na, and Fe) originally in the eggshell raw materials were incorporated into the EBGs, and the EBGs exhibited excellent osteogenic abilities. The developed approach provides an economical and feasible solution for the treatment of bone defects.

Microstructures and mechanical properties of biphasic calcium phosphate bioceramics fabricated by SLA 3D printing

Porous biphasic calcium phosphate (BCP) bioceramics are considered to be the most promising bone repair materials in clinical medicine. Stereolithography (SLA) 3D printing can precisely fabricate bioceramic scaffolds with complex porous structure. During this process, how to obtain a suitable sintering procedure for BCP bioceramics by SLA 3D printing is the key to determine the microstructures and mechanical properties, but this is absent. In this present study, we prepared BCP bioceramics with superior densification and mechanical properties by SLA 3D printing, and mainly investigated the effects of sintering temperature and condition on the microstructures and mechanical properties of SLA 3D printed BCP bioceramics for the first time. At the optimized procedure of sintering temperature 1250 °C for 2 h, the 3D printed BCP bioceramics showed uniform shrinkage in all directions, and especially the mechanical properties were close to that of human cortical bone. Furthermore, complex porous BCP scaffolds with high porosity (51.49 %) and compressive strength (8.14 MPa) were successfully obtained. BCP bioceramics greatly promoted the proliferation of MC3T3-E1 cells by the release of Ca and P ion and presented excellent bioactivity. This work proves that SLA 3D printing technology can prepare BCP bioceramics with high mechanical and functional properties, and provides a new route for manufacturing high performance BCP bioceramic scaffolds with complex structure by SLA 3D printing for repairing bone defect in clinical.

Preparation and characterization of two novel osteoinductive fishbone-derived biphasic calcium phosphate bone graft substitutes.

Many studies have reported on the conversion of natural resources into xenografts with hydroxyapatite (HA) as major component, but the extraction of biphasic calcium phosphate (HA/β-TCP) from animal bones and transformation into bone graft substitutes are rarely reported. In this research, two kinds of fish bones were made into granular porous biphasic calcium phosphate bone graft substitutes with particle sizes between 500 to 1000μm through a series of preparation procedures (Salmo salar calcined at 900°C named Sa900 and Anoplopoma fimbria calcined at 800°C named An800). The chemical composition was characterized by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The morphology and porous structure of the scaffolds were comparatively analyzed by scanning electron microscopy (SEM) and mercury porosimeter. The specific surface area of materials was measured by the nitrogen adsorption technique based on BET theory. Cytotoxicity and ectopic osteogenesis were also carried out to investigate the biocompatibility and osteoinductive potential of these materials. The results showed that both fishbone-derived scaffolds were composed of HA and β-TCP with different proportions, and numerous interconnected pores with different sizes were observed at the surface of materials. An800 had higher total porosity reaching 74.8% with higher interconnectivity and micropores mostly distributed at 0.27μm and 0.12μm, while Sa900 had a higher specific surface area and higher intraparticle porosity with nanopores mostly distributed at 0.07μm. CCK-8 assays and Live/dead staining demonstrated excellent biocompatibility. Material-induced osteoid formation were observed on the interface of both internal pores and periphery of materials after implantation in muscle pouch of Wistar rats for 8weeks which indicated some extent of osteoinductive potential of materials. The possible mechanism of material-induced osteogenesis and the effects of chemical composition, surface topography, and spatial structure on osteogenesis were also discussed in this paper.

Biological evaluation of polydopamine and chitosan composite coatings on the 3D printed porous biphasic calcium phosphate scaffold

A current, gradually growing trend is to use bioactive ceramics to develop new bone scaffolds to replace traditional bone implants. Biphasic calcium phosphate (BCP), a mixed material of hydroxyapatite and tricalcium phosphate, has limited osseointegration ability. Therefore, improving the bioactivity and osteoinductive ability of BCP scaffolds through various surface modification methods is an important issue. Polydopamine (PDA) coating, chitosan (CS) coating, and their composite coatings have been previously reported to individually possess a good ability to improve scaffold bioactivity, but few comparisons between them and the optimal coating sequence of PDA and CS have been reported. Herein, we fabricated a BCP scaffold by using a self-made 3D printer, sintered the scaffold, and then prepared PDA, CS, PDA/CS and CS/PDA coatings by solution immersion. Afterwards, the surface morphology, element composition and proportion, compressive strength and surface contact angle of the five groups of scaffolds were characterized. Finally, the adhesion, proliferation and osteogenesis-related gene expression levels of MC3T3-E1 on the five groups in vitro were evaluated in detail. The results showed that the five types of scaffolds were fabricated successfully and that the PDA/CS and CS/PDA coatings had different surface morphologies. The compressive strengths of the composite-coating groups were higher than those of the single-coating groups and the BCP group. The CCK-8 experiment revealed that the composite coatings can promote the initial adhesion and proliferation of cells. The detection results of three osteogenesis-related genes (OCN, COL-1 and Runx-2) proved that the composite coatings had a synergistic effect in promoting osteodifferentiation and that the CS/PDA coating had the greatest effect in improving cell viability and promoting osteogenic differentiation.

Digital light processing (DLP) of nano biphasic calcium phosphate bioceramic for making bone tissue engineering scaffolds

Nano-sized ceramic particles are beneficial for the properties of 3D printed structures via digital light processing (DLP). However, achieving optimal conditions for the preparation and processing of ceramic nanoparticle-based slurry is a significant challenge. In this work, a stable and well dispersed slurry with nano-sized biphasic calcium phosphate (BCP) powders was developed and porous BCP bioceramic scaffolds with good bioactivity were accurately fabricated via the DLP additive manufacturing technology. Effects of dispersant, photoinitiator and solid loading on rheological properties and curing abilities of BCP slurries were systematically studied. The slurry having 65 wt % of nano-sized BCP powders showed a relatively low viscosity of 400 mPa s at the 50 s−1 shear rate. It was found that the optimal photoinitiator concentration was tightly associated with the energy dosage in DLP. Furthermore, the effects of sintering temperature (1100 °C, 1200 °C and 1300 °C) on the surface morphology, shrinkage, phase constitution, hardness, compressive properties, and in vitro biological performance of DLP-formed bioceramics were comprehensively investigated. The results showed that DLP-formed BCP ceramics sintered at 1300 °C had the best mechanical properties, in vitro cytocompatibility, and homogeneous bone-like apatite formation ability. Finally, DLP-formed 3D scaffolds with different pore sizes ranging from 300 μm to 1 mm were successfully fabricated, which exhibited high fidelity and accuracy. The current study has demonstrated the great potential of using DLP technology to construct functional complex BCP bioceramic scaffolds for bone tissue regeneration.

Porous Biphasic Calcium Phosphate Granules from Oyster Shell Promote the Differentiation of Induced Pluripotent Stem Cells.

Oyster shells are rich in calcium, and thus, the potential use of waste shells is in the production of calcium phosphate (CaP) minerals for osteopathic biomedical applications, such as scaffolds for bone regeneration. Implanted scaffolds should stimulate the differentiation of induced pluripotent stem cells (iPSCs) into osteoblasts. In this study, oyster shells were used to produce nano-grade hydroxyapatite (HA) powder by the liquid-phase precipitation. Then, biphasic CaP (BCP) bioceramics with two different phase ratios were obtained by the foaming of HA nanopowders and sintering by two different two-stage heat treatment processes. The different sintering conditions yielded differences in structure and morphology of the BCPs, as determined by scanning electron microscopy (SEM), X-ray diffraction (XRD), and Brunauer–Emmett–Teller (BET) surface area analysis. We then set out to determine which of these materials were most biocompatible, by co-culturing with iPSCs and examining the gene expression in molecular pathways involved in self-renewal and differentiation of iPSCs. We found that sintering for a shorter time at higher temperatures gave higher expression levels of markers for proliferation and (early) differentiation of the osteoblast. The differences in biocompatibility may be related to a more hierarchical pore structure (micropores within macropores) obtained with briefer, high-temperature sintering.

Biodegradable Elastic Sponge from Nanofibrous Biphasic Calcium Phosphate Ceramic as an Advanced Material for Regenerative Medicine

AbstractBiodegradable porous calcium phosphate (CaP) ceramics are widely used as synthetic graft substitutes for bone regeneration, owing to their chemical and structural similarity to bone and associated bioactivity in terms of bone‐bonding, osteoconductive, and even osteoinductive properties. Nevertheless, the intrinsic brittleness and poor processability of porous CaP ceramics strongly impair their clinical applicability. Herein, a biphasic calcium phosphate (BCP) sponge is developed that consists of a self‐supporting network of seamlessly interwoven hydroxyapatite nanowires and β‐tricalcium phosphate nanofibers and possesses a highly interconnected porous structure with open cell geometry and ultrahigh porosity. Owing to its unique properties, the ceramic sponge can be easily processed into various shapes and dimensions, such as cylindrical scaffolds and thin, flexible membranes. Moreover, the BCP sponge can be introduced into a bone defect in a compacted or folded state from a syringe and, upon wetting, expand to its original shape, thereby filling the cavity. The nanofibrous sponge gradually degrades in vitro and rapidly mineralizes when immersed in simulated body fluid. Moreover, it adsorbs significantly more proteins than a conventional porous BCP ceramic. Finally, the nanofibrous sponge supports the attachment, proliferation, and osteogenic differentiation of human mesenchymal stromal cells comparable to the conventional porous BCP ceramic.

Role of Na+, K+-ATPase ion pump in osteoinduction

Porous biphasic calcium phosphate bioceramic (BCP) possesses osteoinductivity to induce the osteoblastic commitment of mesenchymal stem cells (MSCs) and ectopic bone formation. However, the underlying mechanism remains enigmatic. We performed a gene array analysis of MSCs cocultured with BCP to screen for candidate osteoinductive modulators. Na+, K+-ATPase (NKA), an ion transporter, therefore was identified as a crucial ion transporter in regulating the osteogenesis of the cells. NKA activator, a polyclonal antibody, enriched the cytomembrane abundance of NKA and lead to an enhanced osteogenic effect of BCP. As indicated in gene array analysis and suggested by co-immunoprecipitation assay, protein phosphatase 2A (PP2A) was elevated by BCP to dephosphorylate NKA and prevent its endocytosis. The inhibition of NKA by ouabain resulted in an adverse effect on osteoinductivity of BCP. We further altered NKA activity in mice implanted with BCP and found that the intensity and incidence of osteoinduction was increased by the NKA activator. We went one step further by investigating the potential of targeting NKA in osteoporotic bone regeneration. Activating NKA upregulated osteogenic gene expression and calcium deposition ability of osteoporotic osteoblasts. Furthermore, activation of NKA in mice ameliorated estrogen-deficiency induced bone loss, in terms of increased bone mass and improved bending strength. With this osteoinductive bioceramic derived ion transporter target, we demonstrate that the activation of NKA has significant potential to revolutionize the regeneration of bone. Statement of significanceIn this study, we identified an important role of Na+, K+-ATPase (NKA) have played in osteoinductivity of biphasic calcium phosphate bioceramic (BCP). Furthermore, we demonstrated the therapeutic potential of targeting NKA in osteoporotic bone regeneration. Numerous gene and protein targets to treat osteoporosis were discovered every year, mainly obtained by genomic and proteomic screenings of a large population. In contrast, our study identified an unrevealed bone regenerating target from the upregulated genes induced by an osteoinductive biomaterial. The approach was cost-saving since it did not require a large sample pool. Furthermore, the target derived from this approach was proven to be anabolic. Identification of an anabolic agent holds significant value since most of the current anti-osteoporotic therapies are antiresorptive.

Immunohistochemical characteristics of lateral bone augmentation using different biomaterials around chronic peri-implant dehiscence defects: An experimental in vivo study.

To investigate the immunohistochemical characteristics of a highly porous synthetic bone substitute and a cross-linked collagen membrane for guided bone regeneration. Three experimental groups were randomly allocated at chronic peri-implant dehiscence defect in 8 beagle dogs: (i) biphasic calcium phosphate covered by a cross-linked collagen membrane (test group), (ii) deproteinized bovine bone mineral covered by a natural collagen membrane (positive control) and (iii) no treatment (negative control). After 8 and 16weeks of submerged healing, dissected tissue blocks were processed for immunohistochemical analysis. Seven antibodies were used to detect the remaining osteogenic and angiogenic potential, and quantitative immunohistochemical analysis was done by software. The antigen reactivity of alkaline phosphatase was significantly higher in the test group compared to the positive and negative controls, and it maintained till 16weeks. The intensity of osteocalcin was significantly higher in the positive control at 8weeks than the other groups, but significantly decreased at 16weeks and no difference was found between the groups. A significant large number of TRAP-positive cells were observed in the test group mainly around the remaining particles at 16weeks. The angiogenic potential was comparable between the groups showing no difference in the expression of transglutaminase II and vascular endothelial growth factor. Guided bone regeneration combining a highly porous biphasic calcium phosphate synthetic biomaterial with a crosslinked collagen membrane, resulted in extended osteogenic potential when compared to the combination of deproteinized bovine bone mineral and a native collagen membrane.

Dual-scale porous biphasic calcium phosphate gyroid scaffolds using ceramic suspensions containing polymer microsphere porogen for digital light processing

This study demonstrates a novel type of biphasic calcium phosphate (BCP) gyroid scaffolds featuring of gyroid macroporous structure and micropous BCP walls using poly(methyl methacrylate) (PMMA) microspheres as the porogen for ceramic digital light processing (DLP) technique. To tailor the microporosity of the BCP walls and the overall porosity of the dual-scale porous BCP scaffolds, the PMMA content with regard to the BCP powder was controlled in the range of 40 vol% to 70 vol%. After debinding at 600 °C and sintering at 1200 °C for 3 h, micropores were uniformly created throughout each BCP framework, while preserving 3−dimensional gyroid macroporous structures. As the PMMA content increased from 40 vol% to 70 vol%, the microporosity remarkably increased from 31.9 (±2.5) vol% to 55.2 (±1.4) vol%. This approach allowed the achievement of very high overall porosities (82.2–89.7 vol%) for the dual-scale porous scaffolds. However, all the scaffolds showed reasonable compressive strengths (0.8 MPa −2.1 MPa), which are comparable to those of cancellous bones.

Maxillary sinus lift response to platelet-rich plasma associated with autogenous bone, ceramic biphasic HA/\u03b2-TCP (70:30), or deproteinized bovine bone

BackgroundThis study evaluated the long-term effects of platelet-rich plasma (PRP) on bone formation and regeneration when associated with autogenous bone graft (AB), porous biphasic calcium phosphate (pBCP), or deproteinized bovine bone (DBB) in maxillary sinus augmentation (MSA) of rabbit.MethodsIn 54 rabbits, bilateral MSA procedure was performed and randomly one sinus was filled with 200 mm3 material plus blood clot (AB/clot, DBB/clot, and pBCP/clot) and other with the same graft plus PRP (AB/PRP, DBB/PRP, and pBCP/PRP). After 30, 60, and 180 days, microtomographic were performed to analyze the three-dimensional MSA volume and histomorphometric analyses for the percentage of bone and soft tissues ingrowth. Data were compared by two-way ANOVA and the means were compared by the Tukey test, at p < 0.05.ResultsThe percentage of pBCP and DBB were nearly unchanged throughout the whole period and bone formation occurred in the spaces between particles. The MSA volume filled with DBB and pBCP agglutinated with clot and PRP maintained constant during all experimental periods (147.2 mm3 and 154.9 mm3, respectively, p = 0.7377), and no significant changes in the new formatted bone and soft tissue were observed between treatments. In AB/clot and AB/PRP, the MSA volume was similar at 30 days (140.3 mm3 and 137.9 mm3, respectively), but a higher and gradual reduction was observed until 180 days. In the AB/PRP, this reduction was significantly higher (44.2%) than AB/clot (22.5%) (p = 0.01792). Histologically, the addition of PRP to AB accelerated the new bone formation/remodeling maintaining the percentage of new bone similar to AB/clot during all experimental volume (p = 0.6406), while the AB particles showed a higher resorption in AB/PRP than AB/clot until 60 days (mean of 7.8% and 15.1%, respectively, p = 0.0396).ConclusionThe association of PRP with the autogenous graft accelerates the process of bone formation/remodeling in MSA, but not had influence on the pBCP and DBB groups.

Concentration-dependent effects of latex F1-protein fraction incorporated into deproteinized bovine bone and biphasic calcium phosphate on the repair of critical-size bone defects.

F1-protein fraction (F1) is a natural bioactive compound extracted from the rubber tree, Hevea brasiliensis, and has been recently studied for its therapeutic potential in wound healing. In this study, we investigated the concentration-dependent effects of F1 (0.01%, 0.025%, 0.05%, and 0.1%) incorporated into deproteinized bovine bone (DBB) and porous biphasic calcium phosphate (pBCP), on the repair of rat calvarial critical-size bone defects (CSBD). The defects were analyzed by 3D-microtomography and 2D-histomorphometry at 12 weeks postsurgery. The binding efficiency of F1 to pBCP (96.3 ± 1.4%) was higher than that to DBB (67.7 ± 3.3%). In vivo analysis showed a higher bone volume (BV) gain in all defects treated with DBB (except in 0.1% of F1) and pBCP (except in 0.05% and 0.1% of F1) compared to the CSBD without treatment/control group (9.96 ± 2.8 mm3 ). DBB plus 0.025% F1 promoted the highest BV gain (29.7 ± 2.2 mm3 , p < .0001) compared to DBB without F1 and DBB plus 0.01% and 0.1% of F1. In the pBCP group, incorporation of F1 did not promote bone gain when compared to pBCP without F1 (15.9 ± 4.2 mm3 , p > .05). Additionally, a small BV occurred in defects treated with pBCP plus 0.1% F1 (10.4 ± 1.4 mm3, p < .05). In conclusion, F1 showed a higher bone formation potential in combination with DBB than with pBCP, in a concentration-dependent manner. Incorporation of 0.25% F1 into DBB showed the best results with respect to bone formation/repair in CSBD. These results suggest that DBB plus 0.25% F1 can be used as a promising bioactive material for application in bone tissue engineering.

The use of biphasic calcium phosphate with or without bone morphogenic protein-2 for bone regeneration during immediate implant placement in a micropig model: A pilot study

PurposeThe aim of this pilot study is to investigate bone regeneration in a three wall defect during immediate implant placement, using porous biphasic calcium phosphate (BCP) granules with or without recombinant human bone morphogenetic protein – 2 (rhBMP-2). Materials and MethodsAlveolar defects were created in the mandible of five female, adult micropigs. The lower right 4th premolar was extracted and the buccal cortex removed. Immediate implants were placed and the defect was grafted. Control group (n = 2), received BCP granules and fibrin sealant. The test group (n = 3) received BCP granules in combination with fibrin sealant and rhBMP-2. A collagen membrane was placed over the graft and primary closure of the wound was achieved. The pigs were all euthanized at 6 months after the first surgery. Histology evaluation and histomorphometry analyses calculating % bone implant contact (%BIC) and % bone area (%BA) were performed. ResultsOne pig in the BMP group died prematurely at 3 months after the first surgery. At specimen harvest, there were no dehiscence of all surgical sites and implant survival rate was 100%. The control group demonstrated no new bone formation at the buccal surface of the dental implants at 6 months. The mean %BIC for the BMP group at 6 months was 75.53 ± 12.52%. The mean %BA for the BMP group 6 months was 52.82 ± 3.27%. ConclusionWithin the limitations of this pilot study, the use of rhBMP-2 resulted in significant new bone formation in a non-contained defect around an immediate implant.

Porous calcium phosphate-collagen composite microspheres for effective growth factor delivery and bone tissue regeneration.

Microspheres are beneficial for filling defects of various shapes and provide a large surface area for cell attachment. Porous microspheres have attracted particular attention because they can deliver cells and bioactive molecules such as growth factors. In this study, BCP-collagen composite microspheres were developed for growth factor delivery in bone regeneration. Firstly, porous biphasic calcium phosphate (BCP) microspheres were fabricated by applying a water-in-oil emulsion technique using camphene as a pore generator. Then, porous BCP-collagen composite microspheres were fabricated by repetitively dip coating the microspheres in a collagen solution to effectively deliver growth factor to bone defects. Characterization of the microspheres and in vitro studies were conducted to investigate the effect of collagen infiltration on bone regeneration. In addition, in vitro evaluation demonstrated the sustained bone morphogenetic protein-2 (BMP-2) delivery of the microspheres and the effect of cell differentiation, and in vivo assessment with rabbits revealed that the microspheres filled the defect well and that bone could be regenerated through the microspheres. Moreover, the composite system was more effective for bone regeneration than the bare BCP microspheres because of the drug retention of collagen. These findings indicate that the porous microspheres are effective for tissue regeneration by continuous growth factor delivery.

Fabrication and preliminary evaluation of the osteogenic potential for micro-/nano-structured porous BCP ceramics

Nanoscale interface of biomaterials exhibits superiorities in promoting tissue repair and healing. Bioceramics are one of the most commonly used biomaterials for biological purposes such as tissue engineering. The fabrication of nanoscale ceramics with smart topography is essential for modulating cells in contact with such surface. In this research, a gas foaming method was employed to fabricate porous biphasic calcium phosphate (BCP) ceramics as substrates with a controllable component of hydroxyapatite/beta tricalcium phosphate (HA/β-TCP). Hydrothermal reaction was then introduced to modify the surface morphology of the substrates to obtain a nanoscale topography at different pH values and ionic microenvironments. Three kinds of ceramics (BCP; BCP with a nanoneedle-like surface, BNN; BCP covered with a hollow nanorod-like surface, BHN) were prepared with different surface morphologies and roughness, degradation properties and the phase composition during the hydrothermal process. All of them possessed an interconnected pore structure of macropores and micropores. Importantly, the nano-scale surface structure increased the total pore volumes as well as the micropore volumes, thus enhanced the specific surface area (SSA) and subsequent biological performance. Their biological behavior has also been assessed through a series of pivotal events, i.e. protein adsorption, cell attachment, proliferation, and osteogenic differentiation. Consequently, we found the nano-crystallization of the hollow nanorod-like structured and nanoneedle-like structured ceramic surface enhance the adsorption of both total serum proteins and BSA in vitro compared to the original BCP ceramics with micro-sized grains, as well as exhibit a more persistent release of BSA. Additionally, all the three ceramics have displayed excellent biocompatibilities for MSCs, and the morphology of ceramics played a decisive role in cells spreading and morphology, in which BNN showed the strongest effect to facilitate MSCs differentiation towards osteogenesis.

Osteoinductivity of Porous Biphasic Calcium Phosphate Ceramic Spheres with Nanocrystalline and Their Efficacy in Guiding Bone Regeneration.

Conventional biphasic calcium phosphate (BCP) bioceramics are facing many challenges to meet the demands of regenerative medicine, and their biological properties are limited to a large extent due to the large grain size in comparison with nanocrystalline of natural bone mineral. Herein, this study aimed to fabricate porous BCP ceramic spheres with nanocrystalline (BCP-N) by combining alginate gelatinizing with microwave hybrid sintering methods and investigated their in vitro and in vivo combinational osteogenesis potential. For comparison, spherical BCP granules with microcrystalline (BCP-G) and commercially irregular BCP granules (BAM, BCP-I) were selected as control. The obtained BCP-N with specific nanotopography could well initiate and regulate in vitro biological response, such as degradation, protein adsorption, bone-like apatite formation, cell behaviors, and osteogenic differentiation. In vivo canine intramuscular implantation and rabbit mandible critical-sized bone defect repair further confirmed that nanotopography in BCP-N might be responsible for the stronger osteoinductivity and bone regenerative ability than BCP-G and BCP-I. Collectedly, due to nanotopographic similarities with nature bone apatite, BCP-N has excellent efficacy in guiding bone regeneration and holds great potential to become a potential alternative to standard bone grafts in bone defect filling applications.

Effects of Combined Magnetic Fields Treatment and Nano-Hydroxyapatite Coating on Porous Biphasic Calcium Phosphate Bone Graft in Rabbit Spinal Fusion Model.

An animal experimental study was designed to investigate the efficacy of combined magnetic fields (CMF) treatment and nano-hydroxyapatite (HA) coating in the biphasic calcium phosphate (BCP) graft in posterolateral lumbar fusion. To evaluate the effects of CMF treatment and nano-HA/BCP and their combination effect in posterolateral lumbar fusion. Enhancement of artificial bone graft bioeffects could improve spinal fusion outcomes. The bone graft integration is vital in spinal fusion, nano-HA coating, and CMF treatment were reported as effective methods to improve bone graft integration. A bilateral transverse process fusion model was performed on 32 rabbits. The CMF treatment was performed for 30 minutes per day postoperation. The fusion rate, new bone formation, artificial bone graft-autologous bone fusion interface in x-ray and scanning electron microscopy, biomechanics property of fusion rate, histological fusion condition, artificial bone residual rate, and immunohistochemistry assessment of bone morphogenetic protein 2 (BMP-2) and Transforming growth factor beta 1 (TGF-β1) expression were observed at 9th week after surgery. CMF treatment and nano-HA coating increased the fusion rate, adjusted optical density index, intensity of binding of artificial and autologous bone, bone growth rate, and bending stiffness. CMF treatment also significantly increased BMP-2 and TGF-β1 expression in fusion region while nano-HA coating significantly decreased artificial bone residual rate. Our findings suggest that porous nano-HA/BCP graft could significantly improve spine fusion outcome with excellent bioactivity, biocompatibility and degradability and CMF treatment could significantly improve spine fusion outcome by improving bioactivity and biocompatibility of artificial bone graft in rabbit. Combination of CMF treatment with nano-HA/BCP graft could significantly increase posterolateral lumbar fusion rate in rabbit, which would be a potential strategy for spine fusion preclinical study. N/A.

Enhanced osteoinductivity of porous biphasic calcium phosphate ceramic beads with high content of strontium-incorporated calcium-deficient hydroxyapatite.

Further biomimicking natural bone and enhancing osteoinductivity to meet the requirements of regenerative medicine is the key development direction of biphasic calcium phosphate (BCP) ceramics. Bone mineral is a kind of Ca-deficient hydroxyapatite (CDHA) with many kinds of trace ions incorporated; however, little is actually known regarding the incorporation of trace ions in CDHA, and trace-ions-incorporated CDHA in BCP ceramics has seldom been studied. The present study introduces an effective approach to fabricate porous BCP ceramic beads with a high content of strontium (Sr)-incorporated CDHA (BCP-Sr), and investigated its biological performance, especially with regard to osteoinductivity. The obtained BCP-Sr possessed a good spherical shape, interconnected pore structure, and a high content of Sr-incorporated CDHA phase. Compared to the commercial BCP ceramic irregular granules (BAM® P2040, BCP-C), BCP-Sr had more micropores, relatively faster degradation, better bone-like apatite formation, higher protein adsorption abilities, and was more likely to promote the related osteogenic genes and protein expressions of BMSCs. Further in vivo canine intramuscular implantation confirmed that BCP-Sr had higher osteoinductivity than BCP-C. Collectively, the enhanced osteoinductivity of porous BCP ceramic beads by introducing a high content of Sr-incorporated CDHA has significant implications for designing highly bioactive BCP ceramics for applying in regenerative medicine.