A Bioactive Degradable Composite Bone Cement Based on Calcium Sulfate and Magnesium Polyphosphate.

Calcium sulfate bone cement (CSC) is extensively used as a bone repair material due to its ability to self-solidify, degradability, and osteogenic ability. However, the fast degradation, low mechanical strength, and insufficient biological activity limit its application. This study used magnesium polyphosphate (MPP) and constructed a composite bone cement composed of calcium sulfate (CS), MPP, tricalcium silicate (C3S), and plasticizer hydroxypropyl methylcellulose (HPMC). The optimized CS/MPP/C3S composite bone cement has a suitable setting time of approximately 15.0 min, a compressive strength of 26.6 MPa, and an injectability of about 93%. The CS/MPP/C3S composite bone cement has excellent biocompatibility and osteogenic capabilities; our results showed that cell proliferation is up to 114% compared with the control after 5 days. After 14 days, the expression levels of osteogenic-related genes, including Runx2, BMP2, OCN, OPN, and COL-1, are about 1.8, 2.8, 2.5, 2.2, and 2.2 times higher than those of the control, respectively, while the alkaline phosphatase activity is about 1.7 times higher. Therefore, the CS/MPP/C3S composite bone cement overcomes the limitations of CSC and has more effective potential in bone repair.

Effects of ATP on the Physicochemical Properties and Cytocompatibility of Calcium Sulfate/Calcium Citrate Composite Cement.

Adenosine triphosphate (ATP), acting as a source of energy, has effects on cellular activities, such as adhesion, proliferation, and differentiation. In this study, ATP-loaded calcium sulfate hemihydrate/calcium citrate tetrahydrate cement (ATP/CSH/CCT) was successfully prepared for the first time. The effect of different contents of ATP on the structure and physicochemical properties of ATP/CSH/CCT was also studied in detail. The results indicated that incorporating ATP into the cement did not significantly alter their structures. However, the addition ratio of ATP directly impacted the mechanical properties and in vitro degradation properties of the composite bone cement. The compressive strength of ATP/CSH/CCT gradually decreased with an increasing ATP content. The degradation rate of ATP/CSH/CCT did not significantly change at low concentrations of ATP, but it increased with a higher ATP content. The composite cement induced the deposition of a Ca-P layer in a phosphate buffer solution (PBS, pH = 7.4). Additionally, the release of ATP from the composite cement was controlled. The ATP was controlled releasing at the 0.5% and 1% ATP in cement by the diffusion of ATP and the degradation of the cement, whereas it was controlled by the diffusion process merely at the 0.1% ATP in cement. Furthermore, ATP/CSH/CCT demonstrated good cytoactivity with the addition of ATP and is expected to be used for the repair and regeneration of bone tissue.

Ni(I)-Catalyzed Hydroxylation of Aryl Halides with Water under Thermal Catalysis.

A highly efficient hydroxylation of (hetero)aryl halides using water as a hydroxyl source via Ni catalysis promoted by PhSiH3 under thermal catalysis is reported. This methodology provides a general procedure to obtain diverse multifunctional pharmaceutically phenols and polyphenols, some of which are proven challenging to be synthesized using literature methods. Mechanism studies demonstrated that the addition of PhSiH3 led to the generation of active Ni(I) species, which catalyze the hydroxylation via a Ni(I)-Ni(III) pathway.

Effect of ZnO-doped magnesium phosphate cements on osteogenic differentiation of mBMSCs in vitro.

The insufficient osteogenesis of magnesium phosphate cements (MPCs) limits its further application. It is significant to develop a bioactive MPC with osteogenic properties. In this work, MPCs were reinforced by zinc oxide nanoparticles (ZnO-NPs). The composition, microstructure, setting time, compressive strength and degradation of ZnO-NPs/MPCs (ZNMPCs) were evaluated. The results showed that the setting times of MPCs were prolonged from 8.2 to 25.3 min (5.0ZNMPC). The exothermic temperatures were reduced from 45.8 ± 0.4℃ (MPCs) to 39.3 ± 0.5℃ (1.0ZNMPC). The compressive strength of ZNMPC composite cement with 1 wt. % ZnO-NPs (1.0ZNMPC) was the highest (42.9 MPa) among all the composite cements. Furthermore, the ZNMPCs were cultured with mouse bone marrow mesenchymal stem cells (mBMSCs). The results yielded that the ZNMPCs exhibited good cytocompatibility with enhanced differentiation, proliferation, and mineralization on mBMSCs, and it also pronouncedly elevated the expressions of genes and proteins involving osteogenesis. These findings suggested that ZNMPCs could drive the differentiation toward osteogenesis and mineralization of mBMSCs, providing a simple way to the MPC with enhanced osteogenesis for further orthopedic applications.

The Fate and Intermediary Metabolism of Soyasapogenol in the Rat.

Research suggests that soyasaponins are poorly absorbed in the GI tract and that soyasaponin aglycones or soyasapogenols are absorbed faster and in greater amounts than the corresponding soyasaponins. Therefore, it is important to understand the bioavailability of these compounds for the potential development of functional foods containing their components. In this paper, to investigate the metabolic characteristics of soyasapogenols A and B, the pharmacokinetic parameters in rats were determined via oral and intravenous administration. The liver metabolites of soyasapogenols were identified using UPLC-/Q-TOF-MS/MS, and their metabolic pathways were also speculated. The results show that, after oral administration, there was a bimodal phenomenon in the absorption process. Tmax was about 2 h, and soyasapogenol was completely metabolized 24 h later. The bioavailability of soyasapogenol was superior, reaching more than 60%. There were sixteen metabolites of soyasapogenol A and fifteen metabolites of soyasapogenol B detected in rat bile. Both phase I and II metabolic transformation types of soyasapogenols, including oxidation, dehydrogenation, hydrolysis, dehydration, deoxidization, phosphorylation, sulfation, glucoaldehyde acidification, and conjugation with cysteine, were identified. In addition, soyasapogenol A could be converted into soyasapogenols B and E in the metabolic process. These results suggest that it is feasible to use soyasapogenols as functional ingredients in nutraceuticals or food formulations.

Effects of bovine cancellous bone powder/poly amino acid composites on cellular behaviors and osteogenic performances.

Xenogeneic bone has good biological activity, but eliminating immunogenicity, while retaining osteogenic abilities, is a challenge. By combining xenogeneic bone with poly amino acid (PAA) that has an amide bond structure, a new type of composite conforming to bionics and low immunogenicity may be obtained. In this study, according to the principles of component bionics, three composites of delipidized cancellous bone powder (DCBP) and PAA were designed and obtained by anin situpolycondensation method, an extrusion molding (EM) method, and a solution-blend method. The three composites were all macroscopically uniform, non-cytotoxic, and demonstrated low immunogenicity by effective removal of residual antigens during preparation. Compared with PAA, mouse bone marrow mesenchymal stem cells (BMSCs) on the surfaces of three composites showed different cellular morphologies. The effects of different preparation methods and cellular morphology on cellular differentiation were confirmed by alkaline phosphatase activity, calcium nodule formation and the expression levels of osteogenic differentiation-related genes (bone morphogenetic protein 2, runt-related transcription factor 2, osteopontin and osteocalcin). Among these composites, DCBP/PAA EM showed best cell proliferation and osteogenic differentiationin vitro, and possessed greater bone formation than PAA in a rabbit femoral condyle study. This study may provide a new method for preparing bioactive bone repair materials with low immunogenicity and superior ability to stimulate differentiation of BMSCsin vitroand osteogenesisin vivo. DCBP/PAA EM might be a promising bone repair material for bone defect treatment.

A novel degradable tricalcium silicate/calcium polyphosphate/polyvinyl alcohol organic-inorganic composite cement for bone filling.

In this study, tricalcium silicate (C3S) calcium/polyphosphate/polyvinyl alcohol organic-inorganic self-setting composites were successfully designed. A variety of tests were conducted to characterize their self-setting properties, mechanical properties, degradation properties, and related biological properties. The composite bone cements showed a short setting time (5.5-37.5 min) with a 5:5-6:4 ratio of C3S/CPP to maintain a stable compressive strength (28 MPa). In addition, PVA effectively reduced the brittleness of the inorganic phase. Degradation experiments confirmed the sustainable surface degradation of bone cement. A maximum degradation rate of 49% was reached within 56 days, and the structure remained intact without collapse. Culturing MC3T3 cells with bone cement extracts revealed that the composite bone cements had excellent biological properties in vitro. The original extract showed a proliferation promotion effect on cells, whereas most of the other original extracts of degradable bone cements were toxic to the cells. Meanwhile, extracellular matrix mineralization and alkaline phosphatase expression showed remarkable effects on cell differentiation. In addition, a good level of adhesion of cells to the surfaces of materials was observed. Taken together, these results indicate that C3S/CPP/PVA composite bone cements have great potential in bone defect filling for fast curing.

Developing a novel magnesium calcium phosphate/sodium alginate composite cement with high strength and proper self-setting time for bone repair.

In this work, novel magnesium calcium phosphate/sodium alginate composite cements were successfully fabricated with a proper setting time (5-24 min) and high compressive strength (91.1 MPa). The physicochemical and biological properties of the cement in vitro were fully characterized. The composite cements could gradually degrade in PBS as the soaking time increase, and the weight loss reached 20.74% by the end of 56th day. The cements could induce the deposition of Ca-P layer in SBF. Cell experiments proved that the extracts of the composite cements can effectively promote the proliferation and differentiation of the mouse bone marrow mesenchymal stem cells (MSCs). These preliminary results indicate that the magnesium calcium phosphate/sodium alginate composite cements could be promising as potential bone repair candidate materials.

Experimental study on repair of large segmental bone defects of goat femur by nano calcium-deficient hydroxyapatite-multi (amino acid) copolymer membrane tubes.

OBJECTIVE: The purpose of this study was to observe feasibility of nano calcium-deficient hydroxyapatite-multi (amino acid) copolymer (n-CDHA-MAC) membrane tubes in repairing goat femurs' large defects. METHODS: Twelve goats were divided into two groups, whose femurs were created 30 mm segmental bone defects and then implants were performed. In experimental group, the bone defect of right femur was reconstructed by n-CDHA-MAC membrane tube, while left side was reconstructed by allogenic bone tube in control group. Every three goats were sacrificed at 4, 8, 16, 24 weeks after operation respectively. General observation, X-ray analysis, histology, Scanning electron microscope (SEM) examination and protein level comparison of BMP-2 were conducted to evaluate the effects of repairing segmental bone defects. RESULTS: All goats recovered well from anesthesia and surgical interventions. The radiographic evaluations showed that periosteal reaction outside of the membrane tubes and allogenic bone tubes were observed 4 weeks after surgery. At 16 weeks, callus was continuously increased in experimental group, which was more obvious than control group. At 24 weeks, callus outside of the membrane tubes connected together. Histologic evaluation showed fibro-cartilage callus was evolved into bony callus in experimental group, which was more obvious than control group at 8 and 16 weeks. The protein expression level of BMP-2 increased at 4, 8 weeks and peaked at 16 weeks in experimental groups. There were statistical differences at 8 and 16 weeks (P < 0.05). At each time point in 8, 16, 24 weeks after surgery, the bending stiffness, torsional stiffness and compressive strength of the two groups were similar, and there was no significant difference (P > 0.05). CONCLUSIONS: This novel surface degradation n-CDHA-MAC membrane tube has good ability to maintain enough membrane space, which can provide long-term and stable biomechanical support for large bone defects and integrate well with the surrounding bone.

Biological evaluation of the modified nano-amorphous phosphate calcium doped with citrate/poly-amino acid composite as a potential candidate for bone repair and reconstruction.

Large numbers of research works related to fabricating organic-inorganic composite materials have been carried out to mimic the natural structure of bone. In this study, a new modified n-ACP doped with citrate (n-ACP-cit)/poly (amino acids) (PAA) composite (n-ACP-cit/PAA) was synthesized by employing high bioactive n-ACP-cit and the biodegradable and biocompatible PAA copolymer. Its basic structure was characterized by X-ray diffraction spectroscopy, Fourier transformed infrared spectroscopy, and X-ray photoelectron spectroscopy. Moreover, the degradability, bioactivity, biocompatibility, and osteoconductivity of n-ACP-cit/PAA composite were evaluated in vitro and in vivo, using simulated body fluid (SBF) solution soaking test, mouse bone marrow mesenchymal stem cells proliferation and differentiation, morphological observation test, expression of genes associated with osteogenesis, and bone defect model repair test, respectively. The modified n-ACP-cit/PAA composite exhibited a much higher weight loss rate (36.01 wt.%) than that of PAA (23.99 wt.%) after immersing in SBF solution for 16 weeks and the pH values of local environment restored to neutral condition. Moreover, cells co-culturing with composites exhibited higher alkaline phosphatase activity, more calcium nodule-formation, and higher expression levels of osteogenic differentiation-related genes (Bmp-2, Colla I, OCN, OPN, and Runx-2) than that of PAA. Furthermore, the bone defect model repair test revealed that the composite could be intimately incorporated with the surrounding bone without causing any deleterious reaction and capable of guiding new bone formation. Together, these results indicated that the new modified bone repair n-ACP-cit/PAA composite material with specific characteristics may be designed for meeting diverse requirements from biomedical applications.

Preparation of a novel bovine cancellous bone/poly-amino acid composite with low immunogenicity, proper strength, and cytocompatibility in vitro.

In this work, the delipidized and deproteinized bovine cancellous bone powder/poly-amino acid (DDBP/PAA) composite was fabricated by extrusion-injection molding method for the first time. After about 70% clearance rate by the delipidization and deproteinization procedures, the residual antigens of galactosyl α-(1, 3)-galactosyl ß-1,4-N-aeetylglueosaminyl (α-Gal) and major histocompatibility complex (MHC) II were basically eliminated by the extrusion-injection molding process, which may cause high titer of antibody and lead to hyperacute rejection or chronic immune toxicity. Meanwhile, the natural BMP II and apatite in bovine bone were kept in DDBP/PAA composite. After 26 weeks of immersion in simulated body fluid, the DDBP/PAA composite remained the intact appearance, 96.4% of weight, and 69.2% of compressive strength, and these showed sufficient degradation stability. The composite also exhibited excellent attachment and proliferation abilities of mouse bone marrow mesenchymal stem cells (mMSCs). The results herein suggested that the DDBP/PAA composite was expected to be a load-bearing transplant with some natural ingredients for hard tissue repair.

Taoren-dahuang herb pair reduces eicosanoid metabolite shifts by regulating ADORA2A degradation activity in ischaemia/reperfusion injury rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Peach kernel (taoren: TR) is the dried mature seed of peach, Prunus persica (L.) Batsch, which belongs to the Rosaceae family. Rhubarb (dahuang: DH) is the dried root and rhizome of rhubarb (Rheum palmatum L., Rheum officinale Baill., or Rheum tanguticum Maxim. ex Balf.). TR-DH (TD) is a traditional Chinese medicine herb pair that promotes blood circulation and removes blood stasis. In recent years, TD has shown definite benefits in the cardio-cerebrovascular system, but its specific mechanism is not very clear. AIM OF STUDY: The purpose of this study was to explore the mechanism by which TD affects cerebral ischaemia/reperfusion (I/R) injury and to optimize the mixture ratio. METHODS: The affected metabolic pathways in rat brain tissues after I/R were analysed by network pharmacology and verified with animal pharmacological experiments. RESULTS: TD had a certain therapeutic effect on cerebral I/R injury. TD with a TR:DH ratio of 1:1 had the best therapeutic effect. Metabolic pathway analysis showed that the protective mechanism of TD against I/R injury involves mainly regulation of brain tissue ADORA2A protein levels and action on the arachidonic acid (AA) pathway. CONCLUSION: TD can ameliorate cerebral I/R injury by regulating ADORA2A degradation in the AA metabolic pathway to attenuate AA metabolic dysfunction and the inflammatory response.

Absorbable nanocomposites composed of mesoporous bioglass nanoparticles and polyelectrolyte complexes for surgical hemorrhage control.

Absorbable polyelectrolyte complexes-based hemostats are promising for controlling hemorrhage in iatrogenic injuries during surgery, whereas their hemostatic efficacy and other performances require further improvement for clinical application. Herein, spherical mesoporous bioglass nanoparticles (mBGN) were fabricated, and mBGN-polyelectrolyte complexes (composed of carboxymethyl starch and chitosan oligosaccharide) nanocomposites (BGN/PEC) with different mBGN contents were prepared via in situ coprecipitation followed by lyophilization. The effect of various mBGN content (10 and 20 wt%) on morphology, zeta potential, water absorption, degradation behavior and ion release were systematically evaluated. The in vitro degradability was dramatically promoted and a more neutral environment was achieved with the incorporation of mBGN, which is preferable for surgical applications. The in vitro coagulation test with whole blood demonstrated that the incorporation of mBGN facilitated blood clotting process. The plasma coagulation evaluation indicated that BGN/PEC had increased capability to accelerate coagulation cascade via the intrinsic pathway than that of the PEC, while have inapparent influence on the extrinsic and common pathway. The in vivo hemostatic evaluation in a rabbit hepatic hemorrhage model revealed that BGN/PEC with 10 wt% mBGN (10BGN/PEC) treatment group had the lowest blood loss, although its hemostatic time is close to that of 20BGN/PEC treatment group. The cytocompatibility evaluation with MC3T3-L1 fibroblasts indicated that 10BGN/PEC induced a ~25% increase of cell viability compared to the PEC at days 4 and 7, indicating improved biocompatibility. These findings support the promising application of absorbable BGN/PEC with optimized mBGN content as internal hemostats and present a platform for further development of PEC-based hemostats.

Virtual Screening of the Multi-pathway and Multi-gene Regulatory Molecular Mechanism of Dachengqi Decoction in the Treatment of Stroke Based on Network Pharmacology.

BACKGROUND: Stroke is ranked second among diseases that cause mortality worldwide. Owing to its complicated pathogenesis, no satisfactory treatment strategies for stroke are available. Dachengqi decoction (DCQD), a traditional Chinese herbal medicine, has been widely used in China for a long time, as it has a good effect on stroke. However, the molecular mechanism underlying this effect of DCQD is unclear. OBJECTIVE: In the present study, we aimed to reveal and explore the multi-pathway and multi-gene regulatory molecular mechanism of Dachengqi decoction in the treatment of stroke. METHODS: In this study, a network pharmacology method, in combination with oral bioavailability prediction and drug-likeness evaluation, was employed to predict the active ingredients of DCQD. The target genes of the active components and the traced pathways related to these target genes were predicted. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed using clusterProfiler software package on the R platform and ClueGo+CluePedia plug-ins. Finally, the key DCQD targets were verified using the Gene Expression Omnibus (GEO) dataset. RESULTS AND DISCUSSION: According to the ADME model, 52 active components were screened from 296 active components of DCQD. After prediction and screening, 215 stroke-related targets were obtained and analyzed via GO and KEGG analyses. GO analysis showed that DCQD targets were mainly involved in the regulation of oxidative stress, lipid metabolism, inflammation, and other biological processes. KEGG pathway analysis further revealed pathways involved in stroke, such as arachidonic acid metabolic, HIF-1 signaling pathway, estrogen signaling pathway, MAPK signaling pathway, PI3K-Akt signaling pathway, platelet activation pathway, VEGF signaling pathway, and cAMP signaling pathway. Network analysis revealed that DCQD might be involved in the regulation of lipid metabolism, blood pressure, inflammation, angiogenesis, neuroprotection, platelet aggregation, apoptosis, and oxidation in stroke treatment. GEO dataset analysis showed that DCQD's therapeutic effects might be exerted via the bidirectional regulation principle. CONCLUSION: Based on the methods of network pharmacology and GEO analysis, it was found that, during stroke treatment, DCQD regulates and controls multiple genes and multiple pathways in a synergistic manner, providing a new strategy for stroke treatment.

Design of novel organic-inorganic composite bone cements with high compressive strength, in vitro bioactivity and cytocompatibility.

In this work, novel bioactive organic-inorganic composite bone cements consisting of tricalcium silicate (C3 S), sodium alginate (SA), and calcium sulfate hemihydrate (CS) were successfully fabricated for the first time via a special method designing material composition and internal structure simultaneously, which was intended to enhance mechanical performance by combining progressive hydration process of C3 S with distinctive gelation capacity of SA and further improve degradability and self-setting properties with the addition of CS. Depending on the synergistic combination of hydration and gelation, the C3 S/SA/CS composite cements (45/45/10 wt %) obtained extremely higher compressive strength up to 92.41 MPa as compared with each single component. The reinforcing mechanisms involving interfacial interaction and interior microstructure were proposed to explain this enhancement phenomenon. Additionally, the final setting time could be reduced from 68 min to 21 min with the increasing CS content. The composite cements possessed good apatite mineralization ability in simulated body fluid solution and moderate degradation rate in phosphate buffer solution. What's more, the composite cements exhibited excellent cytocompatibility and increased proliferation of rat bone-marrow stem cells. This study could provide guidelines for the preparation of bioactive composite cements with enhanced mechanical performance, which may be suitable for load-bearing bone repair. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2365-2377, 2019.

Evaluation of the internal fixation effect of nano-calcium-deficient hydroxyapatite/poly-amino acid composite screws for intraarticular fractures in rabbits.

OBJECTIVE: To evaluate the internal fixation effect of nano-calcium-deficient hydroxyapatite/poly-amino acid (n-CDHA/PAA) composite screws in the intraarticular fracture model. MATERIALS AND METHODS: A total of 35 New Zealand White rabbits were used in a bilateral femoral intercondylar fracture model and randomly divided into two groups. n-CDHA/PAA screws were used in the experimental group, and medical metal screws were used in the control group. The fracture condition, range of motion, and the screw push-out strength were assessed, and an arthroscopic examination of knee joint was performed at 4, 8, and 12 weeks after surgery. The biodegradation of the n-CDHA/PAA screws in vivo was tested through weighing, and changes in screw structure were assessed by X-ray diffraction at 12 weeks after surgery. RESULTS: The general situation of all animals was good and showed no incision infection and dehiscence after surgery. X-ray scanning showed that significant callus growth was present in both groups at 4 weeks after surgery, and there was no significant difference (P>0.05) in the Lane-Sandhu score between the experimental and control groups at all time points after surgery. There were no statistically significant differences (P>0.05) in the range of motion and Oswestry Arthroscopy Score of arthroscopic examination of the knee joints between the two groups. The screw push-out strength of the control group was stronger than that of the experimental group at 4 weeks after surgery (P<0.05), but after that, there was no significant difference between the groups (P>0.05). The degradation tests showed that the n-CDHA/PAA screws degraded gradually after implantation, and the weight loss rate was approximately 16% at 12 weeks after surgery. The X-ray diffraction results showed that the crystal structure of the outer surface of the n-CDHA/PAA screw has changed at 12 weeks after surgery. CONCLUSION: The n-CDHA/PAA screw is an effective and safe implant as a potential internal fixation device for an intercondylar fracture of the femur, and its internal fixation effect was similar to that of medical metal screw.

Developing novel Ca-zeolite/poly(amino acid) composites with hemostatic activity for bone substitute applications.

The novel Ca-zeolite/poly(amino acid) (CaY/PAA) composites for bone substitute applications with hemostatic activity were prepared using the in situ melting polymerization method. In this study, Ca-zeolite (CaY) loaded with Ca2+ was obtained from Y-type zeolite (NaY) by ion-exchange method. The properties of the CaY/PAA composites and PAA, including composition, structure, compressive strength, in vitro degradability in phosphate-buffered solution (PBS), bioactivity, cytocompatibility and in vitro coagulation tests were characterized and investigated. The results showed that compressive strength of the CaY/PAA composites ranged from 145 to 186 MPa, demonstrating sufficient mechanical strength for load-bearing bone substitute. After soaking in PBS for 16 weeks, the weight loss of 25CaY/PAA and 50CaY/PAA were 4.1 and 1.6 wt%, respectively, and the pH values for CaY/PAA composites increased to about 8.0 in 2 weeks and then gradually stabilized around 7.4, indicating good stability in PBS. Scanning electron microscope and energy dispersive spectrometer results showed that the composites were bioactive and new apatite layers attached on their surfaces. Mesenchymal stem cells (MSCs) exhibited high-proliferation in the extract solution of the CaY/PAA composites and were well spread on the surfaces of the composites. Cells on the CaY/PAA composite groups showed higher alkaline phosphatase (ALP) activity indicating the potential to promote cell differentiation. The in vitro coagulation tests showed that CaY/PAA composites have shorter clotting time and better performance of promoting blood coagulation than other samples, presenting improved hemostatic activity. In summary, the results demonstrated that the CaY/PAA composites had good mechanical strength, stability, bioactivity, cytocompatibility and hemostatic activity for bone substitute applications.