Double-network composites based on inorganic fillers reinforced dextran-based hydrogel with high strength.

The biodegradable hydrogels with a 3D network structure have potential applications in bone tissue engineering. Here, inspired by natural bone, the novel organic-inorganic composites (GelMPC-x) with high compressive strength are designed via adding magnesium oxide/calcium dihydrogen phosphate (MPC) powders into the oxidized dextran/gelatin (OD/Gel) hydrogel. GelMPC-x composites can trigger the gelation of OD/Gel hydrogel through an acid-alkaline reaction between magnesium oxide and calcium dihydrogen phosphate, thus forming an organic-inorganic double network. The cross-linked network between oxidized dextran and gelatin, and the multiple weak interactions between OD/Gel hydrogel and MPC enable the composites to have remarkable compressive strength (77-652 kPa) at the strain of 44 %. More importantly, the composites with appropriate MPC content possess superior injectability, high porosity, and excellent cytocompatibility. This work provides guidelines for the preparation of oxidized dextran-based composite hydrogels with enhanced mechanical performance.

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.

Effects of pullulan on the biomechanical and anti-collapse properties of dicalcium phosphate dihydrate bone cement.

In this work, a modified dicalcium phosphate dihydrate (DCPD) bone cement with unique biodegradable ability in a calcium phosphate cement system was prepared by the hydration reaction of monocalcium phosphate monohydrate and calcium oxide and integration with pullulan (Pul), a non-toxic, biocompatible, viscous, and water-soluble polysaccharide that has been successfully used to improve defects in DCPD bone cement, especially its rapid solidification, fragile mechanical properties, and easy collapse. The effect of different contents of Pul on the structure and properties of DCPD were also studied in detail. The modified cement was characterised by X-ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy, ultraviolet-visible absorption, X-ray photoelectron spectroscopy analysis, and rheological property measurements. The results indicated that Pul promoted the hydration formation of DCPD, and interface bonding occurred between Pul and DCPD. With increasing content of Pul, the setting time of the DCPD bone cement increased from 2.6 min to 42.3 min, the compressive strength increased from 0 MPa to 20.4 MPa, and the anti-collapse ability also improved owing to the strong interface bonding, implying that the DCPD bone cement improved by Pul has better potential for application in the field of non-loading bone regenerative medicine compared to unmodified DCPD bone cement.

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.

Developing a biodegradable tricalcium silicate/glucono-delta-lactone/calcium sulfate dihydrate composite cement with high preliminary mechanical property for bone filling.

Bone cements with the feature of easily shaping could ideally match the defect site and prevent the ingrowth of fibrous tissue. In this manuscript, a biodegradable tricalcium silicate (C3S)/glucono-delta-lactone (GDL)/calcium sulfate dihydrate (CSD) organic-inorganic composite cement was fabricated with shorter setting time (less than 15 min) and high preliminary mechanical property (5.27 MPa in the first hour). Many methods were applied to study the physicochemical and biological properties of the cement in vitro. The weight loss in PBS can reach 58% after 12 weeks soaking indicating the better biodegradability. The excellent bioactivity in vitro was emerging after the cement was soaked in the simulated body fluid. The cell experiments showed that suitable concentration of the extract liquid of cement was conducive to the proliferation, differentiation and extracellular matrix calcification of the mouse bone marrow stromal cells. Briefly, the C3S/GDL/CSD composite cement would have the bright capacity for bone filling.

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.

Preparation, Characterization and In Vitro Biological Evaluation of a Novel Pearl Powder/Poly-Amino Acid Composite as a Potential Substitute for Bone Repair and Reconstruction.

Many studies about fabricating organic-inorganic composite materials have been carried out in order to mimic the natural structure of bone. Pearl, which has a special block-and-mortar hierarchical structure, is a superior bone repair material with high osteogenic activity, but it shows few applications in the clinical bone repair and reconstruction because of its brittle and uneasily shaped properties. In this work, pearl powder (P)/poly (amino acid) (PAA) composites were successfully prepared by a method of in situ melting polycondensation to combine the high osteogenic activity of the pearl and the pliability of the PAA. The mechanical properties, in vitro bioactivity and biocompatibility as well as osteogenic activity of the composites were investigated. The results showed that P/PAA composites have both good mechanical properties and bioactivity. The compressive strength, bending strength and tensile strength of the composites reached a maximum of 161 MPa, 50 MPa and 42 MPa, respectively; in addition, apatite particles successfully deposited on the composites surface after immersion in simulated body fluid (SBF) for 7 days indicated that P/PAA composites showed an enhanced mineralization capacity and bioactivity due to incorporation of pearl powder and PAA. The cell culture results revealed that higher cell proliferation and better adhesion morphology of mouse bone marrow mesenchymal stem cells (MSCs) appeared on the composite surface. Moreover, cells growing on the surface of the composites exhibited higher alkaline phosphatase (ALP) activity, more calcium nodule-formation, and higher expression levels of osteogenic differentiation-related genes (COL 1, RunX2, OCN, and OPN) than cells grown on PAA surface. The P/PAA composites exhibited both superior mechanical properties to the pearl powder, higher bioactivity and osteogenic capability compared with those of PAA.

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.

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.

Injectable negatively charged gelatin microsphere-based gels as hemostatic agents for intracavitary and deep wound bleeding in surgery.

Gelatin, as natural macromolecular material, has been used in biomedical fields widely. In this study, various injectable gelatins A, B, and their compound AB microsphere-based gels (A-GMGs, B-GMGs and AB-GMGs) were prepared through water-in-oil emulsion method for hemostasis, and the effects of blood coagulation in vitro and surgical hemostasis (a deep liver wound model) in vivo were evaluated. Furthermore, the influences of gelatin sorts, the size of microsphere, zeta potential (ZP) and viscoelastic properties on hemostasis were also assessed. Results showed that the gelatin microspheres (GMs) exhibited smooth surface, good sphericity and the particle size of a rough normal distribution. GMs carried negative charges and their electronegativity was stronger than that of gelatin A (GA) and gelatin B (GB) raw materials. Rheological analysis showed that a decreasing particle size of the microspheres led to stronger gel strength, and solid-like gels were exhibited under low stress conditions and liquid-like gels were exhibited under high stress conditions. The blood clotting time of B-GMGs was within 60 s, which exhibited a significantly higher blood clotting effect compared with control groups. The hemostasis assay in vivo showed that the gels had better hemostatic effect on a deep liver wound bleeding model compared with control groups, especially B-GMGs. However, in vivo and vitro hemostatic experiments, particle size of GMs had no obvious influence on the hemostatic effect of the gels. In addition, the CCK-8 assay of bone marrow mesenchymal stem cells of murine (mMSCs) indicated non-cytotoxicity of GMs for cells. These results demonstrated that the gelatin microsphere-based gels (GMGs) had potential to be an effective hemostatic material for intracavitary and deep wound bleeding in surgery.

Developing a novel magnesium glycerophosphate/silicate-based organic-inorganic composite cement for bone repair.

Considering that the phospholipids and glycerophosphoric acid are the basic materials throughout the metabolism of the whole life period and the bone is composed of organic polymer collagen and inorganic mineral apatite, a novel self-setting composite of magnesium glycerophosphate (MG) and di-calcium silicate(C2S)/tri-calcium silicate(C3S) was developed as bio-cement for bone repair, reconstruction and regeneration. The composite was prepared by mixing the MG, C2S and C3S with the certain ratios, and using the deionized water and phosphoric acid solution as mixed liquid. The combination and formation of the composites was characterized by FTIR, XPS and XRD. The physicochemical properties were studied by setting time, compressive strength, pH value, weight loss in the PBS and surface change by SEM-EDX. The biocompatibility was evaluated by cell culture in the leaching solution of the composites. The preliminary results showed that when di- and tri-calcium silicate contact with water, there are lots of Ca(OH)2 generated making the pH value of solution is higher than 9 which is helpful for the formation of hydroxyapatite(HA) that is the main bone material. The new organic-inorganic self-setting bio-cements showed initial setting time is ranged from 20 min to 85 min and the compressive strength reached 30 MPa on the 7th days, suitable as the bone fillers. The weight loss was 20% in the first week, and 25% in the 4th week. Meanwhile, the new HA precipitated on the composite surface during the incubation in the SBF showed bioactivity. The cell cultured in the leaching liquid of the composite showed high proliferation inferring the new bio-cement has good biocompatibility to the cells.

The normal modes of lattice vibrations of ice XI.

The vibrational spectrum of ice XI at thermal wavelengths using the CASTEP code, a first-principles simulation method, is investigated. A dual-track approach is constructed to verify the validity for the computational phonon spectrum: collate the simulated spectrum with inelastic neutron scattering experiments and assign the photon scattering peaks according to the calculated normal vibration frequencies. The 33 optical normal vibrations at the Brillouin center are illustrated definitely from the ab initio outcomes. The depolarizing field effect of the hydrogen bond vibrations at frequencies of 229 cm(-1) and 310 cm(-1) is found to agree well with the LST relationship. It is a convincing evidence to manifest the LO-TO splitting of hydrogen bonds in ice crystal. We attribute the two hydrogen bond peaks to the depolarization effect and apply this viewpoint to ordinary ice phase, ice Ih, which is difficult to analyse their vibration modes due to proton disorder.