Excellent Apatite-mineralization Ability Research Articles

Design and Evaluation of Europium Containing Mesoporous Bioactive Glass Nanospheres: Doxorubicin Release Kinetics and Inhibitory Effect on Osteosarcoma MG 63 Cells.

Functional ions and drug factors play a vital role in stimulating bone tissue regeneration as we understand it. In this work, europium-containing mesoporous bioactive glass nanospheres (Eu/MBGs), composed of 60% SiO2—(36–x)%CaO—x%Eu2O3—4%P2O5 (x = 0, 0.5, 1, 2 mol%), were prepared by a facile sol-gel process. The results indicate that Eu ions play an important role to influence the microstructure of MBGs, in which a suitable concentration of Eu (1 mol%) increases their surface area (502 m2/g) as well as their pore volume (0.34 cm3/g). Proper doping of Eu ions in MBGs can observably induce apatite mineralization and improve the doxorubicin (DOX) release behavior. Furthermore, DOX-loaded Eu/MBGs could maintain a long-term inhibitory effect on the viability of osteosarcoma MG 63 cells. This work has demonstrated that it is possible to develop functional Eu/MBGs by combining excellent apatite-mineralization ability, controllable drug (DOX) release and antitumor functions for the therapy of bone tissue regeneration.

3D printed porous β-Ca2SiO4 scaffolds derived from preceramic resin and their physicochemical and biological properties

ABSTRACTSilicate bioceramic scaffolds are of great interest in bone tissue engineering, but the fabrication of silicate bioceramic scaffolds with complex geometries is still challenging. In this study, three-dimensional (3D) porous β-Ca2SiO4 scaffolds have been successfully fabricated from preceramic resin loaded with CaCO3 active filler by 3D printing. The fabricated β-Ca2SiO4 scaffolds had uniform interconnected macropores (ca. 400 μm), high porosity (>78%), enhanced mechanical strength (ca. 5.2 MPa), and excellent apatite mineralization ability. Importantly, the results showed that the increase of sintering temperature significantly enhanced the compressive strength and the scaffolds sintered at higher sintering temperature stimulated the adhesion, proliferation, alkaline phosphatase activity, and osteogenic-related gene expression of rat bone mesenchymal stem cells. Therefore, the 3D printed β-Ca2SiO4 scaffolds derived from preceramic resin and CaCO3 active fillers would be promising candidates for bone tissue engineering.

Zein regulating apatite mineralization, degradability, in vitro cells responses and in vivo osteogenesis of 3D-printed scaffold of n-MS/ZN/PCL ternary composite.

Bioactive and degradable scaffolds of nano magnesium silicate (n-MS)/zein (ZN)/poly(caprolactone) (PCL) ternary composites were prepared by 3D-printing method. The results showed that the 3D-printed scaffolds possessed controllable pore structure, and pore morphology, pore size, porosity and pore interconnectivity of the scaffolds can be efficiently adjusted. In addition, the apatite-mineralization ability of the scaffolds in simulated body fluids was obviously improved with the increase of ZN content, in which the scaffold with 20 w% ZN (C20) possessed excellent apatite-mineralization ability. Moreover, the degradability of the scaffolds was significantly enhanced with the increase of ZN content in the scaffolds. The degradation of ZN produced acidic products that could neutralize the alkaline products from the degradation of n-MS, which avoid the increase of pH value in degradable solution. Furthermore, the MC3T3-E1 cells responses (e.g. proliferation and differentiation, etc.) to the scaffolds were significantly promoted with the increase of ZN content. The in vivo osteogenesis of the scaffolds implanted the femur defects of rabbits was investigated by micro-CT and histological analysis. The results demonstrated that the new bone formation was significantly enhanced with the increase of ZN content, in which the C20 scaffold induced the highest new bone tissues, indicating excellent osteogenesis. The results suggested that the ZN in the ternary composite scaffolds played key roles in assisting bone regeneration in vivo.

Effects of mesoporous bioglass on physicochemical and biological properties of calcium sulfate bone cements

Bone cement is a promising substitute for bone repair due to its simple operation, excellent plasticity and self-setting after filling bone defect by injection. In this study, calcium sulfate hydrate (CSH)/mesoporous bioglass (MBG) bone cements were prepared through changing the MBG incorporation amount. The ratio of liquid to powder (RLP), mechanical strength, in vitro bioactivity, degradation and cell responses on CSH/MBG cements were investigated. The results showed that appropriate setting times for CSH/MBG cements could be achieved by adjusting the RLP, and the compressive strength was in the range of 5–16MPa. Also, CSH/MBG cements had excellent apatite mineralization ability, and the MBG incorporation could regulate the degradation rate. Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured on CSH/MBG cements, and the results indicated that CSH/MBG cements stimulated cell proliferation and differentiation. Furthermore, CSH/MBG cements had a sustained drug release property for use in local drug delivery therapy. Hence, CSH/MBG cements have more potential to be bone substitute for bone repair.

Three dimensional printing of calcium sulfate and mesoporous bioactive glass scaffolds for improving bone regeneration in vitro and in vivo

In the clinic, bone defects resulting from infections, trauma, surgical resection and genetic malformations remain a significant challenge. In the field of bone tissue engineering, three-dimensional (3D) scaffolds are promising for the treatment of bone defects. In this study, calcium sulfate hydrate (CSH)/mesoporous bioactive glass (MBG) scaffolds were successfully fabricated using a 3D printing technique, which had a regular and uniform square macroporous structure, high porosity and excellent apatite mineralization ability. Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured on scaffolds to evaluate hBMSC attachment, proliferation and osteogenesis-related gene expression. Critical-sized rat calvarial defects were applied to investigate the effect of CSH/MBG scaffolds on bone regeneration in vivo. The in vitro results showed that CSH/MBG scaffolds stimulated the adhesion, proliferation, alkaline phosphatase (ALP) activity and osteogenesis-related gene expression of hBMSCs. In vivo results showed that CSH/MBG scaffolds could significantly enhance new bone formation in calvarial defects compared to CSH scaffolds. Thus 3D printed CSH/MBG scaffolds would be promising candidates for promoting bone regeneration.

Three-dimensional printing of tricalcium silicate/mesoporous bioactive glass cement scaffolds for bone regeneration.

Bone defects, particularly large bone defects resulting from infections, trauma, surgical resection or genetic malformations, remain a significant challenge for clinicians. In this study, the tricalcium silicate/mesoporous bioactive glass (C3S/MBG) cement scaffolds were successfully fabricated for the first time by 3D printing with a curing process, which combined the hydraulicity of C3S with the excellent biological property of MBG together. The C3S/MBG scaffolds exhibited 3D interconnected macropores (∼400 μm), high porosity (∼70%), enhanced mechanical strength (>12 MPa) and excellent apatite mineralization ability. Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured on the scaffolds to evaluate their cell responses, and the results showed that C3S/MBG scaffolds could stimulate the attachment, proliferation and differentiation of hBMSCs with increasing MBG component. The critical-sized rat calvarial defect animal model was employed; further in vivo results indicated that both C3S and C3S/MBG30 scaffolds could induce new bone formation, but the C3S/MBG30 scaffolds significantly improved the osteogenic capacity compared to the pure C3S scaffolds. Therefore, the C3S/MBG cement scaffolds fabricated by 3D printing with a curing process would be a promising candidate for bone regeneration.

Silicate bioceramic/PMMA composite bone cement with distinctive physicochemical and bioactive properties

New bioactive silicate/PMMA composite bone cements possess improved setting properties, high mechanical strength, excellent apatite-mineralization ability and biological activity for injectable bone regeneration materials application.

Bioactive bredigite coating with improved bonding strength, rapid apatite mineralization and excellent cytocompatibility

Previous studies have shown that bredigite (Ca7MgSi4O16) bioceramics possessed excellent biocompatibility, apatite-mineralization ability and mechanical properties. In this paper, the bredigite coating on Ti-6Al-4 V substrate was prepared by plasma spraying technique. The main compositions of the coating were bredigite crystal phase with small parts of amorphous phases. The bonding strength of the coating to Ti-6Al-4 V substrate reached 49.8 MPa, which was significantly higher than that of hydroxyapatite coating and other silicate-based bioceramic coatings prepared by same method. After immersed in simulated body fluid for 2 days, a distinct apatite layer was deposited on the surface of bredigite coating, indicating that the prepared bredigite coating has excellent apatite-mineralization ability. The prepared bredigite coating supported the attachment and proliferation of rabbit bone marrow stem cells. The proliferation level of bone marrow stem cells was significantly higher than that on the hydroxyapatite coating. Our further study showed that the released SiO4 (4-) and Mg(2+) ions from bredigite coating as well as the formed nano-apatite layer on the coating surface might mainly contribute to the improvement of cell proliferation. The results indicated that the bredigite coating may be applied on orthopedic implants due to its excellent bonding strength, apatite mineralization and cytocompatibility.

In vitro assessment of three-dimensionally plotted nagelschmidtite bioceramic scaffolds with varied macropore morphologies

It is known that porous scaffolds play an important role in bone/periodontal tissue engineering. A new nagelschmidtite (NAGEL, Ca7Si2P2O16) ceramic has recently been prepared which shows excellent apatite mineralization ability and osteo-/cementostimulation properties in vitro. However, up to now porous NAGEL scaffolds have not been developed yet. There has been no systematic study of the effect of macropore morphology of bioceramic scaffolds on their physico-chemical and biological properties. The aim of this study was to prepare NAGEL scaffolds for bone tissue engineering applications. We applied a modified three-dimensional (3-D) plotting method to prepare highly controllable NAGEL scaffolds and investigated the effect of macropore morphology on the physico-chemical and biological properties. The results showed that the macropore size and morphology of 3-D plotted NAGEL scaffolds could be effectively controlled. Compared with β-tricalcium phosphate (β-TCP) scaffolds NAGEL scaffolds possess a significantly enhanced compressive strength, a higher modulus and better degradability. Nagel scaffolds with a square pore morphology presented a higher compressive strength, a higher modulus and greater weight loss rate than those with triangular and parallelogram pore morphologies. In addition, all of the NAGEL scaffolds with the three macropore morphologies supported the attachment and proliferation of MC3T3 cells. The proliferation of MC3T3 cells on NAGEL scaffolds with triangular and parallelogram structures was higher than that on β-TCP scaffolds with the same pore structure. Cells on all three groups of NAGEL scaffolds revealed higher alkaline phosphatase (ALP) activity compared with cells on β-TCP scaffolds, and among the three NAGEL scaffolds groups those with a parallelogram pore structure showed the highest ALP activity. Furthermore, the angiogenic cell experiments showed that the ionic products from NAGEL scaffolds promoted tube formation, expression of pro-angiogenic factors and their receptors on human umbilical vein endothelial (HUVECs) compared with β-TCP scaffolds, indicating that NAGEL scaffolds possessed improved angiogenesis capacity. Our results suggest that 3-D plotted NAGEL scaffolds are a promising bioactive material for bone tissue engineering by virtue of their highly controllable macropore structure, excellent mechanical strength, degradability and in vitro biological response to osteogenic/angiogenic cells.

Functional mesoporous bioactive glass nanospheres: synthesis, high loading efficiency, controllable delivery of doxorubicin and inhibitory effect on bone cancer cells.

Controllable drug delivery is one of the important ways for the therapy of bone cancer. Conventional mesoporous silica nano-particles may lack dual properties for combining controllable delivery of anti-cancer drugs and bone-forming bioactivity for bone cancer therapy. The aim of this study is to synthesize mesoporous bioactive glass (MBG) nanospheres with combined dual functions of bioactivity and controlled drug delivery, and to further investigate their delivery property of anti-cancer drugs as well as the functional effect on bone-cancer cells. MBG nanospheres with spherical morphology and internal mesoporous microstructures were successfully prepared by a facile hydrothermal method. The prepared MBG nanospheres possess high specific surface area and mesopore volume (443 m2 g-1, 0.57 cm3 g-1) as well as uniform mesopore size distribution (2.9 nm). The MBG nanospheres demonstrate excellent bioactivity by inducing apatite mineralization in simulated body fluids. An anti-cancer drug, doxorubicin hydrochloride (DOX), was successfully loaded in the MBG nanospheres with a distinctively high loading efficiency of around 90%. The loading amount of DOX can be effectively controlled by adjusting the initial drug-loading concentrations. MBG nanospheres can maintain a sustained release of DOX, and their release kinetics can be controlled by varying the pH microenvironment and initial drug-loading concentrations. In addition, the prepared MBG nanospheres showed obvious degradation by releasing Ca2+ and SiO4 4- ions in PBS. Furthermore, the delivery of DOX from MBG nanospheres into cell culture environment shows a significant inhibitory effect on the viability of osteosarcoma cells with the increase of interaction time. The prepared MBG nanospheres have high specific surface area and mesopore volume, excellent apatite-mineralization ability, distinct degradability, high DOX-loading efficiency and controllable DOX release as well as anti-cancer functions. These unique characteristics suggest that the obtained MBG nanospheres may be used for the therapy of bone cancer.

Nagelschmidtite bioceramics with osteostimulation properties: material chemistry activating osteogenic genes and WNT signalling pathway of human bone marrow stromal cells.

Bioactive materials with osteostimulation properties are of great importance to promote osteogenic differentiation of human bone marrow stromal cells (hBMSCs) for potential bone regeneration. We have recently synthesized nagelschmidtite (NAGEL, Ca7Si2P2O16) ceramic powders which showed excellent apatite-mineralization ability. The aim of this study was to investigate the interaction of hBMSCs with NAGEL bioceramic bulks and their ionic extracts, and to explore the osteostimulation properties of NAGEL bioceramics and the possible molecular mechanism. The cell attachment, proliferation, bone-related gene expression (ALP, OPN and OCN) and WNT signalling pathways (WNT3a, FZD6, AXIN2 and CTNNB) of hBMSCs cultured on NAGEL bioceramic disks were systematically studied. We further investigated the biological effects of ionic products from NAGEL powders on cell proliferation and osteogenic differentiation of hBMSCs by culturing cells with NAGEL extracts. Furthermore, the effect of NAGEL bioceramics on the osteogenic differentiation in hBMSCs was also investigated with the addition of cardamonin, a WNT inhibitor. The results showed that NAGEL bioceramic disks supported the attachment and proliferation of hBMSCs, and significantly enhanced the bone-related gene expression and WNT signalling pathway of hBMSCs, compared to conventional beta-tricalcium phosphate (β-TCP) bioceramic disks and blank controls. The ionic products from NAGEL powders also significantly promoted the proliferation, bone and WNT-related gene expression of hBMSCs. It was also identified that NAGEL bioceramics could bypass the action of the WNT inhibitor (10 μM) to stimulate the selected osteogenic genes in hBMSCs. Our results suggest that NAGEL bioceramics possess excellent in vitro osteostimulation properties. The possible mechanism for the osteostimulation may be directly related to the released Si, Ca and P-containing ionic products from NAGEL bioceramics which activate bone-related gene expression and WNT signalling pathway of hBMSCs. The present study suggests that NAGEL bioceramics are a potential bone regeneration material with significant osteostimulation capacity.

Strontium-containing mesoporous bioactive glass scaffolds with improved osteogenic/cementogenic differentiation of periodontal ligament cells for periodontal tissue engineering

To achieve the ultimate goal of periodontal tissue engineering, it is of great importance to develop bioactive scaffolds which can stimulate the osteogenic/cementogenic differentiation of periodontal ligament cells (PDLCs) for the favorable regeneration of alveolar bone, root cementum and periodontal ligament. Strontium (Sr) and Sr-containing biomaterials have been found to induce osteoblast activity. However, there has been no systematic report about the interaction between Sr or Sr-containing biomaterials and PDLCs for periodontal tissue engineering. The aims of this study were to prepare Sr-containing mesoporous bioactive glass (Sr-MBG) scaffolds and investigate whether the addition of Sr could stimulate osteogenic/cementogenic differentiation of PDLCs in a tissue-engineering scaffold system. The composition, microstructure and mesopore properties (specific surface area, nanopore volume and nanopore distribution) of Sr-MBG scaffolds were characterized. The proliferation, alkaline phosphatase (ALP) activity and osteogenesis/cementogenesis-related gene expression (ALP, Runx2, Col I, OPN and CEMP1) of PDLCs on different kinds of Sr-MBG scaffolds were systematically investigated. The results show that Sr plays an important role in influencing the mesoporous structure of MBG scaffolds in which high contents of Sr decreased the well-ordered mesopores as well as their surface area/pore volume. Sr2+ ions could be released from Sr-MBG scaffolds in a controlled way. The incorporation of Sr into MBG scaffolds has significantly stimulated ALP activity and osteogenesis/cementogenesis-related gene expression of PDLCs. Furthermore, Sr-MBG scaffolds in a simulated body fluid environment still maintained excellent apatite-mineralization ability. The study suggests that the incorporation of Sr into MBG scaffolds is a viable way to stimulate the biological response of PDLCs. Sr-MBG scaffolds are a promising bioactive material for periodontal tissue-engineering applications.

The stimulation of proliferation and differentiation of periodontal ligament cells by the ionic products from Ca7Si2P2O16 bioceramics

The ultimate goal of periodontal tissue engineering is to produce predictable regeneration of alveolar bone, root cementum, and periodontal ligament, which are lost as a result of periodontal diseases. To achieve this goal, it is of great importance to develop novel bioactive materials which could stimulate the proliferation, differentiation and osteogenic/cementogenic gene expression of periodontal ligament cells (PDLCs) for periodontal regeneration. In this study, we synthesized novel Ca7Si2P2O16 ceramic powders for the first time by the sol–gel method and investigated the biological performance of PDLCs after exposure to different concentrations of Ca7Si2P2O16 extracts. The original extracts were prepared at 200mgml–1 and further diluted with serum-free cell culture medium to obtain a series of diluted extracts (100, 50, 25, 12.5 and 6.25mgml–1). Proliferation, alkaline phosphatase (ALP) activity, Ca deposition, and osteogenesis/cementogenesis-related gene expression (ALP, Col I, Runx2 and CEMP1) were assayed for PDLCs on days 7 and 14. The results showed that the ionic products from Ca7Si2P2O16 powders significantly stimulated the proliferation, ALP activity, Ca deposition and osteogenesis/cementogenesis-related gene expression of PDLCs. In addition, it was found that Ca7Si2P2O16 powders had excellent apatite-mineralization ability in simulated body fluids. This study demonstrated that Ca7Si2P2O16 powders with such a specific composition possess the ability to stimulate the PDLC proliferation and osteoblast/cemenoblast-like cell differentiation, indicating that they are a promising bioactive material for periodontal tissue regeneration application.

The effect of poly(lactic-co-glycolic acid) (PLGA) coating on the mechanical, biodegradable, bioactive properties and drug release of porous calcium silicate scaffolds

Ideal scaffolds for bone tissue engineering require 3D interconnected porous structures, enough mechanical strength for hand of treatment as well as proper bioactivity and biodegradability. Calcium silicate (CaSiO3, CS) scaffolds have been studied for bone tissue engineering application due to their excellent bioactivity. However, the main disadvantages of CS scaffolds are their low mechanical strength and high alkaline ionic products. In this study, sintered CS scaffolds were prepared and coated with poly(lactic-co-glycolic acid) (PLGA), and the effect of PLGA coating on the mechanical, biodegradable, bioactive properties and drug release of porous CS scaffolds were investigated. The results showed that the PLGA-coated CS scaffolds maintained large pore size and high porosity. The compressive strength of PLGA/CS scaffolds was significantly improved compared to pure CS scaffolds, and increased with the increase of intrinsic viscosity and concentration of PLGA. In addition, the PLGA coating neutralized alkaline level resulted from the ionic products of CS scaffolds and reduced the pH value of biological solution during the degradation of scaffolds. It was found that PLGA/CS scaffolds still maintained excellent apatite-mineralization ability in SBF. Furthermore, the PLGA coating effectively inhibited the burst release and maintained a sustained release of drugs from the CS scaffolds. Our results indicate that the PLGA/CS scaffolds have great potential for bone tissue engineering application by the virtue of improved mechanical strength, and excellent bioactivity, degradation as well as drug-delivery property.

3D-printing of highly uniform CaSiO3 ceramic scaffolds: preparation, characterization and in vivo osteogenesis

Calcium silicate (CaSiO3, CS) ceramics have received significant attention for application in bone regeneration due to their excellent in vitro apatite-mineralization ability; however, how to prepare porous CS scaffolds with a controllable pore structure for bone tissue engineering still remains a challenge. Conventional methods could not efficiently control the pore structure and mechanical strength of CS scaffolds, resulting in unstable in vivo osteogenesis. The aim of this study is to set out to solve these problems by applying a modified 3D-printing method to prepare highly uniform CS scaffolds with controllable pore structure and improved mechanical strength. The in vivo osteogenesis of the prepared 3D-printed CS scaffolds was further investigated by implanting them in the femur defects of rats. The results show that the CS scaffolds prepared by the modified 3D-printing method have uniform scaffold morphology. The pore size and pore structure of CS scaffolds can be efficiently adjusted. The compressive strength of 3D-printed CS scaffolds is around 120 times that of conventional polyurethane templated CS scaffolds. 3D-Printed CS scaffolds possess excellent apatite-mineralization ability in simulated body fluids. Micro-CT analysis has shown that 3D-printed CS scaffolds play an important role in assisting the regeneration of bone defects in vivo. The healing level of bone defects implanted by 3D-printed CS scaffolds is obviously higher than that of 3D-printed β-tricalcium phosphate (β-TCP) scaffolds at both 4 and 8 weeks. Hematoxylin and eosin (H&E) staining shows that 3D-printed CS scaffolds induce higher quality of the newly formed bone than 3D-printed β-TCP scaffolds. Immunohistochemical analyses have further shown that stronger expression of human type I collagen (COL1) and alkaline phosphate (ALP) in the bone matrix occurs in the 3D-printed CS scaffolds than in the 3D-printed β-TCP scaffolds. Considering these important advantages, such as controllable structure architecture, significant improvement in mechanical strength, excellent in vivo osteogenesis and since there is no need for second-time sintering, it is indicated that the prepared 3D-printed CS scaffolds are a promising material for application in bone regeneration.

Three-dimensional printing of hierarchical and tough mesoporous bioactive glass scaffolds with a controllable pore architecture, excellent mechanical strength and mineralization ability

New generation biomaterials for bone regeneration should be highly bioactive, resorbable and mechanically strong. Mesoporous bioactive glass (MBG), a novel bioactive material, has been used to study bone regeneration due to its excellent bioactivity, degradation and drug delivery ability, however, the construction of three-dimensional (3-D) MBG scaffolds (as for other bioactive inorganic scaffolds) for bone regeneration remains a significant challenge due to their inherent brittleness and low strength. In this brief communication we report a new facile method to prepare hierarchical and multifunctional MBG scaffolds with a controllable pore architecture, excellent mechanical strength and mineralization ability for application in bone regeneration by a modified 3-D printing technique using polyvinylalcohol (PVA) as a binder. The method provides a new way to solve commonly existing issues for inorganic scaffold materials, for example, uncontrollable pore architectures, low strength, high brittleness and the requirement for a second sintering at high temperature. The 3-D printed MBG scaffolds obtained possess a high mechanical strength about 200 times that of traditional polyurethane foam templated MBG scaffolds. They have a highly controllable pore architecture, excellent apatite mineralization ability and sustained drug delivery properties. Our study indicates that 3-D printed MBG scaffolds may be an excellent candidate for bone regeneration.