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ABSTRACT: Currently, nanobioceramics have received much attention due to their potentially high biological performance. In the present study, the interactions between proteins and two types of hydroxyapatite (HA) ceramic particles with distinct microstructures were studied in vitro. Protein adsorption on the microwave sintered and conventionally sintered HA ceramic particles (named as HAMS and HACS, respectively) were carried out in simulated body fluid (SBF) containing model proteins or rat serum and then subjected to protein quantitative evaluation, SDS-PAGE, and Western blotting analysis. The ceramic particles were characterized by nitrogen sorption, Hg penetration, zeta potential, and solubility analysis. It was found that HAMS with nanosized crystallites had greater specific surface area and pore volume and wider pore size distribution ranging from 0.02 to 2 μm than HACS. Although bovine serum albumin and lysozyme have different electrical properties in SBF, both the model proteins showed higher adsorption amounts per gram solid on HAMS than HACS, as could be ascribed to the contribution of the micropores structure of HAMS. Similarly, HAMS adsorbed more serum proteins per gram solid than HACS when incubating in rat serum, and here the surfaces of both particles were almost completely covered by serum proteins, leading to almost the same protein adsorption amounts per unit area solid of HAMS and HACS. SDS-PAGE patterns proved that HA ceramic particles had different binding capacities for different serum proteins and was not highly dependent on the concentrations of the competing protein components present in rat serum. Western blotting analysis confirmed the enhanced adsorption of fibronectin and vitronectin on HAMS, indicating that HAMS might have better bioactivity than HACS. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2012.
Journal of Biomedical Materials Research Part B Applied Biomaterials 11/2011; · 2.15 Impact Factor
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04/2011; , ISBN: 978-953-307-191-6
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ABSTRACT: In order to enhance the ability of calcium phosphate-based biomaterials for bone defect repair, icariin (Ica), one natural product with ability of promoting osteoblasts differentiation in vitro and enhancing bone formation in vivo, was loaded into porous β-tricalcium phosphate ceramic (β-TCP) disks. The obtained Ica-loaded porous β-TCP ceramic (Ica/β-TCP) disks were characterized by SEM. The SEM photos indicated that the disks had porous structure and the surface morphology of the porous β-TCP ceramic (β-PTCP) disks had no obvious difference from the Ica/β-TCP disks. The Ica release curve of Ica/β-TCP disks showed a burst release during the first 1 day and the concentration of released Ica during the first 3 days had low cytotoxicity. The loading Ica in Ica/β-TCP disks hardly affected the attachment and morphology of Ros17/28 cells, however, the Ica/β-TCP disks were favorable to supporting the proliferation and differentiation of Ros17/28 cells better compared with the β-PTCP disks. There was plenty of bone-like apatite formed on the surface of Ica/β-TCP disks soaked in SBF solution for three days. After back intramuscular implantation of rats for three months, no obvious osteogenic evidence was detected in β-PTCP disks, but new bone formation was observed in Ica/β-TCP disks. Fibrous tissues and slight inflammatory reaction was also found in the Ica/β-TCP disks and β-TCP disks. Therefore, the loading Ica did not change the biocompatibility of β-TCP ceramic, but enhanced the bioactivity of β-TCP ceramic in vivo. The Ica/β-TCP ceramic had potential to be used for bone defect repair.
Journal of Materials Science Materials in Medicine 12/2010; 22(2):371-9. · 2.32 Impact Factor
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ABSTRACT: The biological performance of biomaterials is strongly influenced by their protein adsorption characteristics, which are related to the structures and properties of both the biomaterial and the protein. In the present study two groups of hydroxyapatite (HA) and biphasic calcium phosphate (BCP) ceramic powders were fabricated by different drying processes. The roles of the phase composition and microstructure of the powders in the adsorption of various model proteins were evaluated. The experimental results showed that BCP always had a higher ability to adsorb fibrinogen, insulin or type I collagen (Col-I) than HA. The microporosity and micropore size of the CaP particles also had a strong impact on their protein adsorption characteristics. HA and BCP particles with higher microporosities and/or more micropores >20 nm in diameter could adsorb more fibrinogen or insulin. However, amounts of adsorbed Col-I were largely unaffected by the microstructure of HA and BCP particles.
Acta biomaterialia 10/2009; 6(4):1536-41. · 3.98 Impact Factor
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ABSTRACT: Chondrogenic differentiation of mesenchymal stem cells (MSCs) relies on inductive media of chondrogenic environment. With proper design, a cellular microenvironment mimicking chondrogenic environment might be created to induce chondrogenesis of MSCs. In this study, bone marrow mesenchymal cells (BMSCs) were encapsulated in collagen-based hydrogel, and then enclosed in diffusion-chambers which allow the body fluid to permeate and preclude the host cells to invade. Then, the chamber with the hydrogel-BMSCs composite was implanted in the back of rabbits subcutaneously. The specimens in the chamber were harvested for histological, immunohistochemical, and RT-PCR analyses after 8 weeks. The results showed that cells with the characteristic of chondrocytes were homogenously distributed and the extracellular matrix (ECM) of cartilage has been secreted, indicating the chondrogenic differentiation of BMSCs. As control, nothing was obtained with only BMSCs. Moreover, the expression of collagen type II, indicator of cartilage ECM, was less in tissues with collagen-alginate-hydrogel (CAH) than that with collagen-hydrogel (CH). The results showed that both CH and CAH may induce the chondrogenesis and the induction is materials dependent. From in vitro experiments, TGF-beta is a necessary signal molecule for chondrogenesis, and it was suggested that the material may take in vivo growth factors to trigger chondrogenesis. From the studies, the chondrogenic induction of the hydrogel may be ascribed to that the hydrogel may provide a suitable environment and aggregate the signal molecule for chondrogenesis in vivo. The results would lend valuable reference in clinical for selection of appropriate scaffold for cartilage repair.
Journal of Biomedical Materials Research Part A 09/2009; 93(2):783-92. · 2.63 Impact Factor
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ABSTRACT: Protein adsorption affects the function of cells and determines the bioactivity of biomaterial implants. Surface structure and properties of materials determine the behavior of protein adsorption. In the present study, two biphasic calcium-phosphate ceramics (BCPs) with different surface structures were fabricated by pressing and H2O2 foaming methods. Their surface characteristics were analyzed and the in vitro and in vivo protein adsorption on them was investigated. Porous BCP showed higher ability to adsorb proteins, and transforming growth factor-beta1 (TGF-beta1) adsorption notably increased with increasing in vivo implantation time. The strong affinity of BCP to TGF-beta1 might provide important information for exploring the mechanism of the osteoinduction of calcium phosphates.
Acta biomaterialia 01/2009; 5(4):1311-8. · 3.98 Impact Factor
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ABSTRACT: 9-nitro-20(S)-camptothecin (9-NC) is a potent topoisomerase-I inhibitor, and it was applied for clinical trials in cancer treatment. However, the applications of 9-NC were limited by its poor solubility and instability. In order to overcome these disadvantages, 9-NC was encapsulated in amphiphilic copolymer micelles composed of methoxy poly(ethylene glycol)-b-poly(D,L-lactide) (mPEG-PDLLA, PELA). Three diblock copolymers with different PDLLA chain lengths were synthesized. The critical micelle concentration was varied from 10(-4) g L(-1) to 10(-2) g L(-1). The 9-NC loaded micelles were nanospheres with diameters ranging from 30 nm to 60 nm. The relationship between the composition of copolymers and the drug loading content was discussed. The encapsulation of micelles improved the solubility of 9-NC greatly. The solubility of 9-NC in micelle M1 was about 250 times higher than that of 9-NC in a phosphate buffer solution (PBS). The stability of 9-NC in micelles was also promoted. After being incubated in PBS for 160 min, 80% of 9-NC in micelles existed as an active lactone form, while 85% of 9-NC in PBS were transferred to an inactive carboxylate salt form. The release experiments were carried out in PBS and the results showed that the release processes were controllable.
Biomedical Materials 04/2008; 3(1):015013. · 2.16 Impact Factor
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ABSTRACT: Bone-like apatite on HA/TCP ceramics sintered at 1,100 degrees C (HT1) and 1,200 degrees C (HT2) could be obtained via immersing substrates into simulated body fluid (SBF) for 3 days. When MC3T3-E1 preosteoblastic cells cultured on the surface of the bone-like apatite for 3 days, SEM observations revealed cell membrane features with secreted crystals very similar to in vivo bone formation during intramembranous ossification with a direct bone apposition on the ceramics. According to semi-quantitative RT-PCR method, mRNA expressions of osteocalcin (marker of late-stage differentiation) and type 1 collagen were increased in cultures with HT1S and HT2S when compared to HT1 and HT2 after cultured for 6 days. The results indicated that bone-like apatite had the ability to support the growth of osteoblast-like cells in vitro and to promote osteoblast differentiation by stimulating the expression of major phenotypic markers. Taken together, our findings will be helpful in understanding the mechanism of osteoinductivity of calcium phosphate ceramics and in constructing more appropriate biomimetic substrate.
Journal of Materials Science Materials in Medicine 12/2007; 18(11):2237-41. · 2.32 Impact Factor
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ABSTRACT: The aim of this research is to study the effect of the controlled releasing character of the salmon calcitonin (S-CT) loaded injectable calcium phosphate cement (CPC) modified by adding organic phase, chitosan oligosaccharide (CO) and collagen polypeptide (CP). The uniform design was used to determine the basic formulation with suitable injectable time for clinical application, and then the changes of the physical characters, the controlled releasing character of the modified CPC along with the ratio of the organic phase were also evaluated in vitro. The surface morphous of the modified CPC been implanted in the abdominal cavity or soaked into the serum of rat was also observed by scanning electron microscope (SEM). The result shows that a suitable formulation of modified CPC could be got, and the injectable time is 12 min, the compressive strength is 12 MPa, and the final setting time is 40 min. Comparing with the CPC without organic phase, the releasing rate of S-CT would increase along with the increase of the organic phase after 7th day. Therefore, a novel S-CT loaded bioactive injectable CPC for treating osteoporosis induced bone defect was obtained, and the release of the containing S-CT was controlled easily through adjusting the ratio of CO and CP.
Journal of Materials Science Materials in Medicine 12/2007; 18(11):2225-31. · 2.32 Impact Factor
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ABSTRACT: Characterizations of hydroxyapatite (HA), biphasic calcium phosphate (BCP) and beta tricalcium phosphate (beta-TCP) ceramic particles were carried out using X-ray diffusion (XRD), Scanning electron micrograph (SEM), Particle Sizer and Zeta potential analyzer. Competitive adsorption of bovine serum albumin (BSA) and lysozyme (LSZ) on the three calcium phosphates were investigated by polyacrylamide gel electrophoresis (PAGE) method. The results showed that HA, BCP and beta-TCP ceramic particles with irregular shapes and similar size distributions all had negative surface net charges in pH7.4 phosphate buffered saline (PBS) solution and exhibited alike behaviors of BSA and LSZ adsorption. LSZ had higher affinity for calcium phosphate ceramics than BSA and its adsorption on them didn't be almost influenced by the increasing of BSA concentration in the solution. Electrostatic interaction played an important role on the competitive adsorption of BSA and LSZ on the surface of calcium phosphate ceramic particles.
Journal of Materials Science Materials in Medicine 12/2007; 18(11):2243-9. · 2.32 Impact Factor
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ABSTRACT: In this article, bioactive nanotitania ceramics with biomechanical compatibility was prepared by using an additive of hydroxyapatite or MgO as particle growth inhibitor. After sintering at 1000 degrees C, the particle size of nanotitania ceramics prepared by using HA as additive (HT) was much smaller than that prepared by using MgO as additive (MT). In simulated body fluid (SBF), HT could induce apatite formation in 4 days, while no apatite could be found on MT even after it was soaked in SBF for 14 days. After Ros17/28 osteoblasts were cultured on the materials for 1, 4, and 6 days, MTT results showed that the osteoblasts on the HT differentiated faster than that on the MT. Mechanical tests results showed that the bending and compressive strength of HT were 160 and 200 MPa, while those of MT were 70 and 88 MPa, respectively. These results demonstrated that it is suitable to prepare bioactive nanotitania ceramics, with biomechanical compatibility, by using HA as particle growth inhibitor.
Journal of Biomedical Materials Research Part A 11/2006; 79(1):210-5. · 2.63 Impact Factor
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ABSTRACT: In order to achieve biological sealing and resist mechanical damage of load-bearing percutaneous devices, Ti with excellent mechanical properties was anodic-oxidized to be endowed with bioactivity, with plasma-sprayed hydroxyapatite coated Ti as control. Similar to previous works, hydroxyapatite coating could bond tightly with living tissues, resulting in implant stability for whole implantation periods. Meanwhile, when anodic-oxidized bioactivated Ti was implanted percutaneously in vivo, it could induce a layer of calcium phosphate at the interface of tissues/implant. This layer of Ca-P not only induced the fibrous tissue or collagen ingrowth in its structure, but also improved the osteointegration between the bone and the implant. There was no significant biological response difference for the anodic-oxidized Ti and HA coated Ti at different implantation period with histological statistical analysis (p>0.05). Accordingly, suitable bioactivated modified surface of Ti by anodic-oxidized method could not only obtain the same results as the HA coating, but also might avoid some drawbacks of plasma-sprayed HA coatings to achieve biological sealing for a long period in vivo.
The International journal of artificial organs 06/2006; 29(6):630-8. · 1.86 Impact Factor
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ABSTRACT: The titanium surfaces containing calcium, phosphate ions and the carbonate apatite were characterized. The effect of surface chemistry on the initial rabbit osteoblast response on these surfaces was investigated. The cell count and alkaline phosphatase (ALP) specific activity assay were used for biochemical analyses. Scanning electron microscopy was used for morphology observation and in particular X-ray photoelectron spectroscopy (XPS) for surface chemistry characterization. The number of cells adhering to the apatite coating surface was the maximum, the number of cells on the surface containing calcium without phosphate ions was higher than that containing phosphate without calcium, and the number on the unmodified titanium surface was the least. The osteoblasts cultured on the apatite surface exhibited the highest ALP specific activity, next were the ones on the surface containing solely calcium, the lowest were on the unmodified titanium surface. On the substrate surfaces removed of adhered cells, the order of nitrogen amounts detected by XPS was consistent with ones of ALP specific activity and cell number, except for the unmodified titanium surface. For the substrate surfaces removed of adhered osteoblasts, XPS analysis showed that calcium and phosphorous amounts decreased during cell adhesion. After cell culture the Ca2p binding energy (BE) values for apatite coating and the surface containing solely calcium were similar to those of the two surfaces adsorbed bovine serum albumin (BSA). The P2p BE values for the surfaces containing phosphate ions, including the apatite coating and the surface containing solely phosphate ions, showed the same change. But after cell culture the decrease of the P2p BE value for the coating surface was larger than the one for the surface containing solely phosphate ions. Considering the bovine serum albumin adsorption on the same samples, these results indicated that calcium ions on titanium surfaces play a more important role than phosphate ions in initial interactions among culture medium, osteoblasts and titanium surfaces. On the apatite coating surface, calcium ions are active sites for osteoblast adhesion, while calcium and phosphate ions co-exist on titanium surfaces, the former promotes the osteoblast adhesion onto the phosphate sites on titanium surfaces. The cell adhesion was a complicated biological and chemical process relating to surface several elements similar to protein adsorption.
Biomaterials 09/2004; 25(17):3421-8. · 7.40 Impact Factor
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ABSTRACT: The aim of the present study is to demonstrate the newly formed tissue in calcium phosphate (Ca/P) ceramics after extra-osseous implantation by histological and immunohistochemistry (IHC) methods. Synthesis porous Ca/P ceramics without adding any growth factor and living cell were implanted in the dorsal muscle of dogs for 1 and 2 months. Undecalcified and decalcified sections were stained by hematoxylin and eosin (H&E), and IHC, respectively. The histological results showed the beginning of osteogenesis and angiogenesis after being implanted for 1 month and the obvious new bone formation after being implanted for 2 months. IHC were conducted via the avidin-biotin peroxidase complex (ABC) method and the primary antibody was collagen type I. IHC results indicated that collagen type I was expressed within osteoblast-like cells and newly formed bone-like tissue in Ca/P ceramics after 1 month, and in the mineralized matrix of newly formed bone and osteoblasts, some osteocytes and some lacunae after 2 months. No cartilage and chondrocytes were observed in the histological and IHC-stained sections. Evidence of intramembranous osteogenesis was confirmed.
Biomaterials 02/2004; 25(4):659-67. · 7.40 Impact Factor
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ABSTRACT: The relationship between surface characteristics of titanium and initial interactions of titanium-osteoblasts was investigated. Titanium plates were heat-treated in different oxidation atmospheres. The third passage rabbit osteoblasts were cultured on the titanium plates for 24h. After the heat-treatment, the crystal structure of the surface oxide films on titanium was identified using X-ray diffractometer and X-ray photoelectron spectroscopy (XPS). The surface roughness of titanium was measured with a profilometer. The surface energy was obtained by measurement of contact angles and calculation with Owens-Wendt-Kaeble's equation. The amount of surface hydroxyl (OH)(s) groups was examined using XPS. The change of binding energy of the some elements on the substrate surface suggested that the interactions between the cells and the titanium involved chemical reactions. The greater surface roughness, higher surface energy and more surface hydroxyl groups resulted in greater numbers of adhered osteoblasts and higher cell activity. Compared to the acidic hydroxyl (OH)(a) groups in (OH)(s) groups and the dispersion component of the total surface energy, the basic hydroxyl (OH)(b) groups and the polar component play more important roles in the osteoblast-titanium interaction.
Biomaterials 12/2003; 24(25):4663-70. · 7.40 Impact Factor
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ABSTRACT: Biological properties of titanium implant depend on its surface oxide film. In the present study, the surface oxide films on titanium were characterized and the relationship between the characterization and bioactivity of titanium was studied. The surface oxide films on titanium were obtained by heat-treatment in different oxidation atmospheres, such as air, oxygen and water vapor. The bioactivity of heat-treated titanium plates was investigated by immersion test in a supersaturated calcium phosphate solution. The surface roughness, energy morphology, chemical composition and crystal structure were used to characterize the titanium surfaces. The characterization was performed using profilometer, scanning electronic microscopy, ssesile drop method, X-ray photoelectron spectroscopy, common Bragg X-ray diffraction and sample tilting X-ray diffraction. Percentage of surface hydroxyl groups was determined by X-ray photoelectron spectroscopy analysis for titanium plates and density of surface hydroxyl groups was measured by chemical method for titanium powders. The results indicated that heat-treatment uniformly roughened the titanium surface and increased surface energy. After heat-treatment the surface titanium oxide was predominantly rutile TiO(2), and crystal planes in the rutile films preferentially orientated in (1 1 0) plane with the highest density of titanium ions. Heat-treatment increased the amount of surface hydroxyl groups on titanium. The different oxidation atmospheres resulted in different percentages of oxygen species in TiO(2), in physisorbed water and acidic hydroxyl groups, and in basic hydroxyl groups on the titanium surfaces. The immersion test in the supersaturated calcium phosphate solution showed that apatite spontaneously formed on to the rutile films. This revealed that rutile could be bioactivated. The analyses for the apatite coatings confirmed that the surface characterization of titanium has strong effect on bioactivity of titanium. The bioactivity of the rutile films on titanium was related not only to their surface basic hydroxyl groups, but also to acidic hydroxyl groups, and surface energy. Heat-treatment endowed titanium with bioactivity by increasing the amount of surface hydroxyl groups on titanium and its surface energy.
Journal of Materials Science Materials in Medicine 06/2002; 13(5):457-64. · 2.32 Impact Factor
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ABSTRACT: In previous investigations, a simple method, precalcification, was developed for bioactivating titanium. After a titanium sample was precalcified in a boiling saturated Ca(OH)(2) solution and then immersed in a calcium phosphate supersaturated solution, an apatite coating rapidly precipitated onto its surface. In the present study, heat-treatment in water vapor was carried out prior to precalcification. Heat-treatment in water vapor stimulated the chemical reaction between titanium, calcium, and phosphate. Coating properties were improved, and the bond strength of the coating to substrate was enhanced.
Journal of Biomedical Materials Research 02/2002; 59(1):12-7.
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ABSTRACT: Chemical treatments have been thought to be promised methods for improving bioactivity of titanium. In this work, the effect of precalcification with boiling saturated Ca(OH)2 solution on bioactivation of titanium was investigated. After precalcification and soaking in supersaturated Ca-P solution (SCP), calcium phosphate rapidly precipitated onto the surfaces of titanium, and after only three days an uniform apatite layer was found up to thickness of a few micrometers. The observation using scanning electron microscopy (SEM) showed that the coating was composed of a number of small crystal grains. The investigation by X-ray energy dispersion spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) indicated that the coating was Ca-deficient carbonate apatite. Based on the analyses for the surfaces and SCP, a mechanism of precipitation of apatite was proposed in thermal dynamics and kinetics.
Biomaterials 02/2002; 23(1):173-9. · 7.40 Impact Factor
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Journal of Materials Science Letters 12/2001; 21(2):153-155.
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ABSTRACT: The mechanism of apatite deposition on chemically treated Ti surfaces still is being studied. In this study, simulated body fluid, calcium aqueous solution, phosphate aqueous solution, and accelerated calcification solution are used as media to investigate the order of calcium and phosphate ion deposition on chemically treated Ti surfaces. The results of inductively coupled plasma spectra, scanning electron microscopy, and energy dispersive X-ray analysis show that calcium deposition is the prerequisite for phosphate ion deposition.
Journal of Biomedical Materials Research 12/1999; 47(2):213-9.
