Search

Mechanical strength and biocompatibility are issues of most concern for scaffolds in cartilage tissue engineering. Collagen modification is always used to strengthen scaffolds. There are mainly two ways for collagen modification: inclusion of reinforcing phase to form composites and chemical cross-linking. To explore an alternative approach, the collagen hydrogel modified by a reinforcement phase was compared with cross-linking. Collagen-alginate hydrogel (CAH) and collagen hydrogel cross-linked by genipin (CGH), which were different in modification methods, were chosen candidates. A comprehensive study was carried out on mechanical, structural and biological properties including swelling ratio measurement, in vitro degradation, AFM, mechanical test, thermogravimetric analysis, and in vitro cartilage tissue engineering. The results showed that mechanical strength of collagen was more enhanced for CGH than CAH, as evidenced by analysis of swelling ratio, in vitro degradation, AFM, mechanical test and thermostability. MTT and histological results showed that CGH was superior to CAH with less cytotoxicity and more chondrocytes distributed as well as more aggrecan secreted. With the increase in culture time, the cytotoxicity of cross-linker may be alleviated. CGH may provide a more favorable biomimetic environment for cartilage growth. All these indicated that selecting a cross-linker with a minimal cytotoxicity could be more promising for collagen modification, with improvements observed in both physical and biological properties. For reinforcement, it was required that the incorporated component should be equipped with better or equivalent properties compared with collagen. This study provided important implications to engineering collagen-based hydrogels for cartilage graft applications.

Bone tissue engineering (BTE) is approached via implantation of autogenous mesenchymal stem cells (MSCs), marrow cells, or platelet-rich plasma, etc. To the contrary, gene therapy combining with the bone marrow (BM) has not been often reported. This study was performed to investigate whether a modified BTE method, that is, the BM and a recombinant human bone morphogenetic protein-2 adenovirus (Ad.hBMP-2) gene administering in hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) ceramics could accelerate the healing of segmental defects in the rabbit radius. In our study, ceramics were immersed in the adenovirus overnight, and half an hour before surgery, autologous BM aspirates were thoroughly mixed with the ceramics; at the same time, a 15-mm radius defect was introduced in the bilateral forelimbs of all animals, after that, this defect was filled with the following: (1) Ad.hBMP-2 + HA/β-TCP + autologous BM (group 1); (2) HA/β-TCP + Ad.hBMP-2 (group 2); (3) HA/β-TCP alone (group 3); (4) an empty defect as a control (group 4). Histological observation and μ-CT analyses were performed on the specimens at weeks 2, 4, 8, and 12, respectively. In group 1, new bone was observed at week 4 and BM appeared at week 12, in groups 2 and 3, new bone was observed at week 8 and it was more mature at week 12, in contrast, the defect was not bridged in group 4 at week 12. The new bone area percentage in group 1 was significantly higher than that in groups 2 and 3. Our study indicated that BM combined with hBMP-2 adenovirus and porous ceramics could significantly increase the amount of newly formed bone. And this modified BTE method thus might have potentials in future clinical application.

In the present study, three types of tetracalcium phosphate (TTCP) were prepared by solid-solid reaction or co-precipitation method and by different cooling modes. The effect of TTCP on the performance of calcium phosphate cement (CPC) was investigated. The result showed that the characteristic of TTCP varied with preparation method and played an important role in CPC performance. A solid-solid reacted TTCP yielded smaller particle size and resulted in bad workability and mechanical strength of CPC. The fast cooling of sintering TTCP by liquid nitrogen could avoid the decomposition of TTCP and make pure TTCP. TTCP prepared by wet-precipitation could improve performance of CPC and was promising to optimization of CPC.

Collagen has been widely used in biomedical field, such as scaffolds for tissue engineering. However, the rapidly biodegradation and weak mechanical strength of collagen limited its application. In this study, an insoluble collagen extracted from cattle hide was designed as scaffold to act as a three-dimensional substrate for tissue engineering. The received insoluble collagen sponge was analyzed by scanning electron microscopy, infrared spectroscopy, mechanical testing and thermogravimetric-differential thermal analysis. In addition, the degradation was performed in vitro using collagenase. The results showed that the insoluble collagen had the same triple helical domain as acid-soluble collagen, while the compression strength was greatly improved and the degradation rate was reduced. The insoluble collagen sponge with good stability should be promising in tissue engineering scaffold applications.

With good biocompatibility, collagen is often used in cartilage tissue engineering. Collagen/alginate composite was hoped to improve the poor mechanical property of pure collagen but the biocompatibity was decreased. In this study, hydroxyapatite (HA) particles were used to get collagen/alginate/HA (CAHA) composite film to enhance the bioactivity properties. The bioactivity of the composite was investigated by in vitro co-culture with chondrocytes. During the 6-day cell culture in vitro, the composite showed a significant improvement in promoting proliferation and maintaining morphology/phenotype of the chondrocytes over collagen/alginate composite by MTT, SEM, fluorescent and immunohistochemical assays. Cytocompatibility and cytoviablility of CAHA even come up to that of collagen film alone. The results indicated that the composite film may provide an appropriate environment for the proliferation and maintaining the morphology and phenotype of chondrocytes and have a potential clinical application in the cartilage tissue engineering field.

It is well known that the cellular responses are related with both physical and chemical characteristics of substrate, including surface topography. In the present study，the effect of surface topography of hydroxyapatite (HA) on rat bone marrow cell (rBMCs) response was investigated. HA disc-shaped pellets with various topography were manufactured by single-axis pressing methods. The rBMCs responses on materials including cell morphology and proliferation were evaluated by SEM and MTT methods respectively, and the differentiation potential was assessed by total protein content and alkaline phosphatase (ALP) activity testing. The results showed that the cell porliferation was higher on HA surfaces with macropore structure, while ALP activity was lower. No significant difference in the cellular responses on the pore distribution and orietation was observed. However, the pore structure had a potential to guide cell orientation by gathering the cells inside the pores rather than on the ridges. Since ALP served as an indicator of early osteoblast differentiation, in this study its higher expression on HA surface with micropores suggested that surface microtopograhy exhibited an important effect on early osteoblast differentiation process.

L9 (34) orthogonal array design was employed to optimize experimental conditions for the preparation of the composite using in situ synthesis method and to analyze the relationships between experimental parameters and mechanical property of the composites. Bending strength of the composite was considered as a target property of the composites. Hydroxyapatite content in the composite, synthesis temperature and pH were chosen as main parameters. As a result of this study, bending strength of the composite appeared in peak with the increase of the hydroxyapatite content of the composites and synthesis pH, while with the increase of temperature, bending strength decreased. Optimum experimental conditions for the synthesis of the composites with higher bending strength were determined. The bending strength of the composites was 90 MPa at the optimal synthesis conditions.

Stress concentration is one of the main mechanical problems leading to the failure of clinical application for osteointegrated implant of percutaneous osteointegrated prosthesis, which is especially marked for higher amputated leg prosthesis. Traditionally design was composed of only the distal part. To improve the biomechanical safety, a new design with the lag part similar to the lag screw was introduced. Based on CT scan data, relatively accurate model of femur for finite element analysis (FEA) were obtained. The FEA results with the new implant demonstrated that compared to traditional design, the declination of bone stress peak ranged from 15.68% to 28.67%, perpendicular deformation from 34.73% to 72.16%, and maximal stress of implant from 14.51% to 23.36% with the increasing of loads from 3750N to 2000N. So the new design of osteointegrated implant would be more secure mechanically, in the case of higher amputated leg attachment.

The effect of different hydroxyapatite particles on malignant melanoma cell was evaluated in vitro. Two kinds of short-rode hydroxyapatite particles, which range from 0.4 to 1 um and 0.5 to 1.5 um, were co-cultured with human’s malignant melanoma (MM) cell line A375 for 24, 48, and 72 h. Both of these HA particles showed the same cell proliferation rate as blank control, and there was no statistically significant correlation of matrix metalloproteinases-2 (MMP-2) expression to the HA particle size. SEM images showed that the membrane of tumor cell, which co-cultured with HA particles, was shrunk. The cell superficial sentus was reduced and small size particle’ effect was more obvious. Since immunohistochemistry and SEM data are qualitative technique, further precise methods might bring more information about the effect of HA particles on tumor behaviors. But this study can provide the bio-security test of micrometer HA particles in transplantation after tumor excision.

Linear and Star-shaped PCL-b-PEG copolymers were synthesized through a two step process, including the first step to synthesize the star-shaped PCL through ring-opening polymerization of Γ-caprolactone initiated from multi-functional alcohol under the existence of tin(II) ethylhexanoate [Sn(Oct)2] catalyst, and the following step to couple the obtained star-shaped PCL with PEG segments using bi-functional linker. The structure of the polymers was confirmed by IR, NMR, GPC, et al. The aggregation behaviors of the star-shape copolymers were compared with that of the linear block copolymer with corresponding molecular weight of each arm, and the influences of structure factors were discussed.

As a natural biomaterial, collagen especially pepsin-solubilized collagen (type I) has been used widely in biomedical fields due to its excellent biocompatibility. In this preliminary study, we investigate the effect of some inorganic ions which are frequently utilized in the preparation of collagen on the morphology and crystallinity of fibrils. The scanning electron microscope and x-ray diffraction were applied to analyze the morphology and the crystallization of the reconstituted collagen fibrils, respectively. Although further studies are needed, these initial results indicate that by controlling the self-assembly conditions of collagen molecules, we may achieve the desired properties of fibrillar collagen products.

The study is aimed at to explore the construction of bone graft with calcium phosphate ceramics implanted in subcutaneous fat, and provide applying technique for the in vivo bone tissue engineering. Forty-eight calcium phosphate ceramics (Ca-P ceramics) columns with Φ 5×8 mm were prepared, and eight dogs were used in this experiment. Six samples were implanted in each dog. Three in dorsal muscles while the others in subcutaneous fat. The specimens were harvested at 4, 6, 12 and 24 weeks post operation, for gross observation, SPECT and histological studies. The osseous or osteoid tissue formation at different times in the two non-osseous sites was compared and the new bone grafts in subcutaneous fat and intramuscular implantations were evaluated. The results demonstrated that the bone graft could be constructed not only in muscle but also in subcutaneous fat. Compared to that in muscle, constructing bone graft in subcutaneous fat could have brighter prospect to clinical application.

The bioactivities of titanium oxide film on titanium surface received from different chemical treatment methods were studied in SBF in vitro and mechanically and histologically investigated in vivo. Three groups of titanium specimens were prepared: untreated titanium(S), acid-alkali treated titanium (H), and acid-alkali and heat-treated titanium(X). The oxide film of X surface resulted in more apatite formation and significantly higher strength of the interface between the samples and bone than those of the other titanium groups. The surface of the acid-alkali treated titanium and that further treated by heat treatment had higher bioactivity and stronger bone-bonding ability.

Zirconia toughened alumina (ZTA) nano-ceramic composite has excellent mechanical properties. Preparation of ZTA was conducted in the present study with an attempt to use as dental ceramics. On the basis of the study on the effect of the die pressing forces on the density and Vickers hardness of the sintered ZTA, additive aids TiO2, MgO, SiO2 and CaO were introduced in order to reduce the sintering temperature of ZTA nano-ceramic composite with good densification. Scanning electron microscopy was employed to evaluate the microstructural morphology. Phase composition was confirmed by using X-ray diffraction. Thermal analysis was further used to study the structural change. The results indicated that the ZTA product sintered at 1450°C had dense and uniform microstructure due to the combinative effect of additive aids, including the formation of an intermediate compound. This study suggested that such ZTA nano-ceramics had potential applications as dental prosthetic materials.

The achievement of biological sealing is determined by the quality of the skin attachment on the surface of the percutaneous implant in the area where the implant penetrates the skin. It has been known that certain surface features of the implants can significantly influence the interactions between cells and substrate. In this study, titanium plates were bioactivated through anode-oxidization firstly, and then cultured with human epithelium cells for 72h. Untreated Ti plates were used as control. After the samples were dehydrated, the morphology of the cultured epithelium cells was tested with Scanning electron microscopy (SEM). The surfaces of control group did not enhance epithelium cell attachment and growth, while the bioactivated microporous surface of anode-oxidized group would be beneficial to induce the formation of the pseudopod of epithelium cell, and then interlock the human epithelium cells through the pseudopod, which imply that the surface modification method of anode oxidization may be one of the most effective methods to resolve the biological sealing.

Nano-titania ceramics is a potential biomaterial for orthopaedic application. In our previous studies, a bioactive nano-titania ceramics was prepared by using alkali-heat treatment. In this paper, hydroxyapatite was used as a grain growth inhibitor additive to get nano-titania ceramics with different grain size, and the effect of grain size on the bioactivity was studied in vitro.

This study was carried out to investigate the effect of acid-alkali treatment and alkaliheat treatment on the push-out strength and tissue response of the porous titanium in vivo. Porous titanium with different treatment was implanted in dog bony site for 2 months and 5 months and the push-out strength was tested. At 2 months, the mean push-out strengths of the acid-alkali treated and alkali-heat treated porous titanium were 11.3 and 15 MPa, respectively. At 5 months, the values reached 29.8 and 35 MPa, respectively. Histological observation showed a close contact between implants and bone, and more bone tissue filled inside the pores of porous titanium increasing with implantation time. The results indicated higher bonding strength between bone and porous titanium in alkali-heat treated samples. Therefore, alkali-heat treatment can provide porous titanium implants with better fixation as a bone substitute for clinical use under load-bearing conditions.

After the Tantalum metal was subjected to the anodic oxidation at suitable voltage in 2M H2SO4 solution, tantalum oxide with rhombic or amorphous structure formed on the metal surface. The Oxide showed apatite formation ability in simulative body fluid at 6d. It meant the anodic oxidation treatment ia an effective method to accelerate the bioactivity of tantalum metal.

The aim of this study is to reveal the response of highly malignant tumor cell to different hydroxyapatite particles. Human’s highly malignant melanoma (MM) cell line A875 is exposed to two kinds of short-rod hydroxyapatite particles. After incubated with hydroxyapatite particles at different time points, the effects of MM cell morphology, proliferation and invasion are evaluated by SEM, MTT, and MMP2 protein immunohistochemistry respectively. SEM figures show that superficial sentus of MM cells reduce; more secretion is found; and smaller HA particles have more obvious effect. At the same time, no statistic difference can be found in MTT assay and MMP2 expression measurement. This study also provides useful information about biosecurity of hydroxyapatite ceramic in transplantation after highly malignant tumor excision.

Porous tricalcium phosphate ceramics were immersed in static and dynamic revised simulated body fluid (RSBF) at 37°C. Morphology, composition and phase of precipitates on TCP were identified by SEM, FTIR and TEM methods. FTIR and TEM results indicated the deposits on the inner pore walls of TCP were OCP, and SEM results implied that the deposited way of precipitates in static system was different from that in dynamic system.

Circumambient tissues infection is a common complication of implanted percutaneous and permucosal devices (PDs). In order to counter the infection, the titanium abutment surface of PDs was modified to achieve antibacterial and antiadhesive activities. Titanium discs were immersed into hydroxyapatite (HA) sol containing different mass fraction silver and thermally treated to get modified discs. The modified discs become more smoother and get crystal HA coatings containing silver. Silver ion release test demonstrate the release rate is very slow and the coating is a reservoir of silver. The antibacterial and antiadhesive effects of modified discs on Staphylococcus aureus and Porphyromonas gingivalis were evaluated. Modified titanium discs with silver significantly inhibited the growth of these two bacteria compared with both the polished titanium discs control and modified titanium discs without silver. And titanium discs modified with more silver have higher antibacterial activity and less bacteria adhesion in this study. These findings indicate that titanium surface modified with the HA sol-gel containing silver could achieve antibacterial and antiadhesive activities, and the abutments of PDs are promising to be modified by this method.

In order to prepare nano-hydroxyapatite/poly(lactide) (n-HA/PLA) composite with good interfacial interaction, some groups which could bind with Ca ions in HA crystals need to be introduced onto PLA surface. Poly(α-methacrylic acid) (PMAA) was grafted on the PLA surfaces via photooxidization and subsequent UV induced polymerization. Suspension of PMAA-PLA microparticles with an average size as 133.1nm was prepared with solvent evaporation technique. Then utilizing the action of template manipulating of PMAA-PLA microparticles, n-HA/PLA composite were synthesized. Zeta potentials measurement and SEM indicated that there were good interfacial interactions between two phases of n-HA/PLA composite. The results of cell viability confirmed that n-HA/PLA composite possessed good cytocompatibility, so the n-HA/PLA composite scaffold obtained by electrospun technology might be used as bone tissue engineering scaffold.

Poly(vinyl alcohol) (PVA) was introduced during in situ synthesis of hydroxyapatite (HA) in neutral collagen (COL) solution and final PVA-COL-HA nanohybrids were achieved via sequential steps including gelation by fibrillogenesis, freezing-thawing physical crosslinking, removal of unreacted residues and dehydration. This method is expected to endow the pure PVA with good bioactivity and meanwhile the presence of elastic PVA would improve the properties of COL-HA composites. The phase, microstructure and possible molecular interactions of the achieved PVA-COL-HA nanohybrids were analyzed by using X-ray diffraction, Fourier transform infra-red spectroscopy and scanning electron microscopy. The results indicate that the inorganic phase is poorly crystallized apatite with a nanometer size due to the confinement of organic macromolecules which forms a network structure.

In order to develop a bone-filling material with osteoinductive potential, a composite micorspheres of collagen molecules and biphasic calcium phosphate (BCP) was prepared by utilizing emulsion polymerization and the intrinsic self-assembly of collagen. The prepared microspheres were analyzed by granularity test, scanning electron microscopy (SEM), infrared spectra (IR) and enzymatic digestion experiment. The results showed that the collagen matrix of fibrils was reconstituted in the droplets, and the native triple-helix structure of collagen was still maintained. The study provides an effective way to prepare microspheres of collagen and BCP composite.

Collagenous molecule was successfully immobilized to hydroxyapatite (HA) surface through a molecular bridge (2-Hydroxyethyl acrylate, HEMA) that was grafted to the surface with covalent bond by gamma irradiation. Hydroxyapatite modified by atelocollagen had been characterized by several surface sensitive techniques, such as FT-IR, SEM, XPS. The investigations showed that the collagen, a bioactive macromolecule, was immobilized on the HA surface through covalent bond.

Porous HA/TCP bioceramics were immersed in pure dog serum to observe apatite formation. Deposited crystals were examined using SEM. Results showed that beamed sheet-like crystals formed on the surface of ceramics granules, and after postponement immersion time, crystals extended and became bigger. EDS and IR results suggested formed crystals were defect-calcium type carbonated hydroxyapatite. HRTEM photograph suggested formation process of new-formed crystals from non-crystal to crystal in serum. Directional organisms acted maybe as a template in process of crystals formation, so new crystals developed along certain direction.

Protein adsorption is driven by various interactions. The contribution of surface charge to bovine serum albumin (BSA) adsorption on hydroxyapatite (HA) ceramic was investigated by adjusting the liquid environment in which the solid particles dispersed. Zeta potentials of HA and the adsorption of BSA on the surface were tested as a function of pH, ionic strength, Ca2+ and PO4 3- concentrations in the aqueous solutions, and the results showed that both of them were greatly affected by those experimental variations. Besides, the amount of adsorbed BSA was related to the variation of zeta potential of HA, as could be well understood in terms of electrostatic interactions.

Collagen (Col) and chitosan (Chi) are both natural polymers and have received extensive investigation in recent years in the field of tissue engineering, but there are few reports on the introduction of hydroxyapatite (HA) into the Col-Ch system. In this study, based on the miscibility of these two polymers under proper condition, hydroxyapatite (HA) was synthesis in the Col-Chi system by in-situ co-precipitate method to give rise to a novel nanocomposite. The structural characterization of such Col-Ch-HA nano-materials was carried out by using FT-IR, XRD, SEM and TGA analyses with main components and Col-Chi samples used for comparison. It was found that there exist interactions between Col and Chi molecules. The nucleation and growth of inorganic phase occurs in the Col-Chi system and final products are uniform dispersion of nano-sized HA in the Col-Chi network without obvious phase separation. This novel nanocomposite would be a promising material for bone tissue engineering.

Synthesis of hydroxyapatite (HA) in organic solutions has received extensive attention in recent years with an attempt to obtain HA of a nanometer level. In this preliminary study, we demonstrated that organic-HA nanocomposites could also be achieved with one step method via in situ mineralization and subsequent crosslinking of organic species. This design was realized through in situ synthesis of hydroxyapatite in poly(vinyl alcohol) and acrylic acid aqueous solution as an organic template. The aforementioned organic-inorganic nanocomposites were analyzed by using X-ray diffraction, Fourier-transform infrared spectroscopy, scanning electrical microscopy, thermal analysis. The comparative structural measurements were also conducted with the synthesized HA with absence of the organic template. The results indicated that the existence of organic species effectively inhibits the growth of calcium phosphate and that relatively pure HA can be obtained in sintered composite products. The present study provides a direct and versatile route for fabrication of nanocomposite biomaterials.

Hydroxyapatite (HA) ceramics is one of the most widely used bone substitute in clinics. Limited information is available concerning how HA ceramics may affect osteoprotegerin (OPG) and RANKL expression. In this study, osteoblastic-like SaOS-2 cells were grown on HA ceramics sintered at different temperature for 3 and 6 days, RANKL and OPG mRNA expression were analyzed with quantitative in situ hybridization (QISH) technique. Result showed that SaOS-2 grown on HA ceramics sintered at 800°C expressed higher RANKL mRNA than on other two HA ceramics after 3 days’ culture. No significant difference in OPG expression on different surfaces was detected after 3 days and 6 days culture. This result suggests that HA sintered at low temperature tend to induce more bone remodeling after implantation.

Hydroxyapatite/collagen composites were prepared in-situ synthesis. The composites were finally achieved by dehydration including air-drying and freeze-drying methods. FTIR, XPS and DSC were employed to investigate the composites dehydrated by two methods. The air-dried composites had better mechanical properties than those of the composites dried by freeze drying. Air-drying of the composite induced more bond formation and crosslink between collagen fibers and HA crystals compared with freeze-drying of the composite, as indicated by the shifting of amide A and I bands to the lower wavenumber and by the changes in the binding energy of O1s, Ca2p, and P2p, leading to the increase of the peak temperature of the composites. Collagen crosslink and bond formation in the air-dried composites were key factors to increase the bending strength of the composites. The results of this study confirm that in situ synthesis and air-dry method are effective ways to obtain nanoHA/COL composites with high mechanical properties.

The purpose of this study was to explore the feasibility of repairing massive bone defect with in vivo tissue engineering(TE) bone, and to provide experimental evidence for the application of in vivo TE bone into clinic in the future. Six calcium phosphate ceramics (Ca-P ceramics) columns were prepared, and then immersed in dynamic revised simulated body fluid (RSBF). 72 hours later, the bone-like apatite was formed on the surface and pore walls of ceramics. Three dogs were used in this study. Two ceramic columns were implanted bilaterally in the femoral muscles of each dog to construct living bone graft of in vivo TE bone. 6 weeks after implantation, they were transplanted to the box-like bone defects sites created in bilateral mandible of the same animals. The dogs were sacrificed at 8, 12 week after operation respectively. Samples were harvested for gross observation, X-ray examination, tetracycline fluorescence labeling, SPECT and histological observation. These results demonstrated that as a living bone graft, in vivo TE bone participated in the bone metabolism of host, and integrated with the host bone. It is feasible to reconstruct box-like bone defect of mandible with the in vivo TE bone.

The aim of this study is to prepare nano hydroxyapatite powder. Hydroxyapatite powder was prepared via co-precipitated method with the addition of citric acid at pH 9-11 in ambient environment. The precipitates were aged for 24hs, and then milled into powder after washed and dried. The particle morphology and particle size of as prepared HA powders were characterized. The results showed that hydroxyapatite powder with width of 10-30nm and length of 30-100nm was prepared by wet co-precipitation.

In this study, bovine serum albumin protein (BSA) was introduced to investigate the co-precipitation process of calcium phosphate and BSA on bioactivated Ti. Commercially pure titanium were bioactivated firstly, and then immersed in a highly supersaturated stable calcium phosphate (Ca-P) solution at three different conditions. The samples designated as Ti-C, Ti-C-CB, and Ti-C-B for control. The samples were evaluated by SEM with EDX, XRD and XPS. The co-precipitation of BSA protein and Ca-P influenced the morphology of the crystals of Ti-C-CB significantly. In terms of the immersion in the Ca-P solution containing BSA, the co-precipitation of Ca-P with BSA on the surface of Ti-C-CB was a chemical process rather than simple physical adsorption, which was most possibly achieved by the linkage of –COO− groups to Ca-P. Such coprecipitated interaction led to the formation of a tight, dense and uniform Ca-P coating.

In this study, three kinds of photo-reactive polymers, namely positively charged azidophenyl-derived polyallylamine (AzPhPAAm), negatively charged azidophenyl-derived poly(acrylic acid) (AzPhPAAc) and neutral azidophenyl-derived poly(ethylene glycol) (AzPhPEG), were synthesized by introduction of photo-reactive group (-N3) into polyallylamine, poly(acrylic acid) and poly(ethylene glycol), respectively. The structure of the azidophenyl-derived polymers was confirmed by 1H-NMR measurement. All of the photo-reactive polymers can be pattern-grafted on the surface of cells culture plate, which was confirmed by the optical microscopy observation. The contact angle decreased after surface modification by the photo-reactive polymers. Human mesenchymal stem cells (MSC) cultured on the modified surfaces showed different morphology. The cells adhered and spread more on the PAAm- and PAAc-grafted surfaces than on the PEG-grafted surface. The pellets formed on PAAm- and PEG-grafted surfaces in condrogenic differentiation medium were positively stained by safranin O/ fast green. But the pellet formed on PAAc-grafted surface did not show obviously positive staining for safranin O/ fast green. These results indicate that the PAAm- and PEG-grafted surfaces promoted chondrogenic differentiation of MSC.

This study investigated the utility of genetically modified cell line for fast and non-destructive cytotoxicity evaluation of biomaterials. The EGFP(enhanced green fluorescence protein)-expressing plasmid pcDNA-EGFP was constructed, and electroporated into ROS 17/28 osteoblastic cells to generate an EGFP-labeled stable cell line, ROS-EGFP. This genetically modified cell line provided two unique opportunities to qualitative and semi-quantitative evaluation of cell growth on biomaterials without destruction of samples. Firstly, utilizing the fluorescence of EGFP expressed in the cells, the viability state of cells on biomaterials was evaluated using a fluorescent light microscope. Secondly, the proliferation of cells on biomaterials, which was identified by MTT assay，was demonstrated according to the microscopically counted fluorescent cell numbers. From the results, it could be concluded that the ROS-EGFP cell line was an effective tool to trace the fate of cells on biomaterials and to evaluate the biocompatibility of biomaterials to cell growth in vitro.

The purpose of this study was to develop a feasible approach for repairing periodontal bone defects with the in vivo tissue engineering bone incorporated with bioabsorbable PLA membrane and to provide evidences for the clinical application. Osteoinductive HA/β-TCP sintered at 1100°C were implanted in the femur medial muscles in the hind legs of three dogs. Four weeks after implantation, the in vivo tissue engineering (TE) bone was explanted. Meanwhile, artificial periodontal bone defects of 8mm×6mm were performed on the buccal side of 4th premolar and 1st molar of mandible bilaterally, with the exposure of dental roots. The defects were treated as follows: (1) in vivo TE bone and PLA membrane; (2) HA/β-TCP ceramics and PLA membrane; (3) PLA membrane only; (4) empty control. At the 2, 4, 8 weeks post-transplantation, the dogs were sacrificed. The specimen were harvested and evaluated by gross inspection, dental radiography, SPECT (99mTC-MDP) and histological observation by MPIA2500. The results showed that more mature osseointegration was found in the group 1. We presumed that the in vivo TE bone graft could enhance the reparation of periodontal bone defects.

The mRNA expression of Cbfa1 and osteocalcin gene induced by calcium phosphate ceramics (Ca/P) were quantitative analyzed according to real-time RT-PCR method in this work. C2C12 cells were co-culture with four kinds of porous Ca/P ceramics for 2 and 5d without adding other growth factors. The four kinds of Ca/P ceramics were pure hydroxyapatite (HA) sintered at 1250°C and HA/TCP with a ratio of 60/40 sintered at 1100°C (HT1), 1200°C (HT2) and 1250°C (HT3) respectively. Real-time RT-PCR analysis found the Ca/P ceramics induced positive expression of Cbfa1 and osteocalcin in C2C12 cells, After 5 days culture, Cbfa1 and osteocalcin showed obvious higher expression compared with that in 2 days. Cbfa1 and osteocalcin expression in BCP was much higher than HA, and the expression level of osteocalcin was HT1>HT2>HT3>HA. Our results showed that Ca/P ceramics alone were sufficient to induce C2C12 cells to osteoblastic differentiation and the sinter temperature and phase composition of Ca/P ceramics could affect their osteoindctive capacity significantly.

The aim of this study was to optimize sintering aids and processing parameters for zirconia toughened alumina (ZTA) nano-ceramics with optimal properties as dental ceramics. Additive agents (TiO2, MgO, SiO2), dry pressing forces and sintering temperatures are very important for preparation of structural ceramics. In the present study, density and Vickers hardness measurement, SEM, and XRD analysis were employed to investigate the effects of these on the final products. The relevant results showed that higher pressing force led to an increase in the density and Vickers hardness of the sintered ceramics. Among the four groups of sintered nanoceramics with different ratios of alumina and zirconia, and combinations of additive agents, the best densification was achieved in the sample from Group 2# containing 1.05wt% MgO after sintered at 1450°C, as indicated by SEM observation. XRD analysis confirmed the formation of MgAl2O4 as an intermediate compound. The existence of TiO2 in the additive agents was in favor of the acquisition of high densification. The measured values for the density and Vickers hardness indicated that the sintered ZTA nano-ceramics would be a potential material for dental prosthetic applications.

Serum immersion, a new method of pre-treated HA ceramics, has been compared with SBF immersion and untreated HA particles in cell culture condition. MTT assay indicates that serum-immersion lessens particles cytotoxity of HA particles in cultured L929 mouse fibroblast cell (p

In vitro method has often been used in the biodegradation/bioactivity evaluation of bioactive ceramics for its convenience and saving in time and outlay. The simulated body fluid (SBF) suggested by Kokubo was a good simulation of the osteoproduction environment in osseous tissue and has been proved to be a good method to study the bioactivity of biomaterials and the mechanism of bone bonding. But SBF is not a suitable method to research the osteoinduction of biomaterials. The results from SBF were not consistent with that from in vivo in muscle. The local ion concentration is the key factors to affect the nucleation and growth of apatite. In muscle the effect of body fluid flowing on local ion concentration cannot be ignored. A dynamic SBF suggested by these authors of this paper not only simulated the ion concentration of body fluid, but also simulated the effect of body fluid flowing on the local ion concentration near the surface or in biomaterials in muscle. The results from the dynamic SBF were in good agreement with that of the implantation experiments in muscle. The results from dynamic SBF showed that apatite only formed on the walls of macropores of the porous CaP, no apatite formed on the surface of both dense and porous CaP. The new bone only formed on the walls of macropores of porous CaP implanted in muscles, no apatite or osseous tissue could be found on the surfaces of both porous and dense CaP. The dynamic SBF preferably simulated the osteoinduction environment in non-osseous tissue and can be used in osteoinductivity evaluation of bioceramics.

Bonelike apatite coating on metal implants is an effective method to enhance bioactive properties of the metal surface. In the present study, the effectiveness of cathode deposition (CD) coating methods was investigated. And biomimetic deposition (BD) was also investigated as the parallel. The revised simulated body fluid (R-SBF) was chosen as an electrolyte and simulated body fluid. Both deposition methods could produce bonelike apatite coating on rough surfaces of the titanium. The uniformity of the CD coatings were better than the BD coatings’. And CD method was less sensitive to the condition of the titanium plate surfaces and much faster in the coating deposition. The spectra of FTIR showed that the characteristic peaks of CO3 2- apeared on the surface of ceramics, combined with spectra of XRD and SEM, it was concluded that a sertain thickness of bonelike apatite coating could be formed on the acided treatment (AT) titanium plate surfaces by CD method.

Calcium phosphate cement (CPC) has many advantages and is frequently used as a carrier of antibiotic and bone growth factors. Salmon calcitonin(S-CT) is effective in treating osteoporosis. Due to its potential of promoting bone cell proliferation, S-CT was combined with injectable CPC to accelerate the restoring of the bone defect induced by osteoporosis and the composite can be used in minimal invasive surgery. Uniform design was used to optimize the prescription, and the effects of pore-maker and S-CT on the characters of CPC were studied. The in vitro releasing of S-CT from the optimum CPC in deionized water, SBF and rabbit serum was studied with HPLC, respectively. The results showed that the contents of HA and citric acid-NaHCO3 affect the physical characters of CPC sharply, but S-CT has little effect on it. The releasing of S-CT in three different medium follow Hugichi equation, but the speed is slower in rabbit serum than in deionized water and SBF. S-CT carried porous injectable CPC composite possesses the basic performance for clinical needs, and it is promising to be used in osteoporosis induced bone defect and accelerate bone repair.

An organic/inorganic composite hydrogel route was used to prepare collagen-calcium phosphate hybrids with high mechanical strengths, via in-situ mineral synthesis during collagen fibrillogenesis followed by dehydration. An array of characterization techniques including X-ray diffraction and Fourier transform infrared spectroscopy analyses confirmed that the final products are analogous to natural bone. A three-point bending strength of 70 MPa, much higher than the values reported in the literature, was recorded in the present case, due to the three dimensional network structure achieved between inorganic and organic phases. This innovative method provides an efficient route to produce bone grafts with the desirable mechanical properties which are dependent upon the actual inorganic/organic ratio and water content.

A composite hydrogel with interpenetrating network structure was prepared via in-situ synthesis of calcium phosphates during the physical-chemical crosslinking of poly(vinyl alcohol) (PVA) and poly(acrylic acid) (PAA). The hydrogel water content was tested. Fourier transform infrared absorption spectrum (FT-IR), X-ray diffraction and scanning electron microscopy were employed to evaluate the characteristics of the composite hydrogel. The results showed that the composite hydrogel had high water content and that the inorganic phase was poorly crystalline calcium phosphates. FT-IR confirmed that the interpenetrating network structure was formed between PVA and PAA. The chemical interactions between inorganic and organic phases were further investigated and discussed. The composite hydrogel with an interpenetrating network achieved using the present novel method could be a promising material for tissue engineering.

Good interfacial interaction is crucial for preparation of inorganic-organic materials at a nanometer level. Poly(α-methacrylic acid) (PMAA) was grafted on the PLA surfaces via photooxidization and subsequent UV induced polymerization in an attempt to synthesize nano-hydroxyapatite/poly(lactide) (n-HA/PLA) composites. Grafting of PMAA on the PLA surface was confirmed using FTIR analysis and the size distribution measurement of the grafted-PLA (g-PLA) particles. n-HA/g-PLA composites were in situ synthesized via dropwise addition of Ca2+- and g-PLA containing solution to PO4 3-- solution. The prepared composites were characterized by FTIR, XRD，SEM and TEM means. Analytical results indicated that the g-PLA acts as a template to manipulate the nucleation and growth of n-HA crystals and thereby to control the morphology, size and anisotropy of n-HA crystals and their distribution over the organic phase. Chemical linkages and/or interfacial interactions between the n-HA and the g-PLA in the n-HA/g-PLA composite were further discussed.

The electrochemical cathode bioactivating titanium with additive of citrate was reported. After the treatment, there was a uniform dense layer with very tiny granules on the surface of the titanium, this layer was composed of amorphous calcium phosphate-citrate (ACP-CIT) with a thickness of about 150nm. After immersion in saturated calcium phosphate solution, the morphology at different time points of the early stage was quite different. The changes of morphology showed that the citrate in amorphous calcium phosphate modulated the crystal growth.

Studying on the interaction between proteins and calcium phosphate implants is one of the basic subjects in biomaterials science and engineering. In this work, zeta potentials and contact angles of hydroxyapatite (HA) and biphasic calcium phosphate (BCP) ceramics were measured, and the adsorption behaviors of bovine serum albumin (BSA) on the surfaces were investigated. The adsorption isotherms of BSA on both ceramics follow the Langmuir type, however, BCP shows higher BSA adsorption ability. The differences of contact angles and zeta potentials in HA and BCP are the key factors to determine their ability to bind BSA molecules.

The purpose of this study was to develop a feasible technique for bone reparation and further explore the possible applications of Ca-P ceramics in segmental load-bearing bone reparation. HA/TCP ceramics sintered at 1250oC were fabricated into tube-like columns of Φ15mm×30mm with a central canal of Φ4mm. Bone-like apatite was precipitated on the ceramics before implantation. 12 male dogs were used in this study, and a 30mm long segmental bone defect was made in the middle of one femur of each dog. Supported by the fixation of net-cage-structured TC4, the osteoinductive Ca-P ceramic cylinder was used to repair the segmental defect in dog femur. Stress was analyzed by ANSYS. The morphology recovery, function restoration, gait analyses and bone regeneration were evaluated. After implantation at 2, 4 and 8 months, the specimens were harvested respectively. The specimens were evaluated with morphological observation and mechanical testing. Stress analysis showed that the thickness of TC4 net cage was 0.3mm. The morphology recovery of the experimental animal was good and function was restored after 2 months gradually. Aided by stress analysis and by optimizing the design and fixation of implants, Ca-P materials with excellent osteoinductivity could be applied in repairing segmental bone defects.