Surface modification of fiber reinforced polymer composites and their attachment to bone simulating material

The purpose of this study was to investigate the effect of fiber orientation of a fiber-reinforced composite (FRC) made of poly-methyl-methacrylate (PMMA) and E-glass to the surface fabrication process by solvent dissolution. Intention of the dissolution process was to expose the fibers and create a macroporous surface onto the FRC to enhance bone bonding of the material. The effect of dissolution and fiber direction to the bone bonding capability of the FRC material was also tested. Three groups of FRC specimens (n = 18/group) were made of PMMA and E-glass fiber reinforcement: (a) group with continuous fibers parallel to the surface of the specimen, (b) continuous fibers oriented perpendicularly to the surface, (c) randomly oriented short (discontinuous) fibers. Fourth specimen group (n = 18) made of plain PMMA served as controls. The specimens were subjected to a solvent treatment by tetrahydrofuran (THF) of either 5, 15 or 30 min of time (n = 6/time point), and the advancement of the dissolution (front) was measured. The solvent treatment also exposed the fibers and created a surface roughness on to the specimens. The solvent treated specimens were embedded into plaster of Paris to simulate bone bonding by mechanical locking and a pull-out test was undertaken to determine the strength of the attachment. All the FRC specimens dissolved as function of time, as the control group showed no marked dissolution during the study period. The specimens with fibers along the direction of long axis of specimen began to dissolve significantly faster than specimens in other groups, but the test specimens with randomly oriented short fibers showed the greatest depth of dissolution after 30 min. The pull-out test showed that the PMMA specimens with fibers were retained better by the plaster of Paris than specimens without fibers. However, direction of the fibers considerably influenced the force of attachment. The fiber reinforcement increases significantly the dissolution speed, and the orientation of the glass fibers has great effect on the dissolving depth of the polymer matrix of the composite, and thus on the exposure of fibers. The glass fibers exposed by the solvent treatment enhanced effectively the attachment of the specimen to the bone modeling material.     

Comparison of imaging methods used for dental implant osseous integration assessment

Two different imaging techniques used to determine bone tissue response to dental implants were compared. Dental implants were implanted into the maxillae of 18 pigs, which were sacrificed after 4, 8 and 12 weeks. Implants with surrounding bone tissue were retrieved for methyl methacrylate histology and contact radiography. On identical sections peri-implant bone density and bone implant contact (BIC) ratio were assessed with two different imaging methods. Evaluation of Giemsa eosin stained and contact radiographed sections showed direct osseous integration for all implants and both methods showed a strong correlation with correlation coefficient r = 0.930 (P < 0.0001) for peri-implant bone density and r = 0.817 (P < 0.0001) for bone implant contact ratio. While the two imaging methods showed moderate differences for peri-implant bone density there were significant differences between the BIC values determined. In general, contact radiography tends to underestimate BIC for approximately 4.5 % (P = 0.00003).     

Response of primary fibroblasts and osteoblasts to plasma treated polyetheretherketone (PEEK) surfaces

Polyetheretherketone (PEEK) is a synthetic polymer with suitable biomechanical and stable chemical properties, which make it attractive for use as an endoprothetic material and for ligamentous replacement. However, chemical surface inertness does not account for a good interfacial biocompatibility, and PEEK requires a surface modification prior to its application in vivo.In the course of this experimental study we analyzed the influence of plasma treatment of PEEK surfaces on the cell proliferation and differentiation of primary fibroblasts and osteoblasts. Further we examined the possibility of inducing microstructured cell growth on a surface with plasma-induced chemical micropatterning.We were able to demonstrate that the surface treatment of PEEK with a low-temperature plasma has significant effects on the proliferation of fibroblasts. Depending on the surface treatment, the proliferation rate can either be stimulated or suppressed. The behavior of the osteoblasts was examined by evaluating differentiation parameters.By detection of alkaline phosphatase, collagen I, and mineralized extracellular matrix as parameters for osteoblastic differentiation, the examined materials showed results comparable to commercially available polymer cell culture materials such as tissue culture polystyrene (TCPS). Further microstructured cell growth was produced successfully on micropatterned PEEK foils, which could be a future tool for bioartificial systems applying the methods of tissue engineering.These results show that chemically inert materials such as PEEK may be modified specifically through the methods of plasma technology in order to improve biocompatibility.The first two authors share first authorship.     

Mineralization of Collagen‐Coated Electrospun Poly(lactide‐co‐glycolide) Nanofibrous Mesh to Enhance Growth and Differentiation of Osteoblasts and Bone Marrow Mesenchymal Stem Cells

To obtain the biomimetic scaffolding materials for bone tissue engineering, poly(lactide‐co‐glycolide) (PLGA) nanofibrous mesh (NFM) was mineralized in a 5× simulated body fluid (SBF) for different time after it was treated by air plasma for 15 min and subsequent collagen coating. The apatite particles were nucleated on the surface of individual nanofibers, gradually grew up, and finally covered the whole NFM surface. The mineral aggregates were mainly composed of tiny hydroxyapatite (HA) nanoparticles, whose content reached a constant value of 54 µg · cm^?2 after 9 days. The collagen coating and apatite deposition enhanced the NFM strength pronouncedly too. In vitro cell culture demonstrated that the non‐ or less mineralized NFMs were more beneficial of cell spreading and proliferation than those highly mineralized NFMs, but the latter ones could strongly promote secretion of alkaline phosphatase (ALP) by osteoblasts after cultured for 14 days. Moreover, the highly mineralized NFMs also could significantly up‐regulated ALP activity and calcium synthesis of bone marrow mesenchymal stem cells (BMSCs), demonstrating that these NFMs are more favorable of the osteoblast phenotype expression and osteogenic induction. Therefore, the biomimetic apatite deposited PLGA/collagen NFM could be a promising candidate scaffold for bone tissue engineering.     

Development of a surface to increase retinal pigment epithelial cell (ARPE-19) proliferation under reduced serum conditions

Age related macular degeneration of the eye is brought about by damage to the retinal pigment epithelium (RPE) and is a major cause of adult blindness. One potential treatment method is transplantation of RPE cells grown in vitro. Maintaining RPE cell viability and physiological function in vitro is a challenge, and this must also be achieved using materials that can be subsequently used to deliver an intact cell sheet into the eye. In this paper, plasma polymerisation has been used to develop a chemically modified surface for maintaining RPE cells in vitro. Multiwell plates modified with a plasma copolymer of allylamine and octadiene maintained RPE cell growth at a level similar to that of TCPS. However, the addition of bound glycosaminoglycans (GAGs) to the plasma polymerised surface significantly enhanced RPE proliferation. Simply adding GAG to the culture media had no positive effect. It is shown that a combination of plasma polymer and GAG is a promising method for developing suitable surfaces for cell growth and delivery, that can be applied to any substrate material.     

Osteogenic activity of silver-loaded coral hydroxyapatite and its investigation in vivo

In this study, the scaffolds based on mineralized silver-loaded coral hydroxyapatites (SLCHAs) were developed for bone regeneration in the radius of rabbit with a 15-mm infective segmental defect model for the first time. The SLCHAs were achieved by surface adsorption and ion-exchange reaction between Ca²⁺ of coral hydroxyapatite (CHA) and Ag⁺ of silver nitrate with different concentration at room temperature. Release experiment in vitro, X-ray diffraction and scanning electron microscopy equipped with energy-dispersive X-ray spectrometer were applied to exhibit that the scaffold showed some features of natural bone both in main component and hierarchical microstructure. The three-dimensional porous scaffold materials imitate the microstructure of cancellous bone. Mouse embryonic pre-osteoblast cells (MC3T3-E1) were used to investigate the cytocompatibility of SLCHAs, CHA and pure coral. Cell activity were studied with alkaline phosphataseenzyme assay after 2, 4, 6 days of incubation. It was no statistically significant differences in cell activity on the scaffolds of Ag⁺(13.6 μg/mL)/CHA, Ag⁺(1.7 μg/mL)/CHA, CHA and pure coral. The results indicated that the lower silver concentration has little effect on cell activity. In the implantation test, the infective segmental defect repaired with SLCHAs was healed up after 10 weeks after surgery, and the implanted composites were almost substituted by new bone tissue, which were very comparable with the scaffold based on mineralized CHA. It could be concluded that the SLCHAs contained with appropriate silver ionic content could act as biocidal agents and maintain the advantages of mineralized CHA or coral, while avoiding potential bacteria-dangers and toxical heavy-metal reaction. All the above results showed that the SLCHAs with anti-infective would be as a promising scaffold material, which whould be widely applied into the clinical for bone regeneration.     

Promotion of adhesion and proliferation of endothelial progenitor cells on decellularized valves by covalent incorporation of RGD peptide and VEGF

Tissue engineered heart valve is a promising alternative to current heart valve surgery, for its capability of growth, repair, and remodeling. However, extensive development is needed to ensure tissue compatibility, durability and antithrombotic potential. This study aims to investigate the biological effects of multi-signal composite material of polyethyl glycol-cross-linked decellularized valve on adhesion and proliferation of endothelial progenitor cells. Group A to E was decellularized valve leaflets, composite material of polyethyl glycol-cross-linked decellularized valves leaflets, vascular endothelial growth factor-composite materials, Arg-Gly-Asp peptide-composite materials and multi-signal modified materials of polyethyl glycol-cross-linked decellularized valve leaflets, respectively. The endothelial progenitor cells were seeded for each group, cell adhesion and proliferation were detected and neo-endothelium antithrombotic function of the multi-signal composite materials was evaluated. At 2, 4, and 8?h after the seeding, the cell numbers and 3H-TdR incorporation in group D were the highest. At 2, 4, and 8 days after the seeding, the cell numbers and 3H-TdR incorporation were significantly higher in groups C, D, and E compared with groups A and B (P?<?0.05) and cell numbers and the expression of t-PA and eons in the neo-endothelium were quite similar to those in the human umbilical vein endothelial cells at 2, 4, and 8 days after the seeding. The Arg-Gly-Asp- peptides (a sequential peptide composed of arginine (Arg), glycine (Gly) and aspartic acid (Asp)) and VEGF-conjugated onto the composite material of PEG-crosslinked decellularized valve leaflets synergistically promoted the adhesion and proliferation of endothelial progenitor cells on the composite material, which may help in tissue engineering of heart valves.     

The association of hydrogel and biphasic calcium phosphate in the treatment of dehiscence-type peri-implant defects: an experimental study in dogs

Hydrogel polymers have many applications in regenerative medicine. The aim of this study in dogs was to investigate bone regeneration in dehiscence-type peri-implant defects created surgically and treated with (i) biphasic calcium phosphate (BCP) granules alone; (ii) a composite putty hydroxypropyl methylcellulose (HPMC)/BCP (MBCP/putty); and (iii) a polymer crosslinked membrane of silanized-HPMC (Si-HPMC/BCP) compared with empty controls. At 3 months, new bone formation was significantly more important in defects filled with HPMC/BCP or Si-HPMC/BCP compared with spontaneous healing in control (P = 0.032 and P = 0.046 respectively) and more substantial compared with BCP alone. Furthermore, new bone formation in direct contact with the implant surface was observed in all three groups treated with BCP. The addition of HPMC to the BCP granules may have enhanced the initial stability of the material within the blood clot in these large and complex osseous defects. The Si-HPMC hydrogel may also act as an occlusive membrane covering the BCP, which could improve the stability of the granules in the defect area. However, the crosslinking time of the Si-HPMC is too long for easy handling and the mechanical properties remain to be improved. The composite MBCP/putty appears to be a valuable bone-graft material in complex defects in periodontology and implantology. These encouraging results should now be confirmed in clinical studies.     

In vitro blood and fibroblast responses to BisGMA–TEGDMA/bioactive glass composite implants

This in vitro study was designed to evaluate both blood and human gingival fibroblast responses to bisphenol A–glycidyl methacrylate–triethyleneglycol dimethacrylate (BisGMA–TEGDMA)/bioactive glass (BAG) composite, aimed to be used as composite implant abutment surface modifier. Three different types of substrates were investigated: (a) plain polymer (BisGMA 50 wt%–TEGDMA 50 wt%), (b) BAG–composite (50 wt% polymer + 50 wt% fraction of BAG–particles, <50 μm), and (c) plain BAG plates (100 wt% BAG). The blood response, including the blood–clotting ability and platelet adhesion morphology were evaluated. Human gingival fibroblasts were plated and cultured on the experimental substrates for up to 10 days, then the cell proliferation rate was assessed using AlamarBlue assay?. The BAG–composite and plain BAG substrates had a shorter clotting time than plain polymer substrates. Platelet activation and aggregation were most extensive, qualitatively, on BAG–composite. Analysis of the normalized cell proliferation rate on the different surfaces showed some variations throughout the experiment, however, by day 10 the BAG–composite substrate showed the highest (P < 0.001) cell proliferation rate. In conclusion, the presence of exposed BAG–particles enhances fibroblast and blood responses on composite surfaces in vitro.     

Biofilm formation on different materials for tooth restoration: analysis of surface characteristics

The purpose of this study was to evaluate the effect of roughness parameters and hydrophobicity of restorative material used to restore non-carious cervical lesions on the biofilm formation. Four restorative materials were investigated: conventional glass ionomer cement (KF, Ketac Fill Plus, 3M ESPE), resin-modified glass ionomer cement (VT, Vitremer, 3M ESPE), nanofilled resin-modified glass ionomer cement (KN, Ketac Nano, 3M ESPE), and nanofilled resin composite (FZ, Filtek Z350 XT, 3M ESPE). Forty disk specimens were prepared from each material, dived in four groups. Five samples were used for topography parameters analysis using a 3D profilometry. The amplitude parameters (Sa and Sq), spatial parameter (Sds), and hybrid parameter (Ssc) were extracted in area using cut-off of 0.25 mm. Hydrophobicity was determined by the contact angle measurement of deionized water on the surface. The biofilm collected from a 24-year-old subject was grown on modified brain–heart infusion agar under aerobic conditions at 37 °C for 24 h. Each test disk was immersed in 200 µL of biofilm suspension (n = 10) and incubated for 24 h at 37 °C. Biofilm was evaluated after 24 h formation on each disk after stained with 1 % fluorescein using confocal laser-scanning microscopy. Data were analyzed using one-way ANOVA and Tukey test (α = 0.05), Pearson correlation was used to compare topography parameters with biofilm formation. Significant differences were found in related amplitude parameters (Sa and Sq, FZ = KN > VT > KF). KN presented the highest hydrophobicity. FZ and KN presented the lowest thickness and biovolume of biofilm when compared with VT and KF. All topography parameters were significantly correlated with biofilm formation. FZ and KN, material with nanoparticles presented better performance-related topography parameters and biofilm formation. Clinical relevance: The incorporation of nanotechnology into restorative materials promotes better surface topography with lower biofilm formation.     

An exploratory study on the efficacy of rat dedifferentiated fat cells (rDFATs) with a poly lactic-co-glycolic acid/hydroxylapatite (PLGA/HA) composite for bone formation in a rat calvarial defect model

In the last two decades, tissue-engineering approaches using scaffolds, growth factors, and cells, or their combination, have been developed for the regeneration of periodontal tissue and bone. The aim of this study was to examine the effects of rat dedifferentiated fat cells (rDFATs) with a poly lactic-co-glycolic acid/hydroxylapatite (PLGA/HA) composite on bone formation in rat calvarial defects. Twenty animals surgically received two calvarial defects (diameter, 5 mm) bilaterally in each parietal bone. The defects were treated by one of the following procedures: PLGA/HA+osteo-differentiated rDFATs implantation (PLGA/HA+rDFATs (OD)); PLGA/HA+rDFATs implantation (PLGA/HA+rDFATs); PLGA/HA implantation (PLGA/HA); no implantation as a control. The animals were euthanized at 8 weeks after the surgery for histological evaluation. The PLGA/HA composite was remarkably resorbed and the amounts of residual PLGA/HA were very slight at 8 weeks after the surgery. The PLGA/HA-implanted groups (PLGA/HA+rDFATs (OD), PLGA/HA+rDFATs and PLGA/HA) showed recovery of the original volume and contour of the defects. The newly formed bone area was significantly larger in the PLGA/HA group (42.10 ± 9.16 %) compared with the PLGA/HA+rDFATs (21.35 ± 13.49 %) and control (22.17 ± 13.08 %) groups (P < 0.05). The percentage of defect closure (DC) by new bone in the PLGA/HA+rDFATs (OD) group (83.16 ± 13.87 %) was significantly greater than that in the control group (40.61 ± 29.62 %) (P < 0.05). Furthermore, the PLGA/HA+rDFATs (OD) group showed the highest level of DC among all the groups. The present results suggest that the PLGA/HA composite is a promising scaffold and that PLGA/HA+DFATs (OD) may be effective for bone formation.     

Poly(ε-caprolactone)-based membranes with tunable physicochemical,bioactive and osteoinductive properties

In contrast to currently used materials, membranes for the treatment of bone defects should actively promote regeneration of bone tissue beyond their physical barrier function. What is more, both material properties and biological features of membranes should be easily adaptable to meet the needs of particular therapeutic applications. Therefore, the role of preparation methods (non-solvent-induced phase separation and thermal-induced phase separation) of poly(ε-caprolactone)-based membranes and their modification with gel-derived bioactive glass (BG) particles of two different sizes (<45 and <3 μm) in modulating material morphology, polymer matrix crystallinity, surface wettability, kinetics of in vitro bioactivity and also osteoblast response was investigated. Both surfaces of membranes were characterised in terms of their properties. Our results indicated a possibility to modulate microstructure (pore size ranging from submicron to hundreds of micrometres), wettability (from hydrophobic to fully wettable surface) and polymer crystallinity (from 19 to 60%) in a wide range by the use of various preparation methods and different BG particle sizes. Obtained composite membranes showed excellent in vitro hydroxyapatite forming ability after incubation in simulated body fluid. Here we demonstrated that bioactive layer formation on the surface of membranes occurred through ACP–OCP–CDHA–HCA transformation, that mimic in vivo bone biomineralization process. Composite membranes supported human osteoblast proliferation, stimulated cell differentiation and matrix mineralization. We proved that kinetics of bioactivity process and also osteoinductive properties of membranes can be easily modulated with the use of proposed variables. This brings new opportunities to obtain multifunctional membranes for bone regeneration with tunable physicochemical and biological properties.     

Analysis of the factors affecting the inherent viscosity of oriented polylactides during hydrolytic degradation

This study focuses on analyzing the effects of several factors on the rate of decay of inherent viscosity (iv) during hydrolytic degradation. The analysis was made for oriented PLLA, 96L/4D PLA and 80L/20D,L PLA. The analyzed polymers were found to have identical rate of iv loss (P < 0.05), given that the materials have otherwise similar initial material properties. The effect of the post-processing residual monomer was dose dependent, i.e. the higher the monomer content the faster the degradation (P < 0.05). Samples with a smaller diameter (1.1 mm) were found to have a faster rate of iv loss than the samples with a larger diameter (4 mm) (P < 0.05). A multiple linear regression analysis was used to create a five-component linear model to predict changes in the materials’ inherent viscosity. This model yielded accurate predictions during the initial stages of the hydrolytic degradation process where the iv loss was virtually linear.     

Improved angiogenic cell penetration in vitro and in vivo in collagen scaffolds with internal channels

Porous scaffolds are limited in volume due to diffusion constraint and delay of vascular network formation. Channels have the potential to speed up cellular penetration. Their effectiveness in improving angiogenic cell penetration was assessed in vitro and in vivo in 3-D collagen scaffolds. In vitro, channelled and non-channelled scaffolds were seeded with vascular smooth muscle cells. Results demonstrated that the scaffolds supported angiogenic cell ingrowth in culture and the channels improved the depth of cell penetration into the scaffold (P < 0.05). The cells reside mainly around and migrate along the channels. In vivo, channels increased cell migration into the scaffolds (P < 0.05) particularly angiogenic cells (P < 0.05) resulting in a clear branched vascular network of microvessels after 2 weeks in the channelled samples which was not apparent in the non-channelled samples. Channels could aid production of tissue engineered constructs by offering the possibility of rapid blood vessel infiltration into collagen scaffolds.     

Effects of two low-shrinkage composites on dental stem cells (viability, cell damaged or apoptosis and mesenchymal markers expression)

To investigate the effects of new two low-shrinkage composites SDR^® and Venus^®Bulk Fill on the cell viability, cellular damage and expression of mesenchymal markers on dental stem cells. Specimens from two low-shrinkage composites were eluted with culture medium for 24 h. After 24 h of incubation, cytotoxicity of elutes were evaluated by MTT assay; apoptosis was determined using the DNA-specific fluorochrome Hoechst 33342 and the mesenchymal stem cells markers expression was analyzed by immunofluorescence staining. After 24 h of cell exposure to each extract media, dental stem cells expressed MSCs markers. The interaction among the material and cell line was not significantly correlated [F(1,60) = 2.251, P = 0.39], whereas statistically significant differences among cells lines were observed [F(1,60) = 9.157, P = 0.004], being dental pulp stem cells more resistant that periodontal ligament stem cells. Also, we did not find any significant effect between the tested materials [F(1,60) = 0.090, P = 0.765]. Furthermore, a very low proportion of exposed cells showed condensed or fragmented nuclei, typical of apoptotic cells at 24 h. The results suggest that SDR^® and Venus^® Bulk fill and should be considered when selecting an appropriate resin-based dental restorative material.     

Effect of interface modification on PMMA/graphene nanocomposites

Graphene platelets (GnPs) were surface modified with a long-chain surfactant B200, and then compounded with polymethyl methacrylate (PMMA). B200 provided an anchor into GnPs and a bridge into the matrix, thus creating molecular entanglement between matrix and GnPs. The interface modification promoted the dispersion of GnPs, as no aggregates of GnPs were observed on the fracture surface of the modified composites, in sharp contrast with the unmodified composites. Although GnPs formed clusters in the matrix, bilayer graphene was readily observed under TEM in randomly selected regions; it showed high structural integrity under diffraction pattern. The addition of 2.7 vol% m-GnPs produced 32.8 % improvement in the flexural modulus of PMMA as compared to 9.0 % by unmodified GnPs. At 1.1 vol%, the interface-modified composite showed a 19.6 % improvement in the absorption resistance to ethanol, in comparison with 3.8 % for the unmodified composites. The addition of 2.7 vol% m-GnPs improved fracture toughness of PMMA by 79.2 %, while GnPs enhanced it by 23.9 %.     

One‐step method for the preparation of poly(methyl methacrylate) modified titanium‐bioactive glass three‐dimensional scaffolds for bone tissue engineering

A novel three‐dimensional (3D) titanium (Ti)‐doping meso‐macroporous bioactive glasses (BGs)/poly(methyl methacrylate) (PMMA) composite was synthesised using PMMA and EO₂₀ PO₇₀ EO₂₀ (P₁₂₃) as the macroporous and mesoporous templates, respectively. Unlike the usual calcination method, the acid steam technique was used to improve the polycondensation of Ti‐BGs, and then PMMA was partially extracted via chloroform to induce the macroporous structure. Simultaneously, the residual PMMA which remained in the wall enhanced the compressive strength to 2.4 MPa (0.3 MPa for pure BGs). It is a simple and green method to prepare the macro‐mesoporous Ti‐BGs/PMMA. The materials showed the 3D interconnected hierarchical structure (250 and 3.4 nm), making the fast inducing‐hydroxyapatite growth and the controlled drug release. Besides mentioned above, the good antimicrobial property and biocompatible of the scaffold also ensure it is further of clinical use. Herein, the fabricated materials are expected to have potential application on bone tissue regeneration.Inspec keywords: titanium, bone, tissue engineering, glass, materials preparation, biomedical materials, polymers, porous materials, drug delivery systems, nanomedicineOther keywords: poly(methyl methacrylate), PMMA preparation, 3D titanium‐bioactive glass scaffold, bone tissue engineering, titanium‐doping mesomacroporous bioactive glass, bioactive glass‐PMMA composite, macroporous template, mesoporous template, calcination method, acid steam technique, titanium‐bioactive glass polycondensation, macroporous structure, green method, macromesoporous titanium‐bioactive glass‐PMMA, 3D interconnected hierarchical structure, fast inducing‐hydroxyapatite growth, controlled drug release, bone tissue regeneration, Ti     

Evaluation of the osteogenesis and angiogenesis effects of erythropoietin and the efficacy of deproteinized bovine bone/recombinant human erythropoietin scaffold on bone defect repair

Erythropoietin (EPO) could promote the angiogenesis and may also play a role in bone regeneration. This study was conducted to evaluate the osteogenesis and angiogenesis effects of EPO and the efficacy of deproteinized bovine bone/recombinant human EPO scaffold on bone defect repair. Twenty-four healthy adult goats were chosen to build goat defects model and randomly divided into four groups. The goats were treated with DBB/rhEPO scaffolds (group A), porous DBB scaffolds (group B), autogenous cancellous bone graft (group C), and nothing (group D). Animals were evaluated with radiological and histological methods at 4, 8 and 12 weeks after surgery. The grey value of radiographs was used to evaluate the healing of the defects and the outcome revealed that the group A had a better outcome of defect healing compared with group B (P < 0.05). However, the grey values in group A were lower than group C at week 4 and week 8 (P < 0.05), but at week 12 their difference had no statistical significance (P > 0.05). The newly formed bone area was calculated from histological sections and the results demonstrated that the amount of new bone in group A increased significantly compared with that in group B (P < 0.05) but was inferior to that in group C (P > 0.05) at 4, 8, 12 weeks respectively. In addition, the expression of vascular endothelial growth factor (VEGF) by immunohistochemical testing and real-time polymerase chain reaction at 12 weeks in group A was significantly higher than that in group B (P < 0.05), and also better than that in group C at week 4 and week 8 (P < 0.05), but at week 12 their difference had no statistical significance (P > 0.05). Therefore, EPO has significant effects on bone formation and angiogenesis, and has capacity to promote the repair of bone defects. It is worthy of being recommended to further studies.     

Bonding ability evaluation of bone cement on the cortical surface of rabbit’s tibia

A composite bone cement designated G2B1 that contains β tricalcium phosphate particles was developed as a bone substitute for percutaneous transpedicular vertebroplasty. In this study, both G2B1 and commercial PMMA bone cement (CMW1) were implanted into proximal tibiae of rabbits, and their bone-bonding strengths were evaluated at 4, 8, 12 and 16 weeks after implantation. Some of the specimens were evaluated histologically using Giemsa surface staining, contact microradiography (CMR) and scanning electron microscopy (SEM). Histological findings showed that G2B1 contacted bone directly without intervening soft tissue in the specimens at each time point, while there was always a soft tissue layer between CMW1 and bone. The bone-bonding strength of G2B1 was significantly higher than that of CMW1 at each time point, and significantly increased from 4 weeks to 8 and 12 weeks, while it decreased significantly from 12 weeks to 16 weeks. Bone remodeling of the cortex under the cement was observed especially for G2B1 and presumably influenced the bone bonding strength of the cement. The results indicate that G2B1 has bioactivity, and bone bonding strength of bioactive bone cements can be estimated fairly with this experimental model in the short term.     

Fibrin-mediated endothelial cell adhesion to vascular biomaterials resists shear stress due to flow

In vitro endothelial cell (EC) seeding onto biomaterials for blood-contacting applications can improve the blood compatibility of materials. Adhesive proteins adsorbed from serum that is supplemented with the culture medium intercede the initial cell adhesion and subsequent spreading on material surface during culture. Nevertheless, physical and chemical properties of vascular biomaterial surface fluctuate widely between materials resulting in dissimilarity in protein adsorption characteristics. Thus, a variation is expected in cell adhesion, growth and the ability of cell to resist shear stress when tissue engineering on to vascular biomaterials is attempted. This study was carried out with an objective to determine the significance of a matrix coating on cell adhesion and shear stress resistance when cells are cultured on materials such as polytetrafluoroethylene (PTFE, Teflon) and polyethyleneterephthalate (Dacron), ultra high molecular weight polyethylene (UHMWPE) and titanium (Ti), that are used for prosthetic devices. The study illustrates the distinction of EC attachment and proliferation between uncoated and matrix-coated surfaces. The cell attachment and proliferation on uncoated UHMWPE and titanium surfaces were not significantly different from matrix-coated surfaces. However, shear stress resistance of the cells grown on composite coated surfaces appeared superior compared to the cells grown on uncoated surface. On uncoated vascular graft materials, the cell adhesion was not supported by serum alone and proliferation was scanty as compared to matrix-coated surface. Therefore, coating of implant devices with a composite of adhesive proteins and growth factors can improve EC attachment and resistance of the cells to the forces of flow.